Inhibitors of post-proline cleaving proteases

Abstract

Novel compounds that are inhibitors of one or most post-proline cleaving proteases, e.g. dipeptidyl peptidase IV, according to general formula (1). R1 is H or CN, X1 is O, S, CH2, CHF, CF2, CH(CH3), C(CH3)2 or CH(CN), and b is 1 or 2. G1 is H or a group according to the formula —CH2—X2—(CH2)a-G3 and G2 is H or a group according to the formula —CH2—(CH29a-G3, provided that one of G1 and G2 is H and the other is not H. X2 is O, S, or CH2, and a is 0, 1 or 2, provided that when a is 1 then X2 is CH2. G3 is a group according to one of general formulae 2-4, where the variables have meaning given in the description. The compounds are useful in the treatment of i.a. type 2 diabetes and impaired glucose tolerance.

Description

The present invention relates to novel compounds that are inhibitors of post-proline aminopeptidases. The compounds are useful as antiproliferative agents and in the treatment of, inter alia, type 2 diabetes and impaired glucose tolerance.

BACKGROUND

The enzyme dipeptidyl peptidase IV, herein abbreviated DP-IV (and elsewhere as DAP-IV or DPP-IV) and also known by the classification EC.3.4.14.5, is a serine protease that cleaves the N-terminal dipeptide from peptides that begin with the sequence H-Xaa-Pro (where Xaa is any amino acid, although preferably a lipophilic one, and Pro is proline). It will also accept as substrates peptides that begin with the sequence H-Xaa-Ala (where Ala is alanine). DP-IV was first identified as a membrane-bound protein. More recently a soluble form has been identified.

Initial interest in DP-IV focussed on its role in the activation of T lymphocytes. DP-IV is identical to the T cell protein CD26. It was proposed that inhibitors of DP-IV would be capable of modulating T cell responsiveness, and so could be developed as novel immunomodulators. It was further suggested that CD26 was a necessary co-receptor for HIV, and thus that DP-IV inhibitors could be useful in the treatment of AIDS.

Attention was given to the role of DP-IV outside the immune system. It was recognised that DP-IV has a key role in the degradation of several peptide hormones, including growth hormone releasing hormone (GHRH) and glucagon-like peptide-1 and -2 (GLP-1 and GLP-2). Since GLP-1 is known to have a potentiating effect on the action of insulin in the control of post-prandial blood glucose levels it is clear that DP-IV inhibitors might also be usefully employed in the treatment of type II diabetes and impaired glucose tolerance. At least two DP-IV inhibitors are currently undergoing clinical trials to explore this possibility.

Several groups have disclosed inhibitors of DP-IV. While some leads have been found from random screening programs, the majority of the work in this field has been directed towards the investigation of substrate analogues. Inhibitors of DP-IV that are substrate analogues are disclosed in, for example, U.S. Pat. No. 5,462,928, U.S. Pat. No. 5,543,396, WO95/15309 (equivalent to U.S. Pat. No. 5,939,560 and EP 0731789), WO98/19998 (equivalent to U.S. Pat. No. 6,011,155), WO99/46272 and WO99/61431.

More recently a number of proteins have been found that share some of the enzymatic properties of DP-IV. Some, such as FAP and DPP-8, have sequence homology with DP-IV, while others, such as QPP, have no such homology but nevertheless mimic the aminodipeptidase activity of DP-IV. The physiological function of these newer proteases is still being investigated. FAP has been implicated in invasive processes such as cancer metastasis and endometriosis, and QPP appears to be involved in immune-cell apoptosis. It is also possible that some of these proteases share a common function. This redundancy would allow continuing normal physiological function in the event of a failure in the expression or function of one of the proteases.

In order to further define the roles of these newer proteases it is important to have the tools to manipulate selectively each one or the whole class. Therefore there exists a need for specific and potent inhibitors of each of these proteases, and also for potent non-specific inhibitors of the class of post-proline cleaving aminodipeptidases.

SUMMARY OF THE INVENTION

We disclose herein a series of novel compounds that are inhibitors of one or more post-proline cleaving proteases, and specifically compounds according to general formula 1.

In general formula 1, R¹ is H or CN, X¹ is O, S, CH₂, CHF, CF₂, CH(CH₃), C(CH₃)₂ or CH(CN), and b is 1 or 2. G¹ is H or a group according to the formula —CH₂—X²—(CH₂)_a-G³ and G² is H or a group according to the formula —CH₂ (CH₂)_a-G³, provided that one of G¹ and G² is H and the other is not H. X² is O, S or CH₂, and a is 0, 1 or 2, provided that when a is 1 then X² is CH₂. G³ is a group according to one of general formulae 2-4.

X³, X⁴ and X⁵ are either nitrogen N or CH, provided that at least two of X³, X⁴ and X⁵ are N. X⁶ is either O or NH. R² is either H or alkyl. R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷ and R⁸ are selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN. X⁷ is CH₂, O, S or NH. R⁹ is either H or alkyl. R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN. R¹⁵ and R¹⁶ are each independently H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl or CH₂-L-R⁷, where L is a covalent bond, CH═CH, C≡C or —C₆H₄—, and R¹⁷ is H, alkyl or aryl, or R¹⁵ and R¹⁶ together are a group according to one of general formulae 5-7.

R¹⁸ is H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl or N(alkyl)₂, and R¹⁹ is H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl or N(alkyl)₂. The integers d and e are 0, 1, 2 or 3 such that d+e is 3, 4 or 5, and f is 1, 2 or 3. When R¹⁵ and R¹⁶ are both H then X¹ may not be S or CH₂ if b is 1.

Preferred compositions are inhibitors of non-membrane associated post-proline cleaving proteases. The most preferred compositions are selective for non-membrane associated proteases (e.g. for example inhibitors of one or more of QPP, DPP-8 and/or DPP-9).

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention relates to a series of novel α-amino acyl derivatives of saturated nitrogen-containing heterocycles according to general formula 1.

In general formula 1, the group R¹ is either a hydrogen atom H or a nitrile group CN. The group X¹ is selected from an oxygen atom 0, a sulphur atom S, a methylene group CH₂, a monofluoromethylene group CHF, a difluoromethylene group CF₂, an ethylidene group CH(CH₃), a 2-propylidene group C(CH₃)₂ and a cyanomethylene group CH(CN). The integer b is either 1 or 2, such that the nitrogen-containing ring has 5 or 6 members.

The group G¹ is either H or a group according to the formula —CH₂—X²—(CH₂)_a-G³ and the group G² is either H or a group according to the formula —CH₂—(CH₂)_a-G³, provided that one of G¹ and G² is H and the other is not H. The group X² is selected from O, S and CH₂. The integer a is 0, 1 or 2, provided that when a is 1 then X² is CH₂.

The group G³ is selected from a group according to general formula 2, a group according to general formula 3 and a group according to general formula 4.

In general formula 2, the groups X³, X⁴ and X⁵ are selected from nitrogen N and methine CH, provided that at least two of X³, X⁴ and X⁵ are nitrogen. Preferably X³, X⁴ and X⁵ are all nitrogen. The group X⁶ is selected from O and NH. R² is selected from H and alkyl. R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷ and R⁸ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.

In general formula 3, the group X⁷ is selected from CH₂, O, S and NH. R⁹ is selected from H and alkyl. R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.

In general formula 4, R¹⁵ and R¹⁶ are each independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and CH₂-L-R⁷, where L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄— and R¹⁷ is selected from H, alkyl and aryl, or R¹⁵ and R¹⁶ together are a group selected from general formula 5, general formula 6 and general formula 7.

In these general formulae, the group R¹⁸ is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂, and the group R¹⁹ is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. The integers d and e are selected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5, and the integer f is selected from 1, 2 and 3.

When R¹⁵ and R¹⁶ are both H then X¹ may not be S or CH₂ if b is 1.

The term alkyl, as used herein, denotes saturated hydrocarbon groups with between 1 and 10 carbon atoms, including straight-chain, branched and mono- and polycycloalkyl groups, such as methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, cyclopentyl, cyclohexylmethyl, 2-cyclohexyl-2-propyl, bicyclo[2.2.2]octyl and the like.

The term alkenyl, as used herein, denotes monounsaturated hydrocarbon groups with between 2 and 10 carbon atoms, including straight-chain, branched and mono- and polycycloalkenyl groups, such as vinyl, allyl, methallyl, cyclohex-3-enyl and the like.

The term aryl, as used herein, denotes monocyclic and fused bicyclic aromatic groups, including carbocyclic groups, such as phenyl and naphthyl, and heteroaryl groups with up to three heteroatoms selected from nitrogen, oxygen and sulphur, such as pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, isothiazolyl, pyridyl, pyrimidinyl, indolyl, quinolinyl and the like. Unless otherwise specified, aryl groups may optionally be substituted with up to three groups independently selected from alkyl, OH, O-alkyl, Cl, F, Br, NH₂, NH-alkyl, N(alkyl)₂, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂, NO₂ and CN.

The term aralkyl, as used herein, denotes alkyl groups that are substituted by, or fused to, one or more aryl groups, including benzyl, phenethyl, indanyl, fluorenyl and the like.

The term acyl, as used herein, denotes a group selected from H—CO, alkyl-CO, aryl-CO and aralkyl-CO, including formyl, acetyl, benzoyl, phenylacetyl and the like.

The term polyfluoroalkyl, as used herein, denotes an alkyl group wherein all the hydrogen atoms on one or more of the carbon atoms are replaced by fluorine atoms, including trifluoromethyl, 2,2,2-trifluoroethyl and the like.

In one preferred embodiment of the invention R¹ is H.

In another preferred embodiment of the invention R¹ is CN.

In another preferred embodiment of the invention X¹ is CH₂.

In another preferred embodiment of the invention X¹ is S.

In another preferred embodiment of the invention b is 1.

In another preferred embodiment of the invention b is 2.

In another preferred embodiment of the invention a is 0.

In another preferred embodiment of the invention a is 0 and X² is CH₂.

In another preferred embodiment of the invention a is 1.

In another preferred embodiment of the invention a is 1 and X² is CH₂.

In another preferred embodiment of the invention a is 2 and X² is CH₂.

In another preferred embodiment of the invention the compound is a compound according to general formula 8.

In another preferred embodiment of the invention the compound is a compound according to general formula 9.

In another preferred embodiment of the invention the compound is a compound according to general formula 10.

In another preferred embodiment of the invention the compound is a compound according to general formula 11.

In another preferred embodiment of the invention the compound is a compound according to general formula 12.

In another preferred embodiment of the invention the compound is a compound according to general formula 13.

It will be recognised that certain of the compounds within the scope of the present invention are capable of forming salts with suitable acids or bases. To the extent that such salts are pharmaceutically acceptable they are included within the scope of this invention.

It will further be recognised that certain of the compounds within the scope of the present invention are capable of existing as optical isomers, such as enantiomers and diastereomers. All such optical isomers and mixtures thereof, including but not limited to racemates, are included within the scope of the invention.

