| Title of Invention | A METHOD OF SYNTHESIS OF PROTECTED PYRROLOBENZODIAZEPINE COMPOUND OF FORMULA (I) |
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| Abstract | METHOD OF SYNTHESIS OF PROTECTED PYRROLOBENZODIAZEPINES A method of synthesis of a N-10 protected PBD compound of formula (I) via an intermediate of formula (II) or formula (V). |
| Full Text | The present application relates to method of synthesis of protected pyrrolobenzodiazepine (PBD) compounds, and in particular, PBDs having a N-10 protecting group, as well as intermediates in these methods. Background A large number of both synthetic and naturally occurring low molecular weight ligands are known that interact with DNA via a number of different mechanisms, including covalent or non-covalent interaction in the minor or major grooves, intercalation between base pairs or other types of nonspecific interactions. A particular class of compounds which interacts with the minor groove are the pyrrolobenzodiazepines (PBDs). PBDs have the ability to recognise and bond to specific sequence of DNA; the most preferred sequence is PuGPu (Purine-Guanine- Purine ) . The first PBD antitumour antibiotic, anthramycin, was discovered in 1965 (Leimgruber, et al., J. Am. Chem. Soc., 87, 5793-5795 (1965); Leimgruber, et al., J. Am. Chem. Soc, 87, 5791-5793 (1965)). Since then, a number of naturally occurring PBDs have been reported, and over 10 synthetic routes have been developed to a variety of analogues (Thurston, et al. , Chem. Rev. 1994, 433-465 (1994)). Family members include abbeymycin (Hochlowski et al., 1987 J". Antibiotics, 40, 145-148), chicamycin (Konishi et al., 1984 J. Antibiotics, 37, 200-206), DC-81 (Japanese Patent 58-10 487; Thurston et al., 1990, Chem. Brit., 26, 767-772; Bose et al., 1992 Tetrahedron, 48, 751-758), mazethramycin (Kunimoto et al., 1980 J. Antibiotics, 33, 665-667), neothramycins A and B(Takeuchi et al., 1976 J. Antibiotics, 29, 93-96), porothramycin (Tsunakawa et al., 1988 J". Antibiotics, 41, 1366-1373), prothracarcin (Shimizu et al., 1982 J. Antibiotics, 35, 972-978; Langley and Thurston, 1987 J". Org. Chem., 52, 91-97), sibanomicin (DC-102) (Hara et al., 1988 J. Antibiotics, 41, 702-704; Itoh et al., 1988 J. Antibiotics, 41, 1281-1284), sibiromycin (Leber et al., 1988 J. Am. Chero. Soc., 110, 2992-2993) and tomamycin (Arima et al., 1972 J. Antibiotics, 25, 437-444). PBDs are of the general structure: (Figure Removed)They differ in the number, type and position of substituents, in both their aromatic A rings and pyrrolo C rings, and in the degree of saturation of the C ring. There is either an imine (N=C), carbinolamine (NH-CH(OH))or a carbinolamine methyl ether (NH-CH(OMe))at the N10-C11 position which is the electrophilic centre responsible for alkylating DNA. These forms may exist in equilibrium in solution. All of the known natural products have an (S)-configuration at the chiral Clla position which provides them with a right-handed twist when viewed from the C ring towards the A ring. This gives them the appropriate three-dimensional shape for isohelicity with the minor groove of B-form DNA, leading to a snug fit at the binding site (Kohn, 1975 In Antibiotics III. Springer-Verlag, New York, pp. 3-11; Hurley and Needham-VanDevanter, 1986 Ace. Chem. Res., 19, 230-237). Their ability to form an adduct in the minor groove enables them to interfere with DNA processing, hence their use as antitumour agents. The present inventors have previously disclosed that PBDs can be employed as prodrugs by protecting them at the N10 position with a nitrogen protecting group which is removable in vivo (WO 00/12507). Many of these protecting groups are carbamates, and are, for example, of the structure(Figure Removed)where * indicates the attachment point to the N10 atom of the PBD. These protecting groups are described as being added to the compound at two different stages in the synthesis route. One stage is addition of the corresponding chloroformate to a precursor structure as follows, which precursor is then cyclised to form the desired final compound: (Figure Removed)An alternative route discussed involves adding the corresponding chloroformate to a carboxy aniline precursor of the starting material in the above route. These routes employ a chloroformate reacting with an (aromatic) amine to form the carbamate. The present inventors have discovered that the use of the chloroformate has disadvantages, and have therefore developed an alternative synthesis route, which does not employ a chloroformate. Summary of the Invention Accordingly, a first aspect of the present invention provides a method of synthesis of a compound of formula (I): (Figure Removed)comprising the step of either: (a) reacting a compound of formula (II) with a compound of formula (III) to yield a compound of formula (IV): (Figure Removed) (b) reacting a compound of formula (V) with a compound of formula (III) to yield a compound of formula (VI): (Figure Removed) wherein the dotted lines indicate the optional presence of a double bond between Cl and C2 or C2 and C3; R2 and R3 are independently selected from -H, -OH, =0, =CH2, -CN, -R, OR, =CH-R, 0-S02-R, C02R and COR; R6, R7 and R9 are independently H, R, OH, OR, SH, SR, NH2, NHR, NRR', nitro, Me3Sn and halo; R8 is either selected from H, R, OH, OR, SH, SR, NH2r NHR, NRR', nitro, MeaSn and halo or the compound is a dimer with each monomer being the same of different and being of the relevant formula, where the R8 groups of each monomer form together a bridge having the formula -X-R"-X~, where R" is a C3_12 alkylene group, which chain may be interrupted by one or more heteroatoms, e.g. 0, S, NH, and/or aromatic rings, e.g. benzene or pyridine, and each X is independently selected from 0, S and NH; R10 is such that R10-OC(=0)- forms a nitrogen protecting group; R11 is either H or R; R12 is an optionally substituted C1-4 alkyl group; P and Q are such that -CPQ is a masked aldehyde group; wherein R and R' are independently selected from optionally substituted C1-30 alkyl, C3-20 heterocyclyl, and C5-20 aryl. .Reaction conditions The reactions above should be carried in an anhydrous and non-hydroxylic organic solvent, which is preferably non-polar. Suitable solvents include anhydrous dichloromethane and anhydrous toluene. The reaction should be carried out in the presence of a base is present, and suitable bases include pyridine or TEA. The reaction may be carried out at 0°C, or at a higher temperature to increase the rate of reaction. Further .Reaction Steps If the method of synthesis of the first aspect of the invention comprises step (a), then the synthesis of the compound of formula (I) from the compound of formula (IV) will be according to the following scheme: (Figure Removed) wherein T is O or S, and each Rac is independently selected from C3-10 alkyl or together can be a C1-3 alkylene group. Thus the synthesis further comprise the following steps: (i) hydrolysis of the ester -C(=O)OR12; (ii)coupling of resulting acid with a compound of formula: (Formula Removed) TRac wherein M is either OH or (a) when M is OH, oxidation of the resulting compound to yield a compound of formula (Ia); (b) when M is palladium mediated unmasking of the resulting compound to yield a compound of formula (Ia); and (iii) converting the compound of formula (Ia) to a compound of formula (I)by: (a) direct etherification of compound of formula (Ia) or treatment of compound of formula (Ia) with R11SH and preferably Lewis Acid or treatment of compound of formula (Ia) with R11NH and preferably Lewis Acid, depending upon whether X is 0, S or NH, respectively. The ester hydrolysis is usually carried out under mild conditions, for example at 0°C with lithium hydroxide or under non-basic conditions, if the carbamate is sensitive to these. In this situation suitable R12 groups would include allyl, butyl and benzyl. The coupling reaction is preferably carried out under mild conditions, with preferred conditions being DCC or HOBt in DCM at low temperature. Oxalyl chloride and thionyl chloride may be used, but are less preferred. Oxidation of the deprotected primary alcohols provokes spontaneous PBD B-ring closure. A number of oxidizing agents/conditions can be employed to achieve ring closure including; Swern Oxidation, SO3-Pyridine/DMSO, pyridinium dichromate, TPAP/NMO, Dess-Martin Periodinane and TEMPO-DAIB. The TEMPO/DAIB system is particularly favored as it does not require rigorous anhydrous conditions, reaction is easily monitored by TLC, and there is no evidence of over-oxidation to the PBD dilactam species. The palladium mediated demasking can be carried out under literature conditions, such as using bisacetonitrile palladium chloride, (CH3CN)2PdCl2, in acetone (Lipshutz, B.H., et al., Tetrahedron Letters, 26, 705 (1985)). The compound of formula (II) can be synthesised according to the following scheme: (Figure Removed)Thus the synthesis further comprise the following steps: (i) esterification, by reaction with R12OH; and (ii) reaction with triphosgene to form isocynate. The esterification is carried out under normal conditions, the ester is commercially available itself. Often The conversion to the isocyanate can be carried out by the action of phosgene, trichloromethyl chloroformate or triphosgene, of which triphosgene is the preferred agent as is is an easy to handle crystalline solid rather than a toxic gas. The reaction should be carried in an anhydrous and non-hydroxylic organic solvent, which is preferably non-polar. Suitable solvents include anhydrous dichloromethane and anhydrous toluene. The reaction may be carried out at room temperature, and is conveniently monitored by infrared spectroscopy at about 2260 cnf If the method of synthesis of the first aspect of the invention comprises step (b), then the synthesis of the compound of formula (I) from the compound of formula (VI) will be according to the following scheme(Figure Removed)Thus the synthesis comprises the following steps: (i) (a) if -CPQ represents a protected alcohol group, deprotection followed by oxidation; or (b) if -CPQ represents an acetal or thioacetal, palladium mediated unmasking; and (ii) (a) if XR11 is OR11, direct etherification; (b) if XR11 is SR", treatment with RnSH and a catalyst, such as a Lewis Acid, e.g. A12C>3) (c) if XR11 is NHR11, treatment with RnNH and a catalyst, such as a Lewis Acid, e.g. A1203) . The hydroxyl protecting groups (if present) must be removed to allow B-ring cyclization to take place. Removal is under standard conditions. For example, an acetate protecting group can be removed under extremely mild conditions with potassium carbonate. A silyl ether protecting group can be removed by fluoridolysis using TBAF or with mild acid. If these conditions are unsuitable for a particular carbamate, alternative hydroxyl protecting groups can be selected as long as they are capable of surviving the reduction of the nitro group. The conditions for the remaining reactions are as described above in relation to the first method. (Figure Removed)coupling of acid with hydroxymethyl pyrrole; or (b) coupling of acid with acetalmethyl pyrrole; (ii) reduction of aromatic nitro group to form aromatic ainine group; and (ii) reaction with triphosgene to form isocynate. Commercially available nitrobenzoic acids are converted to acid chlorides and coupled to pyrrolidinemthanol and its derivatives under literature conditions. Free hydroxyl groups may be protected as silyl ethers or acetates (other protectings groups, including acetals such as MEM or MOM, can be employed as long as they are stable to the conditions requires to reduced the aromatic nitro group) in order to prevent the formation of a bridging carbamate in the isocyanate formation step. The reduction of the nitro group can be carried out under standard conditions. Preferred methods include hydrogenation over a palladium on charcoal catalyst in a Parr hydrogenator, and using sodium dithionite, tin(II) chloride or Raney Nickel and hydrazine, depending on the requirements of the hydroxyl protecting group. The isocyante forming step is carried out as described above. A second aspect of the invention is a compound of formula (IV) (Figure Removed), wherein R6, R7, R8, R9, R10 and R12 are as defined in the first aspect of the invention. A third aspect of the present invention is a compound of formula (VI) : (Figure Removed), wherein R2, R3, R6, R7, RB, R9, R10, P and Q are as defined in the first aspect of the invention, and the dotted lines indicate the optional presence of a double bond between Cl and C2 or C2 and C3. Definitions Masked aldehyde group (-CPQ): The term masked aldehyde group pertains to a group which can be chemically converted into an aldehyde group, and includes in particular, a protected alcohol group (-CH2OProt), wherein the alcohol protecting group is orthogonal to the nitrogen protecting group, acetal groups (-C(ORac)2) ,thioacetal groups (-C(SRac)2), where each RBC can be independently selected from C1-10 alkyl or together can be a C1-3 alkylene group. Alcohol Protecting Group: This term pertains to a group which is removable leave an alcohol group, without affecting the remainder of the compound to which the alcohol group is attached. Of particular interest are ethers and esters, numerous examples of which are described in Greene and Wuts, "Protective Groups in Organic Synthesis", 3rd edition, John Wiley & Sons (1999), which is incorporated herein by reference. Examples include ethers, in particular silyl ethers (e.g. tert-butyl-dimethyl-silyl ether), esters, for examples acetates and carbonates. Orthogonal: Protecting groups are orthogonal to one another, if one protecting group can be removed from a molecule in which the other protecting group is present, without the other protecting group being removed. For example, the alcohol protecting group acetate is orthogonal to the nitrogen protecting group Teoc, and the alcohol protecting group TBDMS is orthogonal to the nitrogen protecting group benzyloxy carbamate. Nitrogen Protecting Group: This term pertains to a group which is removable from the N10 position of the PBD moiety to leave an N10-C11 imine bond. This application is only concerned with those nitrogen protecting groups which are carbamate, i.e. those which have the structure: The nature of R10 can vary widely, and is chosen dependent on the conditions by which the whole group is eliminated from the molecule. A large number of suitable groups are described in Greene and Wuts, ""Protective Groups in Organic Synthesis", 3rd edition, John Wiley & Sons (1999), which is incorporated herein by reference. These groups include many well know groups such as fmoc (9-fluorenylmethylcarbamate), Troc (2,2,2-trichloroethyl carbamate) and Alloc (allyl carbamate) which can be removed under varying conditions. Also described are carbamate based nitrogen protecting groups which can be cleaved photolytically, such as o-nitrobenzyl carbamate and Nvoc (6-nitroveratryl carbamate).lso of interest are groups which can be cleaved by the action of enzymes. These include the following: roup Removab(Figure Removed) These groups, and others, are described in, for example, Jungheim, L. and Shepherd, T., Chem. Rev., 94, 1553-1566 (1994), WO 00/64864 and Niculescu-Duvaz, D., et al.f J. Med. Chem., 41, 5 5297-5309 (1998). Further enzyme labile groups include the following: (Figure Removed)Prodrugs with such protecting groups are also reviewed Dubowchik, G.M. and Walker, M.A., Pharmacology & Therapeutics, 83, 67-123 (1999). A number of nitrogen protecting groups (including some of the enzyme labile groups discussed above) can be classed as containing a Aself-immolative linker'. Such groups usually has a structure: where Y is NH or 0, n is 0 to 3 and R'10 is such that the whole moiety is a nitrogen protecting group (and may be defined as for R10 below). The self-immolative linker will allow for release of the protected compound when a remote site is activated, proceeding along the lines shown below (for n=0): (Figure Removed) represents a resin bead. The R10 group can be described as an optionally substitued Ca_30 alkyl group, C3-30 heterocylyl group or a C5_30 aryl group or a divalent version of one of these groups linked to another moiety. It is preferred that R10 is not a silane group or is substituted by a silyl group (e.g. -CH2-Si (Me) 3) . The present invention allows the synthesis of N-10 protected PBD compounds where the appropriate chloroformate is not available or is unstable. This for example applies to the Moz (methoxybenzyl carbamate) group which cannot be introduced via the chloroformate as it is too unstable, and other derivatives such as Moz-ON are not sufficiently active to react with a PBD-precursor aniline group. However, the p-methoxybenzyl alcohol is commercially available and reacts smoothly with the isocyanate to give the Moz carbamate (see examples, compound 17). The case of the Teoc carbamate reinforces the point; again the required chloroformate is not very stable and not commercially available. A p-nitrophenyl carbonate derivative of Teoc is available but, again, is not sufficiently active to protect an aromatic amine such as compound 5. Trimethylsilylethanol is commercially available and reacts with isocyanates to afford the Teoc carbamate in good yields (see examples, compound 18). The method of the invention is particularly useful for complex protecting groups, such as those shown in the examples as alcohols 35 and 42. Such protecting groups are required to prepare PBD prodrugs for ADEPT approaches to cancer chemotherapy, and the alcohols are not commercially available and give rise to extremely unstable chloroformates. However, these alcohols react with isocyanates to furnish carbamates in moderate to good yields. Alkyl: The term "alky!" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a hydrocarbon compound having from I to 20 carbon atoms (unless otherwise specified), which may be aliphatic or alicyclic, and which may be saturated or unsaturated (e.g., partially unsaturated, fully unsaturated). Thus, the term "alkyl" includes the sub-classes alkenyl, alkynyl, cycloalkyl, cycloalkyenyl, cylcoalkynyl, etc., discussed below. In the context of alkyl groups, the prefixes (e.g., C1-4, C1-7, C1-20, C2-7, C3-7, etc.) denote the number of carbon atoms, or range of number of carbon atoms. For example, the terra "C1-4 alkyl" as used herein, pertains to an alkyl group having from 1 to 4 carbon atoms. Examples of groups of alkyl groups include C1-4 alkyl ("lower alkyl"), C1-7 alkyl, C1-20 alkyl and C1-30 alkyl. Note that the first prefix may vary according to other limitations; for example, for unsaturated alkyl groups, the first prefix must be at least 2; for cyclic and branched alkyl groups, the first prefix must be at least 3; etc. amples of saturated alkyl groups include, but are not limited to, methyl (C1) , ethyl (C2) , propyl (C3) , butyl (C4), pentyl (C5) , hexyl (C6), heptyl (C7) , octyl (C8) , nonyl (C9) , decyl (C10) , undecyl (C11) , dodecyl (C12) , tridecyl (C13) , tetradecyl (C14) , pentadecyl (C15), and eicodecyl (C20) • Examples of saturated linear alkyl groups include, but are not limited to, methyl (C1), ethyl (C2) , n-propyl (C3) , n-butyl (C4) , n-pentyl (amyl) (C5) , n-hexyl (C6) , and n-heptyl (C7) . Examples of saturated branched alkyl groups include iso-propyl (C3), iso-butyl (C4) , sec-butyl (C4) , tert-butyl (C4) , iso-pentyl (C5) , and neo-pentyl (C5) . Alkenyl: The term "alkenyl" as used herein, pertains to an alkyl group having one or more carbon-carbon double bonds. Examples of groups of alkenyl groups include C2-4 alkenyl, C2-7 alkenyl, C2-20 alkenyl. Examples of unsaturated alkenyl groups include, but are not limited to, ethenyl (vinyl, -CH=CH2), 1-propenyl (-CH=CH-CH3) , 2-propenyl (allyl, -CH-CH=CH2), isopropenyl (1-methylvinyl, -C(CH3)=CH2), butenyl (C4) , pentenyl (C5), and hexenyl (C6) . Alkynyl: The term "alkynyl" as used herein, pertains to an alkyl group having one or more carbon-carbon triple bonds. Examples of groups of alkynyl groups include C2_4 alkynyl, C2.7 alkynyl, C2.20 alkynyl. Examples of unsaturated alkynyl groups include, but are not limited to, ethynyl (ethinyl, -C=CH) and 2-propynyl {propargyl, -CH2-CsCH). Cycloalkyl: The term "cycloalkyl" as used herein, pertains to an alkyl group which is also a cymoiety obtained by removing a hydrogen atom from an alicyclic ring atom of a carbocyclic ring of a carbocyclic compound, which carbocyclic ring may be saturated or unsaturated (e.g., partially unsaturated, fully unsaturated), which moiety has from 3 to 20 carbon atoms (unless otherwise specified), including from 3 to 20 ring atoms. Thus, the term "cycloalkyl" includes the sub-classes cycloalkyenyl and cycloalkynyl. Preferably, each ring has from 3 to 7 ring atoms. Examples of groups of cycloalkyl groups include Ca-ao cycloalkyl, €3-20 cycloalkyl, C3_10 cycloalkyl, C3.7 cycloalkyl. Examples of cycloalkyl groups include, but are not limited to, those derived from: saturated monocyclic hydrocarbon compounds: cyclopropane (C3), cyclobutane (C4) , cyclopentane (C5) , cyclohexane (Ce), cycloheptane (C7) , methylcyclopropane (C^) , dimethylcyclopropane (C5) , methylcyclobutane (C5) , dimethylcyclobutane (Ce) , methylcyclopentane (C6) , dimethylcyclopentane (C7) , methylcyclohexane (C7) , dimethylcyclohexane (Ce) , menthane (C10) ; unsaturated monocyclic hydrocarbon compounds: cyclopropene (C3) , cyclobutene (C saturated polycyclic hydrocarbon compounds: thujane (C10) , carane (Cio) , pinane (Cjo) / bornane (Cio) , norcarane (C?) , norpinane (C7) , norbornane (C7) , adamantane (C10) , decalin (decahydronaphthalene) (CJO); unsaturated polycyclic hydrocarbon compounds: camphene (Ci0) / limonene (Cio) , pinene (Cio); polycyclic hydrocarbon compounds having an aromatic ring: indene (C9) , indane (e.g., 2,3-dihydro-lH-indene) (Cg) , tetraline (1,2,3,4-tetrahydronaphthalene) (C10), acenaphthene (Ci2), fluorene (Ci3), phenalene (C13), acephenanthrene (Ci5), aceanthrene cholanthrene (C2o) . Heterocyclyl: The term "heterocyclyl" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a ring atom of a heterocyclic compound, which moiety has from 3 to 20 ring atoms (unless otherwise specified), of which from 1 to 10 are ring heteroatoms. Preferably, each ring has from 3 to 7 ring atoms, of which from 1 to 4 are ring heteroatoms. In this context, the prefixes (e.g., C3_2o, C3-7, C5-6, etc.) denote the number of ring atoms, or range of number of ring atoms, whether carbon atoms or heteroatoms. For example, the term "Cs-e heterocyclyl" as used herein, pertains to a heterocyclyl group having 5 or 6 ring atoms. Examples of groups of heterocyclyl groups include C3-30 heterocyclyl, C3-20 heterocyclyl, 05-20 heterocyclyl, C3_iS heterocyclyl, C5-15 heterocyclyl, C3_12 heterocyclyl, C5_i2 heterocyclyl, C3.10 heterocyclyl, C5_i0 heterocyclyl, C3-7 heterocyclyl, 05.7 heterocyclyl, and C5.6 heterocyclyl. Examples of monocyclic heterocyclyl groups include, but are not limited to, those derived from: NI: aziridine (C3), azetidine (C4) , pyrrolidine (tetrahydropyrrole) (C5) , pyrroline (e.g., 3-pyrroline, 2,5-dihydropyrrole) (C5) , 2H-pyrrole or 3H-pyrrole (isopyrrole, isoazole) (C5) , piperidine (C6), dihydropyridine (C6) , tetrahydropyridine (C6), azepine (C7) ; Oj.: oxirane (Ca) , oxetane (C4) , oxolane (tetrahydrofuran) (C5) , oxole (dihydrofuran) (C5), oxane (tetrahydropyran) (C6), dihydropyran (C6) , pyran (Ce) , oxepin (C7) ; Si: thiirane (C3) , thietane (C4), thiolane (tetrahydrothlophene) (C5) , thiane (tetrahydrothiopyran) (C6), thiepane (C7) ; Oz' dioxolane (C5) , dioxane (C6) , and dioxepane (C7) ; 03: trioxane (C6); N2: imidazolidine (C5) , pyrazolidine (diazolidine) (C5), imidazoline (C5) , pyrazoline (dihydropyrazole) (C5) , piperazine (C6) ; NjOi: tetrahydrooxazole (C5) , dihydrooxazole (C5) , tetrahydroisoxazole (Cs) , dihydroisoxazole (C5) , morpholine (Ce) tetrahydrooxazine (C6) , dihydrooxazine (Ce) , oxazine (C6) ; NiSi: thiazoline (C5) , thiazolidine (C5) , thiomorpholine (C6) ; N20i: oxadiazine (0$) ; oxathiole (C5) and oxathiane (thioxane) (Ce) / and, j: oxathiazine (C6) . Examples of substituted raonocyclic heterocyclyl groups include those derived from saccharides, in cyclic form, for example, furanoses (Cb) , such as arabinofuranose, lyxofuranose, ribofuranose, and xylofuranse, and pyranoses (C6) , such as allopyranose, altropyranose, glucopyranose, mannopyranose, gulopyranose, idopyranose, galactopyranose, and talopyranose. Aryl: The term "aryl" as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from an aromatic ring atom of an aromatic compound, which moiety has from 3 to 20 ring atoms (unless otherwise specified) . Preferably, each ring has from 5 to 7 ring atoms . In this context, the prefixes (e.g. C3-20, Cb_7, C5-6, etc.) denote the number of ring atoms, or range of number of ring atoms, whether carbon atoms or heteroatoms . For example, the term "C5.6aryl" as used herein, pertains to an aryl group having 5 or 6 ring atoms. Examples of groups of aryl groups include 03-30 aryl, C3-20 aryl, C5-20 aryl, C5-15 aryl, C5.a2 aryl, C5-io aryl, C5.7 aryl, C5.6 aryl, C5 aryl, and C6aryl. The ring atoms may be all carbon atoms, as in "carboaryl groups". Examples of carboaryl groups include C3_2o carboaryl, C5_20 carboaryl, C5_15 carboaryl, C5_12 carboaryl, C5.i0 carboaryl, C5_7 carboaryl, C5_6 carboaryl and Ce carboaryl. Examples of carboaryl groups include, but are not limited to, those derived from benzene (i.e., phenyl) (C6) , naphthalene (Ci0) , azulene (C10) , anthracene (Ci«), phenanthrene (Ci Examples of aryl groups which comprise fused rings, at least one of which is an aromatic ring, include, but are not limited to, groups derived from indane (e.g., 2, 3-dihydro-lH-indene) (C9) , indene (C9) , isoindene (C9) , tetraline (1,2,3,4-tetrahydronaphthalene (Ci0) , acenaphthene (C12) , fluorene (Ci3) , phenalene (Ci3) , acephenanthrene (C15) , and aceanthrene (C16) . Alternatively, the ring atoms may include one or more heteroatoms, as in "heteroaryl groups". Examples of heteroaryl groups include C3.2o heteroaryl, C5-20 heteroaryl, C5-15 heteroaryl, C5-12 heteroaryl, C5.10 heteroaryl, C5-7 heteroaryl, C5_6 heteroaryl, C5 heteroaryl, and C6 heteroaryl . Examples of monocyclic heteroaryl groups include, but are not limited to, those derived from: . pyrrole (azole) (C5) , pyridine (azine) (C6) ; Oa- furan (oxole) (C5) ; j.: thiophene (thiole) (C5) ; oxazole (C5) , isoxazole (Cs) , isoxazine (C6) ; oxadiazole (furazan) (C5) ; N301: oxatriazole (C5) ; N3S1: thiazole (C5) , isothiazole (C5) ; N2: imidazole (1, 3-diazole) (C5) , pyrazole (1, 2-diazole) (Cs) , pyridazine (1,2-diazine) (C6) / pyrimidine (1, 3-diazine) (C6) (e.g., cytosine, thyraine, uracil), pyrazine (1, 4-diazine) (C6) ; N3: triazole (C5) , triazine (C6) ; and, N4: tetrazole (C5) . Examples of heteroaryl groups which comprise fused rings, include, but are not limited to: Cg heteroaryl groups (with 2 fused rings) derived from benzofuran (O1) , isobenzofuran (O1) , indole (N1) , isoindole indolizine (Ni) , indoline (Na), isoindoline (Nj) , purine (N4) (e.g., adenine, guanine), benzimidazole (N2) , indazole (N2) , benzoxazole (Nidi) , benzisoxazole (NjOj) , benzodioxole (02), benzofurazan (N^) , benzotriazole (N3) , benzothiofuran (Si), benzothiazole (NiSi) , benzothiadiazole (N2S); Cio heteroaryl groups (with 2 fused rings) derived from chromene (O^) , isochromene (Oi) , chroman (Oi) , isochroman (Oi) / benzodioxan (02) , guinoline (Na), isoquinoline (Nj) , quinolizine (Ni) , benzoxazine (NiOa) , benzodiazine (N2) , pyridopyridine (N2) , quinoxaline (N2) , quinazoline (N2) , cinnoline (N2), phthalazine (N2) , naphthyridine (N2), pteridine (N GXJ. heteroaryl groups (with 2 fused rings) derived from benzodiazepine (N2) ; Cis heteroaryl groups (with 3 fused rings) derived from carbazole (Nj) , dibenzofuran (Oj), dibenzothiophene (Sa), carboline (N2) , perimidine (N2) , pyridoindole (N2) ; and, Cn heteroaryl groups (with 3 fused rings) derived from acridine (Nx) , xanthene (Oi) , thioxanthene (Si), oxanthrene (02) , phenoxathiin (OaSi) , phenazine (N2) , phenoxazine (NiOa) , phenothiazine (NjSj), thianthrene (S2) , phenanthridine (Nj), phenanthroline (N2) , phenazine (N2) . Heteroaryl groups which have a nitrogen ring atom in the form of an -NH- group may be N-substituted, that is, as -NR-. For example, pyrrole may be N-methyl substituted, to give N-methylpyrrole. Examples of N-substitutents include, but are not limited to C1-7 alkyl, C3-20 heterocyclyl, C5-20 aryl, and acyl groups. Heterocyclic groups (including heteroaryl groups) which have a nitrogen ring atom in the form of an -N= group may be substituted in the form of an N-oxide, that is, as -N(→0)= (also denoted -N+{→0-)=). For example, quinoline may be substituted to give quinoline N-oxide; pyridine to give pyridine N-oxide; benzofurazan to give benzofurazan N-oxide (also known as benzofuroxan). Cyclic groups may additionally bear one or more oxo (=0) groups on ring carbon atoms. The above groups, whether alone or part of another substituent, may themselves optionally be substituted with one or more groups selected from themselves and the additional substituents listed below. Halo: -F, -Cl, -Br, and -I. Hydroxy: -OH. Ether: -OR, wherein R is an ether substituent, for example, a Ci_7 alkyl group (also referred to as a Ca_7 alkoxy group, discussed below), a C3_20 heterocyclyl group (also referred to as a C3_2o heterocyclyloxy group) , or a C5_20 aryl group (also referred to as a C5-20 aryloxy group), preferably a Chalky! group. Alkoxy: -OR, wherein R is an alkyl group, for example, a C1-7 alkyl group. Examples of Ca-7 alkoxy groups include, but are not limited to, -OMe (methoxy), -OEt (ethoxy), -O(nPr) (n-propoxy), -O(iPr) (isopropoxy), -O(nBu) (n-butoxy), -O(sBu) (sec-butoxy), -O(iBu) (isobutoxy), and -0{tBu) (tert-butoxy). Acetal: -CHfOR1) (OR2), wherein R1 and R2 are independently acetal substituents, for example, a C1-7 alkyl group, a C3-20 heterocyclyl group, or a Cs-2o aryl group, preferably a C1-7 alkyl group, or, in the case of a "cyclic" acetal group, R1 and R2, taken together with the two oxygen atoms to which they are attached, and the carbon atoms to which they are attached, form a heterocyclic ring having from 4 to 8 ring atoms. Examples of acetal groups include, but are not limited to, -CH(OMe)2, -CH(OEt)2, and -CH(OMe)(OEt). Hemiacetal: -CH(OH)(OR1), wherein R1 is a hemiacetal substituent, for example, a C1-7 alkyl group, a C3-20 heterocyclyl group, or a C5-20 aryl group, preferably a C1-7 alkyl group. Examples of hemiacetal groups include, but are not limited to, -CH(OH)(OMe) and -CH(OH)(OEt). Ketal: -CR(OR1) (OR2), where Ra and R2 are as defined for acetals, and R is a ketal substituent other than hydrogen, for example, a C1-7 alkyl group, a C3-20 heterocyclyl group, or a C5_20 aryl group, preferably a C1-7 alkyl group. Examples ketal groups include, but are not limited to, -C(Me) (OMe)2, -C(Me) (OEt)2, -C(Me) (OMe) (OEt), -C(Et) (OMe)2, -C(Et) (OEt)2, and -C(Et) (OMe) (OEt) . Hemiketal: -CR(OH) (OR1) , where R1 is as defined for hemiacetals, and R is a hemiketal substituent other than hydrogen, for example, a C1-7 alkyl group, a C3-2 heterocyclyl group, or a C5-200 aryl group, preferably a alkyl group. Examples of hemiacetal groups include, but are not limited to, -C(Me)(OH)(OMe), -C(Et)(OH)(OMe), -C(Me)(OH)(OEt), and -C(Et)(OH)(OEt). Oxo (keto, -one): =0. Thione (thioketone): =S. Imino (imine): =NR, wherein R is an imino substituent, for example, hydrogen, alkyl group, a C3_;0 heterocyclyl group, or a Cs-2o aryl group, preferably hydrogen or a Ci_7 alkyl group. Examples of ester groups include, but are not limited to, =NH, =NMe, =NEt, and =NPh. Formyl (carbaldehyde, carboxaldehyde): -C(=0)H. •20 20 Acyl (keto): -C(=O)R, wherein R is an acyl substituent, for example, a Ci-7 alkyl group (also referred to as Ci_7 alkylacyl or Ci-7 alkanoyl) , a C3-2o heterocyclyl group (also referred to as C3-heterocyclylacyl) , or a C5.20 aryl group (also referred to as C5. arylacyl), preferably a CI-T alkyl group. Examples of acyl groups include, but are not limited to, -C( Carboxy (carboxylic acid): -C(=0)OH. Thiocarboxy (thiocarboxylic acid): -C(=S)SH. Thiolocarboxy (thiolocarboxylic acid): -C(=0)SH. Thionocarboxy (thionocarboxylic acid): -C(=S)OH. Imidic acid: -C(=NH)OH. Hydroxamic acid: -C(=NOH)OH. Ester (caxboxylate, carboxylic acid ester, oxycarbonyl): -C(=O)OR, wherein R is an ester substituent, for example, a Ci--j alkyl group, a C3-2o heterocyclyl group, or a C5.2o aryl group, preferably a Ca_7 alkyl group. Examples of ester groups include, but are not limited to, -C(=O)OCH3, -C(=0)OCH2CH3, -C (=0) OC (CH3)3, and -C(=0)OPh. Acyloxy (reverse ester): -OC(=O)R, wherein R is an acyloxy substituent, for example, a Cj-7 alkyl group, a C3-2o heterocyclyl group, or a C5.2o aryl group, preferably a Ci-7 alkyl group. Examples of acyloxy groups include, but are not limited to, -OC(=0)CH3 (acetoxy) , -OC (=0) CH2CH3, -OC (=0) C (CH3) 3, -OC(=0)Ph, and -OC(=O)CH2Ph. Oxycarboyloxy: -OC(=0)OR, wherein R is an ester substituent, for example, a Ci-7 alkyl group, a C3_20 heterocyclyl group, or a C5-20 aryl group, preferably a Ci_7 alkyl group. Examples of ester groups include, but are not limited to, -OC(=0)OCH3, -OC(=0)OCH2CH3, -OC(=0)OC(CH3)3, and -OC(=0)OPh. Amino: -NRJR2, wherein R3 and Rz are independently amino substituents, for example, hydrogen, a CI-T alkyl group (also referred to as Ci_7 alkylamino or di-Ci-v alkylamino') , a C3-20 heterocyclyl group, or a C5_20 aryl group, preferably H or a CI_T alkyl group, or, in the case of a "cyclic" amino group, R1 and R2, taken together with the nitrogen atom to which they are attached, form a heterocyclic ring having from 4 to 8 ring atoms. Amino groups may be primary (-NH2) , secondary (-NHR1) , or tertiary (-NHR1RZ) , and in cationic form, may be quaternary (-+NR1R2R3) . Examples of araino groups include, but are not limited to, -NH2, -NHCH3, -NHC(CH3)2/ -N(CH3)2, -N(CH2CH3)2, and -NHPh. Examples of cyclic amino groups include, but are not limited to, aziridino, azetidino, pyrrolidino, piperidino, piperazino, morpholino, and thiomorpholino. Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide): -C(=0)NR1R2, wherein R1 and R2 are independently amino substituents, as defined for amino groups. Examples of amido groups include, but are not limited to, -C(=0)NH2, -C(=0)NHCH3, -C(=0)N(CH3)2, -C(=0)NHCH2CH3, and -C (=0) N (CH2CH3) 2, as well as amido groups in which R1 and R2, together with the nitrogen atom to which they are attached, form a heterocyclic structure as in, for example, piperidinocarbonyl, morpholinocarbonyl, thiomorpholinocarbonyl, and piperazinocarbonyl. Thioamido (thiocarbamyl) : -C(=S)NR1R2, wherein R1 and R2 are independently amino substituents, as defined for amino groups. Examples of amido groups include, but are not limited to, -C(=S)NH2, -C(=S)NHCH3, -C (=S) N (CH3) 2, and -C (=S) NHCH2CH3. Acylamido (acylamino) : -NR1C(=0)R2, wherein R1 is an amide substituent, for example, hydrogen, a Ci--? alkyl group, a C3.20 heterocyclyl group, or a Cs-2o aryl group, preferably hydrogen or a Ci-7 alkyl group, and R2 is an acyl substituent, for example, a Ci-v alkyl group, a C3-2o heterocyclyl group, or a C6.20aryl group, preferably hydrogen or a Cj-7 alkyl group. Examples of acylamide groups include, but are not limited to, -NHC(=0)CH3 , -NHC(=0)CH2CH3, and -NHC(=0)Ph. R1 and R2 may together form a cyclic structure, as in, for example, succinimidyl, maleimidyl, and (Figure Removed) Aminocarbonyloxy: -OC(=0)NR1R2, wherein R1 and R2 are independently amino substituents, as defined for amino groups. Examples of aminocarbonyloxy groups include, but are not limited to, -OC(=0)NH2/ -OC(=0)NHMe, -OC(=0)NMe2, and -OC(=0)NEt2. Ureido: -N (R1) CONR2R3 wherein Rz and R3 are independently amino substituents, as defined for amino groups, and R1 is a ureido substituent, for example, hydrogen, a Ci_7 alkyl group, a C3_2o heterocyclyl group, or a C5-20 aryl group, preferably hydrogen or a Ci--? alkyl group. Examples of ureido groups include, but are not limited to, -NHCONH2, -NHCONHMe, -NHCONHEt, -NHCONMe2, -NHCONEt2, -NMeCONH2, -KMeCONHMe, -NMeCONHEt, -NMeCONMe2/ and -NMeCONEt2. Guanidino: -NH-C(=NH)NH2. Imino: =NR, wherein R is an imino substituent, for example, for example, hydrogen, a CI_T alkyl group, a C3_20 heterocyclyl group, or a C5-20 aryl group, preferably H or a Ci_7alkyl group. Examples of imino groups include, but are not limited to, =NH, =NMe, and =NEt. Amidine (amidino): -C(=NR)NR2, wherein each R is an amidine substituent, for example, hydrogen, a Cj-7 alkyl group, a C3_20 heterocyclyl group, or a C5-20 aryl group, preferably H or a Ci-7 alkyl group. Examples of amidine groups include, but are not limited to, -C(=NH)NH2, -C(=NH)NMe2, and -C (=NMe) NMe2. Nitro: -N02. Nitroso: -NO. Azido: -N3. Cyano (nitrile, carbonitrile): -CN. Isocyano: -NC. Cyanato: -OCN. Isocyanato: -NCO. Thiocyano (thiocyanato): -SCN. Isothiocyano (isothiocyanato): -NCS. Sulfhydryl (thiol, mercapto): -SH. Thioether (sulfide): -SR, wherein R is a thioether substituent, for example, a CI_T alkyl group (also referred to as a Ca_7alkylthio group), a C3_20 heterocyclyl group, or a C5-20 aryl group, preferably a d_7 alkyl group. Examples of Ci_7 alkylthio groups include, but are not limited to, -SCH3 and -SCH2CH3. Disulfide: -SS-R, wherein R is a disulfide substituent, for example, a Cj.v alkyl group, a C3_20 heterocyclyl group, or a €5-20 aryl group, preferably a Ci_7 alkyl group (also referred to herein as Ca_7 alkyl disulfide) . Examples of Ca_7 alkyl disulfide groups include, but are not limited to, -SSCH3 and -SSCH2CH3. Sulfine (sulfinyl, sulfoxide): -S(=0)R, wherein R is a sulfine substituent, for example, a Ci-7 alkyl group, a C3.20 heterocyclyl group, or a Cs-zo aryl group, preferably a Ci_7 alkyl group. Examples of sulfine groups include, but are not limited to, -S(=0)CH3 and -S(=0)CH2CH3. Sulfone (sulfonyl): -S(=0);R, wherein R is a sulfone substituent, for example, a Ci-? alkyl group, a C3.2o heterocyclyl group, or a C5-2o aryl group, preferably a Ca.7 alkyl group, including, for example, a fluorinated or perfluorinated Ca~7 alkyl group. Examples of sulfone groups include, but are not limited to, -S(=0)2CH3 (methanesulfonyl, mesyl), -S(=0)ZCF3 (triflyl), -S(=0)2CH2CH3 (esyl), -S(=O)2C4F9 (nonaflyl) , -S (=0)2CH2CF3 (tresyl) , -S(=0)2CH2CH2NH2 (tauryl), -S(=0)2Ph (phenylsulfonyl, besyl) , 4-methylphenylsulfonyl (tosyl), 4-chlorophenylsulfonyl (closyl), 4-bromophenylsulfonyl (brosyl), 4-nitrophenyl (nosyl), 2-naphthalenesulfonate (napsyl), and 5-dimethylamino-naphthalen-1-ylsulfonate (dansyl). Sulfinic acid (sulfino): -S(=0)OH, -S02H. Sulfonic acid (sulfo): -S(=0)2OH, -S03H. Sulfinate (sulfinic acid ester): -S(=0)OR; wherein R is a sulfinate substituent, for example, a Ci._7 alkyl group, a C3-20 heterocyclyl group, or a Cs_20 aryl group, preferably a Ci_7 alkyl group. Examples of sulfinate groups include, but are not limited to, -S(=0)OCH3 (methoxysulfinyl; methyl sulfinate) and -S(=0)OCH2CH3 (ethoxysulfinyl; ethyl sulfinate). Sulfonate (sulfonic acid ester): -S(=0)2OR, wherein R is a sulfonate substituent, for example, a Cj-7 alkyl group, a C3-20 heterocyclyl group, or a C5-20 aryl group, preferably a Ca-7 alkyl group. Examples of sulfonate groups include, but are not limited to, -S(=0)2OCH3 (methoxysulfonyl; methyl sulfonate) and -S(=0)2OCH2CH3 (ethoxysulfonyl; ethyl sulfonate). Sulfinyloxy: -OS(=0)R, wherein R is a sulfinyloxy substituent, for example, a Ca_7 alkyl group, a C3-20 heterocyclyl group, or a C5-2o aryl group, preferably a Ca-7 alkyl group. Examples of sulfinyloxy groups include, but are not limited to, -OS(=0)CH3 and -OS(=0)CH2CH3. Sulfonyloxy: -OS(=0)2R, wherein R is a sulfonyloxy substituent, for example, a Ci_7 alkyl group, a C3_20 heterocyclyl group, or a Cs-so aryl group, preferably a Ci_7 alkyl group. Examples of sulfonyloxy groups include, but are not limited to, -OS(=0)2CH3 (mesylate) and -OS (=0)2CH2CH3 (esylate) . Sulfate: -OS(=0)2OR; wherein R is a sulfate substituent, for example, a Ci_7 alkyl group, a C3-20 heterocyclyl group, or a C5-2o aryl group, preferably a Ca-7 alkyl group. Examples of sulfate groups include, but are not limited to, -OS(=0)20CH3 and -SO(=0)2OCH2CH3. Sulfamyl (sulfamoyl; sulfinic acid amide; sulfinamide): -S (=0)NR1R2, wherein R1 and R2 are independently amino substituents, as defined for amino groups. Examples of sulfamyl groups include, but are not limited to, -S(=0)NH2, -S(=0)NH(CH3) , -S(=0)N(CH3)2, -S(=0)NH(CH2CH3), -S (=0) N (CH2CH3) 2, and -S (=0) NHPh. Sulfonamido (sulfinamoyl; sulfonic acid amide; sulfonamide): -S (=0)2NR1R2, wherein R1 and R2 are independently amino substituents, as defined for amino groups. Examples of sulfonamido groups include, but are not limited to, -S(=0)2NH2, -S(=0)2NH(CH3) , -S(0)2N(CH3}2, -S (=O) 2NH (CH2CH3) , -S (=0) 2N (CH2CH3) 2, and -S(=0)2NHPh. Sulfaraino: -NR^ (=0) 2OH, wherein R1 is an amino substituent, as defined for amino groups. Examples of sulfamino groups include, but are not limited to, -NHS(=0)2OH and -N (CH3) S (=0) 2OH. Sulfonamino: -NR1S(=0)2R, wherein R1 is an amino substituent, as defined for amino groups, and R is a sulfonamino substituent, for example, a Ci_7 alkyl group, a C3-20 heterocyclyl group, or a C5-20 aryl group, preferably a Ci_7 alkyl group. Examples of sulfonamino groups include, but are not limited to, -NHS(=0)2CH3 and -N(CH3)S(=0)2C6HS. Sulfinamino: -NR1S(=0)R, wherein R1 is an amino substituent, as defined for amino groups, and R is a sulfinamino substituent, for example, a Ci_7 alkyl group, a C3-2o heterocyclyl group, or a C5.20 aryl group, preferably a Ci-7 alkyl group. Examples of sulfinamino groups include, but are not limited to, -NHS(=0)CH3 and -N(CH3)S(=0)C6H5. Phosphino (phosphine): -PR2, wherein R is a phosphino substituent, for example, -H, a d_- alkyl group, a C3-zo heterocyclyl group, or a C5-20 aryl group, preferably -H, a Ci-7 alkyl group, or a C5_20 aryl group. Examples of phosphino groups include, but are not limited to, -PH2, -P(CH3)2, -P(CH2CH3)2, -P(t-Bu)2, and -P(Ph)2. Phospho: -P(=0)2. Phosphinyl (phosphine oxide): -P(=O)R2, wherein R is a phosphinyl substituent, for example, a Ci-7 alkyl group, a C3.20 heterocyclyl group, or a C5.20 aryl group, preferably a Ca_7 alkyl group or a C5_2o aryl group. Examples of phosphinyl groups include, but are not limited to, -P(-O) (CH3)8, -P (=0) (CH2CH3) 2, -P(=0) (t-Bu)2, and -P(=0)(Ph)2. Phosphonic acid (phosphono): -P(=0)(OH)2. Phosphonate (phosphono ester): -P(=0)(OR)2, where R is a phosphonate substituent, for example, -H, a Cj_7 alkyl group, a C3_2o heterocyclyl group, or a C5-20 aryl group, preferably -H, a Ci_7 alkyl group, or a C5-20 aryl group. Examples of phosphonate groups include, but are not limited to, -P(=0) (OCH3)2, -P(=0) (OCH2CH3)2, -P(=0)(0-t-Bu)2, and -P(=O)(OPh)2. Phosphoric acid (phosphonooxy): -OP(=0)(OH)2. Phosphate (phosphonooxy ester): -OP(=0)(OR)2, where R is a phosphate substituent, for example, -H, a Ci-7 alkyl group, a C3-20 heterocyclyl group, or a C5_2o aryl group, preferably -H, a Ca-7 alkyl group, or a C6_20 aryl group. Examples of phosphate groups include, but are not limited to, -OP(=0) (OCH3)2, -OP(=0) (OCH2CH3) 2, -OP(=0)(0-t-Bu)2, and ~OP(=0)(OPh)2. Phosphorous acid: -OP(OH)2. Phosphite: -OP(OR)2, where R is a phosphite substituent, for example, -H, a Ci-7 alkyl group, a C3.2o heterocyclyl group, or a C5-2o aryl group, preferably -H, a Ci-7 alkyl group, or a C5_20 aryl group. Examples of phosphite groups include, but are not limited to, -OP(OCH3)2, -OP(OCH2CH3)2, -OP(0-t-Bu)2, and -OP (OPh) 2. Phosphoramidite: -OP (OR1) -NR22, where R1 and R2 are phosphoramidite substituents, for example, -H, a (optionally substituted) Ci_7 alkyl group, a C3_20 heterocyclyl group, or a C5.20 aryl group, preferably -H, a Cj_7 alkyl group, or a C5_2o aryl group. Examples of phosphoramidite groups include, but are not limited to, -OP(OCH2CH3)-N(CH3)2, -OP(OCH2CH3)-N{i-Pr)2, and -OP (OCH2CH2CN)-N (i-Pr)2. Phosphoramidate: -OP(=0) (OR1)-NR22, where R1 and R2 are phosphoramidate substituents, for example, -H, a (optionally substituted) Cj_7 alkyl group, a C3_20 heterocyclyl group, or a C5-20 aryl group, preferably -H, a Ci_v alkyl group, or a C6-20 aryl group. Examples of phosphoramidate groups include, but are not limited to, -OP(=0) (OCH2CH3)-N(CH3)2, -OP (=0) (OCH2CH3) -N (i-Pr) 2, and -OP(=0) (OCH2CH2CN)-N(i-Pr)2. Alkylene C3_i2 alkylene: The term "C3-i2 alkylene", as used herein, pertains to a bidentate moiety obtained by removing two hydrogen atoms, either both from the same carbon atom, or one from each of two different carbon atoms, of a hydrocarbon compound having from 3 to 12 carbon atoms (unless otherwise specified), which may be aliphatic or alicyclic, and which may be saturated, partially unsaturated, or fully unsaturated. Thus, the term "alkylene" includes the sub-classes alkenylene, alkynylene, cycloalkylene, etc., discussed below. Examples of linear saturated C3_i2 alkylene groups include, but are not limited to, -(CH2)n- where n is an integer from 3 to 12, for example, -CH2CH2CH2- (propylene) , -CH2CH2CH2CH2- (butylene), -CH2CH2CH2CH2CH2- (pentylene) and -CH2CH2CH2CH2CH2CH2CH;- (heptylene) . Examples of branched saturated C3-i2 alkylene groups include, but are not limited to, -CH(CH3) CH2-, -CH(CH3) CH2CH2-, -CH(CH3)CH2CH2CH2-, -CH2CH (CH3) CH2-, -CH2CH (CH3) CH2CH2-, -CH (CH2CH3)-, -CH(CH2CH3)CH2-, and -CH2CH (CH2CH3) CH2-. Examples of linear partially unsaturated C3-i2 alkylene groups (C3_i2 alkenylene, and alkynylene groups) include, but are not limited to, -CH=CH-CH2-, -CH2-CH=CH2-, -CH=CH-CH2-CH:-, -CH=CH-CH2-CH2-CH2-, -CH=CH-CH=CH-, -CH=CH-CH=CH-CH2-, -CH=CH-CH=CH-CH2-CH2-, -CH=CH-CH2-CH=CH-, -CH=CH-CH2-CH2-CH=CH-, and -CH2-CsC-CH2-. Examples of branched partially unsaturated C3_i2 alkylene groups (C3_i2 alkenylene and alkynylene groups) include, but are not limited to, -C(CH3)=CH-, -C (CH3) =CH-CH2-, ~CH=CH-CH (CH3) - and -CsC-CH(CH3)-. Examples of alicyclic saturated C3-i2 alkylene groups (C3.12 cycloalkylenes) include, but are not limited to, cyclopentylene (e.g. cyclopent-1,3-ylene), and cyclohexylene (e.g. cyclohex-1,4-ylene). Examples of alicyclic partially unsaturated C3_i2 alkylene groups (C3-i2 cycloalkylenes) include, but are not limited to, cyclopentenylene (e.g. 4-cyclopenten-l,3-ylene), cyclohexenylene (e.g. 2-cyclohexen-l,4-ylene; 3-cyclohexen-l,2-ylene; 2,5-cyclohexadien-l,4-ylene). The term "Ca-3 alkylene" as used herein, is defined in a similar manner to the above group, but with from 1 to 3 carbon atoms. Solvents Solvents may conveniently be classified according to one or more of their physical or chemical properties. For example, solvents may be classified according to their polarity, that is, their permanent dipole moment. Examples of highly polar solvents include dimethylformamide (DMF), dimethylacetamide, and acetonitrile (ACN). Examples of moderately polar solvents include acetone, methanol, tetrahydrofuran (THF), ethyl acetate (AcOEt), and water. Examples of relatively non-polar solvents include diethyl ether, chloroform, and dichloromethane (DCM). Examples of non-polar and virtually non-polar solvents include alkanes, benzene, toluene, and carbon tetrachloride. Solvents may also be classified as "protic" or "aprotic" according to their proton-exchange properties. Protic solvents accept and/or donate protons. Examples of protic solvents include water, alcohols, carboxylic acids (e.g., acetic acid), and amines (e.g., ammonia, pyridine). Aprotic solvents neither accept nor donate protons. Examples of aprotic solvents include carbon tetrachloride, chloroform, dichloromethane (DCM), acetonitrile (ACN), ethyl acetate (AcOEt), dimethylacetamide, tetrahydrofuran (THF), dimethylformamide (DMF), toluene, benzene, acetone, ethers (e.g., diethyl ether), alkanes (e.g., hexane), dimethylsulfoxide (DMSO), sulfur dioxide, hexamethylphosphoramide (HMPA), and, .tetramethylurea. Amphoteric solvents, such as water, are capable of both accepting and donating protons. Solvents may be classifed as "hydroxylic" or "non-hydroxylic" according to whether they contain one or more hydorxoyl, i.e. -OH, groups. Hydroxylic solvents include water and alcohols (e.g. methanol, ethanol), and non-hydroxylic solvents include dichoromehtane and toluene. Solvents may also be classified as "organic" or "inorganic" according to their chemical composition. Conventionally, organic solvents comprise, at least, carbon atoms, while inorganic solvents do not. Examples of inorganic solvents include water, ammonia, and sulfur dioxide. Anhydrous solvents are solvents which contain less than 0.1% by weight of water, and preferably less than 0.01% or 0.001% by weight of water. Further Preferences Preferred methods of the invention may include any appropriate combination of method steps as described above. Preferences for the reaction conditions of the method steps of the invention have been described above. Preferences for the various groups defined above are expressed below, and may be combined with each other as appropriate. R2 is preferably selected from R and =CH-R (wherein, in one aspect, R is preferably optionally substituted phenyl). If there is a double bond between Cl and C2 or C2 and C3, R2 is preferably a group of formula X: wherein z is selected from cyano, R, carboxy, ester and amido. R3 R3 is preferably H. Rf R6 is preferably selected from H and Ca_7 alkoxy, and more preferably H and methoxy, with H being the most preferred. R7 R7 is preferably selected from H and OR, where OR is preferably optionally substituted Ca-7 alkoxy (e.g. methoxy, benzyloxy). R8 R8 is preferably selected from either: H and OR, where OR is preferably optionally substituted (^-7 alkoxy (e.g. methoxy, benzyloxy); or is a dimer link. R9 R9 is preferably H. R11 R11 is preferably H, but may be Ci--? alkyl, R12 R12 is preferably methyl. Examples General Experimental Methods The progress of reactions was monitored by thin-layer chromatography (TLC) using Merck grade 7749 silica gel containing binding and fluorescence indicators, on glass plates. Visualization of TLC plate was achieved with UV light, unless otherwise stated. Flash column chromatography was performed using Merck silica gel 60 (0.040-0.063). The majority of organic solvents and reagents used were bought from Fischer, Lancaster and Aldrich Chemical Co. and inorganic drying agents from BDH. 1H and 13C NMR spectra were obtained on a Bruker 250 MHz/52mm; IR spectra were recorded with a FT-IR Spectrometer Spectrum 1000. The optical rotation of compounds was determined at ambient temperature with an ADP 220 polarimeter. Mass Spectrometry was carries out on a Micromass platform using either electrospray or Atmospheric Pressure chemical ionisation. Microanalysis was performed using a Carlo Erba 1108 elemental analyser. Synthesis of monomer isocyanates (i) 12-(tert-Butyl-dimethyl-silanyloxymethyl)-pyrrolidin-1-yl]- (2-isocyanato-4,5-dimethoxy-phenyl)-methanone; and (ii) (2-Hydroxymethyl-pyrrolidin-1-yl)-(2-isocyanato-4,5- dimethoxy-phenyl)-methanone (8) (Figure Removed) (4,5-Dimethoxy-2-nitro-phenyl) - (2-hydroxymethyl-pyrrolidin-l-yl) -methanone (2) (Figure Removed)Oxalyl chloride (9.22 g, 6.4 mL, 72.6 mmol, 1.1 eq) and DMF (cat) were added to a suspension of 4,5-dimethoxy-2-nitrobenzoic acid (1, 15.0 g, 66 rnmol) in anhydrous CH2C12 (150 mL) under a N2 atmosphere. The suspension was stirred at room temperature for 18 hours. The resultant solution was added .dropwise to a solution of (S)-(+)-2-pyrrolidine-methanol (7.33 g, 7.16 mL, 72.6 mmol, 1.1 eq) and triethylamine (14.77 g, 20.21 mL, 145 mmol, 2.2 eq) in anhydrous CH2C12 (100 mL) at -40°C (dry ice/CH3CN) under a N2 atmosphere. The reaction mixture was allowed to come to room temperature and stirred for 18 h. The mixture was washed with 1 M HC1 (3 x 200 mL) , H20 (2 x 200 mL) , satd NaCllaq, (200 mL) , dried (MgS04) and evaporated in vacuo to give a yellow foam. Trituration with 3% MeOH/EtQAc gave a white solid. Filtration and concentration of the filtrate gave a second crop of white solid. Total yield (18.13g, 88.5%). mp 124.4 - 127. 