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DESCRIPTION CYCLQALKYL-SUBSTITUTED AMINOMETHYLPRBROLIDINE DERIVATIVE
TECHNICAL FIELD
This invention relates to a synthetic quinolone antibacterial agent useful as a drug for humans, animals or fishes or an antibacterial preservative-
This invention also relates to a synthetic cpiinolone antibacterial agent in which the structure of substituent at the 7-position of l,4-dihydro-4-oxoquinoline skeleton or at the 10-position of 2,3"dihydro-7*oxo-7H-pyrido[l,2,3-de] [1.4]benzoxazine skeleton exerts inportant influence xxpan the expression of pharmacological effects such as ari-hTVurH-^-iHai activity, pharmacokinetics and safety, having a 3-[1-amino-l-cycloalkyl]methylpyrrolidin-l-yl qroup which can provide excellent antibacterial activity, pharmacokinetics and safety, as a sxabstituent at the 7- or 10-position, and also having excellent antibacterial activity, proper pharmacokinetics and high safety, namely to a 6-fluoro-l-[2-(S)-fluoro-l-(R)-cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carbo3^1ic acid derivative or a 2,3-dihydro-3- (S) -methyl"7-oxo-7H-pyrido [1,2,3-de] [1,4]benzoxazine-6-carboxylic acid derivative, and to an antibacterial agent and an antibacterial preparation, which con^tain the con5>ound.
BACKGROUND ART
Since the discovery of Norfloxacin, antibacterial activity and pharmacokinetics of synthetic qainolone antibacterial agents have been isproved, and many ccopoiinds are now used in the clinical field as chemotherapeutic agents which are effective in almost systemic infectious diseases.
In recent years, generation of bacteria having low sensitivity to synthetic quinolone antibacterial agents has been increasing in the field of clinics. For exarnple, like the case of Staphylococcus aureus (ME^SA) and pneumococcus (FR5P) idiich are non-sensitive to p-lactam antibiotics and enterococcus (VBE) which is non-sensitive to aminoglycoside antibacterial agents, a case has been increasing in which a Gram-positive bacterium originally resistant to drugs other than synthetic quinolone antibacterial agents becomes low-sensitive to synthetic quinolone antibacterial agents too. In consequence, development of a drug having further high efficacy has been called for in the field of clinics. On the other hand, it has been revealed that synthetic quinolone antibacterial agents cause a side effect in which convulsion is induced when a non-steroidal anti-inflammatory drug is simultaneously used, as well as other side effects such as phototoxicity, so that development of a synthetic quinolone antibacterial agent having further high safety has also been called for in the field.
It is known that structures of substituents at the 7-position and 1-position have a great influence to the antibacterial activity, pharmacokinetics and safety of synthetic
quinolone antibacterial agents. It is already known that guinolone derivatives having 3-ami ncanethylpyrrolidine as a STabstituent show strong antibacterial activity for Gram-negative and Gram-positive bacteria. For example, a 7"(3-ami nomethylpyrrolidin-1-yl) cjainolonecarboxylic acid derivative is described in Journal of Medicinal Cbesaistxy, vol. 29, p. 445 (1986), a 7-[3-(l-amino-l-methylethyl)pyrrolidin-l-yl) quinolonecarbo3cylic acid derivative is described in JovumaZ of Medicinal Chemistry, vol. 37, p. 733 (1994), and a 7-[3-(l-aminoal3cyl)pyrrolidi n-l-yl] cpainolonecarboacylic acid derivative is described in riitfgm'/^ja? £ Pharmacenit.ifvi7 Bulletin, vol. 42, p. 1442 (1994) . However, no compounds are known which have a 3-(l-amino-l-cycloalkyl)inethylpyridin-l-yl group at the 7-position and are also related to the present invention.
On the other hand, quinolone derivatives having 3-aminosnethylpyrrolidine as a substituent are compounds which show strong antibacterial activity, but, since most of these confounds have low selective toxicity, they act upon not only bacteria but also eucaryotic cells so that it is difficult to use them as medical drugs or animal drugs.
Also, it is known that quinolone derivatives having a 3-aminops'xrolidine derivative at the 7-position and 2- (S) -f luoro-1- (R) -cyclopropyl groi^ at the 1-position of the quinoline skeleton have weaker micronucleus inducing toxicity than those corresponding 1-cyclopropylquinolone derivatives. Their exanples are described in Journal of Medicinal Chemistry, vol.
37. P, 3344 (1994).
On the other hand, quinolonecarhojcylic acid derivatives having a 3-[l-amino-l"Cycloal]cyl]inethylpyrrolidin-l-yl grox:tp as a substituent, which are related to the present invention, are exen5>lified for example in JP-W-3-502452 (the term "JP-W" as used herein means an ^^unexaznined published Japanese international patent application") , and it describes compounds represented by a formula (a) or (b) shown below. However, substituent at the 5-position of these exeartplified quinolones is limited to a straight, branched or cyclic lower alkyl having 1 to 3 carbon atoms, and JP-W-3-502452 does not describe compounds having the l-[2-(S)-fluoro-l-(R)-cycloprppyl]quinoline skeleton or 3- (S) -methyl-7H-pyrido[1,2,3-de] [1.4]benzoxa2ine skeleton related to the present invention. In addition, JP-W-3-502452 does not disclose illustrative examples of the 3-[1-amino-l-cycloalJc^l ] methylpyrrolidin-1 -yl group. Formula (a)
[In the above formula, R7 is an alkyl having 1 to 4 carbon atoms, a vinyl, a haloalkyl, a hydroxyalkyl having 2 to 4 carbon atcms, a cycloaikyl having 3 to 6 carbon atoms, a phenyl or a phenyl substituted with a halogen, an alkyl, NH2 or OH, Re is a
straight, branched or cyclic lower alkyl having 1 to 3 carbon atoms, and X3 is CH, CF, CCl, CBr, N, CCF3, CSE2f CNO2, CR or COR^ (in these fozsiulae, R is a lower alkyl and R' is hydrogen or a lower all^l) . DefdLnitions of substituents of the cooqpound of formula (a) are independent to those the compound of the present invention.]
In the above formula, Z is a grot^ represented by the following formula (b) .
Formula (b)
(In this formula, m is an integer of from 0 to 4, and the substituents R9 and Rio are each independently a hydrogen atom, a lower alkyl or a cycloalkyl, Definitions of substituents of the compound of formula (b) are independent to those the coopound of the present invention.)
In addition, PCT WD 96/39407 discloses compounds represented by the following formula (c) , but they are limited to 2-pyridone derivatives such as 4H-4-oxoquinotozone skeleton, and PCT WO 96/39407 does not describe conpounds having the 1,4-dihydro-4-oxoquinoline skeleton or 2,3-dihydro"3-(S)-ethyl-7-oxo"7H-pyrido [1,2,3-de] [1.4] benzoxazine skeleton related to the
present invention. Also, PCT WO 96/39407 does not disclose illustrative examples of optically active 3-[l"aniino-l-cyclqprppyl]inethylpyrrolidin-l-yl group.
In addition, PCT WO 96/39407 does not describe about safety of the compounds of fozmula (c) .
Formula (c)
DISCLOSURE OF INVENTION
In view of the above, the inventors of the present invention have conducted intensive studies with the aim of providing the field of clinics with a con^jound which has excellent antibacterial activity, high efficacy and excellent safety. As a result of the extensive investigation, it has been found absolutely uneaq^ectedly that a cycloalkyl-substituted amincanethylpyrrolidine derivative represented by the formula (I) described below, its salts and hydrates thereof-can show strong antibacterial activity upon broad range of Gram-negative and Gram-positive bacteria, can show particularly strong anti bacterial activity aipon resistant strains of Gram-positive bacteria including ME^SA, PRSP and VPE, and also have excellent safety and good pharmacokinetics, thereby resulting in the
acconylishment of the present invention.
Particularly, it has been found that a cooqpound represented by the following formula (I) in which a cycloalkyl-substituted amincmethylpyrrolidine derivative is introduced at the 7-position of the l-[2-(S)-fluoro-l-(R)-
cyclopropyllquinoline skeleton, its salts and hydrates thereof show broad and excellent antibacterial activity iipon any one of Gram-negative and Gram-positive bacteria including drug-resistant strains, have excellent safety with sharply attenuated micronucleus induction action, and also have excellent pharmacokinetics.
Accordingly, the present invention relates to a compound represented by the following formula (I) , its salts and hydrates thereof:
{wherein R^ and R^ each independently represents a hydrogen atom or aa alkyl group having 1 to 6 carbon atoms, in which the alkyl group may have one or more substituents selected from the group consisting of a hydroxyl group, a halogen atom, an alkylthio group having 1 to 6 carbon atoms and an aUcyloxy group; n is an integer of 1 to 4; and
Q is a par-tial structure represented by the following forxDula (la):
[wherein R^ represents an all^l group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms/ a halogenoaUcyl group having 1 to 6 carbon atoms, a cyclic allcyl group having 3 to 6 carbon atoms which may have a substituent, an aryl group which may have a substituent, a heteroaryl group which may have a soabstituent, an alkosqfl group having 1 to 6 carbon atoms or an alkylamino group having 1 to 6 carbon atoms;
R"* represents a hydrogen atom or an alkylthio group having 1 to 6 carbon atoms;
R'* and the aforementioned R^ may form together with a part of the mother skeleton a ring structure optionally containing a sulfur atom as a ring constituting atom thereof and optionally having an alkyl groi:^) having 1 to 6 carbon atoms as a substituent;
R^ represents a hydrogen atom, an amino group, a hydroxyl group, a thiol groiip, a halogenomethyl grovqp, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an al3cynyl grovqp having 2 to 6 carbon atoms or an
alkoxyl grovip having 1 to 6 carbon atoms, in which the amino group may have one or more siibstituents selected from the group consisting of formyl group, an aJJcyl group having 1 to 6 carbon atoms and an acyl group having 2 to 5 carbon atoms;
X^ represents a halogen atom or a hydrogen atom, A"^ represents a nitrogen atom or a partial structure represented by formula (II) :
(wherein x" represents a hydrogen atom, an amuio group, a halogen atom, a cyano group, a halogenomethyl group, a halogenomethoii^l groi:^, an allcyl groiip having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms or an alkoaq^l grov^j having 1 to 6 carbon atoms, in which the amino group may have one or more st±>stituents selected from the group consisting of a fomyl group, an allcyl group having 1 to 6 carbon atoms and an acyl groxip having 2 to 5 carbon atoms; and
X^ and the aforementioned R"^ may form together with a part of the mother skeleton a ring structure optionally containing an oxygen atom, a nitrogen atom or a sulfur atom as a ring constituting atom thereof and optionally having an alkyl group having 1 to 6 carbon atoms as a substituent) ; and y represents a hydrogen atom, a phenyl grovp, an acetoxymethyl groi^, a pivaloyloxymethyl grot^j, an
etho3cynarhony 1 group, a choline group, a dimethylaiainoetfaYl group, a 5-indanyl groiap, a phthalidinyl group, a 5-aJJqfl-2-oxo-1,3-dioxol"4-Yl methyl gxovcp, a 3-acet03^-2-oxdbutyl groi:^, an alkyl group having 1 to 6 carbon atoms, an alkoacymethyl group having 2 to 7 carbon atoms or a phenylallc^l group conposed of an alkylene grooip having 1 to 6 carbon atoms and a i>henyl grot^] }.
The present invention also relates to each of the following items.
A cocpotmd, its salts and hydrates thereof, herein Q in the formula (I) is a 6-carboxy-9-fluoro-2,3-dihydro-3-(S)-methyl-7-oxo-7H-pyrido[l,2,3-de] I1.4]benzoxazin-10-yl group; the aforementioned compound, its salts and hydrates thereof, wherein the compound of formula (I) is a stereochemically pure coa;K>tind;
the aforementioned ccn^>ound, its salts and hydrates thereof, wherein R^ in the formula (I) is a halogenocyclqprqpyl group; the aforementioned compound, its salts and hydrates thereof, wherein the halogenocyclopropyl grotip din the formula (I) is a 1,2-cis-hcJ-ogenocyclpprqpyl group;
the aforementioned con^^ooond, its salts and hydrates thereof, wherein the halogenocyclopropyl group in the formula (X) is a stereochemically pure substituent:;
the aforementioned coopound, its salts and hydrates thereof, wherein the halogenocyclopropyl group in the formula (X) is a (1R,2S) "2-halogenocyclopropyl groxxp; the aforementioned cosi^^ound, its salts and hydrates thereof,
wherein the halogen atom of the halogenocyclqpropyl group in the
formula (I) is a fluorine atoon;
the aforementioned confound, its salts and hydrates thereof,
wherein the catr^>ound of formula (I) is a stereochemically pure
con^pound;
the aforeinentioned compound; its salts and hydrates thereof,
wherein n in the formula (I) is 1;
the aforementioned cotipound; its salts and hydrates thereof,
wherein the compound of formula (I) is a stereochemically pure
conyound;
7- [3- [1- (S) -amino-l"Cyclqprqpyl]inethylj^rrolidin-l-yl] "6-fluoro-
1- [2- (S) -f luoro-l- (R) -cycloprqpyl] -1,4-dihydro-8-methoxy-4-
oxoquinoline-3-carboxylic acid, its salts and hydrates thereof;
5-amino-7- [3- [1- (S) -amino-l-cyclopropyl]methylpyrrolidin-l-yl] -
6-f luorO"l- [2- (S) -f luoro-1- (R) -qycloprppyl] -1,4-dihydro-8-
roethyl-4-oxocpiinoline-3-carbo3ylic acid, its salts and hydrates
thereof;
S-amino-?- [3- [1- (S) -amino-l-cyclppropyl]methylpyrrolidin-l-yl] -
6,8-dif luoro-1- [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-4-
oxoquinoline-3-carboxylic acid, its salts and hydrates thereof;
the aforementioned con^xjund, its salts and hydrates thereof,
wherein Y is a hydrogen atom;
a drug containing the aforementioned can^x^und, its salts and
hydrates thereof as an active ingredient; and
an antibacterial agent containing the aforementioned ccm^xsund,
its salts and hydrates thereof as an active ingredient.
[Embodizneni: for Carrying out Invention]
Each of the substituents of the coapotmd of the present invention represented by formula (I) :
(wherein R^, R^, n and Q are as defined in the foregoing) will be explained in the following.
The substituents R^ and R^ is each independently a hydrogen atcsn or an alkyl groiip having 1 to 6 carbon atoms, wherein the alkyl grot^ zaay have one or more substituents selected from the group consisting of a hydroxyl gror^, a halogen atom, an alkylthio group having 1 to 6 carbon atoms and an alkylo3cy group.