The compounds of the present invention are inhibitors of post-proline cleaving proteases such as DPP-IV, QPP, FAP, DPP-8 (DPRP-1) and DPP-9 (DPRP-2). As such they may be useful in the treatment of diseases in which dysregulation of these enzymes or their endogenous substrates plays a role or the disease is ameliorated by inhibition of such enzymes. Accordingly, in further aspects, the present invention provides for the use of compounds according to the present invention in the preparation of pharmaceutical compositions, and for the use of such compositions a therapeutic agents.

Preferred compositions which are inhibitors for QPP may have G²=H, b=1 or 2 and/or a=0 or 1. Further preferred compositions having b=2 include G1 groups having a=0 or 1 and X² is CH₂ Further preferred compositions having b=2 have X¹═CH₂ or S, for example Example 38 of Table 2. Further preferred compositions having b=1 include G1 groups having a=0 or 1 and X² is CH₂. Further preferred compositions having b=1 have X¹═S or CH₂ or CF₂, for example, Example 42 of Table 2.

The compounds of the present invention can be prepared by methods generally known in the art and illustrated in the following non-limiting examples.

EXAMPLES

Example 1

(2S)-1-[N^ω,N^ω-(Dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide

N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysine (5 g, 10.7 mmol) was dissolved in CH₂Cl₂ (100 mL). The solution was cooled to 0° C., L-prolinamide (1.78 g, 11.7 mmol) and PYBOP® (6.7 g, 12.8 mmol) were added, and the pH adjusted to pH9 with triethylamine. After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). The solution was washed with 0.3M KHSO₄ (2×50 mL), sat. NaHCO₃ (2×50 mL), water (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (N^α-(tert-butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide (4.05 g, 7.2 mmol, 67%).

B. (2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide (3.95 g, 7.02 mmol) was dissolved in dry THF (100 mL). The solution was cooled to 0° C., triethylamine (1.4 g, 14 mmol) was added followed by the slow addition of trifluoroacetic anhydride (2.97 g, 14.1 mmol). The pH was adjusted to pH9 with triethylamine. After 30 min the reaction mixture was diluted with ethyl acetate (100 mL), washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography on silica gel (eluant: 60% pet ether, 40% ethyl acetate) to give a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (3.3 g, 6.11 mmol, 87%).

C. (2S)-1-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-pyrrolidine-2-carbonitrile (3.1 g, 5.7 mmol) was dissolved in THF (80 mL). Diethylamine (20 mL) was added. After 2 h at room temperature the solvent was removed in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (1.63 g, 5.03 mmol, 89%).

D. (2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (100 mg, 0.31 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (170 mg, 1.18 mmol). After 30 mins sodium triacetoxyborohydride (330 mg, 1.56 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (38 mg, 0.068 mmol, 11%). Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-N^ω-(cinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.073 mmol, 12%).

E. (2S)-1-[N^ω,N^ω-(Dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.057 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^ω,N^ω-(dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride (37 mg, 0.053 mmol, 93%).

[M+H]⁺=457.3

¹H NMR (CD₃OD): δ 1.35-1.55 (2H, m), 1.75-2.00 (2H, m), 2.05-2.23 (6H, m), 3.10-3.29 (4H, m), 3.61-3.68 (2H, m), 4.00-4.03 (4H, m), 4.20-4.30 (1H, m), 4.82-4.93 (1H, m), 6.34-6.39 (2H, m), 6.94 (2H, d, J=5.8 Hz), 7.31-7.37 (6H, m), 7.39-7.53 (4H, m) ppm.

Example 2

(2S)-1-[N^ω-(Cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-1-[N^ω-(Cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(cinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.057 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^α-(cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride (37 mg, 0.053 mmol, 93%).

[M+H]⁺=341.5

¹H NMR (CD₃OD): δ 1.29-1.55 (2H, m), 1.72-1.80 (2H, m), 1.90-2.11 (2H, m), 2.16-2.29 (6H, m), 3.02-3.09 (2H, m), 3.65-3.69 (2H, m), 3.78-3.82 (2H, m), 4.23-4.27 (1H, m), 4.81-4.82 (1H, m), 4.91-4.99 (1H, m), 6.21-6.32 (1H, m), 6.86 (1H, d, J=6.1 Hz), 7.26-7.35 (3H, m), 7.37-7.40 (2H, m) ppm.

Example 3

(2S)-1-[N^ω,N^ω-(Dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-(N^α-(tert-Butyloxycarbonyl)-L-ornithyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-L-ornithyl)pyrrolidine-2-carbonitrile was prepared by the method described for the lysine derivative in Example 1.

B. (2S)-(N-(tert-Butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (200 mg, 0.65 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (180 mg, 1.25 mmol). After 30 mins sodium triacetoxyborohydride (343 mg, 1.63 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-ornithinyl)-pyrrolidine-2-carbonitrile (77 mg, 0.14 mmol, 22%). Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as (2S)-1-(N^α-(tert-butyloxycarbonyl)-N^ω-(cinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (78 mg, 0.18 mmol, 28%).

C. (2S)-1-[N^ω,N^ω-(Dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (67 mg, 0.12 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^ω,N^ω-(dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride (82 mg, 0.12 mmol, 100%).

[M+H]⁺=443.3

¹H NMR (CD₃OD): δ 1.98-2.12 (4H, m), 2.22-2.29 (4H, m), 3.27-3.31 (4H, m), 3.62-3.67 (2H, m), 3.96 (4H, d, J=7.5 Hz), 4.30-4.40 (1H, m), 4.80-4.83 (1H, m), 6.34-6.41 (2H, m), 6.96 (2H, d, J=15.6 Hz), 7.31-7.39 (6H, m), 7.49-7.53 (4H, m) ppm.

Example 4

(2S)-1-[N^ω-(Cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-1-[N^ω-(Cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(cinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (71 mg, 0.17 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^ω-(cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride (91 mg, 0.16 mmol, 100%).

[M+H]⁺=327.5

¹H NMR (CD₃OD): δ 1.70-1.88 (2H, m), 1.97-2.01 (2H, m), 2.14-2.32 (4H, m), 3.08-3.13 (2H, m), 3.29-3.31 (3H, m), 3.68-3.71 (2H, m), 3.79-3.82 (2H, m), 4.29-4.31 (1H, m), 4.87-4.91 (1H, m), 6.29-6.31 (1H, m), 6.86 (1H, d, J=15.8 Hz), 7.29-7.30 (3H, m), 7.44-7.48 (2H, m) ppm.

Example 5

3-[N^ω,N^ω-(Dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-thiazolidine

N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysine (2.73 g, 6 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.53 g, 10 mmol), water-soluble carbodiimide (1.34 g, 7 mmol), thiazolidine (1.28 g, 18 mmol) and N-methylmorpholine (1.0 g, 10 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet ether) to give a white solid identified as 3-[N^α-(tert-butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiazolidine (2.55 g, 4.85 mmol, 81%).

B. 3-[N^α-(tert-Butyloxycarbonyl)-L-lysinyl]thiazolidine

3-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiazolidine (1.15 g, 2.13 mmol) was dissolved in acetonitrile (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-[N^α-(tert-butyloxycarbonyl)-L-lysinyl]thiazolidine (530 mg, 1.67 mmol, 78%).

C. 3-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-lysinyl)thiazolidine

3-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (200 mg, 0.6 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (400 mg, 3.0 mmol). After 30 mins sodium triacetoxyborohydride (534 mg, 2.54 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as 3-(N^α-(tert-butyloxycarbonyl)-N^ω,N^ω-(dicinnamyl)-L-lysinyl)thiazolidine (139 mg, 0.25 mmol, 40%).

D. 3-[N^ω,N^ω-(Dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride

3-(N^α-(tert-Butyloxycarbonyl)-N^ω, N^ω-(di-cinnamyl)-L-lysinyl)thiazolidine (139 mg, 0.25 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^ω,N^ω-(dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride (127 mg, 0.24 mmol, 96%).

[M+H]⁺=450.2

¹H NMR (CD₃OD): δ 1.49-1.55 (2H, m), 1.89-1.98 (4H, m), 3.01-3.30 (4H, m), 3.4-3.5 (4H, m), 3.7-3.9 (3H, m), 4.0-4.2 (3H, m), 4.2-4.8 (2H, br m), 6.38-6.44 (2H, m), 6.99-6.93 (2H, m), 7.34-7.37 (5H, m), 7.51-7.60 (4H, m) ppm.

Example 6

3-[N^ω,N^ω-(Cinnamyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(cinnamyl)-L-lysinyl)thiazolidine

3-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (200 mg, 0.6 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (400 mg, 3.0 mmol). After 30 mins sodium triacetoxyborohydride (534 mg, 2.54 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% triethylamine, 5% methanol, 94% chloroform) to give a colourless oil identified as 3-(N^α-(tert-butyloxycarbonyl)-N^ω, N^ω-(cinnamyl)-L-lysinyl)thiazolidine (215 mg, 0.50 mmol, 83%).

B. 3-[N^ω,N^ω-(Cinnamyl)-L-lysinyl]thiazolidine dihydrochloride

3-(N^α-(tert-Butyloxycarbonyl)-N^ω,N^ω-(cinnamyl)-L-lysinyl)thiazolidine (215 mg, 0.5 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^ω,N^ω-(cinnamyl)-L-lysinyl]thiazolidine dihydrochloride (160 mg, 0.40 mmol, 79%).

[M+H]⁺=334.4

¹H NMR (CD₃OD): δ 1.28-1.30 (1H, m), 1.51-1.53 (1H, m), 1.79-1.78 (1H, m), 1.93-1.98 (2H, m), 2.9-3.3 (5H, m), 3.6-3.8 (5H, m), 4.30-4.70 (5H, m), 6.2-6.3 (1H, m), 6.85-6.91 (1H, m), 7.1-7.7 (5H, m) ppm.

Example 7

1-[N^ω-(Cyclohexylmethyl)-L-ornithinyl]pyrrolidine dihydrochloride

A. 1-[N^ω-(Benzyloxycarbonyl)-N^α-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine

N^ω-(Benzyloxycarbonyl)-N^α-(tert-butyloxycarbonyl)-L-ornithine (5.49 g, 15 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (3.37 g, 22 mmol), water-soluble carbodiimide (3.46 g, 18 mmol), pyrrolidine (1.28 g, 18 mmol) and N-methylmorpholine (2.0 g, 20 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). The solution was washed with 0.3M KHSO₄ (2×50 mL), sat. NaHCO₃ (2×50 mL), water (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 90% ethyl acetate, 10% pet. ether) to give a colourless oil identified as 1-[N^ω-(benzyloxycarbonyl)-N^α-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine (5.15 g, 12.3 mmol, 82%).

B. 1-[N^α-(tert-Butyloxycarbonyl)-L-ornithinyl]pyrrolidine

1-[N^ω-(Benzyloxycarbonyl)-N^α-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine (2.15 g, 5.13 mmol) was dissolved in methanol (80 mL). This solution was hydrogenated over 10% Pd/C (400 mg). After 2 h the catalyst was filtered off and washed with methanol (50 mL). The combined filtrates were evaporated in vacuo to give an off white solid identified as 1-[N^α-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine (1.35 g, 4.74 mmol, 94%).

