GLYCOPEPTIDE DERIVATIVES FOR THE PRESERVATION AND PROTECTION OF BIOLOGICAL MATERIALS AND MICROORGANISMS

Abstract

The present invention relates to a compound of the following formula (I): in which at least one and only one group chosen among R.sup.5, R.sup.6 and R.sup.7 is a group of the following formula: The present invention relates also to uses thereof for preservation and/or protection and/or regeneration of biological materials and or microorganisms and for cosmetic or dermato logical applications such as anti-aging, skin protection and skin regeneration; to culture, storage and/or preservation media comprising such a compound; to cosmetic or dermato logical compositions comprising such a compound; to processes for preparing such a compound; and to synthesis intermediates.

##STR00001##

Claims

1.-17. (canceled)

18. A compound of formula (I): ##STR00124## or a salt thereof, a solvate, a tautomer, a stereoisomer or a mixture of stereoisomers in any proportion, wherein: m is 0 or 1, p is 0 or 1, R.sub.0 is a hydrogen atom, a O-protecting group or a (C.sub.1-C.sub.6)alkyl, (C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, 5- to 7-membered heterocycloalkyl, aryl, heteroaryl, aryl-(C.sub.1-C.sub.6)alkyl, heteroaryl-(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)-alkyl-aryl or (C.sub.1-C.sub.6)-alkyl-heteroaryl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, R.sub.5a, R.sub.6a and R.sub.7a are, independently from each other, a hydrogen or a N-protecting group, and R.sub.5, R.sub.6 and R.sub.7 are, independently from each other, a hydrogen; a (C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy or a (C.sub.1-C.sub.6)thioalkoxy; an aryl; an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy; or a group of the following formula: ##STR00125## or R.sub.5 and R.sub.5a and/or R.sub.6 and R.sub.6a and/or R.sub.7 and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00126## provided that at least one and only one group among R.sub.5, R.sub.6 and R.sub.7 when m=p=1, or at least one and only one group among R.sub.5 and R.sub.6 when m=0 and p=1, or at least one and only one group among R.sub.8 and R.sub.7 when m=1 and p=0, or R.sub.5 when m=p=0, is a group of formula: ##STR00127## with n is an integer from 1 to 6, R is a hydrogen or fluorine atom or a CH.sub.3, CH.sub.2F, CH.sub.2OSiR.sup.a1R.sup.b1R.sup.c1, CH.sub.2OR.sub.8, CH.sub.2OC(O)R.sub.9, CH.sub.2OCO.sub.2R.sub.10, CH.sub.2OC(O)NR.sub.11R.sub.12, CH.sub.2OP(O)(OR.sub.13).sub.2 or CH.sub.2OSO.sub.3R.sub.14 group, R.sub.1 and R.sub.2 are, independently from one another, a fluorine atom or an OSiR.sup.a2R.sup.b2R.sup.c2, OR.sub.15, OC(O)R.sub.16, OCO.sub.2R.sub.17, OC(O)NR.sub.18R.sub.19, OP(O)(OR.sub.20).sub.2 or OSO.sub.3R.sub.21 group, R.sub.3 are a fluorine atom or an OSiR.sup.a3R.sup.b3R.sup.c3, OR.sub.22, OC(O)R.sub.23, OCO.sub.2R.sub.24, OCONR.sub.25R.sub.26, OP(O)(OR.sub.27).sub.2, OSO.sub.3R.sub.28, N.sub.3, phtalimidyl, NR.sub.29R.sub.30, NR.sub.31C(O)R.sub.32, NR.sub.33C(O)OR.sub.34, N(C(O)R.sub.35)C(O)R.sub.36, N(C(O)R.sub.37)C(O)OR.sub.38 and N(C(O)OR.sub.39)C(O)OR.sub.40 group, R.sub.4 is a hydrogen or halogen atom or an OSiR.sup.a4R.sup.b4R.sup.c4, OR.sub.41, OC(O)R.sub.42, OCO.sub.2R.sub.43, OCONR.sub.44R.sub.45, OP(O)(OR.sub.46).sub.2, or OSO.sub.3R.sub.47 group, or R and R.sub.1, together with the carbon atoms carrying them, form a cyclic acetal having the following formula: ##STR00128## and/or (R.sub.1 and R.sub.2), (R.sub.2 and R.sub.3), and/or (R.sub.3 and R.sub.4), together with the carbon atoms carrying them, form a cyclic acetal having the following formula: ##STR00129## R.sub.8, R.sub.15, R.sub.22 and R.sub.41 are, independently from one another, a hydrogen atom, a O-protecting group or a (C.sub.1-C.sub.6)alkyl, (C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, 5- to 7-membered heterocycloalkyl, aryl, heteroaryl, aryl-(C.sub.1-C.sub.6)alkyl, heteroaryl-(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)-alkyl-aryl, (C.sub.1-C.sub.6)-alkyl-heteroaryl, saccharidic or polysaccharidic group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, R.sub.9, R.sub.10, R.sub.16, R.sub.17, R.sub.23, R.sub.24, R.sub.32, R.sub.34 to R.sub.40, R.sub.42 and R.sub.43 are, independently from one another, a (C.sub.1-C.sub.6)alkyl, (C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, 5- to 7-membered heterocycloalkyl, aryl, heteroaryl, aryl-(C.sub.1-C.sub.6)alkyl, heteroaryl-(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)-alkyl-aryl or (C.sub.1-C.sub.6)-alkyl-heteroaryl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, R.sub.11, R.sub.12, R.sub.18, R.sub.19, R.sub.25, R.sub.26, R.sub.29 to R.sub.31, R.sub.33, R.sub.44 and R.sub.45 are, independently from one another, a hydrogen atom or a (C.sub.1-C.sub.6)alkyl, (C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, aryl, heteroaryl, aryl-(C.sub.1-C.sub.6)alkyl, heteroaryl-(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)-alkyl-aryl or (C.sub.1-C.sub.6)-alkyl-heteroaryl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, R.sub.13, R.sub.14, R.sub.20, R.sub.21, R.sub.27, R.sub.28, R.sub.46 and R.sub.47 are, independently from one another, a hydrogen atom or a (C.sub.1-C.sub.6)alkyl group, R.sup.a1 to R.sup.a4, R.sup.b1 to R.sup.b4 and R.sup.c1 to R.sup.c4 are, independently from one another, a (C.sub.1-C.sub.6)alkyl, aryl or aryl-(C.sub.1-C.sub.6)alkyl group, and R.sup.d and R.sup.e are, independently from one another, a hydrogen atom or a (C.sub.1-C.sub.6)alkyl group.

