MXPA99005961A - Peptidomimetic inhibitors of the human cytomegalovirus protease - Google Patents

Peptidomimetic inhibitors of the human cytomegalovirus protease

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Publication number
MXPA99005961A
MXPA99005961A MXPA/A/1999/005961A MX9905961A MXPA99005961A MX PA99005961 A MXPA99005961 A MX PA99005961A MX 9905961 A MX9905961 A MX 9905961A MX PA99005961 A MXPA99005961 A MX PA99005961A
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MX
Mexico
Prior art keywords
methyl
carboxamido
dimethyl
propyl
information
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Application number
MXPA/A/1999/005961A
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Spanish (es)
Inventor
D Bailey Murray
Fazal Gulrez
Lavallee Pierre
W Ogilvie William
Marcandrepoupart
Original Assignee
Bailey Murray
Boehringer Ingelheim (Canada) Ltd
Fazal Gulrez
Lavallee Pierre
Ogilvie William
Poupart Marcandre
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Application filed by Bailey Murray, Boehringer Ingelheim (Canada) Ltd, Fazal Gulrez, Lavallee Pierre, Ogilvie William, Poupart Marcandre filed Critical Bailey Murray
Publication of MXPA99005961A publication Critical patent/MXPA99005961A/en

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Abstract

Un compuesto de fórmula (I), en donde X es CF3, C2F5, 2-benzotiazolo, CF2CONHR6, CONHR6, en donde R6 es CH2C6H5, CH2(4-yodofenil),CH3, (CH2)2OCH2C6H5, CH2-2-benzimidazolo, CH2-(3,4-metilenodioxibenceno), CH(CH3)C6H5óCH(CH2CH3)C6H5;óX es 2-benzoxazolo-R7 en donde R7 es H, 4-CH3, 5-CH3, 6-CH3ó7-CH3;R1 es H CH3óCH2CH3;R2 es CH2CONH2, CH2CH2CONH2, CH2-tiazolo, CH2CON(CH3)2, CH2CO-(pirrolidino), CH2CH(CH3)2óCH2C6H5;R3 es Et, CH(CH3)2, C(CH3)3, adamantilo, CH2C(CH3)3óC(CH3)2CO2H;y R20 es COCH2C(CH3)3, COCH2CH2C6OH, COCH2CH(CH3)3, C02C(CH3)3, CONHC(CH3)3, COCH2N(CH3)2, CO(CH2)3CO2H, CO-(S)-CH(NH2)C(CH3)3, CO-(S)-CH{NHC(O)O-C(CH3)C(CH3)3, CO-(S)-CH (NHCO(CH2)5NHC(O)OC(CH3)3)C(CH3)3óCO-(S)-CH{NHCO(CH2)5NH2)C(CH3)3. Estos Compuestos sonútiles para el tratamiento de la infección de citomegalovirus humano.

Description

Inhibitors Pdptidomimetics of the Protease of the Human Cytomegalovirus Field of the Invention The present invention relates to compounds, compositions and methods for the treatment of human cytomegalovirus infection (HCMV). In particular, the present invention provides novel idomimetic peptide inhibitors of the HCMV protease.
BACKGROUND OF THE INVENTION Human Cytomegalovirus (HCMV) is a highly prevalent member of the herpes virus family that infects up to 80% of the general population. This virus is responsible for opportunistic infections in immunocompromised individuals that include recipients of organ transplants, patients with cancer and patients suffering from AIDS. Clinical manifestations include disseminated disease, pneumoñitis, retinitis and gastro-intestinal infections such as esophagitis and colitis. Of particular importance are HCMV infections in neonates. This Ref.: 30570 Disease is the most common congenital viral infection acquired in the world. It is estimated that 1% of newborn infants are infected and up to 10% of these are symptomatic and may experience several complications. The mortality in this last group approaches 30%.
All members of the herpes virus family express a late protein in the life cycle of the virus that seems to function as a self-assembly ladder of the viral capsule. The assembly protein is present in the immature B capsules and must be processed to remove a short segment of the C term to allow the entry of the viral DNA and produce an infectious virus particle. Recently it has been shown that this processing is mediated by a protease that is encoded by the virus. The protease itself is expressed as a precursor protein that is autocatalytically cut at least twice (Scheme 1). The cut is presented near the C terminus of the UL80 gene product (site M) to remove a small fragment, and also at a position located near the center of the precursor (site R) to cut the catalytic domain (N0). The Ns and the full-length protease (product of the UL80 gene) are catalytically active.
SCHEME 1 Catalytic domain (N0) The HCMV Protease No shows the homology of the significant sequence with other proteases of the herpes virus. The affinity tagging and directed site mutagenesis experiments indicate that this enzyme is a serprotease. Recent crystallographic results have shown that the HCMV protease represents a new structure of serproteases and in fact possesses a unique catalytic triad.
While it has not been shown that the HMCV protease is absolutely required for viral replication, it has been shown that HSV-1 mutants lacking the analogous enzyme or expressing defective variations are unable to grow. The high degree of homology between the HSV and HCMV proteases maintains the idea that HCMV-specific protease inhibitors would show antiviral activity and thus have therapeutic value.
EP 0,410,411 A2 exhibits new peptidase inhibitors. These peptide analogs contain a pentafluoroethyl ketone in Pl ', however none of the exposed peptides contain the P2 amino acid derivatives set out in the present invention.
A.H. Abuelyaman et al. (Bioco ugate Chemistry, vol.5, no.5, October 1994, pp.400-405) discloses fluorescent peptide phosphonates. However, none of the exposed peptides corresponds or leads to the peptide derivatives of the invention.
Derstet al. (J. Am. Chem. Soc, vol 118, No. 35, 4 September 1996, pp. 8485-8486 discloses a number of peptidyl trifluoromethyl ketones. None of these peptides corresponds or leads to the peptide derivatives of the invention.
Brief Description of the Invention In accordance with the present invention, a compound of the formula I is provided: (I) where z is C or P; when z is C, then X is CF3; C2F5; benzothiazole; oxazolo [4, 5b] pyridine; or benzoxazole-R7 wherein R is H or methyl; OX is CF2CONH-R6, C (0) NH-R6, wherein R6 is C0-? Alkyl or optionally substituted with phenyl or cyclohexyl, the phenyl ring or cyclohexyl is optionally substituted with Me, halogen, -CF3, -CH ( Me) -C (0) - OBn; -C (0) NH2; or -C (O) -morpholino; the phenyl or cyclohexyl ring is optionally fused with phenyl ring; (CH2) 1-3-O- (CH2) 1-3-phenyl The phenyl is optionally substituted with halogen; (CH2) 1-3-2-benzimidazole; (CH2)? _3- (3,4-methylenedioxybenzene); or (CH2) 1-3 - 0 - C (0) - 0CH2 CH = CH2 i or, when z is P, then X is - (0Ph) 2; Ri is H, Me, or Et; R2 is CH2-S02NH2; alkyl -C? -6; - (C? _6 alkyl) aryl; - (C? -6 alkyl) thiazolo; -CH2C (O) - (C? _e alkyl); CH2C (0) -pyrrolidino; -CH2C (O) -morpholino; - (C? _6 alkyl) amino; - (C? _6 alkyl) amido optionally mono- or disubstituted with C? Alkyl, the alkyl optionally substituted with pyridino; is NH, CH2 or CH (CH3); R3 is alkyl -C? _? 2; - (C? _6 alkyl) C (0) OH; or adamant ilo; n is 0 or 1, R *, when n is 1, is alkyl -C? -6 or - (C? _6 alkyl) -aryl wherein the aryl is optionally substituted with OH; it's 0 or 1 R5, when m is 1, is H or -CH2OH; And it is H; (CH2) 2-t-Bu; or an acyl of the formula: -C (0) - (CH2)? _6-C (0) 0H; -C (0) - (CH2)? _6 -Ph where Ph is optionally substituted with OH; -C (0) -CH2N (CH3) 2; -C (0) -R9; -C (0) 0-R9; or -C (0) NH-R9 wherein R9 is C6-C6 alkyl; or -C (O) - (CH2)? -6-NH2 wherein the amino group is optionally protected with an amino protecting group.
Included within the scope of this invention is a pharmaceutical composition comprising a quantity of virally effective anti-cytomegalovirus of a compound of the formula I or a pharmaceutically acceptable salt thereof, mixed with a pharmaceutically acceptable carrier medium or auxiliary agent.
An important aspect of the invention involves a method for treating viral infection by cytomegalovirus in a mammal by administering to the mammal a virally effective anti-CMV amount of the compound of formula I or a therapeutically acceptable salt thereof, or a composition as described above. .
Another important aspect involves a method for inhibiting the replication of cytomegalovirus virus by exposing the virus to an amount of CMV protease inhibition of the compound of the formula I or a therapeutically acceptable salt thereof, or a composition as described above.
Preferred compounds of the invention include the compounds of the formula I: wherein the substituents are subsequently defined Preferably, z is C. or P More preferably, z is C.
Preferably, X is CF3; C2 F5; 2-ben zot-iazole; 2-oxazolo [4, 5b] pyridine; 2-benzoxazole-R, wherein R is H, 4-Me, 5-Me, 6-Me, or 7-Me; CF2C0NHR6 or C (0) NHR6 wherein Re is C? _ Alkyl, optionally substituted with cyclohexyl, naphthyl, or phenyl optionally substituted with Me, iodo, CF3, -CH (Me) -C (O) -OBn; -C (0) NH2, or C (O) -morpholino; (CH2) 2-0-CH2-phenyl; CH2-2-benzimidazole; or CH2- (3,4-methylenedioxy benzin); or when z is P, then X is (0Ph) 2 « More preferably, X is CF3; C2F5; benzothiazole; benzoxazole-R7, wherein R7 is H, 4-Me, 5-Me, 6-Me, or 7-Me; -CF2C0NH-CH2-phenyl; -C (0) NHR6 wherein R6 is -CH (Me) (CH2) 4CH3; cyclohexyl; naphthyl; -CH2-phenyl; -CH (CH 3) -phenyl; or -CH (CH2CH3) -phenyl; -CH2-4-iodophenyl; -phenyl-CH3; -phenyl-CF3; -f-C-enyl (0) NH2; -phenyl-C (O) -morpholino; -f-enyl-CH (Me) -C (0) -OBn; - (CH2) 2-0- CH2-phenyl; -CH2-2-benzimidazole; -CH2- (3,4-methylenedioxybenzene); or - (CH2) 2-0-C (0) - OCH2CH = CH2; or when z is P, then X is (OPh) 2.
More preferably, X is C2F5; -C (0) NHR6 wherein R6 is -CH2-phenyl; -CH2-4-iodophenyl; CH (CH 3) -phenyl; or -CH (CH2CH3) -phenyl; CH (Me) -naphthyl; -CH2CH (Me) -phenyl; - (CH2) 2-0- CH2-phenyl; -CH2-2-benzimidazole; or -CH2- (3,4-methylenedioxybenzene); Preferably Ri is H, methyl or ethyl. More preferably, Ri is H or methyl. More preferably, Ri is H or methyl; Preferably, 2-CH2- (4-thiazoloi) - (CH2)? - 4-NH2; -CH2-C (O) -ter-butyl; -CH2-C (O) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); -CH2S02NH2; _ (CH2)? -2-ami or, the amide nitrogen optionally mono- or di-substituted with a substituent independently selected from: CH3; t-Bu; phenyl; or -CH2CH2- (2-pyridino).
More preferably, R2 is -CH2-C (O) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); -CH2S02NH2; - (CH2) C (0) NH2; - (CH2) 2C (0) N (CH3) 2; -CH2-C (O) -NH-t-Bu; or - (CH2) 2-C (O) -N (CH3) CH2CH2 (2-pyridino). More preferably, R2 is -CH2-C (0) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); - (CH2) 2C (0) N (CH3) 2; or - (CH2) 2-C (0) -N (CH3) CH2CH2 (2-pyridino).
Preferably, it is NH or CH2. More preferably, W is NH.
Preferably, R3 is ethyl; isopropyl; t-Bu; CH2-t-Bu; or adamantyl. More preferably, R3 is ethyl; isopropyl; or t-Bu. More preferably, R3 is isopropyl; or t-Bu.
Preferably, n is 0 or 1 More preferably, n is 0. Alternatively, more preferably, n is 1 Preferably, R, when n is 1, is isopropyl; t-Bu; or 4-hydroxybenzyl. More preferably, R, when n is 1, is isopropyl; or t-Bu. More preferably, R4, when n is 1, is t-Bu; Preferably, it is 0 or 1. More preferably, m is 0.
Preferably, R5, when m is 1, is H.
Preferably, Y is H; • CH2-CH2-t-Bu; an acyl of the formula: -C (0) CH3; -C (0) CH2-CH (CH3) 2; -C (0) CH2-t-Bu (DA-Tbg); -C (0) (CH2) 2-4-hydroxyphenyl; -C (O) - (CH2) 3 -COOH; -C (0) 0-t-Bu (Boc); -C (0) NH-t-Bu; -C (0) CH2-N (CH3) 2; or -C (O) (CH2)? 6NH2, the ammo group is optionally protected with an amino protecting group More preferably, Y is H; or an acyl of the formula: -C (0) CH3; -C (0) CH2-CH (CH3) 2; -C (O) CH2-t-Bu (DA-Tbg); -C (O) (CH2) 2-4-hydroxyphenyl; -C (0) - (CH2) 3 -COOH; -C (0) 0-t-Bu (Boc); -C (O) (CH2) 5 NH2; O -C (O) (CH2) 5NH-Boc.
More preferably, it is an acyl of the formula: -C (O) CH2-t-Bu (DA-Tbg); -C (O) O-t-Bu (Boc); -C (O) (CH2) 5 NH2; or -C (O) (CH2) 5NH-B? c.
A preferred compound of the invention is selected from the group consisting of: NI - (3, 3, 3-trifluoro-1-met il-2-oxopropyl) - (2 S) -2- ((1 S) -2-methyl-l- [((SS) -2-methi 1 -1- [(Methylcarboxamido) methyl] carboxamidopropyl) carboxamido] propylcarboxamido) butanediamide (37) NI- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -6-amino-2- ( (15) -1 - [((15) -1 - [(1S) -2-hydroxy-1- (methylcarboxamido) ethyl] carboxamido-2- (4-hydroxyphenyl) ethyl) carboxy-ido] -2-methylpropyl-carboxamide ) hexanamide (38); NI- (3, 3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2- [((1S) -2-methyl-1 - [(1S) -2-methyl-1- ( methylcarboxamido; propyl] carboxamidopropyl) carboxamido] butanediamide (39); NI- (3, 3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2-. { (SS) -2-methyl-l- [(methylcarboxamido) -propyl] carboxamido} butanediamide (40); NI- (3, 3, 3-trifluoro- (SS) -methyl-2-oxopropyl) - (25) -2-. { (15) -2-methyl-l- [(methylcarboxamido) propyl] carboxamido} butanediamide (43); NI- (1-ethyl-3, 3, 3-trifluoro-2-oxopropyl) - (25) -2- [((15) -2-methyl-l- [(15) -2-methyl-1- ( methylcarboxamido) propyl] carboxamidopropyl) carboxamido] utanediamide (44); Nl- (l- (3,3,3, -trifluoro-1-propyl-2-oxopropyl) - (2S) -2- [((15) -2-methyl-l- [(15) -2-methyl) -l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (45); NI- (3, 3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2- [((15) -2-methyl-1 - [(15) -2-methyl-1- ( methylcarboxamido) propyl] carboxamidopropyl) carboxamido] pentanediamide (46); Acid (35) -3- [((15) -2-methyl-l- [(15) -2-methyl-l- (methylcarboxamyl) propyl] carboxamido-propyl) carboxamido] -3- [(3.3 , 3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] propanoic (47); Nl - [(15) - 1 - ((15) -2-hydroxy-1- [(3, 3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] ethyl-car amoyl) -2-methylpropyl] - (25) -3-methyl-2- (methylcarboxamido) butanamide (48); NI - (3, 3, 3-trifluoro-1-methyl-2-oxopro-yl) - (25) -6-amino-2- [. { (1S) -2-methyl-l- [(15) -2-methyl-1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] hexanamide (49); NI - [(15) -2-methyl-l- ((15) -2- (1,3-thiazol-4-yl) -l- [(3,3,3-t-fluoro-1-methyl) 1-2-oxopropyl) -carbamoyl] ethylcarbamoyl) propyl] - (25) -3-met il-2- (methylcarboxamido) utanamide (50); N4, N4 -dimet i l -Nl -. { 3, 3, 3-trifluoro-l-methyl-2-oxopropyl) - (25) -2 - [((lS) -2-methyl-l- [(15) -2-methi 1-1 - (methylcarboxamido propyl] carboxamidopropyl) carboxamido] butanediamide (51); NI - (3, 3, 3-trifluoro-1-met i 1-2 -oxopropyl) - (25) -4-methyl-2- [((SS) -2-methyl-1 - [(15) -2 -methyl 1-1 - (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] pentanamide (52); NI - [(15) -2-methyl-l- ((lS) -2-phenyl-1 - [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) propyl] - (25) -3-met i 1-2- (methylcarboxamido) butanamide (53); NI - [(1 S) -2-methyl-l- ((15) -2-methyl-l- [(3,3,3-trifluoro-1-met i 1-2 -oxopropyl) carbamoyl] -propyl carbamoyl ) propyl] - (25) -3-met i 1-2- (methylcarboxamido) butanamide (54); NI - [(15) -2-methyl-l- ((15) -l- [(3,3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] ethyl -carbamoyl) propyl] - (25) -3-methyl-2- (methylcarboxamido) butanamide (55); NI- [(15) -2-methyl-l- ((1.) -1- [(3,3, 3-trifluoro-1-met il-2-oxopropyl) carbamoyl] et i 1 -carbamoyl) propyl] - (25) -3-methyl-2- (methylcarboxamido) butanamide (56); N 4, N 4 -dimethyl-N 1 - (3, 3, 3-trifluoro-1-methy1-2-oxopropyl) - (25) -2 - [((15) -l - [(lS) -2-methi 1- 1- (Methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (57); N4, N4-imethyl-Nl- (3,3, 3-trifluoro-1-met i 1-2-oxopropyl) - (25) -2 - [((lS) -2, 2 -dimethyl-1- [( 15) -2-methyl-1- (methylcarboxamido) propyl] carboxamido-propyl) carboxamido] butanediamide (58); N 4, N 4 -dimethyl-N 1 - (3,3,3-t-fluoro-1-met i 1-2-oxopropyl) - (25) -2- [((15) -3, 3 -dimet i 1- 1- [(15) -2-methyl-1- (methylcarboxamido) propyl] carboxamidobutyl) carboxamido] butanediamide (59); N4, N4 -dimet il-Nl- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2 - [((5) -l- (1 -adama t il) -1 - [(15) -2-methyl-1- (methylcarboxamido) propyl] carboxamido methyl) carboxamido] butanediamide. { 60); Acid (35) -3- ((15) -2- (dimet i 1carbamoyl) -1- [(3, 3, 3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) -2, 2- dimethyl-3- [(15) -2-methyl-1-1- (methylcarboxamido) propyl] carboxamidopropanoic acid (61); N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-1-meth i 1-2-oxopropyl) - (25) -2 - [(15) -2, 2-dimethyl-1- (ethylcarboxamido) propyl] carboxamidobutanediamide (62); N 4, N 4 -diraethyl-N 1 - (3,3, 3-trifluoro-1-methyl-2-oxo-propyl) - (25) -2 - ((15) -l- [(4-hydroxyphenyl-ethyl) carboxamide] -2, 2-dimethylpropylcarboxamido) butanediamide (63); N4, N4 -dimethyl -NI- (3,3, 3-tri-fluoro-1-met i 1-2 -oxopropyl) - (25) -2- [(15) -l- (isobutylcarboxamido) -2, 2 -dimethylpropyl] carboxamidobutanediamide (64); N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-1-met il-2-oxopropyl) - (25) -2 - [(15) -2,2-dimethyl-l- (eopent i 1 carboxamido) propyl] carboxamidobutanediamide (65); N4, N4 -dimeti 1-N1- (3,3, 3-trif1uoro-1-methyl-2-oxopropyl) - (25) -2- ((15) -1- [(3, 3 -dimet i1-but il) amino] -2, 2-dimethyl-propylcarboxamido] butanediamide (66); 4N, 4N-Dimethyl-1N- (3,3,3-trifluoro-1-methyl-2-oxo-propyl) -2- [1- (ex-butoxycarbonyl-amino) -2,2-dimethyl- (SS) - propyl carboxamido] - (2S) -butandiamide (67); N4, N4-Dimethyl-Nl- (3,3, 3-trifluoro-1-met il-2-oxopropyl-2- [l- (tert-butyl lami ocarbonyl-amino) -2,2-dimethyl- (SS) -pro-ilcarboxamido] - (2S) -butandiamide (68); N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-1-methyl-yl-2-oxopropyl) - (25) -2- [((15) -1- [(dimethylamino) methyl] carboxamido -2, 2 -dimeti lpropi 1) carboxamido] butanediamide (69); 4-t (15) -l- ((15) -2- (dimet i 1carbamoyl) -1- [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] ethylcarbamoyl) -2 acid 2-dimet i lpropi 1] carbamoylbutanoic (70); N4, N4-dimeth yl-NL- (3,3,4,4, 4 -pentafluoro- 1-meth 1-2-i oxobutyl) - (25) -2- [(IS) -2, 2-dimethyl- 1- (neopentyl carboxamido) propyl] carboxamidobutanediamide (74); NI- [3- (benzylcarbamoyl) -3, 3 -difluoro- 1 -methyl-2-oxopropyl] -N4, N4 -dimethyl- (25) -2- [(15) -2, 2 -dimet i 1-1 - (neopentylcarboxamido) propyl] carboxamidobutanediamide (75); Benzyl acid amide 3-. { 2- [3- (3, 3-Dimethyl-butyrylamino) -3,3-dimethyl-butyrylamino] -3-dimethylcarbamoyl-propionylamino} -2-oxo-butyric (76); Nl- [2- (1, 3-benzoxazol-2-yl) -l-methyl-2-oxoethyl] -N 4, N 4 -dimethyl- (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (77); Diphenyl N4, N4-dimethyl-Nl- (1-aminoethylphosphinate) - (25) -2-. { [(15) -2, 2-dimeti 1-1- (neopent i 1carboxamido) propyl] carboxamido} butan diamide (79); NI- [2- (1, 3-benzothiazol-2-yl) -l-methyl-2-oxoetyl] -N 4, N 4 -dimethyl- (2S) -2-. { [(15) -2,2-di-met il-1 - (neopentyl carboxamido) propyl] carboxamido} utan diamide (80); N 4, N 4 -dimethyl-N 1 - (1-met i 1-2- [1,3] oxazolo [4,5-J] pyridin-2-yl-2-oxoethyl) - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (81); N 4, N 4 -dimet il-N 1 - [1-methy1-2- (6-methyl-1,3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimeti 1-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (82); N 4, N 4 -dimet il-N 1 - [1-methy1-2- (5-methyl-1,3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2,2-dimethyl-l- (neopentylcarboxamyl) propyl] carboxamido} butanediamide (83); N 4, N 4 -dimet il-N 1 - [1-methy1-2- (4-methyl-1,3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimeti 1-1- (eopentylcarboxamido) propyl] carboxamido} butanediamide (84); N4, N4-dimethyl-Nl- [1-methyl-2- (7-methyl-1-, 3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (85); N 4, N 4 -dimet il-N 1 - [1-met i 1-2- (met i 1carbamoyl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimethy1-1- (neopentyl carboxamido) propyl] carboxamido} butanediamide (86); NI- (2- [2- (benzyloxy) ethyl] carbamoi 1-1-methyl-2-oxoethyl) -N 4, N 4 -dimethyl- (25) -2-. { [(15) -2, 2-dimethyl-l- (neopentylcarboxamido) propyl] carboxamido} butanediamide (88); Nl-2- [(1,3-benzodioxol-5-ylmethyl) carbamoyl] -1-methyl 1-2-oxoethyl-N4, N4-dimethyl- (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (89); Nl-2- [(IIH-be zo [] imide zol-2-methylmethyl) carbamoyl] -1-methyl-2-oxoethyl-N4, N4-dimethyl- (25) -2-. { [. { 15) -2,2-dimeti 1-1- (neopentylcarboxamido) propyl] carboxamido Jbutanediamide (90); N 4, N 4 -dimet il-N 1 - (1-met il-2-oxo-2- [(15) -1-phenylethyl] carbamoylethyl) - (25) -2-. { [(15) -2, 2 -dimet i 1-1- (neopentylcarboxa) propyl] carboxamido} butanediamide (91); N 4, N 4 -dimet il-N 1 - (1-methyl-2-oxo-2- [[IR) -1-phenilethyl] carbamoylethyl) - (25) -2-. { [(15) -2,2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (92); N 4, N 4 -dimet il-N 1 - (1-methyl-2-oxo-2- [(li?) -1-phenylpropyl] carbamoyl-ethyl) - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (93); 95; 96; Ac-Ser-Tyr-Val-Lys-Ala (d, 1) -C (0) -NH-CH2-Ph 218; 301; 304; O O .OBn DA-Tbg-Tbg- C7 305; 307; 308; 312; 402; 405, 408; 409; 411; 412; 413; Y A i? O? Ar 414 A further aspect of the present invention is a solid phase process for the synthesis of peptidyl activated ketones comprising the steps of: a) coupling an icarbazone acid of formula 113 to a resin by in-situ activation; 113 II 103 wherein R is a side chain of a natural or non-natural amino acid; and X 'is CF3, CF2CONH-R30, C (0) NH-R30, or C (0) OR 30, wherein R30 is an alkyl C3_? 2 cyclic or C? -10 acyclic alkyl or alkenyl C3_? 2 cyclic or alkenyl C2_? 2 acyclic, the alkyl or alkenyl is optionally substituted with NH2, OH, SH, halo, or carboxyl; the alkyl or alkenyl optionally contains at least one heteroatom independently selected from the group consisting of: O, S, and N; or R3o is an aryl or C7 or C7_6 aralkyl optionally substituted with C6_6 alkyl, NH2, OH, SH, halo, carboxyl or carboxyalkyl (lower); the aryl or aralkyl optionally contains at least one heteroatom independently selected from the group consisting of: O, S, and N; A is a divalent spacer group comprising a non-reactive divalent hydrocarbyl group having from 2 to 15 carbon atoms; Y Pg is an amino protecting group b) deprotecting the amino protecting group to give the desired resin of the formula 103; c) coupling the resin with one or more amino acids in a sequential manner by standard chemistry; and d) cutting the peptide from the resin to obtain a peptidyl-activated ketone of the formula II.
Preferably, the cutting step as described herein is carried out in THF, ac HCl, and AcOH at a temperature of about 60 ° C for about 4 hours; and the resin is filtered at least once.
Preferably, the resin is selected from the group consisting of: polystyrene or pegylated polystyrene functionalized with benzydrylamine (BHA); 4-methyl benzidrilamine (MBHA); and aminomethyl (AM).
Preferably, the activation in si tu is carried out with the addition of a coupling agent selected from the group consisting of: 2- (IH-benzotriazol-1-yl) -1, 1, 3, 3-tetramet illuronium tetrafluoroborate (TBTU); 2- (Li? -benzotriazol-1-yl) -1, 1,3,3-tetramethyluronium hexafluorophosphate (HBTU); diisopropyl carbodiimide (DIC), and dicyclohexyl carbodiimide (DCC).
Preferably, the amino protecting group is selected from the group consisting of: t-butyloxycarbonyl (Boc); 9-fluorenylmet-iloxy carbonyl (Fmoc); and allyloxy carbonyl (Alloc).
Preference is given to C (0) NH2CH2 - f in i l o C (0) 0CH2 CH = CH2.
Preferably, R is selected from the group consisting of: CH3; CH2CH3; CH2CH2CH3; (CH2) 4 NH2; CH (CH3) 2; CH2-phenyl; (CH2) 3-NH-CH = N (NH2).
Preferably, A is cyclohexyl, phenyl or benzyl. Alternatively, a further aspect of the present invention is a resin of the formula 103 as defined above.
Yet, a further aspect of the present invention is the use of a resin of formula 103 for solid phase synthesis of peptidyl-activated ketones.
Detailed description of the invention As used here, the following definitions apply unless otherwise noted: With reference to the examples where (R) or (5) are used to designate the configuration of a radical P &R4 of the compound of formula I, the designation is made in the context of the compound and not in the context of the radical alone.
Natural amino acids, with the exception of glycine, contain a chiral carbon atom. Unless specifically indicated otherwise, compounds containing natural amino acids with the L configuration are preferred. However, applicants contemplate that when specified, some amino acids of formula I may be either of the D or L configuration or they can be mixtures of the D and L isomers, which include the epimeric mixtures 1: 1.
Non-natural amino acids include, but are not limited to, α-aminoadipic acid, α-α-diamino butyric acid, ornithine, pipecolic acid, sarcosine, thyroxine, hydroxylysine and hydroxyproline.
The abbreviations for some a-amino acids are set forth in Table A.
Table A As used herein, the term "tert-butylglycine" refers to a compound of the formula: The term "side chain" with reference to an amino acid or amino acid derivative means a residue bound to the α-amino acid α-amino acid atom. For example, the side chain of the R group for glycine is hydrogen, for alanine it is methyl, for asparagine it is CH2-C (0) NH2, for glutamine it is CH2CH2C (0) NH2, and ter-butylglycine is ter- butyl. For the specific R groups or side chains of the -amino acids, reference is made to the text of A.L. Lehninger in Biochemistry (see chapter 4).
The term "halo" as used herein means a halogen radical selected from bromine, chlorine, fluorine or iodine.
The term "C alquilo _ ?alkyl" or "(lower) alkyl" as used herein, either alone or in combination with another radical, means cyclic or acyclic alkyl radicals (straight or branched chain) containing up to ten atoms carbon and includes, for example, methyl, ethyl, propyl, butyl, hexyl, 1-methylethyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl. Obviously, as will be readily recognized by a person skilled in the art when a cycloalkyl is contemplated, unless otherwise indicated, the alkyl radical will contain at least 3 carbon atoms.
The term "C2-alkenyl" or "as used herein, either alone or in combination with another radical, means an alkyl radical as defined above of 2 to 10 carbon atoms, and further contains at least one double bond. For example, alkenyl includes allyl.
The term "Ce or Cly aryl" as used herein, either alone or in combination with another radical, means either an aromatic monocyclic system containing 6 carbon atoms or an aromatic bicyclic system containing 10 carbon atoms. For example, aryl includes phenyl or naphthalene.
The term "C7-a6-aralkyl" as used herein, either alone or in combination with another radical, means an aryl as defined above linked through an alkyl group, wherein alkyl is as defined above containing 1 to 6 carbon atoms. Aralkyl includes, for example, benzyl, and butylphenyl.
The term "divalent spacer group" as used herein means a non-reactive divalent hydrocarbyl group of 2 to 15 carbon atoms and includes, but is not limited to, cyclohexane, phenyl, and benzyl.
The term "heterocycle" as used herein, either alone or in combination with another radical, means a monovalent radical derived by the removal of a hydrogen from a saturated, unsaturated, five, six or seven membered heterocycle containing from one to four heteroatoms selected from nitrogen, oxygen and sulfur. Examples of the appropriate heterocycles include pyrrolidine, pyridine, thiazole, thiazolidine, benzothiazole, benzoxazole, benzimidazole, and 3,4-methylenedioxybenzene.