C. 1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(cyclohexylmethyl)-L-ornithinyl)pyrrolidine

1-[N^α-(tert-Butyloxycarbonyl)-L-ornithinyl]pyrrolidine (100 mg, 0.35 mmol) was dissolved in methanol (25 mL). To this solution was added cyclohexanecarboxaldehyde (44 mg, 0.39 mmol). After 30 mins sodium triacetoxyborohydride (148 mg, 0.70 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% triethylamine, 5% methanol, 94% chloroform) to give a colourless oil identified as 1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(cyclohexylmethyl)-L-ornithinyl)pyrrolidine (51 mg, 0.18 mmol, 52%).

D. 1-[N^ω-(Cyclohexylmethyl)-L-ornithinyl]pyrrolidine dihydrochloride

1-(N^α-(tert-Butyloxycarbonyl)-N^ω-(cyclohexylmethyl)-L-ornithinyl)pyrrolidine (215 mg, 0.5 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N^ω-(cyclohexylmethyl)-L-ornithinyl]pyrrolidine dihydrochloride (160 mg, 0.40 mmol, 79%).

[M+H]⁺=282.3

¹H NMR (CD₃OD): δ 0.93-1.24 (3H, m), 1.66-1.81 (15H, m), 2.50-2.70 (2H, m), 2.71-2.88 (2H, m), 3.2-3.48 (6H, m), 4.08 (1H, m), 8.35-8.38 (1H, m), 8.80-8.85 (1H, m) ppm.

Example 8

3-[N^ω-Me-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride

A. N^α-(tert-Butyloxycarbonyl-N^ω-benzyl-L-lysine methyl ester

N^α-(tert-Butyloxycarbonyl-L-lysine methyl ester (6.1 g, 22.2 mmol) was dissolved in methanol (100 mL). To this solution was added benzaldehyde (1.9 g, 17.5 mmol). After 2 hours sodium triacetoxyborohydride (5.8 g, 27.3 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 5% methanol, 94% chloroform) to give a colourless oil identified as N^α-(tert-butyloxycarbonyl-N^ω-benzyl-L-lysine methyl ester (5.2 g, 14.2 mmol, 82%).

B. N^α-tert-Butyloxycarbonyl-N^ω-benzyl-N^ω-methyl-L-lysine methyl ester

N^α-tert-Butyloxycarbonyl-N^ω-benzyl-L-lysine methyl ester (5.0 g, 14.2 mmol) was dissolved in methanol (100 mL). To this solution was added formaldehyde (37% solution in water, 10 mL). After 2 hours sodium triacetoxyborohydride (3.9 g, 18.4 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat. Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as N^α-tert-butyloxycarbonyl-N^ω-benzyl-N^ω-methyl-L-lysine methyl ester (5.2 g, 14.2 mmol, 100%).

C. N^α-tert-Butyloxycarbonyl-N^ω-methyl-L-lysine methyl ester

N^α-tert-Butyloxycarbonyl-N^ω-benzyl-N^ω-methyl-L-lysine methyl ester (5.0 g, 14.2 mmol) was dissolved in methanol/water (9:1, 100 mL). To this solution was added ammonium formate (1.6, 19.3 mmol) and 10% palladium on charcoal (2 g). After 3 hours at 60° C. the catalyst was filtered off through celite and the residue washed with methanol (50 mL). The combined filtrates were evaporated in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as N^α-(tert-butyloxycarbonyl-N^ω-methyl-L-lysine methyl ester (3.48 g, 12.5 mmol, 93%).

D. N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine methyl ester

N^α-tert-Butyloxycarbonyl-N^ω-methyl-L-lysine methyl ester (3.1 g, 11.1 mmol) was dissolved in dichloromethane (100 mL). To this solution was added 1,1-dimethyl-2,2,2-trichloroethyl chloroformate (3.0 g, 12.5 mmol) and triethylamine (2.3 g, 23 mmol). After 18 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). This solution was washed with 0.3M KHSO₄ (1×50 mL), sat NaHCO₃ (1×50 mL), water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil purified by flash chromatography on silica gel (eluant: 30% ethyl acetate, 70% pet. ether) to give colourless oil identified as N^α-(tert-butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine methyl ester (3.28 g, 6.98 mmol, 63%).

E. N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine

N^α-(tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine methyl ester (3.1 g, 6.6 mmol) was dissolved in tetrahydrofuran (100 mL). 1M Lithium hydroxide (7 mL, 7.0 mmol) was added. After 3 hours at room temperature the reaction mixture was diluted with ethyl acetate (150 mL), washed with 1M HCl (1×50 mL), water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give colourless oil identified as N^α-(tert-butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine (2.94 g, 6.45 mmol, 98%).

F. 3-(N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysinyl)thiazolidine

N^α-(tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysine (700 mg, 1.51 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (410 mg, 3.0 mmol), water-soluble carbodiimide (250 mg, 1.3 mmol), thiazolidine (170 mg, 1.9 mmol) and N-methylmorpholine (1.0 g, 10 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×25 mL), sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether) to give a white solid identified as 3-(N^α-tert-butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysinyl)thiazolidine (758 mg, 1.42 mmol, 94%).

G. 3-(N^α-tert-Butyloxycarbonyl-N^ω-methyl-L-lysinyl)thiazolidine

3-(N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^ω-methyl-L-lysinyl)thiazolidine (730 mg, 1.36 mmol) was dissolved in acetic acid (30 mL). Zinc powder (200 mg) was added. After stirring at room temperature for 18 hours the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). The solution was washed with sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-(N^α-tert-butyloxycarbonyl-N^ω-methyl-L-lysinyl)thiazolidine (438 mg, 1.32 mmol, 97%).

H. 3-[N^α-tert-Butyloxycarbonyl-N^ω-methyl-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine

3-(N^α-tert-Butyloxycarbonyl-N^ω-methyl-L-lysinyl)thiazolidine (50 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (20 mL). To this solution was added 2-naphtaldehyde (26 mg, 0.17 mmol). After 2 hours sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 4% methanol, 96% chloroform) to give a colourless oil identified as 3-[N^α-tert-butyloxycarbonyl-N^ω-methyl-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine (51 mg, 0.11 mmol, 72%).

I. 3-[N^ω-Methyl-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride

3-[N^α-tert-Butyloxycarbonyl-N^ω-methyl-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine (44 mg, 0.093 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^ω-methyl-N^ω-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride (37 mg, 0.083 mmol, 89%).

[M+H]⁺=372.2

¹H NMR (CD₃OD): δ 1.50-1.53 (2H, m), 1.91-1.98 (4H, m), 2.82 (3H, s), 3.08-3.19 (4H, m), 3.36-3.75 (5H, m), 4.32-4.47 (2H, m), 4.60-4.71 (2H, m), 7.55-7.59 (2H, m), 7.65-7.68 (1H, m), 7.90-8.00 (3H, m), 8.10-8.12 (1H, m) ppm.

Example 9

3-[N^ω-Methyl-N^ω-(1-Napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride

A. 3-[N-(tert-Butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine

N-(tert-Butyloxycarbonyl)-O^ω-methyl-L-glutamic acid (6.28 g, 24 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 ml). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (5.5 g, 36 mmol), water-soluble carbodiimide (5.38 g, 28 mmol), thiazolidine (2.48 g, 28 mmol) and N-methylmorpholine (3.0 g, 30 mmol). The mixture was stirred for 18 h at 0° C. to room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 ml). The solution was washed with 0.3M KHSO₄ (2×30 ml), sat. NaHCO₃ (2×30 ml), water (2×30 ml) and brine (1×30 ml), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether 60-80) to give a brown oil identified as 3-[N-(tert-butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine (4.0 g, 12 mmol, 50%).

B. 3-[N,N-Di-(tert-butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine

3-[N-(tert-Butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine (3.2 g, 9.6 mmol) was dissolved in acetonitrile (20 mL). Di-tert-butyl dicarbonate (3.14 g, 14.4 mmol) and 4-dimethylaminopyridine (235 mg, 1.93 mmol) were added. After 18 hours at room temperature further di-tert-butyl dicarbonate (3.14 g, 14.4 mmol) was added. After a further 3 days at room temperature the solvent was evaporated in vacuo the residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether 60-80) to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine (2.0 g, 4.63 mmol, 48%).

C. 3-[N,N-Di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine

3-[N,N-di-(tert-butyloxycarbonyl)-O^ω-methyl-L-glutamyl]thiazolidine (950 mg, 2.22 mmol) was dissolved in THF (50 ml). 1M Lithium hydroxide (5.5 ml, 5.5 mmol) was added. The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 ml). The solution was washed with 0.3M KHSO₄ (2×20 ml), water (2×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine (912 mg, 2.2 mmol, 98%).

D. 3-[2-(N,N-Di-(tert-butyloxycarbonyl)amino)-5-hydroxypentanoyl]thiazolidine

3-[N,N-Di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine (912 mg, 2.2 mmol) was dissolved in tetrahydrofuran (30 mL). This solution was cooled to −20° C., N-methylmorpholine (300 mg, 2.96 mmol) and isobutyl chloroformate (387 mg, 2.83 mmol) were added. After 20 mins at −20° C. the reaction mixture was added to a solution of sodium borohydride (182 mg, 4.8 mmol) in water (5 mL) at 0° C. After 1 hour the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-[2-(N,N-di-(tert-butyloxycarbonyl)amino)-5-hydroxy-pentanoyl]thiazolidine (800 mg, 2.0 mmol, 92%).

E. 3-[2-(N,N-Di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine

3-[2-N,N-((Di-tert-butyloxycarbonyl)amino)-5-hydroxypentanoyl]thiazolidine (800 mg, 2.0 mmol) was dissolved in dichloromethane (50 mL). Dess-Martin periodinane (933 mg, 2.2 mmol) was added. After 1 hour at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. Purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether 60-80) to give a colourless oil identified as 3-[2-(N,N-di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine (210 mg, 0.52 mmol, 26%).

F. 3-[N,N-Di-(tert-butyloxycarbonyl-N^ω-methyl-N^ω-(1-napthylmethyl)-L-ornithyl]-thiazolidine

3-[N,N-Di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine was dissolved in 1,2-dichloroethane (20 mL). To this solution was added N-methyl-1-napthylmethylamine. After 2 hours sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl-N^ω-methyl-N^ω-(1-napthylmethyl)-L-ornithyl]thiazolidine.

G. 3-[N^ω-Methyl-N^ω-(1-Napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride

3-[N,N-Di-(tert-butyloxycarbonyl-N^ω-methyl-N^ω-(1-napthylmethyl)-L-ornithyl]thiazolidine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^ω-Me,N^ω-(1-napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride.