The alkyl group may be either straight or branched group having 1 to 6 carbon atoms, and the preferred exanples thereof are methyl, ethyl, normal, propyl and isqprqpyl groups-
When the alkyl grovp has a hydroscyl group as a substituent, the alkyl group may be either straight or branched form having 1 to 6 carbon atoms, and the hydroxyl group may preferably be substituted on the terminal carbon atom of the alkyl group. Preferred exaxnples of the all^l group having a
hydroxy! grot^ inclxide those which have 1 to 3 carbon atoms, such as a hydro3^inethyl grot^), a 2-hydro3^ethyl groiqj, a 2-hydro3cyprqpyl groT:^) and a 3-hydro3cypropyl grox:ip.
Vhen the allcyl group has a halogen atom as a sisbstituent, the alkyl group may be either straight or branched form having 1 to 6 carbon atoms, and a fluorine atom is desirable as the halogen atom. With regard to the nuxnber of fluorine atoms, it may be any one of from mono-substitution to perfluoro substitution. The examples thereof are monofluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2-difluoroethyl and 2,2,2-trifluoroethyl groa:ps. Among these groups, a monof luoromethyl group and a 2-f luoroethyl group are more preferred.
When the alkyl group has an all^lthio group as a substituent, the alkyl group may be either straight or branched form having 1 to 6 carbon atoms, and the alkylthio group may also be either straight or branched form having 1 to 6 carbon atoms. An al3ylthiomethyl group, an allcylthioethyl group and an alkylthiopropyl groi:^) are desirable as the alkyl group having an alkyl thio group, and the alkyl thio group may preferably have 1 to 3 carbon atoms. More preferred exanqples are a metJiylthiomethyl group, an ethylthicmethyl group and a methylthioethyl grooqj.
When the alkyl groi:qp has an alkoxyl group as a stibstituent, the alkyl groi^ may be either straight or branched form having 1 to 6 carbon atoms, and the alkoxyl groiip may also
be either stxalght or branched form having 1 to 6 carbon atoms. An alkoxymethyl groi^), an alkosc/ethyl group and an alkoxypropyl groiip are desirable as the alkyl groiip having an alkoxyl group, and the alko^^l grovp may preferably have up to 3 carbon atoms. More preferred examples thereof are a methoxymethyl groi:^, an ethoxymethyl groi^ and a metho^^ethyl group.
The symbol n Is an Integer of from 1 to 4, preferably 1 or 2, and more preferably 1.
Q is a partial structure represented by the following formula (la) .
In the above formula (la) , R^ is an allc^l group having 1 to 6 carbon atoms, an aJLkenyl groTjp having 2 to 6 carbon atoms, a halogenoalkyl group having 1 to 6 carbon atoms, a cyclic alkyl groxap having 3 to 6 carbon atoms which may have a substituent, an aryl groiqj which may have a siabstituent, a heteroaryl group which may have a substituent, an alkoxyl groi:^ having 1 to 6 carbon atoms or an alkylamino group having 1 to 6 carbon atoms.
In this case, an ethyl group is particularly desirable as the alkyl group having 1 to 6 carbon atoms. As the alkenyl grotap having 2 to 6 carbon atoms, a vinyl groixp or a 1-isoprqpenyl gxoup is desirable. A 2-fluoroethyl group is
desirable as the halogenoaJJ^l grotxp having 1 to 6 carfaon^ atoms. A cyclpprppyl grotip is particularly desirable as the cyclic alkyl grot^^ and a halogen atom, particularly a fluorine atcsm, is desirable as the substituent of the cyclic alkyl gror:^.
Exan^les of the aryl grotip which may have a substituent are a phenyl group which may have 1 to 3 substituents selected frcEm the group consisting for exanple of fluorine, chlorine, bromine or the like halogen atom, a hydro^iyl group, an amino grotip, a niliro groiip, an alkyl group having 1 to 6 carbon atoms and an alkoxyl groixp having 1 to 6 carbon atoms, and its preferred illustrative examples are a phenyl group, a 2-fluorophenyl group, a 4-fluorophenyl grotrp, a 2,4-dif luorophenyl grotcp, a 2-fluoro-4-hydroxyphenyl group, a 3-amino-4,6-difluorophenyl group and a 4,6-difluoro-3-methylaminophenyl groiip.
The heteroaryl group is a coa:^>ound derived from a five-or six-xnembered aromatic heterocyclic compound which contains one or more hetero atoms selected from a nitrogen atom, an oxygen atom and a sulfur atom. Exanples thereof are a pyridyl group and a pyrimidyl groa:^). As the substituent on these rings, an alkyl group, a halogen atom or the like is desirable. Particularly preferred is a 5"amino-2,4-difluoropyridyl group,
A methoxyl grotip is desirable as the alkoxyl grot^) having 1 to 6 carbon atoms. A methylamino grotrp is desirable as the al3cylamino grot^ having 1 to 6 carbon atoms.
As the substituent R^, a cyclic alJqrl group or a
halogenocYcloalkyl grorip is desirable. Among these groi^ss, a ^clppropyl groi^ or a 2-halogenocycloprGpYl groizp is particularly desirable. As the halogen atom, a fluorine atom is desirable.
The substituent R^ is a hydrogen atom or an allcylthio grotip having 1 to 6 carbon atcsms, or R^ and R^ maY together form a ring structure by incorporating a paurt of the mother skeleton (namely by including the nitrogen atom to which R*^ is bonded and the carbon atom to which R^ is bonded) . The thus formed ring may contain a sulftir atom as its constituting atom, and the ring may further have an all^l group having 1 to 6 carbon atoms as a substituent. The ring to be formed herein may have a size of from four-membered ring to six-membered ring, and the ring may be saturated or unsaturated.
The substituent X^ is a halogen atom or a hydrogen atom, and a fluorine atom is desirable in the case of the halogen atom. Among these atoms, a fluorine atom or a hydrogen atom is desirable as the substituent.
The substituent R^ is a hydrogen atom, an amino grau^, a hydroxyl groiip, a thiol groigj, a halogenomethyl gravp, an allcyl groiq:» having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl groixp having 2 to 6 carbon atoms ox an alkosiyl group having 1 to 6 carbon atoms, wherein the amino groxrp may have one or two substituents selected from the groiip consisting of a formyl group, an alkyl groiip having 1 to 6 carbon atoms and an acyl group having 2 to 6 carbon atoms.
The alkYl grorp may be either straight or branched groi:^ having 1 to 6 carbon atoms, and its preferred exasples are a methyl grooms, an ethyl grovp, a normal propyl groiip and an isopropyl grot^j. The alkenyl grotip may be either straight or branched groi^ having 2 to 6 carbon atoms and is preferably a vinyl group. The alkynyl grot^) may be either straight or branched group having 2 to 6 carbon atoms and is preferably an ethynyl groi^J. A fluorine atom is particularly desirable as the halogen of the halogenomethyl group, and its number may be from 1 to 3. The alko^c^l groi:^ may have 1 to 6 carbon atoms and is preferably a metho^l grovp.
The substituent R^ is preferably a hydrogen atom, an alkyl group or an amino group, of which a methyl grotp or an unsubstituted amino groiip is more preferred.
When the substituent R^ is an amino group, a hydroa^l groijp or a thiol group, these groups may be protected with ordinally used protective groi:ps.
Examples of such protective groups include tert-butoxycarbonyl, 2,2,2-trichloroetho3(ycarbonyl and the like alkoxycarbonyl groups, benzyloj^carbonyl, para-xnethoxybenzyloxycarbonyl, para-nitrobenzyloacycarbonyl and the like araIkyloxycarbonyl groiips, acetyl, methoxyacetyl, trif luoroacetyl, chloroacetyl, pivaloyl, forrayl, benzoyl and the like acyl groups, tert-butyl, benzyl, para-nitrobenzyl, para-methoxybenzyl, triphenylraethyl and the like alkyl or aral3^1 grotps, methoxymethyl, tert-butoxymethyl, tetrahydropyranyl,
2,2,2*trichloroetho3r^niethYl and the like ethers and trimethylsilyl, isqpropyldimethylsilyl, tert-butyldi metlylsilyl, tribenzylsilyl, tert-butyldiphenylsilyl and the like substituted silyl grovqps, Cmnpounds whose substituents are protected with these protective grot^s are particularly useful as production intermediates.
Wien A^ is a partial structure represented by formula (II):
X^ is a hydrogen atom, an amino groi^, a halogen atcm, a cyano group, a heLlogencmethyl groiip, a halogenomethos^l group, an alkyl grot:^ having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms or an alkoa^l grox^ having 1 to 6 carbon atoms, wherein the amino group may have one or two substituents selected from the group consisting of a formyl group, an all^l groi:^ having 1 to 6 carbon atoms and an acyl group having 2 to 5 carbon atoms.
The allcyl group may be either straight or branched groi:ip having 1 to 6 carbon atoms and the preferred exanples thereof are a methyl grot:^?, an ethyl grorp, a normal propyl group and an isopropyl groi^). The alkenyl group may be either strcuLght or branched group having 2 to 6 carbon atoms and is preferably a vinyl grot^j. The alkynyl grotqj may be either straight or branched group having 2 to 6 carbon atoms and is preferably an
ethynyl grotip. A fluorine atom is particularly desirable as the halogen of the halogenomethyl group, and its number may be from 1 to 3. The alkoxyl group may have 1 to 6 carbon atoms and is preferably methoi^l groi:^^. A fluorine atom is particularly desirable as the halogen of the halogenomethoxyl group, and its number may be from 1 to 3.
Among these substituents; a halogen atom, an allcyl group or an alkoxyl group is desirable^ and a fluorine atom, a methyl grotp or a methoxyl grotxp is more desirable. These substituents are particularly desirable in the case where Q is the partial structure represented by the formula (la) .
In addition, X^ and the aforementioned R^ may together form a hydrocarbon ring structure (size of the ring may be from four-membered ring to seven-membered ring, and the ring may be saturated or unsaturated) by incorporating a part of the mother skeleton (namely by including the carbon atom to which X^ is bonded and the nitrogen atom to which R^ is bonded) , and the thus formed ring may contain an oxygen atom, a nitrogen atom or a sulfur atom as its constituting atom, and the ring may also have an allcyl groiip having 1 to 6 carbon atoms as a^ substituent.
The partial structure represented by the aforementioned fom:ula (la) is desirable as Q, In this case, it is desirable that A^ is the partial structure of formula (II) .
When Q is the partial structure of formula (la) and A^ is the partial structure of the formula (II) , a preferred combination of R^ and X^ is a case in which R^ is an amino group.
a hydrogen a,tom, a hydrosc^l group or an alkyl grox:^ having 1 -to 6 carbon atoms and X^ is a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkoxyl grotip having 1 to 6 carbon atoms, a halogenomethoxyl group or a hydrogen atom.
A more preferred combination is a case in which R^ is an amino groiip, a hydrogen atom, a hydrosqfl group or a methyl group and X^ is a fluorine atom, a methyl grot^, a methoxyl groiip, a difluoromethoxyl group or a hydrogen atom.
A most preferred combination is a case in which R^ is an amino group, a hydrogen atom, a hydroxyl group or a methyl group and X" is a fluorine atom, a methyl group or a methoxyl group. For these R^ and X^ groups, a fluorine atom is desirable as X^. When the substituents X^ and X^ are halogen atoms, X^ is particularly preferably a fluorine atom and X^ is preferably a fluorine atom or a chlorine atom.
Next, the halogenocycloprppyl grot^^ of R^ will be explained.
As the substitutable halogen atom, a fluorine atom and a chlorine atom can be exemplified, of which a fluorine atom is particularly preferred.
Regarding the steric environment at this moiety, it is particularly desirable that the halogen atom and pyridonecarboxylic acid moiety take cis-configuration on the cyclopropane ring.
So-called enantiomorphic isomers are present due to the cis-2-halogenocycloprppyl moiety alone of R"^, and strong
antibacterial activity and high safety have been found in both
isomers.
The compoiand of the present invention shows excellent
characteristics by having a soabstituent represented by the following fornnala at the 10-position of the 2,3-dihydro-3-(S)-methyl-7-oxo-7H-pyrido [1,2,3-de] [1.4]benzoxazine-6-carbo3(ylic acid skeleton or at the 7-position of the 6-fluoro-l-[2-(S)-f luoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid skeleton.
This stibstituent exists in the following four optical isomeric forms, due to the asymmetric carbon atom at 3-position of the pyrrolidine ring and the asymmetric carbon atom at 1-pcsition of the cycloalkyl-substituted aminomethyl substituent.
Among these, the present Inventors considered that the structure of the following fonttula was more desirable.
That is, it was revealed that, when the 10-position of the 2,3-dihydro-3- (S) -inethyl-7-oxo-7H-pyrido [1,2,3-de] [1.4]benzoxazine-6-carboxylic acid skeleton or the 7-position of the 6-fluoro-l-[2-(S)-fluoro-l-(R)-cyclopropyl]-l,4-dihydro-4-oxoquinoline-3-carboxylic acid skeleton has the soabstituent represented by the above forzmila, the coQ^>ound of the present invention shows strong antibacterial activity lapon Gram-negative bacteria and Gram-^positive bacteria and also shows excellent safety and good pharmacokinetics, such as negativness of the micronuclear test (very weak micronucleus induction toxicity) which was not expected before the present invention.
H^ere the contpound of formula (I) of the present invention has a structure allowing the existence of diastereomers, it is desirable to administer a compound cornprising a pure diastereomer in administration to humans or animals. The term "con^^rising a pure diastereomer" as used herein means not only a case in which it is caanpletely free from the other diastereomer (s) but also a case in which it is in a cheanically pure degree. In other words, it is interpretable
that the other diastereomer (s) may be present: in such a degree that i-t does not exert influences upon physical constan'ts and physiological activities of the con^xDund,
Also, the teroi ^^stereochonically pure' as used herein means a con^x^und consistdJig of one of its stereoiomers when the cai[i$)oxmd has a plurality of isomers due to asymmeiiric carbon atQm(s) contained therein. The term ^^pure" in this case can also be considered in the same manner as described above.
The coaipound of the present invention may be used either in its free form or as an acid addition salt or a salt of its carbosyl group. Exanples of the acid addition salt include hydrochloride, sulfate, nitrate, hydrobromide, hydroiodide, phosphate and the like inorganic acid salts, or methanesulfonate, benzenesulfonate; toluenesulfonate (sulfonate), acetate, citrate, maleate, fumarate, lactate (carboxylate) and the like organic acid salts.
The salt of carboxyl group may be either inorganic or organic salt, and its illustrative exanyles include lithium salt, sodium salt, potassium salt and the like alkali metal salts, magnesium salt, calcium salt and the like alkaline earth metal salts, ammonium salt, or triethylamine salt, N-methylglucamine salt, tris-(hydroxylmethyl)aminoaiethane salt and the like.
Also, these free form, acid addition salts and salts of carboxyl group of the conipound may be present as hydrates.
