Title of Invention	A METHOD FOR THE PREPARATION OF THE S-OR R-ENANTIOMER OF A DIOL HAVING THE FORMULA II
Abstract	The present invention relates to a method for the preparation of the S- or R- enantiomer of diol having the formula (II) or a salt thereof and/or the opposite enantiomer of an acylated diol having the formula (IV) comprising a) subjecting a racemic compound of formula (II) to selective enzymatic acylation using an acylating agent having the formula (III a), (III b) or (III c) or an isocyanate having the formula R1 -':N=C-O or an isothioyanate having the formula R1-N=C=S; to form a mixture of the starting material of formula (II) in either the R- or the S-form and opposite enantiomer of a compound having the formula (IV); or b) subjecting a racemic compound of formula (IV) to selective enzymatic deacylation to form a mixture of deacylated compound of formula (II) in either the R- or the S- form and the acylated starting material of formula (IV) in the form of the opposite enantiomer; optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (II) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof.

Title of Invention

A METHOD FOR THE PREPARATION OF THE S-OR R-ENANTIOMER OF A DIOL HAVING THE FORMULA II

Abstract

The present invention relates to a method for the preparation of the S- or R- enantiomer of diol having the formula (II) or a salt thereof and/or the opposite enantiomer of an acylated diol having the formula (IV) comprising a) subjecting a racemic compound of formula (II) to selective enzymatic acylation using an acylating agent having the formula (III a), (III b) or (III c) or an isocyanate having the formula R1 -':N=C-O or an isothioyanate having the formula R1-N=C=S; to form a mixture of the starting material of formula (II) in either the R- or the S-form and opposite enantiomer of a compound having the formula (IV); or b) subjecting a racemic compound of formula (IV) to selective enzymatic deacylation to form a mixture of deacylated compound of formula (II) in either the R- or the S- form and the acylated starting material of formula (IV) in the form of the opposite enantiomer; optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (II) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof.