8°C; [c [2- (tert-Butyl-dimethyl-sllanyloxymethyl) -pyrrolidhanone (3)o A solution of t-butyldimethylsichloride (9.04 g, 60 mmol, 1.2 eq), imidazole (8.51 g, 125 mmol, 2.5 eq) and nitro-alcohol 2 (15.55 g, 50 mmol, I eq) in anhydrous DMF (30 mL) was stirred at room temperature under a N2 atmosphere for 16 hours. The reaction mixture was diluted with H20 (500 mL) and extracted with EtOAc (2 x 200 mL). The combined organic extracts were washed with H20 (2 x 200 mL) , satd NaCl(aq) (2 x 200 mL) , dried (MgSO Pyridine (9.03 mL, 111.7 mmol, 1.1 eq) and acetic anhydride (10.5 mL, 111.7 mmol, 1.1 eq) were added to a solution of the nitro-alcohol 2 (31.5 g, 101.5 nvmol, leq) in anhydrous THF (300 mL) under a N2 atmosphere. DMAP (2.48 g, 20.3 mmol, 0.2 eq) was added portion wise and the mixture was stirred at room temperature for 2.5 h. The solvent was removed in vacuo, the residue was treated with 1 M HC1 (300 mL) and extracted with EtOAc (300 mL). The organic extract was washed with 1 M HC1 (2 x 200 mL), H20 (3 x 200 mL), satd NaCl (aq) (2 x 200 mL) , dried (MgS04) and evaporated in vacuo to give the product as a yellow foam (34.5 g, 96%): [a]27D -89.0 (c 1.0, CHC13); XH NMR (CDC13) 6 7.69 (s, 1H) , 6.82 (s, 1H) , 4.05-3.95 (m 9H), 3.24-3.08 (m, 2H), 2.11 (s, 3H) , 2.00-1.41 (m, 4H); 13C NMR (CDC13) 5 170.9, 170.2, 151.7, 141.5, 140.7, 112.5, 111.3, 100.7, 64.6, 56.8, 55.7, 55.5, 49.8, 27.9, 24.7, 20.9; IR (neat) 3629, 3464, 2982, 1751, 1659, 1585; HRMS m/z calcd for C16H2iN207 353.1355 (M + H) , found 353.1349. (2-Amino-4,5-dimethoxy-phenyl)-[2- (tert-butyl-dimethyl-silanyloxymethyl) -pyrrolidin-1-yl]-methanon(Figure Removed)on of the nitro compound 3 (19.8 g, 46.6 mmol) in ethanol (180 mL) was hydrogenated (Parr apparatus) over 10% Palladium on carbon (2 g, 10 wt%), maintaining the H2 pressure at 16 psi. The reaction was complete when no more H2 was consumed. The mixture was filtered through celite and the ethanol evaporated in vacua. Purification by flash column chromatography (2% MeOH/EtOAc) gave the product as a yellow oil (16.8 g, 91%). [a]25D -149.6° (c = 1.0, CHC13); :H NMR (CDC13) 6 6.75 (s, 1H), 6.24 (s, 1H) , 4.45 -4.32 (m, 1H) , 3.84 (s, 3H), 3.78 (s, 3H), 3.64 - 3.50 (m, 4H), 2.08 - 2.00 (m, 3H), 1.93 - 1.74 (m, 1H), 0.89 (s, 9H), 0.09 -0.01 (m, 6H); 13C NMR (CDC13) 6 169.8, 151.8, 141.8, 141.0, 112.5, 100.9, 63.1, 58.7, 56.9, 56.0, 27.5, 26.5, 25.6, 18.5, -5.0; MS (ES+) m/z 395 (M+l); IR (neat) 3465, 3363, 3225, 2973, 2738, 1636, 1523, 1475, 1291 cm'1. (2-Amino-4,5-dimethoxy-phenyl) -[2- (acetoxymethyl) -pyzzolidin-1-yl]-methanone (6) (Figure Removed) solution of the nitro compound 4 (34.0 g, 96.5 iranol) in ethanol (80 mL) was hydrogenated (Parr apparatus) over 10% Palladium on carbon (3.4 g, 10 wt%) at 30 psi for 6 h. The mixture was filtered through celite and the ethanol evaporated in vacuo. Purification by flash column chromatography (2% MeOH/EtOAc) gave the product as a yellow oil (16.2 g, 52%): [a]26D -168.5 (c 1.0, CHC13) ; 1H NMR (CDC13) 6 6.73 (s, IE), 6.26 (s, 1H), 4.66-4.53 (bs, 2H), 4.33-4.16 (bs, 2H), 3.91-3.71 (m, 7H), 3.60-3.44 (m, 2H), 2.22-2.01 (m, 5H), 2.01-1.87 (m, 1H), 1.87-1.70 (m, 3H); 13C NMR 170.9, 167.2, 154.4, 149.2, 137.3, 128.2, 109.2, 107.2, 63.9, 56.7, 56.5, 55.8, 48.4, 27.6, 24.1, 20.6; IR (neat) 3453, 3355, 3240, 2968, 2833, 1731, 1621, 1514; MS (ES+) m/z 323 (M+- + 1). [2-(tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidin-1-yl]- (2-isocyanato-4,5-dimethoxy-phenyl)-methanone (7) solution of triethylamine (1.35 eq.) in anhydrous toluene was added to the amine (5)(1 eq.) and triphosgene (0.36 eq.) in anhydrous toluene under a N2 atmosphere. The reaction was finished after 2 hours, (monitored by IR, vNCo 2265 cm"1) . The product was used without further purification. This was prepared from (6) in the same manner as above for (7) (2-Hydroxymethyl-pyrrolidin-l-yl) - (2-±socyanato-4,5-dimethoxy-phenyl)-methanone (8) Synthesis of protecting group precursors: (i) 2- [3- (4-Hydroxymethyl-phenyl)ureido] -pentanedioic acid diallyl ester (35) (Figure Removed)tert-Butyl-dimethyl- (4-nitro-benzyloxy) -silane (3(Figure Removed) A solution of t-butyldimethylsilyl chloride (8.53 g, 56.3 mmol, 1.3 eq), imidazole (7.38 g, 108.3 mmol, 2.5 eq) and 4-nitro-benzyl alcohol (6.64 g, 43.3 mmol, 1 eq) in anhydrous DMF (25 mL) was stirred at room temperature under a No atmosphere for 72 h. The reaction mixture was diluted with H20 (500 mL) and extracted with Etoac (4 x 100 mL). The combined organic extracts were washed with H20 (100 mL) , satd NaCl(aq, (100 mL) , dried (MgS04) and evaporated in vacuo. The residue was triturated with n-hexane and filtered. The filtrate was evaporated in vacuo to give the product as a yellow oil which crystallised (11.02 g, 95%). 1H NMR (CDC13) 5 8.2 (d, J- 8.25 Hz, 2H), 7.47 (d, J - 8.02 Hz, 2H), 4.8 (s, 2H), 0.96 (s, 9H), 0.12 (s, 6H). IR (neat) 2955, 2930, 1605 and 1522 cin-l. HRMS m/z calcd for C13H22N03 Si 268.1369 (M + H) , found 268.13764- (tert-Butyl-dimethyl-silanyloxymethyl) -phenylamine (33) (Figure Removed) Ammonium formate was added to a solution of the nitro compound 32 (8.86 g, 33.1 mmol, 1 eq) in ethanol (175 mL) over 10% Palladium on carbon (2.66 g, 30 wt%) [caution exothermic] and the mixture was stirred at room temperature for 2.5 h. The reaction mixture was filtered through celite and the solvent evaporated in vacuo. The residue was partitioned between EtOAc (100 mL) and H20 (100 mL), the organic portion was washed with H20 (100 mL), satd NaCliaqi (100 mL), dried (MgSO 2-{3[4~ (tert-Butyl-dimethyl-silanyloxymethyl) -phenyl]-ureido}-pentanedioic acid diallyl ester (34) (Figure Removed) A solution of triethylamine (0.24 g, 0.33 mL, 2.4 mmol, 2 eq) in anhydrous CH2C12 (10 mL) was added dropwise to a solution of diallyl-L-glutamate tosylate (0.48 g, 1.2 mmol, leq) and triphosgene (0.12 g, 0.4 mmol, 0.3 eq) in anhydrous CH2C12 (40 mL) stirring at -80°C under a N2 atmosphere. The mixture was stirred at -80°C for 1 h then allowed to reach room temperature. A solution of the amino-silyl ether 33 (0.28 g, 1.2 mmol, 1 eq) and triethylamine (0.12 g, 0.17 mL, 1.2 mmol, 1 eq) in anhydrous CH2C12 (10 ml) was added dropwise (16 min) and the resulting solution stirred at room temperature for 18 h. The solvent was removed in vacua and the residue triturated with toluene and filtered. The filtrate was evaporated to give the crude product as a yellow oil. Purification by flash column chromatography (20% EtOAc/80% n-hexane) gave the product as a colourless oil (0.35 g, 61%). JH NMR (CDC13) 6 7.26 (s, 4H) , 6.83 (s, 1H) , 5.9 (m, 2H) , 5.61 (d, J = 8Hz, 1H) , 5.3 (m, 4H) , 4.68 (s, 2H) , 4.60 (m, 5H) , 2.5 (m, 2H) , 2.25 (m, 1H) , 2.) (m, 1H) , 0.95 (s, 9H) 0.07 (s, 6H) . IR (neat) 3349, 2954, 2929, 2856, 1739, 1650, 1603, 1553 cm' '. HRMS m/z calcd for C25H39N2O6 491 .2577 Si (M+H) , found 491.2554. 2- [3- (4-Hydroxymethyl-phenyl) ureido] -pentanedioic acid diallyl ester (35) A solution of the TBDMS ether 34 (0.32 g, 0.65 mmol) in AcOH/THF/HzO (9 mL/3 mL/3 mL) was stirred at room temperature for 2 h. The reaction mixture was cooled (ice bath) and neutralised with NaHC03(aq) (13.2 g, 15.7 mmol) in H2O (150 mL) . The mixture was extracted with EtOAc (4 x 30 mL) and the combined extracts were washed with H20 (100 mL) , satd NaCl(aq) (100 mL) , dried (MgS04) and evaporated in vacuo. Purification by flash column chromatography (60% EtOAc/40% n-hexane) gave the product as a colourless oil (0.22 g, 91%) ^H NMR (dfi DMSO) 6 8.5 (s, 1H) , 7.31 (d, J= 8.5Hz, 2H) , 7.16 (d, J «= 8.12 Hz, 2H) , 6.55 (d, J = S.lHz), 5.92 (m, 2H) , 5.3 (m, 4H) , 5.0 (t, J= 5.55HZ, 2H) , 4.6 (m, 4H) , 4.35 (m, 1H), 2.5 (m, 2H), 2.11 (m, 1H) , 1.9 (m, 1H) . IR (neat) 3353, 1738, 1659, 1602, 1551cm"1. HRMS m/z calcd for Ci9H25N206 377.1713 (M+H), found 377.1705. (Figure Removed)A solution of triethylamine (3.19 g, 4.4 mL, 31.6 mmol, 2.1 eq) in anhydrous CH2C12 (20 mL) was added dropwise to a solution of diallyl-L-glutamate tosylate (6.0 g, 15.0 mmol, leq) and triphosgene (1.6 g, 5.4 mmol, 0.36 eq) in anhydrous CH2Cl2 (40 mL) stirring at -80°C under a N2 atmosphere. The mixture was stirred at -80°C for 1.75 h then allowed to reach room temperature. A solution of hydroxyl-silyl ether 40, prepared using a literature procedure, (3.6 g, IS.Ommol 1 eq) and triethylamine (1.7 g, 2.3 mL, 16.5 mmol, 1.1 eq) in anhydrous CH2C12 (35 mL) was added dropwise and the resulting solution was stirred at room temperature for 18 h. The solvent was removed in vacuo and the residue triturated with toluene and filtered. The filtrate was evaporated to give the crude product as a yellow oil. Purification by flash column chromatography (20% EtOAc/80% n-hexane) gave the product as a colourless oil (3.6 g, 46%). *H NMR (CDC13) 5 7.3 (d, J= 8.5 Hz, 2H) , 7.08 (d, J= 8.5 Hz, 2H), 5.92 (m, 2H), 5.70 (d, J= 8 Hz, 1H) , 4.7 (s, 2H), 4.68 (d, J= 5.8 Hz, 2H), 4.60 (d, J= 5.8 Hz, 2H), 4.5 (m, 1H), 2.5 (m, 2H), 2.3 (m, 1H), 2.1 (m, 1H), 0.93 (s, 9H), 0.09 (s, 6H). IR (neat) 3350, 2954, 2930, 2865, 2857, 1738, 1531, 1503 cm"1. HRMS m/z calcd for C25H38N07 Si 492.2418 (M+H) , found 492.2411. 2~ (4-Hydroxymethyl-phenoxycarbony lamina) -pentanedioic acid diallyl ester (42) (Figure Removed) solution of the TBDMS ether 41 (3.23 g, 6.56 mmol) in AcOH/THF/H20 (18 mL/6 mL/6 mL) was stirred at room temperature for 2 h. The reaction mixture was cooled (ice bath) and neutralised with Na2C03(aq) (16.6 g, 15.6 mmol) in H20 (135 mL) . The mixture was extracted with EtOAc (4 x 150 mL) and the combined extracts were washed with H20 (250 mL) , satd NaCl(a (Figure Removed)A solution of vanillin (47)(40.00 g, 262.89 mmol) and methyl-4-bromobutyrate (50.00 g, 276.18 mmol) in DMF (200 ml) was allowed to stir over potassium carbonate (51.53 g, 372.40 iranol) for 16 hours. Water was added to the reaction mixture at which time the product crystallised. The resulting mixture was filtered and dried in vacuo for 16 hours to afford the keto-ester (48) as a white solid (41.3g, 66%). MP = 57-59°C. *H NMR (250 MHz, CDC13) 6 9.80 (s, 1H), 7.46-7.40 (m, 2H), 6.97 (d, J = 8.1 Hz, 1H), 4.16 (t, J- 6.3 Hz, 2H), 3.92 (s, 3H), 3.70 (s, 3H), 2.57 (t, J - 7.2 Hz, 2H) , 2.20 (pent, J= 6.7 Hz, 2H) . 13C NMR (67.8 MHz, CDC13) 188.2 (CD, 173.7 (C12) , 153.8 (Cguat.), 152.0 (Cquat.), 144.1 (Cquat.), 125.8 (Cmethine), 110.3 (C3), 108.5 (C6), 69.0 (C9), 57.0 (C8), 52.2 (C13), 30.6 (Cll), 24.5 (CIO). It was decided to adopt the numbering system shown in the figure below for the molecule for ease of peak assignment in 13C NMR. (Figure Removed)A solution of the keto-ester (48)(20.00 g, 79.3 mmol) in acetic anhydride (80 mL) was added dropwise to a stirring solution of HN03 (400 mL) and Ac20 (80 mL) at 0°C. After stirring for 2.5 hours the reaction mixture was poured into iced water (3L) and allowed to stand at 4°C for a further 16 hours. The precipitate was collected by filtration and dried under vacuum to provide the nitro compound (49) as a yellow crystalline solid (14.52 g, 62%). Melting Point 70-73°C. :H NMR (250 MHz, CDC13) 5 10.4 (s, 2H), 7.61 (s, 1H) , 7.40 (s, 1H), 4.21 (t, J= 6.2 Hz, 2H) , 4.00 (s, 3H), 3.71 (s, 3H), 2.58 (t, J=7.1Hz, 2H), 2.23 (pent, J=6.3 Hz, 2H) . 13C NMR (67.8 MHz, CDC12) 188.2 (Cl), 173.7 (C12) , 153.8 (Cquat.), 152.1 (Cquat.), 144.1 (Cquat.), 125.8 (Cquat.), 110.2 (C6), 108.5 (C3), 69.0 (C9) , 57.0 (C8), 52.2 (C13) , 30.6 (Cll), 24.4 (CIO). IR (cm'1) 3571, 3485, 2951, 1725, 1689, 1573, 1329, 1172, 1064, 1002, 936, 896, 858, 820, 766, 738, 688, 624. MS (M+l) 298.0. Anal. Calcd for CJ3 Hi5N07: C, 52.53; H, 5.09; N, 4.71. Found: C, 52.80; H, 5.03; N, 4.75. 4- (4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid methyl ester (50ium borohydride (2.69 g, 71.3 mmol) was added to a stirred solution of the ester (49)(10.0 g, 33.6 mmol) in THF (50 mL) under an N2 atmosphere at room temperature. Effervescence was observed upon addition of the reducing agent. TLC (EtOAc) after 16 hours revealed the complete loss of starting material. The solution was concentrated and redissolved in EtOAc (100 mL). The organic layer was washed with sat. NH 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid allyl ester (52) A mixture of compound 51 (7.00 g 24.50 mmol), allyl alcohol (80 mL) and p-tosic acid (742 mg, 3.90 mmol) was heated at reflux under a N2 atmosphere for 4 hOURS, at which time TLC (2% MeOH/CHCl3) indicated that reaction had gone to completion. Excess allyl alcohol was evaporated in vacuo to afford the crude compound (52), which was partitioned between NaHC03 (50 mL) and EtOAc (50 mL). The organic layer was washed with water (3 x 50 mL), brine (3 x 50 mL) and then dried (MgS04), and concentrated in vacua. Purification by flash chromatography (CHC13) yielded the product as an orange-brown solid (5.78 g, 72%). MP = 73-75°C JH NMR (250 MHz, CDC13) 5 7.70 (s, 1H), 7.16 (s, IE), 6.00-5.84 (m, 1H) , 5.36-5.21 (m, 2H) , 4.94 (d, J (pent, J= 6.7 Hz, 2H). 13C NMR 6173.1 (C12), 154.7 (Cquat.), 147.3 (Cquat.), 139.6 (Cquat.), 133.2 (Cquat.), 132.4 (C14), 118.7 (C15), 111.1 (C6), 109.9 (C3), 68.6 (C9), 65.6 (C13), 62.7 (Cl), 56.7 [2-12-(tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidine-J-carbonyl]-4,5-dimethoxy-phenyl]-carbamic acid-benzyl ester (9) solution of anhydrous benzyl alcohol (0.19 g, 0.18 mL, 1,1 iranol) and triethylamine (0.19 g, 0.26 mL, 1.85 mmol) in anhydrous toluene (5 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.522 g, 1.32 mmol), triphosgene (0.14 g, 0.48 mmol) and triethylamine (0.18 g, 0.25 mL, 1.8 mmol) in anhydrous toluene (15 mL) . The reaction was complete in 16 hours. The product was obtained (flash column chromatography 20% EtOAc/80% n-hexane) as a colourless oil (0.49 g, 70%): v^s/cnf1 (film) 1727 (C=0, carbamate) ; [ A solution of 2-trimethylsilyl ethanol (0.168 g, 0.2 mL, 1.42 mmol) and triethylamine (0.156 g, 0.22 mL, 1.55 mmol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.509 g, 1.29 mmol), triphosgene (0.138 g, 0.46 mmol) and triethylamine (0.176 g, 0.24 mL, 1.74 mmol) in anhydrous toluene (25 mL). The reaction was complete in 21 hours. The product was obtained (flash column chromatography 40%EtOAc/60% n-hexane) as a colourless oil (0.417 g, 60%): Vnax/cm"1 (film) (C=0, carbamate); ); [cc]D22-6- 96.15° (c = 0.21, CHC13), aH NMR (CDC13) 5 9.03 (bs, 1H) , 7.87 (s, 1H) , 7.87 (s, 1H), 6.82 (s, 1H), 4.35 (m, 1H) , 4.2 (m, 2H) , 4.01 (m, 1H) , , 3.50 (m, 2H) , 2.10 (m, 2H), 1.98 (m, 1H), 1.70 (m, 1H), 1.10 (m, 2H) , 0.9 (s, 9H), 0.08 (s, 15H), IR (neat) 3305, 2953, 2857, 1727, 1622, 1599, 1522 cm'1, [2-[2-(tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidine-1-caxbonyl]-4f5-dimethoxy-phenyl]-carbamic acid-3- (4-nitrophenyl)-allyl ester (12) lution of 3- (4-nitrophenyl)-prop-2-en-l-ol (0.222 g, 1.24 mmol) and triethylamine (0.138 g, 0.19 mL, 1.37 mmol) in anhydrous CH2Cl2 (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.491 g, 1.24 mmol), triphosgene (0.132 g, 0.45 mmol) and triethylamine (0.17 g, 0.23 mL, 1.68 mmol) in anhydrous toluene (20 mL). The reaction was complete in 20 hours. The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a yellow oil which crystallised (0.432g, 58%): v^/cm"1 (film) (C = 0, carbamate); [a]D25'3-65.0° (c = 0.24, CHC13), *H NMR (CDC13) 5 9.39 (bs, 1H) , 7.89 ( (s, 1H) , 8.19 (d, J= 8.8 Hz, 2H) , 7.53 (d, J= 9 Hz, 2H) , 6.85 (s, 1H), 6.73 (d, J= 15.9 Hz, 1H), 6.52 (dt, J= 5.8, 15.9 Hz, 1H) 4.86 (m, 2H) 4.38 (m, 1H), 4.01 (m, 1H), 3.94 (s, 3H), 3.83 (s, 3H), 3.70 (m, 1H), 3.52 (m, 2H), 2.06 (m, 2H), 1.95 (m, 1H), 1.72 (m, 1H), 0.9 (s, 9H), 0.08 (s, 6H). IR (neat) 3340, 2930, 1729, 1597, 1519 cm"1. HRMS m/z calcd for CaoHaiNsOeSiNa 622.2561 (M+Na) , found 622.2542. {2- [-(tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidine-1-carbonyl]-4,5-dimethoxy-phenyl)-carbamic acid 2-benzenesulfonyl-ethyl ester (13)A solution of phenylsulfonylethanol (0.26 g, 0.167 mL 1.39 mmol) and triethylamine (0.13 g, 0.18 mL, 1.29 mmol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.5 g, 1.27 mmol), triphosgene (0.135 g, 0.34 mmol) and triethylamine (0.17 g, 0.24 mL, 1.72 mmol) in anhydrous toluene (20 mL) . The reaction was complete in 96 hours. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a yellow oil (0.3 g, 39%) . [cc]D 24'a - 81.5° (c = 0.23, CHC13), *H NMR (CDC13) 6 9.1 (s, 1H) , 7.95 (d, J = 8 Hz, 2H), 7.72 (s, 1H), 7.58 (m, 3H), 6.82 (s, 1H), 4.45 (t, J= 6.3 Hz, 2H) , 4.35 (m, 1H) , 3.95 (m, 1H) , 3.91 (s, 3H) , 3.82 (s, 3H), 3.70 (m, 1H), 3.5 (m, 4H), 2.1 (m, 2H), 1.95 (m, 1H), 1.7 (m, 1H), 0.9 (s, 9H), 0.05 {s, 6H). Acetic acid l—[2- (4,5-dimethoxy-2-nitro-benzyloxycarbonylamino)-4,5-dimethoxy-benzoyl]-pyrrolidin-2-ylmethyl ester solution of nitroveratryl alcohol (1.98 g, 9.3 mmol) and triethylamine (0.94 g, 1.29 mL, 9.3 mmol) in anhydrous CH2C12 (30 mL) was added to a solution of the isocyanate 8 prepared from the amine 6 (3.0 g, 9.3 mmol), triphosgene (0.99 g, 3.4 mmol) and triethylamine (1.27 g, 1.75 mL, 12.6 mmol) in anhydrous toluene (60 ml>) . The reaction was complete in 16 h. The product was obtained (flash column chromatography 40% EtOAc/60% n-hexane) as a yellow foam (4.15 g, 69%). [a]"D -62.5° (c = 1.0, CHC13) ; JH NMR (CDC13) 5 9.16 (s, 1H) , 7.80 (s, 1H) , 7.74 (s, 1H) , 7.10 (s, 1H), 6.83 (s, 1H), 5.65 (d, J= 15.2 Hz, 1H) , 5.55 (d, J= 15.2 Hz, 1H), 4.58 (bs, 1H) , 4.29 (bs, 2H) , 3.99 - 3.86 (m, 12H) , 3.66 - 3.46 (m, 2H) , 2.07 (s, 3H) , 2.03 - 1.90 (m, 1H) , 1.90 - 1.67 (m, 3H) ; 13C' NMR (CDC13) 5 169.5, 153.7, 153.2, 151.3, 148.1, 143.9, 139.6, 132.2, 127.9, 115.3, 111.2, 109.9, 108.2, 104.4, 64.4, 63.6, 56.5, 56.4, 56.4, 56.0, 55.9, 50.8, 27.7, 25.0, 20.8; MS (AP) m/z (relative intensity) 584 (M+Na) , 562 (M+l) ; IR (neat) 3333, 2939, 1735, 1672, 1600, 1520, 1462, 1395, 1327, 1274, 1221, 1173, 1114, 1072, 1036, 873, 795.1, 756.1 (cm'1); HRMS m/z calcd for C26H32N3OU 562.2033 (M+H) found 562.2037. 4- {4- [2- (2-Acetoxymethyl-pyrrolidine-l-carbonyl) -4 ,5-phenylcarba^loyloxymethylJ-2-Inethoxy-5-nitro-phenoxy}butyric acid allyl ester (15) A solution of linker alcohol (52) (3.02 g, 9.3 mmol) and triethylamine (0.94 g, 1.29 mL, 9.3 mmol) in anhydrous CH2Cl2/toluene (20 mL/10 mL) was added to a solution of the isocyanate 8 prepared from the amine 6 (3.0 g, 9.3 mmol), triphosgene (0.99 g, 3.4 mmol) and triethylamine (1.27 g, 1.75 mL, 12.6 mmol) in anhydrous toluene (100 mL). The reaction was complete in 16 hours. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a yellow foam (1.84 g, 30%). [a]asD -54.5° (c - 1.0, CHC13) ; 1H NMR (CDC13) 6 9.15 (s, 1H) , 7.80 (s, 1H), 7.73 (s, 1H), 7.09 (s, 1H), 6.83 (s, 1H), 5.98 -5.87 (m, 1H), 5.69 (d, J= 15.2 Hz, 1H), 5.53 (d, J= 15.2 Hz, 1H) , 5.35 - 5.22 (m, 2H), 4.62 - 4.58 (m, 3H), 4.29 (bs, 2H) , 4.18 - 4.11 (m, 2H), 3.97 (s, 3H), 3.92 (s, 3H), 3.89 (s, 3H) , 3.58 - 3.51 (m, 2H), 2.59 (t, J= 7.2 Hz, 2H), 2.24 - 2.15 (m, 2H), 2.06 (s, 3H), 1.98 - 1.79 (m, 4H) ; 13C NMR (CDC13) 6 172.5, 170.9, 169.4, 154.1, 153.2, 151.2, 147.3, 143.9, 139.5, 132.1, 128.0, 118.4, 115.3, 111.2, 110.1, 109.6, 104.4, 68.2, 65.3, 64.5, 63.7, 56.6, 56.4, 56.0, 55.9, 50.9, 30.6, 27.6, 25.0, 24.2, 20.9; MS (AP) m/z (relative intensity) 673 (M+', 15), 515 (14), 487 (4), 400 (3), 349 (9), 322 (22), 308 (30), 289 (8), 247 (4), 224 (6); IR (neat) 2940, 1735, 1600, 1520, 1465, 1395, 1326, 1275, 1228, 1173, 1114, 1069, 1037, 993, 939, 871, 755 cm"1; HRMS m/z calcd for C32H39N3Oi3 673.2494 (M+) found 673.2483. 2-[3- (4~{2~[2~ (tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidine-1 -carbonyl] -4,5-dimethoxy-phenylcarbamoyloxymethyl} -phenyl) -ureido]-pentanedioic acid diallyl ester (36) A solution of the urea progroup 35 (2.03 g, 5.4 mmol) and triethylamine (0.6 g, 0.83 mL, 5.95 iranol) in anhydrous CH2C12 (55 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (2.13 g, 5.4 iranol), triphosgene (0.58 g, 1.95 iranol) and triethylamine (0.74 g, 1.01 mL, 7.3 mmol) in anhydrous toluene (70 mL). The reaction was complete in 20 h. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a white foam (1.94 g, 45%). [cc]D24-8- 40.6* (c - 0.234, CHC13), 1H NMR (CDC13) 6 9.1 (bs, 1H), (7.85 (5, 1H), 7.31 (s, 1H), 6.87 (s, 1H), 6.82 (s, 1H), 5.90 (m, 2H), 5.62 (d, J= 11.7 Hz, 1H), 5.25 (m, 4H), 5.11 (s, 2H), 4.63 (m, 5H), 4.35 (m, 1H), 3.99 (m, 1H), 3.92 (s, 3H) , 3.81 (s, 3H), 3.63 (m, 1H), 3.51 (m, 2H), 2.50 (m, 2H), 2.25 (m, 1H), 2.00 (m, 3H) , 1.70 (m, 1H), 0.9 (s, 9H), 0.05 (s, 6H) . IR (neat) 3349, 2952, 2856, 1732, 1664, 1600, 1520 cm'1. HRMS w/z calcd for C^H^N/AiSlCs 929.2769 (M+Cs) , found 929.2727. 2- (4- {2- [2- (tert-Butyl-dimethyl-silanyloxymethyl) -pyrrolidine-1-carbonyl ] -4 , 5-dimethoxy-phenylcarbamoyloxymethyl} -phenoxycarbonylamino) -pentanedioic acid diallyl ester (43) MeO 0 A solution of the carbamate progroup 42 (1.48 g, 3.92 iranol) and triethylamine (0.45 g, 0.6 mL, 4.3 mmol) in anhydrous CH2C12 (30 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (1.54 g, 3.92 inmol)/ triphosgene (0.42 g, 1.4 iranol) and triethylamine (0.535 g, 0.74 mL, 5.3 mmol) in anhydrous toluene (50 mL). The reaction was complete in 4 h. The product was obtained (flash column chromatography 40% EtOAc/60% n-hexane) as a colourless oil (1.76 g, 56). aH NMR (CDC13) 6 9.22 (bs, 1H), 7.87 (s, 1H) , 7.40 (d, J = 10 Hz, 2H), 7.13 (d, J= 7.5 Hz, 2H), 6.83 (s, 1H), 5.93 (m, 2H), 5.74 (d, J= 8.3 Hz, 1H), 5.30 (m, 4H) , 5.17 (d, J- 12.5 Hz, 1H), 5.15 (d, J= 12.5 Hz, 1H) 4.68 (d, J- 5 Hz, 2H) 4.61(d, J - 5 Hz, 1H), 4.48 (m, 1H), 4.35 (m, 1H), 3.95 (s, 3H), 3.82 (s, 3H), 3.65 (m, 1H), 3.50 (m, 2H), 2.55 (m, 2H), 2.30 (m, 1H), 2.1 (m, 3H), 1.95 (m, 1H), 1.8 (m, 1H), 0.9 (s, 9H), 0.04 (s, 6H). IR (neat) 3338, 2953, 2857, 1735, 1615, 1597, 1522 cm"1. {2-[2-tert-Butyl-dimethyl-silanyloxymethyl]-pyrrolidine-l-carbonyl}-4,5-dimethoxy-phenyl}-carbamic acid methyl ester (77) O A solution of methanol (0.39 g, 0.5 mL 12.3 mmol) and triethylamine (0.3 g, 0.41 mL, 2.95 mmol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.97 g, 2.5 mmol), triphosgene (0.263 g, 0.89 mmol) and triethylamine (0.335 g, 0.46 mL, 3.32 romol) in anhydrous toluene (30 mL) . The reaction was complete in 18 hours. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a colourless oil (0.694 g, 62.5%): [a]D 2S -122" (c = 0.24, CHC13) ; XH NMR (CDC13) 6 9.1 (bs, 1H) , 7.86 (s, 1H) , 6.83 (s, 1H), 4.35 (bs, 1H), 4.05 (m, 1H), 3.93 (s, 3H), 3.82 (s, 3H), 3.74 (s, 3H) , 3.67 (m, 1H) , 3.5 (m, 2H) , 2.07 (m, 2H) , 1.95 (m, 1H) 1.72 (m, 1H) , 0.90 (s, 9H) , 0.04 (s, 6H) ; MS (ES+) m/z (relative intensity) 475.2 (M+- +Na, 5), 453.2 (M*- +1, 100); IR (neat) 2953, 1733, 1598, 1524, 1396 cm"1. (2-[2-tert-Butyl-dimethyl-silanyloxymethyl] -pyrrolidine-1-carbonyl}-4,5-dimethoxy-phenyl}-carbamic acid tert-butyl ester (78) A solution of t-butanol (0.99 g, 1.25 mL 13.3 mmol) and triethylamine (0.292 g, 0.4 mL, 2.9 mmol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (1.05 g, 2.66 mmol), triphosgene (0.284 g, 0.96 mmol) and triethylamine (0.36 g, 0,5 mL, 3.6 mmol) in anhydrous toluene (30 mL). The reaction was stirred at room temperature for 18 hours then heated under reflux for 18 h. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a colourless oil (0.392 g, 30%): [ot]D23-5- 90.7° (c= 0.193, CHC13) ; :H NMR (CDC13) 5 8.58 (bs, 1H) , 7.91 (s, 1H), 6.82 (s, 1H), 4.37 (bs, 1H), 4.13 (m, 1H), 3.95 (s, 3H) , 3.82 (s, 3H), 3.68 (m, 1H), 3.51 (m, 2H), 2.05 (m, 2H), 1.93 (m, 1H), 1.74 (m, 1H), 1.61 (s, 9H), 0.97 (s, 9H), 0.04 (s, 6H); MS (ES+) m/z (relative intensity) 517.2 (M+- +Na, 5), 495.2 (M+- +1, 100); IR (neat) 2930, 1723, 1600, 1521, 1420, 1394 cm'1. {2~[2-tert-Butyl-dimethyl-silanyloxymethyl]-pyrrolidine-l-carbonyl}-4,5-dimethoxy-phenyl}-carbamic acid 2,2,2-trichloroethyl ester (79)A solution of 2,2,2-trichloroethanol (0.48 g, 0.31 mL 3.2 mmol) and triethylamine (0.36 g, 0.49 mL, 3.5 mmol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (1.15 g, 2.92 mmol), triphosgene (0.31 g, 1.05 mmol) and triethylamine (0.398 g, 0.55 mL, 3.94 mmol) in anhydrous toluene (30 mL). The reaction was complete in 18 h. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a colourless oil (1.075 g, 65%): [a]D 24>z- 90.5* (c = 0.21, CHC13); *H NMR (CDC13) 5 9.48 (bs, 1H) , 7.85 (s, 1H) , 6.87 (s, 1H) , 4.83 (d, J= 11.97 Hz, 1H) , 4.76 (d, J= 12.0 Hz, 1H) , 4.39 (bs, 1H), 3.99 (m, 1H), 3.94 (s, 3H), 3.84 (s, 3H), 3.68 (m, 1H), 3.56 (m, 2H), 2.1 (m, 2H) , 1.98 (m, 1H), 1.74 (m, 1H), 0.91 (s, 9H), 0.04 (s, 6H) ; MS (ES+) m/z (relative intensity) 593.0 (M+- +Na, 5), 571.1 (M+> +1, 100); IR (neat) 2953, 1746, 1599, 1524, 1462, 1422, 1397 cm'1.{2-[2-tert-Butyl-dimethyl-silanyloxymethyl]-pyrrolidine-1-carbonyl} -4,5-diinethoxy-phenyl}-carbamic acid 4-nitro benzyl ester (80) mine (0.3 g, 0.42 mL, 3.0 mmol) in anhydrous dichloromethane (20 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.985 g, 2.5 mmol), triphosgene (0.265 g, 0.895 mmol) and triethylamine (0.34 g, 0.47 mL, 3.36 mmol) in anhydrous toluene (40 mL). The reaction was complete in 18 h. The product was obtained (flash column chromatography 40% EtOAc/60% n-hexane) as a yellow oil (0.87 g, 61%): [a]D21-°- 85.15° (c = 0.23, CHC13) ; *H NMR (CDC13) 69.48 (bs, 1H) , 8.22 (d, J = 8.8 Hz, 2H), 7.87 (s, 1H), 7.56 (d, J= 8.8 Hz, 2H), 6.86 (s, 1H), 5.26 (2d, J = 13.5 Hz, 2H), 4.36 (bs, 1H), 4.0 (m, 1H), 3.93 (s, 3H), 3.83 (s, 3H), 3.67 (m, 1H), 3,55 (m, 2H), 2.06 (m, 2H) , 1.97 (m, 1H), 1.75 (m, 1H), 0.90 (s, 9H), 0.03 (s, 6H); MS (ES+) m/z (relative intensity) 482.2 (M*- +Na, 100), 460.3 (M+- +1, 60); IR (neat) 2953, 1728, 1600, 1519, 1397, 1346 cm'1. {2-[2-tert-Butyl-dimethyl-silanyloxymethyl] -pyrrolidine-1-carbonyl}-4,5-dimethoxy-phenyl}-carbamic acid 9H-fluorenyl-9-ylmethyl ester (81) A solution of 9-fluorenylmethanol (0.54 g, 2.7 nunol) and triethylamine (0.3 g, 0.4 mL, 3.0 iranol) in anhydrous toluene (20 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (0.98 g, 2.5 mmol) , triphosgene (0.265 g, 0.895 nunol) and triethylamine (0.34 g, 0.47 mL, 3.36 mmol) in anhydrous toluene (40 mL) . The reaction was complete in 18 h. The product was obtained (flash column chromatography 20% EtOAc/80% n-hexane) as a yellow oil (1.04 g, 68%): [a]D20-8- 72.96° (c = 0.23, CHC13) ; 1H NMR (CDC13) 6 9.32 (bs, 1H) , 7.86 (s, 1H) , 7.78 (d, J= 7.5 Hz, 2H) , 7.66 (m, 2H) , 7.42 (d, J= 7.4 Hz, 2H) , 7.34 (d, J = 7.4 Hz, 2H), 6.9 (s, 1H), 4.51-4.28 (m, 4H) , 4.05 (m, 1H) , 3.94 (s, 3H) , 3.85 (s, 3H), 3.72 (m, 1H) , 3.55 (m, 2H) , 2.09 (m, 2H) , 1.98 (m, 1H) , 1.75 (m, 1H), 0.93 (s, 9H) , 0.07 (s, 6H) ; MS (ES4) m/z (relative intensity) 525.2 (M+- 4Na, 100), 503.3 (M+- +1, 55); IR (neat) 2953, 1727, 1600, 1522, 1397, 1346 cnf1. (2- [2-tert-Butyl-dimethyl-silanyloxymethyl] -pyrrol idine-1 -carbonyl}-4,5-dimethoxy-phenyl}-carbamic acid allyl ester (82) A solution of allyl alcohol (3.73 g, 4.36 mL, 64.15 mmol) and triethylamine (1.56 g, 2.14 mL, 15.4 mmol) in anhydrous toluene (40 mL) was added to a solution of the isocyanate 7 prepared from the amine 5 (5.06 g, 12.8 mmol), triphosgene (1.37 g, 4.62 mmol) and triethylamine (1.75 g, 2.41 mL, 17.3 mmol) in anhydrous toluene (120 mL) . The reaction was complete in 48 h. The product was obtained (flash column chromatography 30% EtOAc/70% n-hexane) as a pale yellow oil (4.23 g, 69%): [cc]D20-2- 106.7* (c = 0.45, CHC13); JH NMR (CDC13) 6 9.16 (bs, 1H) , 7.88 (s, 1H) , 6.84 (s, 1H) , 6.00-5.93 (m, 1H) , 5.38-5.23 (m, 2H) , 4.66-4.63 (m, 2H) , 4.36 (bs, 1H), 4.0 (m, 1H) , 3.94 (3, 3H) , 3.83 (s, 3H) , 3.7 (m, 1H) , 3.52 (m, 2H), 2.08-2.04 (m, 2H) , 1.97 (m, 1H) , 1.75 (m, 1H) , 0.91 (s, 9H), 0.04 (s, 6H) / MS (ES+) m/z, (relative intensity) 501.1 (M4' +Na, 3), 479.1 (M*' +1, 100); Ir (neat) 2953, 2857, 1731, 1622, 1599, 1524, 1397 can"1 Deprotection of alcohols General methods for the deprotection of monomer tert- butyldimethylsilyl ethers and acetates Method A (TBDMS ethers) A 1.0 M THF solution of tetra-N-butyl-ammonium fluoride (1.2 eq.) was added via syringe to a solution of the TBDMS ether (1 eq.) in THF at 0°C. The reaction was stirred at room temperature until reaction was complete (TLC). The solvent was removed in vacuo and the product purified by flash column chromatography. Method B (TBDMS ethers) A solution of the TBDMS ether in a mixture of AcOH/THF/H20 (3/1/1) was stirred at room temperature until reaction was complete (TLC). The reaction mixture was cooled (ice bath) and carefully neutralised with KaHC03 (aq) (1 eq) . The mixture was extracted with EtOAc (x3) the combined extracts were washed with water (xl), satd NaCl (aq) (xl), dried (MgSO«) and evaporated in vacuo. The product was purified by flash column chromatography. Method C (acetates) A solution of K2C03 (5 eq) in H2O was added dropwise to a solution of the acetate (1 eq) in MeOH/CHCl3. The mixture was stirred at room temperature until reaction was complete (TLC). The solvent was evaporated in vacuo and the aqueous portion washed with EtOAc (x3) . The combined organic extracts were washed with satd NaCl (aq) (x3) , dried (MgSO e TBDMS ether 9 (0.517 g, 0.98 mmol) in THF (15 mL) was deprotected (Method A: Bu4NF (1.2 mL, 1.2 mmol)) to give the product (flash column chromatography 90% EtOAc/10% n-hexane) as a colourless oil (0.3 g, 75%). ) ; [ [2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenyl]-carbamic scid-4-methoxy-benzyl ester (17) MeO MeO, The TBDMS ether 10 (0.37 g, 0.66 mmol) in THF (20 mL) was deprotected (Method A: Bu^NF (0.8 ml, 0.8 mmol)) to give the product (flash column chromatography 80% EtOAc/20% n-hexane, then EtOAc) as a colourless oil (0.29 g, 97%). [a]D22'1- 69.6° (c = 0.23, CHC13), *H NMR (CDC13) 6 8.72 (s, 1H) , 7.76 (s, 1H) , 7.34 (d, J= 8.6 Hz, 2H) , 6./90 (d, J «= 8.6 Hz, 2H) , 6.80 (s, 1H) , 5.10 (m, 2H), 4.40 (m, 1H), 4.30 (m, 1H), 3.92 (s, 3H), 3.83 (s, 3H), 3.80 (m, 4H), 3.7 (m, 1H), 3.58 (m, 1H), 3.45 (m, 1H), 2.15 (m, 1H), 1.90 (m, 1H), 1.70 (m, 2H). IR (neat) 3338, 2958, 2837, 1726, 1599 cm'1. 68 [2- (2-Hydroxymethyl-pyrrolidine~l-carbonyl) -4,5-dimethoxy-phenyl]~carbamic acid 2-trimethylsilanyl-ethyl ester (18) The TBDMS ether 11 (0.354 g, 0.66 iranol) was deprotected (Method B: AcOH (6 mL)/THF (2 mL)/H20 (2 mL) then NaHC03 (8.77 g, 104.4 nunol) in H20 (100 mL)) to give the product (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless oil (0.27 g, 95%) .%) . [cx]D 22'° - 91.0* (c = 0.21, CHC13) , *H NMR (CDC13) 8 8.62 (s, 1H), 7.8 0 (s, 1H) , 6.81 (s, 1H) , 4.42 (m, 1H) , 4.30 (m, 1H) , 4.22 (m, 2H), 3.93 (s, 3H), 3.85 (m, 4H) , 3.70 (m, 1H), 3.60 (m, 1H), 3.50 (m, 1HO, 2.20 (m, 1H), 1.90 (m, 1H) 1.70 (m, 2H), 1.09 (m, 2H), 0.08 (s, 9H). IR (neat) 3340, 2953, 1726, 1597, 1520 cm"1. [2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl) -4,5-dimethoxy-phenyl]-caibamic acid-3-(4-nitrophenyl)-allylester (19) The TBDMS ether 12 (0.346 g, 0.58 mmol) was deprotected (Method B: AcOH (6 mL)/THF (2 mL)/H2O (2 mL) then NaHC03 (8.77 g, 104.4 mmol) in H20 (100 mL)) to give the product (flash column chromatography EtOAc) as a yellow foam (0.26 g, 92%) . [a]D 21'9-61.2* (c = 0.24, CHC13), *H NMR (CDC13) 68.95 {s, 1H) , 8.20 (d, J = 8.8 Hz, 2H), 7.81 (s, 1H), 7.53 (d, J- 8.8 Hz, 2H), 6.84 (s, 1H), 6.75 (d, J = 16 Hz, 1H), 6.50 (dt, J- 5.8, 16 Hz, 1H), 4.84 m, 2H), 4.43 (m, 1H), 4.25 (m, 1H) , 3.94 (s, 3H) , 3.86 (m, 4H) , 3.75 (m, 1H), 3.65 (m, 1H) , 3.55 (m, 1H) , 2.70 (m, 2H) , 2.20 (m, 1H), 1.90 (m, 1H). IR (neat) 3340, 2939, 1728, 1597, 1519 cm"1. [2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl) -4,5-dimethoxy-phenylj-carbamic acid 2-benzenesulfonyl-ethyl ester (20)The TBDMS ether 13 (0.3 g, 0.49 mmol) was deprotected (Method B: AcOH (9 mL)/THF (6 mL)/H20 (3 mL) then NaHCO3 (13.16 g, 156.6 mmol) in H20 (175 mL)) to give the product (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless oil (0.13 g, 54%) . [a]D 22'° - 50.7° (c = 0.22, CHC13) , *H NMR (CDC13) 6 8.79 (s, 1H), 7.80 (s, 1H), 7.40 (m, 4H) , 6.82 (s, 1H), 5.20 (d, J = 12.3 Hz, 1H), 5.16 (d, J= 12.3 Hz, 1H), 4.40 (m, 1H), 4.25 (m, 1H), 3.93 (s, 3H), 3.85 (m, 4H), 3.70 (m, 1H), 3.60 (m, 1H), 3.50 (m, 3H), 2.20 (m, 1H), 1.90 (m, 1H), 1.70 (m, 2H). IR (neat) 3337, 2938, 1729, 1598, 1524cm-1.[2- (2-Hydroxymethyl-pyrrolidine-1 -carbonyl) -4,5-dimethoxy-phenylj-carbamic ac±d 4f5-dimethoxy-2-nitro-benzyl ester (21) The acetate 14 (2.0 g, 3.5 mmol) in MeOH/CHCl3 (80 mL/30 mL) was deprotected (Method C: K2C03 (2.45 g, 18.0 mmol) in H2O (50 mL)) to give the product (flash column chromatography 2% MeOH/98% CHC13) as a yellow foam (1.6 g, 85D -59.0° (c = 1.0, CHC13) ; :H NMR (CDC13) 5 8.99 (s , 1H) , 7.72 (s, 2H), 7.10 (s, 1H) , 6.83 (s, 1H) , 5.63 (d, J= 15.2 Hz, 1H) , 5.52 (d, J= 15.2 Hz, 1H) , 4.38 (bs, 2H) , 4.08 - 3.80 (m, 13H) , 3.73 - 3.44 (m, 2H) , 2.25 - 2.08 (m, 1H) , 2.03 - 1.60 (m, 3H) ; 13C NMR (CDC13) 6 170.6, 153.7, 153.1, 151.1, 148.1, 144.1, 139.6, 131.4, 127.7, 110.7, 110.0, 108.2, 104.6, 66.0, 63.8, 60.8, 56.5, 56.4, 56.0, 51.4, 28.2, 25.0; MS (FAB) jn/z 652 (M+Cs), 520 (M+l); IR (neat) 3362, 2960, 1735, 1613, 1519, 1454, 1397, 1321, 1279, 1220, 1174, 1119, 1072, 1034, 990, 958, 880, 847, 795, 758, 708 cm"1; HRMS m/z calcd for 20.1946 (M+H) found 520.1931. 4-{4-[2- (2-hydroxymethyl-pyrrolidine-l-carbonyl) -4,5-dimethoxy-phenylcarbamoyloxymethyl] -2-methoxy-5-nitro-phenoxy)-butyric acid methyl ester (22) The acetate 15 (1.80 g, 2.7 mmol) in MeOH/CHCl3 (80 mL/30 mL) was deprotected (Method C: K2C03 (1.83 g, 13.3 mmol) in H20 (50 mL) ) to give the methyl ester product as a yellow foam (1.06 g, 63%). [a]24D -50.0° (c = 1.0, CHC13) ; 1E NMR (CDC13) 6 8.98 (s, 1H) , 7.72 (s, 2H), 7.08 (s, 1H), 6.83 (s, 1H) , 5.63 (d, J - 15.2 Hz, 1H) , 5.52 (d, J- 15.0 Hz, 1H), 4.41 - 4.31 (m, 1H) , 4.18 - 4.09 (m, 3H), 3.97 (s, 3H), 3.91 (s, 3H) , 3.85 (s, 3H), 3.66 (s, 3H), 3.62 - 3.50 (m, 2H), 2.59 - 2.54 (m, 2H) , 2.19 (pent., J = 6.7 Hz, 2H), 1.91 - 1.66 (m, 4H); 13C NMR (CDC13) 6 174.3, 171.7, 154.9, 154.1, 151.9, 148.2, 144.8, 140.2, 132.2, 128.5, 119.0, 111.3, 110.8, 110.1, 105.2, 68.3, 66.2, 63.8, 60.9, 56.5, 56.4, 56.0, 51.7, 51.5, 30.3, 28.3, 25.1, 24.2; MS (FAB) m/z 628 (M+Na) 606 (M+l); IR (neat) 3342, 2953, 2615, 1740, 1601, 1531, 1398, 1333, 1285, 1116, 1072, 996, 943, 870, 814, 755 cm'1. The TBDMS ether 36 (1.88g, 2.36 mmol) was deprotected (Method B: AcOH (18 mL)/THF (6 mL) /H20 (6 mL) then NaHC03 (26.3 g, 313.0 mmol) in H20 (270 mL)) to give the product (flash column chromatography 99% EtOAc/MeOH 1%) as a white foam (1.6 g, 99 %). [a],,"'*- 45.9' (c = 0.21, CHC13) , :H NMR (CDC13) 5 8.69 (s, 1H) , 7.75 (s, 1H), 7.30 (m, 4H) , 7.12 (s, 1H), 6.80 (s, 1H), 5.90 (m, 2H) , 5.74 (d, J «= 12.8 Hz, 1H) , 5.30 (m, 4H) , 5.10 (s, 2H) , 4.60 (s, 5H), 4.40 (ra, 1H), 4.20 (m, 1H), 3.90 (m, s, 3H), 3.84 (m, 4H), 3.70 (m, 1H), 3.50 (m, 2H), 2.50 (m, 2H), 2.10 (m, 2H), 1.90 (m, 1H), 1.70 (m, 2H). IR (neat) 3350, 2951, 2856, 1734, 1660, 1601, 1518 cm"1. HRMS m/z calcd for C39HoN4OnNa 705.2748 (M+Na) , found 705.2721. 2-(4-{2-[2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenylcarbamoyloxymethyl] -phenoxycarbonylamino} -pentanedioic acid diallyl ester (44) e TBDMS ether 43 (2.17 g, 2.72 mmol) was deprotected (Method B: AcOH (21 mL)/THF (7 mL)/H20 (7 mL) then NaHC03 (30.66 g, 365.0 mmol) in H20 (400 mL)) to give th chromatography EtOAc) as a white foam (1.7 g, 91 %). *H NMR (CDC13) 5 8.77 (s, 1H), 7.68 (s, 1H), 7.42 (d, J= 7.5 Hz, 2H), 7.13 (d, J= 7.5 Hz, 2H), 6.79 (s, 1H), 5.95 (m, 2H), 5.78 (d, J= 9.4 Hz, 1H), 4.70 (m, 4H), 4.50 (m, 1H), 4.40 (m, 1H), 4.25 (m, 1H), 3.93 (s, 3H), 3.84 (m, 4H), 3.70 (m, 1H), 3.55 (m, 2H), 2.55 (m, 2H) 2.40 (m, 1H), 2.10 (m, 2H), 1.90 (m, 1H), 1.70 (m, 2H) . IR (neat) 3345, 2950, 2856, 1736, 1615, 1598, 1522 cm'1. (2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenyl]-carbamic acid methyl ester (83) The TBDMS ether 77 (0.67 g, 1.5 mmol) in THF (30 mL) was deprotected (Method A: Bu4NF (1.74 mL, 1.74 mmol)) to give the product (flash column chromatography 99% EtOAc/1% MeOH) as a colourless oil (0.46 g, 99%): [a]D25-5- 125.0* (c = 0.22, CHC13) ; JH NMR (CDC13) 6 8.72 (bs, 1H) , 7.76 (s, 1H) , 6.81 (s, 1H), 4.42- 4.14 (m, 2H), 3.92 (s, 3H), 3.84 (s + m, 4H), 3.6-3.4 (m, 2H), 2.17 (m, 1H), 1.90-1.62 (m, 3H); MS (ES+) m/z (relative intensity) 361.0 (M4- -fNa, 20), 339.1 (M*' +1, 100); IR (neat) 3339, 2953, 1730, 1598, 1524, 1458, 1397 cm"1. [2-(2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenyl]-carbamic acid tert-butyl ester (84) he TBDMS ether 78 (0.34 g, 0.69 mmol) in THF (20 mL) was deprotected (Method A: Bu4NF (0.83 mL, 0.83 mmol)) to give the product (flash column chromatography EtOAc) as a colourless oil (0.26 g, 99%): [a]D24>6- 90.4* (c = 0.19, CHC13) ; aH NMR (CDC13) 5 8.42 (bs, 1H), 7,81 (s, 1H), 6.80 (s, 1H), 4.43 (m, 1H), 4.28 (bs, 1H), 3.94 (s, 3H), 3.84 (s -f m, 4H) , 3.76 (m, 1H) , 3.62 (m, 1H), 3.51 (m, 1H), 2.17 (m, 1H), 1.91 (m, 1H) 1.85-1.62 (m, 2H), 1.51 (s, 9H); MS (ES+) m/z (relative intensity) 403.1 (M+' +Na, 15), 381.1 (M4" + 1, 100); IR (neat) 3340, 2975, 1721, 1597, 1522, 1456, 1395 cm"1. [2-(2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenyl]-carbamic acid 2,2,2-trichloro-ethyl ester (85) (Figure Removed)The TBDMS ether 79 (1.03 g, 1.8 nunol) was deprotected (Method B: AcOH (15 roL)/THF (5 raL)/H20 (5 mL) then NaHC03 (21.9 g, 261 mmol) in H2O (300 mL)) to give the product (crystallised from 80% EtOAc/20% n-hexane) as a white solid (0.8 g, 97%): [a]D24'3- 91-4" (c = 0.19, CHC13) ; JH NMR (CDC13) 6 9.04 (bs, 1H) , 7.73 (s, 1H) , 6.85 (s, 1H), 4.83 (d, J= 12.0 Hz, 1H), 4.78 (d, J= 12.0 Hz, 1H>, 4.44 (m, 1H), 4.05 (m, 1H) , 3.93 (s, 3H) , 3.86 (s + m, 4H), 3.73 (m, 1H), 3.61-3.5 (m, 2H), 2.04 (m, 1H), 1.92 (m, 1H), 1.73 (m, 2H); MS (ES+) m/z (relative intensity) 477.0 (M+~ +Na, 30), 455.0 (M+> +1, 100); IR (neat) 3306, 2954, 1743, 1599, 1524, 1432, 1396 cm"1. [2-(2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy-phenylj-carbamic acid 4-nitro-benzyl ester (86) (Figure Removed) The TBDMS ether 80 (0.81 g, 1.4 mmol) was deprotected (Method B: AcOH (15 mL)/THF (5 mL)/H20 (5 mL) then NaHCO3 (21.9 