Example 10

3,3-Difluoro-1-[N^ω-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride

A. 1-(tert-Butyloxycarbonyl)-3-pyrrolidone

(3R)-1-(tert-Butyloxycarbonyl)-3-hydroxypyrrolidine (980 mg, 5.3 mmol) was dissolved in CH₂Cl₂ (40 ml). Dess-Martin periodinane (2.5 g, 5.8 mmol) was added. The mixture was stirred for 3 hours at room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (300 ml). The solution was washed with sat. NaHCO₃, water and brine, dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. The residue was purified by flash chromatography on silica gel (eluant: 20% ethyl acetate, 80% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3-pyrrolidone (842 mg, 4.6 mmol, 87%).

B. 1-(tert-Butyloxycarbonyl)-3,3-difluoropyrrolidine

1-(tert-Butyloxycarbonyl)-3-pyrrolidone (810 mg, 4.4 mmol) was dissolved in CH₂Cl₂ (30 ml). (Diethylamino)sulphur trifluoride (2.2 g, 13.7 mmol) was added to this solution at 0° C. The mixture was stirred for 18 hours at 0° C. to room temperature then carefully poured into sat. NaHCO₃ (100 ml). The mixture was stirred for 15 min then extracted with CH₂Cl₂. The organic extract was washed with water and brine, dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography (eluant: 10% ethyl acetate, 90% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3,3-difluoropyrrolidine (580 mg, 2.8 mmol, 64%).

C. 3,3-Difluoropyrrolidine hydrochloride

1-(tert-Butyloxycarbonyl)-3,3-difluoropyrrolidine (540 mg, 2.6 mmol) was dissolved in 4M HCl/dioxan (30 ml). The solution was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give an off white solid identified as 3,3-difluoropyrrolidine hydrochloride (370 mg, 2.6 mmol, 100%).

D. 1-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine

N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysine (1.14 g, 2.4 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 ml). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (394 mg, 2.9 mmol), water-soluble carbodiimide (680 mg, 3.4 mmol), 3,3-difluoropyrrolidine hydrochloride (380 mg, 2.43 mmol) and N-methylmorpholine (400 mg, 4 mmol). The mixture was stirred for 18 h at 0° C. to room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 ml). The solution was washed with 0.3M KHSO₄, sat. NaHCO₃, water and brine, dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 65% ethyl acetate, 35% pet. ether 60-80) to give a white solid identified as 1-[N^α-(tert-butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine (1.0 g, 1.8 mmol, 75%).

E. 1-[N^α-(tert-Butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine

1-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoro-pyrrolidine (1.01 g, 1.8 mmol) was dissolved in THF (20 ml). Diethylamine (5 ml) was added. The mixture was stirred for 3 hours at room temperature then the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N^α-(tert-butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine (598 mg, 1.78 mmol, 99%).

F. 1-[N^α-(tert-Butyloxycarbonyl)-N^ω-(2-methylbutyl)-L-lysinyl]-3,3-difluoro-pyrrolidine

1-[N^α-(tert-Butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine was dissolved in 1,2-dichloroethane (20 mL). To this solution was added 2-methylbutanal. After 2 hours sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 1-[N^α-(tert-butyloxycarbonyl)-N^ω-(2-methylbutyl)-L-lysinyl]-3,3-difluoropyrrolidine.

G. 3,3-Difluoro-1-[N^ω-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride

1-[N^α-(tert-Butyloxycarbonyl)-N^ω-(2-methylbutyl)-L-lysinyl]-3,3-difluoropyrrolidine was dissolved in 4M HCl/dioxan (20 ml). The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give a colourless oil identified as 3,3-difluoro-1-[N^ω-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride.

Example 11

1-[N^ω-(3-Cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride

A. 3-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine

N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysine (2.5 g, 5.34 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.44 g, 10.6 mmol), water-soluble carbodiimide (1.35 g, 6.5 mmol), thiomorpholine (710 mg, 6.9 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 3-[N^α-(tert-butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine (2.70 g, 4.88 mmol, 91%).

B. 3-[N^α-(tert-Butyloxycarbonyl)-L-lysinyl]thiomorpholine

3-[N^α-(tert-Butyloxycarbonyl)-N^ω-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine (2.6 g, 4.7 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-[N^α-(tert-butyloxycarbonyl)-L-lysinyl]thiomorpholine (1.2 g, 3.637 mmol, 77%).

C. 3-[N^α-(tert-Butyloxycarbonyl)-N^ω-(3-cyclohexenylmethyl)-L-lysinyl]-thiomorpholine

3-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)thiomorpholine (150 mg, 0.45 mmol) was dissolved in methanol (25 mL). To this solution was added 3-cyclohexanecarboxaldehyde (400 mg, 0.45 mmol). After 30 mins sodium triacetoxyborohydride (150 mg, 0.71 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 3-(N^α-(tert-butyloxycarbonyl)-N^ω-(3-cyclohexenylmethyl)-L-lysinyl)thiomorpholine (66 mg, 0.12 mmol, 26%).

D. 1-[N^ω-(3-Cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride

3-(N^α-(tert-Butyloxycarbonyl)-N^ω-(3-cyclohexenylmethyl)-L-lysinyl)thiomorpholine (66 mg, 0.12 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N^ω-(3-cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride (62 mg, 0.12 mmol, 100%).

[M+H]⁺=326.2

Example 12

(2S)-1-[N^ω-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]thiazolidine dihydrochloride

A. 3-[N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine

N^α-(tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithine (2.5 g, 5.9 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 30 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.6 g, 11.9 mmol), water-soluble carbodiimide (1.4 g, 7.6 mmol), thiazolidine (650 mg, 7.3 mmol) and N-methylmorpholine (2.0 g, 20 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×25 mL), sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether) to give a colourless oil identified as 3-[N^α-tert-butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine (758 mg, 1.42 mmol, 94%).

B. 3-(N^α-tert-Butyloxycarbonyl-L-ornithinyl)thiazolidine

3-[N^α-tert-Butyloxycarbonyl-N^ω-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine (130 mg, 0.26 mmol) was dissolved in acetic acid (30 mL). Zinc powder (100 mg) was added. After stirring at room temperature for 18 hours the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). The solution was washed with sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-(N^α-tert-butyloxycarbonyl-L-ornithinyl)thiazolidine (80 mg, 0.26 mmol, 100%).

C. 3-[N^α-tert-Butyloxycarbonyl-N^ω-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]thiazolidine

3-(N^α-tert-Butyloxycarbonyl-L-ornithinyl)thiazolidine (80 mg, 0.26 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (80 mg, 0.6 mmol), water-soluble carbodiimide (65 mg, 0.32 mmol), niflumic acid (82 mg, 0.29 mmol) and N-methylmorpholine (100 mg, 1.0 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3-[N^α-tert-butyloxycarbonyl-N^ω-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]-thiazolidine (60 mg, 0.12 mmol, 45%).

D. (2S)-1-[N^ω-(2-(3′-Trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]-thiazolidine dihydrochloride

3-[N^α-tert-Butyloxycarbonyl-N^ω-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]thiazolidine (54 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as (2S)-1-[N^ω-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]thiazolidine dihydrochloride (47 mg, 0.10 mmol, 100%).

[M+H]⁺=468.0

¹H NMR (CD₃OD): δ 1.77-1.82 (2H, m), 1.84-2.00 (2H, m), 3.03-3.15 (4H, m), 3.41-3.51 (2H, m), 3.65-3.71 (2H, m), 3.80-3.87 (1H, m), 4.46-4.49 (2H, m), 4.65-4.72 (2H, m), 7.06-7.11 (1H, m), 7.61-7.11 (3H, m), 7.95 (1H, s), 8.09 (1H, d, J=4.7 Hz), 8.49 (1H, d, J=4.2 Hz) ppm.

Example 13

3,3-Difluoro-1-[N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride

A. 1-[N^α-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine

1-[N^α-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine was prepared as described for the lysine derivative in Example 9.

B. 3-Chloroanilinonicotinic acid

3-Chloroaniline was dissolved in xylene. 2-Aminonicotinic acid was added. The reaction mixture was heated at 150° C. for 18 hours after which time the reaction mixture was diluted with ethyl acetate giving an off-white solid identified as 3-chloroanilinonicotinic acid.

C. 3,3-Difluoro-[N^α-tert-butyloxycarbonyl-N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithinyl]pyrrolidine

1-[N^α-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate, water-soluble carbodiimide, 3-chloroanilinonicotinic acid and N-methylmorpholine. After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3,3-difluoro-[N^α-tert-butyloxycarbonyl-N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)]-L-ornithinyl)pyrrolidine.

D. 3,3-Difluoro-1-[N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride

3,3-Difluoro-[N^α-tert-butyloxycarbonyl-N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)]-L-ornithinyl)pyrrolidine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3,3-difluoro-1-[N^ω-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride.

Example 14

3-[N^ω-6-Chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

A. 4,6-Dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine

Cyanuric chloride (1.844 g, 10 mmol) was dissolved in acetonitrile (20 mL). The solution was cooled to −20° C. A solution of 2,5-dichloroaniline (1.62 g, 10 mmol) and triethylamine (1.0 g, 10 mmol) was slowly added. After 1 hour at −20° C. the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). The solution was washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was recrystallised from ethyl acetate/hexane to give an off white solid identified as 4,6-dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine (1.86 mg, 6.0 mmol, 60%).

B. 3-[N^α-tert-Butyloxycarbonyl-N^ω-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine

3-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (800 mg, 2.58 mmol) was dissolved in dichloromethane (30 mL). To this solution was added 4,6-dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine (810 mg, 2.6 mmol) and triethylamine (300 mg, 3.0 mmol). After 2 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). This solution was washed with water (2×30 mL) and brine (1×30 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 3-[N^α-tert-butyloxycarbonyl-N^ω-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine (1.33 g, 2.23 mmol, 86%).

C. 3-[N^ω-6-Chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

3-[N^α-tert-Butyloxycarbonyl-N^ω-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine (59 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^ω-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride (55 mg, 0.098 mmol, 98%).

[M+H]⁺=492.2, 494.4

¹H NMR (CD₃OD): δ 1.46-1.51 (2H, m), 1.65-1.67 (2H, m), 1.80-1.96 (2H, m), 3.05-3.14 (2H, m), 3.38-3.42 (2H, m), 3.55-3.75 (4H, m), 4.31-4.36 (2H, m0, 4.40-4.52 (1H, m), 4.63-4.95 (2H, m), 7.15-7.18 (1H, m), 7.40-7.45 (1H, m), 8.15-8.25 (1H, m) ppm.

Example 15

3-[N^ω-4-(2′,5′-Dichloroanilino)-6-hydroxy-1,3,5-triazinyl)-L-lysinyl]thiazolidine bis(trifluoroacetate)

A. 3-[N^ω-4-(2′,5′-Dichloroanilino)-6-hydroxy-1,3,5-triazinyl)-L-lysinyl]thiazolidine bis(trifluoroacetate)

3-[N^α-tert-Butyloxycarbonyl-N^ω-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)]-L-ornithinyl)thiazolidine (54 mg, 0.09 mmol) was dissolved in trifluoroacetic acid (20 mL) and water (2 mL). After 2 hours at 70° C. the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^ω-4-(2′,5′-dichloroanilino)-6-hydroxy-1,3,5-triazinyl)-L-lysinyl]thiazolidine bis(trifluoroacetate) (63 mg, 0.089 mmol, 97%).