Full Text	The present invention relates to a novel method for the preparation of optically active intermediates useful for the preparation of escitalopram involving selective enzymatic acylation or deacylation. Background of the inventioa Citalopram is a well-known antidepressant drug that has now been on the market for some years. It is a selective, centrally acting serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitor, accordingly having antidepressant activities. Citalopram was first disclosed in DE 2,657,013, corresponding to US 4,136,193. This patent pubUcation i.a. outlines a process for preparation of citalopram jfrom the corresponding 5-bromo-derivative by reaction with cuprous cyanide in a suitable solvent and by alkylation of 5-bromo-phtalane. US Patent No 4,943,590 corresponding to EP-B1-347 066 describes two processes for the preparation of escitalopram (S-enantiomer of citalopram). Both processes use the racemic diol having the fomiula as starting material. According to the first process, the diol of formula (I) is reacted with an enantiomerically pure acid derivative, such as (+) or (-)-a-methoxy-a-trifluoromethyl-phenylacetyl chloride to forai a mixture of diastereomeric esters, which are separated by HPLC or firactional crystallization, whereupon the ester with the correct stereochemistry is enantioselectively converted into escitalopram. According to the second process, the diol of formula (11) is separated into the enantiomers by stereoselective crystalhzation with an enantiomerically pure acid such as (+)-di-p-toluoyltartaric acid, whereupon the S-enantiomer of the diol of the formula (A) is enantioselectively converted to escitalopram. Escitalopram has now been developed as an antidepressant. Hence, there is a desire for an improved method for preparation of escitalopram. It has now been found that the S-enantiomer of the diol of formula (I) above as well as acylated derivatives thereof may be prepared by selective enzymatic acylation of the primary hydroxyl group in the racemic diol to obtain S-diol or an acylated derivative thereof with high optical purity and fiirther that the enantiomers obtained may be separated by a series of isolation and purification operations. The invention Accordingly, the present invention relates to a novel process for the preparation of the S- or R-enantiomer of a diol having the formula wherein R is cyano or a group which may be converted to a cyano group, Z is a group -CH2-N(R'R") wherein R'and R" are C1-6-alkyl, or R'and R" are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl group, the dotted Une represents a double or a single bond and Hal is halogen or a salt thereof, and/or the opposite enantiomer of an acylated did having the formula wherein R," Z, the dotted Une and Hal are as defined above, W is O or S, and R^ is -Y-R', wherein R^ is C1-10-alkyl, C2-10-aIi:enyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from CMO-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(CMo-allcyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl; wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt thereof, comprising a) subjecting a racemic compound of formula or R and R together form a chain of 3 to 5 carbon atoms; provided that W and U are not S when X is S; to form a mixture of the starting material of foraiula (II) in either the R- or the S-form and the opposite enantiomer of the acylated diol having the formula wherein R, W, Hal, R", the dotted hne and Z are as defined above; or b) subj ecting a racemic compound of formula wherein R, Z, W, Hal, the dotted line and R3are as defined above; to selective enzymatic deacylation to form a mixture of deacylated compound of formula wherein R, Hal, the dotted line and Z are as defined above in either the R- or the S-fonn and the acylated starting material of formula (TV) in the form of the opposite enantiomer; optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (H) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof. The invention also relates to methods for the separation of mixtures of an enantiomer of fonnula (IV) from the opposite enantiomer of formula (11) and to the R- and S-enantiomers of the compounds of formula (TV) above. Finally, the invention relates to a method for the preparation of escitalopram and racemic citalopram from the enantiomers of a compound of formula (II) obtained by the selective enzymatic acylation or deacylation according to the invention, or the enantiomers of the optically active acyl derivative of formula (IV) obtained by the selective enzymatic acylation or deacylation according to the invention. Detailed description of the invention Wlien used in connection with the compounds of formula (11), (TV) and (V), the terms "enantiomer", "R-enantiomer", "S-enantiomer", "R-form", "S-form", "R-diol" and ' "S-diol" refer to the orientation og the groups around the carbon atom to which the 4-Hal-phenyl group is attached. ine present invention thus relates in one embodiment to a selective enzymatic acylation as above and in another embodiment to selective enzymatic deacylation as above. Selecrive enzymatic acylation means that the'enzymatic acylation is preferentially effective for conversion of one of the enantiomers of a compound of formula (II) preferentially leaving the other enantiomer of the compound of formula (II) unconverted in the reaction mixture. Selective enzymatic deacylation means that the enzymatic deacylation is preferentially effective for conversion of one of the enantiomers of a compoimd of formula (IV), preferentially leaving the other enantiomer of the compound of formula (IV) unconverted in the reaction mixture. The selective acylation according to the invention thus results in a mixture containing preferentially the compound of formula (II) in the S-form and the compound of formula (IV) in the R-form, or it may result in a mixture containing preferentially the compound of formula (E) in the R-form and the compound'of formula (IV) in the S-form. Likewise, the selective enzymatic deacylation may result in a mixture containing preferentially the compoimd of formula (IV) in the S-fonn and the compound of formula (E) in the R-forai, or it may result in a mixture containing preferentially the compound of formula (IV) in the R-form and the compound of formula (H) in the S-form. The composition mixture obtained after acylation or deacylation according to the invention depend on the specific hydrolase used and the conditions under which the reaction is carried out. Characteristic of the enzymatic acylation/deacylation according to the invention is that a considerably larger portion of one enantiomer is converted than of the other. The optical purity of the diol of formula (11) and/or the acylated compound of formula (IV) obtained by the optical resolution method of the present invention is usually at least 90% ee, preferably at least 95% ee, more preferably at least 97% ee and most preferably at least 98% ee. However, lower values for the optical purity are acceptable. The starting material for the enzymatic method of the invention is a racemic diol of formula (II) or a racemic acyl derivative of formula (TV) In a preferred embodiment of the invention R is halogen or cyano, most preferred cyano- In a further preferred embodiment of the invention Hal is fluoro, In a further embodiment of the invention, the dotted line in formula (II) and (IV) is a single bond. In one embodiment Z is dimethylaminomethyl or a group that may be converted to dimethylaminomethyl. In a suitable embodiment Z is dimethylaminomethyl. Most preferred, Hal is fluoro, R is cyano, the dotted line is a single bond and Z is dimethylaminomethyl. The acylatmg agent used for the encymatic acylation according to the invention may suitable be one of the compounds of formula (Dla), (lUb) and (Hie). In another embodiment the acylating agent used according to the invention is any of the compounds of formula (Ula) and (Illb). According to a further embodiment of the invention, the acylating agent used is a compoimd of formula (IHa). According to a still another embodiment of the invention, the acylating agent used is a compound of formula (Hlb). According to another embodiment of the invention the acylating agent is a compound offonnula(inc). When tlie acylating agent is a compound of foraiula (Ilia), U is suitable O. When the acyalting agent is any of the above, W is suitable 0. When the acylating agent is any of the above, X is suitable O. Suitable, the substituents R°, R1and R2in any of the acylating agents (Ilia), (UJb) and (Hie) defined in any of the embodiments above are as follows: R°, R1and R2are independently selected from C1-6-alkyi, C2-6-alkenyl and C1-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(CiHS-aIkyl)amino or R2is a suitable leaving group, such as succinimidyl, HOBt and pfp, or R0 and R1 together form a chain of 3-5 carbon atoms more suitable R0, R1 and R2 are independently selected from C1-4-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, uitro, cyano, C1-6-alkylamino and di-(CM-alkyI)amino or R^ is a suitable leaving group, such as succinimidyl, HOBt and pfp or R° and R^ together forai a chain of 3-5 carbon atoms, more preferred R0, R1 and R2 are independently selected from C1-3-alkyl, C2.3-aUcenyl and C2-3-al1 Suitably, the substituents R° and R1 in acylating agents of formula (IHa) as defined in any of the embodiments above, are as follows: R° and R1 are independently selected from C1-6s-alkyl, C2.6-allcenyl and C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from C].6-aUcoxy, C1-6 alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-allcylamino and di-(C1-6-alkyl)amino, more suitable R0 and R1 are independently selected from CM-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, CM- alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-{CM-alkyl)amino, preferably R^ and R* are independently selected from Cio-alkyl, C2.3-alkenyl and Cao-alkynyl all of which may all optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyauo, Ci.3-alkylamino and di-(Ci.3-alkyl)aniino, more preferred R° and R^ are independently CM-alkyl and most preferred, and most preferred R^ and R^ is Ci.3-alkyl, in particular unbranched Cio-^alkyl, suitable propyl. Suitably, the substituents R^ and R^ in acylating reagents of formula (lUb) as defined in any of the embodiments above, are as follows: R^andR^ are independently selected from C1-6-alkyl, C2-6-alkenyl and Ci-a-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci-6-alkyl)amino or R is another leaving group, such as succinimidyl, HOBt and pfp, more suitable R and R are independently selected from C1-6-alkyl, Ca^-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-allcylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino or R^ is another leaving group, such as succinimidyl, HOBt and pfp, preferably R^ is selected from Ci.3-alkyl, C2.3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1.3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cio-alkylamino and di-(C1-3-alkyl)amino and R^ is CM-alkyl substituted one or more times with halogen, R" is C2^-alkenyl or R^ is another leaving group, such as succinimidyl, HOBt and pfp, more preferred R^ is Ci.3-alkyl, C2.3-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1.3-allcoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di-(Ci.3-all ~\ 1 R^ meaning C1-6-alkyl substituted one or more times with halogen is a suitable leaving group including groups such as 2,2,2-trichloroethyl and 2,2,2-trifluoroethyl, in particular 2,2,2-trifIuoroethyL In a specific embodiment of the invention, the acylating agent of formula (mb) as above is a compound wherein R is vinyl. According to another specific embodiment of the invention, the acylating agent of foraiula (mb) is a compound as above wherein R^ is propyl. This specific embodiment covers a preferred acylating agent of the invention, namely vinyl butyrate. According to a fiuther embodiment of the invention, the acylating agent is a compound of formula (Hie). Suitably, the substituent R^ in the compound of formula (IIIc) as defined in any of the embodiments above, are as follows: R' is Ci.6-alkyl, C2-6-alkenyl and C2^-alkynyI all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, Ci- V is suitable chloro. The acylating agent used according to the invention may also be an isocyanate of formula R^-N=C=0 or an isothiocyanate of the formula R^-N=C=S. Thus, in a further embodiment of the invention, the acylating agent is an isothiocyanate of the formula R^-N=C=0, According to another embodiment of the invention, the acylating agent is an isocyanate of formula R^-N=C=S. Suitably, the substituent R^ in the isocyanate and the isothiocyanate as defined in any of the embodiments above, are as follows: R^ is Ci-6-alkyl, Ca-e-alkenyl or C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected firom Ci-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-alkyl)amino, more suitable R' is Cu-alkyl, Cz^-allcenyl or C1-6i-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-4-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(CM-alkyl)amino, preferebly R* is Ci.3-alkyl, Czo-alkenyl or Cao-allcynyl all of which may optionally be substituted one or more times with C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-aikylamino and di-(C 1-3-alkyl)amino, more preferred R* is C1-3-alkyl, C2-3-alkenyl or C2-3-alkynyl, and more suitable R^ is C1-3-alkyl, in particular unbranched C1-6-alkyl, such as methyl, ethyl or propyl The invention also covers a method for selective en2ymatic deacylation of a racemic compound of formula (IV) as defined above. Suitable, tlae racemic compound of formula (TV) used, is a compound wherein Y is O, orS. According to a further embodiment the racemic compound of formula (TV) used, is a compound wherein Y is O. In still another embodiment of the invention the racemic compound of formula (IV) used is a compound wherein Y is S. In another embodiment of the invention the racemic compound of formula (IV) used is a compound wherein Y is a bond. Suitably, the substituent R' in the racemic compound (IV) as defined m any of the embodiments above, is as foUows: Cio-alkyl, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected fi-om C1-10-alkoxy, Ci.io-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMO-alkylamino and di-(C1-10-aIkyl)amino, more suitable R' is Ci.io-alkyl, C1-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano, preferably R' is C1-10-alkyl, preferably unbranched Ci.io-alkyl and more preferred R' is unbranched C4-io-aIkyl. According to the invention, selective enzymatic acylation is carried out under conditions substantially suppressing hydrolysis. Hydrolysis, which is the reverse reaction of the acylation reaction, takes place if water is present in the reaction system. Thus, selective enzymatic acylation is preferably carried out in a water-free organic solvent or ahnost anhydrous organic solvent ( enzymes normally require the presence of some water to be active). The examples below, illusfrate how addition of water affects conversion. The percentage of water allowed in a particular reaction system, may be determined by a person skilled in the art. The organic solvent which may be used for the acylation reaction, is not particularly important as long as it does not deactivate the enzyme used. Suitable solvents include hydrocarbons such as hexane, heptane, benzene and toluene; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, 1,4-dioxane, tert-butyl mediyl ether and dimethoxyetlaane; ketones such as acetone, diethyl ketone, butanon, and methyl ethyl ketone; esters such as methyl acetate, ethyl acetate, ethyl butyrate, vinyl butyrate and ethyl benzoate; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane; seconday and tertiary alcohols, such as tert-butanol; nitrogen-containing solvents such as dimethylformamide, acetoamide, formamide. acetonitrile and propionitrile; and orotic polar solvents such as dimethylsulfoxide, N-methylpyrrolidone and hexamethylphosphorous triamide. Among them, hydrocarbons such as hexane, heptane, benzene and toluene, ethers such as diethyl ether, diisopropyl ether, 1,4-dioxane and tert-butyl methyl ether and esters such as vinyl butyrate, are preferred. For one enzymes the most preferred solvents may be aromatic hydrocarbons such as benzene or toluene and ethers, most preferred toluene and for another enzyme the most preferred solvents may be ethers such as 1,4-dioxane (see the examples below). The above solvents may be used singly or in a combination of two or more solvents. The concentration of racemic diol of formula (II) and acylating agent should not be too high as a high concentration of reagents in the solvent may lead to non-selective acylation of the racemic diol. Suitable the concentration of racemic diol and acylating reagent is each below 1,0 M, more suitable below 0,5 M, even more suitable below 0,2 M or even more suitable below 0,1 M. A person skilled in the art will be able to determine the optimal concentration of racemic diol and acylating agent. Selective enzymatic deacylation is preferably carried out in water or a mixture of water and an organic solvent, suitable in presence of a buffer. The organic solvent which may be used in the reaction, is not particularly important as long as it does not deactivate the enzyme used. Suitable organic solvents are solvents miscible with water such as alcohols, acetonitrile, DMF, DMSO, dioxane, DME and diglyme. The skilled person will be able to identify other suitable solvents. A person sldlled in the art will be able to determine the optimal concentration of racemic compound of formula (IV) used in the reaction The stereoselectivity of the enzyme used, may be increased by carrying out the acylation or deacylation in presence of an organic acid and /or an organic base. Accordingly, the present invention also relates to a process wherein the enzymatic acylation or the enzymatic deacylation is carried out in presence of an organic base or an organic acid or a mixture thereof In a particular embodiment, the invention relates to a process wherein the enzymatic acylation or enzymatic deacylation is carried out in the presence of an organic acid, suitable an organic carboxyUc acid. In a further embodiment, the enzymatic acylation is carried out in presence of an organic acid, suitable an organic carboxyUc acid. Suitable the above mentioned organic acid is an aromatic carboxyUc acid or an aliphatic carboxyUc acid. As an organic acid which may be used in the reaction, there may be mentioned, alkyl carboxyUc acids, cycIoalkylcarboxyUc acids, cycloalkylalkylcarboxyUc acids, optionally substituted phenyl-alkylcarboxylic acids and optionaUy substituted phenylcarboxyhc acids. Suitable aUphatic carboxyUc acids, are carboxyUc acids such as formic acid, acetic acid, propionic acid, n-butyric acid, iso-butyric acid, 2-ethylbutyric acid, n-valeric acid, iso-valeric acid, pivaUc acid, n-caproic acid, iso-caproic acid, decanoic acid, crotonic acid, paknitic acid, cyclopentanecarboxyUc acid, cyclohexanecarboxyUc acid, phenyl-CM-alkylcaiboxyUc acids such as 3-phenylpropionic acid, 4-phenyIbutyric acid, oxaUc acid, malonic acid and tartaric acid. Suitable aromatic carboxyUc acids, includes acids such as benzoic acid, p-chlorobenzoic acid, p-nitrobenzoic acid, p-methoxybenzoic acid, p-toluic acid, o-toluic acid, m-toluic acid, naphthoic acid, phthaUc acid and terephthaUc acid, saUcyUc acid, hydrocinnamic acid for instance. Thus, according to one embodiment of the invention, the organic acid used to improve stereoseleselectivity of the enzyme is selected from n-propionic acid, iso-propionic acid, n-butyric acid, iso-butyric acid, iso-valeric acid, 2-ethylbutyric acid, crotonic acid, palmitic acid, cyclohexanecarboxyUc acid, pivaUc acid, benzoic acid and;?-toluic acid, saUcylic acid and 3-phenylpropionic acid. According to a further embodiment of the invention, the carboxyUc acid used is pivaUt acid. The amount of the organic acid to be used is not particularly restricted, but the molar ratio relative to a substrate is usually 0.1 to 10, preferably 1.0 to 3.0, and more preferably 1.0 to 2.0. Altematively, a tertiary amine may be used to improve selectivity of the enzyme, either alone or together with any of the above mentioned organic acid. As suitable organic base there may be mentioned, triethyl amine, pyridine, 4-dimethylaminopyridine and pyridine is preferred. Suitable combinations of organic acid and organic base are benzoic acid and pyridine for example. The amount of the tertiary amine to be used is not particularly restricted, but the molar ratio relative to a substrate is usually 0.5 to 3.0, and preferably 0.5 to 2.0. The enzymatic acylation or deacylation according to the invention is carried out using a hydrolase, such as a lipase, an esterase, an acylase or a protease. Thus, according to one embodiment of the invention, enzymatic acylation is performed with a hydrolase, such as a lipase, an esterase, an acylase or a protease. The enzymes useful according to the invention are such enzymes capable of performing R-selective acylation or S-selective acylation of the primary hydroxy group in the racemic compound of formula (II). According to another embodiment of the invention, enzymatic deacylation is performed with a hydrolase, such as a lipase, an esterase, an acylase or a protease. The enzymes useful according to the invention are such enzymes capable of performing R-selective deacylation or S-selective deacylation of the acyl group in the racemic compound of formula (IV). As used herein "hydrolase" and "enzyme" either generally or in relation to a specific enzyme, means not only the enzyme itself, but also cultured products containing the enzyme, such as culture fluid containing a cell body, or a cultured cell body, and processed product of the cultured product (for example a crude extract, a freeze-dried micrcorganism or cell, an acetone dried microorganism or cell, a ground product of such microorganism or cell, or the like). Additionally, the "enzyme" or "hydrolase" may be immobilized as the enzyme itself or as a cell body by loiown techniques, and may be used in immobilized form. The immoDiiizauon may oe camea out by metnods Known to me person siaiied m the art, such methods include, for example carrier bonding, cross linking, encapsulation and the like-Thus, in one embodiment of the invention the hydrolase is used in the form of an immobilized enzyme or Cross-Linked Enzyme Crystal (CLEC) enzymes. It has been found that enzymatic acylation accordmg to the invention may be carried out using Novozyme 435, from Candida antartica, LipoZyme TLIM from Thermomyces lanuginosus or Lipoprotein Lipase pseudomonas sp. (isolated from Pseudomonas Cepacia and obtained from Fluka), and particularly good results have been found when using Novozyme 435, from Candida antartica or Lipoprotein Lipase pseudomonas sp. Thus, according to one embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein Upase, Candida antartica Upase B or Thermomyces lanuginosus lipase. According to another embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein lipase or Candida antartica lipase B. As mentioned above, use of one of the above mentioned enzymes according to the invention also covers the use of cultured products containing the enzyme, such as culture fluid containing a cell body, or a cultured cell body, processed product of the cultured product and any immobilized forms of these enzymes/cultured products. The use of any of the above specifically mentioned enzymes according to the invention also covers the use of mutants, variants or any equivalents of the above specifically mentioned enzymes, which are capable of performing the selective acylation or deacylation according to the invention. The variants or equivalents thereof may be isolated from various strains of Pseudomonas, Candida or Thermomyces, or any other source, or they may be prepared by mutation of the DNA encoding the above mentioned enzymes leading to variations in the amino acid composition of the enzyme. Suitable the mutants or variants of the above mentioned enzymes axe variants and mutants where single amino acids have been removed or replaced by other amino acids, and suitable the amino acid sequence of the variant or mutant is more than 60 % identical, preferably more than 80 % or most preferred more than 90 % identical to the above mentioned enzymes. Thus, according to one embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein Upase or a mutant or variant thereof. Preferably Pseudomonas sp. lipoprotein Upase is used. According to another embodiment of the invention, the enzyme used is Candida antartica lipase B or a mutant or variant thereof. Preferably, the enzyme used is Candida antartica Upase B. According to a further embodiment of the invention the enzyme is Novozyme®435 (Candida antartica Upase B immobiUzed on acrylic resin, available from the company Novozymes A/S). According to still another embodiment of the invention, the enzyme used is Thermomyces lanuginosus Upase or a mutant or variant thereof. Preferably the enzyme used is Thermomyces lanuginosus Upase. According to a further embodiment of the invention, the enzyme used is Lipozyme^^ TLIM, also available from the company Novozymes A/S. The preferred reaction conditions for enzymatic acylation/deacylation differ depending on the particular enzyme used, whether it is immobiUsed or not etc. A suitable temperature for the reaction lies between 0-80 °C, more preferably between 20-60 °C, or more preferred between 30-50 °C. The amount of enzyme to be used is not particularly restricted, but is usually O.Ol-I.O, nrftferahlv 0.02-0.5 and mnrf* nr^^f^araKU/ 0 09-0 1 ^c \x/fiaVit ratin r^lativp tn CTiKctroto The reaction may be carried as a batch process or it may be carried out as a continous process. The enzyme may be used in a plurality of batches repeatedly or continuously. The reaction time is not particularly restricted, and will depend on the enzyme used and the scale and type production method (batch or continues). The present invention also relates to an S- or R-enantiomer of a compound having the foraiula (TV) wherein R, Hal, R"', W, the dotted line and Z are as defined above,or a salt thereof According to one embodiment of the invention, the optically active acyl derivative above is the S-enantiomer. According to another embodiment of the invention, the optically active acyl derivative above is the R-enantiomer. According to one embodiment of the invention the R- or S-enantiomer above is a compound wherein R is halogen or cyano, preferably R is cyano. According to a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Hal is fluoro. According to a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein the dotted line represents a single bond. In another embodiment of the invention, the R- or S-enantiomer above is a compound wherein Z is dimethylaminomethyl or a group that may be converted to dimethylaminomethyi, more suitable Z is dimethylaminomethyl, and the other substituents are as defined above. In another embodiment of the invention, the R- or S-enantiomer above is a compound wherein Z is dimethylaminomethyl, Hal is fluoro, the dotted line represents a single bond, and R is cyano or halogen, suitable cyano. In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is 0, or S, preferably Y is O and the other substituents are as defined above. In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is S and the other substituents are as defined above. In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is a bond and the otlier substituents are as defined above. In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is NH and the odier substituents are as defined above. Suitable, the R- or S-enantiomer as defined in any of the embodiments above is a compound wherein R^ is C1-6-alkyl, C2-6-alkenyl or C2-6-alIcynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(C1-6-alkyl)amino, more suitable R' is CM-alkyl, C1-6-allcenyl or C1-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(CM-alkyl)amino, preferably R^ is C1-3-alkyl, Ca-s-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with substituents sefected from Ci.3-alkoxy,'C1-3-allcylthio, hydroxy, halogen, amino, nitro, cyano, Cio-alkylamino anddi-(Ci,3-alkyl)aniino and most preferred R is Cio-allcyi, in particular imbranched Ci,3-alkyL According to another suitable embodiment of the invention, the R- or S-enantiomer as defined in any of the embodiments above is a compound wherein R^ is as follows: Ci. lo-alkyi, C2-10-aIkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times ^^'ith substituents selected from Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci- After the completion of the acylation reaction or the deacylation reaction, an enantiomer of the diol derivative represented by the formula (II) is obtained as a -mixture with the opposite enantiomer of the compound of formula (IV), This reaction mixture is then optionally separated from the enzyme. In order to obtain the desired enantiomer of formula (11) and/or (IV) with high chemical purity, it is necessary to separate it form the opposite enantiomer of formula (TV) and (II), respectively, in an efQcient manner. However, it is difficult to efficiently separate the enantiomers by generally known methods because the structure of the acyl derivative of formula (TV) is so similar to the stmcture of the diol of formula (n). The present inventors have made intensive investigations and have as a result found that the enantiomer of a compound of formula (H) in the form of a salt with an acid is efficiently distributed into an aqueous layer and the opposite enantiomer of a compound of formula (IV) in the form of a salt with the acid is efficiently distributed into an organic layer when the reaction mixture is treated with a mixed solvent containing organic solvent and water in tlie presence of an acid. Thus, according to another embodiment, the invention relates to a method for the isolation and purification of an acyl derivative having the formula wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line is a double or a single bond and Z is a group -CH2-N(R'R') wherein R'and-R-' C1-6-alkyl, or R'and R" are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethyiaminomethyl group, W is O or S and R^ is -Y-R^ wherein Y is a bond, 0, S or NH and R^ is C1-10-allcyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted with one or more substituents selected from Ci-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci. lo-alkylamino, di-(C1-10-allcyl)ainino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Ci-10-alkyl, C1-10-alkenyl, CJAQ-alkynyl, Ci-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMO-alkylamino and di-(Ci.io-alkyl)amino, or a salt thereof and/or a diol of formula wherein R, Hal, the dotted line and Z are as defined above, or a salt thereof, from a mixture containing a compound of fonnula (IV) and a diol of formula (II), which comprises: a) treating said mixture containing the acyl of formula (IV) and the diol of formula (II) in a mixture of water and an organic solvent in the presence of an acid; b) separating the aqueous phase contaming the diol of formula (E) as a salt of said acid from the organic phase to obtain an organic phase containing the acyl derivative of fonnula (TV) as a salt of said acid; optionally isolating the compound of formula (II) as the base or as a salt thereof and optionally the compound of fonnula (IV) as the base or as a salt thereof According to one embodiment of the invention R is halogen or cyano, preferably cyano in the method for isolation and purification above. According to another embodiment of the invention Hal is fluoro in the method for isolation and purification above. According to a preferred embodiment, the dotted line represents a single bond. In another embodiment of the invention, Z is dimethylaminomethyl or a group that may be converted to a dimethylaminomethyl group. Suitable Z is dimethylaminomethyl. According to a preferred embodiment of the invention Hal is fluoro, Z is dimethylaminomethyl, the dotted line is a single bond and R is cyano or halogen, preferably cyano. In a further embodiment of the invention the compound of formula (TV) in the method for isolation and purification above, is a compound wherein Y is O, or S, preferably Y is O and the other substituents are as defined above. In a fimher embodiment of the invention tlae compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is S. In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is a bond and the other substituents are as defined above. In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is NH and the other substituents are as defined above. In a preferred embodiment of the invention, the compound of formula (TV) in the method for isolation and purification above, is a compound wherein R^ is Ci-6-alkyl, C2-6-alkenyl or C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, Ci.6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-allcyl)amino, more preferred R^ is C1-4-alkyl, C2-4-alkenyl or C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino, preferably R^ is Cj.3-allcyl, Ci-s-alkenyl and Cio-aUcynyl all of which may optionally be substituted one or more times with substituents selected from Ci.3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-a-alkylamino and di-(C1-3"alkyl)amino and most preferred R is Ci.3-alkyl, in particular unbranched C1-3-alkyl. In another preferred embodiment of the invention, the compound of formula (TV) in the method for isolation and purification above, is a compoimd wherein R^ is as follows: C1-10-alkyl, C2-10-alkenyl or C1-10-allcynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, CMQ-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-6-allc/lamino and di-(CMo-alkyl)amino, more suitable R* is C1-10-alkyl, C1-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano, preferably R* is C1-10-aUcyl, preferably unbranched C1-10-alkyl and more preferred R^ is unbranched C4.io-alkyl. By the above method for isolation and purification, a salt of the R- or S-enantiomer of a compound having4he formula (II) with an acid may be selectively extracted into an aqueous layer and a salt of the opposite enantiomer of a compound of formula (TV) with an acid may be selectively separated into an organic layer by treating, in the presence of the acid, a mixture comprising an enantiomer of formula (II) and the opposite enantiomer of formula (TV), reaction solvent and the like with an organic solvent and water, and in presence of an acid. In this case, the salt of the enantiomer of formula (IV) and the salt of the enantiomer of formula (II) may be separated with little loss of the desired enantiomers. According to one embodiment of the above method for isolation and purification, the S-enantiomer of the compound of formula (TV) is isolated from the R-enantiomer of the compound of formula (II). According to another embodiment of the above method for isolation and purification, the S-enantiomer of the diol of foraiula (II) is isolated from the R-enantiomer of the acyl derivatitive of formula (TV). The amount of water to be used is 1:2 to 1:100, preferably 1:5 to 1:50, as aratio between the compound of fomiula (E) and water. In addition, prior to the extraction with water, the reaction solvent may be evaporated to reduce the amount thereof or may be substituted with another organic solvent. The acid which may be used in the above isolation and purification process is not particularly restricted, but for example, there may be mentioned, mineral acids such as hydrochloric acid, sulfuric acid and phosphoric acid; organic acids, in particular carboxyUc acids, represented by aUphatic carboxylic acids such as formic acid, acetic acid, propionic acid, n-butyric acid, iso-butyric acid, n-valeric acid, iso-valeric acid, pivaUc acid, n-caproic acid, iso-caproic acid, cyclopentanecarboxylic acid, cyclohexanecarboxylic acid, oxalic acid, malonic acid and tartaric acid; or aromatic carboxylic acids such as benzoic acid, p-chlorobenzoic acid, p-nitrobenzoic acid, p-methoxybenzoic acid, p-toluic acid, o-toluic acid, m-toluic acid, naphthoic acid, phthalic acid and telephtalic acid. Among them, organic acids are preferred, in particular carboxylic acids such as n-butyrate, iso-valerate, cyclohexanecarboxylic acid, pivalic acid, benzoic acid and o-toluic acid, and particularly preferred is pivalic acid. Needless to say, the above acids may be used singly or in a combination of two or more species. The amount of the acids to be used is not particularly restricted, but the molar ratio of the sum of the compounds of formula (IV) and (II) and acid, is usually 1:1 to 1:5, and preferably 1:1 to 1:3. Tlie acid used in the present isolation and purification step may be the same as the organic acid used in the acylation or deacylation reaction according to the invention or may be a different one. As an organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, hydrocarbons such as hexane, heptane, benzene and toluene; ethers such as diethyl ether, tetrahydrofiiran, 1,4-dioxane, tert-butyl methyl ether and dimethoxyethane; ketones such as acetone, diethyl ketone and methyl ethyl ketone; esters such as methyl acetate, ethyl acetate, ethyl butyrate and ethyl benzoate; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane. Among them, aliphatic hydrocarbons, such as hexane and heptane and aromatic hyderocarbons, such as benzene and toluene are preferred. Most preferred are aromatic hydrocarbons and most preferred is toluene. The above solvents may be used singly or in a combination of two or more solvents. Temperature of the isolation and purification step is preferably at 0 to 80 °C, and more preferably at 10 to 40 ""C and most preferred at 20 to 30 °C Following, separation of the salts of the compounds of formula (EQ and (TV) according to the above method for isolation and purification, minor amounts of the compound of formula (IV) which is partly mixed in an aqueous layer of the salt of the compound of formula (II) with an acid, and the like can be efficiently removed by wasliing the aqueous layer with an organic solvent. The salt of the diol of formula (II) may be used as an aqueous solution of the salt, and if needed, it may be used as a solution in another solvent or a concentrate obtained by concentration, solvent substitution or the like operation. Furthermore, it can be used as a crystal obtained by crystallization or the like operation. However, it is usual that the diol of formula (II) is be used as a free diol form obtained by the following operations: A mixture containing the diol of formula (11) and an organic solvent and/or its concentrate is obtained by treating an aqueous layer with a general base such as sodium hydroxide or potassium hydroxide to control the pH of the aqueous layer to be at least 9, preferably at least 11, then extractmg the free amine form of the diol of formula (II) with an organic solvent, followed by washing and concentrating the extract. The chemical purity of the diol of formula (II) obtained by the series of isolation and purification method is usually at least 95%, preferably at least 97%, more preferably at least 99% and most preferably at least 99.5%. On the other hand, the compound of formula (TV) obtained by the above operations may be washed with an aqueous phase in order to improve chemical purity of the product. The compound of formula (IV) may be obtained as a free amine form by treating the ammonium salt of the compound of foraiula (TV), obtained by the above operations, with a base. The chemical purity of the diol of formula (TV) obtained by the series of isolation and purification method is usually at least 95%, preferably at least 97%, more preferably at least 99% and most preferably at least 99.5%. Tlie optical purity of the product obtained after separation of the enantiomers of formula (II) and (IV) as above, may be improved before fiirther processing. Improvement of the optical purity may be obtained by chromatography as described in WO 03/011278 or by crystallisation of diastereomeric esters or salts with optically active acids as described in US patent No. 4,943,590. According to a further embodiment of the invention, the mixture of compound of formula (11) and compound of formula (TV) which is separated by the above method for isolation and purification has been prepared by the selective acylation and in another embodiment by selective deacylation according to the invention. The invention also relates to another novel method for the separation of the R- or S-diol of formula (11) from the acyl derivative of formula (IV) of the other enantiomer whereby the desired compound may be isolated and purified. According to this other method for isolation and purificationthe acyl derivative having the formula wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line represents a double or a single bond, Z is a group -CH2-N(R'R') wherein R'and R' are Ci-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl group, W is 0 or S; and R^ is -Y-R^ wherin Y is a bond, 0, S or NH and R^ is C1-10-alkyl, C:.io-aIkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(CMo-alkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from CMQ-alkyl, C2-10-aIkenyl, C1-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci.io-alkylaminoand di-(Ci.io-allcyl)amino, or a salt thereof and/or the diol of formula wherein R, Hal, the dotted line and Z are as defined above, or a salt thereof, iso isolated from a mixture containing a compound of formula (IV) and a compound of formula (H), by a method comprising: a) treating said mixture containing the compound of formula (IV) and the diol of formula (II) in a mixture of water, a protic organic solvent and an apolar organic solvent; b) separating the aqueous phase containing the diol of formula (II), firom the organic phase to obtain an organic phase containing the compound of formula (IV); optionally isolating the diol of formula (II) and/or (IV) as a base and optionally conversion of the compound of formula (II) and /or (IV) to a salt thereof. Any of the above isolated phases (aqueous and organic) may additionally be washed one or more times with an organic or an aqueous solvent, respectively to improve the chemical purity of the product. According to one embodiment of the above method for isolation and purification, the S-diol of the compound of foraiula (II) is separated firom the R-enantiomer of the compound of formula (IV). According to another embodiment of the above method for isolation and purification, tlie S-enantiomer of the compound of fomiula (IV) is separated from the R-enantiomer of the compound of formula (II). According to one embodiment of the invention R is halogen or cyano, preferably cyano in the method for isolation and purification above. According to another embodiment of the invention Hal is fluoro in the method for isolation and purification above. According to a preferred embodiment, the dotted line represents a single bond. In another embodiment of the invention, Z is dimethylaminomethyl or a group that may be converted to a dimethylamiaomethyl group. Suitable Z is dimethylaminomethyl. According to a preferred embodiment of the invention Hal is fluoro, Z is dimethylaminomethyl, the dotted line is a stQgle bond and R is cyano or iialogen, preferably cyano. In a further embodiment of the mvention the compound of foraiula (IV) in the method for isolation and purification above, is a compound wherein Y is O, or S, preferably Y is O and tlie other substituents are as defined above. In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound, wherein Yis S. In a further embodiment of the invention the compound of formula (TV) in the method for isolation and purification above, is a compound wherein Y is a bond and the other substituents are as defined above. In a further embodiment of the invention the compound of formula (TV) 'm the method for isolation and purification above, is a compound wherein Y is NH and the other substituents are as defined above. In a preferred embodiment of the invention, the compound of formula (IV) in the method for isolation and purification above, is a compound wherein R^ is C1-6-alkyl, C2-6-alkenyl or Ci-e-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, Ci-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-alkyl)amino, more preferred R^ is Ci.4-alkyl, Cz^-allcenyl or Ci-i-allcynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-all In anotlaer preferred embodiment of the invention, the compound of formula (IV) in the method for isolation and purification above, is a compound wherein R^ is as follows: C1-10-alkyl, C2-10-alkenyl or C2-10-aIkynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, CMO-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.io-alkyl)amino, more suitable R^ is Ci.io-alkyl, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected fi-om hydroxy, halogen, amino, nitro and cyaho, preferably R is C1-10-alkyl, preferably unbranched d-io-alkyl and more preferred R^ is unbranched C4-io-alkyl As an protic organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, alcohols such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanoI and tert-butanol. The above solvents may be used singly or in a combination of two or more species. As an apolar organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, hydrocarbons such 'as hexane, heptane, benzene and toluene; ethers such as diethyl ether, tert-butyl methyl ether and dimethoxyethane; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane. Among them, preferred are hydrocarbons such as hexane, heptane, benzene and toluene, and more preferred is heptane. The above solvents may be used singly or in a combination of two or more species. According to a specific embodiment of the invention, the mixture of the compound of fonnula (TV) and the diol of formula (II) used in the above method for isolation and purification has been prepared by enzymatic acylation according to the invention and in another embodiment by enzymatic deacylation according to the invention. The optical purity of the product obtained after separation of the enantiomers of fonnula (II) and (IV) as above, may have to be improved before further processing. Improvement of the optical purity may be obtained by chromatography as described in WO 03/006449 or by crystallisation of diastereomeric esters or salts with optically active acids as described in US patent No. 4.943.590. The present invention also relates to a process for the preparation of escitalopram having the formula or a phannaceutically acceptable salt thereof comprising preparation of the S-enantiomerofadiolhavingthe formula wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond, Z is a dimethylaminomethyl group or a group which may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof, or the S-enantiomer of an acylated did having the formula wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R^ wherein R^ is Ci.io-alkyI, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom CI-IQ-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(Ci.i{ralkyI)amino, aryl, aryloxy^ aiylthio and heteroaryl, or R* is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2.io-alkenyl, C2.io-alkynyl, Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkyianiino and di-(Ci.io-alkyl)amino and Y is a bond, 0, S or NH or a salt thereof, in one embodiment by the method for selective enzymatic acylation according to the invention as defined in any of the embodiments above audr in another embodiment by the method for selective enzymatic deacylation according to the invention as defined in any of the embodiments above, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and ring closure under basic conditions of the S-enantiomer of formula (Us) or (IVs) or a labile ester derivative thereof to form a compound of foraiula followed by, in either order, if R is not cyano conversion of the group R to a cyano group, if the dotted line represents a double bond reduction to a single bond, if Z is not dimethyiaminometfayl conversion of the group Z to a dimethylaminomethyl group and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolation of escitalopram or a phannaceutically acceptable salt thereof According to one embodiment of the invention, the S-enantiomer of formula (lis) above or the S-eantiomer of formula (TVs) used for the preparation of escitalopram is separated from the R-enantiomer of formula (IV) and (11) respectively, before ringclosure. According to one embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (II) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according one of the above novel methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above novel methods for isolation and purification. According to still another embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (U) and the opposite enantiomer of the compound of formula (IV) obtained by enzymatic deacylarion has been separated from each other by the isolation and purification process according one of the above novel methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above novel methods for isolation and purification. According to another embodiment of the invention, the S-enantiomer of foraiula (lis) above or the S-enantiomer of formula (TVs) used for the preparation of escitalopram is not separated from the R-enantiomer of foraiula (IV) and (IT) respectively, before ringclosure. As mentioned above, the group R means cyano or any other group which may be converted to a cyano group. Groups v^hich may be converted to a cyano group include halogen such as chloro, bromo, iodo or fluoro, preferably chloro or bromo. Other groups which may be converted to cyano include CF3-.(CF2)n-SO2-0-, wherein n is 0-8, -OH, -CHO, -CH2OH, -CH2NH2, -CH2NO2, -CH2CI, -CHiBr, -CH3, -NHR^ -CHNOH, -COOR^ -CONRV wherein R^ is hydrogen or C1-6 alkylcarbonyl, and R^ and R^ are selected from hydrogen, optionally substituted C1-6 atkyl, aryl-C1-6 alkyl or aryl and, a group of fomiula wherein Z is O or S; R^ - R^ are each independently selected from hydrogen and Ci-6 alkyl or R^ and R^ together form a C2.5 alkylene chain tiiereby forming a spiro ring; R^° is selected from hydrogen and C1-6 aUcyl, R^^ is selected from hydrogen, Ci,6 alkyl, a caxboxy group or a precursor group therefore, or R^ and R together forai a C2-5 alkylene chain thereby forming a spire ring. When R is halogen, in particular bromo or chloro, conversion to a cyano may be carried out as described in US 4,136,193, WO 00/13648, WO 00/11926 and WO 01/02383. According to US 4,136,193 conversion of a bromo group to a cyano group, is carried out by reaction with CuCN. wo 00/13648 and WO 00/11926 describe tlie conversion of a halogen or a triflate group to a cyano group by cyanation with a cyanide source in presence of a Pd or Ni catalyst. Other processes for the conversion of a bromo compound to the corresponding cyano derivative involve reaction of the bromo compound with magnesium to form a Grignard reagent, followed by reaction with a formamide to form an aldehyde. The aldehyde is converted to an oxime or a hydrazone which is converted to a cyano group by dehydration and oxidation, respectively. Alternatively, the bromo compound is reacted with magnesium to form a Grignard reagent,, followed by reaction with a compound containing a CN group bound to a leaving group. A detailed description of the above two procedures may be found in WO 01/02383. Compounds wherein the group R is -CHO, may be converted to the corresponding compounds wherein R is cyano by methods analogous to those described in WO 99/00210. Compounds wherein the group R is NHR^^, wherein R^^ is hydrogen or alkylcarbonyl, may be converted to the corresponding compounds wherein the group R is cyano by methods analogous to those described in WO 98/19512. Compoimds wherein R is a -COOR^ group, may be converted to the corresponding :ompound wherein R is cyano by conversion to the amide via the corresponding acid :hloride or an ester thereof followed by dehydration of the amide. WO 01/68632. Alternatively, a compound where R is -COOH may be reacted with chlorosulfonyl isocyanate in order to form the nitrile; cr treated with a dehydrating agent and a sulfonamide as described in WO 00/44738. Tompounds wherein the group R is -CONR^'^R^'^, wherein R^' and R^"^ are selected from hydrogen optionally substituted allcyl, aralkyl or aryl may be converted to the corresponding cyano compound by methods analogous to those described in WO 98/00081 and W098/1951L Compounds wherein the group R is a group of formula (VIE) may be converted to the corresponding cyano compound by methods analogous to those described in WO 00/23431. Compounds wherein R is OH, -CH2OH, -CH2NH2, -CH2NO2, -CH2CI, -CHaBr, -CH3 or any of the groups above, may be converted to the corresponding cyano compounds by methods analogous to those described in WO 01/68632. Racemic compounds of formula (II) may be prepared by the methods described in the above mentioned patents or by the alkylation method described in US patent No. 4.136.193 or the double grignard reaction described in EP 171 943 or by analogous methods. Racemic compounds of formula (TV) may be prepared from racemic compounds of formula (II) by non-selective acylation using anhydrides, esters, carbonates, isocyanates or isothiocyanates as defined by formulas (IQa), (Hlb), (Die), R^-N=C=0 andR^-N=C=S above. In some cases the racemic compound of formula (E) may be available in the fomi of an acid addition salt, such as the sulphate salt, and in this case a free base of the compound of formula (II) may be obtained by treating the salt with a base in a mixture or water and an organic solvent, to transfer the compound of formula (II) into the organic phase. Preferably, R is cyano in the compounds of romula (H), (lis), (Ilr), (IV) (IVs) (IVr) and (V). If R is not cyano, conversion of the group R to a cyano group is suitably carried out after ringclosure to form a compound of formula (V). Preferably, Hal is fluoro in the compounds of romula (H), (lis), (Er), (IV) (IVs) (IVr) md (V).If Hal is not fluoro, conversion of the group Hal to a fluoro is suitably carried Dut after ringclosure to form a compound of formula (V). A procedure for carrying out :his conversion is described in Speciality Chemicals Magazine, April 2003, page 36-58. Z groups which may be converted to dimethylaminomethyl are groups such as -CH2-L, -CH2-NO2, -MgHal, cyano, aldehyde, -CH2-0-Pg, -CHz-NPgjPga, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, -CO-N(CH3)2, -CH(A^R')(A'R^0, -(A'R")(A'R')(A^R'^, -COOR'VCH2-CO-NH2, -CH=CH-R'« or-CONHR", wherein Pg is a protection group for an alcohol group, Pgi and Pg^ are protection groups for an amino group, R^^ and R'^ are independently selected from C1-6 alkyl, C2-6 alkenyl, C1-6 alkynyl and optionally alkyl substituted aryl or aralkyl groups or R^^ and R'^ together form a chain of 2 to 4 carbon atoms, each of R' - R'^ are independently selected from C1-6 alkyl, Cz^ alkenyl, 2i^ alkynyl and optionally C1-6 alkyl substituted aiyl or aryl-C1-6; alkyl, R'^ is hydrogen or methyl and A', A^ and A^ are selected form O and S; L is a leaving group, such as halogen or -O-SO2-A wherein A is C1-6 alkyl, Cz^ alkenyl, C2-6 alkynyl or optionally C1-6 allcyl substituted aryl or aryl-Ci-6 alkyl. In one embodiment of the invention Z is dimethylaminomethyl, -CH2-L, -CH2-NPg,Pg2, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, -CO-N(CH3)2, aldehyde or -COOR'"^ in the compounds of formula (II), (Us), (Ilr), (IV) (IVs) (IVr) and (V). Compounds wherein Z is -CH2-0-Pg may be converted to the correspondiag compounds wherein Z is dimethylaminomethyl by removal of the protection group to form the corresponding alcohol and thereafter conversion of the alcohol group to a feasible leaving group and reaction of the resulting compound with a) dimetlaylamine or a salt thereof, b) methylamine followed by methylation or reductive amination, or c) with an azide followed by reduction to form the corresponduig amine and tlaereafter methylation or reductive amination, as described in WO 01/43525, WO 01/51478 or WO 01/68631 or by analogous methods. Compounds wherein Z is -CH2-L, wherein L is a leaving group, may be converted fo ' a dimethylaminomethyl group in the same manner. Compounds wherein Z is -CO-N(CH3)2 and -CO-NHR'^ wherein R'^ is hydrogen or methyl, may be converted to the corresponding compounds wherein Z is dimethylaminomethyl by reduction of the amide and if a primary or secondary amine . is formed, followed by methylation or reductive amination to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods. Compounds wherein Z is -CH2-NMe(PgO or -CH2-N(Pgl)(Pg2) may be converted to the corresponding compound wherein Z is dimethylaminomethyl by removal of the protection groups, i.e. to obtain compounds wherein Z is -CH2-NH2 or -CHi-NMe and thereafter methylation of the amino group or reductive amination to form a dimethyaminomethyl grot^ as described in WO 01/43525 or WO 01/68631 or by analogous methods. Compounds wherein Z is -CH(A^R^^)(A^R^) may be converted to the corresponding compounds wherein Z is dimethylaminomethyl by deprotection to form an aldehyde (compounds where Z is -CH2-CA^H) followed reductive amination with dimethylamine to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods. Compounds wherein Z is -C(A^R^^)(A^R^^)(A^R^^ may be converted to the corresponding compounds wherein Z is dimethylaminomethyl i) by hydrolysis to fomi a carboxyhc acid or an ester thereof and thereafter reduction to forai an alcohol and thereafter conversion of the alcohol group to a feasible leaving group, followed by replacement of the leaving group with a dimethylamino group as described above, or ii) conversion of the carboxyhc acid or an ester thereof to an amide by reaction with an amine of NH(Me)2, NHaMe, NH3 or a salt thereof foUowed by, in either order, reduction of the amide and if necessary methylation or reductive amination to form a dimethylamino group as described in WO 01/68631 or by analogous methods. Compounds wherem Z is -COOR^^ may be converted to the corresponding compounds wherein Z is dimethylaminomethyl as described^above, starting with the carboxyhc acid ester. Compounds wherein Z is -CH2-CONH2 may be converted to the corresponding compound wherein Z is dimethylaminomethyl by treatment with hypohalide to form a primary amine followed by methylation of the free amino group or reductive amination to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods. Compounds wherein Z is -CH=CHR^^ may be converted to the corresponding compound wherein Z is dimethylaminomethyl by oxidation to fomi an aldehyde which^may be converted to dimethylaminomethyl by reductive amination as described in WO 01/43525 or WO 01/68631 or by analogous methods. Compounds wherein Z is cyano or-CH2-N02 may be converted to the corresponding compound wherein Z is dimethylaminomethyl by reduction followed by methylation or reductive amination of the free of the amino group formed to form dimethylaminomethyl as described in WO 01/68629 or by analogous methods. Compounds wherein Z is -MgHal may be converted to the corresponding compound wherein Z is dimethylaminomethyl by reaction with dimethylaminomethyl alkyl ether of formula (CH3)2NCH20-alkyl as described in WO 01/68629 or by analogous methods. Preferably, Z is dimethylaminomethyl. If Z is not dimethylaminomethyl, conversion of Z to a dimethylaminomethyl group is suitably carried out after ringclosiure. Suitably, Z is dimethylaminomethyl, -CH2-L, -CH2-NPgiPg2, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, aldehyde, -CO-N(CH3)2, -COOR^ when the dotted line represents a bond. Compoirnds of formula (II) or (IV) wherein Z is dimethylaminomethyl, CH2-NPgiPg2 ,-CH2-NMePgi, -CH2-NHCH3, or -CH2-NH2, in particular compounds wherein Z is dimethylaminomethyl, -CH2-NHCH3, or -CH2-NH2, are particularly well suited for the metliod of purification isolation according to the invention, wherein the compounds of formula (II) and (IV) are separated. In the compounds of romula (11), (lis), (Hi), (IV) (TVs) (TVr) and (V), the dotted line is preferably a single bond. Compounds wherein the dotted line represents a double bond may be converted to the corresponding compound wherein the dotted line is a single bond by the methods described in WO 01/68630, Preferably the reduction is carried out after ringclosure. Enantioselective ring-closure of the compounds of formula (IVs) or (lis) to form the compounds of fomiula (V) may suitably be carried out by treatment of a labile ester derivative of the compound with a base such as KOC(CH3)3 and other alkoxides, N^ and other hydrides, triethylamine, ethyldiisopropylamine or pyridine, at low temperatures in an inert organic solvent, such as tctrahydrofuran, toluene, DMSO, DMF, t'butyl methyl ether, dimethoxyethane, dimethoxymethane, dioxane, acetonitrile or dichloromethane. This process is described in US patent No. 4,943,590. Ringclosure of the compound of formula (lis), is suitably carried out by treatment with a base as described above in presence of an agent capable of forming a labile group with the primary alcohol of the diol, such as methanesulfonyloxy, /?