19. The compound according to claim 18, wherein it is a compound of the following formula (Ie1), (Ii1), (Im1) or (Iq1): ##STR00130## or a salt, a solvate or a tautomer thereof.

20. The compound according to claim 18, wherein R.sub.5, R.sub.6 and R.sub.7 are, independently from each other, a hydrogen atom; a (C.sub.1-C.sub.6)alkyl; an aryl; an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy group, or a group of the following formula: ##STR00131## or R.sub.5 and R.sub.5a and/or R.sub.6 and R.sub.6a and/or R.sub.7 and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00132## provided that at least one and only one group among R.sub.5, R.sub.6 and R.sub.7 when m=p=1, or at least one and only one group among R.sub.5 and R.sub.6 when m=0 and p=1, or at least one and only one group among R.sub.5 and R.sub.7 when m=1 and p=0, or R.sub.5 when m=p=0, is a group of formula: ##STR00133##

21. The compound according to claim 18, wherein R.sub.5, R.sub.6 and R.sub.7 are, independently from each other, H, CH.sub.3, CH(CH.sub.3).sub.2, CH.sub.2CH(CH.sub.3).sub.2, CH(CH.sub.3)CH.sub.2CH.sub.3, CH.sub.2Ph, CH.sub.2PhOCH.sub.3, CH.sub.2CH.sub.2SCH.sub.3 or a group of the following formula: ##STR00134## or R.sub.5 and R.sub.5a and/or R.sub.6 and R.sub.6a and/or R.sub.7 and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00135## provided that at least one and only one group among R.sub.5, R.sub.6 and R.sub.7 when m=p=1, or at least one and only one group among R.sub.5 and R.sub.6 when m=0 and p=1, or at least one and only one group among R.sub.5 and R.sub.7 when m=1 ad p=0, or R.sub.5 when m=p=0, is a group of formula: ##STR00136##

22. The compound according to claim 18, wherein n has the value 2, 3 or 4.

23. The compound according to claim 18, wherein R.sub.0 is a hydrogen atom or a (C.sub.1-C.sub.6)alkyl, aryl or aryl-(C.sub.1-C.sub.6)alkyl group, this group being unsubstituted or substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO; R is a CH.sub.2OR.sub.8 group; R.sub.1 and R.sub.2 are, independently from one another, an OR.sub.15 group; and R.sub.3 is an OR.sub.22 group, with R.sub.8, R.sub.15 and R.sub.22 are a hydrogen atom or an O-protecting group; and R.sub.4 is a hydrogen atom or an OR.sub.41 group, wherein R.sub.41 is a hydrogen atom, a O-protecting group or a (C.sub.1-C.sub.6)alkyl, aryl, aryl-(C.sub.1-C.sub.6)alkyl, or (C.sub.1-C.sub.6)-alkyl-aryl group, this group being unsubstituted or possibly substituted with one or more groups selected from a halogen atom and (C.sub.1-C.sub.6)alkoxy.