[M+H]⁺=472.1, 474.2

¹H NMR (CD₃OD): δ 1.42-1.47 (2H, m), 1.62-1.67 (2H, m), 1.82-1.89 (2H, m), 3.04-3.16 (4H, m), 3.70-3.75 (2H, m), 3.84-3.91 (1H, m), 4.25-4.32 (2H, m), 4.45-4.54 (2H, m), 4.64-4.70 (2H, m), 7.05-7.15 (1H, m), 7.34-7.38 (1H, m), 7.49-7.55 (1H, m), 7.80-7.92 (1H, m) ppm.

Example 16

3-[N^ω-4-(2′,5′-Dichloroanilino)-6-methylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-[N^α-tert-Butyloxycarbonyl-N^ω-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine

3-[N^α-tert-Butyloxycarbonyl-N^ω-3-chloro-5-(2′,5′-dichloroanilino)-2,4,6-triazinyl)]-L-ornithinyl)thiazolidine (120 mg, 0.20 mmol) was dissolved in 1M dimethylamine in tetrahydrofuran (25 mL). After 18 hours at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether) to give a white solid identified as 3-[N^α-tert-butyloxycarbonyl-N^ω-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine (110 mg, 0.18 mmol, 90%).

B. 3-[N^ω-4-(2′,5′-Dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]-thiazolidine dihydrochloride

3-[N^α-tert-Butyloxycarbonyl-N^ω-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine (110 mg, 0.18 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^ω-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride (105 mg, 0.18 mmol, 100%).

[M+H]⁺=499.1, 501.1

¹H NMR (CD₃OD): δ 1.52-1.55 (2H, m), 1.69-1.71 (2H, m), 1.90-1.98 (2H, m), 3.13-3.22 (8H, m), 3.48-3.62 (2H, m), 3.65-3.69 (4H, m), 4.37-4.39 (2H, m), 4.46-4.49 (1H, m), 4.57-4.77 (2H, m), 7.20-7.22 (1H, m), 7.45-7.50 (1H, m), 8.09-8.12 (1H, m) ppm.

The following compounds were prepared by analogous methods.

TABLE 1
Example No	n	X
17	3
1819	34
2021	34
2223	34
2425	34
2627	34

TABLE 2
Example No	n	X
28	2
293031	234
323334	234
353637	234
383940	234
41	2
424345	234
464748	234
49	2
505152	234

TABLE 3
Ex
No	a	b	X	R3	R4
53 54 55	111	343	S CH2	HHH
56	1	4		H
57	1	3	CF2	H
58	1	4		H
59	1	4	S	CH3
60	1	4		CH(CH3)2
61	1	4	CH2	CH3
62	1	4		CH(CH3)2
63	1	3	S	CH(CH3)2
64	1	3	CH2	CH(CH3)2
65	2	3	S	H
66	2	4		H
67	2	3	CH2	H
68	2	4		H
69 70 71 72 73 74 75 76	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
77	1	4	CH2	CH3
78	1	4		CH(CH3)2
79	1	3	S	CH(CH3)2
80	1	3	CH2	CH(CH3)2
81	2	3	S	H
82	2	4		H
83	2	3	CH2	H
84	2	4		H
85 86 87	111	343	S CH2	HHH
88	1	4		H
89	1	3	CF2	H
90	1	4		H
91	1	4	S	CH3
92	1	4		CH(CH3)2
93	1	4	CH2	CH3
94	1	4		CH(CH3)2
95	1	3	S	CH(CH3)2
96	1	3	CH2	CH(CH3)2
97	2	3	S	H
98	2	4		H
99	2	3	CH2	H
100	2	4		H
101102103104105106107108	11111111	34343444	S CH2 CF2S CH2	HHHHHCH3CH(CH3)2CH3
109	1	4		CH(CH3)2
110	1	3	S	CH(CH3)2
111	1	3	CH2	CH(CH3)2
112	2	3	S	H
113	2	4		H
114	2	3	CH2	H
115	2	4		H
116117118119	1111	3434	S CH2	HHHH
120	1	3	CF2	H
121	1	4		H
122	1	4	S	CH3
123	1	4		CH(CH3)2
124	1	4	CH2	CH3
125	1	4		CH(CH3)2
126	1	3	S	CH(CH3)2
127	1	3	CH2	CH(CH3)2
128	2	3	S	H
129	2	4		H
130	2	3	CH2	H
131	2	4		H
132133134135136137138139	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
140	1	4	CH2	CH3
141	1	4		CH(CH3)2
142	1	3	S	CH(CH3)2
143	1	3	CH2	CH(CH3)2
144	2	3	S	H
145	2	4		H
146	2	3	CH2	H
147	2	4		H
148149150151	1111	3443	S CH2CF2
152	1	4
153	1	4	S	CH3
154	1	4		CH(CH3)2
155	1	4	CH2	CH3
156	1	4		CH(CH3)2
157	1	3	S	CH(CH3)2
158	1	3	CH2	CH(CH3)2
159	2	3	S	H
160	2	4		H
161	2	3	CH2	H
162	2	4		H
163164165166167168169170	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
171	1	4	CH2	CH3
172	1	4		CH(CH3)2
173	1	3	S	CH(CH3)2
174	1	3	CH2	CH(CH3)2
175	2	3	S	H
176	2	4		H
177	2	3	CH2	H
178	2	4		H
179180181182183	11111	34343	S CH2 CF2	HHHHH
184	1	4		H
185	1	4	S	CH3
186	1	4		CH(CH3)2
187	1	4	CH2	CH3
188	1	4		CH(CH3)2
189	1	3	S	CH(CH3)2
190	1	3	CH2	CH(CH3)2
191	2	3	S	H
192	2	4		H
193	2	3	CH2	H
194	2	4		H
195196197198199200201202	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
203	1	4	CH2	CH3
204	1	4		CH(CH3)2
205	1	3	S	CH(CH3)2
206	1	3	CH2	CH(CH3)2
207	2	3	S	H
208	2	4		H
209	2	3	CH2	H
210	2	4		H
211212213214215	11111	34343	S CH2 CF2	HHHHH
216	1	4		H
217	1	4	S	CH3
218	1	4		CH(CH3)2
219	1	4	CH2	CH3
220	1	4		CH(CH3)2
221	1	3	S	CH(CH3)2
222	1	3	CH2	CH(CH3)2
223	2	3	S	H
224	2	3	CH2	H
225	2	4		H
226227228229230231232233	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
234	1	4	CH2	CH3
235	1	4		CH(CH3)2
236	1	3	S	CH(CH3)2
237	1	3	CH2	CH(CH3)2
238	2	3	S	H
239	2	4		H
240	2	3	CH2	H
241	2	4		H
242243244245246247	111111	343434	S CH2 CF2	HHHHHH
248	1	4	S	CH3
249	1	4		CH(CH3)2
250	1	4	CH2	CH3
251	1	4		CH(CH3)2
252	1	3	S	CH(CH3)2
253	1	3	CH2	CH(CH3)2
254	2	3	S	H
255	2	4		H
256	2	3	CH2	H
257	2	4		H
258259260261262263264265	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
266	1	4	CH2	CH3
267	1	4		CH(CH3)2
268	1	3	S	CH(CH3)2
269	1	3	CH2	CH(CH3)2
270	2	3	S	H
271	2	4		H
272	2	3	CH2	H
273	2	4		H
274275276277278279	111111	343434	S CH2 CF2	HHHHHH
280	1	4	S	CH3
281	1	4		CH(CH3)2
282	1	4	CH2	CH3
283	1	4		CH(CH3)2
284	1	3	S	CH(CH3)2
285	1	3	CH2	CH(CH3)2
286	2	3	S	H
287	2	4		H
288	2	3	CH2	H
289	2	4		H
290291292293294295296297	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
298	1	4	CH2	CH3
299	1	4		CH(CH3)2
300	1	3	S	CH(CH3)2
301	1	3	CH2	CH(CH3)2
302	2	3	S	H
303	2	4		H
304	2	3	CH2	H
305	2	4		H
306307308309310311	111111	343434	S CH2 CF2	HHHHHH
312	1	4	S	CH3
313	1	4		CH(CH3)2
314	1	4	CH2	CH3
315	1	4		CH(CH3)2
316	1	3	S	CH(CH3)2
317	1	3	CH2	CH(CH3)2
318	2	3	S	H
319	2	4		H
320	2	3	CH2	H
321	2	4		H
322323324325326327328329	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
330	1	4	CH2	CH3
331	1	4		CH(CH3)2
332	1	3	S	CH(CH3)2
333	1	3	CH2	CH(CH3)2
334	2	3	S	H
335	2	4		H
336	2	3	CH2	H
337	2	4		H
338339340341342343	111111	343434	S CH2 CF2	HHHHHH
344	1	4	S	CH3
345	1	4		CH(CH3)2
346	1	4	CH2	CH3
347	1	4		CH(CH3)2
348	1	3	S	CH(CH3)2
349	1	3	CH2	CH(CH3)2
350	2	3	S	H
351	2	4		H
352	2	3	CH2	H
353	2	4		H
354355356357358359360361	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
362	1	4	CH2	CH3
363	1	4		CH(CH3)2
364	1	3	S	CH(CH3)2
365	1	3	CH2	CH(CH3)2
366	2	3	S	H
367	2	4		H
368	2	3	CH2	H
369	2	4		H
370371372373374375	111111	343434	S CH2 CF2	HHHHHH
376	1	4	S	CH3
377	1	4		CH(CH3)2
378	1	4	CH2	CH3
379	1	4		CH(CH3)2
380	1	3	S	CH(CH3)2
381	1	3	CH2	CH(CH3)2
382	2	3	S	H
383	2	4		H
384	2	3	CH2	H
385	2	4		H
386387388389390391392393	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
394	1	4	CH2	CH3
395	1	4		CH(CH3)2
396	1	3	S	CH(CH3)2
397	1	3	CH2	CH(CH3)2
398	2	3	S	H
399	2	4		H
400	2	3	CH2	H
401	2	4		H
402403404405406407	111111	343434	S CH2 CF2	HHHHHH
408	1	4	S	CH3
409	1	4		CH(CH3)2
410	1	4	CH2	CH3
411	1	4		CH(CH3)2
412	1	3	S	CH(CH3)2
413	1	3	CH2	CH(CH3)2
414	2	3	S	H
415	2	4		H
416	2	3	CH2	H
417	2	4		H
418419420421422423424425	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
426	1	4	CH2	CH3
427	1	4		CH(CH3)2
428	1	3	S	CH(CH3)2
429	1	3	CH2	CH(CH3)2
450	2	3	S	H
451	2	4		H
452	2	3	CH2	H
453	2	4		H
454455456457458459	111111	343434	S CH2 CF2	HHHHHH
460	1	4	S	CH3
461	1	4		CH(CH3)2
462	1	4	CH2	CH3
463	1	4		CH(CH3)2
464	1	3	S	CH(CH3)2
465	1	3	CH2	CH(CH3)2
466	2	3	S	H
467	2	4		H
468	2	3	CH2	H
469	2	4		H
470471472473474475476477	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
478	1	4	CH2	CH3
479	1	4		CH(CH3)2
480	1	3	S	CH(CH3)2
481	1	3	CH2	CH(CH3)2
482	2	3	S	H
483	2	4		H
484	2	3	CH2	H
485	2	4		H
486487488489490491	111111	343434	S CH2 CF2	HHHHHH
492	1	4	S	CH3
493	1	4		CH(CH3)2
494	1	4	CH2	CH3
495	1	4		CH(CH3)2
496	1	3	S	CH(CH3)2
497	1	3	CH2	CH(CH3)2
498	2	3	S	H
499	2	4		H
500	2	3	CH2	H
501	2	4		H
502503504505506507508509	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
510	1	4	CH2	CH3
511	1	4		CH(CH3)2
512	1	3	S	CH(CH3)2
513	1	3	CH2	CH(CH3)2
514	2	3	S	H
515	2	4		H
516	2	3	CH2	H
517	2	4		H
518519520521522523524525	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
526	1	4	CH2	CH3
527	1	4		CH(CH3)2
528	1	3	S	CH(CH3)2
529	1	3	CH2	CH(CH3)2
530	2	3	S	H
531	2	4		H
532	2	3	CH2	H
533	2	4		H
534535536537538539540541	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
542	1	4	CH2	CH3
543	1	4		CH(CH3)2
544	1	3	S	CH(CH3)2
545	1	3	CH2	CH(CH3)2
546	2	3	S	H
547	2	4		H
548	2	3	CH2	H
549	2	4		H
550551552553554555556557	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
558	1	4	CH2	CH3
559	1	4		CH(CH3)2
560	1	3	S	CH(CH3)2
561	1	3	CH2	CH(CH3)2
562	2	3	S	H
563	2	4		H
564	2	3	CH2	H
565	2	4		H
566567568569570571572573	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
574	1	4	CH2	CH3
575	1	4		CH(CH3)2
576	1	3	S	CH(CH3)2
577	1	3	CH2	CH(CH3)2
578	2	3	S	H
579	2	4		H
580	2	3	CH2	H
581	2	4		H
582583584585586587588589	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
590	1	4	CH2	CH3
591	1	4		CH(CH3)2
592	1	3	S	CH(CH3)2
593	1	3	CH2	CH(CH3)2
594	2	3	S	H
595	2	4		H
596	2	3	CH2	H
597	2	4		H
598599600601602603604605	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
606	1	4	CH2	CH3
607	1	4		CH(CH3)2
608	1	3	S	CH(CH3)2
609	1	3	CH2	CH(CH3)2
610	2	3	S	H
611	2	4		H
612	2	3	CH2	H
613	2	4		H
614615616617618619620621	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
622	1	4	CH2	CH3
623	1	4		CH(CH3)2
624	1	3	S	CH(CH3)2
625	1	3	CH2	CH(CH3)2
626	2	3	S	H
627	2	4		H
628	2	3	CH2	H
629	2	4		H
630631632633634635636637	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
638	1	4	CH2	CH3
639	1	4		CH(CH3)2
640	1	3	S	CH(CH3)2
641	1	3	CH2	CH(CH3)2
642	2	3	S	H
643	2	4		H
644	2	3	CH2	H
645	2	4		H
646647648649650651652653	11111111	34343444	S CH2 CF2 S	HHHHHHCH(CH3)2CH3
654	1	4		CH(CH3)2	CH2
655	1	3	S	CH(CH3)2
656	1	3	CH2	CH(CH3)2
657	2	3	S	H
658	2	4		H
659	2	3	CH2	H
660	2	4		H
661662663664665666667668	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
669	1	4	CH2	CH3
670	1	4		CH(CH3)2
671	1	3	S	CH(CH3)2
672	1	3	CH2	CH(CH3)2
673	2	3	S	H
674	2	4		H
675	2	3	CH2	H
676	2	4		H
677678679680681682	111111	343444	SCH2CF2 S	HHHHCH3CH(CH3)2
683	1	4	CH2	CH3
684	1	4		CH(CH3)2
685	1	3	S	CH(CH3)2
686	1	3	CH2	CH(CH3)2
687	2	3	S	H
688	2	4	CH2	H
689690691692693694695696	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
697	1	4	CH2	CH3
698	1	4		CH(CH3)2
699	1	3	S	CH(CH3)2
700	1	3	CH2	CH(CH3)2
701	2	3	S	H
702	2	4		H
703	2	3	CH2	H
704	2	4		H
705706707708709	11111	34343	S CH2 CF2	HHHHH
710	1	4		H
711	1	4	S	CH3
712	1	4		CH(CH3)2
713	1	4	CH2	CH3
714	1	4		CH(CH3)2
715	1	3	S	CH(CH3)2
716	1	3	CH2	CH(CH3)2
717	2	3	S	H
718	2	4		H
719	2	3	CH2	H
720	2	4		H
721722723724725726727728	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
729	1	4	CH2	CH3
730	1	4		CH(CH3)2
731	1	3	S	CH(CH3)2
732	1	3	CH2	CH(CH3)2
733	2	3	S	H
734	2	4		H
735	2	3	CH2	H
736	2	4		H
737738739740741	11111	33434	SCH2 CF2	HHHHH
742	1	4	S	CH3
743	1	4		CH(CH3)2
744	1	4	CH2	CH3
745	1	4		CH(CH3)2
746	1	3	S	CH(CH3)2
747	1	3	CH2	CH(CH3)2
748	2	3	S	H
749	2	4		H
750	2	3	CH2	H
751	2	4		H
752753754755756757758759	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
760	1	4	CH2	CH3
761	1	4		CH(CH3)2
762	1	3	S	CH(CH3)2
763	1	3	CH2	CH(CH3)2
764	2	3	S	H
765	2	4		H
766	2	3	CH2	H
767	2	4		H
768769770771772773	11111	343434	S CH2 CF2	HHHHHH
774	1	4	S	CH3
775	1	4		CH(CH3)2
776	1	4	CH2	CH3
777	1	4		CH(CH3)2
778	1	3	S	CH(CH3)2
779	1	3	CH2	CH(CH3)2
780	2	3	S	H
781	2	4		H
782	2	3	CH2	H
783	2	4		H
784785786787788789790791	11111111	34343444	S CH2 CF2 S	HHHHHHCH3CH(CH3)2
792	1	4	CH2	CH3
793	1	4		CH(CH3)2
794	1	3	S	CH(CH3)2
795	1	3	CH2	CH(CH3)2
796	2	3	S	H
797	2	4		H
798	2	3	CH2	H
799	2	4		H