- toluenesulfonyloxy, 10-camphorsuIfonyloxy, trifluoroacetyloxy and triiluoromethanesulfonyloxy and halogen. In some cases, it may be advantageous to exchange the -W-R"^ group m the compound of formula (TVs), for a more labile group before ringclosure is carried out. Such labile groups (leaving groups) could typically be a group selected from methanesulfonyloxy, /7-toluenesulfonyloxy, 10-camphorsulfonyloxy, trifluoroacetyloxy and trifluoromethanesulfonyloxy or halogen. Typically, the compound of formula (IVs) is subjected to hydrolysis to form the compound of formula (lis) with aquesous base, such as NaOH, KOH or LiOH in water or alcohol or a mixture thereof and then reacted with an activated leaving group, such as for example mesylchloride or tosylchloride in an organic solvent in the presence of an organic base. The optical purity of the escitalopram product may have to be improved after ringclosure. Improvement of the optical purity may be obtained by chromatography on a chiral stationary phase or by crystallisation of racemic citalopram base or a salt ^ thereof according to the methods described in WO 03/000672. The R-enantiomer of the compounds of formula (E) and (IV) obtained according to the invention may be used to prepare racemic citalopram and escitalopram by ring closure in acidic environment according to the method described in WO 03/000672. Suitable acids for caxiying out acidic ring closure are raiQeraLacid, a carboxylic acid, a sulfonic acid or sulfonic acid derivative, more suitable H2SO4 orH3P04. Thus in another embodiment, the invention relates to a process for the preparation of racemic citalopram and /or escitalopram or a pharmaceutically acceptable salt thereof comprising preparation of the R-enautiomer of a diol having the formula wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents an optinal bond and Z is a dimethylaminomethyl group or a group which may be converted to a dimedaylaminomethyl group and Hal is halogen, or a salt thereof, or the R-enantiomer of an acylated diol having the formula wherein R, Z, the dotted line and Hal is as defined above, W is 0 or S, and R is -Y-R\ wherein R^ is CMo-alkyl, C2-10-alkenyl or C1-10-alkynyi all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy,' halogen, amino, nitro, cyano, Ci.io-alkylamino, di-(CMo-alkyl)amino, aryi, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaiyl groups may optionally be substituted one or more times with substituents selected from Ci.io-alkyl, Cj-io-alkenyl, C2-10-alkynyl, Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMo-alkylamino and'di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt diereof, in one embodiment by the method for selective enzymatic acylation according to the invention as defined in any of the embodiments above and in another embodiment by the method for selective enzymatic deacylation according to the invention as defined in any of the embodiments above, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and ringclosure under acidic conditions of the R-enantiomer of fonnula (Ur) or (TVs) to form a mixture of the compound of formula wherin Hal, Z, R and the dotted line are as defined above, and a minor amount of the corresponding R-enantiomer, followed by, in either order; if R is not cyano conversion of the group R to a cyano group, if the dotted line represents a double bond reduction to a single bond, if Z is not dimethylaminomediyl conversion of the group Z to a dunethylaminomethyl group and if Hal is not fluoro, conversion of Hal * to a fluoro group, followed by isolation of escitalopram and/ or racemic citalopram or a pharmaceutically acceptable salt thereof, by precipitation of racemic citalopram free base or a salt thereof and recovery of escitalopram form the mother liquor of the precipitation. Suitably, the above method for conversion of R-diol to escitalopram, is used for the preparation of escitalopram. The conversion of of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and conversion of Hal to a fluoro group may be carried out as described above. According to one embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (11) and tlie opposite enantiomer of the compound of formula (IV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according t)ne of the above methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above methods for isolation and purification. According to still another raibodiment of the above method for the pr^aration of escitalopram the mixture of the R- or S-CTiantiomer of a compound of foraiula (II) and the opposite enantiomCT of the compound of formula (IV) obrtained by enzymatic deacylation has been separated from each other by the isolation and purification process according one of the above methods for isolation and purification- According to another embodiment, the mixture has been separated by the other of the above methods for isolation and purification. According to another embodiment of the invention, the S-enantiomer of formula (Or) above or the S-enantiomer of fomiula (TVr) used for the preparation of escitalopram is not separated forai the R-enantiomer of formula (IV) and (IT) respectively, before ringclosure. The optical piirity of the escitalopram product may have to be improved after ringclosure. Improvement of the optical purity may be obtained by chromatography on a chiral stationary phase or by crystallisation of racemic citalopram base or a salt thereof according to the methods described in WO 03/000672. As used herein, the term C1-10-alkyl refers to a branched or imbranched alkyl group having from one to ten carbon atoms inclxisive, such as methyl, ethyl, l-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl, 2-methyl-l-propyl, pentyl, hexyl and heptyl. Ci-6-alkyl refers to a branched or unbranched alkyl group having from one to six carbon atoms inclusive, such as methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl, 2-methyH-propyl, pentyl and hexyL CM-alkyl refers to a branched or unbranched alkyl group having from one to four carbon atoms inclusive, such as methyl ethyl, l-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl and 2-methyl-1-propyl. C1-3-alkyl refers to a branched or unbranched alkyl group having from one to three carbon atoms inclusive, such as methyl, ethyl, l-propyl, 2-propyl. Similarly, C1-10-alkenyl and C2-10-alkynyl designate branched or unbranched alkeuyl and alkynyl groups, respectively, having from two to ten carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl. C2^-allcenyI and C2-^-allcynyl designate branched or unbranched alkenyl and alkynyl groups, respectively, having from two to six carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl Cz^^-alkenyl and C2^-alkynyI designate branched or unbranched alkenyl and alkynyl groups, respectively, having from tv^'o to four carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl. C2-3-alkenyl and C2-3-aIkynyl designate branched or unbranched alkenyl and alkynyl groups, respectively, having from two to three carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, ethynyl and propynyl. The terms CMO alkoxy, CMO alkylthio, CMO alkylamino and di-(C1-10-alkyl)ainino etc. designate such groups in which the alkyl group is CMO alkyLas defined above. The terms C\^ alkoxy, CM alkylthio, C1-6 alkylamino and di-(C1-6-alkyl)amino etc. designate such groups in which the alkyl group is Ci-6 alkyl as defined above. The terms CM alkoxy. CM alkylthio. CM alkylamino and di-(CM-alkyI)amino etc. designate such groups in which the alkyl group is CM alkyl as defined above. The terms C1.3 allcoxy, C1-3 alkylthio, C1.3 alkylamino and di-(C]-3"aIkyl)aniino etc. designate such groups in which the alkyl group is C1-3 alkyl as defined above. Halogen means fluoro, chloro, bromo or iodo. The term aryl refers to a mono or bicycUc carbocyclic aromatic group, such as phenyl, or naphthyl, in particular phenyl. Aryloxy, arylthio refers to such a group wherein aryl is as defined above. The term heteroaryl refers to a 5 or 6 membered monocyclic heteroaromatic group or a bicyclic heteroaromatic group. Suitable the heteroaryl group contains 1-3 heteroatoms selected from 0, S and N. R° and R' may together form a chain of 3 to 5 carbon atoms, thus forming an anhydride. When Z is -CHrNCRTl") wherein R'and R" connected to each other to form a cyclic structure including the N-atom to which they are attached, the cyclic structure form groups, such as groups include piperidin, pyrrohdin, moipholinyl and piperazinyl HOBt means hydroxybenzotriazol and pj^ means pentafluorophenoL i Experimentals In the following examples % conversion and optical purity were measured and calculated as described below: HPLC analysis condition (for conversion rate) used for examples 1-28: Column: YMC-Pack ODS-A 0.46 cm I.D. x 25 cm (made by YMC Co., LTD.) Eluent: 25 mM Phosphate buffer / Acetonitrile = 60/40 Flow rate: LOml/min Temperature: 40°C Detector wavelength: 237 nm HPLC analysis condition (for conversion rate) used for examples 29-42: Column: A Lichrospher RP-8 column, 250 x 4 mm (5 ^un particle size) Eluent: Bufifered MeOH/water prepared as follows: 1,1 ml EtsN added to 150 ml water, 10% H3P04(aq) is added to pH=7 and water is added to a total of 200 ml. The mixture is added to 1,8 L MeOH. Temperature; 35^C Flow rate: 1 mL/min Pressure: 16,0 MPa Detection: UV 254 nm 'Injection volume: 10 microL Conversion rate (%) = P/(S+P) x 100, (P: amount of product, S: amount of residual substrate). HPLC analysis condition (for optical purity) used for examples 1-28: Column: Chiralpak AD 0.46 cm I.D. x 25 cm (made by DAICEL CHEMICAL INDUSTRIES, LTD.) Eluent: Heptane / methanol / ethanol / diethylamine = 85 / 7.5 / 7.5 / 0.1 Flow rate: l.Oml/min Temperature: 30°C Detector wavelength: 240 nm Conversion rate (%) = P/(S+P) x 100, (P: amount of product, S: amount of residual substrate) Super critical fluid chromatogeaphy. Analysis condition (for optical purity) used for examples 29-42: Column: Daicel AD column with the dimensions 250 x 4,6 mm (5 \|j.m particle size) Mobile phase: Carbon dioxide Modifier: Methanol with diethylamine (0.5%) and trifluoroacetic acid (0.5%). Modifier gradient: 1-2% in 4 minutes 2-4% in 4 minutes 4-8% in 4 minutes 8-16%in4minutes 16-32% in 4 minutes 32-45% in 1,62 minutes Temperature: Ambient temperature Flow rate: 2 mL/min Pressure: 20 mPa Detection: UV 230 nm and 254 nm Injection volume: lOmicroL Optical purity (% ee) = (A-B) / (A+B) x 100, (A and B represent corresponding stereo isomer, A>B) E-value - hi ((1-c/lOO) x (1-Es/lOO)) / In ((1-c/lOO) x (l+Es/lOO)) (c: convertion ratio, Es: optical purity of residual substrate) Example 1 (S)-4-[4-Dimethylainino-l-(4'-fluorophenyl)-l'hydroxybutyl1-3-hydroxymethyl-benzonitrile Water (200.7 g) and toluene (440.8 g) was added to (±)-4-[4-dimethylamino-l-(4- fluorophenyl>l-hydroxybutyl]-3-hydroxymethylben2omtrile, 1/2 sulfuric acid salt (100.4 g, 0.257 mol), then stirred for 15 minutes and the temperature thereof was raised up to 60*C. Subsequently, 30% NaOH (34.5 g, 0,259 mol) was added dropwise over 5 minutes until pH of aqueous layer reached 11.4, and stirred for 30 minutes. After stinring, the mixture was allowed to stand for 5 minutes, and separated. The aqueous layer was discarded, and thus-obtained organic layer was washed with 220,0 g of water, and further, concentrated under reduced pressi3xe at 60°C to remove away the remaining moisture. The concentration was adjusted by addition of toluene, to thereby obtain 583.1 g of a solution of 4-[4-dimethylamino-l-(4'-fluorophenyl)- l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene (content of pure 4-[4- dimethylamino-l-(4'--fluorophenyl)-l-hydroxybutyi]-3-hydiDxymethylbenzonitrile: 87.6 g, yield: 99.7%, 0.256 mol). Then, 52.5 g of toluene was poured onto 17.5 g of an immobilized enzyme (Novozym 435), then 583.1 g of the solution of 4-[4- dimethylamino-1 -(4'-fluorophenyI)-1 -hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene was added, and the temperature of the mixture was controlled to 40°C. Subsequently, a mixed solution containing pivalic acid (28.7 g, 0.281 mol), vinyl butyrate (29.2 gm 0,256 mol) and toluene (28.8 g) was added into the above mixture over 20 minutes, and the mixture was stirred at 40°C for 12 hours under a slight flow of nitrogen. After cooling the mixture over 2 hours so as to be 20°C, the mixture was stirred for 1 hour at the same temperature, and stirring was stopped. The enzyme was filtrated, and then the enzyme was washed twice with 105 g of toluene, and these • - • - toluene layers were combined. The toluene layer contained 583.1 g of a solution of 4- [4-dimethylamino-l-(4'-.fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtriIe in toluene (content of pure (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l- hydroxybutyl]-3-hydroxymethyIbenzonitrile: 36.0 g, yield through the enzymatic resolution process: 41.1%, 0.105 mol). A mixed solution containii^ the supernatant and washing fluid was extracted three times wifli water (438 g, 263 g x 2), and 1005.4 g of the obtained aqueous layer was washed with 79 g of toluene for five times. As a result, it was obtained 985,2 g of an aqueous solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophaiyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt {content of pure (S)-4-[4-dimethyfaniino-l-(4'-fluorophenyI)-l-hydroxybutyl]-3-hydroxymethyn3enzoni 32,6 g, yield (through the process from the extraction with water to washing with tohiene): 90.6 %, 0.0952 mol}. To thus-obtained 985.2 g of an aqueous solution of (S)-4-[4-dimethylamino-l-{4-fliiorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was added 218^ g of toluene. Subsequently, 30% NaOH (13.3 g, 0.0998 mol) was slowly added to the mixture with stirring, until pH of aqueous layer reached 11.9, and the mixture was kept stirred for 30 minutes. After stilting, the mixture was allowed to stand 30 minutes and the toluene layer was separated, and the remaining aqueous layer was re-extracted with 142.5 g of toluene. The combined toluene layer was washed twice with 169.0 g of water, concentrated at 60jC under reduced pressure, to thereby obtain 64.0 g of a solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzomtrile in toluene. Said toluene solution contained 32.0 g (0.0935 mol) of (S)-4.[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile. Overall yield was 36.4%. The optical purity determined by HPLC was 98.7 % ee, and chemical purity was 99.9 area %, {(R)-5-cyano-2-[dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 0.04 area %. In the above system, die above "chemical purity" is represented by the following equation. (Chemical purity) = [Area value of 4-[4-dimethylainino-l-(4'-fluorophenyl)-l- hydroxybutyl]-3-hydroxymethylben2onitrile] / [(Area value of all detected compounds) - (Area value of toluene) - (Area value derived from the system)] x 100 (area %) Example 2 A test tube equipped with a stopper was charged witti 50 mg ( 0.146 m mol) of (±)-4-^ [4-dimethylaimno-H4'-fIuorophenyl)-l-hydroxybutyl]-3-hyd^^ 25 mg of various kinds of Upases, 33 mg ( 0.29 m mol) of vinyl butyrate and 1 ml of toluene, and stirred at 40C for 16 hours. For the substrate with the optical purity of at least 50% ee, E-values were calculated. The results are shown in Table 1. Example 3 A test tube eqiiipped with a stopper was charged with 10 to 100 mg (0.029 to 0.29 m mol) of (±)-4-[4-dimethyIamino-1 -{4'-fluorophenyl)-1 -hydroxybutyl]-3" hydroxymethylbenzonitrile, 10 to 100 mg of Novozym 435 (product of Novozymes, E/S ratio = 1.0), 33 to 333 mg (10 eq.) of vinyl butyrate and 1 ml of toluene, and stirred at 30°C for 16 hours. The results are shown in Table 2. Example 4 A test tube equipped with a stopper was charged with 100 mg (0.292 m mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl)-3-hydroxymethyl- benzonitrile, 50 mg of Novozym 435 product of Novozymes), 333 mg ( 2.92 m mol) of vinyl butyrate, 0.292 m mol of various kinds of additives and 1 ml of toluene, and stirred at the temperature of 40®C for 16 hours. The results are shown in Table 3. Example 5 In a 200-ml four-necked flask equipped with a stirrer and a thermometer, 1.5 g (4.38 m mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.45 g of Novozym 435 (product of Novozymes), 0.347 g ( 4.38 m mol) of pyridine, 4.38 m mol of various kinds of acids and 1.00 g (8.76 m mol) of vinyl butyrate were added, and stirred at 40'C for between 16 and 21.5 hours. The result is shown in Table 4. Example 6 In a 200-ml four-necked flask equipped with a stirrer and a thermometer, 3.0 g (8.76 m mol) of (±>4-[4-dimethylaraino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.9 g of Novozym 435 (product of Novozymes), 0.693 g ( 8.76 m mol) of pyridine, 8.76 m mol of various kinds of acids and 2.00 g (17.52 m mol) of vinyl butyrate were added, and stirred at 40^C for between 15.5 and 16.5 hours. The results are shown in Table 5. Example 7 In a 200-inl four-necked flask equipped with a stiirer and a thennometer, 4,5 g (13.1 m mol) of (±)-4-[4-dimethylammo-l-(4'-fluorophenyl)-l-liydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.90 g of Novozym 435 (product of Novozymes), 2.68 g (26.2 m mol) of pivalic acid and 0.900 to 1.500 g (0.6 to 1.0 eq.) of vinyl butyrate were added, and stirred at 40T for 24 hours. The restilts are shown in Table 6. Example 8 In a 500-inl separable flask equipped with a stirrer and a thermometer, 20.0 g (58.4 mmol) of (±)-4-(4-dimethylamino-l-(4'-fluorophenyl)-l-hyd[roxybutyl)-3-hydroxymethylbenzonitrile was dissolved in 144 ml of toluene, then pivalic acid (1.1 to 3 eq.), 4.0 g of Novozym 435 (product of Novozymes) and 6.67 g (58,4 m mol) of vinyl butyrate were added, and stirred at 40'C for 20 to 24 hours in a flow of nitrogen (5 ml/min). The results are shown in Table 7. Example 9 A test tube equipped with a stopper was charged with 10 mg ( 0.029 m mol) of (i)-4- [4-dimethylainino-l-{4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbe^^ , 10 mg of Novozym 435 (product of Novozymes), 0.29 m mol of various kinds of acyl donor and 1 ml of diisopropyl ether, and stirred at 30*C for 16 hours. For the substrate with the optical purity of at least 30% ee, E-values were calculated. The results are shown in Table 8. Example 10 A test tube equipped with a stopper was charged with 10 mg (0.029 m mol) of (i)-4-[4-dimethylaminO' 1 -(4'-fluorophenyl)-1 -hydroxybutyl]-3 -hydroxymethylbeozonitrile, iKj nxg oi iNovozym 4i3 (product of Novozymes), 33 mg ( 0^9 m mol) of vinyl butyrate and 1 ml of various kinds of solvents, and stirred at 30°C for 16 hours. After the reaction, converted ratios and optical purities were analyzed and E-values were calculated. The results are shown in Table 9. Example 11 A 200-nil four-necked flask equipped with stirrer and thermometer was charged with 0.9 to 1.35 g (E/S ratios are 0.2 to 0.3) of Novozym 435 (product of Novozym). A solution of 4.5 g (13.1 mmol) of (±)-4-[4-dimethyIamino-l-(4'-fluorophenyI)-l-hydroxybutyI]-3-hydroxymethylben2onitrile in 30 ml of toluene, 2.01 g of pivaUc acid (19.7 m mol), and 1.50 g (13.1 m mol) of vinyl butyrate was added into the flask. and stirred at between 35 and 45'C for 21 hours. After the reaction, reaction fluid was separated by decantation, and the remaining enzyme was washed with 30 ml of toluene and recovered. The same operation as described above except for using tiie recovered enzyme was repeated four times to cany out the recycle reaction Enzyme activities were calculated according to the following equation. The results are shown in Table 1 a Enzyme activity (U/g) = (Product amount produced,per minute (fimol'min)) / (Enzyme weight (g)) Example 12 (S)-4-[4-DimethylaminO'l-(4'-fluorophenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt A four-necked flask equipped with stirrer and thermometer was charged with 66.6 g of a solution of 10,0 g (0.029 mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and the temperature thereof was controlled so as to be 40^0. Then, 2.27 g (0.029 mol) of pyridine, 2.98 g (0.029 mol) of pivalic acid, 3.35 g (0.029 mol) of vinyl butyrate, and 2.0 g of an immobilized enz3ane(Novozym 435) was added into the above mixture. The reaction mixture was . stirred at 40°CfoT 15 hours under a sUght flow of nitrogen and stirring was stopped. The enzjnne was filtrated from the reaction mixture with Kiriyama funnel, and the enzyme was washed with 21.6 g of toluene. As a result, it was obtained (S)- 4-[4- dimethylamino-l-(4'-fluorophenyl)-l-hydioxybutyl]-3-hydroxy^ (3,6 g, O.OI 1 mol, yield: 36.0%, optical purity: 98.5 %ee), pivalic acid salt The combined toluene layer was washed three times with water (50 ml, 30 ml, 30 ml) at 2(fC, and an aqueous layer containing (S)"4-[4-dimethyiamino-l-(4'-flnorophenyl)- l-hydroxybutyl]-3-hydroxymethyIbenzonitrile (3.5 g, 0.011 mol, extraction yield: 95%), pivalic acid salt was obtained. Example 13 (S>4"[4-Dimethylamino--l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyl-beazonitrile pivalic acid salt » • ... A foxnr-necked flask equipped with stirrer and thermometer was charged with 66.7 g of a solution of 20.0 g (0.058 mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l" hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and the temperature thereof was controlled so as to be 40°C. Then, 5.93 g (0.058 mol) of pivalic acid, 6.62 g (0.058 mol) of vinyl butyrate, and 4.1 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40'C for 18 hours under a slight flow of nitrogen and stirring was stopped The enzyme was filtrated from the reaction mixture with Kiriyama funnel, and the enzyme was washed with 41.3 g of toluene. As a result, 6.7 g (0,020 mol) of (S)-4-[4-dimethylamino-l- (4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was obtained iu the toluene solution, with a yield of 33.5% and an optical purity of 99.7 %ee. Example 14 (S)-4-[4-Dimethylamino-l- (4'-fluorophenyl)'l ■hydroxybutyll-3-hydroxymetfayl'• benzonitrile cyclohexanecarfaoxylic acid salt A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 25.0 g (0.073 mol) of (±)-4-[4-dimethyIamino-l-(4•-fluorophenyI)-l-hydroxybutyl]-3-hyd^oxymethylbenzomtrile in toluene, and the temperature thereof was controlled so as to be 40°C. Then, 5.77 g (0.073 mol) of pyridine, 8.33 g (0.073 mol) of vinyl butyrate, 9.52 g (0.073 mol) of cyclohexanecarboxylic acid, and 5.0 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40'C for 17.5 hours imder a slight flow of mtrogen and stirring was stopped. The enzyme was filtrated, and washed with 54.1 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile (8.6 g, 0.025 mol, yield: 34.4%, optical purity: 98.7% ee), cyclohexanecarboxylic acid salt was obtained, The combined toluene layer was washed five times with water (187.5 ml x 5) at 20°C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-{4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (7.6 g, 0.0221 mol, extraction yield: 89%), cyclohexanecarboxylic acid salt was obtained. Example 15 (S)-4-[4-D™gthylamino-l- (4'fluorophenyl)-l -hydroxybutyll-3-hydroxymetfayl-benzonitrile o-toluic acid salt A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 25.0 g (0.073 mol) of (±)-4-[4-dimethylamino-l-(4-fluorophenyl)-l-hydroxybutyl]-3-hydroxyraethylbenzonitrile in toluene, and the temperature thereof was controlled so as to be 40^C. Then, 5.77 g (0.073 mol) of pyridine, 8.33 g (0.073 mol) of vinyl butyrate, 9.93 g (0.073 mol) of o-toluic acid, and 5.0 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40°C for 21 hours under a slight flow of nitrogen, and then stirring was stopped The enzyme was filtrated firom the reaction mixture with Kiriyama funnel, and washed with 54.1 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (9.7 g, 0.028 mol, yield: 38.8%, optical purity: 97.8% ee), o-toluic acid salt was obtained. The combined toluene layer was washed three times with water (250 ml, 63 ml, 63 ml) at eO^'C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (7.8 g, 0.0228 mol, extraction yield: 80%), o-toluic acid salt was obtained. Example 16 (S)-4-f4-Dimethylaniino-l-(4'-fluoropbenyl)-l-hydroxybutyI1-3-hydroxymethyl-benzonitrile isobutyric acid salt A four-necked flask equipped with stirrer and thermometer was charged with 33.3 g of a solution of 10.0 g (0.029 mol) of (i)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and die temperature thereof was controlled so as to be 40T. Then, 2.29 g (0.029 mol) of pyridine, 2.57 g (0.029 mol) of iso-butyric acid, 3.33 g (0.029 mol) of vinyl butyrate, and 2.0 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40C for 24 hours under a slight flow of nitrogen and stirring was stopped. The enzyme was filtrated off firom the reaction mixture with Kiriyama funnel, and washed with 2L6 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylamino-1 -(4'-fluoropheny 1)-1 -hydroxybutyl] -. 3-hydroxymethylbenzonitrile (3.8 g, 0.011 mol, yield: 38.0%, optical purity: 95.9% ee), isobutyric acid salt was obtained. Example 17 (S)-4-[4-Dimethylamino-l- (4'-fluorophenyl)-l -hydroxybutyl]-3-hydroxymethyl-benzonitrile benzoic acid salt A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 50.0 g (0.146 mol) of (i)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile in toluene, and the temperature tiiereof was controlled so as to be 40°C. Then, 11.5 g (0.146 mol) of pyridine, 33.2 g (0.291 mol) of vinyl butyrate, 17.8 g (0.146 mol) of benzoic acid, and 10.0 g of an irmnobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40°C for 20 hours under a sUght flow of nitrogen, and then stirring was stopped. The enzyme was filtrated firom the reaction mixture with Kiriyama fiinnel, and washed with 108 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyI]- 3-hydroxymethylbenzonitrile (22.0 g, 0.064 mol, yield: 44.0%, optical purity: 99.0% ee), benzoic acid salt was obtained. The combined toluene layer was washed three times with water (500 ml x 3) at eO'^C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4-fluorophenyl)-!-hydroxybutyl]"3-hydroxymethyibenzonitrile (21,0 g, 0.0614 mol, extraction yield: 96yo), benzoic acid salt was obtained. The following examples 18 to 28 illustrate the separation of S-diol from the reaction mixture. Example 18 A test tube equipped with a stopper was charged with 4.3 mg (0,0126 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (compound A), 6.8 mg (0.0165 mmol) of (R)-5-cyano-2-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-benzyl butyrate (compound B), 1 ml of toluene, 1 ml of water, and 0.291 m mol of various kinds of acids (in the case of a dibasic acid, 0.146 m mol is used), and stirred at 40°C for 1 hour. Thus-obtained mixed solvent was separated into aqueous layer and toluene layer, and then the concentrations of compound A and B in each layer were measured. Partition coefficients Ka or Kb are calculated according to the following equation. The results are shown in Table 11. Ka = (Concentration of the compound A in aqueous layer) / (Concentration of the compound A in toluene layer) Kb = (Concentration of the compound B in aqueous layer) / (Concentration of the compound B in toluene layer) Example 19 (S>4-r4-Diniethylagiino-l-(4'-fluorophenylVl-hydroxybmylV3-hydroxy^ benzonitrile pivalic acid salt The enzyme was filtrated fi"om the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 1174.3 g of a toluene solution containing (S)- 4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (44.8 g, 0.131 mol), pivalic acid salt was added 600.3 g of water and extracted. Further twice extraction with 359.7 g of water gave 1375.9 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hyd^oxymethylben2;onitrile, pivahc acid salt. The aqueous solution contained 43.8 g (0.128 mol, yield: 97.8%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyI]-3-hydroxymethylbenzonitrile. Optical purity: 98.4%ee, (R)-5-cyano-2-[dimethyIamino-(4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 10,7 area%. Example 20 (S)-4-[4-Dimethylamino-l - (4'-fluorophenyI)-1 -hydroxybutyl]-3-hydroxymethyl-benzonitrile pivalic acid salt To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3"hydroxymethylbenzonitrile pivalic acid salt (content of (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzonitiile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.018 mol) of ammonium acetate. Then LO g of toluene was added therein and the mixture was stiired for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (0.389 g, 1.136 mmol, yield: 97.3%), pivalic acid salt was obtained By HPLC analysis, it was found that chemical purity was 97.3 area % and (R)-5-cyano-2-[dimethylamino- (4'-fIuorophenyI)-hydroxybutyl] benzyl butyrate: 1.5 area % was contained. Example 21 (SM-r4-Dimethylamino-l-(4'-fluorophenyIVl-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt To 12.6 g of an aqueous solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethyIamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hydroxymethylbCT2X)nitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.013 mol) of lithium sulfate. Then 1,0 g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile, pivalic acid salt (0.394 g, 1.151 mmol, yield: 98.5%) was obtained. By HPLC analysis, it was found that chemical purity was 93.7 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 5.5 area % was contained. Example 22 (S>-4-[4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.011 mol) of ammonium sulfate. Then 1.0 g of toluene was added therein and the mixture was stirred for 10 miiiutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt (0.397 g, 1.159 mmol, yield: 99.3%) was obtained. By HPLC analysis, it was found that chemical purity was 94.7 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluoiophenyl)-hydroxybutyl] ben2yl butyrate: 4.5 area%, was contained Example 23 (S)-4"r4-DimethylaminO'l-(4'-fIuorophenylVl-hydroxyb\ityl]-3-hydroxymethyl-Benzonitrile pivalic acid salt To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethylamino-1 -(4'-£luorophenyI)-1 -hydroxybutyl]-3-hydroxymethylbenzonitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the mefliod of Example 19, was added with L4 g (0.0099 mol) of sodium sulfate. Then LO g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing pivalic acid salt of (S)-4-[4-dimethylamino-l-(4-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbeiizonitrile (0.391 g, 1.142 mmol, yield: 97.8%) was obtained By HPLC analysis, it was found that chemical purity was 94.5 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 4.7 area % was contained. Example 24 (S)-4'C4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroKybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt To 12.6 g of an aqueous solution of (S)-4-[4-dimethyiainino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile pivalic acid salt (content of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 0.7 g (0.012 mol)of sodium chloride. Then 1.0 g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing pivalic acid salt of (S)-4-[4-dimethyIamino-1^4'-fluorophenyl>l-hydroxybutyl]- 3-hydroxymethylbenzomtriIe (0.397 g, 1.159 mmol, yield 99.3%) was obtained. By HPLC analysis, it was found that chemical purity was 93.3 area % and (R)-5-cyano-2-[dimethylamino- (4'-fluorophenyI)-hydroxybutyI] benzyl butyrate: 5.8 area % was contained. Example 20-24 compared to example 19 show that the separation improves by the addition of salts. Example 25 (S)-4-[4-Dimethylaniino-l-(4'fluorophenylVl-hydroxybutyll-3-hydroxymethyl-benzonitrile pivahc acid salt The enzyme was filtrated firom the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.92 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid was adjusted so as to be 40 °C and 13 mL of water was added to the solution and extracted. The solution was further extracted twice with 7.5 mL of water, to obtain a solution of (S)-4-[4-dimethyIaniino-l-(4'-£Iuorophenyl)-l-hydroxybutyl]-3-hydroxymetfaylbenzonitrile, pivalic acid salt. In the above aqueous solution, 0.886 g (2.588 nmiol, yield: 88.6%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3'hydroxymethylbenzonitrile was contained. Example 26 (S)-4-[4"Dimethylamino-l-(4^fluorophenyl)-l"hydroxybutyl1'3-hydroxymethyl-benzonitrile pivalic acid salt The enzyme was filtrated firom the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.920 mmol) of (S)-4-[4-dimethylamino-l-(4'- f f[uorophenyl)-l-hydroxybutyl]-3-hydroxymethylbOT2om1iile, pivahc acid was adjusted so as to be 40°C and 11.6 mL of water was added to the solution and extracted. The solution was extracted further three times with 5.8 mL of water to obtain a solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt. In the above aqueous solution, 0.879 g (2.567 mmol, yield 87.9%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)4-hydroxybutyl]-3-hydroxymethyibenzonitrile was contained. Example 27 (S)-4-[4-Dimethylamino-1 - (4' -fluorophenyi)-1 -hydroxybutyl]-3 -hydroxymethyl-benzonitrile pivalic acid salt The enzyme was filtrated fi-om the enzyme reaction mixture obtaiaed by the method according to Example 1, and washed twice with toluene. Thus-obtained 10.4 g of toluene solution containing 1.0 g (2.92 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivahc acid was added 13.0 mL of water and extracted. The solution was extracted further twice with 7.5 mL of water to obtain an aqueous solution of (S)■4-[4-dimethylamino-l-(4'-fluorophenyl)-I-hydroxybutyI]-3-ydroxymethylbenzonitriIe, pivalic acid salt. In the above aqueous solution, 0.954 g (2.786 mmol, yield 95.4%) of (S)-4-[4-dirQethylaniiQO-l-(4-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was contained. Example 28 (S)-4-[4-Dimethylamino-l-(4^-fluorophenyl)-l-hydroxybutyl>3-hydroxymethyl-benzonitrile pivahc acid salt The enzyme was filtrated from the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.90 mmol) of (S)-4-[4-dimethylamino-l- (4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was added with 0.43 g (4.210 mmol) of pivalic acid and the mixture was extracted with 13.0 mL of water. The solution was fiirther extracted twice with 7.5 mL of water to obtain an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivaUc acid salt In the above aqueous solution, 0.947 g (2.766 mmol, yield: 94.7%) of (S)-4-[4-dimethylamino-l-(4'-fluo^ophenyl)-l-hydroxybutyI]-3-hydroxymethylbe^zonit^ile was contained. A comparison of example 25, 26, 27 and 28 show that an improved separation is obtained by addition of pivalic acid whereas more washings and regulation of the temperature does not have great influence on the separation-Example 29 (S>4-[4'DimethylaminO" 1 -(4' -fluorophenyl)-! -hydroxybutyll-3-hydroxymethyl-benzonitrile To a stirred solution of racemic 4-[4-DimethyIamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-ben2onitrile (0,29 mmol, 100 mg) and vinylbutyrate (3,9 mmol, 0,5 ml) in anhydrous 1,4-dioxane (2,5 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 5 mg). The reaction is heated to 37 °C and followed by HPLC. After 162 hours (at a conversion of 33,9%) the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 72% ee and (S)-diol with 28% ee. Example 30 (S>4-[4-Dimethylamino-1 -(4' -fluorophenyl)-1 -hydroxybuty 1] -3 -hydroxymethy I-benzonitrile To a stirred solution of racemic 4-[4-Dimethylamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzonitrile (0,29 mmol, 100 rag) and vinylbutyrate (0,58 nmiol, 73,6 \|J.l) in-anhydrous 1,4-dioxane (3,0 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 4 mg). The reaction is heated to 37 °C and followed by HPLC. After 194 hours at a conversion of 18,7 %, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are ev^orated iji vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 92% ee and (S)-diol with 14% ee. The reaction was also monitored using 1, 4 and 8 equivalent vinylbutyrate as shown in table 12. Example 31 (S)-4-[4-Dimethylaimno-l-(4'-fluorophenylVl-hydroxybutyl]-3-hydr^ benzonitrile To a stirred solution of racemic 4-[4-Diinethylamino-l-(4-fluoro-pheiiyl)-l-hy(iroxy-butylj-S-hydroxymethyl-benzonitrUe (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 jil) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 5 mg). The reaction is heated to 50 ^^C and followed by HPLC. After 165 hours at a conversion of 30,4 %, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The .combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 98,1% ee and (S)-diol with 30,3% ee. The reaction was also monitored at 25, 37 and 65 °C as shown in table 13. Example 32 (S>4-r4-Dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hy^^ benzonitrile To a stiired solution of racemic 4-[4-Dimethylamino-l-(4-£Iuoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyI-benzonitrile (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 jil, 2, eq.) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 4,5 mg). The reaction is heated to 50 °C and followed by HPLC. After 209 hours, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester and (S)-did- The reaction was also monitored with (200 mg, 500 mg and 1000 mg diol and respectively 150 \|il, 375 [il and 750 \|il vinylbutyrate and 2,85 ml, 2,625 ml and 2,25 ml 1,4-dioxane as shown in table 14. Example 33 (S)-4-[4-Dimethylamino-l-(4'-fluorophenyl)-l"hydroxybutyl]-3-hydroxy^ benzonitrile To a stiired solution of racemic 4-[4-Diniethylamino--l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzonitrile (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 \|xl) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 4 mg). The reaction is heated to 50 °C and followed by HPLC. After 473 hours, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. ^The combined organic phases are ev^orated in vacuo and subsequently analyzed HPLC. The result is shown in table 15. Example 34 (SV4-['4-Dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyn-3-hydroxymethyl-benzonitrile To^ stirred solution of racemic 4-[4-DimethyIamino-H4-fluoro--phenyl)-l-hydroxy" V butyI]-3-hyclroxymethyl-benzomtrile (2,9 mmol, 1000 mg) and vinylbutyrate (5,9 mmol, 750 fxl) in anhydrous 1,4-dioxane (14,25 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 20 mg). The reaction is heated to 50 °C and followed by HPLC. After 139 hours, additional 5 mg lipase was added. After 155 hours, additional 15 mg lipase was added. After 399 hours, at a conversion of 55,7 % a sample was collected. The erLzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. After 560 hours at a conversion of 62,8 %, the reaction was stopped. The enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. The obtained ee-values are shown in table 16. Example 35 (S)-citalopram diol analogues To a stirred solution of racemic citalopram diol analogue (0,29 mmol, 100 mg) and vinylbutyrate (0,29 mmol, 37 \|il) in anhydrous 1-4-dioxane (2,925 ml) is added 4-10 mg PspLL. -The reaction is heated to 50 degrees celcius and followed by HPLC. After -reaction was stopped, the enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 17. Example 36 (S)-4-[4-Dimethylamino-1 -(4' -fluorophenyl)-1 -hydroxybutyl] -3-hydroxymethyl-benzonitrile To a stiired solution of racemic 4-[4-Dimethylamiiio-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-beiizonitrile (29 mmol, 10 g) and vinylbutyrate (58 mmol, 7,5 ml) in anhydrous 1,4-dioxane (142,5 ml) is added lipoprotein lipase pseudomonas sp, (160 U, 250 mg). The reaction is heated to 50 °C and followed by HPLC. After 192 hours at a conversion of 41%, additional 250 mg lipase was added After 504 hours, at a conversion of 63 % the reaction was stopped. The enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Obtained EE-value ((S-diol) = 95% (S-diol/R-diol = 40:1). Example 37 Isolation of (S)-4-[4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyI1-3- hydroxymethyl-benzonitrile by flash-cbromatography A mixture of 372 mg S-citalopram diol with ee-value 94,5% and 628 mg R-citalopram diol butyrate ester with ee-value 59,5% was dissolved in a Tninimtim of ethylacetate/heptane 4:1 with 4% triethylamine. The S-citalopram diol was isolated using flash-chromatography in ethylacetate/heptane 4:1 with 4% triethylamine to obtained 120 mg S-citalopram diol (ee-value 94,5%). Example 38 Isolation of (S)-4-r4-DimethylammO"l-(4'-fluorQphenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile by washing a mixture containing the S-diol base To a mixture of S-citalopram diol and R-citalopram diol butyrate ester (0,3g each) 'was added 24 ml heptane and 66 ml water/methanol (2:1). The clear solution was transferred to a separation fimnel and organic phase was collected. Additional 10 ml heptane was added to aqueous phase and mixed thoroughly. Organic phase was collected. Ekstraction repeated additional 4 times. The combined heptane phases were ekstracted with 20 ml water/methanol (2:1) and aqueous phases were combined and evaporated to half volume and extracted with 3 times 10 ml ethylacetate. The combined ethylacetate phases were dried with Na2S04, filtered and dried in vacuo to obtaine 0,15 g S-citalopram diol containing 2% R-citalopram diol butyrate ester. Example 39 (S)-citalopram diol analogues using Novozymes 435 To a stirred solution of racemic citalopram diol analogue (0,29 mmol, 100 mg) and vinylbutyrate (0,29 mmol, 37 ^1) in anhydrous toluene (2,925 ml) is added 0,2 mg Novozymes 435 and (0,32 mmol, 33 mg) pivalic acid. The reaction is heated to 40 degrees celcius and followed by HPLC. After reaction was stopped, the enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 18. Example 40 The influence of various carboxylic acids on the enzymatic acylation using Novozymes435 To a stirred solution of racemic 4-[4-Dimethylamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzomtrile (0,29 mmol, 100 mg) and vinylbutyrate (0,29 nmiol, 37 ^1) in anhydrous toluene (2,925 ml) is added Novozymes 435, (0,2 mg) and 1,1 eq. Carboxylic acid. The reaction is heated to 40 degrees celcius and followed by HPLC. The enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 19. Example 41 (R>-5-Cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyll hcnzyl butyrate A four-necked flask equipped with stirrer and thermometer was charged with 219 g of a solution containing 21.9 g (0.064 mol) of (±)-4-[4-dimethylamino-l- (4-fluorophenyl)-l-hydroxybutyl]- 3-hydroxymethylben2onitrile in toluene. Then, 14.6 g (0.128 mmol) of vinyl butyrate, 5.07 g (0.128 mol) of pyridine, 7.89 g (0.064 mol) of benzoic acid and 2.19 g of an immobilized enzyme (Novozym 435) were added into the above mixture. The reaction mixture was allowed to warm up to 60 °C and stirred for 15 hours under a slight flow of nitrogen and stirring was stopped. The enzyme was filtrated off from the reaction mixture with Kiriyama funnel, and washed with 50 g of toluene. The combined toluene layei: was washed twice with water (255 ml, 265 ml) and concentrated to give 14.9 g of (R)-5-Cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate benzoic acid salt To a mixture of 4,0 g of (R)-5"Cyano-2- [dimethylamino- (4'-fluoropheiiyl> flydroxybutyl] benzyl butyrate benzoic acid salt (7.48 mmol), 40 ml of water and 40 mi of toluene were added 1.26 g of 30% sodium hydroxide (9.45 mol) with stirring, the mixture was kept stirring for 30 minutes. After stirring, the mixture was allowed to stand and the toluene layer was separated. The toluene layer was washed with 40 ml of water and concentrated at 40 0C under reduced pressure, to thereby obtain 2.7 g of (R)-5-Cyano-2- [dimethylamino- (4-fluorophenyl)-hydroxybutyl] benzyl butyrate. IH NMR (400 MHz. CDC13) a(ppm): 7.69 (IH, s), 7.68-7.56 (2H, m), 7.36-7.23 (2H, m), 7.02-6.90 (2H, m), 5.41 (IH, d, J = 14.9 Hz), 4.99 (IH, d, J = 14.6 Hz), 2.59-2.42 (IH, m), 2.40-2.30 (3H, m),2.26 (2H, dt, J= 7.3 Hz, 1^ Hz), 2.18 (6H, s), 1.68-1.57 (2H, m), 1.67-1.46 (2H, m), 0.93 (3H, t, J = 7.6 Hz). Example 42 Escitalopram,oxalate (iS)-4-(4-Dimethylamino)-1-(4 '-fluorophenyl)-1 -hydroxybutyl)-3-hydroxymethylbenzonitrile (15.8 g, 46.2 mmol) having a chemical purity of 99% and an eeof98.7%asdetenninedby Chiral Supercritical Fluid (Chromatography, was dissolved in toluene (100 mL). Triethylamine (13.0 mL, 93.2 mmol) was added followed by slow addition of a solution of tosyl chloride (9.4 g, 49.4 lomol) in toluene (lOOmL). The resulting solution was stirred at room temperature for 20 min. then added water (50 mL) and cone. Ammonia (25 mL). The mixture was stiired at 40 0C for 2 rnin^ transferred to a separatory funnel. The phases were separated and the organic phase was washed with water (50 mL), dried with MgS04, filtered and concentrated under reduced pressure to give the cmde product (14.2 g) in 95% yield. The crude product was dissolved in ethanol (17 mL) and was added a solution of oxaUc acid (3.95 g, 43.9 mmol) in ethanol (27 mL). The precipitate was collected by filtration to give Escitalopram, oxalate (14.0 g), which was re-crystaUised from ethanol (85 mL) to give the final product (12.2 g). The purity was deteraiined to be 99.65% by HPLC. The ee was 98.5% deteraiined by Chiral Supercritical Fluid Chromatography. WE CLAIM: 1. A method for the preparation of the S- or R-enantiomer of a diol having the formula wherein R is cyano or a group which may he converted to a cyano group, Z is a group -CH2-N(R'R') wherein R'and R' are C1-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may he converted to a dimethylaminomethyl group, the dotted line is a double or single bond and Hal is halogen, or a salt thereof and/or the opposite F enantiomer of an acylated diol having the formula wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R1, wherein R1 is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-10-allcyl)amino, aryl, aryloxy, arylthio and heteroaiyl, or R1 is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Cuio-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, s C1-10-alkyithio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di"(C1-10- aIkyl)amino and Y is a bond, 0, S or NH, or a salt thereof comprising a) subjecting a racemic compound of formula wherein R, Z, the dotted line and Hal are as defined above, to selective enzymatic acylation using an acylating agent having the formula or an isocyanate having the formula R1-N=C=0 or an isothiocyanate having the formula R1-N=C=S; wherein X is O or S; W is O or S; U is O or S, V is halogen; R° is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from CMo-alkoxy, CMQ- • alkylthio, hydroxy, halogen, amino, nitro, cyano^ C1-10-alkylamino, di-(C1-10-alkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R° is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Cuo-allcyl, C1-10-alkenyl, C1-10-allkny1 C1-10-alkoxy, CMQ- alkjdthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(CMo-alkyI)amino; R1 is as defined for R0; R is as defined for R , or R is a suitable leaving group; or R° and R^ together form a chain of 3 to 5 carbon atoms; provided that W and U is not S when X is S; to fonn a mixture of tbe starting material of formula (II) in either the R- or the S-form and opposite enantiomer of a compound having the fonnula wherein R, W, Hal, R3, the dotted line and Z are as defined above; or b) subjecting a racemic compound of formula -1 wherein R, Z, W, Hal, the dotted line and R are as defined above; to selective enzymatic deacylation to form a mixture of deacylated compound of formula wherein R, Hal, the dotted line and Z are as defined above in either the R- or the S-form and the acylated starting material of formula (IV) in the form of the opposite enantiomer; optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (II) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof 2. The method according to claim 1, wherein the acylation of the compound of formula (II) results in a mixture containing the compound of formula (II) in the S-form and the compound of formula (IV) in the R-form. 3. The method according to claim 1, wherein the acylation of the compound of formula (II) results in a mixture containing the compound of formula (II) in the R-form and the compound of formula (IV) in the S-form. 4. The method according to claim 1, wherein the deacylation of the compound of formula (TV) results in a mixture containing the compound of formula (IV) in the S-form and the compound of formula (II) in the R-form. 5. The method according to claim 1, wherein the deacylation of the compound of formula (IV) results in a mixture containing the compound of formula (IV) in the R-form and the compound of fonnula (II) in the S-form. 6. The method according to any of claims 1-5, wherein Hal is fluoro and R is halogen or cyano preferred R is cyano. 7. The method according to any of claims 1-6, wherein the dotted line represents a single bond. 8. The method according to any of claims 1-7 wherein and Z is dimethylaminomethyl or a group that may be converted to a dimethylaminomethyl group, preferred Z is a dimethylaminomethyl group. 9. The method according to any of claims 1, 2, 3 and 6-8, wherein the acylating agent is a compound selected from the compounds of formula (Ilia), (lUb) and (IIIc). 10. The method according to claim 9, wherein the acylating agent is a compound selected from the compounds of formula (Ilia) and (Illb). n. ' The method according to claim 10 wherein the acylating agent is a compound of foraiula(ina). IZ- The method according to claim IQ wherein the acylating agent is a compomid offoraiula(IIIb). 13. Tlie method accordmg to claim ^ wherein the acylating agent is a compomid of fonnula(inc). 14 The method according to any of claims 9 -11 wherein U is O. 15, The method according to any of claims 9-14 wherein W is O. 16. The method according to claims 9-15 wherein X is O. n. The method according to any of claims 9-16 wherein R0, R1 and R2 are independently selected from CM-alkyl, C1-6-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino or R2 is a leaving group, such as succinimidyl, HOBt and ipfp or R^ and R^ together form a chain of 3-5 carbon atoms. \ 2. The method accordii^ to claim 17 wherein R0, R1 and R2 are independently selected from C1-3-alkyl, C2.3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di-(C1-3-alkyl)amino, l9. The method according to claim Ig wherein R0 and R1 are C1-3-alkyl and R2 is C1-3-alkyl substituted one or more times with halogen or R2 is C2-3-alkenyL %p. The method according to claim H wherein R0 and R1 are independently selected from C1-4-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected fiom C1-4-3Jkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(C1-4-alkyl)anxino. Zl, The method according to claim 2D wherein R0 and R1 are independently selected from Ct-s-alkyl, C2.3-alkenyl or C2-3-aIkynyI all of which may all optionally be substituted one or more times with substituents selected from C1-3- alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di- (C1-3'aIkyl)amiiio. ISL. The method according to claim 21 wherein R° and R^ are Ci-i-alkyl. 23. The method of claim 2J,.wherein R^ and R^ are C1-3-aDcyl, smtable methyl, ethyl or propyl, preferred propyl. 24^ ♦ The method of claim 12- wherein R^ and R^ are independently selected from Ci-4-alkyl, C2-4-alkenyl and C2^-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-4-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-allcylamino and di"(C1-6-alkyl)amino or R2 is a leaving group, such as succinimidyU HOBt and pfp. 25: The method of claim 24 wherem R1 is selected from C1-3-alkyl, C2.3-alkenyl or Cao-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cu3-allcylamino and di-(C1-3-alkyl)amino and R2 is Cia-alkyl substituted or more times with halogen or R2 is C2-4-alkenyl. 26 The method according to claim 25"wherein R' is C1-3-alkyl, C1-3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci.3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-aIkylaniino and di-(C1-3-alkyl)amino and R2 is Cu3-allcyl substituted one or more times with halogen or R2 is Cao-alkenyl. 2-T The method according to claim 25'wherein R1 is d-s-alkyl. 23- The method according to claim 27 wherein R^ is C1-3-alkyl, such as methyl, ethyl or propyl and C],3-alkyl substituted one or more times with halogen or R is €2-3-alkenyl, 11. The method according to claim Z8 wherein R^ is vinyl. 3(X The method according to claim 251 wherein R^ is propyl. 31. The method according to claim 13 wherein R^ is C1-6-alkyl, C2--alkenyl and C2^-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-4-aIkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-4-alkylamino and di-(C1-4-aIkyl)amino, HL. The method according to claim 3t wherein R^ is Cua-alkyU Ci-j-alkenyl and C2-3-aJkynyl all of which may optionally be substituted one or more times with C1.3-alkoxy, C1-3-alkylthio. hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino^and di-(C1-3-2ilkyl)amino. 33. The method according to claim 3Zwherein R^ is Ci,3-alkyl, C2.3"alkenyl or C2-3-alkynyl preferred R^ is C1-3-alkyl, such as methyl, ethyl or propyl, prefeired propyl 3ih The method according to any of claims 1, 2-3 and 6-^5 wherein the acylating agent is an isocyanate of foraiula R^-N= 35". The method according to claim 3/^ wherein the acylating agent is an isothiocyanate of the formula R^-N=C=S. 36- The method according to 'claim Jk- wherein the acylating reagent is an isocyanate of formula R^-N==C=0. 37 The method according to any of claims 3/^-36 wherein R^ is C1-4-alkyl, C1-6-alkenyl or Cj^-alkynyl all of wfech may optionally be substituted one or more times with substituents selected from C1-4-alkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(C1-4-alkyl)aniino. 3?- The method according to any of claims 37 wherein R^ is C1-3-alkyl, C2-3-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with C1-3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, C1.3-alkylamino and di-(C1-3-alkyl)aniino. 3^- The method according to claim?»8 wherein R^ is C1-6-alkyl, C2.3-aIkenyl or C2-3-alkynyL 40. The method of claim 3^ wherein R^ is methyl, ethyl, or propyl, preferredpropyl 41. The method according to any of claims 1 j and fy^Z wherein the racemic compound of formula (TV) used is a compound wherein Y is O, or S. 44v The method according to claim 41 wherein the racemic compoimd of formula (TV) is a compound wherein Y is 0. 4S. The method according to claim 41 wherein the racemic compoimd of formula (IV) used is a compound wherein Y is S. 4if- The method according to any of claims 1 ^ and tf- % wherein the racemic compound of foraiula (IV) used is a compound wherein Y is a bond. 45. The method according to any of claims 1 . and i^^t wherein the racemic compound of foraiula (IV) used is a compound wherein Y is NH. 46. The method according to any of claims 4J-4S" wherein R is Ci-10-alkyl, C2-10-alkenyl or C2-10-alkynyI all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-aUcylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino. 47» The method of claim 46 wherein R^ is C1-10-alkyl, C1-10-alkenyl or C2-10-aDcynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano. /f 2. The method according to claim 47 wherein R^ is C1-10-alkyL l^^^ The method according to any of claims l^^nS and g-J^o wherein the enzymatic acylation is performed with a hydrolase, such as a hpase, an esterase, an acylase or a protease, 50. The method according to any of claims l.j^-.^. and 3 ?'4g wherein the enzymatic deacylation is performed with a hydrolase, such as a Upase, an esterase, an acylase or a protease. 51, The method according to any of claims tfff^SO wherein the hydrolase is used in the forai of an immobilized enzyme or Cross-Linked Enzyme Crystal (CLEC) enzymes. 5%, The method according to any of claims Lp^-St wherein the enzyme used is selected from Pseudomonas sp, hpoprotein Upase, Candida antartica Upase B and Thermomyces lanuginosus Upase or a mutant or variant of these enzymes. 53. The method according to any of claims ^9--^wherein the enzyme is Pseudomonas sp. Upoprotein Upase or a mutant or variant of Pseudomonas sp. Upoprotein Upase. 5 If, The method according to claim 5S wherein the enzyme used is Pseudomonas sp. Upoprotein Upase. 557 The niethod according to any of claims/jf^-ica. wherein the enzyme is Candida antartica lipase B or a mutant or variant of Candida antartica lipase B. 56, The method according to claim Si'wherein the enzyme is Candida antartica Upase B. 57. The method according to claim 56 wherein the enzyme is Novo2yme®435. S^, The method according to any of claims i^^-St wherein the enzyme is Theimomyces lanuginosus lipase or a mutant or variant of Theraiomyces lanuginosus lipase. S^^ The method according to claim 5^ wherein the enzyme is Themiomyces lanuginosus hpase. ) 60* The method according to claim SJ wherein the enzyme is Lypozymc^^ TL IM. 6J, The method according to any of claims 1-66 wherein the enzymatic acylation or the enzymatic deacylation is carried out in presence of an organic base or an organic acid or a mixture thereof. 62^ The method according to claim Si A^herein the enzymatic acylation or enzymatic deacylation is carried out in the presence of an organic acid. 63. The method according to claim ^wherein the enzymatic acylation is carried out in presence of an organic acid. 6i^, The method according to any of claims 62^3 wherein the organic acid is an aromatic carboxylic acid or an aUphatic caiboxyUc acid. 6S; The method according to any of claims 61-64 "^^^^^ ^® organic acid is selected from n-propionic acid, iso-propionic acid, n-butyric acid, iso-butyric acid, isO'Valeric acid, 2-ethylbutyric acid, cyclohexanecarboxylic acid, pivalic acid, benzoic acid, p-toluic acid, saUcylic acid and 3-phenylpropionic acid. 6^» The method according to claim 6& wherein the organic acid is pivalic acid. 67 A method for the isolation and purification of the compound having the formula wherein R is cyano or a group which may be converted to a cyano group, the dotted hne represents a double or single bond, Hal is halogen, Z is a group -CH2-N(R'R') wherein R'and R' are C1-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a groiq) which may be converted to a dimethylaminomethyl group, W is O or S and R"^ is -Y-R^ wherein Y is a bond, O, S or NH and R^ is C1-10-alkyl C1-10-alkenyl or C2-1Q-alkynyl all of which may optionally be substituted with one or more substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci. lo-alkylamino, di and/or the diol of formula wherein R, Z, Hal and the dotted line are as defined above, or a salt thereof^ from a mixture containing the compound of formula (IV) and the diol of formula (II), which comprises: a) treating said mixture contaimng the compound of formula (IV) and the diol of formula (II) in a mixture of water and an organic solvent in the presence of an acid; b) separating the aqueous phase containing the diol of fonnula (H) as a salt of said acid from the organic phase to obtain an organic phase containing the compound of fonnula (IV) as a salt of said acid; and optionally isolating the compound of formula (II) as the base or as a salt thereof and optionally isolation of the compound of formula (IV) as the base or a salt thereof. 6^, The method according to claim f>1 wherein the S-enantiomer of the diol of formula (H) is separated from the R-enantiomer of the acyl derivative of formula (IV). Cn- The method according to claim gy wherein the S-enantiomer of the acyl derivative of fonnula (IV) is separated from the R-enantiomer of the diol of formula (n). 10« The method according to claims ft7-0 wherein the mixture of the R- or S-enantiomer of an acyl derivative of fonnula (IV) and the opposite enantiomer of an diol of fonnula (II) used in step a) has been prepared according to the method in claims 1-62 71. A method for isolation and purification of flie acyl derivative having the fonnula wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line represents a double or a single bond, Z is a group -CH2-N(RTI'0 wherein R'and R" are Ci-6-aIkyl, or R'and R" araconnected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl groitp, W is O or S; and R^ is -Y-R^ wherin Y is a bond, 0, S or NH and R^ is C1-4o-alkyl, C2-10-aUcenyl or C2-lo-alkynyl all of which may optionally be substituted one or more times with substituents selected from Cj.io-alkoxy, Ci-10-allsyltiuo, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-10-aIkyI)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-4O-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-4o-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cj.jo-aUcylamino and di-(C1-4o-alkyl)amino, or a salt thereof and/or the diol of formula wherein R, Hal, Z and the dotted line are as defined above, from a mixture containing the acyl derivative of fonnula (TV) and the diol of formula (II), which comprises: a) treating said mixture containing the acyl derivative of formula (TV) and the diol of fonnula (U) in a mixture of water, a protic organic solvent and an apolar organic solvent; b) separating the aqueous phase containing the diol of formula (II), from the organic phase to obtain an organic phase containing the acyl derivative of formula (TV); and optionally isolating the diol of fonnula (IT) and/or the compound of formula (IV) from the aqueous/organic ohase, and optionally conversion of the compound of foraiula (H) and /or (TV) to salt thereof. 72. The method according to claim 71 wherein the S-diol of the diol of fonnula (II) is separated from the R-enantiomer of the acyl derivative of formula (IV). 73. The method according to claim 7j wherein the S-enantiomer of the acyl derivative of fonnula (IV) is separated from the R-enantiomer of the diol of fonnula (H). 74. The method according to claims IhTS wherein the mixtuxe of the R- or S- enantiomer of a compound of formula (IV) and the opposite enantiomer of a compound of formula (I[) used in step a) has been prepared according to the method in claims 1-62 7S- A process for the preparation of escitalopram having the formula or a phannaceutically acceptable salt thereof comprising preparation of the S-enantiomer of a diol having the formula wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond, Z is a dimethylaminomethyl group or a group which may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof, or the S-enantiomer of an acylated diol having the formula wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R\ wherein R^ is C1-10-alkyl, C2-10-alkenyl or C2-10-alkyiiyl all of which may optionally be substituted one or more times with substituents selected firom C1-4O-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(Ci-i(ralkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected firom C1-10-alkyl, C1-10-alkenyl, C2-i(ralkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, O, S or NH, or a salt thereof, by the method in any of claims 1-62, optionally followed by, in either order, conversion of the group R to a cyano grqup, conversion of the group Z to a dimethylaminomethyl group, reduction of a double boiid represented by the dotted line to a single bond and /or conversion of the group Hal to a fluoro group and then ringclosure under basic conditions of the S-enantiomer of foraiula (lis) or (TVs) or a labile ester derivative thereof to form a compound of foraiula ^ followed by in either order, if R is not cyano conversion of the group R to a cyano groiq), if Z is not dimethylaniinomethyl conversion of the group Z to a dimethylaminomethyl group, if the dotted line represents a double bond reduction to a single bond and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolation of escitalopram or a pharmaceutically acceptable salt thereof. 7^ The method according to claim 7S wherein the rrdxture of the R- or S-enantiomer of a compound of formula (H) and the opposite enantiomcr of the compound of formula (IV) obtained by emymatic acylation has been separated from each other by the isolation and purification process according to any of claims 84-87. 77 - The method according to claim is" wherein the R- or S-enantiomer of a compound cf formula (H) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 88-91, -^^ A method for the preparation of racemic citalopram and /or escitalopram or a pharmaceutically acceptable salt thereof comprising preparation of the R-enantiomer of a diol having the formula wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond and Z is a dimethylaminomethyl group or a group wl-vich may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof. or the R-aiantiomer of an acylated diol having the formula wherein R, Z, the dotted line Hal are as defined above, W is O or S, and R^ is -Y-R\ wherein R^ is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom C1-10-alkoxy, C1-4O-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-4o-alkyl)amino, aryl, aryloxy, arylthid and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt thereof, by the method according to any of claims 1-62, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and then ringclosure under acidic conditions of the R-enantiomer of formula (Ilr) or (TVs) to form a mixture of the S-enantiomer of the compound of formula and a minor amount of the corresponding R-enantiomer, followed by, in either order, if R is not cyano conversion of the group R to a cyano group and, if Z is not dimethylaminomethyl conversion of the group Z to a dimethylaminomethyl group, if the dotted line represents a double bond reduction to form a single bond and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolatiC1-4 of escitalopram and/ or racemic citalopram or a phaimaceuticaliy acceptable salt thereof. 79. The method according to claim 78 wherein the racemic citalopram is isolated by precipitation of racemic citalopram free base or a salt thereof and escitalopram is recovered from the mother liquor of the precipitation. 80* The method according to claim "7^ wherein the mixture of the R- or S-enantiomer of a compound of formula (D) and the opposite enantiomer of the compoimd of formula (IV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 84-87. 8) The method according to claim 7^ wherein the R- or S-enantiomcr of a compound of formula (II) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 88-91.