ANTIVIRAL ACTIVITY The antiviral activity of the aforementioned idomimetic peptide inhibitors of the HCMV protease (HCMV protease inhibitors) can be demonstrated by biochemical, chemical, microbiological and biological methods. For example, a test based on the evaluation of the ability of the test compound to inhibit the protease of HCMV, an enzyme vital for viral replication.
When the HCMV protease inhibitor is used as an antiviral agent, it is administered orally, or systemically to humans in a vehicle comprising one or more pharmaceutically acceptable carriers, the proportion of which is determined by the solubility and chemical nature of the drug. compound, the chosen route of administration and the standard biological practice. For oral administration, the compound or a pharmaceutically acceptable salt thereof can be formulated into unit dosage forms such as capsules or tablets each containing a predetermined amount of the active ingredient, in the range of about 50 to 500 mg, in a pharmaceutically vehicle. acceptable.
For parenteral administration, the HCMV protease inhibitor is administered either intravenously, subcutaneously or intramuscularly, in compositions with pharmaceutically acceptable carriers or carriers. For administration by injection, it is preferred to use the compounds in solution in a sterile aqueous vehicle which could also contain other solutes such as buffers or preservatives in addition to sufficient quantities of the pharmaceutically acceptable salts of glucose to make the solution isotonic.
Suitable carriers or carriers for the formulations noted above are described in standard pharmaceutical texts, e.g. ex. in "Remington's The Science and Practice of Pharmacy", 19th ed., Mack Publishing Company, Easton, Penn., 1995, or in "Pharmaceutical Dosage Forms and Drugs Delivery Systems," 6th ed., H.C. Ansel et al., Eds., Williams & Wilkins, Baltimore, Maryland, 1995.
The dosage of the protease inhibitor HCMV 'will vary with the form of administration and the particular active agent chosen. In addition, it will vary with the particular guest under treatment. In general, the treatment starts with small increments until the optimum effect is reached under the circumstance. In general, the inhibitor compound is most desirably administered at a concentration level that will generally provide antivirally effective results without causing any harmful or deleterious side effects.
For oral administration, the HCMV protease inhibitor is administered in the range of 20 to 200 mg per kilogram of body weight per day, with a preferred range of 25 to 100 mg per kilogram.
For ocular administration, the HCMV protease inhibitor is administered either topically or intraocularly (injection or implant) in an appropriate preparation. For example, an implant containing the compound in an appropriate formulation can be surgically placed in the posterior segment of the eye by means of a small incision.
With reference to systemic administration, the HCMV protease inhibitor is administered at a dosage of 10 mg to 150 mg per kilogram of body weight per day, although the aforementioned variations will be presented. However, a dosage level that is in the range of about 10 mg to 100 mg per kilogram of body weight per day is more desirably employed to achieve effective results.
CHEMISTRY The synthesis of several inhibitors and the required intermediates are described in Schemes 2 to 7.
Inhibitors containing a trifluoromethyl ketone function were obtained in one of three ways: solution chemistry or solid phase synthesis: schemes 2 or 3. Inhibitors containing a -ketoamide were obtained in one of two ways: solid phase: scheme 3 or solution chemistry: scheme 4. Inhibitors containing other activated ketones were obtained by solution chemistry: scheme 5.
Solid phase synthesis.
Scheme 2: Inhibitors that incorporate an asparagine residue in P2 (Scheme 2) could be prepared by solid phase synthesis using the side chain of asparagine as a point of attachment to the resin (Abraham, NA, Fazal, G., Ferland, J.-M .; Rakhit, S., Gauthier, J., A new solid phase strategy for the synthesis of mammalian glucagon, Te trahedron Le tt., 1991, 32, 577-580).
Scheme 2 __ 1 2 R = H, Me, Et Thus a Henry reaction between hemiacetal 1 and nitroethane gave the nitro alcohol 2 which was immediately reduced and protected to produce alcohol 3. After removal of the Boc group, coupling with an appropriately protected aspartic acid derivative gave 4. This compound was then deprotected by hydrolysis and incorporated into a polymer support to provide the derivatized amide resin 5. The required amino acids were then introduced by standard methods. The hydrolysis of the resin and the oxidation of the resulting alcohol using the method of Moffatt [(a) Pfitzner, K.E .; Moffatt, J.G. Sulfoxide-carbodiimide reactions. I. A facile oxidation of alcohols. J. Am. Ch em. Soc. 1965, 8 7, 5661-5670. (b) Pfitzner, K.E .; Moffatt, J.G. Sulfoxide-carbodiimide reactions. II. Scope of the oxidation reaction. < J. Am. Chem. Soc. 1965, 81, 5670-5678] gave the desired peptides. Activated ketones containing a P2 residue other than asparagine were prepared using standard solution methods from alcohol 3 or by the new solid phase technique described below.
Schemes 3A and 3B: The synthesis of peptidyl trifluoromethyl ketone or α-ketoamide typically develops in solution, preparing a precursor alcohol and subjecting it to a final oxidation step. This oxidation is often problematic (especially when other oxidizable groups are present in the molecule) and sometimes limits the choice of pharmacophore to be incorporated into the inhibitor To take advantage of recent advances in robotic technologies and in the development of co-binary chemical techniques, an attempt was made to investigate a solid phase process for the synthesis of peptidyl trifluoromethyl ketone or a-ketoamide inhibitors. The goal was to develop a methodology that would give direct access to activated ketone functionality without the need to perform a final oxidation step. For this purpose, it was considered to use a semicarbazone ligand 103 (Scheme 3A) to serve as a reversible protective group for the ketone and as an anchor group for the polymeric support. A similar solid phase process has already been reported by Webb and collaborators for the preparation of peptidyl aldehydes (a) Murphy, A.M .; Dagnino, R .; Purity Jr. , L.V .; Trippe, A.J .; Sherman, S.L .; Lumpkin, R.H .; Tamura, S.Y .; Webb, T.R. J. Am. Ch em. Soc. 1992, 114, 3156; b) Webb, T.R. U.S. Patent 5,283,293; c) Webb, T.R. U.S. Patent 5,367,072). The process, however, comprises a final cutting step that is carried out without the formaldehyde requirement as described in the Webb et al. This allows for a greater variety of pharmacophores to be incorporated into the inhibitor.
The precursor for α-ketoamides (108) was prepared as follows: 23 108 Scheme 3 A 108 109 R: A.A. X * side chain: CF3 or C (O) NH-Bn oxalyl chloride, DMSO / CH2Cl2 des. Et3N, -78 ° C at 0 ° C resin i) 4N HCl / Dioxane then KgCOgac. (94%) f ii) 5 a-c, p-TsOH (cat.), Toluene reflux; iii) Pd / Ct H = (40 psi) / MeOH-EtOAc; BHA resin TBTU, HOBt, DIPEA / DMSO; v) 45% TFA / CH2Cl2- after 5% DIPEA / CH Cl A: divalent spacer group such as cyclohexyl, phenyl or benzyl The trifluoromethyl ketone or α-ketoamide (108) was oxidized by an oxidation of Swern to give the corresponding trifluoromethyl ketone and a-ketoamide in 66% yield.
With the necessary activated ketone available, it was sufficient to generate the desired icarbazone fraction 112 and to anchor it in a BHA resin. For this purpose, protected semicarbazide 110 was deprotected and neutralized. The resulting semicarbazide was then condensed in refluxing toluene, with activated ketone 109 under acid catalysis and azeotropic water removal, to give the trans icarbazone 112 in moderate yield (in the case of ketoamide, a mixture was obtained ci s / trans). The hydrogenolysis of the benzyl ester proceeded without problem to give the corresponding acid 113 in quantitative yield. The acid was then coupled to a BHA polystyrene resin by in-situ activation with TBTU followed by removal of the Boc protecting group to give the desired resin 103.
With the resin 103 available, the solid phase oligomerization was performed using standard protocols. The semicarbazone bond that is resistant to anhydrous and moderately basic acidic conditions, the amino acids protected with Boc and Fmoc could be used at any position of the peptide sequence. This versatility allowed an increased diversity in the building blocks to be incorporated in the final inhibitor. The amino acid coupling was made by their corresponding HOBt esters as shown in Scheme 3B. At the end of the synthesis, in the cases where the molecule incorporated acid-sensitive side chain protecting groups, they were removed by treatment with 75% F / CH 2 Cl 2 Scheme 3B 1- Boc-AA, coupling agent HOBt, DMF 2- 45% TFA / CH2C12 3- 5% DIPEA / CH2C12 4- Repetition of Step 1 as necessary The final cut of the polymer support was performed by refluxing the dried resin in a THF solution containing aqueous HCl and acetic acid at 65 ° C. To maximize yields, it was found necessary to filter the resin and repeat this protocol once again. In general, the resin cut was slightly slower for trifluoromethyl ketones derived from valine 212-217 (see Example 62) than for its alanine (201-207) or ethyl glycine (208-211) counterparts. This difference in hydrolysis rate could be compensated by making an extra cut for the valine derivatives. In the cases where a basic waste was presented in the sequence, a slightly higher concentration of HCl was used during the cut and a total of three cuts were required to ensure maximum yields. It has been found that the addition of formaldehyde to trap the semicarbazide released was superfluous. Not only did formaldehyde not provide any significant benefit as reflected by the total yield of the compound, but it also did not complicate the isolation of the desired product, particularly for sequences containing a free amino group.During the final cut, no interference of nucleophilic or oxidizable side chains such as those present in serine, methionine, tyrosine, histidine, lysine or aspartic acid was observed. The cleavage of inhibitors containing an asparagine residue adjacent to the trifluoromethyl group was more problematic. In this case, it was found necessary to use the N-trityl protected asparagine and deprotect the trifyl group in solution after the resin cut. The presence of an asparagine residue somewhere in the sequence did not, however, require any protection of the side chain.
The cutting conditions were quite light to be compatible with various acid sensitive protective groups such as N-Boc, ether O-t-Bu, ester O-t-Bu and ester O-Bn. During cutting, however, the methyl esters were hydrolysed to a degree of -50%. In most cases, treatment with 75% TFA before cutting the resin was sufficient to completely deprotect the acid-sensitive side chain protecting groups. However, in a few examples the O-t-Bu derivative of threonine and aspartic acid were also isolated indicating that the deprotection was not complete in these cases.
The a-ketoamides could also be synthesized by the same process (compounds 218 and 219 of example 62) and in similar total yields.
For its generality, this methodology is very suitable for an application in rapid advance optimization and in the generation of libraries with the purpose of identifying new inhibitors of trifluoromethyl ketone and a-ketoamide "de-serine proteases.
The protocols and yields of purified final products are reported in Examples 1 and 62 respectively.
Solution Chemistry Schemes 4, 5, and 6: Peptides containing activated ketones other than trifluoromethyl ketone could also be prepared by sequentially coupling a suitably protected amino acid with the required amino acids or peptide segment using standard solution methods. After the entire structure was established, oxidation of the resulting alcohol gave the desired compound. The preparation of several building blocks is shown in Schemes 4, 5 and 6.
Condensation of the Weinreb amide 10 with CF3CF2Li followed by reduction with NaBH4 gave the alcohol substituted with pentaf luoroethyl 11 (Scheme 4).
Eachema 4 OH BocHN OMe BocHN N CF2CF3 BocHN BocHN CbzHN 21: X = CH,.? = R = H, Ro = CHo The α, α-difluoroamide 13 was prepared from an Ultrasonic Reformatsky reaction (Thaisrivongs, S .; Country, PT; Kati, WM; Turner, SR; Thomasco, LM; Watt, W .; Desing and synthesis of potent and specific renin inhibitors containing diflurostatine, difluorstatone, and related analogues J. Med. Ch., 1986, 24, 2080-2087) between ethyl bromodifluoroacetate and Boc-alaninal followed by treatment with benzylamine. Benzothiazole 14 was obtained in a linear manner when 2-lithiobenzothiazole was added to this same aldehyde. The remaining benzoxazole derivatives are 16 to 21 were synthesized as shown for the amide 15. The reduction to the aldehyde by the action of LiAlH4 was followed by the formation of cyanohydrin, partial hydrolysis and cyclization using previously described procedures [(a) Ed ards, PD; Meyer, E.F. Jr.; Vij ayalakshmi, I .; Tuthill, P.A .; Andisik, D.A .; Gomes, B .; Strimpler, A. Design, synthesis, and kinetic evaluation of a unique class of elastase inhibitors, the peptidyl a-ketobenzoxazoles, and the X-ray crystal structure of the covalent complex between porcine pancreatic elastase and Ac-Ala-Pro-Val-2 -benzoxazole J. Am. Ch em. Soc. 1992, 1 1 4, 1854-1863. (b) Edwards, P.D .; Zottola, M.A .; Davis, M .; Williams, J .; Tuthill, P.A. Peptidyl a-ketoheterocyclic inhibitors of human neutrophil elastase. 3. In vi tro and in vi vo potency of a series of peptidyl a-ketobenzoxazoles. J. Med. Ch em. 1995, 38, 3972-3982. (c) Edwards, P.D .; Wolanin, D.J .; Andisik, D.W .; Davis, M.W. Peptidyl a-ketoheterocyclic inhibitors of human neutrophil elastase. 2. Effect of varying the heterocyclic ring on in tro tro potency. J. Med. Ch em. 1995, 38, 76-85. (d) Tsutsumi, S .; Okonogi, T .; Shibahara, A .; Ohuchi, S .; Hatsushiba, E .; Patchett, A. A .; Christensen, B.G. Synthesis and structure-act ivity relationships of peptidyl a-keto heterocycles as novel inhibitors of prolyl endopept idase. J. Med. Ch em. 1994, 37, 3492-3502].
Various α-ketoamide derivatives could also be prepared according to the alternative procedure depicted in Scheme 5.
Scheme 5 r- 24: R = OH 25 L 25: R = NR'R "Route a: A reaction of Henry between a glyoxylic acid and nitroethane gave 23 after reduction and adequate protection.The coupling of the amino acids required using the standard methods gave 24 after removal of the benzyl ester The incorporation of the appropriate amide function gave a series of alcohols which were readily oxidized to the desired ketones using the Dess-Martin reagent (Dess, DB; Martin, JC Readily accessible 12-1-5 oxidant for the conversion of primary and secondary alcohols to aldehydes and ketones, J. Org. Chem. 1983, 48, 4155-4156).
Route b: Alternatively, 23 could be hydrogenated to the corresponding acid and the appropriate Pl 'amine coupled. The coupling of the required amino acids using the standard methods gave 25. The alcohol 25 was easily oxidized to the desired ketones as before.
The preparation of the unnatural amino acids adamantil glycine and β, β-dimethyl aspartic acid is shown in Scheme 6. 32 C 33: R = H 34: R = Bn 29: R = adamantane 30: R = adamantane 31: R = adamantane 34: R = C (CH3) 2CO2Bn 35: R = C (CH3) 2CO2Bn 36: R = C (CH3) 2CO2Bn Thus oxazolidinone 29 was obtained from acid 27 using the previously described procedures (Gage, JR; Evans, DA Diastereoselective aldol condensation using a chiral oxazolidinone auxiliary: (2S *, 3S *) -3-hydroxy-3-phenyl-2-methylpro ? anoic acid, Org Syn. 1989, 68, 83-91). The formation of the enolate followed by treatment with TrisN3 (Evans, DA, Britton, TC, Ellman, JA, Dorow, RL The asymmetric synthesis of a-amino acids, Electrophilic azidation of chiral imide enolates, a practical approach to the synthesis of (R ) - and (S) -azido carboxylic acids, J. Am. Ch., Soc. 1990, 1 12, 4011-4030) gave the azide which was hydrolyzed to produce acid 31 in a linear fashion. The appropriate amino acid residues were then introduced into the carboxylate group of azido acid. The coverage of the term N was carried out using standard coupling methods after the reduction of the azide radical. Using a similar approach, the anhydride 32 was converted to the protected azido acid 36.
The following examples are provided to describe the invention in more detail. These examples, which establish the best form currently contemplated for carrying out the invention, are intended to illustrate and not to limit the invention.
Examples Unless otherwise indicated, the materials were obtained from commercial sources and used without further purification. The purity of each inhibitor was determined by HPLC, ^ H-NMR, and / or elemental analysis. The H-NMR spectra were obtained at 400 MHz on a Bruker AMX 400 spectrometer. The FAB mass spectrum was recorded on an Autospec, VG spectrometer. Column chromatography was performed either on silica gel (10-40 μm or 230-400 mesh ASTM, E. Merck) or by preparative HPLC using a preparative column Partisil 10 ODS-3, C18 (50 cm x 22 mm) . Analytical HPLC was performed in the following systems; System A: Vydac C18 analytical column, 10 μm (24 cm x 4.6 mm); mobile phase, acetonitrile / 0.06% trifluoroacetic acid (TFA) in water / 0.06% TFA; System B: Vydac C18 analytical column, 5 μm (15 cm x 4.6 mm); mobile phase, acetonitrile in 50 mM NaH2P04 at pH 4.4; System C: Vydac C8 analytical column, 10 μm (24 cm x 4.6 mm); mobile phase, acetonitrile in 20 mM Na2HP04 at pH 8.0; System D: C8 symmetric buckling analytical column, 10 μm (15 cm x 3.9 mm); mobile phase, acetonitrile in 20 mM Na2HP04 at pH 9.0; System E: Supelcosil C8 analytical column, 5 μm (15 cm x 4.6 mm); mobile phase, acetonitrile / 0.1% TFA in water / 0.1% TFA at pH 2.0.
Abbreviations or symbols used in the examples, or throughout the present specification, include Boc, but ioxycarbonyl tertiary; BOP: benzothiazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate; DA-Tbg, deamino-butylglycine-tertiary amino (3, 3-dimethyl-ilbutylbutanoic acid); DCC: N, N '-dicyclohexylcarbodimide; DIC, 2-dimethylaminoisoproyl chloride hydrochloride; DMF, N, N, -dimethylformamide; DMSO, dimethylsulfoxide; DTT, dithiothreitol; EDC: l-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride salt; Fmoc, 9-fluorenylmethyloxycarbonyl; HCMV, human cytomegalovirus; HOBt, hydrated 1-hydroxybenzotriazole; MONTH, 4-morpholinenenesulfonic acid; NMP, N-methylpyrrolidone; PCR, polymerase chain reaction; Ph, phenyl; PMSF, phenylmethylsulfonyl fluoride; QSAR, structure, activity relationship of the quantitative structure; Tbg, butylglicin terci a ria; tBu, tertiary butyl; TFA, trifluoroacetic acid; Trt, triphenylmethyl; TBTU, O- (benzotriazol-1-yl) -1,3,3,3-tetramethyluronium tetrafluoroborate; TCEP, tris (2-carboxyethyl) phosphine hydrochloride; TRIS, tris (hydroxymethyl) aminomethane.
Example 1. General procedure for peptide solid phase synthesis: 1- Protocol Boc / DIC / HOB.
The peptides were assembled on an ACT396 peptide synthesizer sold by Advanced Chemtech (Louisville, KT). Each reaction vessel was loaded with the appropriate resins 103 (0.25 mmol) and washed successively with 3.5 mL portions of CH 2 Cl 2 (2 X), MeOH (2 X) and CH 2 Cl 2 (2 X). The amino acids were coupled as their activated HOBt esters, using 4.8 equivalents of the reagents as follows: A 0.5 M solution of an amino acid mixture protected with Fmoc and HOBt in DMF (2.4 mL, 1.2 mmol of each) was added to the deprotected resin, followed by the addition of a solution of 0.5 M DIC in CH2Cl2 (2.4 mL, 1.2 mmol). The reaction vessel was stirred for 3.5 h. The reaction vessel was drained and the remaining resin was washed twice with 5 mL of CH2C12- Fresh portions of reagent solutions were added and the coupling step was repeated for 3.5 h. After coupling, the resin was washed successively with 5 mL portions of GH 2 Cl 2 (2 X), MeOH (2 X) and CH 2 Cl 2 (2 X). The amino Boc protecting groups were removed with a solution of 45% TFA in CH2Cl2 (4 mL for 25 min) and washed as before. After the last coupling, an additional wash step was made with 5 L of CH 2 Cl 2 (3 X) and the resin was dried in vacuo. 2- Protocol F oc / TBTU / HOBt.
The peptides were assembled as before except for the coupling which was done as follows: The resin was suspended in NMP (0.35 mL) and treated with a 0.5 M solution of an amino acid mixture protected with Fmoc and HOBt in NMP (1.8 mL, 0.9 mmol of each), a 0.5 M solution of TBTU in DMF (1.8 L, 0.9 mmol) and a 1.0 M solution of DIPEA in NMP (1.8 mL, 1.8 mmol). The reaction vessel was stirred for 1.25 h, drained and the remaining resin was washed twice with 3.5 mL of DMF. Fresh portions of reagent solutions were added and the coupling step was repeated for 1.25 h. Deprotection of the Fmoc group was done by treating the resin with a 25% solution of piperidine in DMF for 25 minutes.
Protocol Boc / TBTU / HOBt.
The peptides were assembled on a peptide synthesizer COUPLER ™ 250 C (VEGA Biotechnologies) or on an ACT 90 (Advanced ChemTech). The reaction vessel was charged with the appropriate resins 103 (0.25 mmol) which were washed successively with 15 mL portions of CH 2 Cl 2 (2 X), MeOH (2 X) and CH 2 Cl 2 (2 X). The amino acids were coupled as their activated HOBt esters, using 3 equivalents of the reagents as follows: The resin was suspended in DMF (15 L) and treated with the amino acid protected with Boc (0.75 mmol), HOBt hydrated (0.75 mmol), DIPEA (1.5 mmol, 0.26) and TBTU (0.75 mmol, 241 mg). The reaction vessel was stirred for 1 h and the termination of the coupling was monitored by means of the Kaiser test. In the case of incomplete links, the reaction vessel was drained and the resin was washed twice with 15 mL of CH2CL2. The fresh reagents were added and the coupling step repeated for an extra hour. The reaction vessel was drained and the resin washed as before. The Boc protecting group was removed by successive treatment (5 min. After 20 min.) With 15 mL of a 45% solution of TFA in CH2C12- The resin was washed with CH2CL2 (2 X), 5% DIPEA in CH2CL2 ( 1 min, then 5 min.), CH 2 Cl 2 (2 X), MeOH (2 X) and CH 2 Cl 2 (2 X).
General Procedure for the cutting of peptidyl-activated ketone from the solid support (according to Scheme 3B).
The dried resin (~ 800 mg) was suspended in THF (9 mL), H20 (0.50 mL), AcOH (0.25 mL) and aq. 1M (0.10 mL) and heated in a bomb at 65 ° C for four hours. The solution was cooled, filtered and treated as before once more. In the case where the sequence contained a basic residue such as lysine or histidine, 0.05 mL extra of 1 M HCl was used and the procedure was repeated three times. All mother liquors were combined, THF was concentrated in vacuo and the residue was purified on reverse phase HPLC (Whatman HPLC column, 22.0 mm * 500 mm, Partisil 10 ODS-3 M / 20-50, particle size 10 μm, solvents: A = 0.06% TFA / H20, B = 75% CH3CN-25% H20 containing 0.06% TFA; gradient 0 to ~ 50% of B in 60 min). The desired ketones activated with peptidyl 201-219 were isolated in yields reported in Example 62.
Example 2. Alternative synthesis of α-ketoamides (according to scheme 5). Preparation of benzyl amide of acid 3-. { 2- [2- (3, 3-dimethyl-butyrylamino) -3,3-dimethyl-butyrylamino] -3-dimethylcarbamoyl-propionylamino} -2 -oxo-butyric (76, Table 6).
Preparation of a-hydroxy ester 23. To a solution of nitroethane (4.0 g, 53 mmol) in ethanol (15 mL) was added aqueous NaOH (68 mL of 2N solution, 136 mmol). To this stirred solution, glyoxylic acid (5.9 g, 64 mmol) was added rapidly. The solution was stirred 15 h and then acidified with 10% aqueous HCl (pH 2) and the aqueous phase was saturated with NaCl before extraction with EtOAc (3 x 150 mL). The organic phase was dried with (MgSO), filtered and concentrated to give 8.1 g of a viscous yellow oil. This crude material was dissolved in ethanol (50 mL) containing Et3N (18 L, 119 mmol) and treated with di-tert-butyl bicarbonate (12.2 g, 56 mmol) and Raney nickel (3 g) which had been added. Wash with water immediately before use. Hydrogenation at 45 p.s.i for 20 h provided after filtration and concentration through Celite, the crude acid (11.1 g). A portion of the crude acid (3.07 g, 14 mmol) was dissolved in DMF (30 L) and treated with anhydrous K2CO3 (4.3 g, 30.8 mmol) and benzyl bromide (2.5 mL, 21 mmol). After stirring for 3 h at room temperature, the DMF was removed under reduced pressure and the residue was dissolved in EtOAc (150 mL) and washed with water (100 mL) and brine (80 L). The organic phase was dried with MgSO, filtered and concentrated. The crude yellow oil (4.3 g) was purified by flash chromatography on silica gel (230-400 mesh), eluting with 33% EtOAc in hexane to provide pure benzyl ester 23 (1.8 g, 42% nitroethane). HPLC (system C) 99%, (system D) 97%; IR (KBr)? 3422, 3361, 1740, 1684 cm "1; XH-NMR (400 MHz, CDC13) d 7.36 (s, 5H), 5.27 (d, J == 12.1 Hz, ÍH), 5.19 (d, J = 12.1 Hz, ÍH), 4.82 (m, ÍH), 4.36 and 4.35 (2 xd, J = 5.7 and 5.4 Hz, ÍH), 4.11 (m, 1H), 3.10 (m, ÍH), 1.43 (s, 9H), 0.97 ( d, J = 7.0 Hz, 3H); FAB MS m / z: 310 (MH +), 210 (M-100); HRMS cale for C? 6H24N05 (MH +) 310.1654, Found: 310.1644; Anal (Ci6H23N05) C, H , N.
Synthesis of α-keto acid 24. The tert-butyloxycarbonyl (Boc) group of ester 23 (4.0 g, 12.9 mmol) was removed using 4 N HCl / dioxane (30 mL) for 45 min at 0 ° C. The hydrochloride salt was obtained by concentration and coevaporation with toluene (15 mL). The HCl salt (12.9 mmol) was combined with EDC (2.6 g, 13.6 mmol, 1.1 equiv.), HOBt (1.8 g, 13.6 mmol, 1.1 equiv.) And Boc-Asn (NMe2) -OH (3.4 g, 12.9 mmol, 1.1 equiv.) In DMF (50 mL) under a nitrogen atmosphere. The solution was cooled to 0 ° C (ice bath) before iPr2NEt (7.9 mL, 45.3 mmol, 3.5 equiv.) Was added. Then the solution was stirred at room temperature for 16 h. The reaction mixture was partitioned between EtOAc (250 mL) and aqueous sat. NaHCO 3. (150 mL). The organic phase was washed with aq. at 5% (150 L) and finally brine (150 mL). Drying (MgSO) was followed by filtration and concentration to give 6.0 g of crude material. In most cases the raw material was suitable for subsequent couplings without purification. After the final coupling, the α-hydroxy benzyl ester peptides were purified by flash chromatography. The acid 24 was obtained after the benzyl ester (1.10 g, 2.0 mmol) by hydrogenation with 10% Pd / C (55 mg) in ethanol (30 L) at atmospheric pressure over the course of a few hours to provide after filtration through a pad of Celite a white solid (0.95 g, 100% yield). HPLC (system A) 100%, (system C) 100%; IR (KBr)? 3316, 1727, 1642 cpr1; ^ -RM (400 MHz, CDC13), mixture of 4 diastereomers, d 8.06 and 8.01 (2 xd, J = 7.3 and 8.6 Hz, ÍH), 7.87, 7.79, 7.70 and 7.54 (4 xd, J = 8.6, 8.6, 8.9 and 8.6 Hz, ÍH), 7.09 and 7.03 (2 xd, J = 7.9 and 8.6 Hz, 0.5H), 6.72 (m, 0.5H), 6.52 (m, 0.25), 6.34 and 6.29 (2 xd, J = 7.6 and 7.3 Hz, 0.75H), 6.10-5.4 (br s, ÍH), 4.99-4.88 (m, 0.5H), 4.87-4.78 (m, 0.5H), 4.66-4.37 (m, 2H), 4.33- 4.09 (m, ÍH), 3.30-3.15 (, 0.3H), 3.05-2.85 (m, 6.7 H), 2.75-2.65 and 2.60-2.50 (m, 1H), 2.25-2.10 (, 2H), 1.28-1.19 (, 3H), 1.10-0.97 (m, 18H); 13 C-NMR (100.6 MHz, CDC13) d 174.9, 173.5, 173.1, 173.0, 171.5, 171.0, 170.9, 170.8, 170.7, 170.5, 170.2, 73.03, 72.74, 60.9, 60.67, 50.3, 50.1, 49.5, 49.4, 48.1, 47.9, 47.7, 37.56, 35.9, 35.8, 35.7, 34.7, 34.4, 34.3, 33.8, 31.1, 29.9, 29.8, 26.9, 26.8, 26.7, 17.4, 17.2; FAB MS m / z: 473 (MH +), 495 (M + 23); HRMS for C22H4? N407 (MH +) 473.2975, found: 473.2990; Anal (C22H40N4O7) C, H, N.
The coupling of the Pi 'residue was performed using the above general coupling protocol with the appropriate terminal amine (1.2 equiv.). The final oxidation step was developed by treatment of the prerequired a-hydroxy amide (62 mg, 0.11 mmol) with 2 equivalents of Dess-Martin periodinnan (94 mg, 0.22 mmol) in DMF (1 mL) for 4 h. The addition of 10% sodium thiosulfate (5 mL) and sat. NaHCO 3. (5 mL) with stirring (15 min) was followed by extraction with EtOAc (3 x 10 L) to give the desired a-ketoamide. The final purification was developed using preparative HPLC to provide 76 after lyophilization, (51 mg, 82% yield) as a white solid. HPLC (system C) 100%, (system D) 96.1%; IR (KBr)? 3316, 1641, 1529 cm'1; XH-NMR (400 MHz, DMSO-d6), 1: 1 mixture of diastereomers in Px, d 9.21-9.15 (m, HH), 8.14 and 8.09 (2 xd, 7.3 and 7.6 Hz, HH), 8.03 and 7.97 ( 2 xd, J = 6.4 and 5.7 Hz, ÍH), 7.60 (d, J = 8.3, ÍH), 7.35-7.17 (m, 5H), 5.02-4.88 (m, ÍH), 4.64-4.49 (m, ÍH) , 4.39-4.23 (m, 2H), 4.13 and 4.12 (2 xd, J = 8.6 and 8.6 Hz, ÍH), 2.92 and 2.91 (2 xs, 3H), 2.79 and 2.78 (2 xs, 3H), 2.74-2.54 (m, 2H), 2.19 (br d, J = 12.4 Hz, ÍH), 2.03 and 2.02 (2 xd, J = 12.4 and 12.7 Hz, ÍH), 1.25 and 1.23 (2 xd, J = 7.3 and 7.0 Hz, 3H), 0.94 and 0.91 (2 xs, 18H); FAB MS m / z: 560 (MH +), 582 (M + 23); HRMS cale for C29H46N5? 6 (MH +) 560.3448, found: 560.3426.