g, 261 mmol) (Figure Removed) in H20 (300 mL)) to give the product (flash column chromatography EtOAc) as a pale yellow oil (0.61 g, 94%): [a]D22-°- 60.42° (c = 0.24, CHC13); :H NMR (CDC13) 6 9.05 (bs, 1H) , 8.23 (d, J= 8.8 Hz, 2H), 7.78 (s, 1H), 7.56 (d, J= 8.8 Hz, 2H) , 6.84 (s, 1H) , 5.27 (m, 2H), 4.42 (m, 1H), 4.15 (m, 1H) , 3.93 (s, 3H), 3.85 (s + m, 4H), 3.71 (m, 1H), 3.62 (m, 1H), 3.51 (m, 1H) , 2.17 (m, 1H), 1.91 (m, 1H) 1.71 (m, 2H); MS (ESH-) ID/Z (relative intensity) 482.2 (M+> +Na, 100), 460.3 (M*- +1, 60),-IR (neat) 3322, 2941, 1727, 1596, 1517,1453, 1428, 1395, 1343 cm'1. [2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl) -4,5-dimethoxy-phenyl]-carbamic acid 9H-fluoren-9-ylmethyl ester (87) The TBDMS ether 81 (t, J= 7.5 Hz, 2H), 6.85 (s, 1H), 4.55-4.38 (m, 3H), 4.3 (m, 1H), 4.05 (m, 1H) , 3.93 (s, 3H), 3.86 (s +m, 4H), 3.7 (m, 1H), 3.59 (m, 1H), 3.45 (m, 1H), 2.2 (m, 1H), 1.92 (m, 1H), 1.82-1.65 (m, 2H); MS (ES+) m/z (relative intensity) 525.2 (M*- +Na, 100), 503.3 (M*- +1, 60); IR (neat) 3328, 2948, 1722, 1596, 1520, 1449, 1394 on'1. ' [2- (2-Hydroxymethyl-pyrrolidine-l-carbonyl)-4,5-dimethoxy phenyl]-carbamic acid allyl ester (88) The TBDMS ether 82 (1.44 g, 3.0 mmol) in THF (30 mL) was deprotected (Method A: Bu^NF (13.6 mL, 3.6 mmol)) to give the product (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless oil (0.996 g, 91%): [oOo24'1- 101.7° (c = 0.35, CHC13) ; *H NMR (CDC13) 6 8.74 (bs, 1H) , 7.79 (s, 1H), 6.82 (a, 1H) , 5,96 (m, 1H), 5.27 (m, 2H), 4.86 (m, 2H), 4.41 (m, 1H), 4.15 (m, 1H), 3.93 (s, 3H), 3.84 (s +m, 4H) , 3.80-3.45 (m, 3H) , 2.2 (m, 1H) , 1.95-1.60 (m, 3H) ; HRMS m/z calcd for Ci8H25N206 365.1713 (M+H) found 365.1699; IR (neat) 3337, 2940, 1728, 1598, 1524, 1455, 1396 cm"1. Cyclisation of monomer alcohols General methods: Method A The alcohol (1 eq), (diacetoxyiodo)benzene (1.1 eq) and TEMPO (0.1 eq) were dissolved in CH2C12 and the mixture stirred at room temperature until reaction was complete (TLC). The reaction mixture was washed with satd NaHS03 ,aq) (xl) and the NaHS03 portion was then washed with CH2C12 (x4) . The combined organic extracts were washed with satd NaHC03 ,aq) (x2), satd NaCl (aq) (xl), dried (MgSO Method B The alcohol (1 eq), pyridinium dichromate (1.2 eq) and 4A molecular sieves (0.5 g/mmol alcohol) in anhydrous CH2Cl2 were stirred at room temperature under a N2 atmosphere until reaction was complete (TLC). The reaction mixture was filtered through celite, washing with EtOAc. The solvent was evaporated in vacuo and the product purified by flash column chromatography. (HS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10fll,lla-hexahydro-5H-pyrrolo[2,l-c][l,4Jbenzodiazepine-10-carboxylic acid benzyl ester (23) The alcohol 16 (0.136 g, 0.33 iranol) was reacted (Method A) with DAIB (0.127 g, 0.39 iranol) and TEMPO (0.005 g, 0.033 mmol) in CH2C12 (6 mL). The product was obtained (flash column chromatography 90% EtOAc/10% n-hexane) as a white foam (0.095 g, 70%) . [ct]D 20-2 + 195.7° (c = 0.23, CHC13), aH NMR (CDC13) 6 7.30 (m, 3H), 7.24 (m, 3H), 6.52 (s, 1H), 5.64 (m, 1H), 5.50 (d, J= 12.3 Hz, 1H), 4.85 (d, J=12.3Hz, 1H) , 3.92 (s, 3H), 3.70 (m, 5H), 3.60 (m, 1H), 3.50 (m, 1H), 2.15 (m, 2H), 2.00 (m, 2H). IR (neat) 3563, 3294, 2953, 1698, 1597, 1573, 1515 cm"1. (llS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2f3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid 4-methoxy-benzyl ester (24) The alcohol 17 (0.214 g, 0.48 mmol) was reacted (Method A) with DAIB (0.175 g, 0.55 mmol) and TEMPO (0.007 g, 0.048 mmol) in CH2Cl2 (6 mL). The product was obtained (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless foam (0.201 g, 94%) . [a]D21-5 + 185.8* (c - 0.22, CHC13) , *H NMR (CDC13) 5 7.27 (s, 1H), 7.17 (d, J- 7.5 Hz, 2H), 6.83 (d, J= 8 Hz, 2H) , 6.49 (s, 1H) 5.62 (m, 1H) 5.32 (d, J - 12 Hz, 1H) , 4.79 ( d, J -12 Hz, 1H) , 3.92 (s, 3H) , 3.78 (s, 3H) , 3.70 (m, 4H) , 3.55 (m, 1H), 3.45 (m, 1H), 2.15 (m, 2H), 2.00 (m, 2H), IR (neat) 3369, 2958, 1705, 1606, 1515 cm'1. (llS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrro2o[l,2-c][If4]benzodiazepine-10-carboxylic acid 2-trimethylsilanyl-ethyl ester (25) O The alcohol 18 (0.110 g, 0.26 mmol) was reacted (Method A) with DAIB (0.092 g, 0.285 mmol) and TEMPO (0.004 g, 0.026 mmol) in CH2C12 (3 mL). The product was obtained (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless oil (0.106 g, 97%). [a]D21-9 + 145.0° (c = 0.21, CHC13) , aH NMR (CDC13) 6 7.23 (S,1H), 6.66 (s, 1H), 5.62 (m, 1H), 4.25 (m, 1H), 4.15 (m, 1H), 3.92 (s, 3H), 3.88 (s, 3H), 3.70 (m, 2H), 3.50 (m, 2H), 2.15 (m, 2H), 2.10 (m, 2H) , 0.9 (m, 2H) , 0.0 (s, 9H) . IR (neat) 3370, 2953, 1704, 1623, 1605, 1515. cm"1. (IIS,llaS) -7,8-Dimethoxy-ll~hydroxy-5-oxo-l,2,3,10fll,lla-hexahydro-5H-pyrrolo[l,2-c][If4]benzodiazepine-10-carboxylic acid 3-(4-nitro-phenyl)-allyl ester (26) (Figure Removed) O The alcohol 19 (0.253 g, 0.52 mmol) was reacted (Method A) with DAIB (0.185 g, 0.57 mmol) and TEMPO (0.008 g, 0.052 mmol) in CH2C12 (4 mL). The product was recrystallased (EtOAc/n-hexane) to give a pale yellow solid (0.24 gf 94%). [ot]D"-8+ 172.3° (c = 0.21, CHC13), *H NMR (CDC13) 6 8 .17 (d, J = 8 . 5 Hz, 2H) , 7 . 44 (d, J= 7.9 Hz, 2H) 7.27 (s, 1H) 6.71 (s, 1H), 6.47 (m 1H), 6.3 (m, 1H), 5.67 (ra, 1H) , 4.77 (s, 2H) , 3.94 (a, 3H) , 3.85 (s, 3H) , 3.70 (m, 2H) , 3.60 (m, 1H), 3.55 (m, 1H) , 2.15 (m, 2H) , 2.05 (m, 2H) , IR (neat) 3369, 2957, 1709, 1602, 1516. cm"1 (HS,llaS) -7, 8-D±methoxy-ll-hydroxy-5-oxo-l, 2 ,3,10,ll hexahydro-5H-pyrrolo[l,2-c] [If4]benzodiazepine-10-carboxylic acid 2-benzenesulfonyl-ethyl ester (27) The alcohol 20 (0.102 g, 0.21 iranol) was reacted (Method A) with DAIB (0.080 g, 0.25 mmol) and TEMPO (0.003 g, 0.021 mmol) in CH2Cl2 (5 mL). The product was obtained (flash column chromatography 80% EtOAc/20% n-hexane) as a colourless oil (0.072 g, 72%). [a]D">3+ 127.5* (c = 0.22, CHC13) , aH NMR (CDC13) 6 7.75 (m, 5H), 7.22 (m, 1H), 6.95 (m, 1H), 5.65 (m, 1H), 4.70 (m, 2H), 3.95 (s, 6H), 3.60 (m, 5H), 3.25 (m, 1H), 2.15 (m, 2H), 2.05 (m, 2H), IR (neat) 3369, 2971, 1710, 1622, 1604, 1516. cm'1 (US, llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l, 2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c][l,4]benzodiazepine-10-carboxylic acid 4,5-dimethoxy-2-nitro-benzyl ester (28(Figure Removed) The alcohol 21 (1.70 g, 3.3 mmol) was reacted (Method B) with pyridinium dichromate (1.47 g, 3.9 mmol) and 4A molecular sieves (1.63 g) in anhydrous CHzCl2 (50 mL). The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a yellow foam (1.04 g, 62%) . [a]26D +99.0° (c = 1.0, CHC13); >H NMR (CDC13) 6 7.65 (s, 1H) , 7.25 (s, 1H), 6.83 (s, 1H) , 6.51 (s, 1H) , 5.74 - 5.72 (m, 1H) , 5.51 (d, J= 15.4 Hz, 1H), 5.44 (d, J= 15.5 Hz, 1H), 4.51 (bs, 1H), 3.92 - 3.87 (m, 9H), 3.68 (s, 3H), 3.54 - 3.50 (m, 2H), 2.16 -2.02 (m, 4H); 13C NMR (CDC13) 6 166.8, 155.4, 153.8, 150.9, 148.5, 148.0, 138.8, 128.2, 126.8, 126.1, 112.5, 110.3, 109.2, 107.9, 86.1, 65.3, 60.1, 56.3, 56.2, 56.2, 56.1, 46.4, 28.6, 23.0; MS (FAB) m/z 650 (M+Cs), 540 (M+Na, 20), 518 (M+l); IR (neat) 3362, 2941, 2616, 1715, 1620, 1523, 1436, 1284, 1134, 1104, 1068, 969, 924, 873, 837, 792, 768, 736, 684, 646 cm'1; HRMS (FAB) m/z calcd for C24H28N3010 518.1761 (M+H) found 518.1775. (llS,llaS)-7,8-dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[1,2-c][1,4]benzodiazepine-10-carboxylic acid 5-methoxy-4- (3-methoxycarbonyl-propoxy)-2-nitro-benzyl ester (29) The alcohol 22 (1.06 g, 1.7 mmol) was reacted (Method B) with pyridinium dichromate (0.76 g, 2.0 mmol) and 4A molecular sieves (0.84 g) in anhydrous CH2C12 (70 mL). The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a yellow foam (0.553 g, 52%). [a]25D +92.5° (c = 1.0, CHC13) ; XH NMR (CDC13) 6 7.64 (s, 1H) , 7.25 (s, 1H), 6.82 (s, IE), 6.50 (s, 1H), 5.74 - 5.71 (m, 1H), 5.49 (d, J - 15.3 Hz, 1H), 5.44 (d, J- 15.0 Hz, 1H), 4.17 - 4.05 (m, 4H), 3,96 - 3.83 (m, 7H), 3.73 - 3.65 (m, 6H), 3.59 - 3.48 (m, 2H), 2.58 - 2.50 (m, 2H) , 2.19 - 2.10 (m, 4H) ; 13C NMR (CDC13) 6 172.3, 170.1, 165.8, 154.4, 153.2, 149.9, 147.5, 146.2, 137.7, 127.3, 125.1, 111.5, 109.4, 108.5, 108.4, 85.2, 67.2, 64.3, 55.2, 55.1, 50.7, 45.4, 29.2, 27.7, 23.2, 22.1; MS (FAB) m/z 626 (M+Na), 604 (M+l); IR (neat) 3385, 2955, 2616, 1734, 1638, 1523, 1285, 1104, 1066, 970, 942, 873, 836, 818, 789, 767, 735, 683, 646 cm"1. 2-{3-[4- ((llS,llaS)-7,8-dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c][l,4]benzodiazepine-10-carbonyloxymethyl)-phenyl]-ureido}-pentanedioic acid diallyl ester (38) The alcohol 37 (0.99 g, 1.45 mmol) was reacted (Method B) with pyridinium dichromate (0.66 g, 1.74 mmol) and 4A molecular sieves (0.87 g) in anhydrous CH2C12 (9 mL). The product was obtained (flash column chromatography 90% EtOAc/10% n-hexane) as a white foam (0.565 g, 57%). [a]D25'6+ 90.0° (c = 0.19, CHC13) , *H NMR (CDC13) 5 7.40 (s, 1H), 7.15 (m, 5H), 6.85 (s, 1H), 5.90 (m, 3H), 5.65 (m, 1H) , 5.35 (m, 5H), 4.85 (d, J = 11.4 Hz, 1H), 4.60 (m, 4H), 4.25 (m, 1H), 4.11 (d, 7.1 Hz, 1H), 3.91 (m, 3H), 3.60 (m, 7H) , 2,5 (m, 2H), 2.25 (m, 1H), 2.05 (m, 4H). HRMS m/z calcd for 681.2772 (M+H) , found 681.2755. 2-[4-( (llS,llaS)-7,8~dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c] [l,4']benzodiazepine-10-carbonyloxymethyl) -phenoxycarbonylamino]-pentanedioic acid diallyl ester (45) The alcohol 44 (0.55 g, 0.8 mmol) was reacted (Method B) with pyridinium dichromate (0.36 g, 0.96 mmol) and 4A molecular sieves (0.4 g) in anhydrous CH2C12 (4 ml). The product was obtained (flash column chromatography 80% EtOAc/20% n-hexane) as a white foam (0.36 g, 66%). 1E NMR (CDC13) 6 7.22 (m, 3H), 7.07 (d, J= 7.5 Hz, 2H), 6.50 (s, 1H), 5.95 (m, 3H), 5.70 (dd, J = 3, 10 Hz, 1H); 5.30 (m, 5H), 4.90 (d, J = 13 Hz, 1H), 4.60 (m, 4H), 4.45 (m, 1H), 3.92 (s, 3H), 3.55 (m, 8H), 2.50 (m, 2H), 2.30 (m, 1H), 2.10 (ra, 4H). (llS,llaS)-7f8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid methyl ester (89) The alcohol 83 (0.41 g, 1.2 mmol) was reacted (Method A) with DAIB (0.47 g, 1.45 mmol) and TEMPO (0.02 g, 0.12 mmol) in CH2C12 (30 mL). The product was obtained (trituration with Et20) as a white solid (0.34 g, 84%): [a]D 2a'3 + 173.6* (c = 0.24, CHC13) ; JH NMR (CDC13) 6 7.3 (s, 1H), 6.68 (s, 1H), 5.66 (m, 1H), 3.96 (s, 3H), 3.91 (s, 3H), 3.85-3.65 (m, 4H), 3.6 (m, 1H), 3.49 (m, 1H), 2.14 (m, 2H), 2.02 (m, 2H), 1.85 (bs, 1H); MS (ES+) m/z (relative intensity) 337.1 (MH' +1, 100); IR (neat) 3216, 2957, 1719, 1604, 1519, 1477, 1437, 1316 cm"1. (HS,llaS)-7,8-Diniethoxy-ll-hydroxy-5-oxo-l,2f3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid tert-butyl ester (90) The alcohol 84 (0.23 g, 0.6 iranol) was reacted (Method A) with DAIB (0.234 g, 0.73 mmol) and TEMPO (0.009 g, 0.06 iranol) in CH2C12 (10 mL). The product was obtained (flash column chromatography EtOAc) as a white foam (0.205 g, 89%): [cx]D20-9+ 162.4° (c = 0.19, CHC13); *H NMR (CDC13) 8 7.22 (s, 1H), 6.62 (s, IE), 5.45 (dd, J= 3.2 Hz, 9.5 Hz, 1H), 3.93 (s, 3H), 3.89 (s, 3H), 3.8-3.69 (m, 2H), 3.58 (in, 1H) , 3.45 (m, 1H) , 2.12 (m, 2H) , 2.0 (m, 2H) , !.39 (s, 9H); (ES+) m/z (relative intensity) 379.1 (M+> +1, 100); IR (neat) 3300, 2970, 1701, 1603, 1513, 1432, 1323 cm'1. (HS,llaS)-7,8-Diniethoxy-ll-hydroxy-5-oxo-l,2,3,10fll,lla-hexahydro-5H-pyrrolo[lr2-c][l,4Jbenzodiazepine-10-carboxylic acid 2,2,2-trichloro-ethyl ester (91) acohol 85 (0.715 g, 1.57 mmol) was reacted (Method A) with DAIB (0.61 g, 1.88 mmol) and TEMPO (0.025 g, 0.16 mmol) in CH2C12 (30 mL). The product was obtained (crystallised from Et20) as a white solid (0.56 g, 78%) : [a]D 20'7 + 129.2' (c = 0.24, CHC13) ; *H NMR (CDC13) 6 7.25 (s, 1H), 6.80 (s, 1H), 5.66 (dd, J - 4.22 Hz, 9.8 Hz, 1H), 5.26 (d, J = 12 Hz, 1H), 4.23 (d, J = 12 Hz, 1H), 3.93 (s, 3H), 3.91 (s, 3H), 3.79 (d, J= 4.4 Hz, 1H), 3.7 (m, 1H), 3.5 (m, 2H), 2.14 (m, 2H), 2.02 (m, 2H); (ES+) m/z (relative intensity) 452.9 (M+- +1, 100); IR (neat) 3307, 2958, 1722, 1617, 1599, 1512, 1453, 1432, 1408 cm'1. (115,1138)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,11,lla-hexahydro-5H-pyrrolo[l,2-c][1,4]benzodiazepine-10-carboxylic acid 4-nitro-benzyl ester (92) (Figure Removed) The alcohol 86 {0.7 g, 1.5 mmol) was reacted (Method A) with DAIB (0.585 g, 1.82 mmol) and TEMPO (0.024 g, 0.15 mraol) in CH2C12 (40 mL) . The product was obtained (flash column chromatography EtOAc) as a yellow foam (0.49 g, 71%): [a]D 21-3 + 174° (c = 0.2, CHC13); aH NMR (CDC13) 6 8.16 (d, J= 8.3 Hz, 2H) , 7.32 (d, J = 8.0 Hz, 2H) , 6.63 (s, 1H), 5.66 (dd, J= 4.3 Hz, 9.6 Hz, 1H), 5.32 (d, J= 13.5 Hz, 1H), 5.08 (d, J= 13.6 Hz, 1H), 3.94 (s, 3H), 3.80 (s, 3H), 3.7 (m, 1H), 3.51 (m, 3H), 2.14 (m, 2H), 1.99 (m, 2H); (ES+) m/z (relative intensity) 480.0 (M+1 +Na, 38) 458.1 (M4" +1, 100); IR (neat) 3312, 2968, 1709, 1602, 1513, 1431, 1402 cm'1. (HS,llaS)-7,8-Dimethoxy-l 1 -hydroxy-5-oxo-l ,2f3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c] [1,4Jbenzodiazepine-10-carboxylic acid 9H-fluoren-9-ylmethyl ester (93) The alcohol 87 (0.735 g, 1.46 mmol) was reacted (Method A) with DAIB (0.565 g, 1.75 nunol) and TEMPO (0.023 g, 0.15 mmol) in CH2C12 (40 mL). The product was obtained (flash column chromatography EtOAc) as a yellow oil (0.64 g, 881): [a]D21'5+ 113.3° (c - 0.20, CHC13) ; :H NMR (CDC13) 6 7.72 (m, 2H) , 7.48-7.0 (m, 7H), 6.68 (s, 1H), 5.7 (m, 1H), 4.55 (m, 1H) , 4.15-3.9 (m, 5H), 3.85-3.65 (m, 4H), 3.65-3.45 (m, 3H), 2.14 (m, 2H), 2.03 (m, 2H); (ES+) m/z (relative intensity) 501.1 (M+' +1, 100); IR (neat) 3307 2961, 1702, 1602, 1512, 1450, 1406 cm"1. (llS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid allyl ester (94) Dimethyl sulfoxide (1.16 g, 1.1 mL, 14.9 mmol, 5 eq) was added to a solution of cyanuric chloride (0.55 g, 3.0 mmol 1.2 eq) in anhydrous THF (20 mL) at -30'c under a nitrogen atmosphere. The solution was stirred at -30°C for 30 min. A solution of the alcohol 88 in anhydrous THF (10 mL) was added dropwise followed by triethylamine ( 1.25 g, 1.73 mL, 12.4 mmol, 5 eq). After 15 min at -30°C the solution was allowed to reach room temperature. The solvent was evaporated in vacuo and the residue dissolved in dichloromethane (50 mL) , extracted with 1M HC1 (3 x 30 mL), H20 (2 x 30 mL), brine (50 mL) , dried (MgSO Deprotection of groups in compounds 38 and 45 2-{3-[4- ((HSfllaS) -7f8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c][l,4]benzodiazepine-10-carbonyloxymethyl)-phenyl]-ureido}-pentanedioic acid (39) A solution of the diallyl ester 38 (0.5 g, 0.74 mmol, 1 eq), Pd(PPh3)4 (0.043 g, 0.037 mmol, 0.05 eq) and pyrrolidine (0.26 g, 0.31 mL, 3.7 mmol, 5 eq) in anhydrous CH2C12 (15 mL) was stirred at room temperature for 1 h and the solvent removed in vacuo. The residue was washed with EtOAc (3 x 15 mL), dissolved in MeOH (5 mL) and passed down an IRC 50, weakly acidic, ion exchange column, eluting with MeOH (100 mL). The solvent was removed in vacuo to give the product as a white foam (0.39 g, 87%). [a]D26'4 + 146.0' (c = 0.21, EtOH) , aH NMR (de DMSO) 5 8.82 (s, 1H) , 7.32 (d, J= 7.12 Hz, 2H) , 7.11 (d, J= 7.52 Hz, 2H) , 7.04 (s, !H) , 6.69 (s, 1H), 6.50 (d, J- 6.6 Hz, 1H), 5.47 (d, J= 9.16 Hz, 1H), 5.14 (d, J = 12 Hz, 1H), 4.75 (d, J = 12 Hz, 1H), 4.11 (m, 1H), 3.78 (m, 5H), 3.69 (s, 3H), 3.40 (m, 4H), 2.27 (t, J= 7.5 Hz, 2H) , 1.95 (m, 4H) . HRMS m/z calcd for C28H33N40U 601.2146 (M+H), found 601.2160. 2-[4-((llS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10/llrlla-hexahydro-5H-pyrrolo[2fl-c][1,4]benzodiazepine-10-carbonyloxymethyl)-phenoxycarbonylamino]-pentanedioic acid (46) A solution of the diallyl ester 45 (0.596 g, 0.95 iranol, 1 eq), Pd(PPh3)4 (0.055 g, 0.047 mmol, 0.05 eq) and pyrrolidine (0.34 g, 0.39 mL, 4.7 mmol, 5 eq) in anhydrous CHjClz (18 mL) was stirred at room temperature for 1 h and the solvent removed in vacua. The residue was washed with EtOAc (3 x 15 mL), dissolved in MeOH (15 mL) and passed down an IRC 50, weakly acidic, ion exchange column, eluting with MeOH (80 mL). The solvent was removed in vacua to give the product as a white foam (0.46 g, 88%). aH NMR (df DMSO) 6 7.95 (d, J = 6.25 Hz, 1H), 7.33 (d, J= 8 Hz, 2H), 7.10 (m, 3H) 6.73 (s, 1H), 5. 49 (d, J= 9.2 Hz, 1H), 5.2 (d, J = 12.5 Hz, 1H), 4.89 (d, J= 12.75 Hz, 1H), 4.00 (m, 1H), 3.80 (m, 7H), 3.45 (m, 4H) 2.30 (t, J = 7.5 Hz, 2H), 1.95 (m, 5H). MS (ES+) 602.2. Deprotection of N-10 protected monomers to give (llaS)-7,8-dimethoxy-1,2,3,lla-tetrahydro-5H-pyrrolo12,1-c][1,4]ben2odiazepine-5-one (30) O Deprotection of (115,llaS)-7,8~Dimethoxy-ll-hydroxy-5-oxo- l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid 4-methoxy-benzyl ester (24) The N-10 protected PBD (0.08 g, 0.18 mmol) was dissolved in 10% TFA/CH2C12 (4 ml) at 0°C. The solution was stirred at 0°C for 25 min then poured onto ice and neutralised with satd NaHC03 {Bq). The aqueous portion was extracted with CH2Cl2 (3 x 20 mL) and the combined organic extracts were washed with H20 (50 mL), satd NaCl (aq) (50 mL), dried (MgS04) and evaporated in vacuo. Purification by flash column chromatography (3% MeOH/97% CHC13) gave the product 30 as a yellow solid (0.022 g, 47%). 