24. The compound according to claim 18, wherein R.sub.0H, Et or Bn, RCH.sub.2OH or CH.sub.2OBn, R.sub.1R.sub.2R.sub.3OH or OBn and R.sub.4H, OH, OMe or OBn.

25. The compound according to claim 18, wherein R.sub.0H, RCH.sub.2OH, R.sub.1R.sub.2R.sub.3OH and R.sub.4H or OH.

26. The compound according to claim 18, wherein it is selected from the following compounds: ##STR00137## ##STR00138## ##STR00139## ##STR00140## ##STR00141##

27. A culture, storage and/or preservation medium comprising at least one compound according to claim 18.

28. A cosmetic or dermatological composition comprising at least one compound according to claim 18 and at least one cosmetically or dermatologically acceptable excipient.

29. A process for preparing a compound of formula (I) according to claim 18 comprising the following successive steps: (a) reducing the imine function of a compound of the following formula (II): ##STR00142## wherein: R.sub.5b, R.sub.6b and R.sub.7b are, independently from each other, a hydrogen; a (C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy or a (C.sub.1-C.sub.6)thioalkoxy; an aryl; an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy; or a group of the following formula: ##STR00143## or R.sub.5b and R.sub.5a and/or R.sub.6b and R.sub.6a and/or R.sub.7b and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00144## provided that at least one and only one group among R.sub.5b, R.sub.6b and R.sub.7b when m=p=1, or at least one and only one group among R.sub.5b and R.sub.6b when m=0 and p=1, or at least one and only one group among R.sub.5b and R.sub.7b when m=1 and p=0, or R.sub.5b when m=p=0, is a group of formula: ##STR00145## to obtain a compound of formula (I), and (b) optionally salifying or solvating the compound obtained in previous step (a) to obtain a salt or solvate of a compound of formula (I).

30. A process for preparing a compound of formula (I) according to claim 18 comprising the following successive steps: (i) reacting a compound of the following formula (IX): ##STR00146## wherein LG is a leaving group, with a compound of the following formula (III): ##STR00147## or a salt thereof, wherein: R.sub.5c, R.sub.6c and R.sub.7c are, independently from each other, a hydrogen; a (C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy or a (C.sub.1-C.sub.6)thioalkoxy; an aryl; an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy; or a group of the following formula: ##STR00148## or R.sub.5c and R.sub.5a and/or R.sub.6c and R.sub.6a and/or R.sub.7c and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00149## provided that at least one and only one group among R.sub.5c, R.sub.6c and R.sub.7c when m=p=1, or at least one and only one group among R.sub.5c and R.sub.6c when m=0 and p=1, or at least one and only one group among R.sub.5c and R.sub.7c when m=1 and p=0, or R.sub.5c when m=p=0, is a group of formula: ##STR00150## to obtain a compound of formula (I), and (ii) optionally salifying or solvating the compound obtained in previous step (i) to obtain a salt or solvate of a compound of formula (I).

31. A process for preparing a compound of formula (I) according to claim 18 where m and p are not both 0 and R.sub.5 is a group of the following formula: ##STR00151## comprising the following successive steps: (1) reacting a compound of the following formula (XIa): ##STR00152## or a salt thereof, with a compound of the following formula (VII) ##STR00153## or a salt thereof, provided that m and p are not both 0, and R.sub.6d and R.sub.7d are, independently of each other, a hydrogen; a (C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy or a (C.sub.1-C.sub.6)thioalkoxy; an aryl; or an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy; or R.sub.6d and R.sub.6a and/or R.sub.7d and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00154## to obtain a compound of formula (I), and (2) optionally salifying or solvating the compound obtained in previous step (1) to obtain a salt or solvate of a compound of formula (I).