Example 3. Alternative preparation of Fmos-Asp (Rink resin) - aminotrifluoromethyl alcohol (5) (according to scheme 2; R: Me). 4- (er-butoxycarbonylamino) -1,1, 1-trifluorobutan-2-ol (3) . This compound was prepared according to an analogous procedure of the literature for the preparation of valine analogue (Skiles, JW; Fuchs, V .; Miao, C; Sorcek, R .; Grozinger, KG; Mauldin, SC; Vitous, J .; Mui, PW, Jacober, S., Chow, G., Matteo, M., Skoog, M., Weldon, ST, Possanza, G., Keirns, J., Letts, G., Rosenthal, A. Inhibition. of human leukocyte elastase (HLE) by N-substituted peptidyl trifluoromethyl ketones, J. Med. Chem. 1992, 35, 641-662). IR (KBr)? 3313 (br), 1681, 1527 cm "1; XH-NMR (400 MHz, DMSO-d6), 1: 1 mixture of diastereomers, d 6.84 and 6.43 (d, J = 8.4 Hz and 8.9 Hz, ÍH), 6.32 and 6.24 (d, J = 6.9 and 7.4 Hz, ÍH), 3.93-3.81 (m, 1.5H), 3.70 (quint, J = 7.3 Hz, 0.5H), 1.373 and 1.371 (s, 9H), 1.10 and 1.07 (d, J = 7.4 and 6.4 Hz, 3H); FAB MS m / z: 244 (MH +); Anal (C9H? 6F3N03) C, H, N.
Fmoc-Asp (Ot-Bu) -aminotrifluoromethyl alcohol (4). This compound was prepared in solution by coupling Fmoc-Asp (Ot-Bu) -OH and the aminotrifluoromethyl alcohol derived from alanine. HPLC (system A) 98%, (system B) 99%; IR (KBr)? 3300, 1721, 1697, 1661, 1540 crn "1; 1 H-NMR (400 MHz, DMSO-d 6) d 7.99-7.93 (m, HH), 7.89 (d, J = 7.3 Hz, 2H), 7.70 (m, 2H), 7.55 (dd, l = 15.9 Hz , J "2 = 8.6 Hz, 1H), 7.45-7.39 (m, 2H), 7.35-7.29 (m, 2H), 6.41 (d, J = 7.0 Hz, ÍH), 4.40-4.19 (m, 4H), 4.06-3.88 (m, 2H), 2.66-2.58 (m, ÍH), 2.44-2.40 (m, 1H), 1.37 (s, 9H), 1.08 (d, J = 6.7 Hz, 3H), FAB MS m / z: 537 (MH +); HRMS cale for C 27 H 32 F 3 N 206 (MH +) 537.2212, found: 537.2229.
The above compound was deprotected with 40% TFA in CH2C12 and coupled in a Rink resin with DCC / HOBt to provide (5).
Example 4. Preparation of other activated ketones (according to scheme 4). 4- (ßr-Butoxycarbonylamino) -1, 1,1,2,2-pentafluoropentan-3-ol (11). To a dry 500 mL round bottom flask was added anhydrous Et20 (100 mL) and a 1.5 M solution of MeLi-LiBr in Et20 (100 mL, 150 mmol). This solution was subsequently cooled to -78 ° C. A second flask was cooled to -78 ° C and charged with Et20 (100 L) and CF3CF2I (44.7 g, 182 mmol). The content of this flask was then added via cannula for 15 min to the suspension of MeLi-LiBr. The resulting solution was stirred for 30 min at -78 ° C before the Weinreb amide 10 (10.6, 45.5 mmol) was added in one portion. The reaction was stirred at -78 ° C for 90 min and then allowed to warm to -30 ° C for 2 h. The reaction was quenched by the addition of sat. NH 4 Cl. (125 m). The organic phase was washed with H20 (2 x 50 L), dried over Na2SO4, filtered and concentrated to give an orange oil. The oil was redissolved in 20% MeOH / THF (100 m) and transferred to a 500 mL round bottom flask. The solution was cooled to 0 ° C before NaBH 4 (1.9 g, 50.1 mmol) was added portion by portion for 5 min (Caution! Can present foam). The reaction was subsequently stirred for 1 h at 0 ° C. Et20 (200 mL) was added followed by 10% citric acid (100 mL). The aqueous layer was extracted with Et20 (3 x 50 mL). The combined organic extracts were washed with NaHCO 3 (1 x 50 mL), brine (1 x 50 mL), dried with Na 2 SO 4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (20% ethyl acetate in hexanes) to give a colorless oil 11, 12.3 g (92%). HPLC (system D) 100%, IR (KBr)? 3368, 2987, 1687 cpr1; lH-NMR (400 MHz, DMSO-de) 25: 1 mixture of diastereomers, d 6.87 (d, J = 8.0, 1.5H), 6.42 (d, J = 8.9 Hz, 0.5H), 6.32 and 6.25 (2 xd , J = 8.0 and 8.3 Hz, ÍH), 4.08-3.78 (, 2H), 1.37 (s, 9H), 1.13 and 1.08 (2 xd, J = 7.0 and 6.7 Hz, 3H); FAB MS m / z: 294 (MH +); HRMS for C? 0H? 7F5NO3 (MH +) 294.1129, found: 294.1138. 4- (Er-butoxycarbonylamino) -2,2-difluoro-3-hydroxy- (4S) -benzyl pentanoate (13). This material was prepared using a modification of the previously described procedure (Thaisrivongs, S .; País, P.T .; Kati, W.M .; Turner, S.R .; Thomasco, L.M .; Watt, W .; Design and synthesis of potent and specific renin inhibitors containing diflurostatine, difluorstatone, and related analogues. J. Med. Ch em. 1986, 24, 2080-2087). Thus amide 10 (14.9 g, 64.3 mmol) was dissolved in THF (230 mL) at 0 ° C. LiAlH4 (4.90 g, 129 mmol) was added in several portions over a period of 20 min, and the suspension was then stirred for 2 h at 0 ° C. This suspension was transferred via cannula in 500 mL of 10% aqueous citric acid and stirred for 1 h. The mixture was extracted with Et20 (3x) and the combined organic phases were washed with water, brine, dried (MgSO4), filtered and concentrated to give the corresponding aldehyde as a white solid (10.7 g, 97%). ^ -RM (CDC13) d 9.57 (s, ÍH), 5.09 (br, 1H), 4.23 (br, ÍH), 1.46 (s, 9H), 1.34 (d, J = 7.3 Hz, 3H). Zinc powder (16.2 g, 24.8 mmol) was placed in THF (40 mL) and sonicated 30 min. A solution of the aldehyde (10.7 g, 62 mmol) and ethyl bromodifluoroacetate (20 g, 99 mmol) was added for 30 min using a needle pump while sonicating. Sonication was continued for 1.5 h before the suspension was poured into 500 L of 10% aqueous citric acid and extracted with EtOAc (3x). The combined organic extracts were washed with water, brine, dried (MgSO), filtered and concentrated in vacuo. The oil obtained contained 20% of the initial aldehyde but was used without further purification. The hydroxy ester (2.20 g, 7.40 mmol), benzylamine (3.96 g, 37.0 mmol) and i-Pr2NEt (4.76 g, 37.0 mmol) was heated in refluxing ethanol for 18 h. The solution was concentrated to dryness, taken up in EtOAc and washed with 1 N HCl, brine, dried (MgSO 4), filtered and concentrated in vacuo to give a yellow oil. This material was purified by flash chromatography using TLC grade silica gel to give 13 as a white solid (1.17g, 44% in 2 steps). HPLC (system B) 100%, (system C) 99%; IR (KBr)? 3344, 2979, 1684 CITG1; 1H-NMR (CDC13) 4: 1 isomer mixture d 7.20 (m, 5H), 6.90 (br s, ÍH), 4.75 (m, 1H), 4.45 (m, 2H), 4.08 (br s, ÍH), 3.88 (m, 2H), 1.32 (s, 9H), 1.16 (d, 3H); 13 C-NMR (100.6 MHz, DMS0-d6) d 164.58, 164.30, 164.02, 155.61, 139.31, 129.11, 127.97, 127.72, 119.99, 117.47, 114.88, 78.59, 71.91, 71.65, 71.44, 46.28, 43.04, 29.10, 19.35; FAB MS m / z: 359 (MH +); HRMS for C17H2SF2N204 (MH +) 359.1782, found: 359.1768; Anal (C17H24F2N204) C, H, N. (2S) -2- (er-butoxycarbonylamino) -1-benzo [d] [1, 3] thiazol-2-yl-l-propanol (14). This compound was prepared from the methyl ester 12 using the procedure previously described (Tsutsumi, S., Okonogi, T., Shibahara, A., Ohuchi, S., Hatsushiba, E., Patchett, AA, Christensen, BG Synthesis and structure-activities of peptidyl a-keto heterocycles as novel inhibitors of prolyl endopeptidase, J. Med. Ch., 1994, 37, 3492-3502). Thus a solution of 12 (28.1 g, 132 mmol) in THF (50 mL) was added dropwise to a suspension of LiAlH4 (3.76 g, 396 mmol) in THF (200 mL) at 0 ° C. After the addition was complete the mixture was stirred at room temperature for one hour. Celite (34 g) was then added followed by the careful addition of water (34 mL), 2N NaOH (34 mL) and water (100 mL) and stirring was continued for one hour. The resulting white suspension was filtered, and the filter cake was washed with EtOAc. The desired alcohol was obtained as a colorless oil (21.07 g, 91%). XH-NMR (CDC13) d 4.66 (br s, 1H), 3.77 (br s, ÍH), 3.68-3.61 (m, ÍH), 3.53-3.47 (m, ÍH), 2.65 (br s, ÍH), 1.45 (s, 9H), 1.14 (d, J = 6.7 Hz, 3H). To a solution of this alcohol (3.25 g, 18.5 mmol) and Et3N (7.75 mL, 55.6 mmol) in anhydrous CH2C12 (60 mL) and DMSO (28 L) at 0 ° C was added S03-pi (8.85 g, 55.6 mmol). ) in small portions. The solution was then stirred at room temperature for 1.5 h before it was poured into ice water and extracted three times with CH2C12. The combined organic extracts were dried (MgSO4), filtered and concentrated to give an oil which was purified by flash chromatography to give the desired aldehyde (2.34 g, 73%) which was used immediately. XH-NMR (CDC13) d 9.57 (s, 1H), 5.09 (br, ÍH), 4.23 (br, 1H), 1.46 (s, 9H), 1.34 (d, J-7.3 Hz, 3H). To a solution of benzothiazole (4.43 L, 40.5 mmol) in THF (100 mL) at -78 ° C was added nBuLi (26.5 mL of a 1.4M solution in hexanes, 37.15 mmol). After stirring for 30 min, a solution of the above aldehyde (2.34 g, 13.51 mmol) in THF was added. The solution was stirred for 72 min before being quenched by the addition of saturated NH 4 Cl. Extraction with EtOAc was followed by a wash with brine and dried with MgSO4. Flash chromatography gave the desired product as an orange oil. HPLC (system A) 100%, (system D, pH 7.4) 100%; IR (KBr)? 3272, 1713 cra "1; XH-NMR (400 MHz, DMS0-d6), 6: 1 mixture of diastereomers in Pj, d 8.07-8.05 (m, ÍH), 7.96-7.92 (m, ÍH), 7.50- 7.45 (m, ÍH), 7.41-7.37 (m, ÍH), 6.81 and 6.44 (2 xd, J = 8.6 Hz, ÍH), 6.47 (d, J = 5.4 Hz, ÍH), 4.92-4.90 (, 1H) , 4.03-3.96 (m, 1H), 1.32 and 1.29 (2 xs, 9H), 1.08 and 0.98 (2 xd, J = 6.7 and 6.7 Hz, 3H); FAB MS m / z: 309 (MH +), HRMS cale for C1 SH21N203S (MH +) 3 0 9, 12 7 3, found: 30 9, 128 3 (2S) -2- (benzyloxycarbonylamino) -1-benzo [d] [1, 3] oxazol-2-yl-l-propanol (16). This compound was prepared from 15 and 2-aminophenol using the procedure previously described ((a) Edwards, PD; Meyer, EF Jr.; Vij ayalakshmi, I; Tuthill, PA; Andisik, DA; Gomes, B .; Strimpler, A. Design, synthesis, and kinetic evaluation of a unique class of elastase inhibitors, the peptidyl a-ketobenzoxazoles, and the X-ray crystal structure of the covalent complex between porcine pancreatic elastase and Ac-Ala-Pro-Val-2 -benzoxazole, J. Am. Chem. Soc. 1992, 114, 1854-1863). To a solution of N-benzyloxycarbonyl- (S) -alanine (20.0 g, 89.7 mmol) in CH2C12 (200 mL) at 0 ° C, 1-1'-carbonyldiimidazole (18.2 g, 115.7 mmolj was added after 30 min. stirring at 0 ° C, Et 3 N (16.1 mL, 115.7 mmol) was added followed by the addition of O, N-dimethylhydroxylamine hydrochloride (11.3 g, 115.7 mmol) The mixture was stirred 1 h at 0 ° C and then at room temperature At room temperature for 4 h, CH2C12 was added and the organic phase was washed twice with 10% aqueous HCl, saturated NaHC03 and brine and dried with MgSO.The removal of the solvent in vacuo gave the amide 15 (24.2 g) which was used without further purification. ^ -RM (CDC13) d 7.4-7.3 (m, 5H), 5.65-5.55 (m, ÍH), 5.15-5.05 (m, 2H), 4.82-4.74 (, ÍH), 3.77 (s, 3H), 3.21 (s, 3H), 1.34 (d, J = 6.9 Hz, 3H). This compound was dissolved in THF (350 mL) at 0 ° C. A 1.0 M solution of LiAlH4 in THF (110 mL, 110 mmol) was added dropwise over 30 min. Stirring was continued at room temperature for 2 h. The mixture was then cooled to 0 ° C and a solution of KHS04 (22.4 g) in water (250 mL) was carefully added. After stirring at 0 ° C for one hour, the solution was extracted with ether and washed twice with 10% aqueous HCl, twice with saturated NaHCO 3 and once with brine. The organic phase was dried (MgSO4), filtered and concentrated in vacuo to give the desired aldehyde (19.4 g) which was immediately dissolved in CH2C12 (350 L) and cooled to 0 ° C. A solution of NaHS03 (55.9 g, 540 mmol) in water (150 mL) was introduced and the resulting mixture was stirred for one hour. NaCN (25.0 g, 511 mmol) was then added and the stirring was continued overnight. The suspension was diluted with EtOAc (250 mL) and hexanes (250 mL) and the layers were separated. Washing with water and brine was followed by drying with MgSO4 to provide the desired cyanohydrin (18.26 g, 83%) which was dissolved in benzene (350 mL) and stored at -20 ° C. AcCl (53.5 mL, 752 mmol) was added to a mixture of ethanol (47.1 mL, 803 mmol) and CHC13 (50 mL) at 0 ° C. After stirring at 0 ° C for 30 min. a solution of the above cyanohydrin (5.87 g, 25.1 mmol) in CHC13 (50 L) was added dropwise and the stirring was continued for an additional 2 h. The mixture was then concentrated in vacuo and taken up in ethanol (60 mL). The solution was refluxed in the presence of 2-aminophenol (3.01 g) throughout the night. The ethanol was removed and the residue was taken up in EtOAc, washed twice with % NaOH, 10% HCl, NaHCO 3 and brine and dried (MgSO 4). Flash chromatography afforded the desired compound 16 as an orange syrup (4.81 g, 59%) which was used without further purification. An analytical sample was obtained by recrystallization with 30% EtOAc in hexanes. HPLC (system A) 99%, (system D) 99%: IR (KBr)? 1692 citr1; XH-NMR (400 MHz, DMS0-d6), 1: 1 mixture of diastereomers in Px, d 7.75-7.68 (m, 2H), 7.42-7.19 (m, 8H), 6.22 and 6.10 (2 xd, J = 6.0 and 5.4 Hz, ÍH), 5.03-4.71 (m, 3H), 4.14-4.01 (m, 1H), 1.20 and 1.11 (2 xd, J = 6.7 and 7.0 Hz, 3H); FAB MS m / z: 327 (MH +); HRMS cale for d8HlsN204 (MH +) 327.1345, found: 327.1355; Anal (C18H18N204) C, H, N. (2S) -2- (benzyloxycarbonylamino) -1- (oxazolo [4, 5, b] pyridin-2-yl) -1-propanol (17). This material was prepared as a 1: 1 mixture of isomers in 12% yield of the previous cyanohydrin (978 mg, 4.18 mmol) and 2-amino-3-hydroxypiiridine (505 mg, 4.60 mmol) using the procedure described above for compound 16. An analytical sample was obtained by recrystallization with EtOAc in hexanes (an isomer). MP: 159-161 ° C; IR (KBr)? 1719, 1699 cm "1; XH-NMR (400 MHz, DMSO-d6) d 8.53 (dd, J = 4.8, 1.2 Hz, ÍH), 8.18 (d, J = 7.8 Hz, ÍH), 7.44 (dd, J = 8.1, 5.1 Hz, 1H), 7.39-7.08 (m, 6H), 6.33 (d, J = 6.0 Hz, ÍH), 5.00-4.82 (m, 2H), 4.74 (m, ÍH), 4.10-4.00 ( m, ÍH), 1.21 (d, J = 6.7 Hz, 3H), 13C-NMR (100.6 MHz, DMS0-d6) d 169.34, 155.32, 154.73, 146.08, 142.32, 136.99, 128.21, 127.59, 127.37, 120. 57, 119.04, 69.99, 64.99, 49.91, 16.31; FAB MS m / z: 328 (MH +); HRMS cale for C 17 H 18 N 304 (MH +) 328.1297, found: 328.1286; Anal (C17H17N304) C, H, N. (2S) -2- (benzyloxycarbonylamino) -1- (4-methylbenzo [d] [1,3] oxazol-2-yl) -1-propanol (18). This material was prepared as a 1: 1 mixture of isomers in 35% yield of the previous cyanohydrin (707 mg, 3.02 mmol) and 2-amino-m-cresol (409 mg, 3.32 mmol) using the procedure described above for compound 16. An analytical sample was obtained by recrystallization with EtOAc in hexanes (1.3: 1 mixture of isomers ). mp: 98 ° C; IR (KBr)? 1701, 1690 crn-1; iH-NMR (400 MHz, CDC13) 5 7.38-7.20 (m, 7H), 7.15- 7. 10 (m, 1H), 5.39 and 5.29 (2 xd, J = 7.9 and 5.4 Hz, ÍH), 5.15-4.90 (m, 3H), 4.40 and 4.23 (2 x br s, 2H), 2.58 (s, 3H) ), 1.34 and 1.14 (2 xd, J = 6.7 and 7.0 Hz, 3H); Í3c-NMR (100.6 MHz, CDCI3) d 164.82, 164.22, 156. 28, 156.12, 150.72, 150.65, 139.40, 136.27, 130. 61, 130.43, 128.51, 128.40, 128.14, 128.01, 127. 91, 125.20, 125.14, 125.04, 124.98, 108.20, 108. 13, 71.02, 70.62, 66.96, 66.77, 50.45, 50.27, 17.35, 16.39, 12.24; FAB MS m / z: 341 (MH +); HRMS for C19H21N2O4 (MH +) 341.1501, found: 341.1490; Anal (C? QH20N2 ° 4), H, N. (2S) -2- (benzyloxycarbonylamino) -1- (5-methylbenzo [d] [1,3] oxazol-2-yl) -1-propanol (19). This material was prepared as a 1: 1 mixture of isomers in 53% yield of the above cyanohydrin (1.10 g, 4.70 mmol) and 2-amino-p-cresol (636 mg, 5.17 mmol) using the procedure described above for the compound 16. An analytical sample was obtained by recrystallization with EtOAc in hexanes (7: 1 mixture of isomers). mp: 134-135 ° C; IR (KBr)? 1718, 1691 cm "1; ifi-RM (400 MHz, CDCI3) d 7.47-7.09 (m, 8H), 5.47 (d, J = 8.6 Hz, ÍH), 5.12-4.87 (m, 4H), 4.54-4.30 (m, ÍH), 2.44 (s, 3H), 1.32 and 1.13 (2 x d, J = 6.7 and 6.7 Hz, 3H); 13 C-NMR (100.6 MHz, CDCl 3) d 165. 90, 165.19, 156.09, 149.04, 140.24, 136.28, 134.38, 128.34, 128.09, 127.91, 126.32, 119.87, 119.69, 110.23, 70.84, 70.57, 66.92, 66.67, 50.48, 50. 29, 21.34, 17.24, 15.32; FAB MS m / z: 341 (MH +); HRMS cale for C19H2i 2? (MH +) 341.1501, found: 341.1490; Anal (C19H20N2O4) C, H, N. (2S) -2- (benzyloxycarbonylamino) -1- (6-methylbenzo [d] [1,3] oxazol-2-yl) -1-propanol (20). This material was prepared as a 1: 1 mixture of isomers in 71% yield of the previous cyanohydrin (1.44 g, 6.15 mmol) and 6-amino-m-cresol (832 mg, 6.77 mmol) using the procedure described above for compound 16. An analytical sample was obtained by recrystallization with EtOAc in hexanes (8: 1 mixture). isomers), mp: 108-109 ° C; IR (KBr)? 1692 cm -1 1 H H - NMR (400 MHz, CDCl 3) d 7.54 (d, J = 8.3 Hz, ÍH), 7.36-7.10 (m, 7H), 5.40 and 5.32 (d, J = 8.9 and 8.9 Hz, 1H), 5.14-4.85 (m, 3H), 4.42-4.28 (m, 2H), 2.47 (s, 3H), 1.33 and 1.14 (2 xd, J = 6.7 and 6.7 Hz, 3H); 13 C-NMR (100.6 MHz, CDCl 3) d 165.08, 156.11, 151.24, 137.95, 136.28, 135.76, 128.52, 128.16, 127.99, 127.90, 125.89, 125.79, 119.38, 119.20, 111.09, 70.99, 70.69, 66.99, 66.76, 50.55 , 50.26, 21.70, 17. 29, 15.38; FAB MS m / z: 341 (MH +); HRMS cale for C19H21N2 ° 4 (H +) 341.1501, found: 341.1490; Anal (C19H2? N2? 4) C, H, N. (2S) -2- (benzyloxycarbonylamino) -1- (7-methyl bßnzo [d] [1,3] oxazol-2-yl) -1-propanol (21). This material was prepared as a 1: 1 mixture of isomers in 57% yield of the previous cyanohydrin (609 mg, 2.60 mmol) and 6-amino-o-cresol (330 mg, 2.60 mmol) using the procedure described above for compound 16. An analytical sample was obtained (1.2: 1 mixture of isomers) by flash chromatography (30% EtOAc in hexanes). oil; IR (net)? 1705 was "1; iH-NMR (400 MHz, CDCI3) d 7.50 (, 1H), 7.35-7.10 (m, 7H), 5.50 and 5.38 (2 xd, J = 8.9 and 8.3 Hz, HI), 5.15-4.90 (m, 3H), 4.41 (s, 1H), 2.36 (s, 3H), 1. 35 and 1.14 (2 x d, J = 6.7 and 7.0 Hz, 3H); 13 C-NMR (100.6 MHz, CDCl 3) d 165.41, 164.85, 156.29, 156.12, 150. 14, 139.67, 136.28, 128.50, 128.39, 128.13, 127.97, 127.84, 126".36, 126.31, 124.59, 124.51, 121.54, 117.28, 117.12, 70.99, 70.56, 66.96, 66.71, 50.42, 50.25, 17.41, 15.26, 15.08; FAB MS m / z: 341 (MH +); HRMS cale for C19H21N2O4 (MH +) 341.1501, found: 341.1490; Anal (Ci9H20N2 ° 4) c 'H' N- Example 5. Preparation of non-natural amino acids (according to scheme 6). (4S) -3- [2- (1-adamantyl) acetyl] -4-isopropyl-l, 3-oxazolan-2-one (29). 1-Adamantane-acetic acid (1.0 g, 5.14 mmol) was dissolved in CH2C12 (15 m) containing 1 drop of DMF. The mixture was stirred magnetically at 5 ° C under a nitrogen atmosphere and oxalyl chloride (1.1 equiv., 0.72 g, 496 μL, 5.66 mmol) was added dropwise over 20 min. After 2 h, the dichloromethane was evaporated in vacuo. The residual oil was dissolved in benzene (10 L) and concentrated to provide compound 28 (1.09 g, 100%) as a pale yellow oil which was used as such in the next reaction. To a solution of (45) - (-) - 4-isopropyl-2-oxazolidinone (0.66 g, 5.14 mmol) in anhydrous THF (10 mL) at -40 ° C was added n-BuLi dropwise (3.22 mL, 1.6 M in hexanes, 5.14 mmol). After 30 min at -40 ° C, the reaction mixture was cooled to -78 ° C. The crude acid chloride 28 (1.09 g, 5.14 mmol) dissolved in THF (1 mL) was added dropwise. The mixture was then magnetically stirred at 0 ° C for 1 h. Ethyl acetate was added and the organic phase was washed with 20% aqueous citric acid, saturated aqueous NaHCO3, dried (MgSO4), filtered and concentrated. The residual solid was purified by flash chromatography on silica gel, eluting with hexane / ethyl acetate (5/1) to give pure 29 (1.26 g, 80%) as a white solid: mp 105-22 107 ° C; [a] D + 62 ° (c 1.54, MeOH); H-NMR (400 MHz, CDC13) d 4.48-4.44 (m, ÍH), 4.24-4.16 (m, 2H), 2.91 (d, J = 14 Hz, 1H), 2.71 (d, J = 14 Hz, HH), 2.38-2.33 (m, HH), 2.00-1.94 (m, 3H), 1.75-1.61 (m, 12H), 0. 92 (d, J = 7.5 Hz, 3H), 0.89 (d, J = 7.5 Hz, 3H); 13 C-NMR (100.6 MHz, CDC13) d 171.21, 154.08, 62.84, 58. 54, 46.71, 42.23, 36.73, 33.81, 28.63, 28.53, 18. 04, 14.68; FAB MS m / z: 306 (MH +); HRMS cale for C? 8H28N03 (MH +) 306.2069, found: 306.2058; Anal (Ci8H27N03) C, H, N.
Acid (2S) -2- (1-adamantyl) -2-azidoethaneiso (31). The oxazolidinone 29 (4.2 g, 13.7 mmol) was dissolved in THF (15 L) and added dropwise over 15 min to a solution of potassium bis (trimethylsilyl) amide (20.1 mL, 0.69 M in THF, 13.9 mmol) a -78 ü C. After 45 min at -78 ° C, 2,4,6-triisopropylbenzenesulfonyl azide (4.9 g, 15.8 mmol) in THF (10 L) at -78 ° C was added in one portion to the enolate. After 5 min, glacial acetic acid (4.6 equiv, 3.8 g, 3.61 mL, 63.2 mmdl) was added and the mixture was stirred at 40 ° C for 1 h. The tetrahydrofuran was evaporated under reduced pressure and the residue was dissolved in a mixture of EtOAc and water. The organic phase was washed with saturated aqueous NaHCO3, followed by brine, dried (MgSO4), filtered and concentrated. The residual oil was dissolved in 75 mL of hexane / ethyl acetate (2/1). After 16 h at 25 ° C, the white precipitate was removed by filtration and the filtrate was concentrated to give an oily residue which was filtered through a pad of silica, washed with hexane / ethyl acetate (8/1). This material (30 crude, 1.3 g, 27%) was dissolved in THF / water (70 mL, 3/1) and H202 (4 equiv, 30%, 1.69 mL, 15 mmol) was added at 0 ° C followed by Li0H -H20 (2.1 equiv, 0.33 g, 7.88 mmol). After 45 min, 10% aqueous Na2S203 (48 mL) and solid NaHCO3 (0.22 g) were added. The reaction mixture was concentrated and ca 50 mL of water was added. The aqueous phase was washed with chloroform (4 times), acidified at 0 ° C with 15% HCl, and extracted with EtOAc (3 times). The combined EtOAc extracts were dried (MgSO 4), filtered and concentrated to give compound 31 (0.79 g, 90% crude) as a white solid: mp 110-112 ° C; [a] D -36 ° (c 1.60, MeOH); iH-NMR (400 MHz, CDC13) d 3.60 (s, ÍH), 2.16-2.00 (, 3H), 1.76-1.62 (m, 12H); 13 C-NMR (100.6 MHz, CDC13) d 174.19, 72.51, 38.68, 37.50, 36.51, 28.19; FAB MS m / z: 236 (MH +); HRMS cale for C 12 H 18 N 302 (MH +) 236.1399, found: 236.1389; Anal (C? 2H17Ñ302) C, H, N. 4- [(AS) -4-isopropyl-2-oxo-l, 3-oxazolan-3-xl] -2,2-dimethyl-4-oxobutanoate benzyl (34). n-Butyl lithium (2.4 mL, 1.6 M in hexanes, 3.9 mmol) was added dropwise to a solution of (45) - (-) -4-isopropyl-2-oxazolidinone (0.5 g, 3.9 mmol) in THF ( 5 mL) at -40 ° C under a nitrogen atmosphere. After 30 min at -40 ° C, the reaction mixture was cooled to -78 ° C and 2, 2-dimethylsuccinic anhydride (0.5 g, 3.9 mmol) dissolved in THF (2 mL) was added dropwise. The mixture was then magnetically stirred at 0 ° C for 1 h. Ethyl acetate was added and the organic phase was washed with 20% aqueous citric acid, brine, dried (MgSO 4), and concentrated to give crude as a pale yellow solid (1.0 g) which was dissolved in acetonitrile (5 g). mL) at 0 ° C. 1,8-Diazabicyclo [5.4.0] undec-7-ene (0.59 g, 583 μL, 3.9 mmol) and benzyl bromide (0.67 g, 463 μL, 3.9 mmol) were added and the reaction mixture was then stirred at 25 ° C for 16 h. The acetonitrile is evaporated to the brine. The residue was partitioned between EtOAc and 20% aqueous citric acid. The organic phase was washed with water, brine, dried (MgSO 4), filtered and concentrated. The residue was purified by flash chromatography on silica gel eluting with hexane / ethyl acetate (4/1) to give pure 34 (0.88 g, 61% of 32) 22 as a colorless oil.; [a] D + 56 ° (c 1.30, CHC13); 1 H-NMR (400 MHz, CDCl 3) d 7.31-7.28 (m, 5H), 5.11 (m, 2H), 4.36-4.32 (m, 1H), 4.23-4.15 (m, 2H), 3.25 (s, 2H) , 2.30-2.22 (m, ÍH), 1.33 (s, 3H), 1.31 (s, 3H), 0. 86 (d, J = 1 Hz, 3H), 0.81 (d, J = 7 Hz, 3H); 13 C-NMR (100.6 MHz, CDC13) d 176.73, 170.75, 154.03, 136.25, 128.35, 127.88, 66.24, 63.44, 58.27, 45.31, 40.24, 28.31, 25.62, 25.58, 17.84, 14.56; FAB MS m / z: 348 (MH +); HRMS for C? 9H26N05 (MH +) 348.1811, found: 348.1822; Anal (C? 9H25N05) C, H, N.
Benzyl 3-azido-2,2-dimethylsuccinic acid (36). The oxazolidinone 34 (8.67 g, 24.9 mmol) was dissolved in THF (27 mL) and added dropwise over 15 min to a solution of potassium bis (trimethylsilyl) amide (36.5 L, 0.69 M in THF, 25.2 mmol) at -78 ° C. After 45 min at -78 ° C, 2,4,6-triisopropylbenzenesulfonyl azide (8.89 g, 28.7 mmol) in THF (15 mL) at -78 ° C was added in one portion to the enolate. After 5 min, glacial acetic acid (4.6 equiv, 6.90 g, 6.56 mL, 0.12 mol) was added and the mixture was stirred at 35-40 ° C for 90 min. The tetrahydrofuran was evaporated under reduced pressure and the residue was dissolved in a mixture of EtOAc and water. The organic phase was washed with saturated aqueous NaHCO3, brine, dried (MgSO4), filtered and concentrated.