1H NMR (CDC13) 6 7.61 (d, J = 4.3 Hz, 1H), 7.46 (s, 1H), 6.72 (s, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.75 (m 2H) , 3.65 (m, 1H), 3.5 (m, 1H), 2.30 (m 2H), 1.9 (m, 2H). Deprotection of (HS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-1,2,3,10,11,lla-hexahydro-5H-pyrrolo[l,2-c][1, 4]benzodiazepine-10-carboxylic acid 2-trimethylsilanyl-ethyl ester (25) A 1.0 M THF solution of tetra-N-butyl-ammonium fluoride (0.295 mL, 0.295 mmol, 1.2 eq.) was added via syringe to a solution of the N-10 protected PBD (0.104 g, 0.25 mmol, 1 eq) in THF (5 mL) at 0°C. The reaction mixture was stirred at room temperature for 4 h. The solvent was removed in vacuo and the product purified by flash column chromatography (4% MeOH/CHCl3) to give the product 30 as a yellow solid (0.052 g, 81%). Deprotection of (IIS,llaS)-7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid 3- (4-nitro-phenyl)-allyl ester (26) A solution of the N-10 protected PBD (0.12 g, 0.25 mmol, 1 eq), Pd(PPh3) The N-10 protected PBD (0.05 g, 0.1 mmol, 1 eq) was dissolved in MeOH (5 mL) and 10% Palladium on carbon (0.015 g 30 wt%) and cyclohexadiene (0.016 g, 0.018 mL, 0.2 mmol, 2 eq) were added. The mixture was heated at reflux for 5 h, cooled, filtered through celite and the solvent evaporated in vacuo. Purification by flash column chromatography (3% MeOH/97% CHC13) gave the product 30 as a yellow solid (0.017 g, 65%). Deprotection of (HS,llaS) -7,8-Dimethoxy-ll-hydroxy-5-oxo-l,2f3f10,ll,lla-hexahydro-5H-pyrrolo[l,2-c][l,4]benzodiazepine-10-carboxylic acid 2-benzenesulfonyl-ethyl ester (27) A 2.0 M MeOH solution of dimethylamine (0.18 mL, 0.37 mmol, 3 eq.) was added via syringe to a solution of the N-10 protected PBD (0.06 g, 0.122 mmol, 1 eq) in MeOH (I mL). The reaction ixture was stirred at room temperature for 18 h. The solvent was removed in vacua and the product purified by flash column chromatography (5% MeOH/CHCl3) to give the product 30 as a colourless oil (0.028 g, 87%). Deprotection of (HS,llaS) -7,8-Dimethoxy-ll-hydroxy-5-oxo-1,2,3,10,11,1la-hexahydro-5H -pyrrole[2,1-c][1,4]benzodiazepine-10-carboxylic acid 4,5~dimethoxy-2-nitro-benzyl ester (28) A solution of the N-10 protected PBD (0.25 g) in MeOH (10 mL ) was irradiated at 365 nM. The reaction was monitored by reversed phase HPLC (CIS column, 5 uM particle size, 250 mm x 4.6 mm; mobile phase 70% H20/30% CH3CN/0.1% TFA; detection at 245 nM) against a standard sample of the parent PBD 30. Aliquots of the reaction mixture (20 pL) were injected at 1 h intervals. The conversion to parent PBD was complete in 12 h. MS (ES+) m/z 261 (M+-) Deprotection of 2-[4-((HS,llaS) -7,8-Dimethoxy-ll-hydroxy-5-oxo-1,2,3,10,11,1 la-hexahydro-5H-pyrrolo[2,l-c] [l,4]benzodiazepine-10-carbonyloxymethyl) -phenoxycarbonylamino]-pentanedioic acid (46) An aliquot of a 10 mM stock solution of the carbamate monomer prodrug was diluted to 100 uM in Carboxypeptidase G2 (CPG2) assay buffer (100 mM Tris-HCl, pH 7.3; 260 uM ZnCl2) . CPG2 (1 unit) was added and the reaction was incubated at 37°C. The reaction was monitored by reversed phase HPLC (C18 column, 5 uM particle size, 250 mm x 4.6 mm; mobile phase 70% H20/30% CH3CN/0.1% TFA; detection at 245 nM) with 20 uL aliquots being injected at 0, 10, 20, 30, 40, 50 and 60 min. The conversion of the prodrug to the parent imine 30 was complete in 60 minutes. Deprotection of (HS,llaS) -7,8-Dimethoxy-ll-hydroxy-5-oxo-1,2,3,10,11,lla-hexahydro-5H-pyrrolo[1,2-c][1,4]benzodiazepine-10-carboxylic acid allyl ester (94) A solution of the N-10 protected PBD (0.35 g, 0.96 mmol, 1 eq), Pd(PPh3)4 (0.055 g, 0.05 mmol, 0.05 eq) and pyrrolidine (0.025 g, 0.1 mL, 1.2 mmol, 1.25 eq) in anhydrous CH2C12 (12 mL) was stirred at room temperature for 3.5 h and the solvent removed in vacua. The product was purified by flash column chromatography (4% MeOH/96% CHC13) to give 30 as a yellow oil (0.176 g, 71%). (Figure Removed)Oxalyl chloride (12.44 g, 8.55 mL, 97.9 mmol, 2.5 eq) and DMF (cat) were added to a solution of dimer core 53 (19.37 g, 39.2 mmol, 1 eq) in anhydrous THF (200 ml) under a N2 atmosphere. The solution was stirred at room temperature for 18 h. The resultant solution was added dropwise to a solution of (S)-(+)-2-pyrrolidine-methanol (9.89 g, 9.65 mL, 97.9 mmol, 2.5 eq) and triethylamine (16.62 g, 22.89 mL, 164 mmol, 4.2 eq) in anhydrous THF (150 mL) at -25°C (dry ice/ethylene glycol) under a N2 atmosphere. The reaction mixture was allowed to come to room temperature and stirred for 18 h. The solvent was removed in vacua and the residue dissolved in CH2C12 (750 mL), washed with 1 M HC1 (3 x 200 mL) , satd NaHC03 ,aq, (3 x 200 mL) , H20 (2 x 200 mL) , satd NaCl(aq) (250 mL) , dried (MgS04) and evaporated in vacuo. The product was purified by flash column chromatography (5% MeOH/95% EtOAc-10% MeOH/90% EtOAc) to give a yellow foam (11.81 g, 45.6%). [a]D24'7 -94.1° (c = 0.260, CHC13) ; XH NMR (CDC13) 6 7.69 (s, 2H) , 6.81 (s, 2H), 4.5-4.35 (m, 4H), 4.15 (t, J = 6.6 Hz, 4H), 3.98 (s, 6H), 3.9 (m, 2H), 3.8 (m, 2H), 3.18 (t, J= 6.9 Hz, 4H), 2.2 (m, 2H), 2.0 (m, 4H), 1.9-1.65 (m, 8H); IR (neat) 3400, 2946, 2873, 1618, 1577, 1522, 1276 cm"1; HRMS m/z calcd for C3iH4iN4012 661.2721 (M+H) found 661.2690. (2S)-!,!'-[[(Pentane-l,5-diyl)dioxy]bis[(2-nitro-5-methoxy-l,4-phenylene)carbonyl]Jbis[2~(tert- A solution of t-butyldimethylsilyl chloride (6.59 g, 43.56 mmol, 2.6 eq), imidazole (5.7 g, 83.8 mmol, 5 eq) and dimer nitro-alcohol 54 (11.07 g, 16.75 mmol, 1 eq) in anhydrous DMF (30 mL) was stirred at room temperature under a N2 atmosphere for 96 h. The reaction mixture was diluted with H20 (500 mL) and extracted with Etoac (4 x 250 mL). The combined organic extracts were washed with H2O (2 x 250 mL) , satd NaCl(aq, (250 mL) , dried (MgSO4) and evaporated in vacuo. Purification by flash column chromatography (60% EtOAc/40% n-hexane) gave the product as an off white foam (9.02 g, 60.5%). [a]D24'9 -84.9° (c = 0.212, CHC13); 1E NMR (CDC13) 6 7.68 (s, 2H) , 6.76 (s, 2H), 4.37 (m, 2H), 4.13 (m, 4H), 3.94 (s, 6H), 4.0-3.85 (m, 4H), 3.12 (m, 4H), 2.15-1.9 (m, 4H), 0.91 (s, 18H), 0.12 (s, 12H); IR (neat) 2952, 2857, 1737, 1643, 1577, 1522 cm"1; HRMS m/z calcd for 889.4451 (M+H) found 889.4473. (2S) -!,!'-[[ (Pentane-1 ,5-diyl) dioxy]bis [ (2-amino-5-methoxy-l ,4-phenylene)carbonyl]]bis[2-(tert-bvtyldimethylsilyloxymethyDpyrrolidine] (56) A solution of the dimer nitro compound 55 (9.0 g, 10.12 mmol) in ethanol (100 mL) was hydrogenated (Parr apparatus) over 10% Palladium on carbon (0.9 g, 10 wt%), maintaining the H2 pressure at 16 psi. The reaction was complete when no more H2 was consumed. The mixture was filtered through celite and the ethanol evaporated in vacuo. Purification by flash column chromatography (90% EtOAc/10% n-hexane) gave the product as a yellow foam (6.1 g, 73%). [a]D25-5 -141.2° (c = 0.241, CHC13) ; XH NMR (CDC13) 6 6.76 (s, 2H), 6.23 (s, 2H), 4.5-4.3 (m, 4H), 3.99 (t, J= 6.6 Hz, 4H) 3.75 (s, 6H) , 3.67 (m, 2H),.3.53 (m, 4H), 2.05 (m, 4H), 1.95 (m, 6H), 1.7 (m, 8H), 0.9 (s, 18H), 0.04 (s, 12H); IR (neat) 3449, 3349, 2952, 2857, 1624, 1592, 1514, 1406 cm'1; HRMS m/z calcd for 829.4967 (M+H) found 829.4998. (2S) -!,!'-[[ (Pentane-1,5-diyl) dioxy]bis[ (2-isocyanato-5-methoxy-1,4-phenylene) carbonyl] ]bis [2- (tert-butyldimethylsilyloxymethyl)pyrrolid±ne] (57) A solution of triethylamine (2.7 eq.) in anhydrous toluene was added to the amine (56)(1 eq.) and triphosgene (0.72 eq.) in anhydrous toluene under a N2 atmosphere. The reaction was finished after 2 hours, (monitored by IR, VNCO 2265 cm"1). The product was used without further purification. Synthesis of Dimer Carbamates General Method(Figure Removed)A solution of the appropriate alcohol (2 eq) and triethylamine (2.2 eq) in either anhydrous toluene or anhydrous CH2C12 was added dropwise to a solution of the isocyanate (57)(1 eq) in anhydrous toluene. The reaction was monitored by IR (disappearance of the VNCO 2265 cm"1 peak) . The reaction mixture was filtered and the filtrate evaporated in vacuo. The product was purified by flash column chromatography. (25) -!,!'-[[ (Pentane-1,5-diyl)dioxyjbis[ (2-allyloxycarbonylamino-5-methoxy-lf4-phenylene) carbonyl]]bis[2- (tert-butyldimethylsilyloxymethyljpyrzolidine] (58) (Figure Removed) A solution of anhydrous allyl alcohol (0.09 g, 0.105 mL, 1.54 mmol, 2.4 eq) and triethylamine (0.143 g, 0.2 mL, 1.41 nunol) in anhydrous toluene (4 mL) was added to a solution of the isocyanate 57 prepared from the dimer amine 56 (0.533 g, 0.64 mmol), triphosgene (0.14 g, 0.46 mmol) and triethylamine (0.175 g, 0.24 mL, 1.74 mmol) in anhydrous toluene (20 mL). The reaction was complete in 16 h. The product was obtained (flash column chromatography 60% EtOAc/40% n-hexane) as a yellow oil (0.4 g, 62%) . [alt,26'1 -105.7° (c = 0.227, CHC13) ; aH NMR (CDC13) 6 9.18 (s, 2H) , 7.85 (s, 2H), 6.83 (s, 2H), 6.0 (m, 2H), 5.3 (m, 4H), 4.65 (m, 4H), 4.35 (m, 2H) , 4.15 (m, 4H), 4.05 (m, 2H), 3.8 (s, 6H) , 3.7 (m, 2H), 3.5 (m, 4H) , 2.05 (m, 4H), 1.95 (m, 6H), 1.7 (m, 4H), 0.9 (s, 18H), 0.07 (s, 12H); IR (neat) 3306, 2952, 2930, 2857, 1731, 1621, 1598, 1523, 1406 cm"1; HRMS m/z calcd for C5iH8iN4Oi2Si2 997.5390 (M+H) found 997.5336. (2S) -1,1'-II(Pentane-1,5-diyl)dioxy]bis[(2-[N- (4- (diprop-2-enyl-L-glutamylcarbonyloxy) benzyloxycarbonyl)]amino-5-methoxy-l,4-phenylene)carbonyl]]bis[2-Ctert-butyldimethylsilyloxymethyDpyrrolidine] (62) -i A solution of the carbamate progroup 42 (1.27 g, 3.4 mmol) and triethylamine (0.37 g, 0.515 mL,) in anhydrous CH2C12 (25 mL) was added to a solution of the isocyanate 57 prepared from the diraer amine 56 (1.4 g, 1.7 iranol), triphosgene (0.36 g, 0.12 iranol) and triethylamine (0.46 g, 0.64 mL, 4.55 mmol) in anhydrous toluene (75 mL). The reaction was complete in 48 hours. The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a white foam (1.65 g, 60%). [a]D22t7 -39.13° (c = 0.23, CHC13); aH NMR (CDC13) 6 9.24 (s, 2H) , 7.84 (s, 2H), 7.4 (m, 4H), 7.1 (m, 4H), 6.82 (s, 2H), 5.95 (m, 4H), 5.8 (d, J- 7.6 Hz, 2H), 5.3 (m, 8H), 5.16 (d, J - 12.4 Hz, 2H), 5.1 (d, J = 12.3 Hz, 2H), 4.65 (m, 8H), 4.5 (m, 2H), 4.35 (m, 2H), 4.15 (m, 4H), 4.05 (m, 2H), 3.8 (s, 6H), 3.7 (m, 2H), 3.5 (m, 4H), 2.55 (m, 4H), 2.3 (m, 2H), 2.05 (m, 6H), 1.95 (m, 4H), 1.6 (m, 6H), 0.9 (s, 18H), 0.07 (s, 12H); IR (neat) 3338, 2952, 2857, 1738, 1648, 1617, 1597, 1523 cm (2S) -1,1'-[ [ (Pentane-1,5-diyl) dioxyjbis[ (2-[N- (4- (diprop-2-enyl-L-glutamylcarbonylamino)benzyloxycarbonyl) ]amino~5-methoxy-l, 4-phenylene)carbonyl]]bis[2-(tert-butyldimethylsilyloxymethyl)pyrrolidine] (63) A solution of the urea progroup 35 (1.27 g, 3.4 nunol) and triethylamine (0.37 g, 0.515 mL,) in anhydrous CH2C12 (25 mL) was added to a solution of the isocyanate 57 prepared from the dimer amine 56 (1.4 g, 1.7 iranol), triphosgene (0.36 g, 0.12 mmol) and triethylamine (0.46 g, 0.64 mL, 4.55 mmol) in anhydrous toluene (75 mL). The reaction was complete in 48 h. The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a white foam (1.65 g, 60%). [cOo23'1 -50.7° (c = 0.217, CHC13); 2H NMR (CDC13) 6 9.0 (s, 2H) , 7.68 (s, 2H) , 7.25 (m, 10H), 6.8 (s, 2H), 5.9 (m, 6H), 5.25 (m, 8H), 5.05 (m, 4H), 4.6 (m, 10H), 4.35 (m, 2H), 4.15 (m, 2H), 4.05 (m, 4H), 3.78 (s, 6H) , 3.65 (m, 2H) , 3.5 (m, 4H), 2.45 (m, 4H), 2.2 (m, 2H), 2.1-1.8 (m, 10H), 1.7 (m, 6H), 0.9 (s, 18H), 0.07 (s, 12H); IR (neat) 3351, 2952, 1736, 1700, 1666, 1601, 1521, 1411, 1200 cm'1. (25) -!,!'-[[ (Pentane-1,5-diyl)dioxy]bis[ (2- (2~ phenylthioethyloxycarbonyl) -amino~5-methoxy~l,4-phenylene)carbonyl]bis[2-(tert-butyldimethylsilyloxymethyl) pyrrolidine] (95) A solution of 2-(phenylthio) ethanol (0.65 mL, 4.8 ininol,) and triethylamine (0.74 mL, 5.3 ininol) in anhydrous toluene (10 mL) was added to a solution of the isocyanate 57 prepared from the dimer amine 56 (2.0 g, 2.4 mmol), triphosgene (0.515 g, 1.7 mmol) and triethylamine (0.907 mL, 6.5 mmol) in anhydrous toluene (100 mL). The reaction was complete in 90 h. The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a white foam (0.862 g, 30%): [a]D26 = -2.6* (c = 0.4, CHC13) ; aH NMR (CDC13) 6 9.13 (s, 2H), 7.77 (s, 2H), 7.40-7.32 (m, 4H) , 7.30-7.21 (m, 4H), 7.19-7.10 (m, 2H), 6.79 (s, 2H), 4.32-4.17 (m, 6H), 4.15-3.84 (m, 4H), 3.83-3.32 (m, 14H), 3.23-2.99 (m, 4H), 2.13-1.80 (m, 10H) , 1.78-1.51 (m, 4H) , 0.86 (s, 18H), 0.01 (s, 12H); MS (ES) m/z (relative intensity) 1211 (M+- +Na, 6), 1189 (M+- +1, 100); IR (neat) 2928, 1729, 1597, 1522, 1469, 1406, 1258, 1201 cm" (23) -!,!'-[[(Pentane-1,5-diyl)dioxy]bis[(2-(2-phenylsulfonylethyloxycarbonyl) -amino-5-methoxy-l ,4-phenylene)carbonyljbis[2-(tert-butyldimethylsilyloxymethyDpyrrolidine] (96) A solution of 2-(phenylsulfonyl)ethanol (0.58 mL, 4.8 mraol,) and triethylamine (0.74 mL, 5.3 mraol) in anhydrous toluene (20 mL) was added to a solution of the isocyanate 57 prepared from the dimer amine 56 (2.0 g, 2.4 mmol), triphosgene (0.515 g, 1.7 mmol) and triethylamine (0.907 mL, 6.5 mmol) in anhydrous toluene (100 mL). The reaction was complete in 90 h. The product was obtained (flash column chromatography 50% EtOAc/50% n-hexane) as a white foam (2.17 g, 77%): [a]D22 = -87.5° (c = 1.0, CHC13) ; aH NMR (CDC13) 6 9.04 (s, 2H), 8.07-7.93 (m, 3H), 7.77-7.48 (m, 7H), 7.26 (s, 2H), 6.82 (s, 2H), 4.46-4.26 (m, 5H) , 4.18-3.89 (m, 6H), 3.88-3.64 (m, 8H), 3.63-3.42 (m, 8H), 2.16-1.84 (m, 10H), 1.83-1.51 (m, 5H), 0.90 (s, 18H), 0.04 (s, 12H); MS (ES) m/z (relative intensity) 1256 (M+I 41, 100); IR (neat) 3299, 2952, 1735, 1602, 1528, 1326, 1025, 841 cm'1. Deproteotion of alcohols General methods for the deprotection of dimer tert- butyldimethylsilyl ethers Method A A 1.0 M THF solution of tetra-N-butyl-ammonium fluoride (2.4 eq.) was added via syringe to a solution of the TBDMS ether (I eq.) in THF at 0°C. The reaction was stirred at room temperature until reaction was complete (TLC). The solvent was removed in vacuo and the product purified by flash column chromatography. Method B A solution of the TBDMS ether in a mixture of AcOH/THF/%0 (3/1/1) was stirred at room temperature until reaction was complete (TLC). The reaction mixture was cooled (ice bath) and carefully neutralised with NaHCOs (aq) (1 eq) . The mixture was extracted with EtOAc (x3) the combined extracts were washed with water (xl), satd NaCl (aq) (xl) , dried (MgS04) and evaporated in vacua. The product was purified by flash column chromatography. (25)-!,!'-[[(Pentane-lf5-diyl)dioxy]bis[(2-allyloxycarbonylamino-5-methoxy-l,4-phenylene)carbonyl]]bis[2-(hydroxymethyl)pyrrolidine] (59) The TBDMS ether 58 (0.32 g, 0.32 mmol) in THF (20 mL) was deprotected (Method A: Bu glutamylcarbonyloxy)benzyloxycarbonyl) ]amino-5-methoxy-l,4-phenylene) carbonyl]]bis [ (2-hydroxymethyl)pyrrolidinej (64) 5 The TBDMS ether 62 (1.63 g, 0.99 mmol) was deprotected (Method B: AcOH (18 mL)/THF (6 mL)/H20 (6 mL) then NaHC03 (26.31 g, 313.2 mmol) in H20 (300 mL)) to give the product (flash column chromatography 2% MeOH/98% EtOAc) as a colourless oil (1.0 g, 72%). [a]D22>7 -34.01° (c = 0.147, CHC13) ; aH NMR (CDC13) 6 8.8 (s, 10 2H), 7.7 (s, 2H), 7.4 (m, 4H) , 7.1 (m, 4H) , 6.78 (s, 2H), 5.95 (m, 4H), 5.8 (m, 2H), 5.3 (m, 8H), 5.17 (d, J= 12.4 Hz, 2H), 5.10 (d, J= 12.3 Hz, 2H), 4.65 (m, 8H) , 4.5 (m, 2H), 4.36 (m, 4H), 4.12 (m, 4H), 3.82 (m, 2H), 3.8 (s, 6H), 3.7 (m, 2H), 3.55 (m, 2H), 3.43 (m, 2H), 2.51 (m, 4H), 2.3 (m, 2H), 2.1 (m, 4H) , 15 1.95 (m, 4H), 1.85 (m, 2H), 1.68 (m, 6H); IR (neat) 3326, 2946, 1731, 1597, 1524, 1203 cm"1. (25) -!,!'-[[ (Pentane-1,5-diyl)dioxyjbis[ (2-[N- (4- (diprop-2-enyl-L-glutamylcarbonylamino)benzyloxycarbonyl) ]amino-5-methoxy-l, 4- 20 phenylene) carbonyl]]bis [ (2-hydroxymethyl)pyrrolidine] (65) n H H The TBDMS ether 63 (1.075 g, 0.66 mmol) was deprotected (Method B: AcOH (12 mL)/THF (4 mL)/H20 (4 mL) then NaHC03 (17.54 g, 209.0 mmol) in H20 (250 mL)) to give the product (flash column chromatography 3% MeOH/97% EtOAc) as a white foam (0.69 g, 75%). [a]D21'6 -46.98° (c = 0.3299, CHC13) ; *H NMR (CDC13) 5 8.6 (s, 2H) , 7.72 (m, 4H), 7.21 (m, 8H), 6.79 (s, 2H), 6.02 (d, J= 7.8 Hz, 2H), 5.86 (m, 4H), 5.3 (m, 8H), 5.04 (s, 4H), 4.6 (m, 8H) , 4.49 (m, 2H), 4.39 (m, 2H), 4.01 (m, 4H), 3.83 (m, 2H), 3.78 (s, 6H), 3.69 (in, 2H) , 3.5 (m, 4H) , 2.45 (m, 4H) , 2.2 (m, 2H) , 2.1 (m, 2H), 2.0-1.8 (m, 10 H), 1.7 (m, 4H) , 1.55 (m, 2H); IR (neat) 3351, 2946, 2877, 1732, 1601, 1520, 1202 cm"1. (25) -!,!'-[[ (Pentane-1,5-diyl)dioxyjbis[ (2- (2-phenylthioethyloxycarbonyl)-amino-5-methoxy-l, 4-phenylene) carbonyl]bis [2-hydroxymethylpyrrolidine] (97) (Figure Removed) (25) -!,!'-[[ (Pentane-1,5-d±yl) dioxyjbis [ (2- (2-phenylsulfonylethyloxycarbonyl) -amino-5-methoxy-l, 4-phenylene) carbonyl ]bis [2-hydroxymethylpyrrolidine] (98) The TBDMS ether 95 (0.862 g, 0.7 mmol) was deprotected (Method B: AcOH (6 mL) /THF (2 mL)/H2O (2 mL) then NaHC03 (8.8 g, 104.8 mmol) in H20 (400 mL)) to give the product (flash column chromatography EtOAc) as a pale yellow foam (0.486 g, 70%): [a]D26 -64.0° (c = 1.0, CHC13); XH NMR (CDC13) 6 8.73 (s, 2H) , 7.70 (s, 2H) , 7.46-7.36 (m, 4H), 7.35-7.14 (m, 6H), 6.80 Cyclisation of dimer alcohols General methods Method A The alcohol (1 eq), (diacetoxyiodo)benzene (2.3 eq) and TEMPO (0.23 eq) were dissolved in CH2C12 and the mixture stirred at room temperature until reaction was complete (TLC). The reaction mixture was washed with satd NaHS03 (aq) (xl) and the NaHS03 portion was then washed with CH2Cl2 (x4) . The combined organic extracts were washed with satd NaHCO3 (aq) (x2), satd NaCl The alcohol (1 eq), pyridinium dichromate (2.4 eq) and 4A molecular sieves (0.1 g/mmol alcohol) in anhydrous CH2C12 were stirred at room temperature under a N2 atmosphere until reaction was complete (TLC). The reaction mixture was filtered through celite, washing with EtOAc. The solvent was evaporated in vacua and the product purified by flash column chromatography. l,l'-[[Pentane-l,5-diyl]dioxy]bis[(HS,llaS)-10-(allyloxycaxbonyl)-ll-hydroxy-7-methoxy-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c][1,4]benzodiazepine-5-one] (60) The alcohol 59 (0.227 g, 0.295 nunol) was reacted (Method B) with pyridinium dichromate (0.27 g, 0.71 mmol) and 4A molecular sieves (0.295 g) in anhydrous CH2C12 (5 mL). The product was obtained (flash column chromatography 5% MeOH/95% EtOAc) as a white foam (0.073 g, 32%) . *H NMR (CDC13) 6 7.24 (s, 2H), 6.65 (s, 2H), 5.8 (m, 2H), 5.63 (d, J= 8.7 Hz, 2H), 5.12 (m, 4H) , 4.66 (m, 2H) , 4.44 (m, 2H) , 4.17 (m, 2H), 4.01 (m, 4H), 3.9 (m, 8H), 3.69 (m, 2H), 3.50 (m, 4H), 2.12 (m, 4H), 2.02 (m, 4H), 1.91 (m, 4H), 1.64 (m, 2H); HRMS m/z calcd for C39H48N4Oi2Na 787.3166 (M+Na) found 787.3173. l,l'-[[Pentane-l,5-diyl]dioxy]bis[(US,llaS)-10~[N-(4-(diprop-2-enyl-L-glutamy2carbonyloxy)benzyloxycarbonyl) ]-ll-hydroxy-7-rnethoxy-lf2r3f10fllflla-hexahydro-5H-pyrrolo[2,l-c] [l,4]benzodiazepine-5-one] (66) rhe alcohol 64 (1.0 g, 0.71 mmol) was reacted (Method A) with )AIB (0.53 g, 1.64 mmol) and TEMPO (0.025 g, 0.164 mmol) in CH2C12 [10 mL). The product was obtained (flash column chromatography 5% MeOH/951 EtOAc) as a white foam (0.72 g, 72%). [a]D22'6 +128.03° (c = 0.289, CHC13); JH NMR (CDC13) 5 7.23 (s, 2H) , 7.16 (d, J = 6.9 Hz, 4H), 7.01 (d, J = 6.8 Hz, 4H) , 6.55 (s, 2H), 6.01 (s, 2H), 5.89(m, 4H), 5.63 (m, 2H), 5.3 (m, 10H), 4.83 (d, J- 11.9 Hz, 2H), 4.65 (m, 8H), 4.44 (m, 2H), 4.09 (m, 2H), 3.88 (m, 8H), 3.75 (m, 4H), 3.49 (m, 4H), 2.48 (m, 4H), 2.27 (m, 2H), 2.15-1.95 (m, 10H), 1.89 (m, 4H), 1.52 (m, 2H). [a]D22>6 +128.03° (c = 0.289, CHC13) ; ^ NMR (CDC13) 6 7.23 (s, 2H) , 7.16 (d, J= 6.9 Hz, 4H), 7.01 (d, J= 6.8 Hz, 4H), 6.55 {s, 2H), 6.01 (s, 2H), 5.89(ra, 4H), 5.63 (m, 2H), 5.3 (m, 10H), 4.83 (d, J = 11.9 Hz, 2H), 4.65 (m, 8H), 4.44 (m, 2H), 4.09 (m, 2H), 3.88 (m, 8H), 3.75 (m, 4H), 3.49 (m, 4H), 2.48 (m, 4H), 2.27 (m, 2H), 2.15-1.95 (m, 10H), 1.89 (m, 4H), 1.52 (m, 2H). l,l'-[lPentane-l,5-diyl]dioxy]bis[(llS,llaS)-10-[N-(4-(dipzop-2-enyl-L-glutamylcarbonylamino) benzyloxycarbonyl) ] -11-hydroxy-7-methoxy-l,2f3,10fllflla-hexahydro-5H-pyrrolo[2,l-c] [l,4]benzodiazepine-5-one] (67) The alcohol 65 (0.51 g, 0.36 mmol) was reacted (Method A) with DAIB (0.27 g, 0.83 mmol) and TEMPO (0.012 g, 0.083 mmol) in CH2C12 (8 mL). The product was obtained (flash column chromatography 5% MeOH/95% EtOAc) as a white foam (0.325 g, 64%). [a]D21>1 +192.0° (c = 0.216, CHC13); aH NMR (CDC13) 6 8.03 (s, 2H) , 7.33 (m, 4H) , 7.26 (s, 2H), 7.15 (d, J = 7.6 Hz, 4H), 6.35 (s, 2H), 5.85 (m, 8H), 5.64 (dd, J = 4 Hz, 9.9 Hz, 2H), 5.25 (m, 10H), 4.6 (m, 12H), 4.02 (m, 2H), 3.87 (m, 8H) , 3.8 (m, 2H) , 3.72 (m, 2H) , 3.57 (m, 4H), 2.69 (m, 2H), 2.36 (m, 4H), 2.2-1.95 (m, 10H), 1.68 (m, 2H), 1.5-1.2 (m, 4H); IR (neat) 3359, 2949, 1737, 1707, 1603, 1547, 1515 cm'1. !,!'