When the compound of the present invention is used for cuitibacterial purpose, it is desirable to use a carboxylic acid
CG8X^x>und in which the group Y is a hydrogen atom, i^iiile a quinolone derivative whose narhoxylic acid moiety is an ester is useful as a synthesis intermediate or a prodrug. For example, alkyl esters, benzyl esters, alkoxyallcyl esters, phenylalkyl esters and phenyl esters are useful as synthesis intermediates.
Also, the ester to be used as a prodrug is an ester which is susceptible to an in vivo cleavage to form a free carboaylic acid, and its illustrative exanples include acetoxymethyl ester, pivaloyloxymethyl ester, ethoaqfcarbonyl ester, choline ester, dimethylaminoethyl ester, 5-indanyl ester, phthalidinyl ester, 5"al]cyl-2-oxo-l,3-dioxol-4-ylmethyl ester, and oxoall^l ester such as 3-acetoxy-2-ox6butyl eater.
The ccanpouad of the present invention represented hy the formula (I) can be produced by various method, and, in a preferred example of these methods, it can be produc^Ki for exairple by reacting a coii5>oiand represented by formula (III) :
[wherein X^ is a siabstituent which functions as a leaving groirp, such as a fluorine atom, a chlorine atom, a bromine atom, a substituted or unsubstituted phenylsulfonyl group or a substituted or unsubstituted alkylsulfonyl group having 1 to 3
carbon atoms^
Y^ is the y defined in the formula (I) or a boron-containing
groTjp represented by formula (IV) :
(wherein Y^ and Y^ each represents a fluorine atom or an allcylcarbonyloxy grovqp having 2 to 4 carbon atoms) , and R^, R^, R^, A^ and X^ are as defined in the formula (I)] with a con^xaund represented by formula (V) :
[wherein R^""" and R^"^ each independently represents a hydrogen atom, an aUcyl grotip having 1 to 6 carbon atoms or a protective group for amino grovqp, in which the alkyl group may have a stabstituent selected from the group consisting of a hydroxyl group, a halogen atom, an alkylthio groiip having 1 to 6 carbon atoms and an alkoxyl grotip having 1 to 6 carbon atoms, and n is as defined in the formula (I) ] or an addition salt thereof
(ejcaircples of the acid addition salt include hydrochloride, sulfate, nitrate, hydrobroroide, hydroiodide, phosphate and the like inorganic acid salts, or methanesulfonate, benzenesulfonate,
toluenesulfonate (sulfonate) , acetate^ cdtrate, znaleate, foTznarate, lactate (cazboscjflate) and the like organic acid salts) .
The reaction can be carried out using or without using a solvent. The solvent to be used in the reaction may be any solvent which is inert under the reaction conditions, and its illustrative exanples include dimethyl sulfoxide, pyridine, acetoiaitrile, ethanol, chloroform, di methylformaniidR, dimethylacetamide, N-methylpyrrolidone, tetrahydrofuran, water and 3-inethoxybutanol or a mixture thereof.
Preferably, the reaction may be carried out in the presence of an acid acceptor such as an inorganic base (e.g., an alkali metal or alkaline earth metal carbonate or bicarbonate) or an organic base (e.g., triethylamine, pyridine, 1,8-diazabicycloundecene) .
The reaction can be carried out at a temperature of frcan room ten^serature to 200°C, preferably from 25 to 150°C. The reaction is carried out for a period of from 30 minutes to 48 hotxrs and conyletes generally after about 30 minutes to 2 hours.
^4hen the amino group is protected, examples of the protective group for amino grotip include those which are generally used dLn this field, such as tert-butoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and the like alkoxycarbonyl groups, benzyloxycarbonyl, para-methoxybenzyloxycarbonyl, para-nitrobenzyloxycarbonyl and the like ara 1 kyloaynarbonyl groups, acetyl, methoxyacetyl, trifluoroacetyl, chloroacetyl, pivaloyl, fonnyl, benzoyl and the like acyl grot^ss, tert-butyl, benzyl.
para-nitxobenzyl, para-methoxybenzyl, triphenylmethyl and the like all^l or aralkyl groups, methoxymethyl, tert-butoxymethyl;. tetrahydropyraiayl, 2,2,2-txichloroetho3cyinethyl and the like ethers and trimethylsilyl, isqpropyldimethylsilyl, tert-butyldimethylsilyl, tribenzylsilyl, tert-butyldiphenylsilyl and the like siibstituted silyl groups.
When Y and Y^ are an aUcyl group having 1 to 6 carbon atoms, an alkoxymethyl groiip having 2 to 7 carbon atoms or a phenylalkyl groi^ con^xi^sed of an alkylene grot^ having 1 to 6 carbon atoms and phenyl group, the ccii^>ound of interest can be converted into its corresponding carboxylic acid compound by treating it under an acidic or basic condition which is generally employed for the hydrolysis of carboxylic acid esters-When Y^ is a structure of the formula (IV) , its conversion into corresponding carboxylic acid cao^>ox2nd can be effected by allowing the compound (III) to react with the compound (V) and then treating it under an acidic or basic condition.
In addition, when de-protection is necessary, the coitpound of interest represented by the formula (I) can be obtained by removing the protective group under suitable conditions for the protective groxzp.
The compound of formula (V) can be produced by various methods^ and, though not particularly limited, it can be synthesized by a method shown in the reference examples as a preferred example in which synthesis of 3"[1-(S)-amino-1-ci'-cloalkyllmethylpyrrolidine is described as a synthetic example
of 3-[l-amino-l-cycloalkyl]inethylpyxrolidine, so that the costpound of fozxoula (V) can be produced in accordance with this method using a Icnown optically active cycloalk^l-stibstituted glycine derivative.
The cis -2 -f luorocyclopropylaTnine ccoprised of a pure isomer^ which is desirable for the synthesis of the coizpound of formula (I) coanprised of a pure isomer, can be synthesized for exanple by the method described in JP-A-2-231475 (the term ^^JP-A' as used herein means an ^^unexami ned published Japanese patent application") . Synthesis of the conpound of fozmula (I) ccaiprised of a pure isomer can be carried out using the thus obtained optically active cis-2-fluorocyclqprqpylamine derivative as the material, in accordance with the method described for example in JP-A-2-231475 .
The following can be cited as illustrative examples of the ccnpound of the present invention,
10- [3- (R) - [1- (S) -Amino-l-cycloprqpyl]methylpyrrolidine-l-yl] -9-f luoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de] [1.4]benzoxazine-6-narbo3cylic acid;
8-aniino-lO- [3- (R) - [1- (S) -amino-l-cyclopropyl]methylpyrrolidine-1-yl] -9-fluoro-2,3-dihydro-3- (S) --methyl-7-oxo-7H-pyrido [1,2., 3-de] [l,4]benzoxazine-6-carboxylic acid;
7- [3- (R) - [1- (S) -amino-l-cyclqpropyl]methylpyrrolidin-1-yl] -6-f luoro-1- [2- (S) -f luoro-1- (R) -cycloprppyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carhoxylic acid; 5-amino-7- [3- (R) - [1- (S) -amino-l-cyclopropyl]methylpyrrolidin-l-
I
yl] -6-fluoro-1- [2- (S) -fluoro-1- (R) -cyclppropyl] -1,4-dihydro-8-inetho3^-4-oxoquinoline-3-carbo3cylic acid;
7- [3- (R) - [1- (S) "amino-l"cycloprqpyl]inethylpyrrolidin-"l"yl] -6-fluoro-1- [2- (S) -fluoro-1- (R) -qyclpproipyl] -1,4-dihydro-8-niethyl-4-oxoquinoline-3-carbcaylie acid;
5-ainino-7- [3- (R) - [1- (S) -amino-l-cyclopropyl]inethylpyrrolidin-1-yl] -6,8-difluoro-1- [2- (S) -fluoro-1- (R) -cyclppropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid; and 5-amino-7- [3- (R) - [1- (S) -cyclppropyl-1-N-
inethylainino]methylpyrrolidin-1-yl] -6,8-difluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid. These compounds have the following structures.
Since the cosipound of the present invention has strong antibacterial actions, it can be used as drugs for use in human bodies^ animals and fishes or as preservatives of agricultural chsnicals and food.
When the coo^x^und of the present invention is used as a drug for human bodies, its dose is within the range of from 50 mg to 1 g, preferably from 100 mg to 300 mg, per day for an adult.
Its dose as a drug for use in animals varies depending on the purpose of its administration (treatment or prevention) , kind and size of each animal to be treated and kind and degree of each infected pathogenic bacterium, but the dose may be within the range of generally from 1 mg to 200 mg, preferably from 5 mg to 100 mg, per 1 kg body weight per day.
The daily dose may be used once a day or by dividing it into 2 to 4 doses per day. As occasion demands, the daily dose may exceed the aforementioned range.
Since the compound of the present invention is active agai.nst a broad range of microorganisms causing various infectious diseases and effective to treat, prevent or alleviate
diseases induced by these pathogens.
Illustrative examples of bacteria and bacterioid microorganisms on which the coinpotand of the present invention is effective include those which belong to the genus Staphylococcus, Streptococcus pyog&as, hemolytic streptococci, enterococcus, pneumococcus / those which belong to the genus Peptostreptococcus, Neisseria, gonorrhoeae, Escherichia coll, those which belong to the genera Cltrobacter and Shigella, Klebsiella pneumoniae, those which belong to the genera Enterobacter, Serratla and Prot&us, Fseudamonas aeruginosa, Haemophilus influenzae, those which belong to the genera Acinetobacter and CanpyldbactBr and Chlamydia trachomatis. Illustrative examples of diseases which are induced by these pathogens include folliculitis, furuncle, carbuncle, erysipelas, phlegmon, lysphangitis/lymphadenitis, felon, subcutaneous abscess, hidradenitis, acne conglobata, infectious atheroma, perirectal abscess, mastitis, superficial secondary infections after injury, bum injury, operative wound and the like, pharyngitis, acute bronchitis, tonsilitis, chroiaic bronchitis, bronchiectasis, diffuse bronchiolitis, secondary infection of chronic respiratory disease, pneumonia, pyelonephritis, cystitis, prostatitis, epididymitis, gonococcal urethritis, nonspecific urethritis, cholecystitis, cholangitis, bacillary dysentery, enteritis, uterine adnexitis, intrauterine infection, bcurtholinitis, blepharitis, hordeolum, dacryocystitis, tarsadenitis, corneal ulcer, octitis media, sinusitis,
periodentitis, pericoronitis, jaw infection, peritonitis, endocarditis, sepsis, meningitis and skin infection.
The contpound of the present invention is also effective against various microorganisms causing dLnfectious diseases in animals, such as those which belong to the genera EscbBrlcbla, SaJjQonella, Pastefiurella, HBesnophllus, BordBtella,, StapbYlococcus and Myxzaplasma,. Illustrative examples of such diseases include colxbacillosis, pullorum disease, avian paratyphoid, avian cholera, infectious coryza, staphylococcosis, nrycoplasma infection and the like in the case of birds; colibacillosis, salmonellosis, pasteurellosis, haemophilus infection, atrophic rhinitis, exudative epidermis, mycoplasma infection and the like in the case of pigs; colibacillosis, salmonellosis, hemorrhagic sepsis, inycoplasma infection, bovine pleuropneumonia, bovine mastitis and the like in the case of cattle; colisepsis, salmonella infection, hemorrhagic sepsis, uterine empyema, cystitis and the like in the case of dogs; and exudative pleurisy, cystitis, chronic rhinitis, haemophilus infection, kitten diarrhea, mycoplasma infection and the like in the case of cats.
The antibacterial preparation which conprises the compound of the present invention can be prepared by selecting an appropriate preparation depending on each administration method and employing generally used various preparation method. Recjaxding the dosage form of the antibacterial preparation which uses the compound of the present invention as its principal
agezi't, tablets, powders, granules, c^^sules, solutions, syrups, elixirs, oily or aqueous suspensions and the like can be exemplified as oral preparations.
Regarding injections, a stabilizing agent, an antiseptic agent and a solubilizing agent may be used in the preparation, or a solution which may contain these auxiliary agents may be contained in a container and made into a solid preparation by freeze-drying or the like means to be re-dissolved when used. In addition, a single dose may be contained in a single container or multiple doses may be contained in the same container.
Also, solutions, suspensions, emulsions, ointments, gels, creams, lotions, sprays and the like can be exenplified as preparations for external use.
Solid preparations may contain pharmaceutically acceptable additives together with the active conpound and can be prepcored for example by mixing the compound with additives optionally selected from fillers, extenders, binders, disintegrators, solubilization enhancing agents, moistening agents, lubricating agents and the like. As liquid preparations, solutions, suspensions, emulsions and the like can be exen^jlified, which may contain a suspending agent, an emulsifying agent and the like as additives.
Exan^^les of the method for administering the cooEpound of the present invention to animals include a method in which it is orally administered directly or by mixing it with feed, a method
in which it is made into a solution and then orally administered directly or by mixing it with drinking water or feed and a method in which it is administered by injection.
Regarding the pharmaceutical preparations for use in the administration of the conpound of the present invention to animals, it can be made optionally into powders, fine subtilaes, sellable powders, syrups, solutions or injections making use of the techniques generally used in this field.
Formulation exaniples of the pharmaceutical preparations are shown below.
BEST M3DE FOR CARRYING OUT INVENTION
Examples of the present invention are given below by way
of illustration and not by way of limitation.
[Reference Example 1] 1 "Cyclopropyl-2 -propen-1 -one
Under a stream of nitrogen ^. cyclopropylroetkyl ketone
(6.33 g, 75.2 mmol) was dissolved in anhydrous tetrahydrofuran
(75 ml) . While stirring under ice-cooling, to this was added dropwise a solution which had been prepared by dissolving N-methylanilinium trifluoroacetate (25.0 g, 113 mmol) in 37% formaldehyde aqueous solution (10.2 ml) under ice-cooling-After the dropwise addition, the reaction solution was heated Tinder reflux for 7 hotirs. After cooling, the reaction solution was mixed with diethyl ether (100 ml) and stirred, and then the organic layer was separated. The aqueous layer was extracted with diethyl ether (50 ml) , The organic layers were coihbined, gradually mixed with saturated sodiimi bicarbonate aqueous solution (100 ml) and stirred, and then the organic layer was separated. The thus separated organic layer was washed with saturated sodium chloride aqueous solution (100 ml) . This was dried over anhydrous magnesium sulfate, filtered and then
concentrated to 8.01 g under a reduced pressiire of 150 mmHg,
thereby obtaining a yellow oil containing the title compound.
This product was used in the subsequent reaction without
purification.