The residue was dissolved in 150 mL of hexane / ethyl acetate (2/1). After 16 h at 25 ° C, the white precipitate was removed by filtration and the filtrate was concentrated to give an oil which was filtered through a pad of silica and rinsed with hexane / ethyl acetate (5/1). This pale yellow oil (crude 35, 5.37 g, 55%) was dissolved in THF / water (260 mL, 3/1) and H202 (4 equiv, 30%, 6.24 mL, 55 mmol) was added at 0 ° C, followed by LiOH-H20 (2.1 equiv, 1.22 g, 29 mmol). After 45 min, 10% aqueous Na2S203 (175 mL) and solid NaHCO3 (0.81 g) were added. The tetrahydrofuran was evaporated and the aqueous phase was extracted with chloroform (continuous liquid-liquid extraction, 24 h). The aqueous phase was then acidified with concentrated HCl at 0 ° C, and extracted with EtOAc (3 times). The combined EtOAc extracts were dried (MgSO 4), filtered and concentrated. The residual oil was purified by flash chromatography on Merck silica gel eluting with ethyl acetate / acetic acid (400/1) to give compound 36 (0.82 g, 21%) as a colorless oil; [to; D -74 ° (c 1.43, CHC13); 3-H-NMR (400 MHz, CDC13) d 7.39 -7.32 (m, 5H), 5.20-5.12 (m, 2H), 4.48 (s, ÍH), 1.34 (s, 3H), 1.29 (s, 3H); 13 C-NMR (100.6 MHz, CDCl 3) d 174.74, 173.86, 135.42, 128.56, 128.35, 128.13, 67.86, 67.18, 45.98, 23.01, 20.29; FAB MS m / z: 278 (MH +); HRMS for C? 3H? 6N304 (MH +) 278.1141, found: 278.1130; Anal (C? 3H15N30) C, H, N.
Example 6. NI- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- ((SS) -2-methyl-1 - [((SS) -2-methyl) -l- [(methylcarboxamido) methyl] carboxamidopropyl) carboxamido] propylcarboxamido) butanediamide (37, Table 1) . This compound was prepared in solid phase using the active ketone resin (Example 1). The final purification was carried out by preparative HPLC.
HPLC (system A) 97%, (system B) 98%; IR (KBr)? 3400-3000 (br) 1637 154 cm 1H H - NMR (400 MHz, DMS0-d6), 1: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.51 and 8.49 (d, J = 6.0 and 6.0 Hz, 0.5H), 8.08-8.02 (m, 2H ), 7.83-7.76 (m, 2H), 7.47-7.30 (m, 1.5H), 6.95-6.88 (, 2H), 4.61-4.52 (m, 1.5H), 4.26-4.06 (m, 2.5H), 3.79 -3.67 (m, 2H), 2.67-2.32 (m, 2H), 1.99-1.93 (m, 2H), 1.85 (s, 3H), 1.26 and 1.25 (d, J = 4.1 and 3.8 Hz, 1.5H), 1.06 (d, 6.7 Hz, 1.5H), 0.84-0.79 (m, 12H); FAB MS m / z: 553 (MH +); HRMS cale for C22H36F3N607 (MH +) 553.2597, found: 553.2617.
Example 7. NI- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2S) -6-amino-2- ((SS) -1- [((SS) -1- [( ÍS) -2-hydroxy-1- (methylcarboxamido) ethyl] carboxamido-2- (4-hydroxy enyl) ethyl) carboxamido] -2-methylpropylcarboxamido) hexanamide (38, Table 1). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 98%, (system B) 99%; IR (KBr)? 3500-2800 (br), 1643, 1516 cm-1; -H-NMR (400 MHz, DMS0-d6), 3: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomer in Pi, d 9.14 (s, ÍH), 8.71 and 8.68 (d, J = 5.7 and 5.7 Hz, 0.25H), 8.08 and 8.03 (d, J = 8.0 and 8.0 Hz, ÍH), 7.91 (t, J = 7.1 Hz, 2H), 7.75 (quartet, J = 7.9 Hz, ÍH), 7.63 ( br s, 3H), 7.57 and 7.56 (d, J = 10.9 and 8.9 Hz, 1H), 7.02-6.95 (m, 2.75H), 6.61 (d, < J = 8.3 Hz, 2H), 4.94 (m, 1 H), 4.71-4.62 (m, 0.25H), 4.43 (m, 1H), 4.26 (m, 2H), 4.13 (m, 1.75H), 3.48 (t, J = 5.7 Hz, 2H), 2.92 ( m, 1H), 2.74 (br, 3H), 1.95 (q, J = 6.8 Hz, ÍH), 1.83 (s, 3H), 1.62 (m, ÍH), 1.51 (m, 2H), 1.08 (m, 3H ), 1.08 and 1.07 (d, J = 6.7 and 6.6 Hz, 3H), 0.86-0.81 (m, 6H); FAB MS m / z 661 (MH +); HRMS cale for C29H44F3N608 (MH +) 661.3173, found: 661.3195.
Example 8. NI- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (2/3) -2-t ((IS) -2-methyl-1 - [(1S) -2- methyl-l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (39, Table 1). This compound was prepared in solution using standard coupling methods from 3 (Example 3) and the oxidation of trifluoromethyl alcohol with the Moffatt-Pfitzner method. The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system B) 100%; IR (KBr)? 3600-2800, 1636, 1546 cm_1; iH-RM (400 MHz, DMSO-dβ), 3: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Plf d 8.53 (m, 0.25H), 8.09 and 8.06 (d, J = 7.9 and 7.4 Hz, ÍH); 7.89 and 7.87 (d, J = 4.9 and 5.4 Hz, ÍH), 7.72 and 7.70 (d, J = 4.5 and 4.0 Hz, 1H), 7.46 (d, J = 9.8 Hz, 0.4H), 7.37-7.30 (m , 1.5H), 6.96-6.89 (m, 2.6H), 4.54 (m, 1H), 4.21-4.06 (m, 3H), 2.48-2.34 (m, 2H), 2.00-1.91 (m, 2H) , 1871 and 1868 (s, 3H), 1.27 and 1.25 (d, J = 4.5 and 3.9 Hz, 0.75H), 1.06 (d, 6.9 Hz, 2.25H), 0.84-0.80 (m, 12H); FAB MS m / z: 496 (MH +); HRMS cale for C 20 H 33 F 3 N 5 O 6 (MH +) 496.2382, found: 496.2387.
Example 9. NI- (3,3, 3-trifluo o-1-me il-2-oxopropyl) - (2S) -2-. { [(SS) -2-methyl-l- [(methylcarboxamido) -propyl] carboxamido} butanediamide (40, Table 1). This compound was prepared by solution using standard coupling methods. The final oxidation of ethyl trifluoro alcohol was carried out with the Moffatt-Pfitzner method. HPLC (system A) 100%, (system D) 98%; IR (KBr)? 1685, 1655, 1627 ern-1; iH-RM (400 MHz, DMS0-d6), 7: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pl d 8.19 and 8.11 (2 xd, J = 7.6 and 7.6 Hz, ÍH), 7.94 (m, ÍH), 7.38 (m, 2H), 6.91 (m, 3H), 4.51 (m, 1H), 4.10 (m, 2H), 2.40 (, ÍH), 1.94 (m, ÍH), 1.89 and 1.87 (2 xs, 3H), 1.26 (m, 0.4H), 1.07 (d, J = 5.4 Hz, 2.6H), 0.83 (t, J = 6.3 Hz, 6H); FAB MS m / z: 397 (MH +), 415 (M + 19); HRMS cale for C? 5H24F3N40s (MH +) 397.1699, found: 397.1712.
Example 10 NI- (3, 3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2- (methylcarboxamido) butanediamide (41, Table 1). This compound was prepared in solution using standard coupling methods from 3 (Example 3) and the oxidation of trifluoromethyl alcohol with the Moffatt-Pfitzner method. HPLC (system A) 99%, (system D) 99%; IR (KBr)? 3387, 1696, 1653 cm ~ l; 1 H NMR (400 MHz, DMSO-dβ) 1: 1 mixture of diastereoisomers in Pi, d 8.65 and 8.59 (2 xd, J = 6.1 and 5.7 Hz, 0.1H), 8.09 and 8.04 (2 xd, J = 7.9 and 7.9 Hz, 1H), 7.53 and 7.36 (2 xd, J = 9.2 and 9.2 Hz, ÍH), 7.31 (br s, ÍH), 6.96-6.87 (, 3H), 4.61-4.47 (m, ÍH), 4.05-4.15 (m, 1H), 2.49 and 2.31 (m, 2H), 1.25 (dd, J = 7.0 and 4.4 Hz, 0.3H), 1.07 (dd, J = 7.0 and 3.5 Hz, 2.7H); FAB MS m / z: 298 (MH +), 316 (M + 19); HRMS for C? OH? 5F3N3? 4 (MH +) 298.1015, found: 298.1026.
Example 11. NI- (3,3,3-trifluoro- (IR) -methyl-2-oxopropyl) - (2 S) -2- [(SS) -2-methyl-1- (methylcarboxamido) propyl] carboxamidobutanediamide (42, Table 1). This compound was prepared in solution using standard coupling methods from 3 (Example 3) and the oxidation of trifluoromethyl alcohol with the Moffatt-Pfitzner method. The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system D) 97%; IR (KBr)? 1685, 1671, 1638 crn "1; 1 E NMR (400 MHz, DMSO-de), 20: 1 hydrated / non-hydrated mixture, d 8.11 (d, J - 7.5 Hz, ÍH), 7.95 (d, J = 8.0 Hz, 1H), 7.35 (br s, 1H), 7.32 (d, J = 9.3 Hz, ÍH), 6.92 (br s, ÍH), 4.52 (q, J = 7.2 Hz, ÍH), 4.11 (m , 2H), 2.45 (m, 2H), 1.95 (m, ÍH), 1.26 (d, J = 6.9 Hz, 0.05H), 1.07 (d, J = 6.9 Hz, 2.95H), 0.84 (t, J = 6.6 Hz, 6H); FAB MS m / z: 397.3 (MH +), 415.3 (M + 19); HRMS cale for C? 5H23F3N403 (MH +) 397.1699, found: 397.1707.
Example 12. NI- (3, 3, 3-trifluoro- (1S) -methyl-2-oxopropyl) - (2S) -2-. { [(1S) -2-methyl-1- (methylcarboxamido) propyl] carboxamido} butanediamide (43, Table 1). This compound was separated from 42 by preparative HPLC. HPLC (system A) 97%, (system D) 100%; IR (KBr)? 1685, 1663, 1626 cm "3-; HH-RM (400 MHz, DMSO-d6) hydrated form only, d 8.19 (d, J = 7.5 Hz, HH), 7.93 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 9.6 Hz, ÍH), 7.34 (br s, ÍH), 6.90 (br s, ÍH), 4.51 (q, J = 6.9 Hz, 1H), 4.11 (m, 2H), 2.45 (m , 2H), 1.93 (, ÍH), 1.06 (d, J = 6.9 Hz, 3H), 0.84 (m, 6H), FAB MS m / z: 397 (MH +), 415 (M + 19); C? 5H23F3N403 (MH +) 397.1699, found: 397.1712.
Example 13. NI- (1-ethyl-3, 3, 3-trifluoro-2-oxopropyl) - (2S) -2- [((SS) -2-methyl-1- [(SS) -2-methyl- 1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido} butanediamide (44, Table 1). This compound was prepared in solid phase using the substituted activated ketone resin as the ethyl analog (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D) 100%; IR (KBr)? 1640 crn-1; 1 H-NMR (400 MHz, DMS0-d 6), 7: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pl 7 d 8.55-8.47 (m, 0.14H), 8.09 and 8.05 (2 xd, J = 7.3 and 7.3 Hz, ÍH), 7.90-7.87 (m, 1H), 7.71-7.68 (m, ÍH), 7.40-7.24 (m, 2H), 6.92-6.77 (m, 3H), 4.59-4.52 (m , ÍH), 4.23-4.15 (, 2H), 3.96-3.87 (m, 1H), 2.67-2.32 (m, 2H), 2.01-1.90 (m, 2H), 1.87 and 1.86 (2 xs, 3H), 1.79 -1.71 (m, ÍH), 1.40-1.28 (m, ÍH), 0.89-0.74 (m, 15H); FAB MS m / z: 510 (MH +), 528 (M + 19); HRMS cale for C2? H35F3N506 (MH +) 510.2539, found: 510.2558.
Example 14. NI- (1- (3,3, 3-trifluoro-l-propyl-2-oxopropyl) - (2S) -2- [((SS) -2-methyl-l- [(1S) - 2-methyl-1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (45, Table 1) This compound was prepared by the same procedure as for 3 (Example 3), except that 1-nitroethane was replaced by 1-nitrobutane The coupling conditions of the standard solution were used to prepare the peptide inhibitor with the final oxidation of the trifluoromethyl alcohol using the Moffatt-Pfitzner method.The purification was carried out by preparative HPLC, HPLC (system A) 89%, (system D) 99%: IR (KBr)? 3280, 1663, 1637, 1546 cm-1; 1 H NMR (400 MHz, DMSO-d6), 7: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.50 (m, 0.25H), 8.18 (d, J = 7.3 Hz, 0.12H), 8.07 (m, ÍH), 8.02 (d, J = 7.95 Hz, 0.25H), 7.89 and 7.88 (2 xd , J = 8.6 and 8.9 Hz, HH), 7.77 (d, J = 8.3 Hz, 0.12H), 7.70 (d, J = 8.6 Hz, HH), 7.46-7.24 (m, 2H), 6.97-6.72 (m , 2.7H), 4.63-4.51 (m, 1H), 4.30 and 4.28 (2 xd, J = 6.7 and 6.7 Hz, 0.12H), 4.24-4.13 (m, 2H), 4.09-3.96 (m, ÍH), 2.50 -2.32 (m, 2H), 2.02-1.91 (m, 2H), 1.87 (s, 3H), 1.72-1.57 (, ÍH), 1.43-1.21 (m, 2H), 1.18-1.04 (id, ÍH), 0.89-0.75 (m, 15H); FAB MS m / z: 524 (MH +), 542 (M + 19); HRMS cale for C22H37F3N506 (MH +) 524.2696, found: 524.2705.
Example 15. NI- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2 S) -2-E ((SS) -2-methyl-l- [(SS) -2-methyl) -l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] pentanediamide (46, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 86%, (system D) 82%; IR (KBr)? 3281, 3079, 1647 cm "1; 1 H NMR (400MHz, DMS0-d6), 2: 3 hydrated / non-hydrated mixture, 1.2: 1 mixture of diastereoisomers in Plf d 8.72 (d, J = 5.7 Hz, 0.16H ), 8.70 (d, J = 5.7 Hz, 0.18H), 7.90-8.02 (m, 0.8H), 7.90 (d, J = 8.9 Hz, ÍH), 7.68-7.74 (m, ÍH), 7.56-7.61 ( m, 0.3H), 7.10-7.21 (rn, ÍH), 6.90-7.00 (m, ÍH), 6.70-6.75 (m, ÍH), 4.60-4.69 (m, 0.4H), 4.07-4.29 (m, 3.7 H), 2.00-2.09 (m, 2H), 1.90-1.98 (m, 2H), 1.86 (s, 3H), 1.83-1.61 (m, 2H), 1.28 (d, J = 7.0 Hz, 0.5H), 1.27 (d, J = 7.0 Hz, 0.6H), 1.08 (d, J = 6.5 Hz, 1.4H), 1.07 (d, J = 6.4 Hz, 1.2H), 0.81- 0.84 (m, 12H); FAB MS m / z: 510 (MH +); HRMS cale for C2? H35F3N506 (MH +) 510. 2539, found: 510.2521.
Example 16 (3S) -3- [((SS) -2-methyl-l- [(1S) -2-methyl-1- (methylcarboxamido) rovyl] carboxamidopropyl) carboxamido] -3- (3, 3, 3-trifluoro) acid -l-methyl-2-oxopropyl) carbamoyl propanoic (47, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 96%, (system B) 97%; IR (KBr)? 3500-2800 (br), 1639, 1546 cm "1; 1 E-NMR (400 MHz, DMS0-d6), hydrated form only, 1: 1 mixture of diastereomers in Pi, d 12.29 (br s, ÍH), 8.22 and 8.14 (d, J = 7.8 and 7.9 Hz, ÍH), 7.89 (m, ÍH), 7.70 (m, ÍH), 7.40 and 7.35 (d, J = 8.8 and 9.4 Hz, ÍH), 6.93 (t, J = 7.8 Hz, 2H), 4.53 (m, ÍH), 4.17 (m, 2H), 4.11 (q, J = 6.9 Hz, 1H), 2.67-2.58 (m, 1H), 2.47 (m, ÍH), 1.94 (m, 2H), 1.87 (s, 3H), 1.06 (m, 3H), 0.83 (m, 12H); FAB MS m / z: 497 (MH +); HRMS cale for C20H32F3N4O7 (MH +) 497.2223; found: 497.2237.
Example 17. NI- [(SS) -1- ((1S) -2-hydroxy-l- [(3,3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] tyl-carbamoyl) -2 -methylpropyl] - (2 S) -3-methyl-2- (methylcarboxamido) butanamide (48, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 94%, (system B) 99%; IR (KBr)? 3500-2800, 1637, 1543 cm "1; ^ -H-NMR (400 MHz, DMSO-de), hydrated form only, 1: 1 mixture of diastereomers in Pi, d 7.94 (d, J = 7.9 Hz, 1H) , 7.90 (d, J = 8.9 Hz, ÍH), 7.73 and 7.71 (d, J = 5.4 and 4.9 Hz, ÍH), 7.55 and 7.50 (8.9 and 9.4 Hz, ÍH), 6.93 (br m, 2H), 4.31 -4.10 (m, 4H), 3.57-3.47 (m, 3H), 1.97 (m, 2H), 1.87 (s, 3H), 1.09 and 1.08 (d, J = 6.9 and 6.9 Hz, 3H), 0.86-0.81 (m, 12H); FAB MS m / z: 469 (MH +); HRMS cale for C? 9H32F3N406 (MH +) 469.2274, found: 469.2261.
Example 18. NI- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2 S) -6-amino-2- [((1 S) -2-methyl-1- [(IS ) -2-methyl-1- (methylcarboxamido) rovyl] carboxamidopropyl) carboxamido] hexanamide (49, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 98%, (system B) 96%; IR (KBr)? 3227, 1638, 1545, 1189 crn "1; 1 E NMR (400 MHz, DMSO-d6), hydrated form only, 1: 1 mixture of diastereomers in Px, d 7.99 (2 xd, J = 7.9 and 7.4 Hz, 1H ), 7.88 (d, J = 8.8 Hz, ÍH), 7.71 and 7.70 (2 xd, J = 8.7 and 8.4 Hz, ÍH), 7.58 (t, J = 8.9 Hz, 1H), 6.99 (m, 2H), 4.29-4.09 (m, 4H), 2.73 (m, 2H), 2.00-1.92 (m, 2H), 1.93 (s, 3H), 1.63-1.46 (m, 4H), 1.28-1.25 (m, 2H), 1.09 and 1.07 (2 xd, J = 6.9 and 6.9 Hz, 3H), 0.85-0.82 (m, 12H); FAB MS (FAB) m / z: 510 (MH +), 528 (M + 19); HRMS cale for C22H39F3N5Os (MH +) 510.2903, found: 510.2888.
Example 19. NI- [(1 S) -2-methyl-l- ((1S) -2- (1,3-thiazol-4-yl) -1 - [(3,3, 3-tri luoro-l -methyl-2-oxopropyl) -carbamoyl] ethylcarbamoyl) propyl] - (2 S) -3-methyl-2- (methylcarboxamido) utanamide (50, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 98%, (system D) 98%; IR (KBr)? 3276, 3084, 1638 was "!; iH-RM (400MHz, DMSO-d6), 1: 1.2 hydrated / non-hydrated, 1.2: 1 mixture of diastereomers in Pr, d 8.99 (d, J = 5.4 Hz, 0.55H ), 8.98 (d, J = 5.4 Hz, 0.45H), 8.77 (d, J = 5.7 Hz, 0.19H), 8.71 (d, J = 5.7 Hz, 0.23H), 8.09-8.17 (, 1H), 7.84 -7.88 (m, 1H), 7.67-7.90 (m, 1.43H), 7.55 (d, J = 8.9 Hz, 0.25H), 7.29-7.34 (, ÍH), 6.94 (br s, 0.75H), 4.58- 4.73 (m, 1.5H), 4.06-4.18 (m, 2.5H), 2.97-3.17 (m, 2H), 1.86-1.93 (, 2H), 1.86 (s, 1.65H), 1.85 (s, 1.35H) , 1.22 (d, J = 6.7 Hz, 0.61H), 1.21 (d, J = 6.7 Hz, 0.74H), 1.07 (d, J = 7.0 Hz, 0.93H), 0.98 (d, J = 6.7 Hz, 0.72 H), 0.76-0.80 (m, 12 H); FAB MS m / z: 536 (MH +), 554 (M + 19); HRMS cale for C22H33F3N505S (MH +) 536.2154, found: 536.2170.
Example 20. N 4, N 4 -dimethyl-N 1 - (3, 3, 3-trifluoro-l-methyl-2-oxopropyl) - (2 S) -2- [((SS) -2-methyl-1 - [( 1S) -2-methyl-l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (51, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system B) 99%; IR (KBr) 1638 cm "1; ifi-RM (400 MHz, DMS0-d6), 1: 1 mixture of diastereomers in Px, d 8.56-8.49 (m, 0.1H), 8.10-8.03 (m, 0.8H) , 7.89- 7.86 (, 0.8H), 7.73-7.70 (m, 0.8H), 7.44-7.41 (m, ÍH), 6.95-6.80 (m, 1.5H), 4.63-4.54 (m, ÍH), 4.20- 4.06 (m, 3H), 2.94-2.93 (m, 3H), 2.80-2.78 (m, 3H), 2.67-2.62 (m, 2H), 1.99-1.93 (m, 2H), 1.87-1.86 (m, 3H) ), 1.30-1.25 (m, 0.5H), 1.07-1.06 (m, 2.5H), 0.84- 0.83 (m, 12H); FAB MS m / z: 524 (MH +); HRMS cale for C 22 H 37 F 3 N 506 (MH +) 524.2696, found: 524.2710.
Example 21. NI- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -4-methyl-2- [((1S) -2-methyl-1 - [(1S) - 2-methyl-1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] pentanamide (52, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 97%, (system D) 97%; IR (KBr)? 3268, 3080, 1632 cm "1; ifi-NMR (400MHz, DMS0-d6), 1: 1.2 hydrated / non-hydrated, 1.2: 1 mixture of diastereomers in Pi, d 8.72 (d, J = 6.5 Hz, 0.25H ), 8.70 (d, J = 6.5 Hz, 0.3H), 7.91-8.10 (m, ÍH), 7.84-7.88 (, ÍH), 7.74-7.80 (m, ÍH), 7.55 (d, J = 8.6 Hz, 0.2H), 7.53 (d, J = 8.9 Hz, 0.25H), 6.90-6.96 (m, ÍH), 4.57-4.68 (, 0.5H), 4.26-4.39 (m, ÍH), 4.15-4.21 (m, ÍH), 4.05-4.14 (m, 1.4H), 1.87-1.95 (m, 2H), 1.85 (s, 3H), 1.53-1.63 (m, ÍH), 1.30-1.50 (m, 2H), 1.26 (d , J = 7.0 Hz, 0.8H), 1.25 (d, J = 7.0 Hz, ÍH), 1.07 (d, J = 6.7Hz, 0.8H), 1.06 (d, J = 6.7 Hz, 0.6H), 0.78- 0.88 (m, 18H); FAB MS m / z: 495 (MH +); HRMS cale for C22H38F3 405 (MH +) 495.2794, found: 495.2803; Anal (C22H37F3N405-H20) C, H, N.
Example 22. NI- [(SS) -2-methyl-l- ((1S) -2-phenyl-1 - [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) ropil] - (2S) -3-methyl-2- (methylcarboxamido) utan-amide (53, Table 2). This compound was prepared by solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 98%, (system B) 100%; IR (KBr)? 3280, 1636, 1546 ern "1; Í-H-NMR (400 MHz, DMSO-d6) 1: 1 mixture of diastereomers in Pi, d 8.03 (m, ÍH), 7.86 (m, 1H), 7.64 (m, ÍH), 7.22 (, 5H), 7.19 (m, ÍH), 6.93 and 6.90 (2 xd, J = 14.3 and 13.7 Hz, 2H), 4.63-4.54 (m, 2H), 4.16-4.05 (m, 4H) , 3.00-2.66 (m, 2H), 1.86 (s, 3H), 1.85 (, 2H), 1.25, 1.21, 1.09 and 0.96 (4 xd, J = 7.4, 6.8, 6.9 and 7.9 Hz, 3H), 0.79 ( m, 12H); FAB MS m / z: 529 (MH +), 547 (M + 19); HRMS cale for C 25 H 36 F 3 N 4? 5 (MH +) 529.2638, found: 529.2619.
Example 23. Nl- [(SS) -2-methyl-l- ((1S) -2-methyl-l- [(3,3,3-trifluoro-l-methyl-2-oxopropyl) aarbamoyl] -propylcarbamoyl) ropil] - (2S) -3-methyl-2- (methylcarboxamido) butanamide (54, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). HPLC (system A) 84%, (system B) 83%; IR (KBr) 1633 cm "1; ifi-NMR (400 MHz, DMSO-de) 1: 1 mixture of diastereomers in Pi, d 8.78 (d, J = 5.5 Hz, 0.25H), 8.71 (d, J = 5.5 Hz, 0.25H), 7.88-7.60 (m, 3.5H), 6.98-6.87 (m, 1H), 4.70-4.64 (m, 0.5H), 4.22-4.09 (m, 3.5H), 1.97-1.91 (m , 3H), 1.86 (s, 3H), 1.28-1.26 (m, 1.7H), 1.09-1.07 (m, 1.3H), 0.88-0.78 (m, 18H); FAB MS m / z: 481 (MH +); HRMS for C2? H36F3N405 (MH +) 481.2638; found: 481.2627.
Example 24. NI- [(1S) -2-methyl-1 - ((1S) -1- [(3, 3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] ethyl-carbamoyl) propyl] - (2S) -3-methyl-2- (methylcarboxamido) butanamide (55, Table 2). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system B) 99%; IR (KBr)? 3264, 1627, 1552 cm "1; ^ -H-NMR (400 MHz, DMS0-d6) 1: 1 mixture of diastereomers in Px, d 7.98 and 7.92 (2 xd, J = 7.3 and 7.3 Hz, ÍH), 7.88 -7.83 (m, 1H), 7.72 (d, J = 8.8 Hz, ÍH), 7.58 (t, J = 9.4 Hz, ÍH), 6.95 and 6.91 (2 x br s, 2H), 4.33-4.06 (, 4H ), 1.95 (m, 2H), 1.87 (s, 3H), 1.17-1.13 (m, 3H), 1.08 (d, J = 6.9 Hz, 3H), 0.85-0.81 (m, 6H); FAB MS m / z: 453 (MH +), 471 (M + 19); HRMS cale for C? 9H32F3N405 (MH +) 453.2325, found: 453.2338.
Example 25. NI- [(1S) -2-methyl-l- ((IR) -1- [(3,3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] ethyl-carbamoyl) propyl] - (2S) -3-methyl-2- (methylcarboxamido) butanamide (56, Table 2). This compound was separated from 55 by preparative HPLC. HPLC (system A) 99%, (system B) 99%; IR (KBr) 1634 cm "1; HI-RM (400 MHz, DMS0-d6) 1: 1 mixture of diastereomers in Px, d 8.63 (d, J = 5.5 Hz, 0.1 H), 8.56 (d, J-4.5 Hz, 0.1H), 8.18-8.12 (m, 0.2H), 8.01 (d, J = 1 Hz, 0.3H), 7.96 (d, J = 7.5 Hz, 0.3H), 7.89-7.74 (, 1.8H) , 7.65 (d, J = 9 Hz, 0.3H), 7.60 (d, J = 9.5 Hz, 0.3H), 6.96 -6.92 (m, 1.3H), 4.68-4.59 (m, 0.3H), 4.34-4.25 (m, ÍH), 4.19-4.02 (m, 2.7H), 1.98-1.89 (m, 2H), 1.86 (s, 3H), 1.31-1.29 (m, 0.8H), 1.19-1.07 (m, 5.2H) ), 0.86-0.82 (, 12H); FAB MS m / z: 453 (MH +); HRMS for C? 9H32F3 4? 5 (MH +) 453.2325; found: 453.2338.
Example 26. N4, N4-dimethyl-NI- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2- [((1S) -1- [(SS) -2- methyl-l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (57, Table 3). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D, pH 7.4) 100%; GO (KBr)? 3283, 1642 cm "1; ifi-NMR (400 MHz, DMSO-d6), 19: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.05 and 8.01 (2 xd, J = 7.6 and 7.6 Hz, 1H), 7.91-7.82 (m, 2H), 7.43 and 7.39 (2 xd, J = 9.2 and 9.2 Hz, 1H), 7.01-6.80 (m, 2H), 4.61-4.52 (m, ÍH), 4.22-4.00 (m, 3H), 2.94 and 2.93 (2 xs, 3H), 2.80 (s, 3H), 2.71-2.59 (, 2H), 2.00-1.90 (m, ÍH), 1.87 and 1.86 (2 xs, 3H), 1.73-1.61 (m, 1H), 1.57-1.45 (m, 1H), 1.27 and 1.26 (2 xd, J = 6.7 and 6.7 Hz, 0.15H), 1.06 (d, J = 6.0 Hz, 3H), 0.87-0.78 (m, 9H); FAB MS m / z: 510.3 (MH +), 528.3 (M + 19); HRMS cale for C2? H35F3Ns? 6 (MH +) 510.2539, found: 510.2526.
Example 27. N 4, N 4 -dimethyl-N 1 - (3, 3, 3-trifluoro-l-methyl-2-oxopropyl) - (2 S) -2- [((1S) -2, 2-dimethyl-l- [(1 S) -2-methyl-1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] -butanediamide (58, Table 3). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system B) 99%; IR (KBr)? 3500-2900, 1640, 1538 c "1; ^ - E-NMR (400 MHz, DMSO-de), hydrated form only, 1: 1 mixture of diastereomers in Pi, d 8.20 and 8.13 (d, J = 7.4 and 7.2 Hz, ÍH), 7.91 and 7.91 (d, J = 9.0 and 8.7 Hz, ÍH), 7.56 and 7.55 (d, J = 9.3 and 9.3 Hz, ÍH), 7.50 and 7.43 (d, J = 9.3 and 9.3 Hz, 1H), 6.93 (br s, 1H), 6.81 (br s, ÍH), 4.57 (m, ÍH), 4.23 (m, 2H), 4.11 (m, ÍH), 2.95 and 2.94 (s, 3H), 2.80 and 2.80 (s, 3H ), 2.72-2.57 (m, 2 H), 1.95 (m, J = 6.9 Hz, 1H), 1.87 (s, 3H), 1.07 and 1.06 (d, J = 8.1 and 6.6 Hz, 3H), 0.89 (s) , 9H), 0.83 (d, J = 6.8 Hz, 6H); FAB MS m / z: 538 (MH +); HRMS for C 23 H 39 F 3 N 5 6 6 (MH +) 538.2852, found: 538.2843.