32. A compound of formula (II), (VI) or (XI): ##STR00155## or a salt thereof, a solvate, a tautomer, a stereoisomer or a mixture of stereoisomers in any proportion, wherein: n is an integer from 1 to 6, m is 0 or 1, p is 0 or 1, R.sub.0 is a hydrogen atom, a O-protecting group or a (C.sub.1-C.sub.6)alkyl, (C.sub.2-C.sub.6)alkenyl, (C.sub.2-C.sub.6)alkynyl, (C.sub.3-C.sub.7)cycloalkyl, 5- to 7-membered heterocycloalkyl, aryl, heteroaryl, aryl-(C.sub.1-C.sub.6)alkyl, heteroaryl-(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)-alkyl-aryl or (C.sub.1-C.sub.6)-alkyl-heteroaryl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, R is a hydrogen or fluorine atom or a CH.sub.3, CH.sub.2F, CH.sub.2OSiR.sup.a1R.sup.b1R.sup.c1, CH.sub.2OR.sub.8, CH.sub.2OC(O)R.sub.9, CH.sub.2OCO.sub.2R.sub.10, CH.sub.2OC(O)NR.sub.11R.sub.12, CH.sub.2OP(O)(OR.sub.13).sub.2 or CH.sub.2OSO.sub.3R.sub.14 group, R.sub.1 and R.sub.2 are, independently from one another, a fluorine atom or an OSiR.sup.a2R.sup.b2R.sup.c2, OR.sub.15, OC(O)R.sub.16, OCO.sub.2R.sub.17, OC(O)NR.sub.18R.sub.19, OP(O)(OR.sub.20).sub.2 or OSO.sub.3R.sub.21 group, R.sub.3 are a fluorine atom or an OSiR.sup.a3R.sup.b3R.sup.c3, OR.sub.22, OC(O)R.sub.23, OCO.sub.2R.sub.24, OCONR.sub.25R.sub.26, OP(O)(OR.sub.27).sub.2, OSO.sub.3R.sub.28, N.sub.3, phtalimidyl, NR.sub.29R.sub.30, NR.sub.31C(O)R.sub.32, NR.sub.33C(O)OR.sub.34, N(C(O)R.sub.35)C(O)R.sub.36, N(C(O)R.sub.37)C(O)OR.sub.38 and N(C(O)OR.sub.39)C(O)OR.sub.40 group, R.sub.4 is a hydrogen or halogen atom or an OSiR.sup.a4R.sup.b4R.sup.c4, OR.sub.41, OC(O)R.sub.42, OCO.sub.2R.sub.43, OCONR.sub.44R.sub.45, OP(O)(OR.sub.46).sub.2, or OSO.sub.3R.sub.47 group, or R and R.sub.1, together with the carbon atoms carrying them, form a cyclic acetal having the following formula: ##STR00156## and/or (R.sub.1 and R.sub.2), (R.sub.2 and R.sub.3), and/or (R.sub.3 and R.sub.4), together with the carbon atoms carrying them, form a cyclic acetal having the following formula: ##STR00157## R.sub.5a, R.sub.6a and R.sub.7a are, independently from each other, a hydrogen or a N-protecting group, R.sub.5b, R.sub.6b and R.sub.7b are, independently from each other, a hydrogen; a (C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy or a (C.sub.1-C.sub.6)thioalkoxy; an aryl; an aryl-(C.sub.1-C.sub.6)alkyl optionally substituted by a (C.sub.1-C.sub.6)alkoxy; or a group of the following formula: ##STR00158## or R.sub.5b and R.sub.5a and/or R.sub.6b and R.sub.6a and/or R.sub.7b and R.sub.7a form, with the nitrogen and carbon atoms carrying them, a pyrrolidine cycle ##STR00159## provided that at least one and only one group among R.sub.5b, R.sub.6b and R.sub.7b when m=p=1, or at least one and only one group among R.sub.5b and R.sub.6b when m=0 and p=1, or at least one and only one group among R.sub.5b and R.sub.7b when m=1 and p=0, or R.sub.5b, when m=p=0, is a group of formula: ##STR00160## R.sub.49 is H or a O-protecting group, and PG is a O-protecting group.

33. The compound according to claim 18, wherein: R.sub.8, R.sub.11, R.sub.12, R.sub.15, R.sub.18, R.sub.19, R.sub.22, R.sub.25, R.sub.26, R.sub.29 to R.sub.31, R.sub.33, R.sub.41, R.sub.44 and R.sub.45 are a hydrogen atom or a (C.sub.1-C.sub.6)alkyl, aryl or aryl-(C.sub.1-C.sub.6)alkyl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C.sub.1-C.sub.6)alkoxy, OH, COOH and CHO, and R.sub.9, R.sub.10, R.sub.16, R.sub.17, R.sub.23, R.sub.24, R.sub.32, R.sub.34 to R.sub.40, R.sub.42 and R.sub.43 are a (C.sub.1-C.sub.6)alkyl, aryl or aryl-(C.sub.1-C.sub.6)alkyl group, this group being possibly substituted with one or more groups selected from a halogen atom, (C1-C6)alkoxy, OH, COOH and CHO.