Full Text

The present invention relates to a novel method for the preparation of optically active intermediates useful for the preparation of escitalopram involving selective enzymatic acylation or deacylation.
Background of the inventioa
Citalopram is a well-known antidepressant drug that has now been on the market for some years.
It is a selective, centrally acting serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitor, accordingly having antidepressant activities.
Citalopram was first disclosed in DE 2,657,013, corresponding to US 4,136,193. This patent pubUcation i.a. outlines a process for preparation of citalopram jfrom the corresponding 5-bromo-derivative by reaction with cuprous cyanide in a suitable solvent and by alkylation of 5-bromo-phtalane.
US Patent No 4,943,590 corresponding to EP-B1-347 066 describes two processes for the preparation of escitalopram (S-enantiomer of citalopram).
Both processes use the racemic diol having the fomiula

as starting material. According to the first process, the diol of formula (I) is reacted with an enantiomerically pure acid derivative, such as (+) or (-)-a-methoxy-a-trifluoromethyl-phenylacetyl chloride to forai a mixture of diastereomeric esters, which are separated by HPLC or firactional crystallization, whereupon the ester with the correct stereochemistry is enantioselectively converted into escitalopram. According to the second process, the diol of formula (11) is separated into the enantiomers by stereoselective crystalhzation with an enantiomerically pure acid such as (+)-di-p-toluoyltartaric acid, whereupon the S-enantiomer of the diol of the formula (A) is enantioselectively converted to escitalopram.
Escitalopram has now been developed as an antidepressant. Hence, there is a desire for an improved method for preparation of escitalopram.
It has now been found that the S-enantiomer of the diol of formula (I) above as well as acylated derivatives thereof may be prepared by selective enzymatic acylation of the primary hydroxyl group in the racemic diol to obtain S-diol or an acylated derivative thereof with high optical purity and fiirther that the enantiomers obtained may be separated by a series of isolation and purification operations.
The invention
Accordingly, the present invention relates to a novel process for the preparation of the S- or R-enantiomer of a diol having the formula

wherein R is cyano or a group which may be converted to a cyano group, Z is a group -CH2-N(R'R") wherein R'and R" are C1-6-alkyl, or R'and R" are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl group, the dotted Une represents a double or a single bond and Hal is halogen or a salt thereof, and/or the opposite enantiomer of an acylated did having the formula

wherein R," Z, the dotted Une and Hal are as defined above, W is O or S, and R^ is -Y-R', wherein R^ is C1-10-alkyl, C2-10-aIi:enyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from CMO-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(CMo-allcyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl; wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt thereof, comprising
a) subjecting a racemic compound of formula

or R and R together form a chain of 3 to 5 carbon atoms;
provided that W and U are not S when X is S; to form a mixture of the starting material of foraiula (II) in either the R- or the S-form and the opposite enantiomer of the acylated diol having the formula
wherein R, W, Hal, R"*, the dotted hne and Z are as defined above; or b) subj ecting a racemic compound of formula

wherein R, Z, W, Hal, the dotted line and R3are as defined above; to selective enzymatic deacylation to form a mixture of deacylated compound of formula

wherein R, Hal, the dotted line and Z are as defined above in either the R- or the S-fonn and the acylated starting material of formula (TV) in the form of the opposite enantiomer;
optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (H) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof.
The invention also relates to methods for the separation of mixtures of an enantiomer of fonnula (IV) from the opposite enantiomer of formula (11) and to the R- and S-enantiomers of the compounds of formula (TV) above.
Finally, the invention relates to a method for the preparation of escitalopram and racemic citalopram from the enantiomers of a compound of formula (II) obtained by the selective enzymatic acylation or deacylation according to the invention, or the enantiomers of the optically active acyl derivative of formula (IV) obtained by the selective enzymatic acylation or deacylation according to the invention.
Detailed description of the invention
Wlien used in connection with the compounds of formula (11), (TV) and (V), the terms "enantiomer", "R-enantiomer", "S-enantiomer", "R-form", "S-form", "R-diol" and * ' "S-diol" refer to the orientation og the groups around the carbon atom to which the 4-Hal-phenyl group is attached.

ine present invention thus relates in one embodiment to a selective enzymatic acylation as above and in another embodiment to selective enzymatic deacylation as above.
Selecrive enzymatic acylation means that the'enzymatic acylation is preferentially effective for conversion of one of the enantiomers of a compound of formula (II) preferentially leaving the other enantiomer of the compound of formula (II) unconverted in the reaction mixture.
Selective enzymatic deacylation means that the enzymatic deacylation is preferentially effective for conversion of one of the enantiomers of a compoimd of formula (IV), preferentially leaving the other enantiomer of the compound of formula (IV) unconverted in the reaction mixture.
The selective acylation according to the invention thus results in a mixture containing preferentially the compound of formula (II) in the S-form and the compound of formula (IV) in the R-form, or it may result in a mixture containing preferentially the compound of formula (E) in the R-form and the compound'of formula (IV) in the S-form.
Likewise, the selective enzymatic deacylation may result in a mixture containing preferentially the compoimd of formula (IV) in the S-fonn and the compound of formula (E) in the R-forai, or it may result in a mixture containing preferentially the compound of formula (IV) in the R-form and the compound of formula (H) in the S-form.
The composition mixture obtained after acylation or deacylation according to the invention depend on the specific hydrolase used and the conditions under which the reaction is carried out. Characteristic of the enzymatic acylation/deacylation according to the invention is that a considerably larger portion of one enantiomer is converted than of the other. The optical purity of the diol of formula (11) and/or the acylated compound of formula (IV) obtained by the optical resolution method of the present invention is usually at least 90% ee, preferably at least 95% ee, more

preferably at least 97% ee and most preferably at least 98% ee. However, lower values for the optical purity are acceptable.
The starting material for the enzymatic method of the invention is a racemic diol of formula (II) or a racemic acyl derivative of formula (TV)
In a preferred embodiment of the invention R is halogen or cyano, most preferred cyano-
In a further preferred embodiment of the invention Hal is fluoro,
In a further embodiment of the invention, the dotted line in formula (II) and (IV) is a single bond.
In one embodiment Z is dimethylaminomethyl or a group that may be converted to dimethylaminomethyl. In a suitable embodiment Z is dimethylaminomethyl.
Most preferred, Hal is fluoro, R is cyano, the dotted line is a single bond and Z is dimethylaminomethyl.
The acylatmg agent used for the encymatic acylation according to the invention may suitable be one of the compounds of formula (Dla), (lUb) and (Hie).
In another embodiment the acylating agent used according to the invention is any of the compounds of formula (Ula) and (Illb).
According to a further embodiment of the invention, the acylating agent used is a compoimd of formula (IHa).
According to a still another embodiment of the invention, the acylating agent used is a compound of formula (Hlb).
According to another embodiment of the invention the acylating agent is a compound offonnula(inc).