^H-NJdR (400 MHz, CDCI3) 5: 0.90 - 0.96 (2H, m) , 1.08 - 1,13 (2H, m), 2.14 - 2.25 (IH, m) , 5.82 (IH, dd, J = 10.74, 1.47 Hz), 6.29
(IH, dd, J = 17,57, 1.47 Hz), 6.47 (IH, dd, J = 17.57, 10.74 Hz)
[Reference Exantple 2] Cyclopropyl [1- [1- (R) -phenylethyl]pyrrolidin-3-yl] ketone
The product containing l-cyclopropyl-2-propen-l-one described in Reference Exainple 1 (8.01 g) and N-(normal-butC3cyinethyl) -N- [1- (R) -phenylethyl] trimethylsilylmethylamine
(23.2 g, 79.9 xnmol) were dissolved dLn dry dichloroethane (350 ml) to which was subsequently added dropwise trifluoroacetic acid (500 (jJ.) . After 12 hours of stirring at room tenperature, the reaction solution was washed with saturated sodium bicarbonate aqueous solution (100 ml) and saturated sodium chloride aqueous solution (100 ml) in that order. This was dried over anhydrous magnesiimi sulfate, filtered and then concentrated under a reduced pressure. The resulting residue was applied to a flash silica gel chromatography and eluted with n-"h
Hz), 1,62 - 1.76 (IH, m), 1.90 - 2.17 (2H, m) , 2.35 - 2.93 (4H, m) , 3.22 - 3.26 (2H, m), 7.23 - 7,34 (5H, m) [Reference Example 3] 3- [l- (tert-Butoscycarhonyl) ami no-1-c;fGloprqpyl]methyl"l- [1- (R) -phenylethyl]pyrrolidine
Cyclopropyl [1- [1- (R) -phenylethyl]pyrrolidin-3^1] ketone (1.563 g, 7.793 mmol) was dissolved in anhydrous methanol (25 ml) . To this were added ammonitim acetate (5.236 g, 67.93 mmol) , sodium cyanoborohydride (435.2 mg, 6.925 mmol) and Molecular Sieves 4A powder (1.86 g) , and the mixture was stirred at room tismperature for 16 hours tonder a stream of nitrogen. The reaction solution was filtered through celite, and the solvent was ev^>orated under a reduced pressvire. The resultdLng residue was dissolved in dichloromethane (100 ml) , and the solution was washed with saturated sodium bicarbonate aqueous solution (50 ml) and saturated sodium chloride aqueous solution (50 ml) in that order^ and then dried over anhydrous magnesium sulfate. After filtration; the solvent was concentrated under a reduced pressure. The thus obtained residue was dissolved in dry dichloromethane (25 ml) to which, under ice-cooling, was subsequently added dropwise dichloromethane (5 ml) solution of di"tert-butyl bicarbonate (2,225 g, 10,19 mmol) , The reaction solution was stirred at room tenperature for 2 hours and then concentrated under a reduced pressure. The thus obtained residue was applied to a flash silica gel chromatography and eluted with chloroform;methanol = 10:1, thereby obtaining 1.299 g (55.5%) of the title con^Joxind as a colorless oil. In this
connection^ this product was obtained as a zoixture of four optical isomers.
^H-NMR (400 MHz, CDCI3) 6: 0,20 - 0.30, 0.35 - 0,52, 0,68 - 0.78 (4H, m), 1,36 (3H x 1/4, d, J = 5.86 Hz), 1.39 (3H x 3/4, d, J = 5.86 Hz), 1.43 (9H x 1/4, s), 1.45 (9H x 3/4, s), 1.61 - 1.74 (IH, m), 2,25 " 2.76, 2.80 - 3.07, 3.18 - 3.26 {9H, m), 5.28 (IH, brs), 7.23 - 7.34 (5H, m)
[Reference Exarnple 4] l-Benzyloxyc^rbonyl-3"[1-(tert-butoxycarbonyl) amino-l-cyclopropyl] inethylpyrrolidine (Fl, F2, F3 and F4)
3- [1- (tert-Buto3^carbonyl) amino-l-cyclopropyl]methyl-1-[l-(R)-phenylethyl]pyrrolidine (1.234 g, 3.582 mmol) was dissolved in dry dichloromethane (20 ml) to which, under ice-cooling, was subsequently added drqpwise benzyl chloroformate (1278 ^1, 8,955 mmol) . After 8 hours of stirring at room ten;>erature, the reaction solution was concentrated under a reduced pressure. The thus obtained residue was ^3plied to a flash silica gel chromatography and eluted with n-hexane: ethyl acetate = 2:1, thereby obtaindLng 959 mg (71,5%) of the title coit;x>iuid as a colorless oil.
Thereafter, this product was subjected to fractional HPLC using a chiral colomm to isolate and purify foxir optical isomers Fl, F2, F3 and F4. HPLC fractionation conditions;
Column: CHIRALPAKAD (Daicel Chemical Industries) , 2 cm x 25 cm Mobile phase: n-hexane:2-propanol = 80:20 (v/v)
it was confirmed that each combination of Fl and F4 and F2 and F3 has the enantiomorphic relation, [Inventive Exainple 1] 5-Atnino-7- [3- (1-atnino-l-
cyclopropyl)niethylpyrrolidin-l-yl] -6,8-dif luoro-1- [2- (S) -f luoro-1" (R) -cyclopropyl] -1,4-dihydro-4-oxoqainoiine-3-carboxylic acid (the substituent at the 7-position is originated from Fl)
l-Benzylo3cycarbonyl-3- [1- (tert-butoxycarbonyl) amino-1-cyclopropyl]2nethylpyrrolidine (originated from Fl of Reference Exaznple 4; 185 zng, 0.494 mmol) was dissolved in anhydrous methanol (30 ml) , and the solution was mixed with 10% palladitnn on carbon catalyst (water content 50%, 200 mg) smd stirred at room -hemperature for 1 hour in an atmosphere of hydrogen under atmospheric pressure. After filtration of the reaction solution through celite, the resulting filtrate was concentrated under a reduced pressure. The thus obtained residue and triethylamine (2 ml) were added to dry acetonitrile (10 ml) , and the mixture was further mixed with 5-amino-6,7,8-trifluoro-l-[2-(S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-caxboxylic acid (130 mg, 0.412 mmol) and heated under reflux for 16 hours. Aftar cooling of the reaction solution, the thus precipitated
crystals were collected by filtration, washed with acetonitrile, mixed with concentrated hydrochloric acid (10 ml) under ice-cooling and then stirred at room tenperature for 5 minutes. This was mixed with distilled water (15 ml) , and the thus obtained acidic aqueous solution was washed with dichlorosnethane (20 ml X 2) , adjusted to pH 11 with sodium hydroxide aqueous solution under ice-cooling and then washed with chloroform (10 ml) . The resulting basic aqueous solution was adjusted to pH 7.4 with 1 N hydrochloric acid and extracted with chloroform (100 ml X 4) , After drying over anhydrous sodium sulfate, the solvent was evaporated under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing frcsa ethanol-28% aqueous ammonia and then dried under a reduced pressure, thereby obtaining 160 mg (88.9%) of the title coo^x^tind as yellow needle crystals.
^H-NMR (400 MHz, 0,1 N NaOD) 6: 0.11 - 0.18 (2H, m) , 0.39 (2H, d, J= 7.81 Hz), 0.57 - 0.67 (IH, m) , 1.24 - 1.35 (IH, m) , 1.43 -1.61 (3H, m) , 1.93 - 2.06 (2H, m) , 3,20 - 3.26 (IH, m) , 3.37 -3.49 (2H, m) , 3.59 - 3.72 (2H, m) , 4,97 (IH, dm, J = 64.16Hz), 8.21 (IH, s)
Melting point: 185 - 193°C
Elemental analysis data for C2iH23F^403 • 0.25H2O Calcd.: C, 57.20; H, 5.37; N, 12.71 Found : C, 57,16; H, 5.39; N, 12.88 [Inventive Example 2] 5-Anino-7"[3-(1-amino-l-cyclopropyl)methylpyrrolidin-l-yl] -6,8-dif luoro-1- [2- (S) -fluoro-
1- (R) -cyclopropyl] -1,4-dihydro-4"Oxoquinoline-3-caTt>oxylic acid (the siibstituent at. the 7-position is originated from F2)
l-Benzylo3^carbonyl-3- [1- (tert-butoxycarbonyl) amjno-l-cyclopropyl]inethylpyrrolidine (originated from F2 of Reference Exainple 4; 75 zng, 0.200 mmol) was dissolved in anhydrous methanol (15 ml) , and the solution was mixed with 10% palladium on carbon catalyst (water content 50%, 100 mg) and stirred at room ten5>erature for 1 hour in an atmosphere of hydrogen under atmospheric pressure. After filtration of the reaction solution through celite, the resulting filtrate was concentrated under a reduced pressure. The thus obtained residue and triethylamine (1 ml) were added to dry acetonitrile (5 ml) , and the mixture was further mixed with 5-amino-6,7,8-trifluoro-l-[2-(S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid (53 mg, 0.167 mmol) and heated under reflux for 12 hours. After cooling of the reaction solution, the thus precipitated crystals were collected fay filtration, washed with acetonitrile, mixed with concentrated hydrochloric acid (5 ml) under ice-cooling and then stirred at room temperature for 5 minutes. This was mixed with distilled water (10 ml) , and the thus obtained acidic aqueous solution was washed with dichlorcmethane (15 ml x 2) , adjustied to pH 11 with sodium hydroxide aqueous solution under ice-cooling and then washed with chloroform (10 ml) . The resulting basic aqueous solution was adjusted to pH 7,4 with 1 N hydrochloric acid and extracted with chloroform (80 ml x 3) . After drying over anhydrous sodium sulfate, the solvent was
evaporated -under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from ethanol-28% aqueous ammonia and then dried under a reduced pressure, thereby obtaining 55 mg (75.5%) of the title conpound as yellow needle crystals.
^H-NMR (400 MHz, 0.1 N NaOD) 5: 0.13 - 0.19 (IH, m) , 0.22 - 0.30 (IH, m), 0.41 - 0.49 (IH, m) , 0.52 - 0.60 (IH, m) , 0.73 - 0.84 (IH, m), 1.53 - 1.72 (3H, m) , 1.86 (IH, t, J = 9.28 Hz), 2.11 -2.29 (2H, m), 3.44 - 3.57 (2H, m) , 3.62 - 3.68 (IH, m) , 3,72 -3.95 (2H, m), 4.96 (IH, dm, J = 63.95 Hz), 8,24 (IH, s) Melting point: 190 - 192°C
Elemental analysis data for C21H23F3N4O3 • 0.25H20 Calcd.: C, 57.20; H, 5.37; N, 12.71 Found : C, 57.27; H, 5.36; N, 12,65 [Inventive Example 3] 5-Amino-7- [3- (1-amino-l-
cyclopropyl)methylpyrrolldln"l"yl] -6,8-dif luoro-1- [2- (S) -f luoro-1- (R) -cyclqpropyl] -1,4"dlhydro-4-oxoquinoline-3"carfaoxyllc acid (the STjbstituent at the 7-posltion is originated from F3)
l-Benzylo3qfcarbonyl-3- [1- (tert-butoa^carbonyl) amlno-1-cyclopropyl]methylpyrrolidine (originated from F3 of Reference Example 4; 100 mg, 0,267 mmol) was dissolved in. anhydrous methanol (20 ml) , and the solution was mixed with 10% palladium on carbon catalyst (water content 50%, 100 mg) and stirred at room temperature for 1 hour in an atmosphere of hydrogen under atmospheric pressure. After filtration of the reaction solution through celite, the resulting filtrate was concentrated under a
reduced pressure. The thus dbtained residue and trlethylazoine (2 ml) were added to dry acetoxiltrile (8 ml) , and the xnixture was further mixed with 5-amino-6,7,8-trifluoro-1- [2- (S) -fluoro-1- (R) -cycloprqpyl] -1,4"dihydro-4-oxoquinoline-3-carbo3ylic acid (80 mg, 0.222 mmol) and heated under refliox for 16 hours. After cooling of the reaction solution, the thus precipitated crystals were collected by filtration, washed with acetonitrile, mixed with concentrated hydrochloric acid (10 ml) under ice-cooling and then stirred at room tenperature for 5 minutes. This was mixed with distilled water (15 ml) , and the thus obtained acidic aqueous solution was washed with dichloromethane (20 ml x 2) , adjusted to pH 11 with sodium hydroxide aqueous solution under ice-cooling and then washed with chloroform (20 ml) . The resulting basic aqueous solution was adjusted to pH 7.4 with 1 N hydrochloric acid and extracted with chloroform (80 ml x 4) . After drydLng over anhydrous sodium sulfate, the solvent was evaporated under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from ethanol-28% aqueous ammonia and then dried under a reduced pressure, thereby obtaining 59 mg (60,8%) of the title compound as yellow needle crystals.
^H-NMR (400 MHz, 0.1 N NaOD) 5: 0.12 - 0.17 (IH, m) , 0.21 - 0.28 (IH, m), 0.41 - 0.48 (IH, m), 0.51 - 0.60 (IH, m), 0.72 - 0.82 (IH, m) , 1.44 - 1.53 (3H, m), 1.81 (IH, t, J = 8.79 Hz), 2.09 -2.28 (2H, m), 3.41 - 3.47 (IH, m) , 3.49 - 3.57 (IH, m), 3.59 -3.66 (IH, m), 3.72 - 3,80 (2H, m) , 4.95 (IH, dm, J = 65.11 Hz),
8.19 (IH, s)
Melting point: 193 - 194°C
Elemental analysis data for C2iH23F3N4O3-0,25H2O
Calcd. : C, 57.20; H, 5.37; N, 12.71
Found : C, 57.21; H, 5.37; N, 12.70 [Inventive Example 4] 5"Amino-7"[3-(l-amino-l-
cycloprqpyl)inethylpyrrolidin"l"yl] -6,8-dif luoro-1- [2- (S) -f luoro-1- (R) -cycloprqpyl] -1,4-dihydro"4*-oxoquinoline"3"carhoxylic acid (the substituent at the 7-position is originated from F4)
l-Benzylo3cycarbonyl"3- [1- (ter t-bu to3ycarbonyl) amino-1-cycloprqpyl]inethylpyrrolidine (originated from F4 of Reference Exanple 4; 200 mg, 0.534 znznol) was dissolved in anhydrous loethanol (20 ml) , and the solution was mixed with 10% palladinm on carbon catalyst (water content 50%, 200 mg) and stirred at room t.eniperature for 1 hour in an atmosphere of hydrogen under atmospheric pressure. After filtration of the reaction solution through celite, the resulting filtrate was concentrated under a reduced pressure« The thus obtained residue and triethylamine (3 ml) were added to dry acetonitrile (15 ml) , and the mixture was further mixed with 5-amino-6,7,8-trifluoro-l-[2-(S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid (141 mg, 0.445 mmol) and heated under reflux for 16 hours. After cooling of the reaction solution, the thus precipitated crystals were collected by filtration, washed with acetonitrile, mixed with concentrated hydrochloric acid (10 ml) under ice-cooling and then stirred at rocon t^emperature for 5 minutes.