Example 28. N4, N4-dimethyl-Nl- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- [((SS) -3,3-dimethyl-1- [ (SS) -2-methyl-1- (methylcarboxamido) propyl] carboxamidobutyl) carboxamido] butanediamide (59, Table 3). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D, pH 7.4) 100%; IR (KBr)? 3285, 1644 cm "1; 1 E NMR (400 MHz, DMSO-de), 16: 1 mixture of hydrated / non-hydrated, 3: 2 mixture of diastereomers in Pi, d 8.05 (d, J = 8.3 Hz, ÍH) , 7.91-7.79 (m, 2H), 7.43 and 7.39 (2 xd, J = 9.2 and 9.2, ÍH), 7.11-6.66 (br s, 2H hydrate), 4.62-4.49 (m, 1H), 4.36-4.25 ( m, ÍH), 4.16-4.02 (m, 2H), 2.94 and 2.93 (2 xs, 3H), 2.80 (s, 3H), 2.71-2.55 (m, 2H), 1.99-1.88 (m, ÍH), 1.85 and 1.84 (2 xs, 3H), 1.67-1.58 (m, 1H), 1.46 (dd, J = 14.2 and 8.9 Hz, ÍH), 1.28-1.25 (m, 0.2H), 1.06 (d, J = 6.7 Hz , 2.8H), 0.86 (s, 9H), 0.83 (d, J = 6.7 Hz, 3H), 0.81 (d, J = 6.7 Hz, 3H); FAB MS m / z: 552 (MH +), 570 (M + 19); HRMS cale for C24H4iF3N506 (MH +) 552.3009, found: 552.3031.
Example 29. N4, N4-dimethyl-Nl- (3,3,3-tri-loro-1-methyl-2-oxopropyl) - (2S) -2- [((S) -1- (1-adamantyl) - 1- [(SS) -2-methyl-1- (methylcarboxamido) propyl] carboxamido methyl) carboxamido] butan-diamide (60, Table 3). This compound was prepared in solution using standard coupling methods and oxidized with Dess-Martin periodinnan. The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D) 99%; IR (KBr)? 3293, 1641, 1533 cm "1; ifi-NMR (400 MHz, DMS0-d6), 4: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.58 (d, J = . 5 Hz, 0.1H), 8.47 (d, J = 6.5 Hz, 0.1H), 8.22-8.12 (m, ÍH), 7.94-7.91 (m, 1H), 7.49-7.38 (m, 1.9H), 6.94 ( br s, 1H), 6.86 (br s, ÍH), 4.60-4.52 (m, 1H), 4.24- 4.20 (m, ÍH), 4.13-3.98 (m, 2H), 2.94 (s, 1.5H), 2.93 (s, 1.5H), 2.79 (s, 3H), 2.74-2.57 (m, 2H), 2.00-1.94 (m, ÍH), 1.94-1.86 (m, 3H), 1.87 (s, 3H), 1.64-1.48 (m, 12H), 1.26-1.25 (m, 0.6H), 1.07 (d, J = 6.5 Hz, 2. 4H), 0.82 (d, J = 6.5 Hz, 6H); FAB MS m / z: 616 (MH +); HRMS cal e for CagH? S FaNsOe (MH +) 61 6. 3322, found: 61 6. 3335; Ana l (C29H44 F3N5? 6-H20) C, H, N Example 30. (3S) -3 - ((lS) -2- (dimethylcarbamoyl) -l- [(3,3,3-tri-loro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) -2 acid, 2-dimethyl-3- [(SS) -2-methyl-1- (methylcarboxamido) rovyl] carboxamidopropanoic (61, Table 3). This compound was prepared in solution using standard coupling methods. The residue of β, β-dimethyl aspartic acid was incorporated as the β-benzyl ester derivative. The oxidation of the trifluoromethyl alcohol was carried out with the Dess-Martin periodinane. The final purification was carried out by preparative HPLC. HPLC (system A) 96%, (system D) 98%, IR (KBr)? 1654, 1532 cm "1; 1-H-NMR (400 MHz, DMS0-d6) 1: 1 mixture of diastereomers in Pi, d 7.98-7.89 (m, 2 H), 7.76 (d, J = 7.5 Hz, 0.5H), 7.67 (d, J = 7.5 Hz, 0.5H), 7.53 (d, J = 9 Hz, 0.5H), 7.48 (d, J = 9 Hz, 0.5H), 6.92 (br s, 1H), 6.83 (br s, 1H) , 4.75 (d, J = 9.5 Hz, ÍH), 4.57-4.51 (, ÍH), 4.27-4.19 (m, ÍH), 4.18-4.03 (m, 1H), 2.94 (s, 1.5H), 2.93 (s) , 1.5H), 2.80 (s, 3H), 2.71-2.57 (m, 2H), 2.04-1.95 (m, ÍH), 1.87 (s, 3H), 1.08-1.05 (m, 9H), 0.85 (d, J = 6.5 Hz, 3H), 0.83 (d, J = 6.5 Hz, 3H); FAB MS m / z: 568 (MH +); HRMS cale for C23H37F3N508 (MH +) 568.2594; found: 568.2505.
Example 31. N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2 S) -2 - [(1S) -2, 2-dimethyl-l- ( methylcarboxamido) propyl 3 carboxamidobutanediamide (62, Table 4). This compound was prepared in solution using standard coupling methods and the trifluoromethyl alcohol was oxidized with the Dess-Martin periodinnan. The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system E) 100%; IR (KBr)? 1640 cm "1; iH-NMR (400 MHz, DMSO-de), 1: 1 mixture of diastereomers in Pi, d 8.06 and 7.94 (2 xd, J = 7.6 and 7.3 Hz, ÍH), 7.69 (d, J = 8.9 Hz, 1H), 7.22 and 7.21 (2 xd, J = 8.9 and 9.2 Hz, 1H), 4.42-4.34 (m, ÍH), 4.01 and 3.96 (2 xd, J = 8.9 and 8.9 Hz, ÍH), 3.95 -3.89 (m, ÍH), 2.77 (d, J = 3.2 Hz, 3H), 2.62 (s, 3H), 2.59-2.42 (m, 2H), 1.72 (d, J = 3.8 Hz, 3H), 1.09 and 0.89 (2 xd, J = 7.0 and 6.7 Hz, 3H), 0.73 and 0.72 (2 xs, 9H); FAB MS m / z: 439 (MH +), 457 (M + 19); HRMS for C? 8H3oF3 4? 5 (MH +) 439.2168, found: 439.2154.
Example 32. N4, N4-dimethyl-Nl- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- ((1S) -1- [(4-hydroxyphenyl-ethyl) carboxamido] -2, 2-dimethylpropylcarboxamido) butanediamide (63, Table 4). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out on a preparative HPLC. HPLC (system A) 97%, (system B) 95%; IR (KBr)? 1636 cm "1; iH-NMR (400 MHz, DMSO-d6) 16: 1 mixture of hydrated / non-hydrated, 1.4: 1 mixture of diastereomers in Px, d 9.20-9.00 (m, ÍH), 8.23 and 8. 13 (2 x d, J = 7.4 and 7.4 Hz, ÍH), 7.79 (2 x d, J = 9. 0 and 9.0 Hz, ÍH), 7.41 (2 x d, J = 9.0 and 9.0 Hz, 1H), 6.99 (d, J = 8.6 Hz, 2H), 6.99-6.75 (m, 1.5H hydrate), 6.65-6.62 (m, 2H), 4.62-4.52 (m, ÍH), 4.20 and 4.17 (2 xd , J = 9.0 and 9.0 Hz, ÍH), 4.15-4.05 (m, ÍH), 2.96 and 2.95 (2 x s, 3H), 2.80 (s, 3H), 2.72- 2.60 (m, 4H), 2.50-2.45 (m, ÍH), 2.45-2.35 (m, 1H), 1. 27 (d, J = 7.0 Hz, 0.1H), 1.75 (m, 2.8H), 0.86 (s, 9H); FAB MS m / z: 545 (MH +), 563 (M + 19); HRMS cale for C 25 H 36 F 3 N 4 6 (MH +) 545.2587, found 545.2602.
Example 33. N 4, N 4 -dimethyl-NI- (3, 3, 3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- [(1S) -1- (isobutylcarboxamido) -2,2- dimethylpropyl] carboxamidobutanediamide (64, Table 4).
This compound was prepared in solution using standard coupling methods. The final purification was carried out by preparative HPLC. HPLC (system A) 95%, (system B) 99%; IR (KBr)? 1636 cm "1; ifi-NMR (400 MHz, DMSO-de), 19: 1 mixture of hydrated / non-hydrated, 1: 1 mixture of diastereomers in Px, d 8.21 and 8.11 (2 xd, J = 7.2 and 7.2 Hz, ÍH), 7.73 and 7.72 (2 xd, J = 9.0 and 9.0 Hz, ÍH), 7.44 and 7.40 (2 xd, J = 9.6 and 9.3 Hz, 1H), 7.1-6.7 (br, 1.7H hydrate), 4.60-4.52 (m, ÍH), 4.21 and 4.19 (2 xd, J = 9.0 and 8.7 Hz, ÍH), 4.15-4.04 (, ÍH), 2.96 and 2.95 (2 xs, 3H), 2.80 (s, 3H), 2.74-2.58 (m, 2H), 2.14 and 2.12 (2 xd, J = 13.2 and 12.9 Hz, ÍH), 2.08 (m, ÍH), 1.96 (m, ÍH), 1.07-1.04 ( m, 3H) 0.90 (s, 9H), 0.87 (d, J = 6.6 Hz, 3H), 0.86 (d, J = 6.6 Hz, 3H); FAB MS m / z: 481 (MH +), 499 (M + 19); HRMS for C2? H36F3N4? 5 (MH +) 481.2638, found 481.2627.
Example 34. N4, N4-dimethyl-Nl- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2 - [(l3) -2, 2-dimethyl-1- (neopentylcarboxamido ) ropil] carboxamidobutanediamide (65, Table 4). This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system B) 99%; IR (KBr)? 1635 crn "1; ifi-RM (400 MHz, DMSO-de), 29: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.20 and 8.10 (2 xd, < J = 7.3 and 7.0 Hz, ÍH), 7.62 (d, J = 8.9 Hz, ÍH), 7.44 and 7.43 (2 xd, J = 9.2 and 9.2 Hz, ÍH), 6.95-6.75 (m, 1.7H), 4.56 (quintet, J = 6.7 Hz, ÍH), 4.19 and 4.16 (2 xd, J = 8.9 and 8.9 Hz, 1H), 4.12-4.05 (m, ÍH), 2.95 and 2.94 (2 xs, 3H), 2.79 (s) , 3H), 2.75-2.60 (m, 2H), 2.20-2.15 (m, HH), 2.04 and 2.01 (2 x d, J = 12.7 and 12.4 Hz, HH), 1.07-1.04 (m, 3H), 0.95 (s, 9H), 0.90 (s, 9H); FAB MS m / z: 495 (MH +), 513 (M + 19); HRMS cale for C22H38F3N405 (MH +) 495.2794, found 495.2777.
Example 35. N 4, N 4 -dimethyl-NI- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- ((1S) -1- [(3, 3-dimethyl- butyl) amino] -2,2-dimethylpropylcarboxamido) butanediamide (66, Table 4) . This compound was prepared by solid phase using the activated ketone resin (Example 1). The final reductive amination in the terminal amine of t-butyl glycine (0.3 mmol) was carried out in solid phase by the addition of 3,3-dimethylbutyraldehyde (376 mL, 3.0 mmol) in DMF (15 mL with acetic acid (150 mL). , Y NaBH3CN (63 mg, 1 mmol) for 20 h. After removal of the solvent, the resin was cut in the usual manner. After purification by preparative HPLC the compound was obtained as a white solid (20.6 mg) after lyophilization. HPLC (system A) 99%, (system D) 97%; IR (KBr)? 2960, 1667 cm "1; OH-RM (400 MHz, DMSO-d6), 1: 1 mixture of diastereomers in Plf d 8.79-8.72 (, 2H), 8.04 (br s, 1H), 7.78 (d, J = 9.0 Hz, 0.5H), 7.67 (d, J = 9.0 Hz, 0.5H), 6.97-6.96 (m, 1.3H), 6.91 (s, 0.6H), 4.82-4.75 (, 1H), 4.15-4.11 ( m, 1H), 3.61 (d, J = 9.5 Hz, 1H), 2.94 (m, 3H), 2.87 (m, ÍH), 2.79 (, 3H), 2.70-2.65 (m, 3H), 1.66-1.57 ( m, ÍH), 1.51-1.44 (m, ÍH), 1.29-1.24 (m, ÍH), 1.08 (d, J = 7.0 Hz, 3H), 1.02 (s, 5.4H), 0.99 (s, 3.6H) 0.89 (s, 3.6H), 0.89 (s, 5.4H), FAB MS m / z: 481 (MH +), 499 (M + 19); HRMS cale for C22H4oF3N4? 4 (MH +) 481.3002, found: 481.2991.
Example 36. 4N, 4N-Dimethyl-1 N- (3,3,3-trifluoro-l-methyl-2-oxopropyl-2- [1- (er-butoxy-aryl-amino) -2,2-dimethyl- (IS) -propylcarboxamido- (2S) -butanediamide (67, Table 4) This compound was prepared in solid phase using the activated ketone resin (Example 1) The final purification was performed by preparative HPLC (system A) 99%, (system B) 98%; IR (KBr)? 3500-2900, 1641, 1510 cm "1; XH-NMR (400 MHz, DMSO-d6), 5: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Pi, d 8.56 (br d, J = 5.1 Hz, 0.15H), 8.10-8.00 (m, ÍH), 7.50-7.46 (m, 1H), 6.93-6.82 (m, 1.7H), 6.50-6.49 (, 1H), 4.62 (m, ÍH) , 4.12 (m, ÍH), 3.84 (m, ÍH), 2.95 (m, 3H), 2.80 (s, 3H), 2.65 (m, 2H), 1.38 (s, 9H), 1.26 (d, J = 6.6 Hz, 0.45H), 1.06 (d, J = 6.6 Hz, 2.55H), 0.88 (s, 9H), FAB MS m / z: 497 (MH +), HRMS cale for C2iH36F3N406 (MH +) 497.2587, found: 497.2601.
Example 37. N4, N4-Dimethyl-NI- (3,3, 3-trifluoro-1-methyl-2-oxopropyl-2- [1- (tert-butylaminocarbonyl-amino) -2,2-dimethyl- (IS) -propylcarboxamido3 - (2S) -butandiamide (68, Table 4). This compound was prepared in solution using standard coupling methods and the oxidation of trifluoromethyl alcohol with the Dess-Martin periodinane. The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system D) 100%; IR (KBr)? 1641 cm "1; ^ -H-NMR (400 MHz, DMSO-de), 1: 1 mixture of diastereomers in Pi, d 8.15 and 8.09 (2 xd, J = 7.3 and 7.0 Hz, HI), 7.51 and 7.43 ( 2 xd, J = 8.9 and 9.2 Hz, ÍH), 6.00 (s, ÍH), 5.97-5.93 (, ÍH), 4.58-4.53 (m, ÍH), 4.11-4.03 (m, ÍH), 3.94-3.91 ( m, 1H), 2.95 (d, J = 5 Hz, 3H), 2.79 (s, 3H), 2.67-2.60 (m, 2H), 1.20 (s, 9H), 1.07-1.05 (m, 3H), 0.86 (s, 9H); FAB MS m / z: 496 (MH +), 514 (M + 19); HRMS cale for C21H37F3N505 (MH +) 496.2747, found: 496.2765.
Example 38. N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-l-methyl-2-oxopropyl) - (2/3) -2- [((1 S) -1- [(dimethyl- amino) methyl] carboxamido-2,2-dimethylpropyl) carboxamido] butanediamide (69, Table 4). This compound was prepared in solution using standard coupling methods and the oxidation of trifluoromethyl alcohol with the Dess-Martin periodinane. The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D) 98%; IR (KBr)? 1654, 1540, 1186 ern "1; XH-NMR (400 MHz, DMS0-d6), 9: 1 hydrated / non-hydrated mixture, 1: 1 mixture of diastereomers in Px, d 9.61 (br s, 1H), 8.68 and 8.66 (2 xd, J = 9.0 and 8.5 Hz, 1 H), 8.37 and 8.30 (2 xd, J = 7.5 and 7.0 Hz, ÍH), 7.48 and 7.45 (2 xd, J = 9.0 and 9.0 Hz, 1H), 6.95-6.87 (m, 2H ), 4.63-4.54 (m, 1H), 4.35-4.32 (m, 1H), 4.12-3.94 (m, 3H), 2.95-2.94 (m, 3H), 2.95-2.94 (m, 3H), 2.80-2.78 (m, 9H), 2.72-2.56 (m, 2H), 1.26-1.24 (m, 0.3H), 1.07-1.06 (m, 2.7H), 0.92 (s, 9H); FAB MS m / z: 482 (MH +), 500.1 (M + 19); HRMS cale for C2oH35F3N505 (MH +) 482.2590, found: 482.2599.
Example 39. 4- ((S) -1- ((1S) -2- (dimethylcarbamoyl) -l- [(3,3,3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] ethylcarbamoyl) -2 acid , 2-dimethylpropyl] carbamoylbutanoic acid (70, Table 4). This compound was prepared in solution using standard coupling methods. The final residue P was introduced by the opening of the glutaric anhydride in Et3N. The final purification was carried out by preparative HPLC. HPLC (system A) 100%, (system D) "100%; IR (KBr)? 1638, 1537, 1176 c" 1; ^ -H-NMR (400 MHz, DMS0-d6) 1: 1 mixture of diastereomers in Pi, d 8.22 and 8.12 (2 xd, J = 7.5 and 7.0 Hz, ÍH), 7.81-7.72 (m, 1H), 7.43 and 7.39 (2 xd, J = 9.0 and 9.0 Hz, ÍH), 6.93 (br s, 0.8H), 6.81 (br s, 0.8H), 4.60-4.52 (m, 1H), 4.21-4.18 (m, ÍH ), 4.12-4.05 (m, 1H), 2.95-2.94 (m, 3H), 2.79 (s, 3H), 2.68-2.63 (m, 2H), 2.28-2.14 (m, 4H), 1.73-1.66 (m , 2H), 1.11-1.05 (m, 3H), 0.89 (s, 9H); FAB MS m / z: 511.2 (MH +), 529 (M + 19); HRMS cale for C2? H34 3N407 (MH +) 511.2379, found: 511.2363; Anal (C2? H33F3N407-2H20) C, H, N.
Example 40. N 4, N 4 -dimethyl-NI- (3, 3, 3-trifluoro-1-methy1-2-oxopropyl) - (2_3) -2 - [(1 S) -l-amino-2, 2-dimethylpropi] carboxamidobutanediamide. This compound was prepared in solid phase using the activated ketone resin (Example 1). The final purification was carried out by preparative HPLC. HPLC (system D) 99%, (system E) 99%; IR (KBr)? 1670 c "1; ifi-NMR (400 MHz, DMS0-d6), 2: 1 hydrated / non-hydrated, 1: 1 mixture of diastereomers in Pi, d 8.87 and 8.80 (2 xd, J = 5.7 and 6.4 Hz, 0.4H), 8.56-8.49 (m, 1H), 8.02 (brs, 3H), 7.69 and 7.64 (2 xd, J = 9.0 and 9.2 Hz, 0.6H), 6.96, 6.95, 6.88 and 6.83 (4 xs, 1.3H), 4.73-4.63 (m, 1.4H), 4.14-4.08 (, 0.6H), 3.55-3.48 (m, 1H), 2.96 (s, 3H), 2.82T (s, 3H), 2.74-2.58 (m, 2H), 1.25 (d, J = 6.7 Hz, ÍH), 1.08 and 1.07 (2 xd, J = 6.7 and 6.7 Hz, 2H), 1.01-0.96 (m, 9H); FAB MS m / z: 397 (MH +), 415 (M + 19); HRMS cale for Ci6H28F3N404 (MH +) 397.2063, found 397.2077.
Example 41. N4, N -dimethyl-NI- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (2S) -2- [(SS) -1-hydroxy-2, 2-dimethylpro? id] carboxamidobutanediamide. This compound was prepared in solution using standard coupling methods. The 2-hydroxy isobutyric acid radical was introduced as the acetyl derivative. Oxidation of the trifluoromethyl alcohol with the Dess-Martin periodinane was followed by cutting the acetate group with aq NaOH. The final purification was carried out by preparative HPLC. HPLC (system A) 92%, (system D) 99%; IR (KBr)? 1641 cm "1; H-NMR (400 MHz, DMS0-d6), 2: 1 hydrated, non-hydrated, 1: 1 mixture of diastereomers in Px, d 7.83 and 7.79 (2 xd, J = 7.9 and 7.6 Hz, ÍH), 7.73-7.67 (, 0.3H), 7.57 and 7.52 (2 xd, J = 9.2 and 9.2 Hz, ÍH), 7.04-6.82 (m, 2H, hydrated), 5.69-5.47 (m, 1H), 4.65 -4.55 (m, ÍH), 4.19-4.05 (m, ÍH), 2.95 and 2.94 (2 xs, 3H) 2.80 and 2.79 (2 xs, 3H), 2.79-2.53 (, 3H), 1.39-1.02 (m, 3H), 0.87 (s, 9H); FAB MS m / z: 398 (MH +), 416 (M + 19); HRMS cale. for C? 6H27F3N3? 5 (MH +) 398.1903, found: 398.1892.
Example 42. N4, N4-dimethyl-Nl- (3,3,3-trifluoro-l-methyl-2-oxopropyl) - (2S) -2- (neopentylcarboxamido) -butanediamide. This compound was prepared in solution using standard coupling methods and the final oxidation of the trifluoromethyl alcohol was carried out with the Dess-Martin periodinnan. The final purification was carried out by preparative HPLC. HPLC (system D) 97%, (system E) 99%; IR (KBr)? 1638 crn "1; 1-H-NMR (400 MHz, DMSO-d6), 2: 1 mixture of diastereomers in Px, d 7.80-7.77 (m, ÍH), 7.27 and 7.22 (2 xd, J = 9.2 and 9.2 Hz, ÍH), 4.49-4.42 (m, ÍH), 3.98-3.92 (m, ÍH), 2.80 (s, 3H), 2.64 (s, 3H), 2.55-2.40 (m, 2H), 1.84 (s, 2H), 1.10 and 0.92 (2 xd, J = 6.9 and 6.6 Hz, 3H), 0.79 (s, 9H), FAB MS m / z: 382 (MH +), 400 (M + 19); HRMS cale for C? 6H27F3N304 (MH +) 382.1954, found: 382.1968.
Example 43. N4, N4-dimethyl-NI- (3,3,4,4, 4-pentafluoro-l-methyl-2-oxobutyl) - (2¿3) -2- [(SS) -2, 2 - dimethyl-1- (neopentyl carboxamido) propyl] carboxamidobutanediamide (74, Table ) . Compound 11 (0.92 g, 3.14 mmol) was treated with 4 N HCl / dioxane (1.5 h) before it was concentrated in vacuo. The resulting hydrochloride salt (3.14 mmol) was combined with Boc-Asn (? -Me2) -OH (0.93 g, 3.45 mmol), TBTU (1.21 g, 3.77 mmol), HOBt (0.51 g, 1.2 mmol), and -Pr2NEt (1.64 mL, 9.42 mmol) in CH Cl2 (10 mL). After 3 h at rt, the mixture was extracted into EtOAc and washed with 1 N HCl, saturated aqueous NaHCO 3, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo. The residue was purified by flash chromatography (4: 1 EtOAc / hexane) to give the coupled product (0.594 g, 43%). HPLC (system A) 93%; ^ -H-NMR (CDCI3), 1.3: 1 mixture of diastereomers, d 6.92 (bs, 0.6H), 6.66 (d, J = 7.95 Hz, 0.4H), 5.80 (m, ÍH), 4.71 (bs, 0.4 H), 4.50 (m, 1.6H), 4.45-4.20 (m, 2H), 3.34 (dd, J = 16.85 and 16.85 Hz, 0.4H), 3.06-2.97 (m, 0.6H), 3.045 and 3.04 (2 xs, 3H), 3.02 and 3.00 (2 xs, 3H), 2.67 (dd, J = 7.63 and 7.63 Hz, 0.6H), 2.58 (dd, J = 16.85 and 16.85 Hz, 0.4H), 1.48 and 1.46 (2 xs, 9H), 1.32 (t, J = 7.95 Hz, 3H). The dipeptide (0.43 g, 0.99 mmol) was treated with 4 N HCl / dioxane (10 mL) for 2 h before it was concentrated in vacuo. The resulting hydrochloride salt (0.99 mmol) was combined with Boc-Tbg-OH (0.254 g, 1.1 mmol), BOP (0.487 g, 1.1 mmol), and i-Pr2NEt (0.52 mL, 3.0 mmol) in CH2Cl2 (10 mL). ). After 2.5 h at rt, the mixture was extracted into EtOAc and washed with 1 N HCl, saturated aqueous NaHC 3, and brine.
The organic phase was dried (MgSO 4), filtered and concentrated in vacuo. The residue was purified by flash chromatography (1: 1 EtOAc / hexane) to give the coupled product as a white solid (0.33 g, 60%). HPLC (system A) 99%; ^ -H-NMR (CDCI3), 1.3: 1 mixture of diastereomers, d 8.15 (m, 0.4 H), 7.73 (m, 0.6 H), 7.37 (m, 0.6 H), 7.10 (m, 0.4 H), 5.12 (m, 1 H), 4.77 (m, 0.7 H), 4.69 (m, 1.3 H), 4.25 (m, 2 H), 3.76 (, 1 H), 3.26 (dd, J = 15.9 and 15.9 Hz, 0.4 H), 3.16 (dd, J = 12.4 and 12.4 Hz, 0.6 H), 3.06 and 3.03 (2 xs, 3 H), 2.94 and 2.92 (2 xs, 3 H), 2.55 (dd, < J = 7.0 y 7.0 Hz, 0.6 H), 2.39 (dd, J = 11.4 and 11.4 Hz, 0.4 H), 1.46 and 1.45 (2 xs, 9 H), 1.37-1.25 (m, 3 H), 1.06 and 1.03 (2 xs, 9 H). This peptide (0.30 g, 0.67 mmol) was then treated with 4 N HCl / dioxane (10 mL) and concentrated in vacuo. The hydrochloride salt was combined with tert-bu lyl ether (0.094 μL, 0.74 mmol), BOP (0.33 g, 0.74 mmol), i-Pr2NEt (0.23 mL, 1. 34 mmol) in CH2Cl2 (10 mL) and stirred 3.5 h at rt.
The mixture was diluted with EtOAc and washed with 1 N HCl, saturated aqueous NaHCO 3, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo.
The residue was purified by flash chromatography to give the desired peptide as a white solid (0.297 g, 88%). HPLC (system A) 99%); iH-RM (CDCI3) d 7.79 (d, J = 7.6 Hz, 1 H), 7.32 (d, 8.3 Hz, 1 H), 6.18 (d, J = 6.7 Hz, 1 H), 5.21 (d, "= 6.7 Hz, ÍH), 4.82- 4.78 (m, ÍH), 4.39-4.31 (m, ÍH), 4.23-4.10 (m, 2H), 3.11-3.06 (dd, J = 16.2 and 15.7 Hz, ÍH), 2.99 (s, 3H), 2.91 ( s, 3H), 2.52 (dd, J = 16.2 and 15.9 Hz, ÍH), 2.18 (s, 2H), 1.27 (d, J = 6.7 Hz, 3H), 1.06 (s, 9H), 1.05 (s, 9H The alcohol (0.26 g, 0.48 mmol) thus obtained was combined with the Dess-Martin periodinnan (0.51 g, 1.19 mmol) in CH2Cl2 (5 mL) and stirred for 4 h.
The reaction mixture was diluted with EtOAc and treated with a 1: 1 mixture of 10% a2S2? 3: saturated NaHCO3 (10 mL) for 15 min. After extraction with EtOAc the organic phase was dried (MgSO 4), filtered and concentrated in vacuo. Pentafluoroethyl ketone was obtained by trituration with 3: 1 hexane / EtOAc to give 74 as a white solid (0.217 g, 84%). IR (KBr)? 1685, 1618 cm "1; ^ -H-NMR (DMSO-dg) 3: 1 mixture of diastereomers in P] _, d 8.20 and 8.14 (2 x d, J = 7.5 y 7. 5 Hz, HH), 7.66 (d, J = 9.0 Hz, HH), 7.51 (d, J = 8.8 Hz, HH), 6.91 (m, 2H), 4.58 (m, HH), 4.16 (m, 2H) , 2.95 and 2.80 (2 xs, 6H), 2.70 (m, 2H), 2.11 (m, 2H), 1.08 (d, J = 6.6 Hz, 3H), 0.95 (s, 9H), 0.91 (s, 9H); HRMS cale for C 23 H 38 F 5 N 4 O 5 (MH +) 545.2762, found: 545.2775.