When tlie acylating agent is a compound of foraiula (Ilia), U is suitable O. When the acyalting agent is any of the above, W is suitable 0.
When the acylating agent is any of the above, X is suitable O.
Suitable, the substituents R°, R1and R2in any of the acylating agents (Ilia), (UJb) and (Hie) defined in any of the embodiments above are as follows: R°, R1and R2are independently selected from C1-6-alkyi, C2-6-alkenyl and C1-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(CiHS-aIkyl)amino or R2is a suitable leaving group, such as succinimidyl, HOBt and pfp, or R0 and R1 together form a chain of 3-5 carbon atoms more suitable R0, R1 and R2 are independently selected from C1-4-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, uitro, cyano, C1-6-alkylamino and di-(CM-alkyI)amino or R^ is a suitable leaving group, such as succinimidyl, HOBt and pfp or R° and R^ together forai a chain of 3-5 carbon atoms, more preferred R0, R1 and R2 are independently selected from C1-3-alkyl, C2.3-aUcenyl and C2-3-al1 Suitably, the substituents R° and R1 in acylating agents of formula (IHa) as defined in any of the embodiments above, are as follows:
R° and R1 are independently selected from C1-6s-alkyl, C2.6-allcenyl and C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from C].6-aUcoxy, C1-6 alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-allcylamino and di-(C1-6-alkyl)amino, more suitable R0 and R1 are independently selected from CM-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, CM-

alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-{CM-alkyl)amino, preferably R^ and R* are independently selected from Cio-alkyl, C2.3-alkenyl and Cao-alkynyl all of which may all optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyauo, Ci.3-alkylamino and di-(Ci.3-alkyl)aniino, more preferred R° and R^ are independently CM-alkyl and most preferred, and most preferred R^ and R^ is Ci.3-alkyl, in particular unbranched Cio-^alkyl, suitable propyl.
Suitably, the substituents R^ and R^ in acylating reagents of formula (lUb) as defined in any of the embodiments above, are as follows:
R^andR^ are independently selected from C1-6-alkyl, C2-6-alkenyl and Ci-a-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci-6-alkyl)amino or R is another leaving group, such as succinimidyl, HOBt and pfp, more suitable R and R are independently selected from C1-6-alkyl, Ca^-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-allcylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino or R^ is another leaving group, such as succinimidyl, HOBt and pfp, preferably R^ is selected from Ci.3-alkyl, C2.3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1.3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cio-alkylamino and di-(C1-3-alkyl)amino and R^ is CM-alkyl substituted one or more times with halogen, R" is C2^-alkenyl or R^ is another leaving group, such as succinimidyl, HOBt and pfp, more preferred R^ is Ci.3-alkyl, C2.3-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1.3-allcoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di-(Ci.3-all
~\ 1
R^ meaning C1-6-alkyl substituted one or more times with halogen is a suitable leaving group including groups such as 2,2,2-trichloroethyl and 2,2,2-trifluoroethyl, in particular 2,2,2-trifIuoroethyL
In a specific embodiment of the invention, the acylating agent of formula (mb) as above is a compound wherein R is vinyl.
According to another specific embodiment of the invention, the acylating agent of foraiula (mb) is a compound as above wherein R^ is propyl. This specific embodiment covers a preferred acylating agent of the invention, namely vinyl butyrate.
According to a fiuther embodiment of the invention, the acylating agent is a compound of formula (Hie).
Suitably, the substituent R^ in the compound of formula (IIIc) as defined in any of the embodiments above, are as follows:
R' is Ci.6-alkyl, C2-6-alkenyl and C2^-alkynyI all of which may optionally be substituted one or more times with substituents selected from C1-6-alkoxy, Ci- V is suitable chloro.

The acylating agent used according to the invention may also be an isocyanate of formula R^-N=C=0 or an isothiocyanate of the formula R^-N=C=S.
Thus, in a further embodiment of the invention, the acylating agent is an isothiocyanate of the formula R^-N=C=0,
According to another embodiment of the invention, the acylating agent is an isocyanate of formula R^-N=C=S.
Suitably, the substituent R^ in the isocyanate and the isothiocyanate as defined in any of the embodiments above, are as follows:
R^ is Ci-6-alkyl, Ca-e-alkenyl or C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected firom Ci-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-alkyl)amino, more suitable R' is Cu-alkyl, Cz^-allcenyl or C1-6i-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-4-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(CM-alkyl)amino, preferebly R* is Ci.3-alkyl, Czo-alkenyl or Cao-allcynyl all of which may optionally be substituted one or more times with C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-aikylamino and di-(C 1-3-alkyl)amino, more preferred R* is C1-3-alkyl, C2-3-alkenyl or C2-3-alkynyl, and more suitable R^ is C1-3-alkyl, in particular unbranched C1-6-alkyl, such as methyl, ethyl or propyl
The invention also covers a method for selective en2ymatic deacylation of a racemic compound of formula (IV) as defined above.
Suitable, tlae racemic compound of formula (TV) used, is a compound wherein Y is O, orS.
According to a further embodiment the racemic compound of formula (TV) used, is a compound wherein Y is O.
In still another embodiment of the invention the racemic compound of formula (IV) used is a compound wherein Y is S.

In another embodiment of the invention the racemic compound of formula (IV) used is a compound wherein Y is a bond.
Suitably, the substituent R' in the racemic compound (IV) as defined m any of the embodiments above, is as foUows: Cio-alkyl, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected fi-om C1-10-alkoxy, Ci.io-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMO-alkylamino and di-(C1-10-aIkyl)amino, more suitable R' is Ci.io-alkyl, C1-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano, preferably R' is C1-10-alkyl, preferably unbranched Ci.io-alkyl and more preferred R' is unbranched C4-io-aIkyl.
According to the invention, selective enzymatic acylation is carried out under conditions substantially suppressing hydrolysis. Hydrolysis, which is the reverse reaction of the acylation reaction, takes place if water is present in the reaction system.
Thus, selective enzymatic acylation is preferably carried out in a water-free organic solvent or ahnost anhydrous organic solvent ( enzymes normally require the presence of some water to be active). The examples below, illusfrate how addition of water affects conversion. The percentage of water allowed in a particular reaction system, may be determined by a person skilled in the art.
The organic solvent which may be used for the acylation reaction, is not particularly important as long as it does not deactivate the enzyme used. Suitable solvents include hydrocarbons such as hexane, heptane, benzene and toluene; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, 1,4-dioxane, tert-butyl mediyl ether and dimethoxyetlaane; ketones such as acetone, diethyl ketone, butanon, and methyl ethyl ketone; esters such as methyl acetate, ethyl acetate, ethyl butyrate, vinyl butyrate and ethyl benzoate; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane; seconday and tertiary alcohols, such as tert-butanol; nitrogen-containing solvents such as dimethylformamide, acetoamide, formamide.

acetonitrile and propionitrile; and orotic polar solvents such as dimethylsulfoxide, N-methylpyrrolidone and hexamethylphosphorous triamide.
Among them, hydrocarbons such as hexane, heptane, benzene and toluene, ethers such as diethyl ether, diisopropyl ether, 1,4-dioxane and tert-butyl methyl ether and esters such as vinyl butyrate, are preferred. For one enzymes the most preferred solvents may be aromatic hydrocarbons such as benzene or toluene and ethers, most preferred toluene and for another enzyme the most preferred solvents may be ethers such as 1,4-dioxane (see the examples below). The above solvents may be used singly or in a combination of two or more solvents.
The concentration of racemic diol of formula (II) and acylating agent should not be too high as a high concentration of reagents in the solvent may lead to non-selective acylation of the racemic diol. Suitable the concentration of racemic diol and acylating reagent is each below 1,0 M, more suitable below 0,5 M, even more suitable below 0,2 M or even more suitable below 0,1 M. A person skilled in the art will be able to determine the optimal concentration of racemic diol and acylating agent.
Selective enzymatic deacylation is preferably carried out in water or a mixture of water and an organic solvent, suitable in presence of a buffer. The organic solvent which may be used in the reaction, is not particularly important as long as it does not deactivate the enzyme used. Suitable organic solvents are solvents miscible with water such as alcohols, acetonitrile, DMF, DMSO, dioxane, DME and diglyme. The skilled person will be able to identify other suitable solvents. A person sldlled in the art will be able to determine the optimal concentration of racemic compound of formula (IV) used in the reaction
The stereoselectivity of the enzyme used, may be increased by carrying out the acylation or deacylation in presence of an organic acid and /or an organic base.
Accordingly, the present invention also relates to a process wherein the enzymatic acylation or the enzymatic deacylation is carried out in presence of an organic base or an organic acid or a mixture thereof

In a particular embodiment, the invention relates to a process wherein the enzymatic acylation or enzymatic deacylation is carried out in the presence of an organic acid, suitable an organic carboxyUc acid.
In a further embodiment, the enzymatic acylation is carried out in presence of an organic acid, suitable an organic carboxyUc acid.
Suitable the above mentioned organic acid is an aromatic carboxyUc acid or an aliphatic carboxyUc acid.
As an organic acid which may be used in the reaction, there may be mentioned, alkyl carboxyUc acids, cycIoalkylcarboxyUc acids, cycloalkylalkylcarboxyUc acids, optionally substituted phenyl-alkylcarboxylic acids and optionaUy substituted phenylcarboxyhc acids. Suitable aUphatic carboxyUc acids, are carboxyUc acids such as formic acid, acetic acid, propionic acid, n-butyric acid, iso-butyric acid, 2-ethylbutyric acid, n-valeric acid, iso-valeric acid, pivaUc acid, n-caproic acid, iso-caproic acid, decanoic acid, crotonic acid, paknitic acid, cyclopentanecarboxyUc acid, cyclohexanecarboxyUc acid, phenyl-CM-alkylcaiboxyUc acids such as 3-phenylpropionic acid, 4-phenyIbutyric acid, oxaUc acid, malonic acid and tartaric acid. Suitable aromatic carboxyUc acids, includes acids such as benzoic acid, p-chlorobenzoic acid, p-nitrobenzoic acid, p-methoxybenzoic acid, p-toluic acid, o-toluic acid, m-toluic acid, naphthoic acid, phthaUc acid and terephthaUc acid, saUcyUc acid, hydrocinnamic acid for instance.
Thus, according to one embodiment of the invention, the organic acid used to improve stereoseleselectivity of the enzyme is selected from n-propionic acid, iso-propionic acid, n-butyric acid, iso-butyric acid, iso-valeric acid, 2-ethylbutyric acid, crotonic acid, palmitic acid, cyclohexanecarboxyUc acid, pivaUc acid, benzoic acid and;?-toluic acid, saUcylic acid and 3-phenylpropionic acid. According to a further embodiment of the invention, the carboxyUc acid used is pivaUt acid.
The amount of the organic acid to be used is not particularly restricted, but the molar ratio relative to a substrate is usually 0.1 to 10, preferably 1.0 to 3.0, and more preferably 1.0 to 2.0.

Altematively, a tertiary amine may be used to improve selectivity of the enzyme, either alone or together with any of the above mentioned organic acid. As suitable organic base there may be mentioned, triethyl amine, pyridine, 4-dimethylaminopyridine and pyridine is preferred. Suitable combinations of organic acid and organic base are benzoic acid and pyridine for example.
The amount of the tertiary amine to be used is not particularly restricted, but the molar ratio relative to a substrate is usually 0.5 to 3.0, and preferably 0.5 to 2.0.
The enzymatic acylation or deacylation according to the invention is carried out using a hydrolase, such as a lipase, an esterase, an acylase or a protease.
Thus, according to one embodiment of the invention, enzymatic acylation is performed with a hydrolase, such as a lipase, an esterase, an acylase or a protease. The enzymes useful according to the invention are such enzymes capable of performing R-selective acylation or S-selective acylation of the primary hydroxy group in the racemic compound of formula (II).
According to another embodiment of the invention, enzymatic deacylation is performed with a hydrolase, such as a lipase, an esterase, an acylase or a protease. The enzymes useful according to the invention are such enzymes capable of performing R-selective deacylation or S-selective deacylation of the acyl group in the racemic compound of formula (IV).
As used herein "hydrolase" and "enzyme" either generally or in relation to a specific enzyme, means not only the enzyme itself, but also cultured products containing the enzyme, such as culture fluid containing a cell body, or a cultured cell body, and processed product of the cultured product (for example a crude extract, a freeze-dried micrcorganism or cell, an acetone dried microorganism or cell, a ground product of such microorganism or cell, or the like).
Additionally, the "enzyme" or "hydrolase" may be immobilized as the enzyme itself or as a cell body by loiown techniques, and may be used in immobilized form. The

immoDiiizauon may oe camea out by metnods Known to me person siaiied m the art, such methods include, for example carrier bonding, cross linking, encapsulation and the like-Thus, in one embodiment of the invention the hydrolase is used in the form of an immobilized enzyme or Cross-Linked Enzyme Crystal (CLEC) enzymes.
It has been found that enzymatic acylation accordmg to the invention may be carried out using Novozyme 435, from Candida antartica, LipoZyme TLIM from Thermomyces lanuginosus or Lipoprotein Lipase pseudomonas sp. (isolated from Pseudomonas Cepacia and obtained from Fluka), and particularly good results have been found when using Novozyme 435, from Candida antartica or Lipoprotein Lipase pseudomonas sp.
Thus, according to one embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein Upase, Candida antartica Upase B or Thermomyces lanuginosus lipase.
According to another embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein lipase or Candida antartica lipase B.
As mentioned above, use of one of the above mentioned enzymes according to the invention also covers the use of cultured products containing the enzyme, such as culture fluid containing a cell body, or a cultured cell body, processed product of the cultured product and any immobilized forms of these enzymes/cultured products.
The use of any of the above specifically mentioned enzymes according to the invention also covers the use of mutants, variants or any equivalents of the above specifically mentioned enzymes, which are capable of performing the selective acylation or deacylation according to the invention. The variants or equivalents thereof may be isolated from various strains of Pseudomonas, Candida or Thermomyces, or any other source, or they may be prepared by mutation of the DNA encoding the above mentioned enzymes leading to variations in the amino acid composition of the enzyme. Suitable the mutants or variants of the above mentioned

enzymes axe variants and mutants where single amino acids have been removed or replaced by other amino acids, and suitable the amino acid sequence of the variant or mutant is more than 60 % identical, preferably more than 80 % or most preferred more than 90 % identical to the above mentioned enzymes.
Thus, according to one embodiment of the invention, the enzyme used is Pseudomonas sp. lipoprotein Upase or a mutant or variant thereof. Preferably Pseudomonas sp. lipoprotein Upase is used.
According to another embodiment of the invention, the enzyme used is Candida antartica lipase B or a mutant or variant thereof. Preferably, the enzyme used is Candida antartica Upase B.
According to a further embodiment of the invention the enzyme is Novozyme®435 (Candida antartica Upase B immobiUzed on acrylic resin, available from the company Novozymes A/S).
According to still another embodiment of the invention, the enzyme used is Thermomyces lanuginosus Upase or a mutant or variant thereof. Preferably the enzyme used is Thermomyces lanuginosus Upase.
According to a further embodiment of the invention, the enzyme used is Lipozyme^^ TLIM, also available from the company Novozymes A/S.
The preferred reaction conditions for enzymatic acylation/deacylation differ depending on the particular enzyme used, whether it is immobiUsed or not etc.
A suitable temperature for the reaction lies between 0-80 °C, more preferably between 20-60 °C, or more preferred between 30-50 °C.
The amount of enzyme to be used is not particularly restricted, but is usually O.Ol-I.O,
nrftferahlv 0.02-0.5 and mnrf* nr^^f^araKU/ 0 09-0 1 ^c \x/f*iaVit ratin r^lativp tn CTiKctroto

The reaction may be carried as a batch process or it may be carried out as a continous process. The enzyme may be used in a plurality of batches repeatedly or continuously. The reaction time is not particularly restricted, and will depend on the enzyme used and the scale and type production method (batch or continues).
The present invention also relates to an S- or R-enantiomer of a compound having the foraiula (TV)

wherein R, Hal, R"', W, the dotted line and Z are as defined above,or a salt thereof
According to one embodiment of the invention, the optically active acyl derivative above is the S-enantiomer. According to another embodiment of the invention, the optically active acyl derivative above is the R-enantiomer.
According to one embodiment of the invention the R- or S-enantiomer above is a compound wherein R is halogen or cyano, preferably R is cyano.
According to a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Hal is fluoro.
According to a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein the dotted line represents a single bond.

In another embodiment of the invention, the R- or S-enantiomer above is a compound wherein Z is dimethylaminomethyl or a group that may be converted to dimethylaminomethyi, more suitable Z is dimethylaminomethyl, and the other substituents are as defined above.
In another embodiment of the invention, the R- or S-enantiomer above is a compound wherein Z is dimethylaminomethyl, Hal is fluoro, the dotted line represents a single bond, and R is cyano or halogen, suitable cyano.
In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is 0, or S, preferably Y is O and the other substituents are as defined above. In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is S and the other substituents are as defined above.
In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is a bond and the otlier substituents are as defined above.
In a further embodiment of the invention, the R- or S-enantiomer above is a compound wherein Y is NH and the odier substituents are as defined above.
Suitable, the R- or S-enantiomer as defined in any of the embodiments above is a compound wherein R^ is C1-6-alkyl, C2-6-alkenyl or C2-6-alIcynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(C1-6-alkyl)amino, more suitable R' is CM-alkyl, C1-6-allcenyl or C1-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(CM-alkyl)amino, preferably R^ is C1-3-alkyl, Ca-s-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with substituents sefected from Ci.3-alkoxy,'C1-3-allcylthio, hydroxy, halogen, amino, nitro, cyano, Cio-alkylamino anddi-(Ci,3-alkyl)aniino and most preferred R* is Cio-allcyi, in particular imbranched Ci,3-alkyL

According to another suitable embodiment of the invention, the R- or S-enantiomer as defined in any of the embodiments above is a compound wherein R^ is as follows: Ci. lo-alkyi, C2-10-aIkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times ^^'ith substituents selected from Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci- After the completion of the acylation reaction or the deacylation reaction, an enantiomer of the diol derivative represented by the formula (II) is obtained as a -mixture with the opposite enantiomer of the compound of formula (IV), This reaction mixture is then optionally separated from the enzyme.
In order to obtain the desired enantiomer of formula (11) and/or (IV) with high chemical purity, it is necessary to separate it form the opposite enantiomer of formula (TV) and (II), respectively, in an efQcient manner. However, it is difficult to efficiently separate the enantiomers by generally known methods because the structure of the acyl derivative of formula (TV) is so similar to the stmcture of the diol of formula (n).
The present inventors have made intensive investigations and have as a result found that the enantiomer of a compound of formula (H) in the form of a salt with an acid is efficiently distributed into an aqueous layer and the opposite enantiomer of a compound of formula (IV) in the form of a salt with the acid is efficiently distributed into an organic layer when the reaction mixture is treated with a mixed solvent containing organic solvent and water in tlie presence of an acid.
Thus, according to another embodiment, the invention relates to a method for the isolation and purification of an acyl derivative having the formula

wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line is a double or a single bond and Z is a group -CH2-N(R'R') wherein R'and-R-' C1-6-alkyl, or R'and R" are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethyiaminomethyl group, W is O or S and R^ is -Y-R^ wherein Y is a bond, 0, S or NH and R^ is C1-10-allcyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted with one or more substituents selected from Ci-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci. lo-alkylamino, di-(C1-10-allcyl)ainino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Ci-10-alkyl, C1-10-alkenyl, CJAQ-alkynyl, Ci-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMO-alkylamino and di-(Ci.io-alkyl)amino, or a salt thereof and/or a diol of formula

wherein R, Hal, the dotted line and Z are as defined above, or a salt thereof, from a mixture containing a compound of fonnula (IV) and a diol of formula (II), which comprises:
a) treating said mixture containing the acyl of formula
(IV) and the diol of formula (II) in a mixture of water and an organic solvent in the presence of an acid;
b) separating the aqueous phase contaming the diol of formula (E) as a salt of
said acid from the organic phase to obtain an organic phase containing the acyl
derivative of fonnula (TV) as a salt of said acid;
optionally isolating the compound of formula (II) as the base or as a salt thereof and optionally the compound of fonnula (IV) as the base or as a salt thereof
According to one embodiment of the invention R is halogen or cyano, preferably cyano in the method for isolation and purification above.
According to another embodiment of the invention Hal is fluoro in the method for isolation and purification above.
According to a preferred embodiment, the dotted line represents a single bond.
In another embodiment of the invention, Z is dimethylaminomethyl or a group that may be converted to a dimethylaminomethyl group. Suitable Z is dimethylaminomethyl.
According to a preferred embodiment of the invention Hal is fluoro, Z is dimethylaminomethyl, the dotted line is a single bond and R is cyano or halogen, preferably cyano.
In a further embodiment of the invention the compound of formula (TV) in the method for isolation and purification above, is a compound wherein Y is O, or S, preferably Y is O and the other substituents are as defined above. In a fimher embodiment of the invention tlae compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is S.

In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is a bond and the other substituents are as defined above.
In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound wherein Y is NH and the other substituents are as defined above.
In a preferred embodiment of the invention, the compound of formula (TV) in the method for isolation and purification above, is a compound wherein R^ is Ci-6-alkyl, C2-6-alkenyl or C2-6-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, Ci.6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-allcyl)amino, more preferred R^ is C1-4-alkyl, C2-4-alkenyl or C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino, preferably R^ is Cj.3-allcyl, Ci-s-alkenyl and Cio-aUcynyl all of which may optionally be substituted one or more times with substituents selected from Ci.3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-a-alkylamino and di-(C1-3"alkyl)amino and most preferred R is Ci.3-alkyl, in particular unbranched C1-3-alkyl.
In another preferred embodiment of the invention, the compound of formula (TV) in the method for isolation and purification above, is a compoimd wherein R^ is as follows: C1-10-alkyl, C2-10-alkenyl or C1-10-allcynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, CMQ-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-6-allc/lamino and di-(CMo-alkyl)amino, more suitable R* is C1-10-alkyl, C1-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano, preferably R* is C1-10-aUcyl, preferably unbranched C1-10-alkyl and more preferred R^ is unbranched C4.io-alkyl.
By the above method for isolation and purification, a salt of the R- or S-enantiomer of a compound having4he formula (II) with an acid may be selectively extracted into an

aqueous layer and a salt of the opposite enantiomer of a compound of formula (TV) with an acid may be selectively separated into an organic layer by treating, in the presence of the acid, a mixture comprising an enantiomer of formula (II) and the opposite enantiomer of formula (TV), reaction solvent and the like with an organic solvent and water, and in presence of an acid. In this case, the salt of the enantiomer of formula (IV) and the salt of the enantiomer of formula (II) may be separated with little loss of the desired enantiomers.
According to one embodiment of the above method for isolation and purification, the S-enantiomer of the compound of formula (TV) is isolated from the R-enantiomer of the compound of formula (II).
According to another embodiment of the above method for isolation and purification, the S-enantiomer of the diol of foraiula (II) is isolated from the R-enantiomer of the acyl derivatitive of formula (TV).
The amount of water to be used is 1:2 to 1:100, preferably 1:5 to 1:50, as aratio between the compound of fomiula (E) and water. In addition, prior to the extraction with water, the reaction solvent may be evaporated to reduce the amount thereof or may be substituted with another organic solvent.
The acid which may be used in the above isolation and purification process is not particularly restricted, but for example, there may be mentioned, mineral acids such as hydrochloric acid, sulfuric acid and phosphoric acid; organic acids, in particular carboxyUc acids, represented by aUphatic carboxylic acids such as formic acid, acetic acid, propionic acid, n-butyric acid, iso-butyric acid, n-valeric acid, iso-valeric acid, pivaUc acid, n-caproic acid, iso-caproic acid, cyclopentanecarboxylic acid, cyclohexanecarboxylic acid, oxalic acid, malonic acid and tartaric acid; or aromatic carboxylic acids such as benzoic acid, p-chlorobenzoic acid, p-nitrobenzoic acid, p-methoxybenzoic acid, p-toluic acid, o-toluic acid, m-toluic acid, naphthoic acid, phthalic acid and telephtalic acid. Among them, organic acids are preferred, in particular carboxylic acids such as n-butyrate, iso-valerate, cyclohexanecarboxylic acid, pivalic acid, benzoic acid and o-toluic acid, and particularly preferred is pivalic acid. Needless to say, the above acids may be used singly or in a combination of two

or more species. The amount of the acids to be used is not particularly restricted, but the molar ratio of the sum of the compounds of formula (IV) and (II) and acid, is usually 1:1 to 1:5, and preferably 1:1 to 1:3.
Tlie acid used in the present isolation and purification step may be the same as the organic acid used in the acylation or deacylation reaction according to the invention or may be a different one.
As an organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, hydrocarbons such as hexane, heptane, benzene and toluene; ethers such as diethyl ether, tetrahydrofiiran, 1,4-dioxane, tert-butyl methyl ether and dimethoxyethane; ketones such as acetone, diethyl ketone and methyl ethyl ketone; esters such as methyl acetate, ethyl acetate, ethyl butyrate and ethyl benzoate; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane. Among them, aliphatic hydrocarbons, such as hexane and heptane and aromatic hyderocarbons, such as benzene and toluene are preferred. Most preferred are aromatic hydrocarbons and most preferred is toluene. The above solvents may be used singly or in a combination of two or more solvents.
Temperature of the isolation and purification step is preferably at 0 to 80 °C, and more preferably at 10 to 40 ""C and most preferred at 20 to 30 °C
Following, separation of the salts of the compounds of formula (EQ and (TV) according to the above method for isolation and purification, minor amounts of the compound of formula (IV) which is partly mixed in an aqueous layer of the salt of the compound of formula (II) with an acid, and the like can be efficiently removed by wasliing the aqueous layer with an organic solvent.
The salt of the diol of formula (II) may be used as an aqueous solution of the salt, and if needed, it may be used as a solution in another solvent or a concentrate obtained by concentration, solvent substitution or the like operation. Furthermore, it can be used as a crystal obtained by crystallization or the like operation. However, it is usual that the diol of formula (II) is be used as a free diol form obtained by the following operations: A mixture containing the diol of formula (11) and an organic solvent

and/or its concentrate is obtained by treating an aqueous layer with a general base such as sodium hydroxide or potassium hydroxide to control the pH of the aqueous layer to be at least 9, preferably at least 11, then extractmg the free amine form of the diol of formula (II) with an organic solvent, followed by washing and concentrating the extract. The chemical purity of the diol of formula (II) obtained by the series of isolation and purification method is usually at least 95%, preferably at least 97%, more preferably at least 99% and most preferably at least 99.5%.
On the other hand, the compound of formula (TV) obtained by the above operations may be washed with an aqueous phase in order to improve chemical purity of the product. The compound of formula (IV) may be obtained as a free amine form by treating the ammonium salt of the compound of foraiula (TV), obtained by the above operations, with a base. The chemical purity of the diol of formula (TV) obtained by the series of isolation and purification method is usually at least 95%, preferably at least 97%, more preferably at least 99% and most preferably at least 99.5%.
Tlie optical purity of the product obtained after separation of the enantiomers of formula (II) and (IV) as above, may be improved before fiirther processing. Improvement of the optical purity may be obtained by chromatography as described in WO 03/011278 or by crystallisation of diastereomeric esters or salts with optically active acids as described in US patent No. 4,943,590.
According to a further embodiment of the invention, the mixture of compound of formula (11) and compound of formula (TV) which is separated by the above method for isolation and purification has been prepared by the selective acylation and in another embodiment by selective deacylation according to the invention.
The invention also relates to another novel method for the separation of the R- or S-diol of formula (11) from the acyl derivative of formula (IV) of the other enantiomer whereby the desired compound may be isolated and purified.
According to this other method for isolation and purificationthe acyl derivative having the formula

wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line represents a double or a single bond, Z is a group -CH2-N(R'R') wherein R'and R' are Ci-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl group, W is 0 or S; and R^ is -Y-R^ wherin Y is a bond, 0, S or NH and R^ is C1-10-alkyl, C:.io-aIkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(CMo-alkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from CMQ-alkyl, C2-10-aIkenyl, C1-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci.io-alkylaminoand di-(Ci.io-allcyl)amino, or a salt thereof
and/or the diol of formula

wherein R, Hal, the dotted line and Z are as defined above, or a salt thereof, iso isolated from a mixture containing a compound of formula (IV) and a compound of formula (H), by a method comprising:
a) treating said mixture containing the compound of formula (IV) and the diol of formula (II) in a mixture of water, a protic organic solvent and an apolar organic solvent;
b) separating the aqueous phase containing the diol of formula (II), firom the organic phase to obtain an organic phase containing the compound of formula (IV);
optionally isolating the diol of formula (II) and/or (IV) as a base and optionally conversion of the compound of formula (II) and /or (IV) to a salt thereof.
Any of the above isolated phases (aqueous and organic) may additionally be washed one or more times with an organic or an aqueous solvent, respectively to improve the chemical purity of the product.
According to one embodiment of the above method for isolation and purification, the S-diol of the compound of foraiula (II) is separated firom the R-enantiomer of the compound of formula (IV).
According to another embodiment of the above method for isolation and purification, tlie S-enantiomer of the compound of fomiula (IV) is separated from the R-enantiomer of the compound of formula (II).
According to one embodiment of the invention R is halogen or cyano, preferably cyano in the method for isolation and purification above.
According to another embodiment of the invention Hal is fluoro in the method for isolation and purification above.
According to a preferred embodiment, the dotted line represents a single bond.