This was mixed with dis-tilled water (20 ml) , and the thus obtained acidic aqueous solution was washed with dichlorcmethane (20 ml X 2) / adjusted to pH 11 with sodium hydroxide acjueous solution under ice-cooling and then washed with chloroform (20 ml) . The resulting basic aqueous solution was adjusted to pH 7.4 with 1 N hydrochloric acid and extracted with chlorofoiaa (100 ml X 4) . After drying over anhydrous sodium sulfate, the solvent was evaporated under a reduced pressure. Thereafter, the resulting residue was, purified by recrystallizing from ethanol-28% aqueous ammonia and then dried under a reduced
pressoire, thereby obtaining 138 mg (71.1%) of the title compound as yellow needle crystals.
^H"NMR (400 3^z, 0.1 N NaOD) 5: 0.12 - 0,23 (2H, m) , 0.41 - 0.51 (2H, m), 0.70 - 0.87 (IH, m) , 1.46 - 1.66 (3H, m) , 1.80 - 1.88 (IH, m) , 2.08 - 2.30 (2H, m) , 3.46 - 3.60 (2H, m) , 3.62 - 3.69 (IH, m) , 3.72 - 3.82 (2H, m) , 4.96 (IH, dm, J = 63.89 Hz), 8.19 (IH, s)
Melting point: 188 - 194°C
Elemental analysis data for C21H23F3N4O3
Calcd.: C, 57.79; H, 5.31; N, 12.84 FoTond : C, 57.56; H, 5.39; N, 12,88 [Reference Example 5] Ethyl 4- (S) -benzyloxycarbonylamino-4-
cyclopropyl-3-oxobutanoate
Malonic acid monoethyl ester (988.4 mg, 7.482 mmol) was
dissolved in anhydrous tetrahydrofuran (20 ml), and the solution
was mixed with magnesium ethoxide (488 mg, 3.93 mmol) xander ice-
cooling and then s'tirred at room teinperature for 2 hours. The reaction solution was concentrated to dryness under a reduced pressure, and the thus obtained powder was dissolved in anhydrous tetrahydrofuran (30 ml) .
L"N-Benzylo3cycarbonyl-cyclqpropylglycine (1.332 g, 5.334 znmol) was dissolved in anhydrous tetrahydroftiran (20 ml) , and the solution was mixed with N^N^ -carbonyldiimidazole (910 mg, 5.61 mmol) under ice-cooling and then stirred at room tenaperature for 2 hours. Under ice-cooling, the aforementioned tetrahydrofuran solution previously prepared was added dropwise to the just prepared solution, and the mixture was stirred at room temperature for 16 hours.
The reaction solution was concentrated under a reduced pressure, euid the resulting residue was mixed with toluene (60 ml) and 10% citric acid aqueous solution (50 ml) and stirred at room temperature for 5 minutes. The organic layer was separated and the aqueous layer was extracted with toluene (20 ml x 2) . The organic layers were combined, washed with water (50 ml) and saturated sodium chloride aqueous solution (50 ml) in that order, and then dried over anhydrous magnesium sulfate. After filtration, the solvent was evaporated under a reduced pressure and the resulting residue was applied to a flash silica gel chromatography and eluted with n-hexane: ethyl acetate = 2:1, thereby obtaining 1.527 g (89,4%) of the title compound as a colorless oil. ^H-IJMR (400 MHz, CDCI3) 5: 0.53 - 0.61 (3H, m) , 0.67 - 0.77 (IH,
m) , 0.90 - 1.01 (IH, m) , 1,27 (3H, t, J = 7.33 Hz), 3.65 (2H, s) ,
3.89 - 3.93 (IH, m) , 4.20 (2H, q, J = 7.33 Hz), 5.10 (2H, s) ,
5.47 (IH, brs), 7.31 - 7.35 (5H, m)
[Reference Exanrple 6] Ethyl 4-(S) "benzyloxycarbonylamino-4"
cyclopropyl-3-hydro3cyfaii'tanoate
Ethyl 4- (S) -benzyloxycarbonylamino-4-cyclopropyl-3-oxobutanoate (1.526 g, 4.778 mmol) was dissolved in anhydrous ethanol (15 ml) , and the solution was mixed with sodiiim borohydride (94.6 mg, 2.50 mmol)xinder ice-cooling and stirred at the same temperature for 1 hour. Under ice-cooling, the reaction solution was mixed with water (20 ml) and then ethanol was evaporated under a reduced pressure. The thus obtained residue was mixed with chloroform (50 ml) and stirred, and then the thus separated chloroform layer was washed with saturated sodium chloride aqueous solution (30 ml) and dried over anhydrous magnesium sulfate. After filtration, the solvent was evaporated to dryness xinder a reduced pressure, thereby obtaining 1.509 g (98,3%) of the title compound as a colorless oil. This product was used in the subsequent reaction without purification.
>H-NMR (400 MHz, CDCI3) 5: 0.24 - 0.67 (4H, m) , 0,96 - 1.06 and 1.08 - 1.16 (total 1 H, m for each), 1,27 (3H, t, J= 7.32 Hz), 2.45 - 2.58 (IH, m) , 2,66 - 2.73 (IH, m) , 2.84 - 3.01 (IH, m) , 3.33 (IH, brs), 4.17 (2H, q, J = 7.32 Hz), 5.09 (2H, s) , 5.18 and 5.28 (total IH, br-s for each), 7.30 - 7.37 (5H, m) [Reference Exaii5)le 7] Ethyl 4- (S) -benzyloxycarbonylami no-4-
cyclopropyl"2 -butenoate
Ethyl 4- (S) -benzyloxycarfaonylamino"4-cYcloprqpyl"3-hydroxybutanoate (1.488 g, 4.630 irnnol) was dissolved in dry dichlorcmethane (50 ml) and stirred at -15*^C while adding triethylamine (1,291 yH, 9.260 ssnol) , and then methanesulfonyl chloride (449 ^1, 5.80 znmol) was added dropwise thereto and the mixture was stirred at the same tenqperature for 1 hoior. 1,8-Diazabicyclo[5.4.0]-7-undecene (1,486 ^1, 1.955 mmol) was added dropwise to the reaction solution, and the mixture was gradually wanned tip to room tenperature and then stirred for 15 hours. The reaction solution was washed with 10% citric acid aqueous solution (50 ml) , and the organic layer was separated and then the aqueous layer was extracted with chloroform (30 ml) . The organic layers were combined, washed with water (50 ml) and saturated sodium chloride aqueous solution (50 ml) in that order, and then dried over anhydrous magnesiimi sulfate. After filtration, the solvent was evaporated under a reduced pressure, and the thus obtained residue was applied to a flash silica gel cJiromatography and eluted with n-hexane: ethyl acetate = 4:1, thereby obtaining 1.174 g (87,2%) of the title coo5>otmd as a light yellow oil.
^H-NMR (400 MHz, CDCI3) 5: 0.31 - 0,37 (IH, m) , 0,39 - 0,48 (IH, m), 0,52 - 0,65 (IH, m) , 0,86 - 0.95 (IH, m) , 1,29 (3H, t, J = 7.33 Hz), 3.73 (IH, brs) , 4,20 (2H, q, J - 7,33 Hz), 4.92 (IH, brs), 5.11 (2H, s) , 5,97 (IH, d, J = 15.63 Hz), 6,91 (IH, dd, J ^ 15.63, 5.37 Hz), 7.31 - 7,36 (5H, m)
[Reference Example 8] Ethyl 4-(S) "benzyloxycai±>onylainino-4" cycloprc3pyl"3-iu.txomethylbutanoa'te
Ethyl 4- (S) -benzyloxycarbonylamino-4-cyclopropyl-2-butenoate was dissolved in dry nitrcanethane (15 ml) , and the solution was mixed with 1,1,3,3-tetramethylguanidine (133 }il, 1.05 mmol) and stirred at room ten^^erature for 17 hours. The reaction solution was concentrated -under a reduced pressure^ the resulting residue was dissolved in chlorof oma (50 ml) , and the solution was washed with 10% citric acid aqueous solution (50 ml) and saturated soditim chloride aqueous solution (50 ml) in that order, and then dried over anhydrous magnesium sulfate, thereby obtaining 1.207 g (96.1%) of the title coampound as a yellow oil. This product (diastereomer mixture) was used in the subseG[uent reaction without purification.
^H"NMR (400 MHz, CDCI3) 5: 0.31 - 0.45 (IH, m) , 0.48 - 0.56 (IH, m) , 0.61 - 0.70 (IH, m) , 0.82 - 0.92 (IH, m) , 1.26 (3H, t, J = 7.33 Hz) , 2.49 (d, J = 7,32 Hz) , 2.53 (d, J = 6.34 Hz) , 2.67 (d, J =5.72 Hz), 2.71 (d, J = 5.86 Hz) , (2.49, 2.53, 2.67, 2.67, total 1 H) , 3.00 (IH, q, J = 6.34 Hz), 3.13 (IH, q, J = 7.32 Hz), 4.15 (2H, q, J = 7.33 Hz) , 4.57 and 4.59 (total 2 H, d, J = 7.33 Hz), 4.86 (IH, br-s) , 5.10 (2H, s) , 7.31 - 7.36. (5H, m) [Reference Example 9] 4- [1- (S) -tert-Butoxycarbonylamino-1-cyclopropyl]methylpyrrolidin-2-one (isomer A)
Ethyl 4- (S) -benzyloxycarbonylamino-4-cyclopropyl-3-nitromethylbutanoate (16.4 g, 41.0 mmol) was dissolved in ethcuiol (500 ml) , and the solution was mixed with 10% palladium
on carbon catalyst (water content 50%, 16 g) and si:ibjeGted to 5 hours of catalytic hydrogenation at room tignperature. After removal of the catalyst by celite filtration, the resiilting filtrate was heated under reflux for 6 hours. The solvent was evaporated under a reduced pressure, the restilting residue was dissolved in dry dichloromethane, the resulting solution was mixed with triethylamine (8.24 ml, 59.1 mmol) and then with di-tert-butyl bicarbonate (11.32 ml, 49,2 mmol) , and then the mixture was stirred at room ten^^erature for 6 hours. After concentration of the reaction solution under a reduced pressure, the thus obtained residue was applied to a flash silica gel chromatography and eluted with chloroform'.methanol = 95:5, and the thus obtained crystals were purified by recrystallizing them from a chloroform-n-hexane system, thereby obtaining 3.34 g (32.0%) of one diastereomer of the title cosipound as a single ccBipound (isomer) in the foim of white crystals. ^H"NMR (400 MHz, CDCI3) 5: 0.34 - 0.35 (2H, m) , 0.44 - 0.48 (IH, m), 0.58 - 0.65 (IH, m), 0.72 - 0.79 (IH, m), 1.44 (9H, s), 2,28 (IH, dd, J = 17.09, 9.04 Hz), 2.44 (IH, dd, J =17.09, 8.44 Hz), 2.67 - 2.73 (IH, m) , 3.04 - 3.06 (IH, m) , 3.25 - 3.30 (IH, m) , 3.47 (IH, t, J =8.79 Hz), 4.57 (IH, s) , 5.84 (IH, s) [Reference Example 10] l"Benzyl"4-[1-(S)-tert-butoxycarbonylami no-1-cyclopropyl ] methylpyrrolidin-2-one (isomer
A).
Under a stream of nitrogen, 4-[l-(S)-tert-butoxycarbonylamino-l-cyclopropyl]methylpyrrolidin-2-one (3,15 g.
12.4 mmol) was dissolved in dry rJ-iTn^a-n^yi-fnrmsim'^H
^H-NMR (400 MHz, CDCI3) 6: 0.27 - 0.28 (2H, m) , 0.39 - 0.43 (IH, m), 0.52 - 0.56 (IH, m), 0.66 - 0.72 (IH, m), 1.41 (9H, s), 2.39 - 2.42 (IH, m), 2.54 - 2.62 (2H, m), 3,01 (IH, s), 3.29 - 3.33 (IH, m) , 4,40 (IH, s) , 4,33, 4.55 (each 1 H, ABq, J = 14,4 Hz), 7.23 - 7.36 (5H, m)
[Reference Example 11] l-Benzyl-3-[l-(S)-tert-butoxycarbonylamino-l-cyclppropyl]methylpyrrolidine (isomer A)
Under a stream of nitrogen, l-benzyl-4-[l-(S)-tert-butoxycarbonylami no-l-cycloprqpyl]methylpyrrolidin-2-one (2.74 g, 7,95 mmol) was dissolved in dry tetrahydrofuran (70 ml) to which, under ice-cooling, was subsequently added dropwise a borane-tetrahydrofuran conplex (1.0 N tetrahydrofuran solution; 47,7 ml,
47.7 mtnol) . After completion of the dropwise addition^ and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under a reduced pressure, mixed with a solution of ethanol: water = 10:1 (130 ml) and triethylamine (20 ml) , and then heated under reflux for 4 hours. The reaction solution was concentrated under a reduced pressure and mixed with chloroform (100 ml) , and the thus separated organic layer was washed with water (100 ml) and saturated sodium chloride aqueous solution (50 ml) in that order, and then dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under a reduced presstire, and the thus obtained residue was ^splied to a flash silica gel chromatography and eluted with chloroform:methanol = 95:5, thereby obtaining 2,63 g (100%) of the title compound as a colorless oil.
^H"NMR (400 MHz, CDCI3) 5: 0.19 - 0.23 (IH, m) , 0.35 - 0,44 (3H, m), 0.73 - 0.76 (IH, m), 1.44 (9H, s), 1,60 - 1,66 (IH, m) , 1,94 - 2.00 (IH, m), 2,36 - 2.42 (2H, m), 2,51 - 2,66 (3H, m) , 2.95 -3.02 (IH, m) , 3,52, 3.67 (each 1 H, ABq, J = 12.9 Hz), 4.58 (IH, s), 7.24 - 7.34 (5H, m)
[Reference Example 12] l-Benzyloxycarbonyl-3- [1- (S) -tert-butoyycarfaonylaTnino-l-cyclopropyl]methylpyrrolidine (isomer A = isomer F3 of Reference Example 3)
l-Benzyl-3- [1- (S) -tert-butoxycarbonylamino-1-cyclopropyl]methylpyrrolidine (isomer A; 238 mg, 0.720 xnmol) was dissolved in dry dichloromethane (10 ml) to which was
stibsequently added drppwlse benzyl chloroforzoate (309 |il, 2.16 mmol) under ice-ccxDling. After 8 hours of stirring at room temperature, the reaction solution was concentrated under a reduced pressure. The resulting residue was c^splied to a flash silica gel chrcmatography and eluted with n-hexane: ethyl acetate = 2:1, thereby obtaining 197 ing (73.1%) of the title con?x>und as colorless amorphous.
