US20080255126A1

US20080255126A1 - Inhibitors of post-proline cleaving proteases

Info

Publication number: US20080255126A1
Application number: US12/004,054
Authority: US
Inventors: David M. Evans; Doreen M. Ashworth
Original assignee: Ferring BV
Current assignee: Ferring BV
Priority date: 2001-10-23
Filing date: 2007-12-20
Publication date: 2008-10-16
Also published as: RU2345065C2; MXPA04003829A; US7335677B2; US20050043299A1; KR20040047943A; JP2005511541A; GB0125445D0; NO20042080L; AU2002336190B2; IL161191A0; NZ532048A; PL370220A1; EP1446116A1; RU2004110719A; ZA200402501B; HUP0401738A2; CA2464254A1; WO2003035057A1; CN1571667A

Abstract

Novel compounds that are inhibitors of one or most post-proline cleaving proteases, e.g. dipeptidyl peptidase IV, according to general formula (1). R¹is H or CN, X¹is O, S, CH₂, CHF, CF₂, CH(CH₃), C(CH₃)₂or CH(CN), and b is 1 or 2. G¹is H or a group according to the formula —CH₂—X²—(CH₂)_a-G³and G²is H or a group according to the formula —CH₂—(CH₂9_a-G³, provided that one of G¹and G²is H and the other is not H. X²is O, S, or CH₂, and a is 0, 1 or 2, provided that when a is 1 then X₂is CH₂. G³is 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, R¹is H or CN, X¹is O, S, CH₂, CHF, CF₂, CH(CH₃), C(CH₃)₂or CH(CN), and b is 1 or 2. G¹is H or a group according to the formula —CH₂—X²—(CH₂)_a-G³and G²is H or a group according to the formula —CH₂(CH₂)_a-G³, provided that one of G¹and G²is H and the other is not H. X²is O, S or CH₂, and a is 0, 1 or 2, provided that when a is 1 then X²is CH₂. G³is a group according to one of general formulae 2-4.
X³, X⁴and X⁵are either nitrogen N or CH, provided that at least two of X³, X⁴and X⁵are N. X⁶is either O or NH. R²is either H or alkyl. R³is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷and R⁸are 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)₂and CN. X⁷is CH₂, O, S or NH. R⁹is either H or alkyl. R¹⁰, R¹¹, R¹², R¹³and R¹⁴are 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)₂and CN. R¹⁵and R¹⁶are 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 R¹⁷is H, alkyl or aryl, or R¹⁵and R¹⁶together are a group according to one of general formulae 5-7.
R¹⁸is H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl or N(alkyl)₂, and R¹⁹is 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 R¹⁵and R¹⁶are both H then X¹may not be S or CH₂if 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 R¹is either a hydrogen atom H or a nitrile group CN. The group X¹is 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)₂and 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 G¹is either H or a group according to the formula —CH₂—X²—(CH₂)_a-G³and the group G²is either H or a group according to the formula —CH₂—(CH₂)_a-G³, provided that one of G¹and G²is H and the other is not H. The group X²is selected from O, S and CH₂. The integer a is 0, 1 or 2, provided that when a is 1 then X²is CH₂.
The group G³is 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, X⁴and X⁵are selected from nitrogen N and methine CH, provided that at least two of X³, X⁴and X⁵are nitrogen. Preferably X³, X⁴and X⁵are all nitrogen. The group X⁶is selected from O and NH. R²is selected from H and alkyl. R³is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷and R⁸are 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)₂and CN.
In general formula 3, the group X⁷is selected from CH₂, O, S and NH. R⁹is selected from H and alkyl. R¹⁰, R¹¹, R¹², R¹³and R¹⁴are 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)₂and CN.
In general formula 4, R¹⁵and R¹⁶are 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 R¹⁷is selected from H, alkyl and aryl, or R¹⁵and R¹⁶together are a group selected from general formula 5, general formula 6 and general formula 7.
In these general formulae, the group R¹⁸is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂, and the group R¹⁹is 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 R¹⁵and R¹⁶are both H then X¹may not be S or CH₂if 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, NO₂and 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 R¹is H.
In another preferred embodiment of the invention R¹is CN.
In another preferred embodiment of the invention X¹is CH₂.
In another preferred embodiment of the invention X¹is 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 X²is CH₂.
In another preferred embodiment of the invention a is 1.
In another preferred embodiment of the invention a is 1 and X²is CH₂.
In another preferred embodiment of the invention a is 2 and X²is 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 X²is CH₂Further preferred compositions having b=2 have X¹═CH₂or S, for example Example 38 of Table 2. Further preferred compositions having b=1 include G1 groups having a=0 or 1 and X²is CH₂. Further preferred compositions having b=1 have X¹═S or CH₂or 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,

wherein:

either G¹is —CH₂—X²—(CH₂)_a-G³and G²is H, or

G²is —CH₂—(CH₂)_a-G³and G¹is H;

G³is selected from a group according to general formula 2, a group according to general formula 3, and a group according to general formula 4;

a is 0, 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X²is selected from CH₂, O and S, provided that if a is 1 then X²is CH₂;

X³, X⁴and X⁵are selected from N and CH, provided that at least two of X³, X⁴and X⁵are N;

X⁶is selected from O and NH;