In another embodiment of the invention, Z is dimethylaminomethyl or a group that may be converted to a dimethylamiaomethyl group. Suitable Z is dimethylaminomethyl.
According to a preferred embodiment of the invention Hal is fluoro, Z is dimethylaminomethyl, the dotted line is a stQgle bond and R is cyano or iialogen, preferably cyano.
In a further embodiment of the mvention the compound of foraiula (IV) in the method for isolation and purification above, is a compound wherein Y is O, or S, preferably Y is O and tlie other substituents are as defined above. In a further embodiment of the invention the compound of formula (IV) in the method for isolation and purification above, is a compound, wherein Y*is S.
In a further embodiment of the invention the compound of formula (TV) in the method for isolation and purification above, is a compound wherein Y is a bond and the other substituents are as defined above.
In a further embodiment of the invention the compound of formula (TV) 'm the method for isolation and purification above, is a compound wherein Y is NH and the other substituents are as defined above.
In a preferred embodiment of the invention, the compound of formula (IV) in the method for isolation and purification above, is a compound wherein R^ is C1-6-alkyl, C2-6-alkenyl or Ci-e-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-6-alkoxy, Ci-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.6-alkyl)amino, more preferred R^ is Ci.4-alkyl, Cz^-allcenyl or Ci-i-allcynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-all
In anotlaer preferred embodiment of the invention, the compound of formula (IV) in the method for isolation and purification above, is a compound wherein R^ is as follows: C1-10-alkyl, C2-10-alkenyl or C2-10-aIkynyl all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, CMO-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-6-alkylamino and di-(Ci.io-alkyl)amino, more suitable R^ is Ci.io-alkyl, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected fi-om hydroxy, halogen, amino, nitro and cyaho, preferably R* is C1-10-alkyl, preferably unbranched d-io-alkyl and more preferred R^ is unbranched C4-io-alkyl
As an protic organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, alcohols such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanoI and tert-butanol. The above solvents may be used singly or in a combination of two or more species.
As an apolar organic solvent which may be used in the isolation and purification step, there may be mentioned, for example, hydrocarbons such 'as hexane, heptane, benzene and toluene; ethers such as diethyl ether, tert-butyl methyl ether and dimethoxyethane; halogenated hydrocarbons such as methylene chloride, chloroform and 1,1,1-trichloroethane. Among them, preferred are hydrocarbons such as hexane, heptane, benzene and toluene, and more preferred is heptane. The above solvents may be used singly or in a combination of two or more species.
According to a specific embodiment of the invention, the mixture of the compound of fonnula (TV) and the diol of formula (II) used in the above method for isolation and purification has been prepared by enzymatic acylation according to the invention and in another embodiment by enzymatic deacylation according to the invention.
The optical purity of the product obtained after separation of the enantiomers of fonnula (II) and (IV) as above, may have to be improved before further processing. Improvement of the optical purity may be obtained by chromatography as described in WO 03/006449 or by crystallisation of diastereomeric esters or salts with optically active acids as described in US patent No. 4.943.590.

The present invention also relates to a process for the preparation of escitalopram having the formula

or a phannaceutically acceptable salt thereof comprising preparation of the S-enantiomerofadiolhavingthe formula

wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond, Z is a dimethylaminomethyl group or a
group which may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof,
or the S-enantiomer of an acylated did having the formula

wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R^ wherein R^ is Ci.io-alkyI, C1-10-alkenyl or C1-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom CI-IQ-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(Ci.i{ralkyI)amino, aryl, aryloxy^ aiylthio and heteroaryl, or R* is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2.io-alkenyl, C2.io-alkynyl, Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkyianiino and di-(Ci.io-alkyl)amino and Y is a bond, 0, S or NH or a salt thereof, in one embodiment by the method for selective enzymatic acylation according to the invention as defined in any of the embodiments above audr in another embodiment by the method for selective enzymatic deacylation according to the invention as defined in any of the embodiments above, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and ring closure under basic conditions of the S-enantiomer of formula (Us) or (IVs) or a labile ester derivative thereof to form a compound of foraiula

followed by, in either order, if R is not cyano conversion of the group R to a cyano group, if the dotted line represents a double bond reduction to a single bond, if Z is not dimethyiaminometfayl conversion of the group Z to a dimethylaminomethyl group and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolation of escitalopram or a phannaceutically acceptable salt thereof
According to one embodiment of the invention, the S-enantiomer of formula (lis) above or the S-eantiomer of formula (TVs) used for the preparation of escitalopram is separated from the R-enantiomer of formula (IV) and (11) respectively, before ringclosure.
According to one embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (II) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according one of the above novel methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above novel methods for isolation and purification.
According to still another embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (U) and the opposite enantiomer of the compound of formula (IV) obtained by enzymatic deacylarion has been separated from each other by the isolation and purification process according one of the above novel methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above novel methods for isolation and purification.

According to another embodiment of the invention, the S-enantiomer of foraiula (lis) above or the S-enantiomer of formula (TVs) used for the preparation of escitalopram is not separated from the R-enantiomer of foraiula (IV) and (IT) respectively, before ringclosure.
As mentioned above, the group R means cyano or any other group which may be converted to a cyano group.
Groups v^hich may be converted to a cyano group include halogen such as chloro, bromo, iodo or fluoro, preferably chloro or bromo.
Other groups which may be converted to cyano include CF3-.(CF2)n-SO2-0-, wherein n is 0-8, -OH, -CHO, -CH2OH, -CH2NH2, -CH2NO2, -CH2CI, -CHiBr, -CH3, -NHR^ -CHNOH, -COOR^ -CONRV wherein R^ is hydrogen or C1-6 alkylcarbonyl, and R^ and R^ are selected from hydrogen, optionally substituted C1-6 atkyl, aryl-C1-6 alkyl or aryl and, a group of fomiula

wherein Z is O or S; R^ - R^ are each independently selected from hydrogen and Ci-6 alkyl or R^ and R^ together form a C2.5 alkylene chain tiiereby forming a spiro ring; R^° is selected from hydrogen and C1-6 aUcyl, R^^ is selected from hydrogen, Ci,6 alkyl, a caxboxy group or a precursor group therefore, or R^ and R together forai a C2-5 alkylene chain thereby forming a spire ring.
When R is halogen, in particular bromo or chloro, conversion to a cyano may be carried out as described in US 4,136,193, WO 00/13648, WO 00/11926 and WO 01/02383.
According to US 4,136,193 conversion of a bromo group to a cyano group, is carried out by reaction with CuCN.

wo 00/13648 and WO 00/11926 describe tlie conversion of a halogen or a triflate group to a cyano group by cyanation with a cyanide source in presence of a Pd or Ni catalyst.
Other processes for the conversion of a bromo compound to the corresponding cyano derivative involve reaction of the bromo compound with magnesium to form a Grignard reagent, followed by reaction with a formamide to form an aldehyde. The aldehyde is converted to an oxime or a hydrazone which is converted to a cyano group by dehydration and oxidation, respectively.
Alternatively, the bromo compound is reacted with magnesium to form a Grignard reagent,, followed by reaction with a compound containing a CN group bound to a leaving group.
A detailed description of the above two procedures may be found in WO 01/02383.
Compounds wherein the group R is -CHO, may be converted to the corresponding compounds wherein R is cyano by methods analogous to those described in WO 99/00210.
Compounds wherein the group R is NHR^^, wherein R^^ is hydrogen or alkylcarbonyl, may be converted to the corresponding compounds wherein the group R is cyano by methods analogous to those described in WO 98/19512.
Compoimds wherein R is a -COOR^ group, may be converted to the corresponding :ompound wherein R is cyano by conversion to the amide via the corresponding acid :hloride or an ester thereof followed by dehydration of the amide. WO 01/68632.
Alternatively, a compound where R is -COOH may be reacted with chlorosulfonyl isocyanate in order to form the nitrile; cr treated with a dehydrating agent and a sulfonamide as described in WO 00/44738.
Tompounds wherein the group R is -CONR^'^R^'^, wherein R^' and R^"^ are selected from hydrogen optionally substituted allcyl, aralkyl or aryl may be converted to the

corresponding cyano compound by methods analogous to those described in WO 98/00081 and W098/1951L
Compounds wherein the group R is a group of formula (VIE) may be converted to the corresponding cyano compound by methods analogous to those described in WO 00/23431.
Compounds wherein R is OH, -CH2OH, -CH2NH2, -CH2NO2, -CH2CI, -CHaBr, -CH3 or any of the groups above, may be converted to the corresponding cyano compounds by methods analogous to those described in WO 01/68632.
Racemic compounds of formula (II) may be prepared by the methods described in the above mentioned patents or by the alkylation method described in US patent No. 4.136.193 or the double grignard reaction described in EP 171 943 or by analogous methods. Racemic compounds of formula (TV) may be prepared from racemic compounds of formula (II) by non-selective acylation using anhydrides, esters, carbonates, isocyanates or isothiocyanates as defined by formulas (IQa), (Hlb), (Die), R^-N=C=0 andR^-N=C=S above.
In some cases the racemic compound of formula (E) may be available in the fomi of an acid addition salt, such as the sulphate salt, and in this case a free base of the compound of formula (II) may be obtained by treating the salt with a base in a mixture or water and an organic solvent, to transfer the compound of formula (II) into the organic phase.
Preferably, R is cyano in the compounds of romula (H), (lis), (Ilr), (IV) (IVs) (IVr) and (V). If R is not cyano, conversion of the group R to a cyano group is suitably carried out after ringclosure to form a compound of formula (V).
Preferably, Hal is fluoro in the compounds of romula (H), (lis), (Er), (IV) (IVs) (IVr) md (V).If Hal is not fluoro, conversion of the group Hal to a fluoro is suitably carried Dut after ringclosure to form a compound of formula (V). A procedure for carrying out :his conversion is described in Speciality Chemicals Magazine, April 2003, page 36-58.

Z groups which may be converted to dimethylaminomethyl are groups such as -CH2-L, -CH2-NO2, -MgHal, cyano, aldehyde, -CH2-0-Pg, -CHz-NPgjPga, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, -CO-N(CH3)2, -CH(A^R')(A'R^0, -(A'R")(A'R')(A^R'^, -COOR'VCH2-CO-NH2, -CH=CH-R'« or-CONHR", wherein Pg is a protection group for an alcohol group, Pgi and Pg^ are protection groups for an amino group, R^^ and R'^ are independently selected from C1-6 alkyl, C2-6 alkenyl, C1-6 alkynyl and optionally alkyl substituted aryl or aralkyl groups or R^^ and R'^ together form a chain of 2 to 4 carbon atoms, each of R*'* - R'^ are independently selected from C1-6 alkyl, Cz^ alkenyl, 2i^ alkynyl and optionally C1-6 alkyl substituted aiyl or aryl-C1-6; alkyl, R'^ is hydrogen or methyl and A', A^ and A^ are selected form O and S; L is a leaving group, such as halogen or -O-SO2-A wherein A is C1-6 alkyl, Cz^ alkenyl, C2-6 alkynyl or optionally C1-6 allcyl substituted aryl or aryl-Ci-6 alkyl.
In one embodiment of the invention Z is dimethylaminomethyl, -CH2-L, -CH2-NPg,Pg2, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, -CO-N(CH3)2, aldehyde or -COOR'"^ in the compounds of formula (II), (Us), (Ilr), (IV) (IVs) (IVr) and (V).
Compounds wherein Z is -CH2-0-Pg may be converted to the correspondiag compounds wherein Z is dimethylaminomethyl by removal of the protection group to form the corresponding alcohol and thereafter conversion of the alcohol group to a feasible leaving group and reaction of the resulting compound with
a) dimetlaylamine or a salt thereof,
b) methylamine followed by methylation or reductive amination, or
c) with an azide followed by reduction to form the corresponduig amine and tlaereafter methylation or reductive amination, as described in WO 01/43525, WO 01/51478 or WO 01/68631 or by analogous methods.
Compounds wherein Z is -CH2-L, wherein L is a leaving group, may be converted fo ' a dimethylaminomethyl group in the same manner.
Compounds wherein Z is -CO-N(CH3)2 and -CO-NHR'^ wherein R'^ is hydrogen or methyl, may be converted to the corresponding compounds wherein Z is

dimethylaminomethyl by reduction of the amide and if a primary or secondary amine . is formed, followed by methylation or reductive amination to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods.
Compounds wherein Z is -CH2-NMe(PgO or -CH2-N(Pgl)(Pg2) may be converted to the corresponding compound wherein Z is dimethylaminomethyl by removal of the protection groups, i.e. to obtain compounds wherein Z is -CH2-NH2 or -CHi-NMe and thereafter methylation of the amino group or reductive amination to form a dimethyaminomethyl grot^ as described in WO 01/43525 or WO 01/68631 or by analogous methods.
Compounds wherein Z is -CH(A^R^^)(A^R*^) may be converted to the corresponding compounds wherein Z is dimethylaminomethyl by deprotection to form an aldehyde (compounds where Z is -CH2-CA^H) followed reductive amination with dimethylamine to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods.
Compounds wherein Z is -C(A^R^^)(A^R^^)(A^R^^ may be converted to the
corresponding compounds wherein Z is dimethylaminomethyl
i) by hydrolysis to fomi a carboxyhc acid or an ester thereof and thereafter
reduction to forai an alcohol and thereafter conversion of the alcohol group to a
feasible leaving group, followed by replacement of the leaving group with a
dimethylamino group as described above, or
ii) conversion of the carboxyhc acid or an ester thereof to an amide by reaction
with an amine of NH(Me)2, NHaMe, NH3 or a salt thereof foUowed by, in either
order, reduction of the amide and if necessary methylation or reductive amination to
form a dimethylamino group as described in WO 01/68631 or by analogous methods.
Compounds wherem Z is -COOR^^ may be converted to the corresponding compounds wherein Z is dimethylaminomethyl as described^above, starting with the carboxyhc acid ester.
Compounds wherein Z is -CH2-CONH2 may be converted to the corresponding compound wherein Z is dimethylaminomethyl by treatment with hypohalide to form a

primary amine followed by methylation of the free amino group or reductive amination to form a dimethylaminomethyl group as described in WO 01/43525 or WO 01/68631 or by analogous methods.
Compounds wherein Z is -CH=CHR^^ may be converted to the corresponding compound wherein Z is dimethylaminomethyl by oxidation to fomi an aldehyde which^may be converted to dimethylaminomethyl by reductive amination as described in WO 01/43525 or WO 01/68631 or by analogous methods.
Compounds wherein Z is cyano or-CH2-N02 may be converted to the corresponding compound wherein Z is dimethylaminomethyl by reduction followed by methylation or reductive amination of the free of the amino group formed to form dimethylaminomethyl as described in WO 01/68629 or by analogous methods.
Compounds wherein Z is -MgHal may be converted to the corresponding compound wherein Z is dimethylaminomethyl by reaction with dimethylaminomethyl alkyl ether of formula (CH3)2NCH20-alkyl as described in WO 01/68629 or by analogous
methods.
Preferably, Z is dimethylaminomethyl. If Z is not dimethylaminomethyl, conversion of Z to a dimethylaminomethyl group is suitably carried out after ringclosiure.
Suitably, Z is dimethylaminomethyl, -CH2-L, -CH2-NPgiPg2, -CH2-NMePgi, -CH2-NHCH3, -CH2-NH2, aldehyde, -CO-N(CH3)2, -COOR*^ when the dotted line represents a bond.
Compoirnds of formula (II) or (IV) wherein Z is dimethylaminomethyl, CH2-NPgiPg2 ,-CH2-NMePgi, -CH2-NHCH3, or -CH2-NH2, in particular compounds wherein Z is dimethylaminomethyl, -CH2-NHCH3, or -CH2-NH2, are particularly well suited for the metliod of purification isolation according to the invention, wherein the compounds of formula (II) and (IV) are separated.
In the compounds of romula (11), (lis), (Hi), (IV) (TVs) (TVr) and (V), the dotted line is preferably a single bond. Compounds wherein the dotted line represents a double

bond may be converted to the corresponding compound wherein the dotted line is a single bond by the methods described in WO 01/68630, Preferably the reduction is carried out after ringclosure.
Enantioselective ring-closure of the compounds of formula (IVs) or (lis) to form the compounds of fomiula (V) may suitably be carried out by treatment of a labile ester derivative of the compound with a base such as KOC(CH3)3 and other alkoxides, N^ and other hydrides, triethylamine, ethyldiisopropylamine or pyridine, at low temperatures in an inert organic solvent, such as tctrahydrofuran, toluene, DMSO, DMF, t'butyl methyl ether, dimethoxyethane, dimethoxymethane, dioxane, acetonitrile or dichloromethane. This process is described in US patent No. 4,943,590.
Ringclosure of the compound of formula (lis), is suitably carried out by treatment
with a base as described above in presence of an agent capable of forming a labile
group with the primary alcohol of the diol, such as methanesulfonyloxy, /?-
toluenesulfonyloxy, 10-camphorsuIfonyloxy, trifluoroacetyloxy and
triiluoromethanesulfonyloxy and halogen.
In some cases, it may be advantageous to exchange the -W-R"^ group m the compound
of formula (TVs), for a more labile group before ringclosure is carried out. Such labile
groups (leaving groups) could typically be a group selected from methanesulfonyloxy,
/7-toluenesulfonyloxy, 10-camphorsulfonyloxy, trifluoroacetyloxy and
trifluoromethanesulfonyloxy or halogen.
Typically, the compound of formula (IVs) is subjected to hydrolysis to form the compound of formula (lis) with aquesous base, such as NaOH, KOH or LiOH in water or alcohol or a mixture thereof and then reacted with an activated leaving group, such as for example mesylchloride or tosylchloride in an organic solvent in the presence of an organic base.
The optical purity of the escitalopram product may have to be improved after ringclosure. Improvement of the optical purity may be obtained by chromatography

on a chiral stationary phase or by crystallisation of racemic citalopram base or a salt ^ thereof according to the methods described in WO 03/000672.
The R-enantiomer of the compounds of formula (E) and (IV) obtained according to the invention may be used to prepare racemic citalopram and escitalopram by ring closure in acidic environment according to the method described in WO 03/000672. Suitable acids for caxiying out acidic ring closure are raiQeraLacid, a carboxylic acid, a sulfonic acid or sulfonic acid derivative, more suitable H2SO4 orH3P04.
Thus in another embodiment, the invention relates to a process for the preparation of racemic citalopram and /or escitalopram or a pharmaceutically acceptable salt thereof comprising preparation of the R-enautiomer of a diol having the formula

wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents an optinal bond and Z is a dimethylaminomethyl group or a group which may be converted to a dimedaylaminomethyl group and Hal is halogen, or a salt thereof,
or the R-enantiomer of an acylated diol having the formula

wherein R, Z, the dotted line and Hal is as defined above, W is 0 or S, and R is -Y-R\ wherein R^ is CMo-alkyl, C2-10-alkenyl or C1-10-alkynyi all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy,' halogen, amino, nitro, cyano, Ci.io-alkylamino, di-(CMo-alkyl)amino, aryi, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaiyl groups may optionally be substituted one or more times with substituents selected from Ci.io-alkyl, Cj-io-alkenyl, C2-10-alkynyl, Ci.io-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, CMo-alkylamino and'di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt diereof, in one embodiment by the method for selective enzymatic acylation according to the invention as defined in any of the embodiments above and in another embodiment by the method for selective enzymatic deacylation according to the invention as defined in any of the embodiments above, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and ringclosure under acidic conditions of the R-enantiomer of fonnula (Ur) or (TVs) to form a mixture of the compound of formula

wherin Hal, Z, R and the dotted line are as defined above, and a minor amount of the corresponding R-enantiomer, followed by, in either order; if R is not cyano conversion of the group R to a cyano group, if the dotted line represents a double bond reduction to a single bond, if Z is not dimethylaminomediyl conversion of the group Z to a dunethylaminomethyl group and if Hal is not fluoro, conversion of Hal * to a fluoro group, followed by isolation of escitalopram and/ or racemic citalopram or a pharmaceutically acceptable salt thereof, by precipitation of racemic citalopram free base or a salt thereof and recovery of escitalopram form the mother liquor of the precipitation.
Suitably, the above method for conversion of R-diol to escitalopram, is used for the preparation of escitalopram.
The conversion of of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and conversion of Hal to a fluoro group may be carried out as described above.
According to one embodiment of the above method for the preparation of escitalopram the mixture of the R- or S-enantiomer of a compound of formula (11) and tlie opposite enantiomer of the compound of formula (IV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according t)ne of the above methods for isolation and purification. According to another embodiment, the mixture has been separated by the other of the above methods for isolation and purification.

According to still another raibodiment of the above method for the pr^aration of escitalopram the mixture of the R- or S-CTiantiomer of a compound of foraiula (II) and the opposite enantiomCT of the compound of formula (IV) obrtained by enzymatic deacylation has been separated from each other by the isolation and purification process according one of the above methods for isolation and purification- According to another embodiment, the mixture has been separated by the other of the above methods for isolation and purification.
According to another embodiment of the invention, the S-enantiomer of formula (Or) above or the S-enantiomer of fomiula (TVr) used for the preparation of escitalopram is not separated forai the R-enantiomer of formula (IV) and (IT) respectively, before ringclosure.
The optical piirity of the escitalopram product may have to be improved after ringclosure. Improvement of the optical purity may be obtained by chromatography on a chiral stationary phase or by crystallisation of racemic citalopram base or a salt thereof according to the methods described in WO 03/000672.
As used herein, the term C1-10-alkyl refers to a branched or imbranched alkyl group having from one to ten carbon atoms inclxisive, such as methyl, ethyl, l-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl, 2-methyl-l-propyl, pentyl, hexyl and heptyl. Ci-6-alkyl refers to a branched or unbranched alkyl group having from one to six carbon atoms inclusive, such as methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl, 2-methyH-propyl, pentyl and hexyL CM-alkyl refers to a branched or unbranched alkyl group having from one to four carbon atoms inclusive, such as methyl ethyl, l-propyl, 2-propyl, 1-butyl, 2-butyl, 2-methyl-2-propyl and 2-methyl-1-propyl. C1-3-alkyl refers to a branched or unbranched alkyl group having from one to three carbon atoms inclusive, such as methyl, ethyl, l-propyl, 2-propyl.
Similarly, C1-10-alkenyl and C2-10-alkynyl designate branched or unbranched alkeuyl and alkynyl groups, respectively, having from two to ten carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl. C2^-allcenyI and C2-^-allcynyl designate branched or unbranched alkenyl and alkynyl groups, respectively, having from two to six carbon

atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl Cz^^-alkenyl and C2^-alkynyI designate branched or unbranched alkenyl and alkynyl groups, respectively, having from tv^'o to four carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, butenyl, ethynyl, propynyl and butynyl. C2-3-alkenyl and C2-3-aIkynyl designate branched or unbranched alkenyl and alkynyl groups, respectively, having from two to three carbon atoms, including one double bond and one triple bond respectively, such as ethenyl, propenyl, ethynyl and propynyl.
The terms CMO alkoxy, CMO alkylthio, CMO alkylamino and di-(C1-10-alkyl)ainino etc. designate such groups in which the alkyl group is CMO alkyLas defined above. The terms C\^ alkoxy, CM alkylthio, C1-6 alkylamino and di-(C1-6-alkyl)amino etc. designate such groups in which the alkyl group is Ci-6 alkyl as defined above. The terms CM alkoxy. CM alkylthio. CM alkylamino and di-(CM-alkyI)amino etc. designate such groups in which the alkyl group is CM alkyl as defined above. The terms C1.3 allcoxy, C1-3 alkylthio, C1.3 alkylamino and di-(C]-3"aIkyl)aniino etc. designate such groups in which the alkyl group is C1-3 alkyl as defined above.
Halogen means fluoro, chloro, bromo or iodo.
The term aryl refers to a mono or bicycUc carbocyclic aromatic group, such as phenyl, or naphthyl, in particular phenyl.
Aryloxy, arylthio refers to such a group wherein aryl is as defined above.
The term heteroaryl refers to a 5 or 6 membered monocyclic heteroaromatic group or a bicyclic heteroaromatic group. Suitable the heteroaryl group contains 1-3 heteroatoms selected from 0, S and N.
R° and R' may together form a chain of 3 to 5 carbon atoms, thus forming an anhydride.

When Z is -CHrNCRTl") wherein R'and R" connected to each other to form a cyclic structure including the N-atom to which they are attached, the cyclic structure form groups, such as groups include piperidin, pyrrohdin, moipholinyl and piperazinyl
HOBt means hydroxybenzotriazol and pj^ means pentafluorophenoL
i
Experimentals
In the following examples % conversion and optical purity were measured and calculated as described below:
HPLC analysis condition (for conversion rate) used for examples 1-28:
Column: YMC-Pack ODS-A 0.46 cm I.D. x 25 cm (made by YMC Co., LTD.) Eluent: 25 mM Phosphate buffer / Acetonitrile = 60/40 Flow rate: LOml/min Temperature: 40°C Detector wavelength: 237 nm
HPLC analysis condition (for conversion rate) used for examples 29-42:
Column: A Lichrospher RP-8 column, 250 x 4 mm (5 ^un particle size)
Eluent: Bufifered MeOH/water prepared as follows: 1,1 ml EtsN added to 150 ml
water, 10% H3P04(aq) is added to pH=7 and water is added to a total of 200 ml. The
mixture is added to 1,8 L MeOH.
Temperature; 35^C Flow rate: 1 mL/min Pressure: 16,0 MPa Detection: UV 254 nm 'Injection volume: 10 microL
Conversion rate (%) = P/(S+P) x 100, (P: amount of product, S: amount of residual substrate).

HPLC analysis condition (for optical purity) used for examples 1-28:
Column: Chiralpak AD 0.46 cm I.D. x 25 cm (made by DAICEL CHEMICAL
INDUSTRIES, LTD.)
Eluent: Heptane / methanol / ethanol / diethylamine = 85 / 7.5 / 7.5 / 0.1
Flow rate: l.Oml/min
Temperature: 30°C
Detector wavelength: 240 nm
Conversion rate (%) = P/(S+P) x 100, (P: amount of product, S: amount of residual
substrate)
Super critical fluid chromatogeaphy. Analysis condition (for optical purity) used for examples 29-42:
Column: Daicel AD column with the dimensions 250 x 4,6 mm (5 |j.m particle size) Mobile phase: Carbon dioxide
Modifier: Methanol with diethylamine (0.5%) and trifluoroacetic acid (0.5%). Modifier gradient: 1-2% in 4 minutes
2-4% in 4 minutes
4-8% in 4 minutes
8-16%in4minutes
16-32% in 4 minutes
32-45% in 1,62 minutes
Temperature: Ambient temperature Flow rate: 2 mL/min Pressure: 20 mPa Detection: UV 230 nm and 254 nm Injection volume: lOmicroL
Optical purity (% ee) = (A-B) / (A+B) x 100, (A and B represent corresponding stereo isomer, A>B)
E-value - hi ((1-c/lOO) x (1-Es/lOO)) / In ((1-c/lOO) x (l+Es/lOO))

(c: convertion ratio, Es: optical purity of residual substrate) Example 1
(S)-4-[4-Dimethylainino-l-(4'-fluorophenyl)-l'hydroxybutyl1-3-hydroxymethyl-benzonitrile
Water (200.7 g) and toluene (440.8 g) was added to (±)-4-[4-dimethylamino-l-(4-
fluorophenyl>l-hydroxybutyl]-3-hydroxymethylben2omtrile, 1/2 sulfuric acid salt
(100.4 g, 0.257 mol), then stirred for 15 minutes and the temperature thereof was
raised up to 60**C. Subsequently, 30% NaOH (34.5 g, 0,259 mol) was added dropwise
over 5 minutes until pH of aqueous layer reached 11.4, and stirred for 30 minutes.
After stinring, the mixture was allowed to stand for 5 minutes, and separated. The
aqueous layer was discarded, and thus-obtained organic layer was washed with 220,0
g of water, and further, concentrated under reduced pressi3xe at 60°C to remove away
the remaining moisture. The concentration was adjusted by addition of toluene, to
thereby obtain 583.1 g of a solution of 4-[4-dimethylamino-l-(4'-fluorophenyl)-
l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene (content of pure 4-[4-
dimethylamino-l-(4'--fluorophenyl)-l-hydroxybutyi]-3-hydiDxymethylbenzonitrile:
87.6 g, yield: 99.7%, 0.256 mol). Then, 52.5 g of toluene was poured onto 17.5 g of
an immobilized enzyme (Novozym 435), then 583.1 g of the solution of 4-[4-
dimethylamino-1 -(4'-fluorophenyI)-1 -hydroxybutyl]-3-hydroxymethylbenzonitrile in
toluene was added, and the temperature of the mixture was controlled to 40°C.
Subsequently, a mixed solution containing pivalic acid (28.7 g, 0.281 mol), vinyl
butyrate (29.2 gm 0,256 mol) and toluene (28.8 g) was added into the above mixture
over 20 minutes, and the mixture was stirred at 40°C for 12 hours under a slight flow
of nitrogen. After cooling the mixture over 2 hours so as to be 20°C, the mixture was
stirred for 1 hour at the same temperature, and stirring was stopped. The enzyme was
filtrated, and then the enzyme was washed twice with 105 g of toluene, and these
• - • -
toluene layers were combined. The toluene layer contained 583.1 g of a solution of 4-
[4-dimethylamino-l-(4'-.fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtriIe
in toluene (content of pure (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l-
hydroxybutyl]-3-hydroxymethyIbenzonitrile: 36.0 g, yield through the enzymatic
resolution process: 41.1%, 0.105 mol).