The Rf value of TLC (thin layer chromatography, development with n-hexane: ethyl acetate = 1:1) and ^H-NMR data (shown below) of this product coincided with those of the isomer F3 described in Reference Exazcple -4. In addition, when this product was checked by HPLC analysis using a chiral column, its HPLC retention time coincided with the retention time of the optical isomer F3 described in reference Example 4. In consequence, it was confirmed that this product (isomer A) is the optical isomer F3 described in Reference Example 4. ^H-NMR (400 MHz, CDCI3) 5: 0.28 - 0.40 (2H, m) , 0.41 - 0.45 (IH, m), 0.53 - 0.63 (IH, m) , 0.72 - 0.81 (IH, m), 1,43 (9H, s), 1.67 - 1.79 (IH, m), 2.03 - 2.09 (IH, m) , 2.37 - 2.40 (IH, m) , 2.95 -3.08 (IH, m), 3.16 (IH, t, J = 10.74 Hz), 3.32 - 3.39 (IH, m) , 3.54 " 3.69 (2H, m) , 4.48 (IH, brs) , 5.13 (2H, s) , 7.31 - 7.37 (5H, m)
HPLC analysis conditions;
Column: CHIRALPAKAD (Daicel Chemical Industries), 0.46 cm x 25 csn Mobile phase: n-hexane:2-propanol = 80:20 (v/v)
Flow rate: 1.0 ml/min
Temperature: room teooperature
Detection: UV (254 nm)
Retention time of isomer A (F3) : 8.16 minutes
Optical pxirity: 99% e.e.
[Reference Example 13] 3- [1-(S)-tert-Butoxycarbonylamino-1-
cyclopropyl]methylpyrroliciine (isomer A: F3)
l-Ben2yl-3- [1- (S) -tert-butoxycaxbonylamino-1-cyclppropyllmethylpyrrolidine (isomer A; 744 mg, 2.25 mmol) was dissolved in dry ethanol (30 ml) , and the solution was mixed with 10% palladium on carbon catalyst (water content 50%; 750 mg) and stirred at 45°C (external temperature) for 1 hour in an atmosphere of hydrogen under atmospheric pressure. The reaction solution was filtered through celite, and the resulting filtrate was concentrated under a reduced pressure, thereby obtaining 542 mg ((quantitative) of the title conpound as white crystals. ^H-NMR (400 MHz, CDCI3) 5: 0.27 - 0.42 (2H, m) , 0.53 - 0.57 (IH, m), 0.74 - 0.81 (IH, m), 1.43 (9H, s), 1.55 - 1.60 (IH, m) , 1.89 - 1.95 (IH, m), 2,23 - 2.26 (3H, m), 2.73 - 2.77 (IH, m) , 2,85 -2,90 (IH, m), 2.95 - 3.01 (2H, m)
[Inventive Example 5] 5-Amino-7"[3-[1-(S)-amino-l-cyclopropyl]methylpyrrolidin-l"yl] -6,8-dif luoro-l- [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline"3-carboxylic acid (the substituent at the 7-position is originated from isomer A, F3)
3- [1- (S) -tert-Butoxycarbonylamino-1-
cycloprppyl]inethylpyrrolidine (isomer A: F3; 541 wg, 2.25 noDol) and triethylamine (6 ml) were added to dry acetonitrile (30 ml) , and the mixture was ftirther mixed with 5-amino-6,7,8-trifluoro-1- [2- (S) -f luoro-1- (R) -cycloprppyl] -1,4-dihydro-4-oxoqainoline-3-carbo^lic acid (548 mg^ 1.73 mmol) and heated under reflux for 15 hours. After cooling of the reaction solution, the thus precipitated crystals were collected by filtration, washed with acetonitrile, mixed with concentrated hydrochloric acid (15 ml) under ice-cooling and then stirred at room temperature for 5 minutes. This was mixed with distilled water (15 ml) , and the thus obtained acidic aqueous solution was washed with dichloromethane (20 ml x 3) , adjusted to pH 11 with sodium hydroxide aqueous solution tmdar ice-cooling and then washed with chloroform (30 ml) . The resulting basic aqueous solution was adjusted to pH 7,4 with 1 N hydrochloric acid and extracted with chloroform (100 ml x 5) . After drying over anhydrous sodixma sulfate, the solvent was evaporated -under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from an ethanol-28% aqueous ammonia and then dried lander a reduced pressure, thereby obtaining 569 mg (75.3%) of the title conpound as yellow needle crystals.
The Rf value of TLC (thin layer chromatography, developed with under layer chloroform:methanol:water = 7:3:1) and ^H-NMR data (shown below) of this product coincided with the data described in Inventive Exanyle 3. ^H-MdR (400 MHz, 0.1 N NaOD) 5: 0.13 - 0.17 (IH, m) , 0.21 - 0.29
(IH, m), 0.41 - 0.48 (IH, m) , 0.51 - 0.61 (IH, m) , 0.73 - 0.82 (IH, m), 1,43 - 1.53 (3H, m) , 1.81 (IH, t, J = 8.79 Hz), 2.10 -2.27 (2H, m), 3.41 - 3.47 (IH, m) , 3.49 - 3.58 (IH, m) , 3.59 -3.66 (IH, m), 3.73 - 3.81 (2H, m) , 4.95 (IH, dm, J = 65.11 Hz), 8.19 (IH, s)
Melting point: 192.5 - 194.5*^0 Elemental analysis data for C21H23F3N4O3 • 0,25H20 Calcd.: C, 57.20; H, 5.37; N, 12.71 Found : C, 57.18; H, 5.39; N, 12.78 Specific rotation: [a]D^° = -146.1° (c 0.32, 0.1 N NaOH) [Inventive Example 6] 7-[3-[1-(S)-Amino-l-
cyclopropyl]iDethylpyrrolidin-l"yl] -6-flTioro-l- [2- (S) -fluoro-l-(R) -cyclopropyl] -1,4-dihydro-8-inetho3cy-4-oxoquinoline"3" carboxylic acid (the sribstituent at the 7"position is originated from isomer A, F3)
4- [1- (S) -tert-But03^carbonylamino-l-cyclqpropyl]2nethylpyrrolidine (isomer A: F3; 240 xng, 1.00 mmol) and triethylamine (0,279 ml, 2.00 mmol) were added to dry dimethyl sulfoxide (1,5 ml) . To this was foarther added 6,7-dif luoro-1- [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-8" methyl-4-oxoquinoline-3-carboxylic acid-BF2 chelate (328 mg, 0.909 nnnol) , and the mixture was stirred at room teoperature for 17 hours and then at 35*^C for 4 hours. After concentration of the reaction solution under a reduced pressure, the resulting residue was mixed with water, and the thus precipitated solid was collected by filtration and washed with water. The thus
(detained solid was suspended in a solution of ethanol: water = 10:1, and the suspension was mixed with triethylamine (2 ml) and heated under reflux for 3 hours. After cooling, the reaction solution was concentrated under a reduced pressure, and the resulting residue was dissolved in chlorofozm (100 ml) . The organic layer was washed with 10% citric acid aqueous solution (50 ml) and then dried over anhydrous sodium sulfate. After filtration, the resulting filtrate was concentrated under a reduced pressure, concentrated hydrochloric acid (5 ml) was added drppwise to the thus obtained residue under ice-cooling and then the mixture was stirred at room tegnperature for 30 minutes. The reaction solution was mixed with 1 N hydrochloric acid (5 ml) , the thus obtained yellow acidic aqueous solution was washed with chloroform (50 ml x 5) and adjusted to pH 12.0 with sodium hydroxide aqueous solution, and then the insoluble material was removed by filtration. The resulting basic aqueous solution was adjusted to pH 7,4 with 1 N hydrochloric acid and extracted with chloroform (100 ml x 4) . After drying over anhydrous sodium sulfate, the solvent was evaporated under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from ethanol and then dried xinder a reduced pressure, thereby obtaining 285 mg (67.3%) of the title con5>ound as yellow crystals.
%-bMR (400 MHz, 0.1 N NaOD) 5: 0.01 - 0.05 (IH, m) , 0.29 - 0,32 (IH, m) , 0.39 - 0.40 (IH, m), 0,64 - 0.66 (IH, m), 1.15 - 1,22 (IH, m) , 1.33 - 1.40 (IH, m), 1.43 - 1.54 (IH, m), 1.73 - 1.77
(IH, m), 1.96 - 1.98 (IH, m) , 2.12 - 2.14 (IH, m) , 3.28 - 3.51 (4H, m), 3.42 (3H, s) , 3.81 - 3.86 (IH, m) , 4.86 (IH, dm, J = 66,6 Hz), 7.49 (IH, d, J = 4.56 Hz), 8.25 (IH, d, J = 3.42 Hz) Melting point: 197.5 - 198.5°C
Elemental analysis data for C22H25F2N3O4 • 0. 5H2O • 0. 5EtOH Calcd. : C, 59.50; H, 6.28; N, 9.03 Foamd : C, 59.50; H, 6.39; N, 8.87 Specific rotation: [a]D^° = -105.5° (c 0.88, 0.1 N NaOH) [Inventive Exaniple 7] 5-Amino"7-[3-[1-(S)-amino-1-cyclqpro|pyl]inethylpyrrolidin"l-yl] -6-fluoro-l- [2- (S) -fluoro-l-(R) -cyclopropyl] -1,4-dihydro"8-iDethyl-4-oxoqiiinoline"3" carboxylic acid (the substituent at the 7-position is originated from isomer A, F3)
4- [1- (S) -tert-Buto3ycarbonylamino-1 -cyclopropyl]inethylpyrrolidine (481 mg, 2.00 mmol) and triethylaznine (1.5 ml) were added to dry dimethyl sulfoxide (2 ml) , and the mixture was further mixed with 5-amino-6,7-dif luoro-1" [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-8-inethyl-4-oxoquinoline-3-carbo3qflic acid (416 mg, 1.33 mmol) and heated under reflux for 72 hours in an atmosphere of nitrogen. After concentration of the reaction solution under a reduced pressure, the resulting residue was dissolved in chloroform (100 ml) , The organic layer was washed with 10% citric acid aqueous solution (50 ml) and then dried over anhydrous sodium sulfate. After filtration, the resulting filtrate was concentrated under a reduced presstire, concentrated hydrochloric acid (5 ml) was
added drqpwise to the thus obtained residue under ice-cooling and then the mixture was stirred at room tenperature for 30 minutes. The reaction solution was mixed with 1 N hydrochloric acid (5 ml) ^ the thus obtained yellow acidic aqueous solution was washed with chloroform (50 ml x 5) and adjusted to pH 12.0 with sodium hydroxide aqueous solution, and then the insoluble material was removed by filtration. The resulting baisic acpieous solution was adjusted to pH 7.4 with 1 N hydrochloric acid and extracted with chloroform (100 ml x 3) . After drying over anhydrous soditmi sulfate, the solvent was evaporated oinder a reduced pressure. Thereafter, the-resTilting residue was purified by a preparative thin-layer chromatography (developed with under layer of chloroform:methanol:water = 7:3:1) , recrystallized from isopropyl alcohol and then dried under a reduced pressure, thereby obtaining 70.0 mg (12.1%) of the title confound as yellow crystals.
^H-NMR (400 MHz, 0,1 N NaOD) 5: 0.01 - 0.10 (2H, m) , 0.27 - 0.39 (2H, m), 0.58 - 0.64 (IH, m) , 0.88 - 0.97 (IH, m) , 1.33 - 1.45 (2H, m), 1.71 - 1.74 (IH, m) , 1.97 - 2.10 (2H, m) , 2,08 (3H, s) , 3.01 - 3,05 (IH, m), 3.17 - 3.21 (IH, m), 3.34 - 3,38 (IH, m) , 3,58 - 3,62 (IH, m), 3.75 - 3.79 (IH, m), 4.90 (IH, dm), 8,14 (IH, s)
Melting point: 226,7 - 227.9°C Elemental analysis data for C22H26^2N403
Calcd.: C, 61.10; H, 6.06; N, 12.96
Fotond : C, 60,84; H, 6.07; N, 12,98
Specific rotation: [a]D^° = -329.0° (c 0.20, 0.1 N NaOH) [Inventive Example 8]
7- [3- [1- (S) -Aini.no-l-cyclopropyljinethylpyrrolidin-l-yl] -1-cyclofpropyl-1,4-dihydro-8"inetho3cy-4"Oxoquinoline"3-carboxylic acid (the si±>stituent at the 7-position is originated from isomer A^ F3)
4- [1- (S) -tert-Butoacycarbonylamino-l-cyclopropyllinethylpyrrolidine (isomer A: F3; 330 mg, 1.37 xnmol) and triethylamine (0.485 ml, 3.48 mmol) were added to 3.0 ml of dry dimethyl sulfoxide, and the mixture was further mixed with 6-fluoro-l-cyclopropyl-1,4-dihydro-8-metho3r7-4-oxoc[uinoline-3-carboxylic acid (321 mg, 1.16 mmol) and stirred at 100°C for 15 hours. After concentration of the reaction solution under a reduced pressure, the resulting residue was dissolved in 100 ml of chloroform. The organic layer was washed with 50 ml of 10% citric acid aqueous solution and then dried over anhydrous sodium sulfate. After filtration, the resulting filtrate was concentrated under a reduced pressure, 5 ml of concentrated hydrochloric acid was added dropwise to the thus obtained residue under ice-cooling and then the mixture was stirred at room temperature for 30 minutes. The reaction solution was mixed with 5 ml of 1 N hydrochloric acid and the thus obtained yellow acidic aqueous solution was washed with chloroform (50 ml X 4) , the resulting insoluble material was removed by filtration, and then the solution was adjusted to pH 12.0 with sodium hydroxide aqueous solution. The resulting basic aqueous
solution was adjusted to pH 7.4 with 1 N hydrochloric acid and extracted with chloroform (100 ml x 4) . After drying over anhydrous sodium sulfate, the solvent was ev^xDrated under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from ethanol-aqueous ammonia and then dried under a reduced pressure, thereby obtaining 230 mg (49.9%) of the title ccaiEpound as yellow crystals. ^H-NMR (400 MHz, 0.1 N NaOD) 5: 0.17 - 0.18 (IH, m) , 0.28 - 0,29 (IH, m) , 0.46 - 0.47 (IH, m), 0,57 - 0.58 (IH, m), 0,77 - 0.79 (2H, m), 1.02 " 1.04 (2H, m), 1.17 - 1.21 (2H, m), 1.69 - 1.81 (IH, m) , 1.92 " 1.95 (IH, m), 2.18 - 2,27 (IH, m), 2.35 - 2,40 (IH, m), 3.33 - 3.52 (3H, m), 3.52 (3H, s), 4.01 - 4,11 (IH, m) , 7,03 (IH, d, J = 8.79 Hz), 7.91 (IH, d, J = 9.03 Hz), 8.48 (IH,
s)
Melting point: 220 - 221°C
Elemental analysis data for C22H27N3O4 ■ 0. 5H2O
Calcd.: C, 65.74; H, 6.90; N, 10,45
Found : C, 65,96; H, 6.90; N, 10.36 [Reference Exaniple 14]
1" [1- (R) -Phenylethyl] -5"Oxopyrrolidine-3" (R) - (N-methyl-N-methoxy) carboxanii de
Oxalyl chloride (6.54 ml, 75.0 mmol) and dimethylformamide (3 drops) were added to dichloromethane solution (200 ml) of 1-[1-(R)-phenylethyl]-5-oxopyrrolidine-3-(R)-carboxylic acid (11.7 g, 50.0 mmol) under ice-cooling, and the mixture was stirred at room ten^^erature for a whole day and
night. After evc^x^ration of the solvent under a reduced pressure, toluene (100 znl) was added to the resulting residue and then the solvent was again evaporated under a reduced pressure. The thus obtained residue was mixed with dichloromethane (200 ml) and N^O-methylhydroxylamine hydrochloride (5.47 g, 55.5 xnmol) , and to the mixture, vdiile stirring under ice-cooling, was then added drppwise dichloromethane solution (50 ml) of triethylamine (17.4 ml, 125 mmol) in 15 minutes. This was stirred under ice-cooling for 30 minutes and then at room temperature for 3 hours. The reaction solution was washed with 10% citric acid aqueous solution (100 ml) , water (100 ml) and saturated soditm bicarbonate aqueous solution (100 ml) in that order and then dried over anhydrous sodium sulfate. The solvent was evaporated xmder a reduced pressure, and the resulting residue was ^>plied to a silica gel colomin chromatography and eluted with a gradient of from chloroform:methanol = 50:1 to 20:1, thereby obtaining 11.3 g (82%) of the title compound as a brown oil.