X⁷is selected from CH₂, O, S and NH;

R¹is selected from H and CN;

R²is selected from H and alkyl;

R³is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂;

R⁴, R⁵, R⁶, R⁷and R⁸are 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)₂and CN;

R⁹is selected from H and alkyl;

R¹⁰, R¹¹, R¹², R¹³and R¹⁴are 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)₂and CN;

R¹⁵and R¹⁶are independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and —CH₂-L-R⁷, or R¹⁵and R¹⁶together form a group according to general formula 5, general formula 6 or general formula 7;

R¹⁷is selected from H, alkyl and aryl;

R¹⁸is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂;

R¹⁹is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂;

L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄—;

d and e are selected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5; and

f is selected from 1, 2 and 3;

provided that when R¹⁵and R¹⁶are both H and b is 1 then X¹is not S or CH₂.

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

wherein:

a is 0, 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X²is selected from CH₂, O and S, provided that if a is 1 then X²is CH₂;

X⁶is selected from O and NH;

R¹is selected from H and CN;

R²is selected from H and alkyl;

R³is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂;

R⁴, R⁵, R⁶, R⁷and R⁸are 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)₂and CN.

3. A compound according to claim 2 wherein R¹is H.

4. A compound according to claim 2 wherein R¹is CN.

5. A compound according to any of claims 2 to 4 wherein X¹is CH₂.

6. A compound according to any of claims 2 to 4 wherein X¹is 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 X²is CH₂.

11. A compound according to any of claims 2 to 10 wherein X³, X⁴and X⁵are all N.

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

wherein:

a is 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X⁶is selected from O and NH;

R¹is selected from H and CN;

R²is selected from H and alkyl;

R³is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂;

13. A compound according to claim 12 wherein R¹is H.

14. A compound according to claim 12 wherein R¹is CN.

15. A compound according to any of claims 12 to 14 wherein X¹is CH₂.

16. A compound according to any of claims 12 to 14 wherein X¹is 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, X⁴and X⁵are all N.

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

wherein:

a is 0, 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X²is selected from CH₂, O and S, provided that if a is 1 then X²is CH₂;

X⁷is selected from O, S, CH₂and NH;

R¹is selected from H and CN;

R⁹is selected from H and alkyl;

R¹⁰, R¹¹, R¹², R¹³and R¹⁴are 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)₂and CN is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂.

22. A compound according to claim 21 wherein R¹is H.

23. A compound according to claim 21 wherein R¹is CN.

24. A compound according to any of claims 21 to 23 wherein X¹is CH₂.

25. A compound according to any of claims 21 to 23 wherein X¹is 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 X²is CH₂.

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

wherein:

a is 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X⁷is selected from O, S, CH₂and NH;

R¹is selected from H and CN;

R⁹is selected from H and alkyl;

31. A compound according to claim 30 wherein R¹is H.

32. A compound according to claim 30 wherein R¹is CN.

33. A compound according to any of claims 30 to 32 wherein X¹is CH₂.

34. A compound according to any of claims 30 to 32 wherein X¹is 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,

wherein:

a is 0, 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

X²is selected from CH₂, O and S, provided that if a is 1 then X²is CH₂;

R¹is selected from H and CN;

R¹⁵and R¹⁶are each independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and CH₂-L-R¹⁷;

or R¹⁵and R¹⁶together are a group according to general formula 5, a group according to general formula 6 or a group according to general formula 7;

R¹⁷is selected from H, alkyl and aryl;

L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄—;

d and e are selected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5; and

f is selected from 1, 2 and 3;

39. A compound according to claim 38 wherein R¹is H.

40. A compound according to claim 38 wherein R¹is CN.

41. A compound according to any of claims 38 to 40 wherein X¹is CH₂.

42. A compound according to any of claims 38 to 40 wherein X¹is 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 X²is CH₂.

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

wherein:

a is 1 or 2;

b is 1 or 2;

X¹is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O;

R¹is selected from H and CN;

R¹⁷is selected from H, alkyl and aryl;

L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄—;

d and e are selected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5; and

f is selected from 1, 2 and 3.

48. A compound according to claim 47 wherein R¹is H.

49. A compound according to claim 47 wherein R¹is CN.

50. A compound according to any of claims 47 to 49 wherein X¹is CH₂.

51. A compound according to any of claims 47 to 49 wherein X¹is 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; G¹is —CH₂—X²—(CH₂)_a-G³; a=1, X²═CH₂; G³=NR¹⁵R¹⁶; and one of R¹⁵, R¹⁶═H, the other of R¹⁵, R¹⁶is 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, R¹is H and X¹is S; G¹=H; G²is —CH₂—(CH₂)_a-G³; a=1; G³is NR¹⁵R¹⁶and one of R¹⁵and R¹⁶is H the other of R¹⁵, R¹⁶is 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, R¹is CN and X¹is CH₂; G¹=H; G²is —CH₂—(CH₂)_a-G³; a=1; G³is NR¹⁵R¹⁶and one of R¹⁵and R¹⁶is H, the other of R¹⁵, R¹⁶is 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; X²is CH₂; a=1; G3=NR¹⁵R¹⁶and R¹⁵═R¹⁶═H; b is not 2 when X¹is O or CH₂, and b is not 1 when X¹is 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.