A mixed solution containii^ the supernatant and washing fluid was extracted three times wifli water (438 g, 263 g x 2), and 1005.4 g of the obtained aqueous layer was washed with 79 g of toluene for five times. As a result, it was obtained 985,2 g of an aqueous solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophaiyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt {content of pure (S)-4-[4-dimethyfaniino-l-(4'-fluorophenyI)-l-hydroxybutyl]-3-hydroxymethyn3enzoni 32,6 g, yield (through the process from the extraction with water to washing with tohiene): 90.6 %, 0.0952 mol}. To thus-obtained 985.2 g of an aqueous solution of (S)-4-[4-dimethylamino-l-{4*-fliiorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was added 218^ g of toluene. Subsequently, 30% NaOH (13.3 g, 0.0998 mol) was slowly added to the mixture with stirring, until pH of aqueous layer reached 11.9, and the mixture was kept stirred for 30 minutes. After stilting, the mixture was allowed to stand 30 minutes and the toluene layer was separated, and the remaining aqueous layer was re-extracted with 142.5 g of toluene. The combined toluene layer was washed twice with 169.0 g of water, concentrated at 60**jC under reduced pressure, to thereby obtain 64.0 g of a solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzomtrile in toluene. Said toluene solution contained 32.0 g (0.0935 mol) of (S)-4.[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile. Overall yield was 36.4%. The optical purity determined by HPLC was 98.7 % ee, and chemical purity was 99.9 area %, {(R)-5-cyano-2-[dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 0.04 area %.
In the above system, die above "chemical purity" is represented by the following
equation.
(Chemical purity) = [Area value of 4-[4-dimethylainino-l-(4'-fluorophenyl)-l-
hydroxybutyl]-3-hydroxymethylben2onitrile] / [(Area value of all detected
compounds) - (Area value of toluene) - (Area value derived from the system)] x 100
(area %)
Example 2

A test tube equipped with a stopper was charged witti 50 mg ( 0.146 m mol) of (±)-4-^ [4-dimethylaimno-H4'-fIuorophenyl)-l-hydroxybutyl]-3-hyd^^ 25 mg of various kinds of Upases, 33 mg ( 0.29 m mol) of vinyl butyrate and 1 ml of toluene, and stirred at 40**C for 16 hours. For the substrate with the optical purity of at least 50% ee, E-values were calculated. The results are shown in Table 1.

Example 3
A test tube eqiiipped with a stopper was charged with 10 to 100 mg (0.029 to 0.29 m mol) of (±)-4-[4-dimethyIamino-1 -{4'-fluorophenyl)-1 -hydroxybutyl]-3" hydroxymethylbenzonitrile, 10 to 100 mg of Novozym 435 (product of Novozymes, E/S ratio = 1.0), 33 to 333 mg (10 eq.) of vinyl butyrate and 1 ml of toluene, and stirred at 30°C for 16 hours. The results are shown in Table 2.

Example 4
A test tube equipped with a stopper was charged with 100 mg (0.292 m mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl)-3-hydroxymethyl-

benzonitrile, 50 mg of Novozym 435 product of Novozymes), 333 mg ( 2.92 m mol) of vinyl butyrate, 0.292 m mol of various kinds of additives and 1 ml of toluene, and stirred at the temperature of 40®C for 16 hours. The results are shown in Table 3.

Example 5
In a 200-ml four-necked flask equipped with a stirrer and a thermometer, 1.5 g (4.38 m mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.45 g of Novozym 435 (product of Novozymes), 0.347 g ( 4.38 m mol) of pyridine, 4.38 m mol of various kinds of acids and 1.00 g (8.76 m mol) of vinyl butyrate were added, and stirred at 40*'C for between 16 and 21.5 hours. The result is shown in Table 4.

Example 6
In a 200-ml four-necked flask equipped with a stirrer and a thermometer, 3.0 g (8.76 m mol) of (±>4-[4-dimethylaraino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.9 g of Novozym 435 (product of Novozymes), 0.693 g ( 8.76 m mol) of pyridine, 8.76 m mol of various kinds of acids and 2.00 g (17.52 m mol) of vinyl butyrate were added, and stirred at 40^C for between 15.5 and 16.5 hours. The results are shown in Table 5.

Example 7
In a 200-inl four-necked flask equipped with a stiirer and a thennometer, 4,5 g (13.1 m mol) of (±)-4-[4-dimethylammo-l-(4'-fluorophenyl)-l-liydroxybutyl]-3-hydroxymethylbenzonitrile was dissolved in 30 ml of toluene, then 0.90 g of Novozym 435 (product of Novozymes), 2.68 g (26.2 m mol) of pivalic acid and 0.900 to 1.500 g (0.6 to 1.0 eq.) of vinyl butyrate were added, and stirred at 40T for 24 hours. The restilts are shown in Table 6.

Example 8
In a 500-inl separable flask equipped with a stirrer and a thermometer, 20.0 g (58.4 mmol) of (±)-4-(4-dimethylamino-l-(4'-fluorophenyl)-l-hyd[roxybutyl)-3-hydroxymethylbenzonitrile was dissolved in 144 ml of toluene, then pivalic acid (1.1 to 3 eq.), 4.0 g of Novozym 435 (product of Novozymes) and 6.67 g (58,4 m mol) of vinyl butyrate were added, and stirred at 40*'C for 20 to 24 hours in a flow of nitrogen (5 ml/min). The results are shown in Table 7.

Example 9
A test tube equipped with a stopper was charged with 10 mg ( 0.029 m mol) of (i)-4-
[4-dimethylainino-l-{4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbe^^ ,
10 mg of Novozym 435 (product of Novozymes), 0.29 m mol of various kinds of acyl donor and 1 ml of diisopropyl ether, and stirred at 30**C for 16 hours. For the substrate with the optical purity of at least 30% ee, E-values were calculated. The results are shown in Table 8.

Example 10
A test tube equipped with a stopper was charged with 10 mg (0.029 m mol) of (i)-4-[4-dimethylaminO' 1 -(4'-fluorophenyl)-1 -hydroxybutyl]-3 -hydroxymethylbeozonitrile,

iKj nxg oi iNovozym 4i3 (product of Novozymes), 33 mg ( 0^9 m mol) of vinyl butyrate and 1 ml of various kinds of solvents, and stirred at 30°C for 16 hours. After the reaction, converted ratios and optical purities were analyzed and E-values were calculated. The results are shown in Table 9.

Example 11
A 200-nil four-necked flask equipped with stirrer and thermometer was charged with 0.9 to 1.35 g (E/S ratios are 0.2 to 0.3) of Novozym 435 (product of Novozym). A solution of 4.5 g (13.1 mmol) of (±)-4-[4-dimethyIamino-l-(4'-fluorophenyI)-l-hydroxybutyI]-3-hydroxymethylben2onitrile in 30 ml of toluene, 2.01 g of pivaUc acid (19.7 m mol), and 1.50 g (13.1 m mol) of vinyl butyrate was added into the flask.

and stirred at between 35 and 45*'C for 21 hours. After the reaction, reaction fluid was separated by decantation, and the remaining enzyme was washed with 30 ml of toluene and recovered. The same operation as described above except for using tiie recovered enzyme was repeated four times to cany out the recycle reaction Enzyme activities were calculated according to the following equation. The results are shown in Table 1 a
Enzyme activity (U/g) = (Product amount produced,per minute (fimol'min)) / (Enzyme weight (g))

Example 12
(S)-4-[4-DimethylaminO'l-(4'-fluorophenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt
A four-necked flask equipped with stirrer and thermometer was charged with 66.6 g of a solution of 10,0 g (0.029 mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and the temperature thereof was controlled so as to be 40*^0. Then, 2.27 g (0.029 mol) of pyridine, 2.98 g (0.029 mol) of pivalic acid, 3.35 g (0.029 mol) of vinyl butyrate, and 2.0 g of an immobilized enz3ane(Novozym 435) was added into the above mixture. The reaction mixture was . stirred at 40°CfoT 15 hours under a sUght flow of nitrogen and stirring was stopped. The enzjnne was filtrated from the reaction mixture with Kiriyama funnel, and the enzyme was washed with 21.6 g of toluene. As a result, it was obtained (S)- 4-[4-

dimethylamino-l-(4'-fluorophenyl)-l-hydioxybutyl]-3-hydroxy^
(3,6 g, O.OI 1 mol, yield: 36.0%, optical purity: 98.5 %ee), pivalic acid salt
The combined toluene layer was washed three times with water (50 ml, 30 ml, 30 ml)
at 2(fC, and an aqueous layer containing (S)"4-[4-dimethyiamino-l-(4'-flnorophenyl)-
l-hydroxybutyl]-3-hydroxymethyIbenzonitrile (3.5 g, 0.011 mol, extraction yield:
95%), pivalic acid salt was obtained.
Example 13
(S>4"[4-Dimethylamino--l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyl-beazonitrile pivalic acid salt
» • ...
A foxnr-necked flask equipped with stirrer and thermometer was charged with 66.7 g
of a solution of 20.0 g (0.058 mol) of (±)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l"
hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and the temperature thereof
was controlled so as to be 40°C. Then, 5.93 g (0.058 mol) of pivalic acid, 6.62 g
(0.058 mol) of vinyl butyrate, and 4.1 g of an immobilized enzyme (Novozym 435)
was added into the above mixture. The reaction mixture was stirred at 40'*C for 18
hours under a slight flow of nitrogen and stirring was stopped The enzyme was
filtrated from the reaction mixture with Kiriyama funnel, and the enzyme was washed
with 41.3 g of toluene. As a result, 6.7 g (0,020 mol) of (S)-4-[4-dimethylamino-l-
(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was
obtained iu the toluene solution, with a yield of 33.5% and an optical purity of 99.7
%ee.
Example 14
(S)-4-[4-Dimethylamino-l- (4'-fluorophenyl)'l ■hydroxybutyll-3-hydroxymetfayl'• benzonitrile cyclohexanecarfaoxylic acid salt
A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 25.0 g (0.073 mol) of (±)-4-[4-dimethyIamino-l-(4•-fluorophenyI)-l-hydroxybutyl]-3-hyd^oxymethylbenzomtrile in toluene, and the temperature thereof was controlled so as to be 40°C. Then, 5.77 g (0.073 mol) of pyridine, 8.33 g (0.073 mol) of vinyl butyrate, 9.52 g (0.073 mol) of cyclohexanecarboxylic acid, and 5.0 g of

an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40'C for 17.5 hours imder a slight flow of mtrogen and stirring was stopped. The enzyme was filtrated, and washed with 54.1 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile (8.6 g, 0.025 mol, yield: 34.4%, optical purity: 98.7% ee), cyclohexanecarboxylic acid salt was obtained, The combined toluene layer was washed five times with water (187.5 ml x 5) at 20°C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-{4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (7.6 g, 0.0221 mol, extraction yield: 89%), cyclohexanecarboxylic acid salt was obtained.
Example 15
(S)-4-[4-D™gthylamino-l- (4'fluorophenyl)-l -hydroxybutyll-3-hydroxymetfayl-benzonitrile o-toluic acid salt
A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 25.0 g (0.073 mol) of (±)-4-[4-dimethylamino-l-(4*-fluorophenyl)-l-hydroxybutyl]-3-hydroxyraethylbenzonitrile in toluene, and the temperature thereof was controlled so as to be 40^C. Then, 5.77 g (0.073 mol) of pyridine, 8.33 g (0.073 mol) of vinyl butyrate, 9.93 g (0.073 mol) of o-toluic acid, and 5.0 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40°C for 21 hours under a slight flow of nitrogen, and then stirring was stopped The enzyme was filtrated firom the reaction mixture with Kiriyama funnel, and washed with 54.1 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (9.7 g, 0.028 mol, yield: 38.8%, optical purity: 97.8% ee), o-toluic acid salt was obtained.
The combined toluene layer was washed three times with water (250 ml, 63 ml, 63 ml) at eO^'C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (7.8 g, 0.0228 mol, extraction yield: 80%), o-toluic acid salt was obtained.

Example 16
(S)-4-f4-Dimethylaniino-l-(4'-fluoropbenyl)-l-hydroxybutyI1-3-hydroxymethyl-benzonitrile isobutyric acid salt
A four-necked flask equipped with stirrer and thermometer was charged with 33.3 g of a solution of 10.0 g (0.029 mol) of (i)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile in toluene, and die temperature thereof was controlled so as to be 40T. Then, 2.29 g (0.029 mol) of pyridine, 2.57 g (0.029 mol) of iso-butyric acid, 3.33 g (0.029 mol) of vinyl butyrate, and 2.0 g of an immobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40**C for 24 hours under a slight flow of nitrogen and stirring was stopped. The enzyme was filtrated off firom the reaction mixture with Kiriyama funnel, and washed with 2L6 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylamino-1 -(4'-fluoropheny 1)-1 -hydroxybutyl] -. 3-hydroxymethylbenzonitrile (3.8 g, 0.011 mol, yield: 38.0%, optical purity: 95.9% ee), isobutyric acid salt was obtained.
Example 17
(S)-4-[4-Dimethylamino-l- (4'-fluorophenyl)-l -hydroxybutyl]-3-hydroxymethyl-benzonitrile benzoic acid salt
A four-necked flask equipped with stirrer and thermometer was charged with 250.0 g of a solution of 50.0 g (0.146 mol) of (i)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile in toluene, and the temperature tiiereof was controlled so as to be 40°C. Then, 11.5 g (0.146 mol) of pyridine, 33.2 g (0.291 mol) of vinyl butyrate, 17.8 g (0.146 mol) of benzoic acid, and 10.0 g of an irmnobilized enzyme (Novozym 435) was added into the above mixture. The reaction mixture was stirred at 40°C for 20 hours under a sUght flow of nitrogen, and then stirring was stopped. The enzyme was filtrated firom the reaction mixture with Kiriyama fiinnel, and washed with 108 g of toluene. As a result, a toluene layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyI]-

3-hydroxymethylbenzonitrile (22.0 g, 0.064 mol, yield: 44.0%, optical purity: 99.0% ee), benzoic acid salt was obtained.
The combined toluene layer was washed three times with water (500 ml x 3) at eO'^C, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4-fluorophenyl)-!-hydroxybutyl]"3-hydroxymethyibenzonitrile (21,0 g, 0.0614 mol, extraction yield: 96yo), benzoic acid salt was obtained.
The following examples 18 to 28 illustrate the separation of S-diol from the reaction mixture.
Example 18
A test tube equipped with a stopper was charged with 4.3 mg (0,0126 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (compound A), 6.8 mg (0.0165 mmol) of (R)-5-cyano-2-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-benzyl butyrate (compound B), 1 ml of toluene, 1 ml of water, and 0.291 m mol of various kinds of acids (in the case of a dibasic acid, 0.146 m mol is used), and stirred at 40°C for 1 hour. Thus-obtained mixed solvent was separated into aqueous layer and toluene layer, and then the concentrations of compound A and B in each layer were measured. Partition coefficients Ka or Kb are calculated according to the following equation. The results are shown in Table 11. Ka = (Concentration of the compound A in aqueous layer) / (Concentration of the compound A in toluene layer)
Kb = (Concentration of the compound B in aqueous layer) / (Concentration of the compound B in toluene layer)

Example 19
(S>4-r4-Diniethylagiino-l-(4'-fluorophenylVl-hydroxybmylV3-hydroxy^ benzonitrile pivalic acid salt
The enzyme was filtrated fi"om the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 1174.3 g of a toluene solution containing (S)- 4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (44.8 g, 0.131 mol), pivalic acid salt was added 600.3 g of water and extracted. Further twice extraction with 359.7 g of water gave 1375.9 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hyd^oxymethylben2;onitrile, pivahc acid salt. The aqueous solution contained 43.8 g (0.128 mol, yield: 97.8%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyI]-3-hydroxymethylbenzonitrile. Optical purity: 98.4%ee, (R)-5-cyano-2-[dimethyIamino-(4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 10,7 area%.
Example 20
(S)-4-[4-Dimethylamino-l - (4'-fluorophenyI)-1 -hydroxybutyl]-3-hydroxymethyl-benzonitrile pivalic acid salt
To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3"hydroxymethylbenzonitrile pivalic acid salt (content of (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzonitiile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.018 mol) of ammonium acetate. Then LO g of toluene was added therein and the mixture was stiired for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile (0.389 g, 1.136 mmol, yield: 97.3%), pivalic acid salt was obtained By HPLC analysis, it was found that chemical purity was 97.3 area % and (R)-5-cyano-2-[dimethylamino- (4'-fIuorophenyI)-hydroxybutyl] benzyl butyrate: 1.5 area % was contained.

Example 21
(SM-r4-Dimethylamino-l-(4'-fluorophenyIVl-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt
To 12.6 g of an aqueous solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethyIamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hydroxymethylbCT2X)nitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.013 mol) of lithium sulfate. Then 1,0 g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile, pivalic acid salt (0.394 g, 1.151 mmol, yield: 98.5%) was obtained. By HPLC analysis, it was found that chemical purity was 93.7 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 5.5 area % was contained.
Example 22
(S>-4-[4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt
To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile:
0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 1.4 g (0.011 mol) of ammonium sulfate. Then 1.0 g of toluene was added therein and the mixture was stirred for 10 miiiutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt (0.397 g, 1.159 mmol, yield: 99.3%) was obtained. By HPLC analysis, it was found that chemical purity was

94.7 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluoiophenyl)-hydroxybutyl] ben2yl butyrate: 4.5 area%, was contained
Example 23
(S)-4"r4-DimethylaminO'l-(4'-fIuorophenylVl-hydroxyb\ityl]-3-hydroxymethyl-Benzonitrile pivalic acid salt
To 12.6 g of an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethyIbenzonitrile pivalic acid salt {content of (S)-4-[4-dimethylamino-1 -(4'-£luorophenyI)-1 -hydroxybutyl]-3-hydroxymethylbenzonitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the mefliod of Example 19, was added with L4 g (0.0099 mol) of sodium sulfate. Then LO g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating the mixture solution, and an aqueous layer containing pivalic acid salt of (S)-4-[4-dimethylamino-l-(4*-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbeiizonitrile (0.391 g, 1.142 mmol, yield: 97.8%) was obtained By HPLC analysis, it was found that chemical purity was 94.5 area % and (R)-5-cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate: 4.7 area % was contained.
Example 24
(S)-4'C4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroKybutyl1-3-hydroxymethyl-benzonitrile pivalic acid salt
To 12.6 g of an aqueous solution of (S)-4-[4-dimethyiainino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzomtrile pivalic acid salt (content of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile: 0.4 g, 1.168 mmol}, which was separately obtained according to the method of Example 19, was added with 0.7 g (0.012 mol)of sodium chloride. Then 1.0 g of toluene was added therein and the mixture was stirred for 10 minutes. After standing for 30 minutes, the organic layer was discarded by separating

the mixture solution, and an aqueous layer containing pivalic acid salt of (S)-4-[4-dimethyIamino-1^4'-fluorophenyl>l-hydroxybutyl]-
3-hydroxymethylbenzomtriIe (0.397 g, 1.159 mmol, yield 99.3%) was obtained. By HPLC analysis, it was found that chemical purity was 93.3 area % and (R)-5-cyano-2-[dimethylamino- (4'-fluorophenyI)-hydroxybutyI] benzyl butyrate: 5.8 area % was contained.
Example 20-24 compared to example 19 show that the separation improves by the addition of salts.
Example 25
(S)-4-[4-Dimethylaniino-l-(4'fluorophenylVl-hydroxybutyll-3-hydroxymethyl-benzonitrile pivahc acid salt
The enzyme was filtrated firom the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.92 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid was adjusted so as to be 40 °C and 13 mL of water was added to the solution and extracted. The solution was further extracted twice with 7.5 mL of water, to obtain a solution of (S)-4-[4-dimethyIaniino-l-(4'-£Iuorophenyl)-l-hydroxybutyl]-3-hydroxymetfaylbenzonitrile, pivalic acid salt. In the above aqueous solution, 0.886 g (2.588 nmiol, yield: 88.6%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3'hydroxymethylbenzonitrile was contained.
Example 26
(S)-4-[4"Dimethylamino-l-(4^fluorophenyl)-l"hydroxybutyl1'3-hydroxymethyl-benzonitrile pivalic acid salt
The enzyme was filtrated firom the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.920 mmol) of (S)-4-[4-dimethylamino-l-(4'-

f
f[uorophenyl)-l-hydroxybutyl]-3-hydroxymethylbOT2om1iile, pivahc acid was adjusted so as to be 40°C and 11.6 mL of water was added to the solution and extracted. The solution was extracted further three times with 5.8 mL of water to obtain a solution of (S)-4-[4-dimethyIamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt. In the above aqueous solution, 0.879 g (2.567 mmol, yield 87.9%) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)4-hydroxybutyl]-3-hydroxymethyibenzonitrile was contained.
Example 27
(S)-4-[4-Dimethylamino-1 - (4' -fluorophenyi)-1 -hydroxybutyl]-3 -hydroxymethyl-benzonitrile pivalic acid salt
The enzyme was filtrated fi-om the enzyme reaction mixture obtaiaed by the method according to Example 1, and washed twice with toluene. Thus-obtained 10.4 g of toluene solution containing 1.0 g (2.92 mmol) of (S)-4-[4-dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivahc acid was added 13.0 mL of water and extracted. The solution was extracted further twice with 7.5 mL of water to obtain an aqueous solution of (S)■4-[4-dimethylamino-l-(4'-fluorophenyl)-I-hydroxybutyI]-3-ydroxymethylbenzonitriIe, pivalic acid salt. In the above aqueous solution, 0.954 g (2.786 mmol, yield 95.4%) of (S)-4-[4-dirQethylaniiQO-l-(4*-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile was contained.
Example 28
(S)-4-[4-Dimethylamino-l-(4^-fluorophenyl)-l-hydroxybutyl>3-hydroxymethyl-benzonitrile pivahc acid salt
The enzyme was filtrated from the enzyme reaction mixture obtained by the method according to Example 1, and washed twice with toluene. Thus-obtained 19.4 g of a toluene solution containing 1.0 g (2.90 mmol) of (S)-4-[4-dimethylamino-l-

(4'-fluorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivalic acid salt was added with 0.43 g (4.210 mmol) of pivalic acid and the mixture was extracted with 13.0 mL of water. The solution was fiirther extracted twice with 7.5 mL of water to obtain an aqueous solution of (S)-4-[4-dimethylamino-l-(4'-fIuorophenyl)-l-hydroxybutyl]-3-hydroxymethylbenzonitrile, pivaUc acid salt In the above aqueous solution, 0.947 g (2.766 mmol, yield: 94.7%) of (S)-4-[4-dimethylamino-l-(4'-fluo^ophenyl)-l-hydroxybutyI]-3-hydroxymethylbe^zonit^ile was contained.
A comparison of example 25, 26, 27 and 28 show that an improved separation is obtained by addition of pivalic acid whereas more washings and regulation of the temperature does not have great influence on the separation-Example 29
(S>4-[4'DimethylaminO" 1 -(4' -fluorophenyl)-! -hydroxybutyll-3-hydroxymethyl-benzonitrile
To a stirred solution of racemic 4-[4-DimethyIamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-ben2onitrile (0,29 mmol, 100 mg) and vinylbutyrate (3,9 mmol, 0,5 ml) in anhydrous 1,4-dioxane (2,5 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 5 mg). The reaction is heated to 37 °C and followed by HPLC. After 162 hours (at a conversion of 33,9%) the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 72% ee and (S)-diol with 28% ee.
Example 30
(S>4-[4-Dimethylamino-1 -(4' -fluorophenyl)-1 -hydroxybuty 1] -3 -hydroxymethy I-benzonitrile
To a stirred solution of racemic 4-[4-Dimethylamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzonitrile (0,29 mmol, 100 rag) and vinylbutyrate (0,58 nmiol, 73,6 |J.l) in-anhydrous 1,4-dioxane (3,0 ml) is added lipoprotein lipase

pseudomonas sp. (160 U, 4 mg). The reaction is heated to 37 °C and followed by HPLC. After 194 hours at a conversion of 18,7 %, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are ev^orated iji vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 92% ee and (S)-diol with 14% ee. The reaction was also monitored using 1, 4 and 8 equivalent vinylbutyrate as shown in table 12.

Example 31
(S)-4-[4-Dimethylaimno-l-(4'-fluorophenylVl-hydroxybutyl]-3-hydr^ benzonitrile
To a stirred solution of racemic 4-[4-Diinethylamino-l-(4-fluoro-pheiiyl)-l-hy(iroxy-butylj-S-hydroxymethyl-benzonitrUe (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 jil) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 5 mg). The reaction is heated to 50 ^^C and followed by HPLC. After 165 hours at a conversion of 30,4 %, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The .combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester with 98,1% ee and (S)-diol with 30,3% ee. The reaction was also monitored at 25, 37 and 65 °C as shown in table 13.

Example 32
(S>4-r4-Dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyl]-3-hy^^ benzonitrile
To a stiired solution of racemic 4-[4-Dimethylamino-l-(4-£Iuoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyI-benzonitrile (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 jil, 2, eq.) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 4,5 mg). The reaction is heated to 50 °C and followed by HPLC. After 209 hours, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography to give (R)-butyrate ester and (S)-did- The reaction was also monitored with (200 mg, 500 mg and 1000 mg diol and respectively 150 |il, 375 [il and 750 |il vinylbutyrate and 2,85 ml, 2,625 ml and 2,25 ml 1,4-dioxane as shown in table 14.

Example 33
(S)-4-[4-Dimethylamino-l-(4'-fluorophenyl)-l"hydroxybutyl]-3-hydroxy^ benzonitrile
To a stiired solution of racemic 4-[4-Diniethylamino--l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzonitrile (0,29 mmol, 100 mg) and vinylbutyrate (0,59 mmol, 75 |xl) in anhydrous 1,4-dioxane (2,925 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 4 mg). The reaction is heated to 50 °C and followed by HPLC. After 473 hours, the enzyme is filtered off and washed with a small amount of 1,4-dioxane. ^The combined organic phases are ev^orated in vacuo and subsequently analyzed HPLC. The result is shown in table 15.

Example 34
(SV4-['4-Dimethylamino-l-(4'-fluorophenyl)-l-hydroxybutyn-3-hydroxymethyl-benzonitrile

To^ stirred solution of racemic 4-[4-DimethyIamino-H4-fluoro--phenyl)-l-hydroxy"
V
butyI]-3-hyclroxymethyl-benzomtrile (2,9 mmol, 1000 mg) and vinylbutyrate (5,9 mmol, 750 fxl) in anhydrous 1,4-dioxane (14,25 ml) is added lipoprotein lipase pseudomonas sp. (160 U, 20 mg). The reaction is heated to 50 °C and followed by HPLC. After 139 hours, additional 5 mg lipase was added. After 155 hours, additional 15 mg lipase was added. After 399 hours, at a conversion of 55,7 % a sample was collected. The erLzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. After 560 hours at a conversion of 62,8 %, the reaction was stopped. The enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. The obtained ee-values are shown in table 16.

Example 35
(S)-citalopram diol analogues
To a stirred solution of racemic citalopram diol analogue (0,29 mmol, 100 mg) and vinylbutyrate (0,29 mmol, 37 |il) in anhydrous 1-4-dioxane (2,925 ml) is added 4-10 mg PspLL. -The reaction is heated to 50 degrees celcius and followed by HPLC. After -reaction was stopped, the enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 17.