-[[Pentane-1,5-diyl]dioxy]bis[(llS,llaS)-10-[N-(2-phenylthioethyloxycarbonyl) ] -ll-hydroxy-7-methoxy-1,2,3,10,11,lla-hexahydro~5H-pyrrolo[2,1-c][1,4]benzodiazepine-5-one] (99) (Figure Removed)-i Dess-Martin periodinane (0.15% v/v soln. in DCM, 1.55 mL, 0.55 iranol) was added to a solution of the dimer alcohol 97 (0.2.43 g, 0.25 iranol) in anhydrous DCM (10 mL). The reaction was monitored by HPLC and was complete in 1.5 h. The reaction mixture was washed with satd NaHC03 (3 x 50 mL), H20 (3 x 50 mL), brine (3 x 50 mL) , dried (MgS04) and the solvent evaporated in vacuo. Flash column chromatography (2% MeOH/98% CHC13) gave the product as a white foam (0.114 g, 50%): [a]D26 = +97.5° (c = 0.2, CHC13) ; 'H NMR (CDC13) 6 7.40-7.15 (m, 12H), 6.74 (s, 2H), 5.68-5.60 (m, 2H), 4.45-4.24 (m, 6H), 4.05-3.78 (m, 8H), 3.76-3.38 (m, 6H), 3.12-3.07 (m, 2H), 2.97-2.87 (m, 2H), 2.31-1.66 (m, 12H), 1.57-1.55 (m, 2H); MS (ES) m/z (relative intensity) 979 (M+> +Na, 100), 957 (M+> +1, 35); IR (neat) 3298, 2945, 1704, 1602, 1514, 1432, 1270 cm' l,l'-[[Pentane-l,5-diyl]dioxy]bis[ (HS,llaS)-10-[N-(2-phenylsulfonylethyloxycarbonyl) ] -11 -hydroxy- 7-methoxy-l,2,3,10,ll,lla-hexahydro~5H-pyrrolo[2,l-c][1,4]benzodiazepine-5-one] (100) The alcohol 98 (0.80 g, 0.78 mmol) was reacted (Method A) with DAIB (0.553 g, 1.72 mmol) and TEMPO (0.024 g, 0.16 ramol) in CH2C12 (5 mL). The product was obtained (flash column chromatography 2% MeOH/98% CHC13) as a white foam (0.641 g, 80%). [a]D26 +73.0° (c = 1.0, CHC13) ; aH NMR (CDC13) 5 8.12-7.40 (m, 10H) , 7.20 (s, 2H), 6.97-6.75 (m, 2H), 5.70-5.55 (m, 2H), 4.86 (bs, 2H), 4.81 (bs, 1H), 4.43 (bs, 1H), 4.22-3.97 (m, 6H), 3.88 (s, 6H), 3.75-3.16 (m, 11H), 2.21-1.82 (mf 12H), 1.72-1.54 (m, 2H); MS (ES) m/z (relative intensity) 1044 (M+> +Na, 60), 1043 (100), 1021 (MT-, 23); IR (neat) 3345, 2953, 1748, 1628, 1319 cm'1. Deprotection of groups in compounds 66 and 67 1,1'-I[Pentane-l,5-diyl]dioxy]bis[ (115fllaS) -10-[N- (4- (L-glut amyl ca rbonyl oxy) ben zyl oxyca rbonyl) ]-ll -hydroxy - 7 -methoxy-1,2,3,10,11,lla-hexahydro-5H-pyrrolo[2,l-c][1,4]benzodiazepine-5-one] (68) (Figure Removed) A solution of the bis-diallyl ester 66 (0.69 g, 0.49 mmol, 1 eq), Pd(PPh3)4 (0.08 g, 0.069 mmol, 0.14 eq) and morpholine (0.33 g, 0.33 mL, 3.8 mmol, 7.7 eq) in anhydrous CH2C12 (12 mL) was stirred at room temperature for 18 h. The solvent was decanted and the solid residue was washed with EtOAc (2 x 15 mL) , CH2C12 (2 x 15 mL) , dissolved in MeOH (5 mL) and passed down an IRC 50, weakly acidic, ion exchange column, eluting with MeOH (100 mL) . The solvent was removed in vacua to give the product as a white foam (0.48 g, 78%) . [a] 16-2 +98.9° (c = 0.273, MeOH); aH NMR (CD3OD) 6 7.21 (m, 4H) , 7.07 (m, 6H), 6.72 (s, 2H) , 5.66 (d, J= 9 Hz, 2H) , 5,24 (d, J= 12.2 Hz, 2H), 4.22 (m, 2H) , 3.93 (m, 6H) , 3.84 (m, 8H) , 3.62 (m, 4H), 3.45 (m, 4H) , 2.43 (m, 4H) , 2.21 (m, 2H) , 2.15-1.95 (m, 10H), 1.8 (m, 4H) , 1.6 (m, 2H) . l,l'-[[Pentane-l,5-diyl]dioxy]bis[ (llS,llaS)-10-[N-(4-(diprop-2-enyl-L-glutamylcarbonylamino)benzyloxycarbonyl) ]-ll-hydroxy-7-methoxy-l,2f3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c] [l,4]benzodiazepine-5-one] (69) (Figure Removed)A solution of the bis-diallyl ester 67 (0.29 g, 0.21 mmol, 1 eq), Pd(PPh3)« (0.033 g, 0.028 mmol, 0.14 eq) and morpholine (0.14 g, 0.14 ml, 1.6 mmol, 7.7 eq) in anhydrous CH2C12 (5 mL) was stirred at room temperature for 18 h. The solvent was decanted and the solid residue was washed with EtOAc (2 x 15 mL), CH2C12 (2 x 15 mL), dissolved in MeOH (5 mL) and passed down an IRC 50, weakly acidic, ion exchange column, eluting with MeOH (100 mL). The solvent was removed in vacuo to give the product as an off-white solid (0.2 g 77%). [a]D15'6 +143.2° (c = 0.234, MeOH); aH NMR (CD3OD) 67.33 (d, J= 6.4 Hz, 6H), 7.15 (m, 6H), 6.66 (s, 2H), 5.65 (d, J= 9.4 Hz, 2H), 5.26 (d, J- 11.44 Hz, 2H), 4.76 (d, J = 11.8 Hz,, 2H), 4.36 (m, 2H), 4.15-3.8 (m, 12H), 3.65 (m, 4H), 2.39 (m, 4H), 2.25-1.9 (m, 12H), 1.8 (m, 4H), 1.55 (m, 2H). Deprotection of N-10 protected dimers to give 1,1-[(pentane-1,5-diyl)dioxy]bis[(llaS)-7-methoxy-l,2,3,lla-tetrahydro-5H-pyrrolo[2,l-c][1,43benzodiazepine-5-one] (61) (Figure Removed) Deprotection of 1,l'-[[Pentane-1,5-diyl]dioxy]bis[ (IIS,llaS)-10-(allyloxycarbonyl)-11-hydroxy-7-methoxy-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c][l,4]benzodiazepine-5-one] (60) A solution of the alloc protected dimer (0.068 g, 0.09 mmol, 1 eq) , Pd(PPh3)4 (0.005 g, 0.08 mrnol, 0.2 eq) and pyrrolidine (0.016 g, 0.018 mL, 0.22 mmol, 2.5 eq) in anhydrous CH2C12 (5 mL) was stirred at room temperature for 4 h and the solvent removed in vacuo. The product was purified by flash column chromatography (5% MeOH/95% CHC13) to give 61 as a yellow solid (0.04 g, 82%). *H NMR (CDC13) 6 7.61 (d, J= 4.4 Hz, 2H) , 7.44 (s, 2H), 6.73 (s, 2H), 4.04 (m, 4H), 3.87 (s, 6H), 3.8 (m, 2H), 3.69 (m, 2H), 3.5 (m, 2H), 2.24 (m, 4H), 2.05-1.85 (m, 8H), 1.6 (m, 2H). Deprotection of l,l'-[[Pentane-1,5-diyl]dioxy]bis[ (HS,llaS) -10-[N- (4- (L-glutamylcarbonyloxy)benzyloxycarbonyl) ]-ll-hydroxy-7-methoxy-l,2,3,10,ll,lla-hexahydro-5H-pyrrolo[2,l-c] [1/4]benzodiazepine-5-one] (68) An aliquot of a 10 mM stock solution of the carbamate dimer prodrug was diluted to 100 uM in Carboxypeptidase G2 (CPG2) assay buffer (100 mM Tris-HCl, pH 7.3; 260 uM ZnCl2) . CPG2 (1 unit) was added and the reaction was incubated at 37°C. The reaction was monitored by reversed phase HPLC (C18 column, 5 uM particle size, 250 mm x 4.6 mm; mobile phase 70% H20/30% CH3CN/0.1% TFA; detection at 245 nM) with 20 uL aliquots being injected at 0, 15, 30, 45, 60 and 75 min. The conversion of the prodrug to the parent inline 61 was complete in 75 min. Synthesis of alternate monomer isocyanate Methyl-2-isocyanotobenzoate (71) (Figure Removed) Triphosgene (5.8 g, 19.8 MMol) was added to a solution of methyl anthranilate (70) (1 g, 6.6 iranol) and pyridine (8.0 mL, 99 iranol) in CH2C12 and stirred at room temperature for 4 hours. The reaction mixture was washed in IN HCl (3 x 50 mL), H20 (3 x 50 mL), brine (3 x 50 mL) and dried over MgS04. Excess solvent was removed in vacua to give a quantitative yield of pure material (71). aH NMR (250 MHz, CDC13) 8 8.02 (dd, 1H, J= 1.7, 7.9 Hz, aromatic H), 7.46 (dt, 1H, J= 1.5, 5.8, 7.7 Hz, aromatic H) , 7.29-7.25 (m, 1H, H-3), 7.12 (dd, 1H, J = 1.2, 8.03 Hz, H-2), 3.96 (s, 3H, OMe); IR (neat) v 3676, 3483, 3369, 2956, 2844, 2592, 2306, 2191, 2129, 1944, 1735, 1697, 1611, 1540, 1457, 1320, 1280, 1170, 1097, 1045, 968, 879, 831, 768, 704 cm"1; MS m/z 178 9 (M+>+l), 153, 152, 146, 121, 120, 92, 90. Synthesis of alternate monomer carbamate Methyl-2- ((E)-3-pheyl-allyloxycarbonylamino)-benzoate (72) (Figure Removed) A solution of methyl-2-isocyanatobenzoate (71) (1 g, 5.6 iranol) in CH2C12 (100 mL) was cooled to 0°C and treated with pyridine (7 mL, 84 mmol). Cinnamyl alcohol (2.25 g, 1.68 mmol) in CH2C12 (150 mL) was added dropwise to the stirring solution over a period of 1 hour and the reaction mixture was allowed to stir for 12 hours, under nitrogen. The reaction mixture was washed in brine (100 mL) . The aqueous phase was washed with CH2C12 (3 x 100 mL) . The organic phases were combined and washed with IN HCl (200 mL) and dried over MgS04. Purification was achieved through flash column chromatography (80% Pet ether:EtOAc) to furnish the carbamate (72) as a colourless solid (85%). :H NMR (250 MHz, CDC13) 8 10.9 (br, s, 1H, NH), 8.45 (dd. 1H, J= 1.0, 8.5 Hz, aromatic H), 8.01 (dd, 1H, J= 1.66, 8.0 Hz, aromatic H) , 7.54 (ddd, 1H, J= 1.67, 7.3, 8.5 Hz, aromatic H), 7.5-7.2 (m, 5H, coc aromatic H), 7.03 (ddd, 1H, J= 1.2, 8.0 Hz, aromatic H), 6.71 (d, 1H, J - 16 Hz, 3'-H), 6.35 (dt, 1H, J= 6.3, 16 Hz, 2'-H), 4.48 (dd, 2H, J= 1.3, 6.3, I'-H), 3.91 (s, 1H); 13C NMR (62.9 MHz, CDC13) 5 169.0 (ester carbonyl), 153.5 (CO - carbamate), 141.9 (aromatic quat), 136.8 (aromatic quat), 135.0 (methine), 131.3 (methine), 129.0 (methine), 128.4 (methine), 127.1 (methine), 123.9 (alkenic methine), 122.0 (methine), 119.3 (methine). 114.8 (aromatic quat), 66.1 (methylene - Coc), 57.7 (OCH3) . IR (neat) v 3295, 3116, 3025, 2957, 2242, 1956, 1927, 1814, 1732, 1697, 1592, 1532, 1450, 1305, 1245, 1165, 1090, 1038, 976, 913, 856, 836, 819, 766, 743, 692 cm"1; MS m/z 311 (M"-+l) , 268, 212, 188, 152, 117. Synthesis of protected PBD 2-((E)-3-phenyl-aJ.lyloxycarbonylamino)-benzoic acid (73) (Figure Removed) Compound 72 (7.989 g, 26 mmol) was dissolved in aqueous methanol (3:1, 1200 mL) with THF (2-3 drops) to aid solubility. A solution of LiOH (3.12 g, 130 mmol) was added as a solid to the stirring reaction mixture at 0°C and stirred for 40 minutes. The reaction was allowed to return to room temperature and stirred for 16 hours, at which time TLC (70% Pet etherrEtOAc) revealed complete reaction. Excess methanol and THF were evaporated in vacuo and the remaining solution acidified to pH 2 with cone. HC1, to furnish a colourless precipitate (73), which was collected by filtration and dried (92%) . aH NMR (250 MHz, CDC13) 5 10.85 (s, IE, acid OH), 10.80 (s, 1H, carbamate NH), 8.34-8.32 (d, 1H, aromatic H), 8.00 (dd, 1H, J= 1.5, 7.9 Hz, aromatic H), 7.70-7.55 (m, 1H, aromatic H) , 7.53-7.10 (m, 5H, aromatic coc), 6.75 (d, 1H, J = 16 Hz, 3'-H), 6.46 (dt, 1H, J= 6.2, 16 Hz, 2'-H), 4.82 (d, 2H, J = 6.3 Hz, I'-H); I3C NMR (62.9 MHz, CDC13) 8 170.6 (ester carbonyl), 153.2 (CO carbamate), 142.0 (aromatic guat), 137.1 (aromatic quat), 135.1 (methine), 134.5 (alkenic methine), 132.5 (methine), 131.0 (methine), 127.7 (methine), 125.0 (alkenic methine), 122.6 (methine), 119.0 (methine), 116.5 (aromatic quat), 66.5 (methylene Coc). MS m/z 297 (M+'+l), 265, 252, 236, 224, 205, 189, 176, 149, 138, 117. {2-[1- (3-hydroxymethyl-pyrrolidin-l-yl) -methanoyl]-phenyl} carbamicacid (E)-3-phenyl-allyl ester (74) (Figure Removed) A solution of DCC (4.54 g, 22 mmol) dissolved in the minimum of CH2Cl2 was added dropwise to compound 73 (6 g, 0.020 mol) in at -5°C, whilst stirring, with a catalytic amount of DMF (10 drops). The reaction mixture was stirred for a further five minutes before HOBt (3.0 g, 22 mmol) in CH2C12 was added. The reaction mixture was allowed to stir for 1 hour at -5°C, then left to stir for 12 hours at room temperature. The colourless precipitate of DCU was removed by filtration and the remaining reaction mixture cooled to -5°C. (S)-pyrrolidinemethanol (2.37 mL, 24 mmol) was added and the reaction mixture allowed to warm to room temperature and, stirred for 12 hours. TLC (70% Pet ether/EtOAc) revealed reaction completion. The reaction mixture was washed with NaHC03 (4 x 100 mL) , NH (IIS, llaS) -ll-hydroxy-10- ( (E) -3-phenyl-allyloxycarbonyl) hexahydro-5H-pyrrolo[2,l-c][l,4]benzodiazepin-5-one (75) (Figure Removed) A solution of DMSO (4.6 mL 65 mmol) in CHZC12 was added over 20 minutes to a stirring solution of oxalyl chloride (16.5 mL, 33 mmol) in CH'2C12 at -40°C and left to stir for a further 20 minutes. Compound 74 (7 g, 18 mmol) was dissolved in CH2Cl2 and added over a period of 45 minutes to the reaction mixture. Once addition was complete, the reaction mixture was stirred at -40°C for a further 60 minutes. Over a period of 30 minutes TEA (10.8 mL, 77 mmol) in CH2C12 was added and stirred for a further 30 minutes, and then warmed to room temperature. The reaction mixture was washed with IN HCl (3 x 100 mL), H20 (3 x 100 mL), brine (3 x 100 mL) and dried over MgSO<. purification was achieved via flash column chromatography and excess solvent removed in vacuo to yield compound as a solid colourless powder> +256° (c = 1, CHC13); aH NMR (250 MHz, CDC13) 5 7.76 (dd, 1H, J = 1.6, 7.5 Hz, aromatic H), 7.60-7.30 (m, 5H, Coc aromatic H), 6.46 (d, 1H, J = 16 Hz, 3'-H}, 6.30-5.94 (m, 1H, 2'-H), 5.71 (d, 1H, J= 9.7 Hz, H-ll), 4.86-4.72 (m, 2H, I'-H), 3.77-3.41 (m, 4H, lla-H, H-3), 2.17-1.78 (m, 4H, 1-H, H-2); 13C NMR (62.9MHz, DMSO) 5 163.9 (C=0 amide), 155.8 (C=0 carbinolamine), 136.7 (aromatic quat), 135.0 (aromatic quat), 131.8 (methine), 131.4 (methine), 129.5 (methine), 129.1 (methine), 128.8 (methine), 127.3 (methine), 86.0 (methine CC-11), 67.1 (rnethylene coc), 61.2 (methine C-lla), 46.8 (methylene C-3), 29.1 (raethylene C-l), 23.5 (methylene C-2); MS m/z 379 (M+'+l), 363, 317, 277, 245, 225, 117. PBD Deprotection (Figure Removed) A solution of Pd(PPh3) WE CLAIM: 1. A method of synthesis of protected pyrrolobenzodiazepine compound of formula (I): (Formula Removed) comprising the steps of: (a) reacting a compound of formula (II) with a compound of formula (III) to yield a compound of formula (IV): (Formula Removed) (b) converting the compound of formula (IV) to a compound of formula (I)having the sub-steps of: (iii) hydrolysis of the ester -C(=0)OR12; (iv) coupling of resulting acid with a compound of formula: wherein M is (Formula Removed) either OH or (c) when M is OH, oxidation of the resulting compound to yield a compound of formula (la) ; (d) when M is - , palladium mediated unmasking of the resulting compound to yield a compound of formula (la); and (iii) converting the compound of formula (la) to a compound of formula (I)by: (a) direct etherification of compound of formula (la) or treatment of compound of formula (la) with R1:LSH and preferably Lewis Acid or treatment of compound of formula (la) with R1:LNH and preferably Lewis Acid, depending upon whether X is 0, S or NH, respectively. (Formula Removed) wherein the dotted lines indicate the optional presence of a double bond between C1 and C2 or C2 and C3; R2 and R3 are independently selected from -H, -OH, =O, =CH2, -CN, -R, OR, =CH-R, O-SO2-R, CO2R and COR; R6, R7 and R9 are independently H, R, OH, OR, SH, SR, NH2, NHR, NRR' , nitro, MesSn and halo; R8 is either selected from H, R, OH, OR, SH, SR, NH2, NHR, NRR' , nitro, Me3Sn and halo or the compound is a dimer with each monomer being the same of different and being of the relevant formula, where the R8 groups of each monomer form together a bridge having the formula -X-R"-X-, where R" is a C3-12 alkylene group, which chain may be interrupted by one or more heteroatoms and/or aromatic rings, and each X is independently selected from 0, S and NH; R10-OC(=O)- forms a nitrogen protecting group wherein R10 is such as herein described; R11 is either H or R; R12 is an optionally substituted C1-4 alkyl group; R and R' are independently selected from optionally substituted C1-20 alkyl, C3-20 heterocyclyl, and C5-20 aryl; wherein T is 0 or S, and each Rac is independently selected from C1-10 alkyl or together can be a C1-3 alkylene group. 2. A method as claimed in claim 1, wherein Rlc is an optionally substituted C1-30 alkyl group, C3-30 heterocyclyl group or a C5-30 aryl group or a divalent version of one of these groups linked to another moiety. |
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1149-DELNP-2006-Abstract-(23-12-2008).pdf
1149-DELNP-2006-Claims-(23-12-2008).pdf
1149-DELNP-2006-Claims-(31-03-2009).pdf
1149-DELNP-2006-Correspondence-Others-(03-03-2009).pdf
1149-DELNP-2006-Correspondence-Others-(12-02-2009).pdf
1149-DELNP-2006-Correspondence-Others-(13-02-2009).pdf
1149-DELNP-2006-Correspondence-Others-(17-03-2011).pdf
1149-DELNP-2006-Correspondence-Others-(23-12-2008).pdf
1149-DELNP-2006-Correspondence-Others.pdf
1149-DELNP-2006-Description (Complete)-(23-12-2008).pdf
1149-DELNP-2006-Description (Complete)-(31-03-2009).pdf
1149-delnp-2006-description (complete).pdf
1149-DELNP-2006-Form-1-(23-12-2008).pdf
1149-delnp-2006-form-13(25.02.2009).pdf
1149-DELNP-2006-Form-2-(12-02-2009).pdf
1149-DELNP-2006-Form-2-(23-12-2008).pdf
1149-DELNP-2006-Form-27-(17-03-2011).pdf
1149-DELNP-2006-Form-3-(23-12-2008).pdf
1149-DELNP-2006-Petition-137-(13-02-2009).pdf
| Patent Number | 233948 | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Indian Patent Application Number | 1149/DELNP/2006 | |||||||||
| PG Journal Number | 21/2005 | |||||||||
| Publication Date | 22-May-2009 | |||||||||
| Grant Date | 22-Apr-2009 | |||||||||
| Date of Filing | 03-Mar-2006 | |||||||||
| Name of Patentee | SpiroGen Limited | |||||||||
| Applicant Address | 79 GEORGE STREET, RYDE, HAMPSHIRE PO33 2JF, UK | |||||||||
Inventors:
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| PCT International Classification Number | C07D 487/04 | |||||||||
| PCT International Application Number | PCT/GB2004/003873 | |||||||||
| PCT International Filing date | 2004-09-10 | |||||||||
PCT Conventions:
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