TABLE 4
Example
No	X	R
800801	SCH2
802803	SCH2
804805	SCH2
806807	SCH2
808809	SCH2
810811	SCH2
812813	SCH2
814815	SCH2
816817	SCH2
818819	SCH2
820821	SCH2
822823	SCH2
824825	SCH2
826827	SCH2
828829	SCH2
830831	SCH2
832833	SCH2
834835	SCH2
836837	SCH2
838839	SCH2
841842	SCH2
843844	SCH2
845846	SCH2
847	SCH2
848849	SCH2
850851	SCH2
852853	SCH2
854855	SCH2
856857	SCH2
858859	SCH2
860861	SCH2
862863	SCH2
864865	SCH2
866867	SCH2
868869	SCH2
870871	SCH2
872873	SCH2
874875	SCH2
876877	SCH2

TABLE 5
Example
No	n	X	R
878879880881	3434	S CH2
882883884885	3434	S CH2
886887888889	3434	S CH2
890891892893	3434	S CH2
894895896897	3434	S CH2
898899900901	3434	S CH2
902903904905	3434	S CH2
906907908909	3434	S CH2
910911912913	3434	S CH2
914915916917	3434	S CH2
9189199200	3434	S CH2
921922923924	3434	S CH2
925926927928	3434	S CH2
929930931932	3434	S CH2	Me
933934935936	3434	S CH2
937938939940	3434	S CH2
941942943944	3434	S CH2
945946	34	SCH2
947948949950	3434	S CH2
951952953954	3434	S CH2
955956957958	3434	S CH2
959960961962	3434	S CH2
963964965966	3434	S CH2
967968969970	3434	S CH2
971972973974	3434	S CH2
975976977978	4 34	S CH2
979980981982	3434	S CH2	MeS
983984985986	3434	S Ch2	MeO

TABLE 6
Example
No	n	X	R
987988989990	3434	S CH2
991992993994	4 34	S CH2
997998999998	3434	S CH2
999100010011002	3434	S CH2
1003100410051006	3434	S CH2
10071008 10091010	3434	S CH2
1011101210131014	3434	S CH2
1015101610171018	3434	S CH2
1019102010211022	3434	S CH2
1023102410251026	3434	S CH2
1027102810291030	3434	S CH2
1031103210331034	3434	S CH2
1035103610371038	3434	S CH2
1039104010411042	3434	S CH2	Me
1044104510461047	3434	S CH2
1048104910501051	3434	S CH2
1052105310541055	3434	S CH2
10561057	34	SCH2
1058105910601061	3434	S CH2
1062106310641065	3434	S CH2
1066106710681069	3434	S CH2
1070107110721073	3434	S CH2
1074107510761077	3434	S CH2
1078107910801081	3434	S CH2
1082108310841085	3434	S CH2
1086108710881089	3434	S CH2
1090109110921093	3434	S CH2	MeS
1094109510961097	3434	S CH2	MeO

TABLE 7
Example
No	n	X	R
1098109911001101	3434	S CH2
1102110311041105	3434	S CH2
1106110711081109	3434	S CH2
1110111111121113	3434	S CH2
1114111511161117	3434	S CH2
1118111911201121	3434	S CH2
1122112311241125	3434	S CH2
1125a112611271128	3434	S CH2
1129113011311132	3434	S CH2
1133113411351136	3434	S CH2
1137113811391140	3434	S CH2
1141114211431144	3434	S CH2
1145114611471148	3434	S CH2
1149115011511152	3434	S CH2
1153115411551156	3434	S CH2
11571158	34	SCH2
1159116011611162	3434	S CH2
1163116411651166	3434	S CH2
1167116811691170	3434	S CH2
1171117211731174	3434	S CH2
1175117611771178	3434	S CH2
1179118011811182	3434	S CH2
1183118411851186	3434	S CH2

TABLE 8
	Example
	No	n	X	R
	1187118811891190	3434	S CH2
	1191119211931194	3434	S CH2
	1195119611971198	3434	S CH2
	1199120012011202	3434	S CH2
	1203120412051206	3434	S CH2
	1207120812091210	3434	S CH2
	1211121212131214	3434	S CH2
	1215121612171218	3434	S CH2
	1219122012211222	3434	S CH2
	1223122412251226	3434	S CH2
	1227122812291230	3434	S CH2
	1231123212331235	3434	S CH2
	1235123612371238	3434	S CH2
	1239124012411242	3434	S CH2
	1243124412451246	3434	S CH2
	12471248	34	SCH2
	1249125012511252	3434	S CH2
	1253125412551256	3434	S CH2
	1257125812591260	3434	S CH2
	1261126212631264	3434	S CH2
	1265126612671268	3434	S CH2
	1269127012711272	3434	S CH2
	1273127412751276	3434	S CH2

Example 1277

1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride

A. 1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]thiomorpholine

1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (1.0 g, 2.27 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (461 mg, 3.41 mmol), water-soluble carbodiimide (521 mg, 2.72 mmol), thiomorpholine (281 mg, 2.72 mmol) and triethylamine (340 mg, 3.4 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]thiomorpholine (516 mg, 0.98 mmol, 43%).

B. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine

1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl thiomorpholine (500 mg, 0.95 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine (162 mg, 0.54 mmol, 56%).

C. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine

1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine (41 mg, 0.135 mmol) was dissolved in dichloroethane (10 mL). To this solution was added cyclohexanecarboxaldehyde (15 mg, 0.135 mmol). After 30 mins sodium triacetoxyborohydride (32 mg, 0.15 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine (25 mg, 0.063 mmol, 47%).

D. 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride

1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine (25 mg, 0.063 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride (23 mg, 0.063 mmol, 100%).

[M+H]⁺=300.3

Example 1278

1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride

A. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl thiomorpholine

1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine (41 mg, 0.135 mmol) was dissolved in 1,2-dichloroethane (10 mL). To this solution was added 2-quinolinecarboxaldehyde (32 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl thiomorpholine (32 mg, 0.072 mmol, 53%).

B. 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride

1-[2-(S)—N-(tert-Butyloxyarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine (12 mg, 0.027 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride (11.3 mg, 0.027 mmol, 100%).

[M+H]⁺=345.3

Example 1279

1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride

A. 1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine

1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (947 mg, 2.154 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (436 mg, 3.2 mmol), water-soluble carbodiimide (495 g, 2.58 mmol), piperidine (220 g, 2.58 mmol) and triethylamine (320 mg, 3.2 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine (556 mg, 1.1 mmol, 51%).

B. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine

1-[2-(S)—N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine (540 g, 1.1 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]piperidine (171 mg, 0.6 mmol, 57%).

C. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine

1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine (43 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (20 mL). To this solution was added cyclohexanecarboxaldehyde (17 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine (38 mg, 0.1 mmol, 66%).

D. 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride

1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine (38 mg, 0.1 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride (33 mg, 0.093 mmol, 93%).

[M+H]⁺=282.3

Example 1280

1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride

A. 1-[2-(S)—N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine

1-[2-(S)—N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine (24 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (25 mL). To this solution was added 2-quinolinecarboxaldehyde (24 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine (35 mg, 0.082 mmol, 55%).

B. 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride

1-[2-(S)—N-(tert-Butyloxyarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine (35 mg, 0.082 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride (26 mg, 0.065 mmol, 79%).

[M+H]⁺=327.3

Example 1281

3-Fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride

A. 1-(tert-Butyloxycarbonyl)-3-fluoropyrrolidine

N-(tert-Butyloxycarbonyl)-3-hydroxypyrrolidine (21.0 g, 10.7 mmol) was dissolved in CH₂Cl₂ (30 ml). (Diethylamino)sulphur trifluoride (1.72 g, 10.7 mmol) was added to this solution at −78° C. The mixture was stirred for 18 hours at −78° C. to room temperature then the reaction mixture was carefully poured into sat. NaHCO₃ (100 ml) and stirred for 15 min and extracted with CH₂Cl₂. The organic extract was washed with water and brine, dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography (eluant: 28% ethyl acetate, 72% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3-fluoropyrrolidine (1.14 g, 5.34 mmol, 50%).

B 3-Fluoropyrrolidine hydrochloride

1-(tert-Butyloxycarbonyl)-3-fluoropyrrolidine (1.14 g, 5.34 mmol) was dissolved in 4M HCl/dioxan (30 ml). The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give an off-white solid identified as 3-fluoropyrrolidine hydrochloride (640 mg, 5.2 mmol, 95%).

C. 3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine

1-[2-(S)—N-(tert-Butyloxyarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (950 mg, 2.15 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (395 mg, 2.6 mmol), water-soluble carbodiimide (572 mg, 3.0 mmol), 3-fluoropyrrolidine hydrochloride (270 g, 2.15 mmol) and triethylamine (320 mg, 3.2 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 3-fluoro1-[2-(S)—N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine (808 mg, 1.58 mmol, 73%).

D. 3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine

3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine (800 mg, 1.58 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine (316 mg, 1.04 mmol, 66%).

E. 3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine

3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine (150 mg, 0.52 mmol) was dissolved in methanol (20 mL). To this solution was added 3-cyclohexanecarboxaldehyde (63 mg, 0.57 mmol). After 30 mins sodium triacetoxyborohydride (220 mg, 1.04 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 3-fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine (176 mg, 0.46 mmol, 77%).

F. 3-Fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride

3-Fluoro-1-[2-(S)—N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine (176 mg, 0.46 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride (140 mg, 0.39 mmol, 963%).

[M+H]⁺=284.3

Example 1282

1-[2-(S)-Amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride

A. N-(tert-Butyloxycarbonyl)-L-homoserine lactone

L-Homoserine lactone 1.76 g, 12.8 mmol) was dissolved in DMF (30 mL). This solution was cooled to 0° C., triethylamine (1.41, 14.1 mmol) di-tert-butyl dicarbonate (3.35 g, 15.35 mmol) was added. After 18 hours at room temperature the solvent was evaporated in vacuo, the residue was taken up in dichloromethane (200 mL). This solution was washed with 1M KHSO₄ (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a white solid, recrystallised from EtOAc/pet ether to give a white solid identified as N-(tert-butyloxycarbonyl)L-homoserine lactone (2.25 mg, 11.2 mmol, 87%).

B. 1-[2-(S)—(N-(tert-Butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine

N-(tert-Butyloxycarbonyl)-L-homoserine lactone (100 mg, 0.5 mmol) was dissolved in tetrahydrofuran (30 mL). Piperidine (42 mg, 0.5 mmol) was added. After 72 hours at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil identified as 1-[2-(S)—(N-(tert-butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine (142 mg, 0.5 mmol, 100%).

C. 1-[2-(S)—(N-(tert-Butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine

1-[2-(S)—(N-(tert-Butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine (142 mg, 0.5 mmol) was dissolved in dichloromethane (50 mL). Dess-Martin periodinane (232 mg, 0.5 mmol) was added. After 1 hour at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. Purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether 60-80) to give a colourless oil identified as 1-[2-(S)—(N-(tert-butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine (40 mg, 0.14 mmol, 27%).

D. 1-[2-(S)—(N-(tert-butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino) butanoyl]piperidine

1-[2-(S)—(N-(tert-Butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine (40 mg, 14 mmol) was dissolved in methanol (20 mL). To this solution was added N-methyl-2-methylbenzylamine (19 mg, 0.14 mmol). After 2 hours sodium triacetoxyborohydride (64 mg, 0.3 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 1-[2-(S)—(N-(tert-butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino) butanoyl]piperidine (36 mg, 0.09 mmol, 64%).

E. 1-[2-(S)-Amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride

1-[2-(S)—(N-(tert-Butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino) butanoyl]piperidine (36 mg, 0.09 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 1-[2-(S)-amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride (43 mg, 0.09 mmol, 100%).

Example 1283

1-[N-(2″-(Cyclohexylmethylaminoethyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (2.5 g, 5.7 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (833 mg, 6.3 mmol), water-soluble carbodiimide (974 mg, 6.3 mmol), thiomorpholine (617 mg, 6.0 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine (2.7 g, 5.1 mmol, 90%).

B. 1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine

1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine (2.7 g, 5.1 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N-2′-(tert-butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine (1.44 g, 4.7 mmol, 92%).

C. 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine

1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine (100 mg, 0.3 mmol) was dissolved in methanol (25 mL). To this solution was added cyclohexanecarboxaldehyde (34 mg, 0.3 mmol). After 30 mins sodium triacetoxyborohydride (126 mg, 0.6 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine (33 mg, 0.08 mmol, 27%).

D. 1-[N-(2″-(Cyclohexylmethylaminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl-N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine (33 mg, 0.081 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-(cyclohexylmethylaminoethyl)glycinyl)]thiomorpholine dihydrochloride (31 mg, 0.08 mmol, 100%).

[M+H]⁺=300.3

Example 1284

1-[N-(2″-((Quinolin-2-ylmethyl)aminoethyl)glycinyl)]pyrrolidine dihydrochloride

A. 1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (2.5 g, 5.7 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.5 g, 11.1 mmol), water-soluble carbodiimide (1.3 g, 6.8 mmol), piperidine (484 mg, 5.69 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine (2.8 g, 5.5 mmol, 96%).

B. 1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine

1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine (2.8 g, 5.5 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N-2′-(tert-butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine (1.4 g, 4.9 mmol, 89%).

C. 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine

1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine was dissolved in methanol (25 mL). To this solution was added 2-quinolinecarboxaldehyde. After 30 mins sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine.

D. 1-[N-(2″-((Quinolin-2-ylmethyl)aminoethyl)glycinyl)]piperidine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl-N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-((quinolin-2-ylmethyl)aminoethyl)glycinyl)]piperidine dihydrochloride.

Example 1285

1-[N,N-(2″,2″-((Dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[2′-N-(tert-Butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine

(2S)-1-(N^α-(tert-Butyloxycarbonyl)-L-lysinyl)-pyrrolidine-2-carbonitrile (250 mg, 0.83 mmol) was dissolved in dichloroethane (25 mL). To this solution was added trans-cinnamaldehyde (108 mg, 0.83 mmol). After 30 mins sodium triacetoxyborohydride (350 mg, 1.6 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine. Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N-(2″-((cinnamyl)aminoethyl)-glycinyl]thiomorpholine (180 mg, 0.43 mmol, 52%).

B. 1-[N,N-(2″,2″-((Dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N,N-(2″,2″-((dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride.

Example 1286

1-[N-(2″-((Cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[N-(2″-((Cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl N-(2″-((cinnamyl)aminoethyl)-glycinyl]thiomorpholine (180 mg, 0.43 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-((cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride (168 mg, 0.43 mmol, 100%).

[M+H]⁺=320.3

Example 1287

3,3-Difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine dihydrochloride

A. 3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (1.0 g, 2.27 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (620 mg, 4.6 mmol), water-soluble carbodiimide (560 mg, 2.8 mmol), 3,3-difluoropyrrolidine hydrochloride (360 mg, 2.5 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine (934 g, 1.7 mmol, 77%).