^H-NMR (400 MHz, CDCl^) 5: 1.54 (3H, d, J = 6.84 Hz), 2.65 (IH, dd, J = 9.77, 7.09 Hz) , 2.77 (IH, dd, J = 8.79, 7.09 Hz) , 3.12 -3.18 (IH, m), 3.20 (3H, s) , 3.37 - 3.48 (IH, m) , 3,55 - 3,64 (IH, m), 3.65 (3H, s) , 5.50 (IH, q, J = 6.84 Hz), 7.28 - 7.37 (5H, m) [Reference Example 15] 4" (R) "Cyclobutylcarbonyl-1- [1- (R) ^henylethyl] -2-pyrrolidone
In an atmosphere of nitrogen, cyclobutylmagnesium chloride (1 N tetrahydrofuran solution, 28 ml) prepared from
chlorocyclobutazie was added drppwise to tetrahydrofuran solu-tion (50 ml) of l"[l-{R)-phenylethyl3-5-oxopyrrolidine"3-(R)-(N-methyl-N-metho3cy) carboxamide (1.93 g, 7,00 mmol) , and the mixture was stirred at room tenperature for 30 minutes. The reaction solution was mixed with 1 N hydrochloric acid (50 ml) under ice-cooling and extracted with ethyl acetate (80 ml x 2) , and the organic layer was washed with saturated sodium chloride aqueous solution (100 ml) and then dried over anhydrous sodium sulfate. The solvent was evaporated under a reduced pressure, and the thus obtained residue was applied to a silica gel colxsmn chrcxnatography and eluted with n-hexane: ethyl acetate — 1:2, thereby obtaining 1.47 g (78%) of the title conpound as a light yellow oil.
^H-NMR (400 MHz, CDCI3) 5: 1.53 (3H, d, J = 7.33 Hz), 1.78 - 1.89 (IH, m) , 1.92 - 2.06 (IH, m) , 2,06 - 2.31 (4H, m) , 2,58 - 2.65 (2H, m) , 3.05 (IH, dd, J = 9.28, 8,79 Hz), 3.13 - 3.21 (IH, m) , 3.31 (IH, quint, J= 8.30 Hz), 3.53 (IH, dd, J = 9.28, 6.83 Hz), 5.48 (IH, q, J = 7.33 Hz), 7.27 - 7,37 (5H, m) [Reference Exanple 16]
4" (R) - (l-Cyclobutyl-l-hydroxy)methyl-l- [1- (R) -phenylethyl] -2-pyrrolidone
Under ice-cooling, sodium borohydride (295 mg) was added to ethanol (40 ml) solution of 4-(R)-cyclobutylcarbonyl-l"[1" (R)-phenylethyl]-2-pyrrolidone (2.12 g, 7.80 mmol) , and the mixture was stirred at the same temperature for 1 hooir. The reaction solution was mixed with 10% citric acid (50 ml) \inder
ice-cooling, ethanol was ev^x^rated under a reduced pressiire^ the thus obtained residue was extracted with chlorofonn (80 ml x 2) f and then the organic layer was washed with saturated sodium chloride acpieous solution (100 zol) and dried over anhydrous sodium sulfate. The solvent was evc^x^rated under a reduced pressure, and the thus obtained residue was applied to a silica gel column chromatography and eluted with a gradient of from n-hexane:ethyl acetate = 1:3 to ethyl acetate, thereby obtaining 2.10 g (98%) of the title ccxnpound as a light yellow oil (isomer mixture).
^H-NMR (400 MHz, CDCla) 5: 1.50 (3H, d, J = 6.83 Hz), 1.68 - 2.01 (6H, m), 2.14 " 2.45 (3H, m) , 2.45 - 2.56 (IH, m) , 2.91 - 3.05 (IH, m), 3.19 - 3.31 (IH, m) , 3.41 - 3.49 (IH, m) , 5.42 - 5.49 (IH, m), 7,24 - 7.36 (5H, m) [Reference Exanple 17]
4- (R) - (l-A2ido--l-cyclobutyl)methyl-l- [1- (R) -phenylethyl] -2-pyrrolidone
Under ice-cooling, triethylamine (1.36 ml, 9.80 mmol) and then methanesulfonyl chloride (640 |il, 8.30 mmol) were added to dichloromethane (35 ml) solution of 4-(R)-(l-cyclobutyl-l-hydroxy)methyl-1- [1- (R) -phenylethyl] -2--pyrrolidone (2.05 g, 7.50 mmol) , and the mixture was stirred at the same tesiperature for 1 hour. The reaction solution was mixed with 10% citric acid (35 ml) under ice-cooling and extracted with chloroform (50 ml x 2) , and the organic layer was washed with saturated sodium chloride aqueous solution (150 ml) and then dried over anhydrous sodium
sulfate. After evaporation of the solvent under a reduced pressure, the thus obtained residue was dissolved in N,N'-diTnethylformamide (30 ml) , cuid the solution was mixed with sodium azide (1.46 g, 22.5 mmol) and stirred at 60^C for 3 hours. After cooling, the reaction solution was mixed with water (150 ml) under ice-cooling and extracted with ethyl acetate (150 ml x 3) , and the organic layer was washed with saturated sodium chloride aqueous solution (150 ml) and then dried over anhydrous sodium sulfate. After evaporation of the solvent under a reduced pressure, the thus obtained residue was ^plied to a silica gel column chromatogr^hy and eluted with n-hexane: ethyl acetate = 3:2, thereby obtaining 898 mg (40%) of a low polarity title coiopound (isomer Bl) as a colorless oil and then with n-hexane:ethyl acetate = 2:3, thereby obtaining 847 mg (38%) of a high polarity title coopound (isomer B2) as colorless crystals. Isomer Bl
^H-NMR (400 MHz, CDCI3) 5: 1.52 (3H, d, J = 6.83 Hz), 1.72 - 2.01 (5H, m), 2.07 - 2.17 (IH, m) , 2,26 - 2.41 (3H, m) , 2.45 - 2.56 (IH, m) , 2.98 (IH, dd, J = 9.77, 7.81 Hz), 3.14 (IE, dd, J = 9.77, 7.32 Hz), 3.32 (IH, dd, J = 8.76, 3.91 Hz), 5.47 (IH, q, J = 6.83 Hz), 7,25 - 7,35 (5H, m) Isomer B2
^H-NMR (400 MHz, CDCI3) 5: 1.52 (3H, d, J = 6.83 Hz), 1.75 - 2.03 (5H, m) , 2,03 - 2.17 (IH, m) , 2.19 - 2.38 (2H, m) , 2.40 - 2.56 (2H, m) , 2.99 (IH, dd, J = 9.77, 8,30 Hz), 3.14 (IH, dd, J = 9.77, 7.32 Hz), 3,30 (IH, dd, J = 8.30, 6.34 Hz), 5.47 (IH, q, J
= 6.83 Hz), 7.25 - 7.35 (5H, m)
[Reference Example 18]
4- (R) - [1- (tert-Butoxycazbonyl) aininO"l"cyclobutyl]inethyl-l- [1-
(R) -phenylethyl] -2-pyrrolidone (iscaner Bl)
Ethanol (50 ml) solution of 4-(R) - (1-azido-l-
cyclobutyl)loethyl-l- [1- (R) -phenylethyl] -2-pyrrolidone (isomer Bl) (835 tag, 2.80 xrsnol) was mixed with 10% palladium on carbon catalyst (water content 53.8%, 850 mg) , and 5 hours of catalytic hydrogenation was carried out at room temperature in an atmosphere of hydrogen under atmospheric pressure. The reaction solution was filtered and the solvent was evaporated under a reduced pressure. The resulting residue was dissolved in dichloromethane (20 ml) ^ and the solution was mixed with di-tert-butyl bicarbonate (917 mg) and triethylamine (780 |il) and stirred at room temperature for 15 hours. The reaction solution was mixed with chloroform (50 ml) and washed with 10% citric acid (80 ml) and water (80 ml) and then the organic layer was dried over anhydrous sodium sulfate. The solvent was esvaporat/ed under a reduced pressure, and the thus obtained residue was applied to a silica gel colxmin chromatogr^hy and eluted with a gradient of n-hexane:ethyl acetate = 3:2 to 1:1, thereby obtaining 809 mg (78%) of the title cQa^>ound as white amorphous. ^H*NMR (400 MHz, CDCI3) 5: 1.44 (9H, s) , 1.48 (3H, d, J = 7,32 Hz), 1.66 - 1.98 (6H, m) , 2.17 - 2.43 (4H, m) , 2.94 - 3,03 (IH, m) , 3-09 - 3,18 (IH, m) , 3,59 - 3.68 (IH, m) , 4.46 - 4,58 (IH, m) , 5,46 (IH, q, J = 7.32 Hz), 7.27 - 7,35 (5H, m)
[Reference Exaicple 19]
4" (R) - [1- (tert-Butoxycarbonyl) amino-l-cycldbutyllioethyl-l- [1-
(R) -phenylethyl] -2-pyrrolidoiie (isomer B2)
Ethanol (40 ml) solution of 4-(R) -(l-azido-l-cyclobu-b^l) methyl-l- [1- (R) -phenylethyl] -2-pyrrolidone (isomer B2) (776 ing, 2.60 nmaol) was mixed with 10% palladitsm on carbon catalyst (water content 53,8%, 800 mg) and 5 hours of catalytic hydrogenation was carried out at room tpmperature in an atciosphere of hydrogen Tuxder atmospheric pressure. The reaction solution was filtered and the solvent was evaporated under a reduced pressure. The thus obtained residue was dissolved in dichloromethane (20 ml) , and the solution was mixed with di-tert-butyl bicarbonate (851 mg) and triethylamine (725 pi) and stirred at room temperature for 15 hours. The reaction solution was mixed with chloroform (50 ml) and washed with 10% citric acid (80 ml) and water (80 ml) and then the organic layer was dried over anhydrous sodium sulfate. The solvent was evaporated under a reduced pressure^ and the thus obtained residue was applied to a silica gel column chromatography and eluted with a gradient of n-hexane:ethyl acetate = 1:1 to 2:3, ther^^y obtaining 846 mg (87%) of the title compotind as white amorphous. ^H-NMR (400 MHz, CDCI3) 5: 1.43 (9H, s) , 1.50 (3H, d, J = 7,32 Hz), 1.70 - 1.96 (6H, m) , 2,08 - 2.22 (IH, m) , 2,22 - 2.36 (2H, m) , 2,36 - 2.47 (IH, m) , 2.96 (IH, dd, J = 9.27, 8.30 Hz), 3.10 (IH, dd, J = 9,27, 8.79 Hz), 3.55 - 3.62 (IH, m) , 4.28 (IH, d, J = 9.77 Hz), 5,46 (IH, q, J = 7.32 Hz), 7.25 - 7.35 (5H, m)
[Reference Exanple 20]
3- (R) - [1- (tert-Biitoxyca2±)onyl) ami no-l-cyclobutyllinethyl-l- [1-
(R)-phenylethyl] pyrrolidine (isomer Bl)
In an atmosphere of nitrogen, 1 M borane-tetrahydrofuran
cociplex solution (5.6 ml) was added drppwise to tetrahydrofuran
solution (15 ml) of 4-(R) - [1-(tert-butoxycarbonyl) amino-1-cyclobu"^l]methyl-l- [1- (R) -phenylethyl] -2-pyrrolidone (isomer Bl) (700 mg, 1.88 mmol) under ice-cooling, and the mixture was stirred at room tenqperatnire for 13 hoxirs. The solvent was evaporated under a reduced pressure, and the resul-bing residue was mixed with 80% aqueous ethanol (15 ml) and triethylamine (3 ml) and heated under refliox for 4 hotirs. After cooling, the solvent was evaporated under a reduced pressure, and the thus obtained residue was mixed with chloroform (30 ml) , washed with water (10 ml) and saturated sodium chloride aqueous solution (10 ml) and then dried over anhydrous sodium sulfate. The solvent was evaporated xmder a reduced pressiure, and the thus obtained residue was applied to a silica ^1 column chromatography and eluted with chloroform:methanol = 20:1, thereby obtaining 565 mg (84%) of the title conpound as colorless crystals. ^H-NMR (400 MHz, CDCI3) 5: 1.36 (3H, d, J = 6.8
(R) -phenylethyl ] pyrrolidine (isomer B2)
In an atznosphere of nitrogen, 1 M borane-tetrahydroforan conplex solution (6.4 ml) was added dropwise to tetrahydrofuran solution (15 ml) of 4-(R)-[1-(tert-butoxycarbonyl)amino-l-cyclobutyl]methyl-l- [1- (R) -phenylethyl] -2-pyrrolidone (isomer B2) (797 xng, 2.14 mmol) tinder ice-cooling, and the mixture was stirred at room temperature for 13 hours. The solvent was evaporated under a reduced pressure, and the resulting residue was mixed with 80% aqueous ethanol (15 ml) and triethylamine (3 :ml) and heated under reflux for 4 hours. After cooling, the solvent was evaporated under a reduced pressure, and the thus obtained residue was mixed with chloroform (30 ml) , washed with water (10 ml) and saturated sodium chloride aqueous solution (10 ml) and then dried over anhydrous sodium siilfate. The solvent was evaporated xmder a reduced pressure, and the thus obtained residue was applied to a silica gel column chromatography and eluted with chloroform:methanol = 20:1, thereby obtaining 743 mg (97%) of the title cavapovind as colorless oil.