Example 44. NI- [3- (benzylcarbamoyl) -3,3-difluoro-l-methyl-2-oxopropyl] -N 4, N 4 -dimethyl- (2/3) -2- [(SS) -2, 2- dimethyl-1- (neopentylcarboxamido) propyl] carboxamidobutanediamide (75, Table 5). Compound 13 (0.263 g, 0.73 mmol) was treated with 4N HCl / dioxane (6 mL) for 30 min before it was concentrated in vacuo. The resulting hydrochloride salt was combined with BOP (0.39 g, 0.88 mmol), Boc-Asn (? -NMe2) -OH (0.191 g, 0.73 mmol) and i-Pr2NEt (0.32 L, 1.83 mmol) in CH2Cl2 (5 L ). After 4 h, the reaction was poured into EtOAc and washed sequentially with 1N HCl, saturated aqueous aHC03, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo. The material was triturated with 3: 7 hexane: EtOAc to give a white solid (0.30 g, 82%). iH-RM (CDCI3) d 7.41-7.28 (m, 6H), 6.98-6.94 (m br, ÍH), 5.56-5.51 (m br, ÍH), 4.59-4.42 (m, 4H), 4.11-4.03 (m, 2H), 3.11-3.05 (, ÍH), 2.99 (s, 3H), 2.88 (s, 3H), 2.61-2.54 (m, 1H), 1.45 (s, 9H), 1.32 (d, J = 6.6 Hz, 3H). This material (0.27 g, 0.54 mmol) was treated with 4N HCl / dioxane (6 L) for 30 min before it was concentrated in vacuo. The hydrochloride salt (0.54 mmol) was combined with Boc-Tbg-OH (0.125 g, 0.54 mmol), BOP (0.286 g, 0.65 mmol) and i-Pr2NEt (0.23 mL, 1.35 mmol) in CH2C1 (3 mL) and it was stirred for 4 h. The mixture was diluted with EtOAc and washed sequentially with 1 N HCl, saturated aqueous NaHCO3, and brine before it was dried (MgSO4), filtered and concentrated in vacuo. The product was purified by flash chromatography using TLC grade silica gel to provide a white solid (0.25 g, 76%). ifí-NMR (CDC13) d 8.06 (s br, ÍH), 7.68 (s br, ÍH), 7.33-7.26 (m, 6H), 5.11 (d, J = 4.4 Hz, ÍH), 4.67 (dd, J = 14.6, 7.0 Hz, 1H), 4.50 (s br, ÍH), 4.32-4.22 (m, 3H), 4.13-4.07 (m, ÍH), 3.71 (d, J = 5.7 Hz, ÍH), 3.23-3.20 (m, 1H), 3.00 (s, 3H), 2.87 (s, 3H), 2.43-2.38 (dd, J = 15.9, 5.7 Hz, ÍH), 1.49 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H), 1.02 (s, 9H). This peptide (0.25 g, 0.41 mmol) was treated with 4 HCl N / dioxane (3 mL) and stirred 1 h before it was concentrated in vacuo. The hydrochloride salt (0.41 mmol) was combined with tert-bu lyl ether (52 μL, 0. 41 mmol), BOP (0.216 g, 0.49 mmol) and i-Pr2NEt (0.18 mL, 1.02 mmol) in CH2Cl2 (3 mL) and stirred 4 h. The mixture was diluted with EtOAc and washed sequentially with 1N HCl, saturated aqueous aHC 3, and brine before it was dried (MgSO 4), filtered and concentrated in vacuo. The product was purified by flash chromatography using TLC grade silica gel (5% MeOH / EtOAc) to give the fully worked-up peptide as a white solid (0.191 g, 77%). 1 H-NMR_ (CDCl 3) d 7.65 (d, J = 7.6 Hz, ÍH), 7.37-7.27 (m, 7H), 5.97 (d, J = 6.7 Hz, ÍH), 4.65-4.59 (m, 2H), 4.46 (d, J = 9.2 Hz, 1H), 4.35 (dd, J = 15.0, 5.4 Hz, ÍH), 4.28-4.24 (m, ÍH), 4.15-4.06 (m, 1H), 4.02 (d, J = 6.7 Hz, 1H), 3.16 (dd) , J = 15.9, 3.5 Hz, ÍH), 2.99 (s, 3H), 2.87 (s, 3H), 2.45 (dd, J = 15.6, 9.2 Hz, ÍH), 2.19 (dd, J = 18.1, 13.0 Hz, 2H), 1.24 (d, J = 7.0 Hz, 3H), 1.05 (s, 9H), 1.03 (s, 9H). The peptide (0.15 g, 0.245 mmol) was dissolved in CH2Cl2 (15 mL) and treated with Dess-Martin periodinnan (0.10 g, 0.245 mmol) and stirred at rt for 5 h. The mixture was diluted with EtOAc and treated with a 1: 1 mixture of 10% a 2 S 2 O 3: Saturated NaHC 3 (15 min). The organic phase was washed sequentially with saturated NaHC 3, 10% citric acid, and brine before it was dried (MgSO 4), filtered, and concentrated in vacuo. The final product was purified by preparative HPLC to give, after lyophilization, compound 75 as a white solid (0.115 g, 77%). IR (KBr)? 3293, 1680, 1635 cm "1, 1 E-NMR (CDCl 3), 1: 3 hydrated / non-hydrated mixture, d 9.65-9.55 (m, 0.25H), 8.99-8.47 (m, 0.75H), 8.20 -8.15 (m, 1H), 7.65-7.55 (m, ÍH), 7.40-7.20 (m, 6H), 6.55 (br s, 0.25H), 6.34 (br s, 0.75H), 4.80-4.72 (m, 0.25H), 4.59-4.53 (, ÍH), 4.42-4.26 (m, 2H), 4.26-4.16 (m, 1.5H), 4.11 (d, J = 8.3 Hz, 0.25), 2.96 (s, 2.25H) , 2.93 (s, 0.75H), 2.79 (s, 0.75H), 2.77 (s, 2.25H), 2.68 (m, 2H), 2.25-2.18 (m, ÍH), 2.05-1.95 (, ÍH), 1.24 (d, J = 7.0 Hz, 0.75H), 1.06 (d, J = 6. 6 Hz, 2.25H), 0.95 (s, 9H), 0.90 (s, 9H); 13c-NMR (100.6 MHz, DMSO-d6) d 197.32, 197.06, 196.79, 171.46, 171.08, 170.98, 170.49, 170.33, 169.86, 169.34, 161.15, 160.88, 160.62, 138.54, 138.04, 128.53, 128.40, 127.43, 127.27, 127.21, 126.98, 112.32, 109.68, 107.04, 60.45, 59.80, 50.04, 49.87, 49.58, 48.44, 48.26, 42.60, 42.36, 36.81, 35.03, 34.28, 33.89, 30.74, 29.84, 26.83, 15.54; HRMS cale for C3QH4gF2 5? G (MH +) 610.3416, found: 610.3395.
Example 45. NI- [2- (1, 3-benzoaeazol-2-yl) -1-methy1-2 -oxoethyl] -N4, N4-dimethyl- (23) -2-. { [(1/3) -2, 2-dimethyl-l- (eopentylcarboxamido) propyl] carboxamido} butanediamide (77, Table 5). A mixture of 16 (265 mg, 0.81 mmol) and 10% Pd in carbon (79 mg) in ethanol (20 mL) was stirred under a hydrogen atmosphere for 1 h. The solution was then filtered through a glass microfiber and concentrated under reduced pressure. The residue was dissolved in CH2Cl2 (6 mL) and Boc-Asn (? -NM? 2) -OH (222 mg, 0.85 mmol), HOBt (220 mg, 1.63 mmol), i-Pr2NEt (0.56 mL, 3. 25 mmol) and EDC (169 mg, 0.88 mmol). Additional--Pr2NEt was introduced to bring the pH above 8 and the stirring was continued overnight. The resulting mixture was diluted with EtOAc and washed sequentially with 10% citric acid, 10% C 2% and water and dried by passing through a glass wool plug. Flash chromatography (EtOAc) gave the desired compound (314 mg, 95%) .sup.-H-NMR (DMSO-dg) d 7. 82-7.66 (m, 3H), 7.42-7.33 (m, 3H), 6.78 and 6.73 (2 xd, J = 7.8 and 7.5 Hz, ÍH), 6.23-6.18 and 6.16-6.11 (2 xm, ÍH), 4.84 -4.82 and 4.66-4.50 (2 xm, ÍH), 4.33-4.17 (m, 2H), 2.88, 2.77, 2.74 (3 xs, 6H), 2.58-2.20 (m, 2H), 1.34 (s, 9H), 1.18-1.15 (m, 3H). This product was stirred in a mixture of CH2Cl2 (8 mL) and TFA (2 L) for 2 h. After removal of the solvent, the residual TFA was removed by azeotropic distillation with benzene using a rotary evaporator. The residue was dissolved in CH2Cl2 (6 mL) and Boc-Tbg-OH (188 mg, 0.81 mmol), HOBt, was added. (209 mg, 1.55 mmo 1), i-Pr NEt (0.54 mL, 3.10 mmol) and EDC (161 mg, 0.84 mmo 1). Additional i-Pr2 Et was introduced to bring the pH above 3 and the stirring was continued overnight. The resulting mixture was diluted with EtOAc and washed sequentially with 10% citric acid, Na 2 C 3 3 al % and water and dried by passing through a glass wool plug. Flash chromatography (EtOAc) gave the desired compound (264 mg, 62%). ÍH-RM (DMSO-dg) d 7.89-7.82 and 7.73-7.68 (m, 4H), 7.42-7.33 (m, 2H), 6.45 (d, J = 6.9 Hz, ÍH), 6.19 and 6.06 (2 xd, J = 6.0 and 5.4 Hz, 1H), 4.84 and 4.68 (2 xt, J = 5.1 and 6.3 Hz, ÍH), 4.61-4.51 (m, ÍH), 4.33-4.23 (m, ÍH), 3.84-3.77 (m, 1H), 2.89, 2.78, 2.72 (3 xs, 6H), 2.58-2.27 (m, 2H), 1.38 (s, 9H), 1.12 (m, 3H), 0.86 and 0.84 (2 xs, 9H) . This product was stirred in a mixture of CH2Cl2 (8 mL) and TFA (2 mL) for 2 h. After removal of the solvent, the residual TFA was removed by azeotropic distillation with benzene using a rotary evaporator. The residue was dissolved in CH2Cl2 (6 mL) and tert-butil to tertiary acid (64 mg, 0.50 mmol), HOBt (129 mg, 0.96 mmol), i-Pr2 Et (0.33 mL, 1.91 mmol) were added. and EDC (99 mg, 0.52 mmol).
Additional i-Pr2NEt was introduced to bring the pH above 8 and the stirring was continued overnight. The resulting mixture was diluted with EtOAc and washed sequentially with 10% citric acid, Na2C? 3 to 10%, water and dried by passing through a glass wool plug. Flash chromatography (EtOAc) gave the desired compound (162 mg, 62%) which was dissolved immediately in CH2Cl2 (8 mL). Dess-Martin periodinane (252 mg, 0.59 mmol) was added and the resulting mixture was stirred for 1 h. A 1: 1 mixture of 10% Na 2 S 2 → 3: saturated NaHCO 3 was introduced and stirring was continued until both layers were rinsed (10 min). The residue was extracted with CH 2 Cl 2, washed with aqueous NaHC 3, dried (MgSO 4), filtered and concentrated in vacuo. Flash chromatography using TLC grade silica gel (3% ethanol in EtOAc) provided the compound as a colorless oil. This material was dissolved in a minimum amount of CH 3 CN, diluted with water and lyophilized to provide the desired compound 77 as a white solid (99.3 mg, 61%). IR (KBr)? 3311, 1713, 1657 cm "1; HH-NMR (DMSO-dg), 2.7: 1 the mixture of diastereomers, d 8.28 (d, J = 5.7 Hz, HH), 8.16 and 8.08 (2 xd, J = 7.5 and 7.5 Hz, HH), 8.00 (d , J = 8.1 Hz, ÍH), 8.28 (d, J = 8.1 Hz, ÍH), 7.75-7.53 (m, 2H), 7.45-7.34 (m, ÍH), 6.99 and 6.75 (2 xs, ÍH, hydrate) , 5.30-5.22 and 4.41-4.35 (2 xm, 1H), 4.62 and 4.52 (2 xq, J = 6.0 and 7.2 Hz, ÍH), 4.18 and 4.13 (2 xd, J - 9.0 and 8.4 Hz, ÍH), 2.91 and 2.82 (2 xs, 3H), 2.77 and 2.71 (2 xs, 3H), 2.73-2.59 (m, 2H), 2.20 and 2.16 (2 xd, JAB = 12.6 and 12.9 Hz, ÍH), 2.03 and 2.02 (2 xd, JAB = 12.6 and 12.9 Hz, 1H), 1.42 and 1.06 (d, J = 7.2 and 6.9 Hz, 3H), 0.95-0.87 (m, 18H); HRMS cale for C28H42N5 ° 6 (MH +) 544.3135, found: 544.3154; Anal (C28H41N5O6) C, H, N.
Example 46. Diphenyl N 4, N 4 -dimethyl-N 1 - (1-aminoethylphosphinate) - (2/3) -2-. { [(SS) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (79, Table 5). To a hot solution of 1- (N-benzyloxycarbonyl) -aminoethylphosphonate (Oleksyszyn, J .; Subotkowska, L .; Mastalerz, P. Diphenil 1-aminoalkane? Hosphonates. Syn thesi s, 1979, 985-986) (8.50 g, 21.0 mmol) in ethanol (75 mL) was added to 4 N HCl / dioxane solution (5.25 mL, 21.0 mmol) and 10% Pd / C (850 mg. , 10% p / p). The mixture was vigorously stirred, leveled three times with hydrogen and stirred 16 h under a hydrogen atmosphere (balloon). The catalyst was filtered through Celite and the filtrate was concentrated in vacuo. The residual oil was crushed in Et2? (150 mL) until a white solid was obtained. This was filtered and dried to give 6.10 g (93%) of the corresponding hydrochloride salt. iH-NMR (DMSO-dg) d 9.18 (s, 3H), 7.40-7.44 (m, 4H), 7.24-7.27 (, 6H), 4.18 (dt, J = 7.2, 20.3 Hz, 1H), 1.61 (dd, Jx = 7.2 Hz, J2 = 18.0 Hz , 3H). A stirred solution containing Boc-Asn (? -NMe2) -OH (500 mg, 1.92 mmol), the hydrochloride salt of the above (663 mg, 2.11 mmol), I-Pr2NEt (836 μL, 4.80 mmol) and TBTU ( 677 mg, 2.11 mmol) in DMF (8 mL) was stirred initially at 0 ° C for 15 min, and then at rt for 3 h under a nitrogen atmosphere. The solution was poured into brine and the product was extracted with EtOAc (2 x 25 mL). The combined organic extracts were washed sequentially with 5% aqueous NaHCO 3, 1 M citric acid, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo to give 0. 975 g of an amorphous solid. The product was purified by flash chromatography (gradient 15-30% i-PrOH / hexane) to yield the coupled phosphonate derivative as an amorphous solid (0.81 g, 81%). HPLC (system A) 99.5%; ifi-NMR (CDCI3) 1: 1 mixture of diastereomers in Pi, d 7.65-7.35 (m, ÍH), 7.30-7.05 (m, 10H), 6.20-5.95 (, ÍH), 4.86-4.64 (m, ÍH), 4.54-4.42 (m, ÍH), 3.16-3.05 (, ÍH), 2.97-2.74 (m, 6H), 2. 56-2.41 (m, ÍH), 1.55-1.45 (m, 3H), 1.38 (s, 9H); FAB MS m / z: 520 (MH +), 420 (MH + -100). This material (0.75 g, 1.44 mmol) was treated with 4 N HCl / dioxane (30 min) before it was concentrated in vacuo. The hydrochloride salt (1.44 mmol) was combined with Boc-Tbg-OH (0.40 g, 1.73 mmol), TBTU (0.555 g, 1.73 mmol) and i-Pr2NEt (1.05 mL, 6.05 mmol) in DMF (8 mL) initially at 0 ° C (15 min) and then at 16 h. The reaction mixture was diluted with EtOAc and washed sequentially with 5% aqueous NaHC 3, 1 M citric acid, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo. Purification by flash chromatography using TLC grade silica gel (20% i -PrOH / hexane) gave the desired dipeptide fragment as a white solid (0.773 g, 85%). ^ -H-NMR (CDC13) d 8.06-7.80 (m, ÍH), 7. 50-7.40 (na, 1H), 7.38-7.10 (m, 10H), 5.24-5.12 (m, ÍH), 4.86-4.68 (m, 2H), 3.84-3.76 (m, 1H), 3.21-3.07 (m, ÍH), 2.95-2.78 (m, 6H), 2.58-2.35 (m, ÍH), 1.61- 1.49 (m, 3H), 1.43 (s, 9H), 0.96 (s, 9H); FAB MS m / z: 633 (MH +), 533 (MH + -100). This compound (0.70 g, 1.0 mmol) was treated with 4N HCl / dioxane (30 min) before it was concentrated in vacuo. The hydrochloride salt (1.0 mmol) was combined with tert-bu lyl ether (191 μL, 1.50 mmol), TBTU (0.385 g, 1.20 mmol) and i-P 2 NE (0.52 mL, 3.0 mmol) in DMF (10 mL) for 16 h. The reaction mixture was diluted with EtOAc and washed sequentially with 5% aqueous NaHC 3, 1 M citric acid, and brine. The organic phase was dried (MgSO 4), filtered and concentrated in vacuo. Purification was performed by preparative HPLC to give compound 79 (155 mg, 25%). IR (KBr)? 3289, 1642 c "1; ^ -H-NMR (DMSO-dg), 2: 1 mixture of diastereomers in Pi, d 8.35 (d, J = 8.9 Hz, 0.34H), 8.24 (d, J = 7. 3 Hz, 0.66H), 8.20 (d, J = 9.2 Hz, 0.66H), 8.15 (d, J = 7.6 Hz, 0.34H), 7.63 (d, J = 8.6 Hz, 0.66H), 7.58 (d, J = 8.6 Hz, 0.34H), 7.33-7.40 (m, 4H), 7.14-7.23 (m, 6H), 4.57-4.72 (m, 2H), 4.18 (d, J = 8.6 Hz, 0.66H), 4.17 (d, J = 8.6 Hz, 0.34H), 3.62 (s, wide, ÍH), 2.94 (s, ÍH), 2.88 (s, 2H), 2.79 (s, 1H), 2.77 (s, 2H), 2.59 -2.74 (m, 1H), 2.20 (d, J = 12.7 Hz, 0.66H), 2.17 (d, J = 12.7 Hz, 0.34H), 2.02 (d, J = 12.7 Hz, 0.66H), 1.98 (d , J = 12.7 Hz, 0.34H), 1.44 (d, J = 7.3 Hz, 1.5H), 1.39 (d, J = 7.3 Hz, 1.5H), 0.95 (s, 5.9H), 0.92 (s, 5.9H) ), 0.91 (s, 3.1H), 0.88 (s, 3.1H); HRMS cale for C32H48N4? 7P (MH +) 631.3260, found: 631.3279; Anal (C32H47N4O7 P) C, H, N.
Example 47. NI- [2- (1, 3-benzothiazol-2-yl) -1-methyl-2-oxo-ethyl] -N4, N4-dimethyl- (23) -2-. { [(1/3) -2, 2-di-methyl-l- (neopentylcarboxamido) propyl] carboxamido} butanediamide (80, Table 5). This compound was prepared from 14 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Moffatt-Pfitzner method. The final purification was carried out by preparative HPLC. HPLC (system A) 99%, (system D) 100%; IR (KBr)? 1642 cm "1; H-NMR (400 MHz, DMSO-de), the 1.5: 1 mixture of diastereomers, d 8.30-8.13 (m, 4H), 7.73-7.58 (m, 3H), 5.47-5.38 (m, 1H), 4.68-4.60 (, 1H), 4.18-4.10 (m, ÍH), 2.92 (s, 3H), 2.79 (s, 3H), 2.76-2.63 (m, 2H), 2.20 (d, J = 12.5 Hz, ÍH), 2.03 (d, J = 12.5 Hz, 1H), 1.43 and 1.37-1.28 (d, J = 7.3 Hz, m, 3H), 0.95 (s, 9H), 0.90 (s, 9H), FAB MS m / z: 560 (MH +); HRMS cale for C28H42 5? 5S (MH +) 560.2906, found: 560.2896.
Example 48. N 4, N 4 -dimethyl-N 1 - (1-methyl-2- [1, 33 oxazolo [4, 5-Ib] pyridin-2-yl-2-oxoethyl) - (2/3) -2-. { [(1/3) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (81, Table 5). This compound was prepared from 17 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Dess-Martin periodinane. The final purification was carried out by preparative HPLC. 1: 1 mixture of isomers; HPLC (system A) 97%, (system D) 98%; GO (KBr)? 1703, 1682, 1643 cm "1; iH-RM (400 MHz, DMSO-de) d 11.39 and 11.30 (2 xs, ÍH), 8.19 (d, J = 7.7 Hz, 1H), 7.96-7.83 (, ÍH) , 7.63-7.56 (m, 2H), 7.39-7.32 (m, 2H), 7.06-6.95 (m, ÍH), 4.65-4.46 (m, 2H), 4.25 and 4.22 (2 xd, J = 9.1 and 8.3 Hz , ÍH), 2.95 and 2.91 (2 xs, 3H), 2.77 and 2.66 (2 xs, 3H), 2.72-2.45 (m, 2H), 2.18-2.00 (m, 2H), 1.17 and 1.10 (2 xd, J = 6.8 and 6.8 Hz, 3H), 0.98-0.85 (, 18H), FAB MS m / z: 563 (M + 19), 585 (M + 18 + 23), HRMS cale for C27H43N607 (M + 19) 563.3193, found: 563.3207.
Example 49. N4, N4-dimethyl-Nl- [l-methyl-2- (6-methyl-l, 3-benzoxazol-2-yl) -2 -oxoethyl] - (2/3) -2-. { [(SS) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (82, Table 5). This compound was prepared from 20 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Dess-Martin periodinnan. The final purification was carried out by radial chromatography. HPLC (system A) 97%, (system C) 100%, (system D) 96%; IR (KBr)? 1713, 1650, 1642 cm "1; iH-NMR (400 MHz, DMS0-d6), 5: 1 mixture of diastereomers, d 8.36 and 8.15 (2 xd, J = 8.4 and 7.2 Hz, ÍH), 8.31 and 8.25 ( 2 xd, J = 5.1 and 5.7 Hz, HH), 7.89-7.86 (m, 1H), 7.73-7.69 (m, HH), 7.63-7.51 (m, HH), 7.41-7.36 (m, 1H), 5.29 -5.21 and 4.74-7.68 (2 xm, ÍH), 4.67-4.50 (m, ÍH), 4.14 and 3.91 (2 xd, J = 8.7 and 6.0 Hz, ÍH), 2.93-2.58 (m, 8H), 2.49 ( s, 3H), 2.23-2.18 (m, ÍH), 2.05-2.01 (m, ÍH), 1.45 and 1.41 (2 xd, J = 7.2 and 7.2 Hz, 3H), 0.97-0.87 (m, 18H); FAB MS m / z: 558 (MH +); HRMS cale for C29H44N506 (MH +) 558.3292, found: 558.3307; Anal (C28H43N506- * áH20) C, H, N. 50. N 4, N 4 -dimethyl-N 1 - [l-methyl-2- (5-methyl-l, 3-benzoxazol-2-yl) -2 -oxoethyl] - (2S) -2-. { [(1/3) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (83, Table 5). This compound was prepared from 19 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Dess-Martin periodinane. The final purification was performed by rapid chromatography. HPLC (system A) 96%, (system C) 99%, (system D) 94%; IR (KBr)? 1713, 1642 cm "1; ifi-NMR (400 MHz, DMSO-d6), 1.7: 1 mixture of diastereomers, d 8.38-8.06 (m, 2H), 7.81-7.67 (, 2H), 7.63-7.37 (m, 2H), 5.28-5.20 and 4.73-4.67 (2 xm, ÍH), 4.62 and 4.55-4.48 (q, J = 7.2 Hz, m, ÍH), 4.13 and 3.91 (2 xd, J = 8.4 and 6.3 Hz, 1H ), 2.93-2.46 (m, 8H), 2.49 (s, 3H), 2.21 and 2.20 (2 xd, JAB = 12.3 and 12.4 Hz, ÍH), 2.03 (2 xd, JAB = 12.3 Hz, ÍH), 1.45 and 1.41 (2 xd, J = 7.2 and 7.2 Hz, 3H), 0.97-0.87 (, 18H); FAB MS m / z: 558 (MH +); HRMS cale for C29H44N506 (MH +) 558.3292, found: 558.3307; Anal (C29H43N5Oe- ^ H20) C, H, N.
Example 51. N4, N4-dimethyl-Nl- [l-methyl-2- (4-methyl-l, 3-benzoxazol-2-yl) -2-oxoethyl] - (2/3) -2-. { [(1/3) -2, 2-dimethyl-l- (neopentylcarboxa ido) propyl] carboxamido} butanediamide (84, Table 5). This compound was prepared from 18 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Dess-Martin periodinnan. The final purification was performed by rapid chromatography. HPLC (system A) 97%, (system C) 99%, (system D) 94%; IR (KBr)? 1713, 1658, 1642 cm "1; 1H-NMR (400 MHz, DMSO-d6), 4: 1 mixture of diastereomers, d 8.38-8.08 (m, 2H), 7.71-7.15 (m, 4H), 5.32-5.24 and 4.74-4.68 (2 xm, ÍH), 4.63 and 4.55 (2 xq, J = 6.3 and 6.6 Hz, ÍH), 4.14 and 3.91 (2 xd, J = 8.4 and 6.6 Hz, 1H), 2.93-2.48 (m , 11H), 2.20 (d, AB = 12.6 Hz, 1H), 2.03 (d, JAB = 12.6 Hz, 1 H), 1.45 and 1.42 (2 xd, J = 7.2 and 7.2 Hz, 3H), 0.97-0.87 (m, 18H); FAB MS m / z: 558 (MH +); HRMS cale for C29H44N5? 6 (MH +) 558.3292, found: 558.3307; Anal (C29H43N506- ^ H20) C, H, N.
Example 52. N4, N4-dimethyl-NI- [l-methyl-2- (7-methyl-l, 3-benzoxazol-2-yl) -2-oxoethyl] - (2/3) -2-. { [(1S) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (85, Table 5). This compound was prepared from 21 using standard coupling methods and the oxidation of the heterocyclic alcohol with the Dess-Martin periodinnan. The final purification was performed by rapid chromatography. HPLC (system A) 97%, (system C) 97%, (system D) 92%; IR (KBr)? 1715, 1642 cm "1; ^ -H-NMR (400 MHz, DMSO-d6), 4: 1 mixture of diastereomers, d 8.37 and 8.15 (2 xd, J-8.1 and 7.5 Hz, IH), 8.34 and 8.28 ( 2 xd, J = 5.9 and 5.7 Hz, ÍH), 7.83-7.20 (m, 4H), 5.26 and 4.73-4.68 (quint, J = 6.3 Hz, m, ÍH), 4.62 and 4.57 (2 xq, J = 6.0 and 6.4 Hz, 1H), 4.14 and 3.90 (2 xd, J = 8.4 and 6.3 Hz, 1H), 2.93-2.58 (m, 8H), 2.54 (s, 3H), 2.21 and 2.20 (2 xd, JAB = 12.3 and 12.6 Hz, HH), 2.03 (d, JAB = 12.6 Hz, 1H), 1.45 and 1.42 (2 xd, J = 7.5 and 7.2 Hz, 3H), 0.97-0.87 (m, 18H); FAB MS m / z : 558 (MH +); HRMS cale for C29H4 N5? 6 (MH +) 558.3292, found: 558.3307; Anal (C29H43N5? 6- ^ H20) C, H, N.
Example 53. N4, N4-dimethyl-Nl- [l-methyl-2- (methyIcarbamoyl) -2-oxoethyl] - (2/3) -2-. { [(L / 3) -2,2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (86, Table 6). This compound was prepared according to the alternative procedure for the preparation of α-ketoamides (Example 2). The final purification was carried out by preparative HPLC.
HPLC (system C) 100%, (system D) 98%; IR (KBr)? 3320, 1645 cm "1; Í-H-NMR (400 MHz, DMSO-d6) d 8.61 and 8.56 (2 xm, ÍH), 8.18-8.05 (m, ÍH), 7.97 and 7.93 (2 xd, J = 6.4 and 6.3 Hz, ÍH), 7.60 (br d, J = 7.3 Hz, 1H), 5.03-4.90 (m, ÍH), 4.65-4.50 (m, ÍH), 4.13 and 4.12 (2 xd, J = 8.6 and 8.6 Hz, 1H), 2.94 (brs, 3H), 2.80 and 2.79 (2 xs, 3H), 2.71-2.55 (m, 5H), 2.20 (d, J = 12.4 Hz, 1H), 2.04 and 2.01 (2 xd , J = 12.7 and 12.7 Hz, ÍH), 1.23 and 1.22 (2 xd, J = 7.3 and 7.0 Hz, 3H), 0.95 (s, 9H), 0.91 (br s, 9H); FAB MS m / z: 484.3 (MH +), 506.3 (M + 23); HRMS cale for C23H 2N506 (MH +) 484.3135, found: 484.3148.
Example 54. NI- [2- (dimethylcarbamoyl) -l-methyl-2-oxoethyl] -N 4, N 4 -dimethyl- (2/3) -2-. { [(1/3) -2, 2-dimethyl-l- (neopentylcarboxamido) propyl] carboxamido} butandiamide. This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was carried out by preparative HPLC.
HPLC (system C) 99%, (system D) 99%; IR (KBr) 3302 , 1719, 1644 was "1; ^ -H-NMR (400 MHz, DMSO-d6) d 8.27 and 8.16 (2 xd, J = 6.4 and 6.7 Hz, 1H), 8.11 and 8.06 (2 xd, J = 7.6 and 7.9 Hz, HH), 7.59 and 7.58 (2 xd, J = 8.6 and 8.0 Hz, HH), 4.62-4.50 (m, 2H), 4.13 and 4.12 (2 xd, J = 8.0 and 8.6 Hz, HH), 2.94 (br s, 3H), 2.86 and 2.85 (2 xs, 6H), 2.80 (s, 3H), 2.72-2.56 (m, 2H), 2.23-2.16 (2 xd, J = 12.7 and 12.7 Hz, ÍH), 2.10-1.90 (m, ÍH), 1.30 and 1.29 (2 xd, J = 7.3 and 7.3 Hz, 3H), 0.95 (s, 9H), 0.90 (s, 9H); FAB MS m / z: 498.3 (MH +) 520.3 (M + 23); HRMS cale for C2 H44N506 (MH +) 498.3292, found: 498.3309.
Example 55. NI- (2- [2- (benzyloxy) ethyl] carbamoyl-l-methyl-2-oxoethyl) -N 4, N 4 -dimethyl- (2/3) -2-. { [(1/3) -2, 2-dimethyl-l- (neopentylcarboxamido) propyl] carboxamido} butanediamide (88, Table 6). This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was performed by Preparative HPLC. HPLC (system C) 97%, (system D) 95%; IR (KBr)? 3299, 1645, 1527 cm "1; ^ -H-NMR (400 MHz, DMSO-de) d 8.67 and 8.62 (2 xt, J = 5.7 Hz, 1H), 8.15 and 8.10 (2 xd, J = 7.5 and 6.3 Hz, ÍH), 7.98 and 7.93 (2 xd, J = 6.3 and 6.3 Hz, ÍH), 7.61 (d, J = 8.4 Hz, 1H), 7.40-7.20 (m, 5H), 5.0-4.90 (m, 1H ), 4.65-4.55 (m, ÍH), 4.47 (s, 2H), 4.13 and 4.12 (2 xd, J = 8.7 and 8.4 Hz, ÍH), 3.51 (t, J = 6 Hz, 2H), 3.37-3.30 (m, 2H), 2.94 and 2.93 (2 xs, 3H), 2.80 and 2.79 (2 xs, 3H), 2.75-2.60 (m, 2H), 2.20 (d, J = 12.6 Hz, ÍH), 2.04 and 2.02 (2 xd, J = 12.6 y 12.6 Hz, ÍH), 1.23 y 1.22 (2 xd, J = 7.2 y 7.2 Hz, 3H), 0.95 (s, 9H), 0.91 (s, 9H); FAB MS m / z : 604 (MH +), 626 (M + 23); HRMS cale for C3? H5oN5? 7 (MH +) 604.3710, found: 604.3690.
Example 56. Nl-2- [(1,3-benzodioxol-5-ylmethyl) carbamoyl] -1-methyl-2-oxoethyl-N 4, N 4 -dimethyl- (2S) -2-. { [(1/3) -2, 2-dimethyl-l- (neopentylcarboxamido) rovyl] carboxamido} butanediamide (89, Table 6). This compound was prepared according to the procedure for the preparation of α-ketoamides (Example 2). The final purification was carried out by preparative HPLC. HPLC (system C) 99%, (system D) 100%; IR (KBr)? 3302, 1644 cm "1; ^ -H-NMR (400 MHz, DMSO-de) d 9.14 and 9.09 (2 xt, J = 6.4 and 6.1 Hz, ÍH), 8.13 and 8.08 (2 xd, J = 7.5 and 7.5 Hz, 1H), 8.01 and 7.96 (2 xd, J = 6.6 and 6.0 Hz, 1H), 7.60 (d, J = 8.1 Hz, 1H), 6.85-6.80 (m, 2H), 6.73 (d, J = 7.8 Hz, 1H), 5.97 (s, 2H), 5.02-4.86 (m, 1H), 4.65-4.51 (, ÍH), 4.31-4.08 (m, 3H), 2.92 (br s, 3H), 2.80 and 2.79 (2 xs, 3H), 2.74-2.58 (m, 2H), 2.19 (br d, J = 12.6 Hz, ÍH), 2.03 and 2.02 (2 xd, J = 12.9 and 12.6 Hz, 1H), 1.23 (m, 3H), 0.94 and 0.90 (2 xs, 18H); FAB MS m / z: 604 (MH +), 626 (M + 23); HRMS cale for C30H46N5Os (MH +) 604.3347, found: 604.3333.