B. 3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine (890 g, 1.68 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine (470 mg, 1.5 mmol, 91%).

C. 3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine (50 mg, 0.16 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (46 mg, 0.34 mmol), water-soluble carbodiimide (40 mg, 0.2 mmol), niflumic acid (49 mg, 0.17 mmol) and N-methylmorpholine (40 mg, 0.4 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine (63 mg, 0.11 mmol, 67%).

D. 3,3-Difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl) glycinyl)]pyrrolidine dihydrochloride

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine (55 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3,3-difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine dihydrochloride (52 mg, 0.10 mmol, 100%).

[M+H]⁺=472.3

Example 1288

3,3-Difluoro-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl) glycinyl)]thiomorpholine dihydrochloride

A. 4,6-Dichloro-2-(4′-fluoroanilino)-1,3,5-triazine

Cyanuric chloride (1.844 g, 10 mmol) was dissolved in acetonitrile (20 mL). The solution was cooled to −20° C. A solution of 4-fluoroaniline (1.1 g, 10 mmol) and triethylamine (1.0 g, 10 mmol) was slowly added. After 1 hour at −20° C. the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). The solution was washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was recrystallised from ethyl acetate/hexane to give an off white solid identified as 4,6-dichloro-2-(4′-fluoroanilino)-1,3,5-triazine 1.7 g, 6.0 mmol, 60%).

B. 1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine

1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]thiomorpholine (100 mg, 0.3 mmol) was dissolved in dichloromethane (30 mL). To this solution was added 4,6-dichloro-2-(4′-fluoroanilino)-1,3,5-triazine (90 mg, 0.3 mmol) and triethylamine (50 mg, 0.5 mmol). After 2 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). This solution was washed with water (2×30 mL) and brine (1×30 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine (20 mg, 0.032 mmol, 11%).

C. 1-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine (18.8 mg, 0.03 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride (18 mg, 0.03 mmol, 100%).

[M+H]⁺=526.4

TABLE 9
	Ex No	X	a	R
	128912901291129212931294	SCF2CHFSCH2O	1  2
	129512961297129812991300	SCF2CHFSCH2O	1  2
	131113121313131413151316	SCF2CHFSCH2O	1  2
	1317131813191320	SCF2CHFO	1  2
	132113221323132413251326	SCF2CHFSCH2O	1  2
	132713281329133013311332	SCF2CHFSCH2O	1  2
	133313341335133613371338	SCF2CHFSCH2O	1  2
	133913401341134213431344	SCF2CHFSCH2O	1  2
	134513461347134813491350	SCF2CHFSCH2O	1  2
	135113521353135413551356	SCF2CHFSCH2O	1  2
	135713581359136013611362	SCF2CHFSCH2O	1  2
	136313641365136613671368	SCF2CHFSCH2O	1  2
	136913701371137213731374	SCF2CHFSCH2O	1  2
	137513761377137813791380	SCF2CHFSCH2O	1  2
	138113821383138413851386	SCF2CHFSCH2O	1  2
	138713881389139013911392	SCF2CHFSCH2O	1  2
	139313941395139613971398	SCF2CHFSCH2O	1  2
	139914001401140214031404	SCF2CHFSCH2O	1  2
	140514061407140814091410	SCF2CHFSCH2O	1  2
	141114121413141414151416	SCF2CHFSCH2O	1  2
	141714181419142014211422	SCF2CHFSCH2O	1  2
	142314241425142614271428	SCF2CHFSCH2O	1  2
	142914301431143214331434	SCF2CHFSCH2O	1  2

TABLE 10
	Ex No	X	a	R
	1614161516161617	SCF2SCH2	1 2
	1618161916201621	SCF2SCH2	1 2
	1622162316241625	SCF2SCH2	1 2
	1626162716281629	SCF2SCH2	1 2
	1630163116321633	SCF2SCH2	1 2
	1634163516361637	SCF2SCH2	1 2
	1638163916401641	SCF2SCH2	1 2
	1642164316441645	SCF2SCH2	1 2
	1646164716481649	SCF2SCH2	1 2
	1650165116521653	SCF2SCH2	1 2
	1654165516561657	SCF2SCH2	1 2
	1658165916601661	SCF2SCH2	1 2
	1662166316641665	SCF2SCH2	1 2
	1666166716681669	SCF2SCH2	1 2
	1670167116721673	SCF2SCH2	1 2
	1674167516761677	SCF2SCH2	1 2
	1678167916801681	SCF2SCH2	1 2
	1682168316841685	SCF2SCH2	1 2
	1686168716881689	SCF2SCH2	1 2
	1690169116921693	SCF2SCH2	1 2
	1694169516961697	SCF2SCH2	1 2
	1698169917001701	SCF2SCH2	1 2
	1702170317041705	SCF2SCH2	1 2

Claims

1. A compound according to general formula 1, or a pharmaceutically acceptable salt thereof,

2. A compound according to general formula 8, or a pharmaceutically acceptable salt thereof,

3. A compound according to claim 2 wherein R1 is H.

4. A compound according to claim 2 wherein R1 is CN.

5. A compound according to any of claims 2 to 4 wherein X1 is CH2.

6. A compound according to any of claims 2 to 4 wherein X1 is S.

7. A compound according to any of claims 2 to 6 wherein b is 1.

8. A compound according to any of claims 2 to 6 wherein b is 2.

9. A compound according to any of claims 2 to 8 wherein a is 1.

10. A compound according to any of claims 2 to 8 wherein a is 2 and X2 is CH2.

11. A compound according to any of claims 2 to 10 wherein X3, X4 and X5 are all N.

12. A compound according to general formula 9, or a pharmaceutically acceptable salt thereof,

13. A compound according to claim 12 wherein R1 is H.

14. A compound according to claim 12 wherein R1 is CN.

15. A compound according to any of claims 12 to 14 wherein X1 is CH2.

16. A compound according to any of claims 12 to 14 wherein X1 is S.

17. A compound according to any of claims 12 to 16 wherein b is 1.

18. A compound according to any of claims 12 to 16 wherein b is 2.

19. A compound according to any of claims 12 to 18 wherein a is 1.

20. A compound according to any of claims 12 to 19 wherein X3, X4 and X5 are all N.

21. A compound according to general formula 10, or a pharmaceutically acceptable salt thereof,

22. A compound according to claim 21 wherein R1 is H.

23. A compound according to claim 21 wherein R1 is CN.

24. A compound according to any of claims 21 to 23 wherein X1 is CH2.

25. A compound according to any of claims 21 to 23 wherein X1 is S.

26. A compound according to any of claims 21 to 25 wherein b is 1.

27. A compound according to any of claims 21 to 25 wherein b is 2.

28. A compound according to any of claims 21 to 27 wherein a is 1.

29. A compound according to any of claims 21 to 27 wherein a is 2 and X2 is CH2.

30. A compound according to general formula 11, or a pharmaceutically acceptable salt thereof,

31. A compound according to claim 30 wherein R1 is H.

32. A compound according to claim 30 wherein R1 is CN.

33. A compound according to any of claims 30 to 32 wherein X1 is CH2.

34. A compound according to any of claims 30 to 32 wherein X1 is S.

35. A compound according to any of claims 30 to 34 wherein b is 1.

36. A compound according to any of claims 30 to 34 wherein b is 2.

37. A compound according to any of claims 30 to 36 wherein a is 1.

38. A compound according to general formula 12, or a pharmaceutically acceptable salt thereof,

39. A compound according to claim 38 wherein R1 is H.

40. A compound according to claim 38 wherein R1 is CN.

41. A compound according to any of claims 38 to 40 wherein X1 is CH2.

42. A compound according to any of claims 38 to 40 wherein X1 is S.

43. A compound according to any of claims 38 to 42 wherein b is 1.

44. A compound according to any of claims 38 to 42 wherein b is 2.

45. A compound according to any of claims 38 to 44 wherein a is 1.

46. A compound according to any of claims 38 to 44 wherein a is 2 and X2 is CH2.

47. A compound according to general formula 13, or a pharmaceutically acceptable salt thereof,

48. A compound according to claim 47 wherein R1 is H.

49. A compound according to claim 47 wherein R1 is CN.

50. A compound according to any of claims 47 to 49 wherein X1 is CH2.

51. A compound according to any of claims 47 to 49 wherein X1 is S.

52. A compound according to any of claims 47 to 51 wherein b is 1.

53. A compound according to any of claims 47 to 51 wherein b is 2.

54. A compound according to any of claims 47 to 53 wherein a is 1.

55. A pharmaceutical composition comprising a compound according to any of claims 1 to 54.

56. A use for a compound according to any of claims 1 to 54, which is as a component in the preparation of a pharmaceutical composition.

57. A method of treatment of disease in a human or animal subject, comprising a step of administering to the subject a therapeutically active amount of a compound according to any of claims 1 to 54.

58. A method of treatment according to claim 57 where the disease is caused by dysregulation of a post-proline cleaving proteases or their endogenous substrates.

59. A method of treatment according to claim 57 where the disease is ameliorated by inhibition of a post-proline cleaving proteases.

60. A method of treatment according to claim 57 where the disease is caused by dysregulation of a post-proline cleaving proteases or its endogenous substrates which is an intracellular protease.

61. A composition according to claim 1 or 38 with the proviso that when X1═S; b=1; R1═H; G2=H; G1 is —CH2—X2—(CH2)a-G3; a=1, X2═CH2; G3=NR15R16; and one of R15, R16═H, the other of R15, R16 is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.

62. A composition according to claim 1, 38, 47 or 60 with the proviso that when b=1, R1 is H and X1 is S; G1=H; G2 is —CH2—(CH2)a-G3; a=1; G3 is NR15R16 and one of R15 and R16 is H the other of R15, R16 is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.

63. A composition according to claim 1, 38, 47, 60 or 61 with the proviso that when b=1, R1 is CN and X1 is CH2; G1=H; G2 is —CH2—(CH2)a-G3; a=1; G3 is NR15R16 and one of R15 and R16 is H, the other of R15, R16 is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.

64. A composition according to claim 1, 38, 47, 60, 61 or 62 with the proviso that when G2=H; G1=-CH2-X2—(CH2)a-G3; X2 is CH2; a=1; G3=NR15R16 and R15═R16═H; b is not 2 when X1 is O or CH2, and b is not 1 when X1 is CH2.

65. A method of treatment according to claim 57 where the disease is caused by dysregulation of a non-membrane associated post-proline cleaving proteases such as QPP, DPP-8 and DPP-9 enzymes or their endogenous substrates.

66. A method of treatment according to claim 57 where the disease is ameliorated by inhibition of a non-membrane associated post-proline cleaving proteases such as QPP, DPP-8 and DPP-9 enzymes.

67. A method according to claim 64 where the compound is a selective inhibitor of non-membrane associated post-proline cleaving proteases.