^H-NMR (400 MHz, CDCI3) 5: 1.37 (3H, d, J = 6,83 Hz), 1.46 (9H, s) , 1.64 - 1.93 (8H, m), 2.10 - 2.30 (3H, m) , 2,30 - 2,51 (IH, m) , 2.51 - 2.69 (2H, m) , 3.11 - 3,23 (IH, m) , 3,43 - 3.52 (IH, m) , 4.92 - 5.01 (IH, m), 7.22 - 7.32 (5H, m) [Reference Example 22]
3"(R)-[1-(tert-Butoxynarbonyl) amino-1-cyclcbutyl]methylpyrrolidine (isomer Bl)
10% Palladium on carbon catalyst (water content 53,8%,
500 mg) was added to ethanol (30 ml) solution of 3-(R) - [1-(tert-
butoxycarbonyl) amino-l-cyclobutyl]inethyl-l- [1- (R) -phenylethyl]pyrrolidine (isomer Bl) (516 mg^ 1.44 mmol) , and the mixture was sx±)jected to 5 hoTirs of catalytic hydrogenation at an external temperature of 50°C in an atmosphere of hydrogen under atmospheric pressure. The reaction solution was filtered and the solvent was evaporated under a reduced pressure^ thereby obtaining 366 mg (quantitative) of the title conpoimd as colorless crystals. This product was used in the subsequent reaction without purification. [Reference Example 23]
3"(R)"[1-(tert-Butoxycarbonyl) amino-1-cyclobutyl]methylpyrrolidine (isccaer B2)
10% Palladium on carbon catalyst (water content 53.8%, 650 mg) was added to ethanol (40 ml) solution of 3-(R)-[1-(tert-butoxycarbonyl) amino-l-cyclobutyl]methyl-l- [1- (R) -phenylethyl]pyrrolidine (isonier B2) (645 mg, 1.80 mmol) , and the mixture was subjected to 5 hours of catalytic hydrogenation at an external tenperature of 50°C in an atmosphere of hydrogen lander atmospheric pressure. The reaction solution was filtered and the solvent was evaporated under a reduced pressure, thereby obtaining 458 mg (quantitative) of the title cooopound as colorless crystals. This product was used in the subsequent reaction without purification. [Inventive Example 9] 5-Amino"7- [3- (R) - (l-amino-l-cyclobutyl) methyl] pyrrolidin-1-yl] -
6 r 8-dif luoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4"dihydro-4-oxoc[uinoline-3-carboxylic acid (the snbstitaient at the 7-position is originated frcan Bl)
5-Amino-6,7,8-trifluoro-1-[2-(S)-fluoro-1-(R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid (379 mg, 1.20 mmol), 3-(R)-[1-(tert-butoxycarbonyl)amino-1-cyclabutyllmethylpyrrolidine (isomer Bl) (366 xog, 1.44 mmol) and triethylamine (3 ml) were added to acetonitrile (15 ml) and heated under reflux for 8 hours. After cooling, the reaction solution was concentrated under a reduced pressure, and the thus obtained residue was mixed with concentrated hydrochloric acid (15 ml) under ice-cooling and then stirred at room tesqperature for 10 minutes. This hydrochloric acid solution was washed with chloroform (20 ml x 3) , and was made alkaline by adding 30% sodiimi hydroxide aqueous solution under ice-cooling and then stirred at rocm temperature for 1 hour. This suspension was adjusted to pH 7.6 by adding concentrated hydrochloric acid and 1 N hydrochloric acid and then extracted with chloroform (100 ml X 3) , The organic layer was dried over anhydrous sodium sulfate and the solvent was evaporated under a reduced pressure, trhereafter, the resulting residue was purified by recrystallizJLng from ethanol-n-hexane and then dried \ander a reduced pressure, thereby obtaining 386 mg (74%) of the title compound as light yellow crystals.
^H-MMR (400 MHz, 0.1 N NaOD) 5: 1.22 - 1.83 (IIH, m) , 1.83 -1.97 (IH, m) , 1.97 - 2.18 (2H, m) , 2.18 - 2.29 (IH, m) , 3.07 -
3.28 (2H, m), 3.35 - 3.46 (IH, m) , 3.53 - 3.69 (2H, m) , 4.78 -
4.89 (0.5H, m), 4.93 - 5.02 (0.5H, m) , 8.17 (IH, s) Melting point: 175.3 - 177,6°C (deconp.)
Elemental analysis data for C22H25P3N4O3
Calcd.: C, 58.66; H, 5.59; N, 12.44 Fotind : C, 58.55; H, 5.61; N, 12.33
[Inventive Example 10]
5-Amino"7- [3- (R) ■ (l-amino-l-cyclobutyl) methyl Jpyrrolidin-l-yl] -
6,8-dif liioro-1" [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline--3-carbo3cylic acid (the soabstituent at the 7-position is originated from B2)
5-Amino-6,7,8-trif luoro-l- [2- (S) -f luoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carbo3cylic acid (474 mg, 1.50 mmol) , 3- (R) - [1- (tert-butoarycarbonyl) amino-l-cyclQbutyl]inethylpyrrolidine (isomer B2) (458 xng, 1.80 mmol) and trie thy lazaine (4 ml) were added to acetonitrile (20 ml) and heated under reflux for 8 hours. After cooling, the reaction solution was concentrated under a reduced pressure, and the thus obtained residue was mixed with concentrated hydrochloric acid (15 ml) under ice-cooling and then stirred at room temperature for 10 minutes. This hydrochloric acid solution was washed with chloroform (20 ml x 3) , and was made aUcaline by adding 30% sodium hydroxide aqueous solution tmder ice-cooling and then stirred at room temperature for 1 hour. This suspension was adjusted to pH 7.6 by adding concentrated hydrochloric acid and 1 N hydrochloric acid and then extracted with chloroform (100 ml
X 3) . The organic layer was dried over anhydrous soditnn stilfate and the solvent: was ev^x^rated under a reduced pressure. Thereafter, the resulting residue was purified by recrystallizing from chloroform-n-hexane and then dried tinder a reduced pressure, thereby obtaining 386 mg (74%) of the title com^xDund as light yellow crystals.
^H"NMft (400 MHz, 0.1 N NaOD) 5: 1.27 - 1.76 (7H, m) , 1.76 - 1.99 (6H, m), 2.17 " 2.28 (IH, m) , 2.34 - 2.42 (IH, m) , 3.36 - 3.50 (2H, m) , 3.50 - 3.59 (IH, m) , 3.64 - 3.79 (2H, m) , 4.79 - 4.89 (0.5H, m) , 4.98 - 5.06 (0.5H, m), 8.19 (IH, s) Melting point: 198.2 - 201.4°C (decamp.) Eleanental analysis data for C22H25F3N4O3 • 1. OH2O Calcd.: C, 56.40; H, 5.81; N, 11,96 Found : C, 56.34; H, 5.84; N, 11,75 The antibacterial activity of each conpound of the present invention was measured in accordance with the standard method specified by the Japan Society of Chemotherapy, with the results shown in the following Table 2 as MIC vauLues (^g/ml) . In this connection, MIC values of levofloxacin (LVEX) and ciprofloxacin (CPFX) are also shown for the sake of conparison with the MIC values of the ccanpounds of the present invention.
INDUSTRIAL APPLICABILITY
The compound of the present invention is possessed of excellent antibacterial action against a broad range of Gram-negative and Gram-positive bacteria, particularly showing strong antibacterial activity against methicillin-resistant Staphylococcus axureus, penicillin-resistant pneumococcus, enterocuccus and the like Gram-positive bacteria and quinolone-resistant bacteria, and is also possessed of excellent safety and good pharmacokinetics, such as attenuation of micronucleax test, so that it is useful as an antibacterial compound to be used in the chonotherapy of bacterial infections.
CLAIMS 1, A ccttpound represented by the following formula (I), its salts and hydrates thereof:
wherein R1 and R2 each independently represents a hydrogen atom or an alJcyl group having 1 to 6 carbon atoms, in which the alkyl groxjp may have one or more substituents selected frccn the group consisting of a hydroxyl group, a halogen atom, an alkylthio group having 1 to 6 carbon atoms and an alkyloxy group; n is an integer of 1 to 4; and
Q is a partial structure represented by the following formula (la) :
wherein R^ represents an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, a halogenoalkyl group having 1 to 6 carbon atoms, a cyclic alkyl
group having 3 to 6 carbon atoms vrtiich may have a substituent, an axyl grox^J which niay have a sxabstituent, a heteroaryl group vdiich niay have a substituent, an alkoaofl groi^ having 1 to 6 carbon atoms or an alkylamino groxjp having 1 to 6 carbon atoms/
R^ represents a hydrogen atom or an alkylthio groi^) having 1 to 6 carbon atoms;
R^ and R^ may form together with a part of the mother skeleton a ring structure optionally containing a sulfur atom as a ring constituting atom thereof and optionally having an alkyl groxxp having 1 to 6 carbon atoms as a substituent;
P? represents a hydrogen atom, an amino grox:^?, a hydroxyl group, a thiol groi^j, a halogenomethyl group, an alkyl groijp having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl groxap having 2 to 6 carbon atoms or an alkoxyl groixp having 1 to 6 carbon atoms, in which the amino group may have one or more substituents selected from the groo^ consisting of a foniYl group, an alkyl group having 1 to 6 carbon atoms and an acyl group having 2 to 5 carbon atoms;
X^ represents a halogen atom or a hydrogen atom; A*" represents a nitrogen atom or a partial structure represented by formula (II) :
vrtierein X^ represents a hydrogen atom, an amino group^ a halogen atom, a cyano group, a halogenomethyl group, a
halogenonethdxyl group, an alJcyl group having 1 tx> 6 carbon atxxns, an alkenyl grox^^ having 2 to 6 carbon atoms, an allcynyl group having 2 to 6 carbon atcins or an alkoxyl group having 1 to 6 carbon atoms, in which the amino group may have one or more substituents selected from the groiip consisting of a formyl groi^, an alkyl group having 1 to 6 carbon atoms and an acyl group having 2 to 5 carbon atoms; and
X^ and R^ may form together with a part of the mother skeleton a ring structure optionally containing an oxygen atom, a nitrogen atom or a sulfur atom as a ring constituting atom thereof and optionally having an alkyl group having 1 to 6 carbon atoms as a substituent; and
Y represents a hydrogen atom, a phenyl group, an acetoxymethyl group, a pivalcyloxymethyl groa:p, an ethccKycarbonyl group, a choline group, a dimethylaminoethyl groiq?, a 5-indanyl group, a phthalidinyl grotrp, a 5-alkyl-2-oxo-l,3-dioxol-4-ylmethyl group, a 3-acetoxy-2-oxobutyl groiip, an alkyl group having 1 to 6 carbon atoms, an alkoxymethyl group having 2 to 7 carbon atoms or a phenylalkyl groip conposed of an alkylene groxjp having 1 to 6 carbon atoms and a phenyl grox:p.
2, A conpound, its salts and hydrates thereof according to
claim 1, vdierein Q in the formula (I) is a S-carfaoay-g-fluoro-
2,3-dihYdro-3" (S) -methyl-7-oxo-7H-pyrido [1,2,3-de] [1.4]benzoxazin-10-yl group,
3. A conpound, its salts and hydrates thereof according to
claim 1 or 2, lAerein the conpound of formula (I) is a
stereochemically pure conpound,
4. A accqpoundf its salts and hydi^tes thereof according to claim 1, wherein R^ in the formula (I) is a halogenocycloprcpYl group.
5. A conpound, its salts and hydrates thereof according to claim 1, 3 or 4, wherein the halogenocyclopropyl group in the fonnula (I) is a 1,2-cis-halogenocyclqpropyl groa^.
6. A cccrpound/ its salts and hydrates thereof according to claim 5, wherein the halogenocyclopropyl groi:^^ in the foxroula (I) is a stereoch^nically pure substituent.
7. A coirpoimd, its salts and hydrates thereof according to claim 6, wherein the halogenocyclopropyl groijp in the formula (I) is a {1R,2S) "2-halogenocyclopropyl group,
8. A conpound, its salts and hydrates thereof according to claim 7, wherein the halogen atcm of the halogenocyclopropyl group in the fonnula (I) is a fluorine atom.
9. A conpound, its salts and hydrates thereof according to claim 8, wherein the ccnpound of formula (I) is a stereochemically pure ccnpound.
10. A cocrpound, its salts and hydrates thereof according to claim 1, 2, 3, 4, 5, 6, 1, 8 or 9, wherein n in the formula (I) is 1.
11. A ccnpound, its salts and hydrates thereof according to claim 10, wherein the conpoiond of formxila (I) is a stereochenically pure ccnpound.
12. 7- [3- [1- (S) -Amino-l"-cyclopropyl]methylpyrrolidin-l-
yl]-6"f luoro-1"[2-(S)-f luoro-1-(R)-Gyclaprppyl]-1,4-dihydro-8-inethoxy-4"Oxcx5uinoline-3-carboxylic acid, its salts and hydrates thereof.
13. 5-Amino-7-[3-[l-(S)"ainino-l-
cyclopropyl]inethylpyrrolidin-l-yl] -6-fluoro-l-- [2- (S) -fluoro-1-
(R) -cyclopropyl ] -1,4-dihydro-8"inethyl-4*oxoquinoline-3-carboxylic
aci.d, its salts and hydi^tes thereof.
14. 5-Amino-7-[3-[l-(S)"ainino-l"
cYclopropyl Imethylpyrrolidin-l-yl ] -6,8-difluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro"4-oxoquinoline-3-carboxylic acid, its salts and hydrates thereof.
15. A ccxrpound, its salts and hydrates thereof according to claim 1, 3, 4, 5, 6, 7, 8, 9 or 11, \dierein Y is a hydrogen atom.
16. A drug containing the ccnpoiand, its salts and hydrates thereof described in any one of claims 1 to 15 as an active ingredient.
17. An antibacterial containing the conpound, its salts and hydrates thereof described in any one of claims 1 to 15 as an active ingredient.
18. A compound substantially as herein described and exemplified.
19. An antibacterial containing the compound substantially as herein described and exemplified.
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