Example 36
(S)-4-[4-Dimethylamino-1 -(4' -fluorophenyl)-1 -hydroxybutyl] -3-hydroxymethyl-benzonitrile
To a stiired solution of racemic 4-[4-Dimethylamiiio-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-beiizonitrile (29 mmol, 10 g) and vinylbutyrate (58 mmol, 7,5 ml) in anhydrous 1,4-dioxane (142,5 ml) is added lipoprotein lipase pseudomonas sp, (160 U, 250 mg). The reaction is heated to 50 °C and followed by HPLC. After 192 hours at a conversion of 41%, additional 250 mg lipase was added After 504 hours, at a conversion of 63 % the reaction was stopped. The enzyme is filtered off and washed with a small amount of 1,4-dioxane. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Obtained EE-value ((S-diol) = 95% (S-diol/R-diol = 40:1).
Example 37
Isolation of (S)-4-[4-Dimethylaniino-l-(4'-fluorophenyl)-l-hydroxybutyI1-3-
hydroxymethyl-benzonitrile by flash-cbromatography

A mixture of 372 mg S-citalopram diol with ee-value 94,5% and 628 mg R-citalopram diol butyrate ester with ee-value 59,5% was dissolved in a Tninimtim of ethylacetate/heptane 4:1 with 4% triethylamine. The S-citalopram diol was isolated using flash-chromatography in ethylacetate/heptane 4:1 with 4% triethylamine to obtained 120 mg S-citalopram diol (ee-value 94,5%).
Example 38
Isolation of (S)-4-r4-DimethylammO"l-(4'-fluorQphenyl)-l-hydroxybutyl1-3-hydroxymethyl-benzonitrile by washing a mixture containing the S-diol base
To a mixture of S-citalopram diol and R-citalopram diol butyrate ester (0,3g each) 'was added 24 ml heptane and 66 ml water/methanol (2:1). The clear solution was transferred to a separation fimnel and organic phase was collected. Additional 10 ml heptane was added to aqueous phase and mixed thoroughly. Organic phase was collected. Ekstraction repeated additional 4 times. The combined heptane phases were ekstracted with 20 ml water/methanol (2:1) and aqueous phases were combined and evaporated to half volume and extracted with 3 times 10 ml ethylacetate. The combined ethylacetate phases were dried with Na2S04, filtered and dried in vacuo to obtaine 0,15 g S-citalopram diol containing 2% R-citalopram diol butyrate ester.
Example 39
(S)-citalopram diol analogues using Novozymes 435
To a stirred solution of racemic citalopram diol analogue (0,29 mmol, 100 mg) and vinylbutyrate (0,29 mmol, 37 ^1) in anhydrous toluene (2,925 ml) is added 0,2 mg Novozymes 435 and (0,32 mmol, 33 mg) pivalic acid. The reaction is heated to 40 degrees celcius and followed by HPLC. After reaction was stopped, the enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 18.

Example 40
The influence of various carboxylic acids on the enzymatic acylation using Novozymes435
To a stirred solution of racemic 4-[4-Dimethylamino-l-(4-fluoro-phenyl)-l-hydroxy-butyl]-3-hydroxymethyl-benzomtrile (0,29 mmol, 100 mg) and vinylbutyrate (0,29 nmiol, 37 ^1) in anhydrous toluene (2,925 ml) is added Novozymes 435, (0,2 mg) and 1,1 eq. Carboxylic acid. The reaction is heated to 40 degrees celcius and followed by HPLC. The enzyme is filtered off and washed with a small amount of toluene. The combined organic phases are evaporated in vacuo and subsequently analyzed on super critical fluid chromatography. Result is shown in table 19.

Example 41
(R>-5-Cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyll hcnzyl butyrate
A four-necked flask equipped with stirrer and thermometer was charged with 219 g of a solution containing 21.9 g (0.064 mol) of (±)-4-[4-dimethylamino-l- (4*-fluorophenyl)-l-hydroxybutyl]-
3-hydroxymethylben2onitrile in toluene. Then, 14.6 g (0.128 mmol) of vinyl butyrate, 5.07 g (0.128 mol) of pyridine, 7.89 g (0.064 mol) of benzoic acid and 2.19 g of an immobilized enzyme (Novozym 435) were added into the above mixture. The reaction mixture was allowed to warm up to 60 °C and stirred for 15 hours under a slight flow of nitrogen and stirring was stopped. The enzyme was filtrated off from the reaction mixture with Kiriyama funnel, and washed with 50 g of toluene. The combined toluene layei: was washed twice with water (255 ml, 265 ml) and concentrated to give 14.9 g of (R)-5-Cyano-2- [dimethylamino- (4'-fluorophenyl)-hydroxybutyl] benzyl butyrate benzoic acid salt

To a mixture of 4,0 g of (R)-5"Cyano-2- [dimethylamino- (4'-fluoropheiiyl> flydroxybutyl] benzyl butyrate benzoic acid salt (7.48 mmol), 40 ml of water and 40 mi of toluene were added 1.26 g of 30% sodium hydroxide (9.45 mol) with stirring, the mixture was kept stirring for 30 minutes. After stirring, the mixture was allowed to stand and the toluene layer was separated. The toluene layer was washed with 40 ml of water and concentrated at 40 0C under reduced pressure, to thereby obtain 2.7 g of (R)-5-Cyano-2- [dimethylamino- (4-fluorophenyl)-hydroxybutyl] benzyl butyrate. IH NMR (400 MHz. CDC13) a(ppm): 7.69 (IH, s), 7.68-7.56 (2H, m), 7.36-7.23 (2H, m), 7.02-6.90 (2H, m), 5.41 (IH, d, J = 14.9 Hz), 4.99 (IH, d, J = 14.6 Hz), 2.59-2.42 (IH, m), 2.40-2.30 (3H, m),2.26 (2H, dt, J= 7.3 Hz, 1^ Hz), 2.18 (6H, s), 1.68-1.57 (2H, m), 1.67-1.46 (2H, m), 0.93 (3H, t, J = 7.6 Hz).
Example 42
Escitalopram,oxalate
(iS)-4-(4-Dimethylamino)-1-(4 '-fluorophenyl)-1 -hydroxybutyl)-3-hydroxymethylbenzonitrile (15.8 g, 46.2 mmol) having a chemical purity of 99% and an eeof98.7%asdetenninedby Chiral Supercritical Fluid (Chromatography, was dissolved in toluene (100 mL). Triethylamine (13.0 mL, 93.2 mmol) was added followed by slow addition of a solution of tosyl chloride (9.4 g, 49.4 lomol) in toluene (lOOmL). The resulting solution was stirred at room temperature for 20 min. then added water (50 mL) and cone. Ammonia (25 mL). The mixture was stiired at 40 0C for 2 rnin^ transferred to a separatory funnel. The phases were separated and the organic phase was washed with water (50 mL), dried with MgS04, filtered and concentrated under reduced pressure to give the cmde product (14.2 g) in 95% yield. The crude product was dissolved in ethanol (17 mL) and was added a solution of oxaUc acid (3.95 g, 43.9 mmol) in ethanol (27 mL). The precipitate was collected by filtration to give Escitalopram, oxalate (14.0 g), which was re-crystaUised from ethanol (85 mL) to give the final product (12.2 g). The purity was deteraiined to be 99.65% by HPLC. The ee was 98.5% deteraiined by Chiral Supercritical Fluid Chromatography.

WE CLAIM:
1. A method for the preparation of the S- or R-enantiomer of a diol having the formula

wherein R is cyano or a group which may he converted to a cyano group, Z is a group -CH2-N(R'R') wherein R'and R' are C1-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may he converted to a dimethylaminomethyl group, the dotted line is a double or single bond and Hal is halogen, or a salt thereof and/or the opposite
F
enantiomer of an acylated diol having the formula
wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R1, wherein R1 is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-10-allcyl)amino, aryl, aryloxy, arylthio and heteroaiyl, or R1 is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Cuio-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy,

s
C1-10-alkyithio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di"(C1-10- aIkyl)amino and Y is a bond, 0, S or NH, or a salt thereof comprising
a) subjecting a racemic compound of formula

wherein R, Z, the dotted line and Hal are as defined above, to selective enzymatic acylation using an acylating agent having the formula

or an isocyanate having the formula R1-N=C=0 or an isothiocyanate having the formula R1-N=C=S;
wherein X is O or S; W is O or S; U is O or S, V is halogen;
R° is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be
substituted one or more times with substituents selected from CMo-alkoxy, CMQ-
•
alkylthio, hydroxy, halogen, amino, nitro, cyano^ C1-10-alkylamino, di-(C1-10-alkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R° is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from Cuo-allcyl, C1-10-alkenyl, C1-10-allkny1 C1-10-alkoxy, CMQ-

alkjdthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(CMo-alkyI)amino;
R1 is as defined for R0;
R is as defined for R , or R is a suitable leaving group;
or R° and R^ together form a chain of 3 to 5 carbon atoms;
provided that W and U is not S when X is S; to fonn a mixture of tbe starting material of formula (II) in either the R- or the S-form and opposite enantiomer of a compound having the fonnula

wherein R, W, Hal, R3, the dotted line and Z are as defined above; or b) subjecting a racemic compound of formula

-1
wherein R, Z, W, Hal, the dotted line and R are as defined above; to selective enzymatic deacylation to form a mixture of deacylated compound of formula

wherein R, Hal, the dotted line and Z are as defined above in either the R- or the S-form and the acylated starting material of formula (IV) in the form of the opposite enantiomer;
optionally followed by, in either order, isolation of the S- or R-enantiomer of the compound of formula (II) and/or the opposite enantiomer of the compound of formula (IV) or a salt thereof

2. The method according to claim 1, wherein the acylation of the compound of formula (II) results in a mixture containing the compound of formula (II) in the S-form and the compound of formula (IV) in the R-form.
3. The method according to claim 1, wherein the acylation of the compound of formula (II) results in a mixture containing the compound of formula (II) in the R-form and the compound of formula (IV) in the S-form.
4. The method according to claim 1, wherein the deacylation of the
compound of formula (TV) results in a mixture containing the compound of formula (IV) in
the S-form and the compound of formula (II) in the R-form.
5. The method according to claim 1, wherein the deacylation of the compound of formula (IV) results in a mixture containing the compound of formula (IV) in the R-form and the compound of fonnula (II) in the S-form.
6. The method according to any of claims 1-5, wherein Hal is fluoro and R is halogen or cyano preferred R is cyano.
7. The method according to any of claims 1-6, wherein the dotted line represents a single bond.
8. The method according to any of claims 1-7 wherein and Z is
dimethylaminomethyl or a group that may be converted to a dimethylaminomethyl group,
preferred Z is a dimethylaminomethyl group.
9. The method according to any of claims 1, 2, 3 and 6-8, wherein the acylating agent is a compound selected from the compounds of formula (Ilia), (lUb) and (IIIc).
10. The method according to claim 9, wherein the acylating agent is a compound selected from the compounds of formula (Ilia) and (Illb).

n. ' The method according to claim 10 wherein the acylating agent is a compound of foraiula(ina).
IZ- The method according to claim IQ wherein the acylating agent is a compomid offoraiula(IIIb).
13. Tlie method accordmg to claim ^ wherein the acylating agent is a compomid of fonnula(inc).
14 The method according to any of claims 9 -11 wherein U is O.
15, The method according to any of claims 9-14 wherein W is O.
16. The method according to claims 9-15 wherein X is O.
n. The method according to any of claims 9-16 wherein R0, R1 and R2 are independently selected from CM-alkyl, C1-6-alkenyl and C2-4-alkynyl all of which may optionally be substituted one or more times with substituents selected from CM-alkoxy, CM-alkylthio, hydroxy, halogen, amino, nitro, cyano, CM-alkylamino and di-(CM-alkyl)amino or R2 is a leaving group, such as succinimidyl, HOBt and ipfp or R^ and R^ together form a chain of 3-5 carbon atoms.
\ 2. The method accordii^ to claim 17 wherein R0, R1 and R2 are independently selected from C1-3-alkyl, C2.3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di-(C1-3-alkyl)amino,
l9. The method according to claim Ig wherein R0 and R1 are C1-3-alkyl and R2 is C1-3-alkyl substituted one or more times with halogen or R2 is C2-3-alkenyL
%p. The method according to claim H wherein R0 and R1 are independently selected from C1-4-alkyl, C2-4-alkenyl and C2-4-alkynyl all of which may optionally be

substituted one or more times with substituents selected fiom C1-4-3Jkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(C1-4-alkyl)anxino.
Zl, The method according to claim 2D wherein R0 and R1 are
independently selected from Ct-s-alkyl, C2.3-alkenyl or C2-3-aIkynyI all of which may
all optionally be substituted one or more times with substituents selected from C1-3-
alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino and di-
(C1-3'aIkyl)amiiio.
ISL. The method according to claim 21 wherein R° and R^ are Ci-i-alkyl.
23. The method of claim 2J,.wherein R*^ and R^ are C1-3-aDcyl, smtable methyl, ethyl or propyl, preferred propyl.
24^ ♦ The method of claim 12- wherein R^ and R^ are independently selected from Ci-4-alkyl, C2-4-alkenyl and C2^-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-4-alkoxy, C1-6-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-allcylamino and di"(C1-6-alkyl)amino or R2 is a leaving group, such as succinimidyU HOBt and pfp.
25: The method of claim 24 wherem R1 is selected from C1-3-alkyl, C2.3-alkenyl or Cao-alkynyl all of which may optionally be substituted one or more times with substituents selected from C1-3-alkoxy, C1-3-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cu3-allcylamino and di-(C1-3-alkyl)amino and R2 is Cia-alkyl substituted or more times with halogen or R2 is C2-4-alkenyl.
26 The method according to claim 25"wherein R' is C1-3-alkyl, C1-3-alkenyl and C2-3-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci*.3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, C1-3-aIkylaniino and di-(C1-3-alkyl)amino and R2 is Cu3-allcyl substituted one or more times with halogen or R2 is Cao-alkenyl.
2-T The method according to claim 25'wherein R1 is d-s-alkyl.

23- The method according to claim 27 wherein R^ is C1-3-alkyl, such as methyl, ethyl or propyl and C],3-alkyl substituted one or more times with halogen or R is €2-3-alkenyl,
11. The method according to claim Z8 wherein R^ is vinyl.
3(X The method according to claim 251 wherein R^ is propyl.
31. The method according to claim 13 wherein R^ is C1-6-alkyl, C2-*-alkenyl and C2^-alkynyl all of which may optionally be substituted one or more times with substituents selected from Ci-4-aIkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci-4-alkylamino and di-(C1-4-aIkyl)amino,
HL. The method according to claim 3t wherein R^ is Cua-alkyU Ci-j-alkenyl and C2-3-aJkynyl all of which may optionally be substituted one or more times with C1.3-alkoxy, C1-3-alkylthio. hydroxy, halogen, amino, nitro, cyano, C1-3-alkylamino^and di-(C1-3-2ilkyl)amino.
33. The method according to claim 3Zwherein R^ is Ci,3-alkyl, C2.3"alkenyl or C2-3-alkynyl preferred R^ is C1-3-alkyl, such as methyl, ethyl or propyl, prefeired propyl
3ih The method according to any of claims 1, 2-3 and 6-^5 wherein the acylating agent is an isocyanate of foraiula R^-N= 35". The method according to claim 3/^ wherein the acylating agent is an isothiocyanate of the formula R^-N=C=S.
36- The method according to 'claim Jk- wherein the acylating reagent is an isocyanate of formula R^-N==C=0.
37 The method according to any of claims 3/^-36 wherein R^ is C1-4-alkyl, C1-6-alkenyl or Cj^-alkynyl all of wfech may optionally be substituted one or more times

with substituents selected from C1-4-alkoxy, C1-4-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-4-alkylamino and di-(C1-4-alkyl)aniino.
3?- The method according to any of claims 37 wherein R^ is C1-3-alkyl, C2-3-alkenyl or C2-3-alkynyl all of which may optionally be substituted one or more times with C1-3-alkoxy, C1-3-aIkylthio, hydroxy, halogen, amino, nitro, cyano, C1.3-alkylamino and di-(C1-3-alkyl)aniino.
3^- The method according to claim?»8 wherein R^ is C1-6-alkyl, C2.3-aIkenyl or C2-3-alkynyL
40. The method of claim 3^ wherein R^ is methyl, ethyl, or propyl, preferredpropyl
41. The method according to any of claims 1 j and fy^Z wherein the racemic compound of formula (TV) used is a compound wherein Y is O, or S.
44v The method according to claim 41 wherein the racemic compoimd of formula (TV) is a compound wherein Y is 0.
4S. The method according to claim 41 wherein the racemic compoimd of formula (IV) used is a compound wherein Y is S.
4if- The method according to any of claims 1 ^ and tf- % wherein the racemic compound of foraiula (IV) used is a compound wherein Y is a bond.
45. The method according to any of claims 1 . and i^^t wherein the racemic compound of foraiula (IV) used is a compound wherein Y is NH.
46. The method according to any of claims 4J-4S" wherein R is Ci-10-alkyl, C2-10-alkenyl or C2-10-alkynyI all of which may optionally be substituted one or more times with substituents selected from C1-10-alkoxy, C1-10-aUcylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino.

47» The method of claim 46 wherein R^ is C1-10-alkyl, C1-10-alkenyl or C2-10-aDcynyl all of which may optionally be substituted one or more times with substituents selected from hydroxy, halogen, amino, nitro and cyano.
/f 2. The method according to claim 47 wherein R^ is C1-10-alkyL
l^^^ The method according to any of claims l^^nS and g-J^o wherein the enzymatic acylation is performed with a hydrolase, such as a hpase, an esterase, an acylase or a protease,
50. The method according to any of claims l.j^-.^.* and 3 ?'4g wherein the
enzymatic deacylation is performed with a hydrolase, such as a Upase, an esterase, an
acylase or a protease.
51, The method according to any of claims tfff^SO wherein the hydrolase is used in
the forai of an immobilized enzyme or Cross-Linked Enzyme Crystal (CLEC)
enzymes.
5%, The method according to any of claims Lp^-St wherein the enzyme used is selected from Pseudomonas sp, hpoprotein Upase, Candida antartica Upase B and Thermomyces lanuginosus Upase or a mutant or variant of these enzymes.
53. The method according to any of claims ^9--^wherein the enzyme is Pseudomonas sp. Upoprotein Upase or a mutant or variant of Pseudomonas sp. Upoprotein Upase.
5 If, The method according to claim 5S wherein the enzyme used is Pseudomonas sp. Upoprotein Upase.
557 The niethod according to any of claims/jf^-ica. wherein the enzyme is Candida antartica lipase B or a mutant or variant of Candida antartica lipase B.
56, The method according to claim Si'wherein the enzyme is Candida antartica Upase B.

57. The method according to claim 56 wherein the enzyme is Novo2yme®435.
S^, The method according to any of claims i^^-St wherein the enzyme is Theimomyces lanuginosus lipase or a mutant or variant of Theraiomyces lanuginosus lipase.
S^^ The method according to claim 5^ wherein the enzyme is Themiomyces lanuginosus hpase.
) 60* The method according to claim SJ wherein the enzyme is Lypozymc^^ TL
IM.
6J, The method according to any of claims 1-66 wherein the enzymatic acylation or the enzymatic deacylation is carried out in presence of an organic base or an organic acid or a mixture thereof.
62^ The method according to claim Si A^herein the enzymatic acylation or enzymatic deacylation is carried out in the presence of an organic acid.
63. The method according to claim ^wherein the enzymatic acylation is carried out in presence of an organic acid.
6i^, The method according to any of claims 62^3 wherein the organic acid is an aromatic carboxylic acid or an aUphatic caiboxyUc acid.
6S; The method according to any of claims 61-64 "^^^^^ ^® organic acid is selected from n-propionic acid, iso-propionic acid, n-butyric acid, iso-butyric acid, isO'Valeric acid, 2-ethylbutyric acid, cyclohexanecarboxylic acid, pivalic acid, benzoic acid, p-toluic acid, saUcylic acid and 3-phenylpropionic acid.
6^» The method according to claim 6& wherein the organic acid is pivalic acid.

67 A method for the isolation and purification of the compound having the formula

wherein R is cyano or a group which may be converted to a cyano group, the dotted hne represents a double or single bond, Hal is halogen, Z is a group -CH2-N(R'R') wherein R'and R' are C1-6-alkyl, or R'and R' are connected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a groiq) which may be converted to a dimethylaminomethyl group, W is O or S and R"^ is -Y-R^ wherein Y is a bond, O, S or NH and R^ is C1-10-alkyl C1-10-alkenyl or C2-1Q-alkynyl all of which may optionally be substituted with one or more substituents selected from C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, Ci. lo-alkylamino, di and/or the diol of formula

wherein R, Z, Hal and the dotted line are as defined above, or a salt thereof^ from a mixture containing the compound of formula (IV) and the diol of formula (II), which comprises:
a) treating said mixture contaimng the compound of formula
(IV) and the diol of formula (II) in a mixture of water and an organic solvent in the presence of an acid;
b) separating the aqueous phase containing the diol of fonnula (H) as a salt of
said acid from the organic phase to obtain an organic phase containing the compound
of fonnula (IV) as a salt of said acid; and
optionally isolating the compound of formula (II) as the base or as a salt thereof and optionally isolation of the compound of formula (IV) as the base or a salt thereof.
6^, The method according to claim f>1 wherein the S-enantiomer of the diol of formula (H) is separated from the R-enantiomer of the acyl derivative of formula
(IV).
Cn- The method according to claim gy wherein the S-enantiomer of the acyl derivative of fonnula (IV) is separated from the R-enantiomer of the diol of formula
(n).
10« The method according to claims ft7-0 wherein the mixture of the R- or S-enantiomer of an acyl derivative of fonnula (IV) and the opposite enantiomer of an diol of fonnula (II) used in step a) has been prepared according to the method in claims 1-62

71. A method for isolation and purification of flie acyl derivative having the fonnula

wherein R is cyano or a group which may be converted to a cyano group, Hal is halogen, the dotted line represents a double or a single bond, Z is a group -CH2-N(RTI'0 wherein R'and R" are Ci-6-aIkyl, or R'and R" araconnected to each other to form a cyclic structure including the N-atom to which they are attached, or Z is a group which may be converted to a dimethylaminomethyl groitp, W is O or S; and R^ is -Y-R^ wherin Y is a bond, 0, S or NH and R^ is C1-4o-alkyl, C2-10-aUcenyl or C2-lo-alkynyl all of which may optionally be substituted one or more times with substituents selected from Cj.io-alkoxy, Ci-10-allsyltiuo, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-10-aIkyI)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-4O-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-4o-alkylthio, hydroxy, halogen, amino, nitro, cyano, Cj.jo-aUcylamino and di-(C1-4o-alkyl)amino, or a salt thereof
and/or the diol of formula

wherein R, Hal, Z and the dotted line are as defined above, from a mixture containing the acyl derivative of fonnula (TV) and the diol of formula (II), which comprises:
a) treating said mixture containing the acyl derivative of formula (TV) and the diol of fonnula (U) in a mixture of water, a protic organic solvent and an apolar organic solvent;
b) separating the aqueous phase containing the diol of formula (II), from the organic phase to obtain an organic phase containing the acyl derivative of formula (TV); and
optionally isolating the diol of fonnula (IT) and/or the compound of formula (IV) from the aqueous/organic ohase, and optionally conversion of the compound of foraiula (H) and /or (TV) to salt thereof.
72. The method according to claim 71 wherein the S-diol of the diol of fonnula (II) is separated from the R-enantiomer of the acyl derivative of formula (IV).
73. The method according to claim 7j wherein the S-enantiomer of the acyl
derivative of fonnula (IV) is separated from the R-enantiomer of the diol of fonnula
(H).
74. The method according to claims IhTS wherein the mixtuxe of the R- or S-
enantiomer of a compound of formula (IV) and the opposite enantiomer of a
compound of formula (I[) used in step a) has been prepared according to the method
in claims 1-62

7S- A process for the preparation of escitalopram having the formula

or a phannaceutically acceptable salt thereof comprising preparation of the S-enantiomer of a diol having the formula

wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond, Z is a dimethylaminomethyl group or a group which may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof,
or the S-enantiomer of an acylated diol having the formula

wherein R, Z, the dotted line and Hal are as defined above, W is O or S, and R^ is -Y-R\ wherein R^ is C1-10-alkyl, C2-10-alkenyl or C2-10-alkyiiyl all of which may optionally be substituted one or more times with substituents selected firom C1-4O-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(Ci-i(ralkyl)amino, aryl, aryloxy, arylthio and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected firom C1-10-alkyl, C1-10-alkenyl, C2-i(ralkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, O, S or NH, or a salt thereof, by the method in any of claims 1-62, optionally followed by, in either order, conversion of the group R to a cyano grqup, conversion of the group Z to a dimethylaminomethyl group, reduction of a double boiid represented by the dotted line to a single bond and /or conversion of the group Hal to a fluoro group and then ringclosure under basic conditions of the S-enantiomer of foraiula (lis) or (TVs) or a labile ester derivative thereof to form a compound of foraiula

^
followed by in either order, if R is not cyano conversion of the group R to a cyano groiq), if Z is not dimethylaniinomethyl conversion of the group Z to a dimethylaminomethyl group, if the dotted line represents a double bond reduction to a single bond and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolation of escitalopram or a pharmaceutically acceptable salt thereof.
7^ The method according to claim 7S wherein the rrdxture of the R- or S-enantiomer of a compound of formula (H) and the opposite enantiomcr of the compound of formula (IV) obtained by emymatic acylation has been separated from each other by the isolation and purification process according to any of claims 84-87.
77 - The method according to claim is" wherein the R- or S-enantiomer of a compound cf formula (H) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 88-91,
-^^ A method for the preparation of racemic citalopram and /or escitalopram or a pharmaceutically acceptable salt thereof comprising preparation of the R-enantiomer of a diol having the formula

wherein R is cyano or a group which may be converted to a cyano group, the dotted line represents a double or a single bond and Z is a dimethylaminomethyl group or a group wl-vich may be converted to a dimethylaminomethyl group and Hal is halogen, or a salt thereof.

or the R-aiantiomer of an acylated diol having the formula

wherein R, Z, the dotted line Hal are as defined above, W is O or S, and R^ is -Y-R\ wherein R^ is C1-10-alkyl, C2-10-alkenyl or C2-10-alkynyl all of which may optionally be substituted one or more times with substituents selected firom C1-10-alkoxy, C1-4O-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino, di-(C1-4o-alkyl)amino, aryl, aryloxy, arylthid and heteroaryl, or R^ is aryl, wherein any of the aryl and heteroaryl groups may optionally be substituted one or more times with substituents selected from C1-10-alkyl, C2-10-alkenyl, C2-10-alkynyl, C1-10-alkoxy, C1-10-alkylthio, hydroxy, halogen, amino, nitro, cyano, C1-10-alkylamino and di-(C1-10-alkyl)amino and Y is a bond, 0, S or NH, or a salt thereof, by the method according to any of claims 1-62, optionally followed by, in either order, conversion of the group R to a cyano group, reduction of a double bond represented by the dotted line to a single bond, conversion of the group Z to a dimethylaminomethyl group and /or conversion of the group Hal to a fluoro group and then ringclosure under acidic conditions of the R-enantiomer of formula (Ilr) or (TVs) to form a mixture of the S-enantiomer of the compound of formula

and a minor amount of the corresponding R-enantiomer, followed by, in either order, if R is not cyano conversion of the group R to a cyano group and, if Z is not dimethylaminomethyl conversion of the group Z to a dimethylaminomethyl group, if the dotted line represents a double bond reduction to form a single bond and if Hal is not fluoro, conversion of Hal to a fluoro group, followed by isolatiC1-4 of escitalopram and/ or racemic citalopram or a phaimaceuticaliy acceptable salt thereof.
79. The method according to claim 78 wherein the racemic citalopram is isolated by precipitation of racemic citalopram free base or a salt thereof and escitalopram is recovered from the mother liquor of the precipitation.
80* The method according to claim "7^ wherein the mixture of the R- or S-enantiomer of a compound of formula (D) and the opposite enantiomer of the compoimd of formula (IV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 84-87.
8) The method according to claim 7^ wherein the R- or S-enantiomcr of a compound of formula (II) and the opposite enantiomer of the compound of formula (TV) obtained by enzymatic acylation has been separated from each other by the isolation and purification process according to any of claims 88-91.

Documents:

0367-chenp-2005 abstract duplicate.pdf

0367-chenp-2005 claims duplicate.pdf

0367-chenp-2005 description (complete) duplicate.pdf

367-chenp-2005-abstract.pdf

367-chenp-2005-claims.pdf

367-chenp-2005-correspondnece-others.pdf

367-chenp-2005-correspondnece-po.pdf

367-chenp-2005-description(complete).pdf

367-chenp-2005-form 1.pdf

367-chenp-2005-form 18.pdf

367-chenp-2005-form 26.pdf

367-chenp-2005-form 3.pdf

367-chenp-2005-form 5.pdf

367-chenp-2005-pct.pdf

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Patent Number

221245

Indian Patent Application Number

367/CHENP/2005

PG Journal Number

31/2008

Publication Date

01-Aug-2008

Grant Date

20-Jun-2008

Date of Filing

10-Mar-2005

Name of Patentee

H. LUNDBECK A/S

Applicant Address

9, OTTILIAVEJ, DK-2500 VALBY-COPENHAGEN,

Inventors:

#	Inventor's Name	Inventor's Address
1	TAOKA, NAOAKI	1-1-1210 DOUSYOUDAI, 1-CHOME, SUMA-KU KOBE, HYOGO, 654-0122,
2	YAMAMOTO, SHOGO	SANSEISO, 31-17 SHIYACHO 6-CHOME, TARUMI-KU KOBE, HYOGO 655-0872,
3	TAKEDA, Toshihiro	11-7, Ihozakiminami, Takasago, Hyogo 676-0078,
4	DANCER, Robert	Sønderkær 53, DK-2650 Hvidovre,
5	LYNGSO, Lars Ole	Aldershvilevej 121A, st.th., DK-2880 Bagsværd,
6	YOSHIDA, Takashi	Sanseiso, 31-17, Shioyacho, 6-chome, Tarumi-ku, Kobe, Hyogo 655-0872,
7	KATO, TAKAHISA	51-2-205 KOKUBO, AKASHI, HYOGO 673-0005,
8	UEDA, Yasuyoshi	140-15, Waku, Aboshi-ku, Himeji, Hyogo 671-1227,
9	PETERSEN, Hans	Guldagervej 11, DK-2720 Vanlose,
10	AHMADIAN, Haleh	Digeparken 12, DK-2680 Solrød Strand,

PCT International Classification Number

C07B 57/00

PCT International Application Number

PCT/DK03/00537

PCT International Filing date

2003-08-12

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	PA 2002 01201	2002-08-12	Denmark