Example 57. Nl-2- [(1 H -benzo [d 3 imidazol-2-ylmethyl] carbamoyl] -1-methyl-2-oxoethyl-N 4, N 4 -dimethyl- (2/3) -2-. { [(1/3) -2,2-dimethyl-1- (neopentylcarboxamido) rovyl] carboxamido} butanediamide (90, Table 6). This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was carried out by preparative HPLC. HPLC (system C) 93%, (system D) 87%, IR (KBr)? 3294, 1663, 1522 c "1; ifi-NMR (400 MHz, DMSO-de) d 9.41 and 9.35 (2 xt, J = 10.8 and 10.5 Hz, ÍH), 8.64 (, 0.5H), 8.18 and 8.09 (2 xt, J = 14.3 and 15.2 Hz, 2H), 7.99 (d, J = 6.4 Hz, 0.5H), 7.78-7.52 (, 3H), 7.48-7.34 (, 2H), 6.48-6.20 (br d, ÍH) , 5.08-4.93 (m, ÍH), 4.78-4.51 (, 3H), 4.20-4.05 (m, ÍH), 2.96, 2.93, 2.92 and 2.90 (4 xs, 3H), 2.80, 2.79, 2.76, and 2.72 ( 4 xs, 3H), 2.71-2.55 (m, 2H), 2.20 and 2.17 (2 xd, J = 12.4 and 9.8 Hz, ÍH), 2.03 and 2.02 (2 xd, J = 12.7 and 12.7 Hz, ÍH), 1.28 and 1275 (2 xd, J = 7.3 and 7.3 Hz, 2H), 0.95 (s, 9H), 0.94-0.84 (m, 9H); FAB MS m / z: 600.6 (MH +), 618.6 (M + 19); HRMS cale for C3oH 6N706 (MH +) 600.3509, found: 600.3488.
Example 58. N 4, N 4 -dimethyl-N 1 - (1-methyl-2-oxo-2- [(SS) -1-phenylethyl] carbamoylethyl) - (2S) -2-. { [(SS) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (91, Table 6). This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was carried out by preparative HPLC. HPLC (system C) 99%, (system D) 98%; IR (KBr)? 3300, 1647 cm "1; ifi-NMR (400 MHz, DMSO-de) d 9.11 and 9.07 (2 xd, J = 8.6 and 8.6 Hz, ÍH), 8.12 and 8.09 (2 xd, J = 7.9 and 7.3 Hz, ÍH), 8.01 and 7.93 (2 xd, J = 6.4 y 6.3 Hz, ÍH), 7.60 (, ÍH), 7.37-7.17 (m, 5H), 5.01-4.86 (m, 2H), 4.67-4.51 (m, ÍH), 4.14 and 4.13 (2 xd, J = 8.6 y 8.6 Hz, ÍH), 2.93 and 2.92 (2 xs, 3H), 2.79 (br s, 3H), 2.73-2.54 (m, 2H), 2.19 (d, J = 12.7 Hz, ÍH), 2.06 and 2.01 (2 xd, J = 8.3 and 8.3 Hz, ÍH), 1.44-1.37 (m, 3H), 1.24 and 1.17 (2 xd, J = 7.6 and 7.3 Hz, 3H), 0.94 (s, 9H), 0.91 and 0. 90 (2 x s, 9H); FAB MS m / z: 574.4 (MH +), 596.3 (M + 2. 3); HRMS cale for C30H48N5O6 (MH +) 574.3605, found: 574.3586.
Example 59. N 4, N 4 -dimethyl-N 1 - (1-methyl-2-oxo-2- [(IR) -1-phenylethyl] carbamoylethyl) - (2S) -2-. { [(I S) -2, 2-dimethyl-l- (neopentylcarboxamido) propyl] carboxamido} butanediamide (92, Table 6). This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was carried out by preparative HPLC. HPLC (system C) 99%, (system D) 97%; IR (KBr)? 3288, 1645, 1525 cm "1; ^ -H-NMR (400 MHz, DMS0-d6) d 9.11 and 9.05 (2 xd, J = 8.3 and 8.6 Hz, ÍH), 8.13 and 8.07 (2 xd, J = 7.3 and 7.6 Hz, HH), 8.00 and 7.95 (2 xd, J = 6.4 and 6.4 Hz, HH), 7.59 (d, J = 8.6 Hz, 1H), 7.37-7.18 (m, 5H), 5.02-4.84 (m , 2H), 4.63-4.51 (m, ÍH), 4.14 and 4.12 (2 xd, J = 8.6 and 8.6 Hz, ÍH), 2.91 and 2.87 (2 xs, 3H), 2.80 and 2.77 (2 xs, 3H), 2.73 and 2.53 (m, 2H), 2.19 and 2.18 (2 xd, J = 12.7 and 12.4 Hz, ÍH), 2.09-1.98 (m, ÍH), 1.42 (d, J = 7.0 Hz, 3H), 1.23 and 1.19 (2 xd, J = 7.3 and 7.0 Hz, 3H), 0.95 (br s, 9H), 0.91 and 0.90 (2 xs, 9H); FAB MS m / z: 574.4 (MH +), 596.3 (M + 23); HRMS cale for C30H sN5O6 (MH +) 574.3605, found: 574.3591.
Example 60. N 4, N 4 -dimethyl-N 1 - (1-methyl-2-oxo-2- [(IR) -1-phenylpropyl] carbamoyl-ethyl) - (2S) -2-. { [(SS) -2, 2-dimethyl-1- (neopenti carboxamido) propyl] carboxamido} butanediamide (93, Table 6). This compound was prepared according to the procedure for α-ketoamides (Example 2). The final purification was carried out by preparative HPLC. HPLC (system C) 100%, (system D) 96%; GO . { KBr)? 3297, 1647 cm "1; ^ -H-NMR (400 MHz, DMSO-de) d 9.07 and 9.02 (2 xd, J = 8.6 and 8.9 Hz, 1H), 8.13 and 8.06 (2 xd, J = 7.7 and 7.2 Hz, ÍH), 7.99 and 7.94 (2 xd, J = 5.9 and 5.9 Hz, 1H), 7.59 (d, J = 8.3 Hz, ÍH), 7.40-7.18 (m, 5H), 4.98-4.84 (m, ÍH), 4.75-4.65 (m, 1H), 4.63-4.55 (m , 1H), 4.13 (2 xd, J == 8.4 and 8.4 Hz, ÍH), 2.92 and 2.88 (2 xs, 3H), 2.80 and 2.77 (2 xs, 3H), 2.75-2.58 (, 2H), 2.19 and 2.18 (2 xd, J = 12.6 and 12.6 Hz, ÍH), 2.03 and 2.02 (2 xd, J = 12.6 and 12.3 Hz, ÍH), 1.87-1.69 (m, 2H), 1.23 and 1.17 (2 xd, J = 7.2 and 7.2 Hz, 3H), 0.95 and 0.94 (2 xs, 9H), 0.91 and 0.90 (2 xs, 9H), 0.87-0.78 (m, 3H); FAB MS m / z: 588. 7 (MH +), 610.7 (M + 23); HRMS cale for C 31 H 50 N 5 O 6 (MH +) 588.3761, found: 588.3744; Anal (C3? H49N5? 6-H20) C, H, N.
Example 61. Compounds 94 to 98 of Table 7 and 305, 309 and 310 of Table 8 were synthesized according to route (b), Scheme 5. Compounds 301 to 303 and 306 to 308 of Table 8 are synthesized according to the procedure of Example 43. Compound 304 was synthesized according to the procedure of Example 45.
Compound 312 was synthesized in the following way: 312 N4, N-Dimethyl-N1- [1- (R / S) -methyl-2-oxo-3, 3,4,4,4-pentafluorobutyl] - (2) -2-. { 3 - [(3, 3-dimethyl-l-oxobutyl) -amino] -4,4-dimethyl-2-oxopentyl} -butandiamida.
A solution of the intermediate ketomethylene (2.42g, 5.24 mmol, 6,6-dimethyl- (25) -2- (2-dimethylamino-2-oxoethyl) - (55) -5- [(fcer-butoxycarbonyl) amino] - Benzyl 4-oxoheptanoate was prepared according to Moss et al., (J. Med. Chem., 1996, 39, 4173-4180) in 4N HCl / dioxane (30 mL) was stirred at room temperature for 1.5 h. Upon removal of the solvent, the residue was coevaporated twice with CH3CN and then dissolved in CH3CN (50 mL) followed by the addition of i-PrNEt (2.74 mL, 15.72 mmol), tert-butyl acetic acid (0.67 mL, 5.24 mmol) and TBTU (1.70 g, 5.29 mmol) After stirring overnight at room temperature, the solvent was evaporated to dryness, the residue was dissolved in EtOAc and the solution was washed sequentially with 10% aq. NaHC03 sat and brine, and dried (MgSO4). Evaporation to dryness of the solvent gave the desired amide derivative as a brown gummy residue [2.41 g, FAB MS, m / z: 461 (MH +), 483 (M + Na) +]. The crude ida was dissolved in EtOH and the solution was stirred at room temperature in the presence of 10% Pd / C (250 mg) under a H 2 atmosphere for 18 h. After filtering the catalyst and evaporating the solvent to dryness, the oily residue [(1.90 g, FAB MS, m / z: 399 (MH +) corresponding to the ethyl ester derivative] was dissolved in a 1: 1 mixture of MeOH-water (75 L) and solid NaOH was added ( 758 mg) The solution was stirred for 3 h at room temperature and the solvent was evaporated to dryness The residue was dissolved in water, the solution was acidified to pH 2 with 10% aq HCl, extracted with EtOAc and the layers The combined organic extracts were washed with brine, evaporation to dryness of the solvent gave the desired acid as white foam [1.36g, FAB MS, m / z: 371 (MH) + per .C? 9H34N205].
To a solution of the crude acid (370 mg) in CH3CN (50 mL) was added the salt of (3i? / 5, 4. / 5) -4-amino-1, 1, 2, 2-pentafluoro-3 -pentanol • HCl (229 mg, 1 mmol), TBTU (337 mg, 1.05 mmol), i-Pr2NEt (0.70 mL, 4 mmol) and the mixture was stirred at room temperature for 3 h. After evaporation to dryness of the solvent, the residue was dissolved in EtOAc and the solution was washed sequentially with aq. at 10%, NaHCO 3 aq. and brine. The solvent was evaporated to dryness to give the desired product (455 mg, 82% yield). FAB MS, m / z: 546 (MH) + for C 24 H 4 o N 305 F 5.
A cold solution of the hydroxytiamide crude product (454 mg, 0.81 mmol) in EtOAc was oxidized using the Dess-Martin periodinnan (0.69 g, 1.63 mmol). After the usual isolation procedure, the crude product (398 mg) was purified by flash chromatography using a 4: 1 EtOAc-hexane mixture to give the title compound (128 mg). FAB MS, m / z: 544.4 (MH +), 562.4 (M + H20) + by C24H38Ñ305F5.
Example 62. Solid phase synthesis of activated ketones: As shown in the table below, peptidyl trifluoromethyl ketones and α-ketoamides of wide chemical diversity were obtained in 12% -37% total yield of the corresponding starting resin 103 described in Example 1. The raw material, which typically shows a homogeneity of 60-80% by reverse phase HPLC, could be easily purified by semi-preparative HPLC. Since the fragments of trifluoromethyl ketone and α-ketoamide 109 were racemic, the desired inhibitors were usually isolated as a 1: 1 mixture of diastereomers. In some cases each isomer could be separated during purification but in most cases, the inhibitors were subjected to biological tests as a mixture of isomers in the activated ketone center.
Specifically, the compound 218 was synthesized in the following manner: This compound was prepared in solid phase using the resin derived from semicarbazone (103 X '= C (0) NH-Bn). The synthesis in solid phase as well as the cut condition is identical to that reported in example 1. Yield: 33%; HPLC (phosphate): 81%; 1 H-NMR (400 MHz, DMS0-d 6), d 9.18 (t, J = 6.4 Hz, 1 H), 9.14 (s, 1 H), 8.39-8.26 (m, 1 H), 8.08-7.89 (m, 3 H), 7.75 (t, J = 9.5 Hz, 1 H), 7.60 (broad s, 3 H), 7.34-7.22 (m , 6 H), 6.99 (d, J = 8.3 Hz, 2 H), 6.61 (d, J = 8.6 Hz, 2 H), 6.33-6.22 (m, 0.7 H), 4.98-4.94 (m, 2 H) , 4.49-4.43 (, 1 H), 4.33-4.00 (m, 6 H), 3.48 (t, J = 5.8 Hz, 2 H), 2.96-2.91 (m, 1 H), 2.74-2.66 (m, 2 H), 2.05-1.90 (m, 1 H), 1.83 (s, 3 H), 1.65-1.45 (, 4 H), 1.35-1.24 (, 1 H), 1.26 (t, J = 6.4 Hz, 2 H ), 0.98-0.93 (m, 1 H), 0.86-0.82 (m, 6 H); FAB-MS (ES +) cale for C36H52N7O9: 726; found: 726.
The IC50 of compound 218 was found to be 9. 4 μM, Sequence # Performance Cp total (%) 201 Val-Phe-Ser (0-t-Bu) -Asp-Ala (d, l) -CF3 22 202 Val-Phe-Ser (O-t-Bu) -Asp (O-t-14 Bu) Ala (d, l) -CF3 203 Ac-Asn-Asp (O-Bn) -Leu-Ala (d, l) -CF3 40 204 Ph-C (O) Glu-Tyr-Gly-Leu-Ala (d, l) -CF3 68 205 Ac-Phe-Leu-His-Thr-Ala (d, l) -CF3 19 206 Ac-Phe-Leu-His-Thr- (O-tBu) Ala (d, l) - 6 CF3 207 Ac-Gly-Val-Val-Asn-Ala (d, l) -CF3 30 208 Ac-Asp-Glu-Met-Glu-Glu-Abu (d, l) -CF3 36 209 Boc-Gly-Phe-Leu-Abu (d, l) -CF3 23 210 Boc-Val-Ser (O-Bn) -Gly-Asp (O-Bn) -29 Abu (d, l) -CF3 211 Asp (O-Bn) -Ala-Pro-Abu (d, l) -CF3 40 212 Boc-Ala-Ala-Pro-Val (d, l) -CF3 33 213 Ph-CH2-C (O) -Tyr -Ala-Lys-Val (d, i) -CF3 21 214 Ac-Leu-Gly-Asp (O-Bn) -Ala-Val (d, l) - 18 CF3 215 Ac-Gly-Ser (O-Bn) -Leu-Asp (O-Bn) - 18 Vai (d, l) -CF3 216 Ac-Phe-Val-Pro-Val (d or l) -CF3 8 217 Ac-Phe-Val-Pro-Val (d or l) -CF3 1 1 218 Ac-Ser-Tyr-Val-Lys-AIa (d, l) -C (O) -NH- 33 CH2-Ph Ac-Asn-Asp (OBn) -Leu-Ala (d, l) -C (O) -40 NH-CH2-Ph 219 Example 63 ENZYMATIC TESTS Material and Methods: Fluorescent measurements were recorded on a Perkin-Elmer LS-50B spectrofluorimeter equipped with a plate reader accessory. UV measurements were recorded on a Thermomax microplate reader from Molecular Devices. All the specific enzymes and their respective substrates were commercially available with the following suppliers: Boehringer Mannheim (bovine pancreas a-chymotrypsin # 103314 lot 13724423-58, porcine pancreas elastase # 1027891 lot 83260521-23), Calbiochem (human neutrophil elastase # 324681 lot B12778, human liver cathepsin B # 219364 lot B14649, Succ-AAA-pNA # 573459 Lot 510008), Sigma Chemical Co. (Succ-AAPF-pNA # S7388 batch 31H5805, Batch (Z-FR-pNA # L-1242 batch 502774, Succ-AAV-pNA # L-1405, batch 116699).
N0 HCMV protease test: N0 HCMV protease was tested with an internally quenched fluorogenic substrate based on the maturation breakage site (Abz-VVNASSRLY (3-N02) R-OH, kcat / K "= 260 M" is ") . The increase in fluorescence in the Ala-Ser amide bond cut was monitored using the excitation? = 312 nm (aperture 2.5 nm) and emission? = 415 nm (aperture 5 nm). A protocol adaptable to a 96-well plate format was designated-for the determination of IC50 values of inhibitors.
Briefly, the nM HCMV 125 nM protease was pre-incubated for 5 hr at 30 ° C with a range of inhibitor concentrations diluted sequentially (300 to 0.06 μM depending on the potency of each compound). After this period, the enzymatic hydrolysis was initiated by addition of the fluorogenic substrate and the reaction was run for 2 hr at 30 ° C ("30% conversion). It was not required to switch off before the fluorescence measurement since the total search time by the plate reader accessory was relatively short for the duration of the reaction. The incubation buffer (essentially similar to the pre-incubation buffer) contained 50 mM Tris / HCl pH 8, 0.5 M Na2S04, 50 M NaCl, 0.1 mM EDTA, 1 mM TCEP, 3% v / v DMSO and 0.05% casein p / v. The final concentrations of NQ HCMV protease (expressed in terms of total monomer concentration) and substrate were 100 nM and 5 μM respectively. ICso values were obtained by adjusting the inhibition curve to a competitive inhibition model using the SAS NLIN procedure. The inhibition form was determined by measurements of the initial velocities (in test pieces) for several substrates (Abz-Tbg-Tbg-Asn (Me) 2-Ala-SSRLY (3-N02) R-OH) and the concentrations of inhibitors using the same conditions as the previous ones. The results were plotted according to the Cornish-Bowden method ([S] / v vers us [I]) and the Dixon method (1 / v versus [I]) to visually assess the type of inhibition (Cornish-Bowden, A. A simple graphical method for determining the inhibition constants of ixed, uncompetitive and non-competitive inhibitors, Bi ochem J. 1974, 1 37, 143-144).
Specificity tests: The specificity of the compounds was determined against a variety of serine proteases (human leukocyte and porcine pancreatic elastases (HLE and PPE), a-chymotrypsin from bovine pancreas) and a cysteine protease (human liver cathepsin B) . In all cases, a 96-well plate format protocol was used using a specific p-nitroanilide (pNA) colorimetric substrate for each enzyme. Each test included a pre-incubation of 1 hr enzyme-inhibitor at 30 ° C followed by substrate addition and hydrolysis for «30% conversion as measured by looking at a Thermomax UV microplate reader. Substrate concentrations were kept as low as possible compared to KM to reduce substrate competition. The concentrations of the compound ranged from 300 to 0.06 μM depending on its potency. The final conditions for each test were as follows: 50 mM Tris / HCl pH 8, 0.5 M Na2S04, 50 M NaCl, 0.1 mM EDTA, 3% DMSO, 0.01% Tween-20 with [Succ-AAPF-pNA 100 μM. and 250 pM a-chymotrypsin], [Succ-AAA-pNA 133 μM and porcine elastase 8 nM], or [μM Succ-AAV-pNA 133 and 8 nM leukocyte elastase]. 100 mM NaH2P04 pH 6, 0.1 mM EDTA, 3% DMSO, 1 mM TCEP, 0.01% Tween-20, 30 μM Z-FR-pNA and 5 nM cathepsin B (the reserve enzyme was activated in the buffer that contained 20 mM TCEP before use).
Example 64. Biological Results.
Table 1 Compound Structure IC50 (μM) 37 H O Y H O rC ° HNH20 1.8 ± 0.3 o M o o * 43 45 > 300 Table 2 Compound IC50 (μM) 39 46 47 48 52 ± 6 OH 49 HzN '19 ± 3 fifty 51 52 53 54 55 CH3 83 ± 15 56 CH3 > 300 Table 3 Compound 3 IC 50 (μM) 51 2.0 ± 0.3 4. 410.5 57 58 59 3.610.5 60 ± 1 61 Table 4 Compound n R IC50 (μM) 58 1 ± 2 7 AND 1. 62 O 2.8 ± 0.4 66 \ 3.2 ± 0.2 3. 4 + 0.5 1.6 + 0.4 Table 5, ' O J O CH3 Compound z X IC50 (μM) 65 C CF3 l.l ± O.l 74 c CF2CF3 0.11 + 0.01 76 0.20 ± 0.05 79 (OPh) 2 0.66 + 0.06 Table 6.
Compound X IC 5Q (μM) 86 H 1.1 + 0.3 \ ^ NMe C II O 0. 10 + 0.01 Table 7. Cp n R4 Ri IC50 (μM) 97 -r- Me 0.073 Example 65. An interesting compound related to 76 (Table 7), is compound 98 (prepared according to the procedure of Example 2) having the following structure: 98 In the non-HCMV protease test, compound 98 had IC5o = 0.34 μM. The compound with its incorporated iodine atom has the additional benefit of being a useful compound for crystallographic studies of X-rays.
Example 66. Table 8 illustrates more compounds synthesized according to the present invention: Table 8 Compound Structure IC50 (μM) DA-Tbg-Tbg- 308 0.3 Results and Discussion.
After optimization with the peptide portion of the inhibitors, the effect of changes to the activated carbonyl group was considered. This functionality is of particular importance for the inhibition of serine proteases due to the formation of a reversible covalent bond with the serine of the active site. A number of effective activated carbonyl groups have been described in the literature suitable for use with peptidomimetic inhibitors (Mehdi, S. Synthetic and naturally occurring protease inhibitors containing an electrophilic carbonyl group, Bioorganic Chem. 1993, 21, 249-259). Several main classes of these were investigated (Table 6). Compared with trifluoromethyl ketones, the use of pentafluoroethyl ketones, a, a-difluoro-β-ketoamides, α-ketobenzoxazoles, α-ketoamides and diphenyl phosphonates gave significant increases in activity. Inhibitors 74 and 76 showed increases in potency by factors of ten and five respectively.
Several compounds were also investigated to better characterize their interactions with the HCMV protease in terms of the form of inhibition. Figure 1 shows a graph of Dixon obtained for compound 76 which clearly demonstrates that this compound was a competitive inhibitor of HCMV protease.
Compounds 63, 74 and 77 gave similar results indicating that all of these were competitively inhibited (results not shown). It is well known that the interaction of inhibitors based on trifluoromethyl ketone with serine proteases are characterized by a slow onset of inhibition. This phenomenon has been explained by the observation that trifluoromethyl ketones exist in solution almost exclusively in the hydrated form (Edwards, PD, Bernstein, PR Synthetic inhibitors of elastase, Medicinal Research Reviews, 1994, 14, 128-194 and references cited in the same) . This produces a very slow concentration of the inhibitory ketone form and results in time-dependent inhibition. As shown in Figure 2, trifluoromethyl ketone 65 exhibits slow onset of inhibition with an apparent constant rate of 5.4 ± 10 ~ 3 s "l.
Other groups that activate carbonyl were found to be less susceptible to this slow bonding behavior. The progress curve obtained for compound 76, in which equilibrium is reached more rapidly, is shown in Figure 3.
Compound 74 showed intermediate slow bond behavior between 76 and 65, while 77 gave a progress curve comparable to 76. Very slow turnover rate shown by the HMCV protease, coupled with slow binding kinetics for the present series of inhibitors has implications for the formality of enzymatic results. To ensure that the IC50 values obtained were a true reflection of the inhibitory power, test conditions were used in which the inhibitors were pre-incubated with the enzyme before the introduction of the substrate.
Specificity: To evaluate the specificity, the inhibitory activity of the compounds towards a variety of serine proteases was investigated. Compounds 65, 74-85 were tested for inhibitory activity against elastase - porcine pancreas (PPE), human leukocyte elastase (HLE), bovine pancreatic chymotrypsin (BPC), and cathepsin cysteine protease from human liver (cat-B) (results not shown). Compounds 65, 74-79 showed all good specificity profiles against HLE, BPC and cat-B. Some of these compounds were weak PPE inhibitors (which like the HCMV protease showed a preference for alanine in Pi) but with specificity frames of 20 to 300 fold. An important exception to this latter trend is a-ketobenzoxazole 77 which was actually seven times more potent against PPE than against the HCMV protease. A limited SAR of benzoxazole substitutions was carried out to try to improve the specificity profile of these compounds. Benzothiazole 80 proved to be a potent inhibitor of HCMV (IC50 1.1 μM) and also strongly interacted with PPE (IC50 9 μM). Compound 81 was not an inhibitor of PPE but this enhanced specificity was accompanied by an 18-fold loss in activity towards the HCMV protease. Several methylated benzoxazoles 82-85 were more potent inhibitors of PPE than of HCMV protease.
Compound 76 represented one of the most potent HCMV protease inhibitors described thus far. This structure also suggested the possibility of further increasing the potency by extending the C-terminal amide radical of this inhibitor into the Si 'bond cavity of the enzyme. The observation that Pi 'conserved amino acids clearly (alanine or serine) suggested extending the C-terminus of the a-ketoamide class of inhibitors to try to take advantage of the interactions in the S' cavity.
To further improve the potency of compound 93, the extension in residue P4 was applied in the Pl series of glycine and alanine (Table 7). Since the glycine and alanine series are equipotent, the following observations can be made. The incorporation of a terminal amine in the P4 residue results in a 5-fold loss in potency while the addition of a Boc group in this amine gives a 9-fold improvement in potency. Further extensions at residue P4 in the form of a group covering 6-aminocaproyl protected with Boc gave compound 96 which had an IC50 value of 75 nM. Removal of the Boc group from this inhibitor improved the potency by a factor of 2 to give compound 97 which is less than 40 nM in potency and represents the most potent compound in this series.
Table 8: Compounds 301 to 312 summarize different substitutions of the P2 side chain that gave potent inhibitors. These include several amide substitutions of asparagine and a new sulfonamide residue.
Table 9: Compounds 401 summarize different substitutions in Pl *.
Table 9 Cp # Structure IC5o (μ M) 413 *? R A tr 4.4 It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.
Having described the invention as above, the content of the following is claimed as property.
LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: BOEHRINGER INGELHEIM (CA ADÁ) LTD (B) STREET: 2100 CUNARD STREET (C) CITY: LAVAL (D) STATE: QUEBEC (E) COUNTRY : CA ADÁ (F) ZIP CODE: H7S 2G5 (G) TELEPHONE: (514) 682-4640 (H) TELEFAX: (514) 682-8434 (ii) TITLE OF THE INVENTION: PEPTIDOMYMETIC INHIBITORS OF PROTEASE OF CITOMEGALIVIRUS OF HUMAN (iii) SEQUENCE NUMBER: 72 (iv) COMPUTER READING FORM: (A) TYPE OF MEDIA: Flexible disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) PACKAGE: Patentln Relay # 1.0, Version # 1.30 (EPO) (v) CURRENT APPLICATION DATA: APPLICATION NUMBER: (vi) DATA FROM THE PREVIOUS APPLICATION: (A) APPLICATION NUMBER: US 60 / 034,041 (B) DATE OF SUBMISSION: 27-DEC-1996 (vi) DATA FROM THE PREVIOUS APPLICATION: (A) NUMBER OF APPLICATION: US 60 / 052,860 (B) DATE OF SUBMISSION: JULY 17, 1997 (vi) DATA FROM THE PREVIOUS APPLICATION: (A) NUMBER OF APPLICATION: US 60 / 059,806 (B) DATE OF SUBMISSION: 23-SEP-1997 (2) INFORMATION OF SEQ ID NO: 1 i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note = wN-terminal is covered with acetyl " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 1 Gly Val Val Asn Ala 1 5 (2) INFORMATION OF SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) L0CALIZATION: 1 (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "Xaa in position 5 is [d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 2 Gly Val Val Asn Xaa 1 5 (2) INFORMATION OF SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "Xaa in position 5 is (d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note »" C-terminal is modified with activated carbonyl: C (0) CF3"(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 3: Ser Trp Val Lys Xaa 1 5 (2) INFORMATION OF SEQ ID NO: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 1 (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 4: Ser Trp Val Lys Ala 1 5 (2) INFORMATION OF SEQ ID NO: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 5: Val Val Asn Ala 1 (2) INFORMATION OF SEQ ID NO: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Xaa in position 4 is (d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 6: Val Val Asn Xaa 1 [2) INFORMATION OF SEQ ID NO: 7 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCA IZATION: 4 (D) OTHER INFORMATION: / nota = "Xaa in position 4 is (d) Abu ((d) Abu is acid residue of amino acid (R) -2-aminobutyric) " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 7: Val Val Asn Xaa (2) INFORMATION OF SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = w -terminal is covered with acetyl " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / nota = "Xaa in position 4 is Abu (amino acid residue of (S) -2-aminobutyric acid)" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 8: Val Val Asn Xaa 1 (2) INFORMATION OF SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Xaa en position 4 is (d) Apn; (d) Apn = (R) -2-amino pentanoic acid " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 9: Val Val Asn Xaa 1 [2) INFORMATION OF SEQ ID NO: 10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Xaa in position 4 is Apn; Apn = acid (S) -2-amino pentanoic" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 10: Val Val Asn Xaa 1 (2) INFORMATION OF SEQ ID NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 11 Val Val Gln Ala 1 (2) INFORMATION OF SEQ ID NO: 12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Xaa in position 4 is ¡d) Alanina" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 12: Val Val Gln Xaa 1 [2) INFORMATION OF SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) L0CALIZATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 13: Val Val Asp Ala 1 [2) INFORMATION OF SEQ ID NO: 14: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear ; ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note = "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCA IZATION: 4 (D) OTHER INFORMATION: / note = "Xaa in position 4 is (d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14: Val Val Asp Xaa 1 (2) INFORMATION OF SEQ ID NO: 15: ;) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15: Val Val Ser Ala 1 2) INFORMATION OF SEQ ID NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 16: Val Val Ser Xaa 1 (2) INFORMATION OF SEQ ID NO: 17: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCA IZATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 17: Val Val Lys Ala 1 (2) INFORMATION OF SEQ ID NO: 18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is; d) alanine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 18: Val Val Lys Xaa 1 '2) INFORMATION OF SEQ ID NO: 19: i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is beta- (2-thiazolyl) alanyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 19: Val Val Xaa Ala (2) INFORMATION OF SEQ ID NO: 20: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is beta- (2-thiazolyl) alanyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 20: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 21: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) L0CALIZATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 21 Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 22 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 22: Val Val Xaa Xaa 1 [2) INFORMATION OF SEQ ID NO: 23: [i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" [xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 23: Val Val Leu Ala 1 (2) INFORMATION OF SEQ ID NO: 24 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" ; ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 24 Val Val Leu Xaa 1 [2) INFORMATION OF SEQ ID NO: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 25: Val Val Phe Ala 1 2) INFORMATION OF SEQ ID NO: 26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) Alanine " (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 26: Val Val Phe Xaa 1 (2) INFORMATION OF SEQ ID NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 27: Val Val Val Ala 1 (2) INFORMATION OF SEQ ID NO: 28: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear ; Ü) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 28: Val Val Val Xaa 1 (2) INFORMATION OF SEQ ID NO: 29: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCA IZATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 29: Val Val Ala Ala 1 (2) INFORMATION OF SEQ ID NO: 30: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEC ID NO: 30: Val Val Ala Xaa 1 (2) INFORMATION OF SEQ ID NO: 31: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" ; ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 D) OTHER INFORMATION: / note- "Xaa in position 3 is (d) Alanine " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 31: Val Val Xaa Ala (2) INFORMATION OF SEQ ID NO: 32: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is (d) Alanine " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEC ID NO: 32 Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 33: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCALIZATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Abu " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 33: Val Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear ; ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Abu " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 34: Val Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 35: : i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 35: Val Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 36: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear ; ÍÍ) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg * (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 36: Val Xaa Xaa Xaa 1 [2) INFORMATION OF SEQ ID NO: 37: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) L0CA IZATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is methyl leucine" ; ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 37: Val Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 38: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION:! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is methyl leucine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 38: Val Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 39: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCA IZACIÓN :! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is (1-adamantyl) glycine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 39: Val Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 40: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is (1-adamantyl) glycine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 40: Val Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 41: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Dimethyl Asian acid" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCA IZATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 41: Val Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 42: Ü) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is dimethyl aspartic acid" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCA IZATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEC ID NO: 42: Val Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 43: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Xaa in position 1 is dimethyl glycine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCALIZATION; 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 43: Xaa Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 44: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide ; ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "Xaa in position 1 is dimethyl glycine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg ' (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) CF3" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 44: Xaa Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 45: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION:! (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg " (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with C (O) NHCH (CH2CH3) phenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 45: Xaa Xaa Xaa Ala 1 [2) INFORMATION OF SEQ ID NO: 46: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! ~ ~ "" "(D) OTHER INFORMATION: / note-" Xaa in position 1 is Tbg " (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg ' (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with C (O) NHCH (CH2CH3) phenyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 46: Xaa Xaa Xaa Xaa (2) INFORMATION OF SEQ ID NO: 47: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Amino covered with Boc residue" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg ' (ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg "(ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note-" Xaa in position 3 is dimethyl asparagine " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with C (O) HCH (CH2CH3) phenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 47: Xaa Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 48: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Amino covered with Boc residue" (ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg " (ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal is modified with C (O) NHCH (CH2CH3) phenyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 48 Xaa Xaa Xaa Xaa (2) INFORMATION OF SEQ ID NO: 49: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Amino covered with Boc-NH (CH2) 5-C (0)" [ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION.-l (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg ' (ix) FEAT (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carbonyl group replaced by C (O) H (CH2CH3) phenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 49: Xaa Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 50: [i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Amino covered with Boc-NH (CH2) 5-C (0) " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carbonyl group replaced by C (O) NH (CH2CH3) phenyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 50: Xaa Xaa Xaa Xaa (2) INFORMATION OF SEQ ID NO: 51: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION:! (D) OTHER INFORMATION: / note- "Amino covered with H2N (CH2) 5C (0)" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carbonyl group is replaced by C (O) HCH (CH2CH3) phenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 51: Xaa Xaa Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 52: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Amino covered with H2N (CH2) 5C (0)" (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "Xaa in position 1 is Tbg " (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 2 (D) OTHER INFORMATION: / note- "Xaa in position 2 is Tbg " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carbonyl group is replaced by C (O) HCH (CH2CH3) phenyl" ( ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is [d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 52: Xaa Xaa Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 53: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa en position 3 is dimethyl asparagine " (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "carboxyl group in C-ter inal is replaced by C (O) HCH2C (CH3) 3" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 53: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 54: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NHCH2C (CH3) 3" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) Alanine " (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 54 Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 55: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note- "C-terminal carbonyl group is replaced by OCH2Penyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4..5 (D) OTHER INFORMATION: / note- "Carboxi added between Ala in position 4 and Ala in position 5" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 55: Val Val Xaa Ala Ala 1 5 (2) INFORMATION OF SEQ ID NO: 56: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note- "Carbonyl group in C-terminal is replaced by OCH2Penyl" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4..5 (D) OTHER INFORMATION: / note- "Added carboxy function between (d) Ala in position 4 and Ala in position 5" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 56: Val Val Xaa Xaa Ala 1 5 (2) INFORMATION OF SEQ ID NO: 57: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) HCH2 (4-trifluoromethylphenyl)" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 57 Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 58: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (0) NHCH2 (4-trifluoromethylphenyl)" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) Alanine " (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 58: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 59: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal covered with Boc residue" " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" ; ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NH (4-methylphenyl)" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 59: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 60: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal covered with Boc residue" " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NH (4-methylphenyl)" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 60: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 61 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NHphenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 61 Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 62: li) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "carboxyl group in C-ter inal is replaced by C (O) Hphenyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 62: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 63: [i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) Hcyclohexyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 63: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 64: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NH-cyclohexyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 64: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 65: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (0) NHCH2CH2phenyl" [xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 65: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 66: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION; / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "carboxyl group in C-terminal is replaced by C (0) NHCH2CH2phenyl " (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) L0CALIZATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 66: Val Val Xaa Xaa (2) INFORMATION OF SEQ ID NO: 67 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) HCH2cyclohexyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 67: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 68: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCALIZATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (0) NHCH2cyclohexyl" (x) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "Xaa in position 4 is (d) alanine" [xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 68: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 69: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION:! (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "carboxyl group in C-terminal is replaced by C (O) H- (3-amidophenyl) " (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 69: Val Val Xaa Ala 1 (2) INFORMATION OF SEQ ID NO: 70: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCALIZATION: 1 (D) OTHER INFORMATION: / note- "N-terminal is covered with Boc" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 3 (D) OTHER INFORMATION: / note- "Xaa in position 3 is dimethyl asparagine" (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note- "C-terminal carboxyl group is replaced by C (O) NH- (3-amidophenyl) "(ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION: 4 (D) OTHER INFORMATION: / note-" Xaa in position 4 is (d) alanine " (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 70: Val Val Xaa Xaa 1 (2) INFORMATION OF SEQ ID NO: 71 [i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (Ü) TYPE OF MOLECULE: peptide (ix) FEATURE: (A) NAME / KEY: Modified site (B) LOCATION :! (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (0) -NH-CH2-phenyl" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 71 Ser Tyr Val Lys Ala 1 5 (2) INFORMATION OF SEQ ID NO: 72: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTIC: (A) NAME / KEY: Modified site (B) LOCATION: l (D) OTHER INFORMATION: / note- "N-terminal is covered with acetyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note- "C-terminal is modified with activated carbonyl: C (O) -NH-CH2-phenyl" (ix) CHARACTERISTICS: (A) NAME / KEY: Modified site (B) LOCATION: 5 (D) OTHER INFORMATION: / note- "Xaa in position 5 is [d) alanine" [xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 72: Ser Tyr Val Lys Xaa 1 5

Claims (23)

1. A compound of formula I: (I) characterized in that z is C or P; when z is C, then X is CF3; C2F5; benzothiazole; oxazolo [4, 5b] pyridine; or benzoxazole-R7 wherein R7 is H or methyl; OX is CF2C0NH-R6, C (0) NH-R6, where e is Co-io alkyl optionally substituted with phenyl or cyclohexyl, the phenyl ring or cyclohexyl is optionally substituted with Me, halogen, -CF3, -CH (Me ) -C (O) - OBn; -C (0) NH2; or -C (0) -morpholino; the phenyl or cyclohexyl ring is optionally fused with a phenyl ring; (CH2)? _3-0- (CH2)? _3-phenyl The phenyl is optionally substituted with halogen; (CH2)? _3- 2 -ben z imide zol; (CH2) 1-3- (3,4-methylenedioxybenzene); or (CH2)? _ 3-0-C (0) -0CH2CH = CH2; or, when z is P, then X is - OPh) 2 Ri is H, Me, or Et; R2 is CH2-S02NH2; alkyl -C? _6; - (C? _6 alkyl) aryl; - (C 1-6 alkyl) thiazole; -CH2C (0) - (Ci-β alkyl); CH2C (0) -pyrrolidino; -CH2C (0) -morpholino; - (C? _6 alkyl) amino; - (C? _ Alkyl) amido optionally mono- or disubstituted with Ci-β alkyl; alkyl is optionally substituted with pyridino; W is NH, CH2 or CH (CH3); R 3 is alkyl 1-12 r C 1-6 alkyl) C (0) OH; or adamant ilo; n is 0 or 1, R 4, when n is 1, is alkyl -Ci-β or - (C? -6 alkyl) -aryl wherein the aryl is optionally substituted with OH; is 0 or 1, Rs, when m is 1, is H or -CH 2 OH; Y And it is H; (CH2) 2-t-Bu; or an acyl of the formula: -C (0) - (CH 2)? - 6-C (0) OH; -C (O) - (CH2)? -6 -Ph where Ph is optionally substituted with OH; -C (0) -CH2N (CH3) 2; -C (0) -R9; -C (0) 0-R9; or -C (0) NH-R9 wherein R9 is C6_6 alkyl; or -C (O) - (CH2)? -6-NH2 wherein the amino group is optionally protected with an amino protecting group.
2. A compound according to claim 1, characterized in that z is C; X is CF3; C2F5; 2-benzothiazole; 2-oxazolo [4, 5b] pyridine; 2-benzoxazole-R, wherein R is H, 4-Me, 5-Me, 6-Me, or 7-Me; CF2CONHR6 or C (0) NHR6 wherein e is C? - alkyl, optionally substituted with cyclohexyl, naphthyl, or phenyl optionally substituted with Me, iodo, CF3, -CH (Me) -C (0) -OBn; -C (0) NH2, or C (O) -morpholino; (CH2) 2-0-CH2-phenyl; CH2-2-benzimidazole; or CH2- (3,4-methylenedioxybenzene); when z is P, then X is (OPh) 2. Ri is H, methyl or ethyl R2 is -CH2- (4-thiazolo); - (CH2)? _ 4-NH2; -CH2-C (O) -ter-butyl; -CH2-C (O) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); -CH2S02NH2; - (CH2)? -2-ami or, the amide nitrogen optionally mono- or di-substituted with a substituent independently selected from: CH3; t-Bu; phenyl; or -CH2CH2- (2-pyridino); W is NH or CH2; R3 is ethyl; isopropyl; t-Bu; CH2-t-Bu; or adamantyl; n is 0 or 1; R4, when n is 1, is isopropyl; t-Bu; or 4-hydroxybenzyl; m is O or l; R5, when m is 1, is H; And it is H; -CH2-CH2- t-Bu; or an acyl of formula -C (0) CH3; -C (0) CH2-CH (CH3) 2; -C (0) CH2-t-Bu (DA-Tbg); -C (O) (CH2) 2-4-hydroxyphenyl; -C (O) - (CH2) 3-C00H; -C (O) O-t-Bu (Boc); -C (O) H-t-Bu; -C (0) CH2-N (CH3) 2; or -C (O) (CH2)? 6NH2, the amino group is optionally protected with an amino protecting group.
3. A compound according to claim 2, characterized in that z is C; X is CF3; C2F5; 2-benzothiazole; 2-benzoxazole-R7, wherein R7 is H, 4-Me, 5-Me, 6-Me, or 7-Me; -CF2C0NH-CH2-phenyl; -C (0) NHR6 wherein R6 is -CH (Me) (CH2) 4CH3; cyclohexyl; naphthyl; -CHZ-phenyl; -CH (CH 3) -phenyl; or -CH (CH2CH3) -phenyl; -CH2-4-iodophenyl; -phenyl-CH3; -phenyl-CF3; -f-enyl-C (O) NH2; phenyl-C (O) -orfolin; -f-enyl-CH (Me) -C (O) -OBn; - (CH2) 2-0- CH2-phenyl; -CH2-2-benzimidazole; -CH2- (3,4-methylenedioxybenzene); or - (CH2) 2-0-C (O) -OCH2CH = CH2; or when z is P, then X is (OPh) 2 Ri is H or methyl; R2 is -CH2-C (O) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); -CH2S02NH2; - (CH2) C (0) NH2; - (CH2) 2C (0) N (CH3) 2; -CH2-C (0) -NH-t-Bu; or - (CH2) 2-C (0) -N (CH3) CH2CH2 (2-pyridino); W is NH; R3 is ethyl; isopropyl; or t-Bu, R4, when n is 1, is isopropyl; or t-Bu; Rs, when m is 1, is H; And it is H; or an acyl of formula -C (0) CH3; -C (O) CH2-CH (CH3) 2; -C (0) CH2-t-Bu (DA-Tbg); -C (O) (CH2) 2-4-hydroxyphenyl; -C (O) - (CH2) 3 -COOH; -C (0) 0-t-Bu (Boc); -C (O) (CH2) 5 NH2; or
4. A compound according to claim 3, characterized in that z is C; X is C2F5; -C (0) NHR6 wherein Re is -CH2-phenyl; -CH2-4-iodophenyl; CH (CH 3) -phenyl; or -CH (CH2CH3) -phenyl; CH (Me) -naphthyl; -CH2CH (Me) -phenyl; - (CH2) 2-0- CH2-phenyl; -CH2-2-benzimidazole; or -CH2- (3,4-methylenioxybenzene); Ri is H or methyl; R2 is -CH2-C (O) - (N-pyrrolidino); -CH2-C (O) - (N-morpholino); - (CH2) 2C (0) N (CH3) 2; or - (CH2) 2-C (O) -N (CH3) CH2CH2 (2-pyridino); It's NH. R3 is isopropyl; or t-Bu; R4, when n is 1, is t-Bu; m is 0, and And it is an acyl of formula: -C (0) CH2-t-Bu (DA-Tbg); -C (0) 0-t-Bu (Boc); -C (O) (CH2) 5 NH2; or -C (O) (CH2) 5NH-B0C
A compound of formula characterized in that X is CF3, C2F5, 2-benzothiazole, CF2CONHR6, CONHR6, wherein R6 is CH2C6H5, CH2 (4-iodophyl), CH3, (CH2) 2OCH2C6H5, CH2-2-benzimidazole, CH2- (3,4-methylenedioxybenzene), CH (CH3) C6H5 or CH (CH2CH3) C6H5; or X is 2-benzoxazole-R7 wherein R is H, 4-CH3, 5-CH3, 6-CH3 or 7-CH3; Ri is H, CH 3 or CH 2 CH 3; R2 is CH2C0NH2, CH2CH2C0NH2, CH2-thiazole, CH2C0N (CH3) 2, CH2C0- (pyrrolidino); R3 is Et, CH (CH3) 2, C (CH3) 3, adamantyl, CH2C (CH3) 3 or C (CH3) 2C02H; R20 is COCH2C (CH3) 3, COCH2CH2C6H4OH, COCH2CH (CH3) 2, C02C (CH3) 3, CONHC (CH3) 3, C0CH2N (CH3) 2, CO (CH2) 3C02H, CO- fS -CH (NH2) C ( CH3) 3, CO- (S) -CH. { NHC (O) O-C (CH3) 3.}. C (CH3) 3, CO-f-S-CH. { NHCO (CH2) 5NHC (0) OC (CH3) 3} C (CH3) 3 or CO- (S) -CH. { NHCO (CH2) 5NH2} C (CH3) 3.
6. A compound according to claim 1, characterized in that it is selected from the group consisting of: NI- (3,3,3-trifluoro-1-met i 1-2 -oxopropyl) - (2S) -2- ((IS) -2-methyl-1 - [((SS) -2-methyl-1) - [(Methylcarboxamido) methyl] carboxamidopropyl) carboxamido] propylcarboxamido) butanediamide (37) NI- (3, 3, 3-trifluoro-1-methyl-1-2-oxopropyl) - (25) -6-amino-2- ((1) -1 - [((15) -1 - [(15) -2-hydroxy-1- (methylcarboxamido) ethyl] carboxamido-2- (4-hydroxyphenyl) ethyl) carboxamido] -2-methylpropylcarboxamido) hexanamide (38); NI- (3, 3, 3-trifluoro-1-met i 1-2 -oxopropyl) - (2S) -2- [((15) -2-methyl-l- [(15) -2-methyl-1 - (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (39); NI- (3, 3, 3-trifluoro-1-methy1-2-oxopropyl) - (2S) -2-. { (15) -2-methyl-l- [(methylcarboxamido) -propyl] carboxamido} butanediamide (40); NI- (3, 3, 3-trifluoro- (15) -met i 1-2 -oxopropyl) - (25) -2-. { (15) -2-methyl-l- [(methocarboxamido) propyl] carboxamido} butanediamide (43); Nl- (l-ethyl-3,3,3-trifluoro-2-oxopropyl) - (25) -2- [((15) -2-methyl-l- [(15) -2-methyl-1- ( methylcarboxamidopropyl] carboxamidopropyl) carboxamido] butanediamide 44 NI - (1- (3, 3, 3, -trifluoro-l-propyl-2-oxopropyl) - (25) -2- [((15) -2-methyl-l- [(IS) -2-methyl) -l- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (45); NI- (3, 3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2- [((15) -2-methyl-1 - [(15) -2-methyl-1- ( methylcarboxamido) propyl] carboxamidopropyl) carboxamido] pentanediamide (46); Acid (35) -3 - [((15) -2-methyl-l- [(15) -2-methyl-1- (methylsarboxamido) propyl] carboxamido-propyl) carboxamido] -3- [(3, 3, 3-trifluoro-1-methyl-2-oxopropyl) carbamoyl] propanoic (47); Nl - f (15) - l - ((15) -2 -hydroxy- 1- [(3,3, 3-trifluoro-1 -met i1-2-oxopropyl) carbamoyl] ethyl-carbamoyl) -2-met i lpropi1] - (25) -3-methyl 1-2- (methylcarboxamido) butanamide (48); NI - (3, 3, 3-trifluoro-1-methyl-2-oxopropy1) - (25) -6-amino-2- [((15) -2-methyl-l- [(15) -2-methy] 1-1- (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] hexanamide (49); NI - [(15) -2-methyl-l- ((15) -2- (1,3-thiazol-4-yl) -l - [(3,3,3-trifluoro-1-met i 1- 2-oxopropyl) -carbamoyl] ethylcarbamoyl) propyl] - (25) -3-met i 1-2- (methylcarboxamido) butanamide (50); N4, N4 -dimet il -Nl- (3,3, 3-t-rifluoro-1-methy1-2-oxopropyl) - (25) -2- [((15) -2-methyl-l- [(15) -2-methyl-1- (methylcarboxamido) propyl] boxamidopropyl) carboxamido] butanediamide (51); NI - (3, 3, 3-trifluoro-1-met il-2-oxopropi 1) - (2S) -4-methyl-2- [((15) -2-methyl-l- [(15) -2 -met-il-1- (methylcarboxamido) propyl] carboxamidopropyl 1) carboxamido] pentanamide (52); NI - [(15) -2-methyl-l- ((15) -2-phenyl-l- [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) propyl] - (25) -3-methyl-2- (methylcarboxamido) butanamide (53); N4, N4 -dimet il -Nl- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2 - [((15) -l - [(15) -2-methi 1 -1 - (methylcarboxamido) propyl] carboxamidopropyl) carboxamido] butanediamide (57); N4, N4 -dimet il -Nl- (3, 3, 3-trifluoro-1-met il-2-oxopropyl) - (25) -2- [((15) -2, 2-dimeti1-1- [( 15) -2-methyl-1- (methylcarboxamido) propyl] carboxamido-propyl) carboxamido] butanediamide (58); N4, N4 -dimet il -Nl- (3,3, 3-trifluoro-1-met il-2-oxopropyl) - (25) -2 - [((15) -3,3-dimethyl-l- [( 15) -2-met il-1- (methylcarboxamido) propyl] carboxamido butyl) carboxamido] butanediamide (59); N4, N4 -dimet il -Nl- (3,3, 3-trifluoro-1-methyl-1-2-oxopropyl) - (25) -2- [((5) -l- (1 -admant il) -1 - [(1S) -2- et il-1- (methylcarboxamido) propyl] carboxamido methyl) carboxamido] utanediamide (60); Acid (35) -3- ((15) -2- (dimet i Icarbamoyl) -l- [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] -ethylcarbamoyl) -2,2- dimet i 1-3- [(15) -2-methi 1-1- (methylcarboxamido) propi 1] carboxamidopropanoic acid (61); N4, N4 -dimet il -Nl- (3,3, 3-trifluoro-1-met il-2-oxopropyl) - (25) -2 - [(15) -2, 2 -dimethyl-1- (meth. 1 carboxamido) propyl] carboxamidobutanediamide (62); N4, N4 -dimet il-Nl- (3, 3, 3-trifluoro-1-methyl 1-2-oxopropyl) - (25) -2 - ((15) -l- [(4-hydroxyphenyl-ethyl) carboxamide ] -2, 2-dimethylpropi 1carboxamido) butanediamide (63); N4, N4-dimethyl-Nl- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2- [(15) -l-. { isobutylcarboxamido) -2,2-dimethylpropyl] carboxamidobutanediamide (64); N4, N4 -dimet il-Nl- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2 - [(15) -2, 2-dimethyl-l- (neopentylcarboxamido) propyl ] carboxamidobutandiami a (65); N4, N4 -dimet il-N1- (3,3, 3-trifluoro-1-methyl-2-oxopropyl) - (25) -2- ((15) -1- [(3,3-dimethyl-1-butyl) amino] -2,2-dimethylpropycarboxamido] butanediamide (66); 4 ?, 4? -Dimethyl-1? - (3,3, 3-trifluoro-1-methyl-2-oxopropyl) -2- [1- (er-butoxycarbonyl-amino) -2, 2-dimethyl-il- ( ÍS) -propylcarboxamido] - (2S) -butanediamide (67); N4, N4-Dimethyl-? L- (3,3, 3-trifluoro-1-methyl-2-oxopropyl-2- [1- (tert-butylaminocarbonyl-amino) -2,2-dimethyl- (SS) -propylcarboxamido ] - (2S) -butanediamide (68); N4, N4 -dimet i l -Nl- (3,3, 3-trifluoro-1-methy1-2-oxopropyl) - (25) -2- [. { (15) -1- [(dimethylamino) methyl] carboxamido-2,2-dimethylpropy1) carboxamido] butanediamide (69); 4- [(15) -l- ((lS) -2- (dimethylcarbamoyl) -1 - [(3,3,3-trifluoro-l-methyl-2-oxopropyl) carbamoyl] ethylcarbamoyl) -2, 2 -dimeti lpropi1] carbamoylbutanoic (70); N4, N4 -dimet il -Nl- (3,3,4,4, 4 -pentafluoro-1-methyl-2-oxobutyl) - (25) -2 - [(15) -2, 2-dimeti 1-1 - (neopentyl carboxamido) propyl] carboxamidobutanediamide (74); NI - [3- (benzylcarbamoyl) -3,3-difluoro-1-methyl-2-oxopropyl] -N4, N4 -dimethyl- (25) -2 - [(15) -2, 2-dimet i 1-1 - (neopentylcarboxamido) propyl] carboxamidobutanediamide (75); Benzyl acid amide 3-. { 2- [2- (3, 3-Dimethyl-butyrylamino) -3,3-dimethyl-butyrylamino] -3-dimethylcarbamoyl-propionylamino} -2-oxo-butyric (76); NI- [2- (1, 3-benzoxazol-2-yl) -1-met i 1-2 -oxoethyl] -N 4, N 4 -dimethyl- (25) -2-. { [(15) -2, 2 -dimet i 1-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (77); Diphenyl N4, N4 -dimet il-Nl- (1-aminoethylphosphinate) (25) -2-. { [(15) -2,2-dimethyl-l- (neopentylcarboxamido) propyl] carboxamido} butan diamide (79); NI- [2- (1, 3-benzothiazol-2-yl) -1-met i 1-2 -oxoethyl] -N 4, N 4 -dimethyl- (25) -2-. { [(15) -2, 2 -di-methyl-1- (neopentylcarboxamido) propyl] carboxamido} butan diamide (80); N 4, N 4 -dimet il-N 1 - (l-methyl-2- [1,3] oxazolo [4,5-] pyridin-2-yl-2-oxoethyl) - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (81); N 4, N 4 -dimethyl- NI- [l-methyl-2- (6-methyl-1-1, 3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimet i 1-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (82); N 4, N 4 -dimet il-N 1 - [l-methyl-2- (5-methi 1-1, 3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimeti 1-1- (neopent i 1carboxamido) propyl] carboxamido} butanediamide (83); N4, N4 -dimethyl- NI- [l-methyl-2- (4-methyl-1,3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimethy1-1- (neopentylcarboxamido) rovyl] carboxamido} butanediamide (84); N 4, N 4 -dimet il-N 1 - [l-methyl-2- (7-met il-1,3-benzoxazol-2-yl) -2-oxoethyl] - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (85); N4, N4 -dimet il-Nl- [l-methyl-2- (meth i Icarbamoyl-2-oxoethyl] - (25) -2- { [(15) -2, 2-dimethyl-l- (neopentylcarboxamido ) propyl] carboxamido.} butanediamide (86); Nl- (2- [2- (benzyloxy) ethyl] carbamoyl-l-methyl-2-oxoethyl) -N 4, N 4 -dimethyl- (25) -2-. { [(15) -2,2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (88); Nl-2- [(1,3-benzodioxol-5-ylmethyl) carbamoyl] -1-methyl-2-oxoethyl-N4, N4-dimethyl- (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (89); Nl-2- [(1 # -benzo [d] i ida zol-2-ylmethyl) carbamoyl] -1-methyl-2-oxoethyl-N 4, N 4 -dimethyl- (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) rovyl] carboxamido} butanediamide (90); N 4, N 4 -dimet il-N 1 - (1-methy1-2-oxo-2- [(SS) -1-phenylethyl] carbamoylethyl) - (25) -2-. { [(15) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (91); N 4, N 4 -dimet il-N 1 - (l-methyl-2-oxo-2- [(li?) -1-phenilethyl] carbamoylethyl) - (25) -2-. { [(15) -2, 2-dimeti 1-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (92); N4, N4-dimethyl-Nl- (1-methyl-2-oxo-2- [(li?) -1-phenylpropyl] carbamoyl-yl) - (25) -2-. { [(1S) -2, 2-dimethyl-1- (neopentylcarboxamido) propyl] carboxamido} butanediamide (93); 94; 95; 97; Ac-Ser-Tyr-Val-Lys-Ala (d, 1) -C (O) -NH-CH2-Ph 218; 303; 307; 311; 401; 402; 403, 404; 405; 408; 413; Y 414.
The compound according to claim 6, characterized in that it is selected from the group consisting of: compound number 37, 38, 39, 44, 46, 50, 51, 53, 57, 58, 59, 60, 62, 63, 64 , 65, 66, 67, 68, 69, 70, 74, 75, 76, 77, 79, 80, 82, 83, 84, 85, 86, 88, 89, 90, 91, 92, 93, 94, 95 , 96, 97, 98, 218, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, and 401 to 414.
The compound according to claim 7, characterized in that it is selected from the group consisting of: compound number 37, 51, 58, 63, 64, 65, 70, 74, 75, 76, 77, 79, 80, 82, 83 , 84, 85, 86, 88, 89, 90, 92, 93, 94, 95, 96, 97, 98, 304, 305, 306, 307, 308, 309, 310, 311, 312, 401, 403, 404 , 405, 406, 407, 408, 409, 410, 411, 412, and 414.
The compound according to claim 8, characterized in that it is selected from the group consisting of: compound number 74, 76, 88, 89, 90, 92, 93, 95, 96, 97, 98, 305, 308, 309, 407 , and 408.
10. A solid phase process for the synthesis of peptidyl-activated ketones, characterized in that it comprises the step of: a) coupling a semicarbazone acid of formula 113 to a resin by i n activation if; wherein R is a side chain of a natural or non-natural amino acid; and X * is CF3, CF2CONH-R3o, C (0) NH-R30, or C (0) OR30, wherein R30 is a cyclic C3_i2 alkyl or C? -? or acyclic alkyl or cyclic C3_? 2 alkenyl or C2_ alkenyl? Acyclic, the alkyl or alkenyl is optionally substituted with NH 2, OH, SH, halo, or carboxyl; the alkyl or alkenyl optionally contains at least one heteroatom independently selected from the group consisting of: O, S, and N; or R 30 is a C 1 or C 1 aralkyl aryl or optionally substituted with C 1 -C 6 alkyl, NH 2, OH, SH, halo, carboxyl or carboxyalkyl (lower); the aryl or aralkyl optionally contains at least one heteroatom independently selected from the group consisting of: O, S, and N;
A is a divalent spacer group comprising a non-reactive divalent hydrocarbyl group having from 2 to 15 carbon atoms; Y Pg is an amino protecting group b) deprotecting the amino protecting group to give the desired resin of formula 103; c) coupling the resin with one or more amino acids in a sequential manner by standard chemistry; Y d) cutting the peptide from the resin to obtain a peptidyl-activated ketone of formula II. eleven . The process of claim 10, characterized in that the cutting step is carried out in THF, ac HCl. , and AcOH at a temperature of about 60 ° C for about 4 hours; and the resin is filtered at least once.
12. The process of claim 10, characterized in that the resin is selected from the group consisting of: polystyrene or pegylated polystyrene functionalized with benzydrylamine (BHA); 4-methyl benzidrilamine (MBHA); and aminomethyl (AM).
13. The process of claim 10, characterized in that the activation in si tu is carried out with the addition of a coupling agent selected from the group consisting of: 2- (L-T-benzotriazol-1-yl) -1,1-tetrafluoroborate. , 3, 3-tetramethyluronium (TBTU); 2- (li? -benzotriazol-1-yl) -1,3,3-tetramethyluronium hexafluorophosphate (HBTU); diisopropyl carbodiimide (DIC), and dicyclohexyl carbodiimide (DCC).
14. The process of claim 10, characterized in that the amino protecting group is selected from the group consisting of: t-butyloxycarbonyl (Boc); 9-f luorenylmet-iloxy carbonyl (Fmoc); and allyloxy carbonyl (Alloc).
15. The process of claim 10, characterized in that X 'is C (O) NH2CH2-f-enyl or C (O) OCH2CH = CH2.
16. The process of claim 10, characterized in that R is selected from the group consisting of: CH3; CH2CH3; CH2CH2CH3; ' (CH2) 4 NH2; CH (CH3) 2; CH2- phenyl; (CH2) 3-NH-CH- (NH2).
17. The process of claim 10, characterized in that A is cyclohexyl, phenyl or benzyl.
18. A resin of formula 103: 103 characterized in that R, X 'and A are as defined in claim 10.
19. The resin according to claim 17, characterized in that the resin is selected from the group consisting of: polystyrene or pegylated polystyrene functionalized with benzydrylamine (BHA); 4-methyl benzidrilamine (MBHA); and aminomethyl (AM).
20. The resin according to claim 17, characterized in that A is cyclohexyl, phenyl or benzyl.
21. The use of a resin of formula 103 for the solid phase synthesis of peptidyl-activated ketones: characterized in that R, X 'and A are as defined in claim 9.
22. The use of the resin according to claim 20, characterized in that the resin is selected from the group consisting of: polystyrene or pegylated polystyrene functionalized with benzydrylamine (BHA); 4-methyl benzidrilamine (MBHA); and aminomethyl (AM).
23. The use according to claim 20, characterized in that A is cyclohexyl, phenyl or benzyl.
MXPA/A/1999/005961A 1996-12-27 1999-06-23 Peptidomimetic inhibitors of the human cytomegalovirus protease MXPA99005961A (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US059806 1987-06-09
US034041 1996-12-27
US60/034041 1996-12-27
US052860 1997-07-17
US60/052860 1997-07-17
US60/059806 1997-09-23

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MXPA99005961A true MXPA99005961A (en) 2001-05-17

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