34. The compound according to claim 21 wherein ##STR00161## has the following configuration: ##STR00162##

35. A method of preservation and/or protection of a biological material or of a microorganism comprising placing said biological material or microorganism in a medium containing a compound according to claim 18.

36. The method according to claim 35, wherein the biological material is selected from cells, tissues, body fluids and organs.

37. A method for anti-aging, skin protection or skin regeneration comprising applying to the skin of a person in need thereof of an affective amount of a compound according to claim 18.

Description

FIGURES

[0295] FIG. 1 represents the percentage of fibroblasts viability during time (from 0 to 12 days) after incubation in a common culture medium (surviving control), in a starvation medium (serum-free control) and in a starvation medium containing compound 17 at 5 mg/mL.

[0296] FIGS. 2 and 3 represent respectively the .sup.1H RMN spectrum of a solution of compound 17 after addition of 0 or 8 equivalents of NaOD and 2 h 30 of reaction.

[0297] FIGS. 4 and 5 represent respectively the .sup.1H RMN spectrum of a solution of compound X after addition of 0 or 8 equivalents of NaOD and 2 h 30 of reaction.

[0298] FIG. 6 represents the .sup.1H RMN spectrum of a solution of compound X after addition of 8 equivalents of NaOD and 3 days of reaction.

[0299] FIGS. 7 and 8 represent the mass spectra of a solution of compound X after addition of 8 equivalents of NaOD and 3 days of reaction, respectively in ESI.sup.+ and ESI.sup. mode.

EXAMPLES

[0300] The following abbreviations have been used: [0301] Ala: Alanine [0302] Ac: Acetyl (COCH.sub.3) [0303] Bn: Benzyl (CH.sub.2Ph) [0304] Cbz: Benzyloxycarbonyl (CO.sub.2CH.sub.2Ph) [0305] CDI: Carbonyldiimidazole [0306] Dab: 2,4-Diaminobutyric acid [0307] DCE: Dichloroethane [0308] DMEM: Dulbecco's Modified Eagle Medium [0309] DMF: Dimethylformamide [0310] DIBAl-H: Diisobutylaluminium hydride [0311] DIEA: N,N-Diisopropylethylamine [0312] EDTA: Ethylenediaminetetraacetic acid [0313] ESI: Electrospray ionisation [0314] Et: Ethyl (CH.sub.2CH.sub.3) [0315] FBS: Fetal Bovine Serum [0316] Gly: Glycine [0317] Lys: Lysine [0318] Me: Methyl (CH.sub.3) [0319] MEM: Minimum Essential Media [0320] NMM: N-Methylmorpholine [0321] NMR: Nuclear Magnetic Resonance [0322] OD: Optical density [0323] Orn: Ornithine [0324] PBS: Phosphate buffered saline [0325] Pro: Proline [0326] PSNEt.sub.2: Diethylaminomethyl-polystyrene [0327] PyBOP: (1H-Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate [0328] Tf: Trifluoromethanesulfonyl (SO.sub.2CF.sub.3) [0329] TFA: Trifluoroacetic acid [0330] THF: Tetrahydrofuran [0331] Tyr: Tyrosine [0332] Z: Benzyloxycarbonyl (CO.sub.2CH.sub.2Ph)

ISynthesis of the Compounds According to the Invention

[0333] It should be noted that the compounds according to the invention where R.sub.4R.sub.1OH can be obtained in the form of a mixture of tautomer forms as explained in the description above. For practical reasons, these compounds are represented by their pyranose form. It concerns compounds 17, 19, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40 and 44 according to the invention.

I-1. General procedure

Synthesis of Peptide Intermediates

[0334] ##STR00099## [0335] (where aa.sub.1 and aa.sub.2 each represent independently a residue of amino acid)

Example of Synthesis of Peptide Intermediates

General Procedure A

[0336] Boc-aa.sub.1-OH and HCl.H.sub.2N-aa.sub.2-OBn coupling: Boc-aa.sub.1-OH was dissolved in dichloromethane and carbonyldiimidazole (1.03 eq.) was added portionwise. After stirring during 1 h, a solution of HCl.H.sub.2N-aa.sub.2-OBn (1 eq.) and DIEA (2.1 eq.) in dichloromethane was added dropwise. The reaction mixture was stirred at room temperature overnight. Aqueous HCl 1N was added and the mixture was vigorously stirred for 10 min. The layers were separated and the aqueous one was extracted with dichloromethane. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The obtained residue was purified by flash column chromatography to give Boc-aa.sub.1-aa.sub.2-OBn.

General Procedure B

[0337] N-deprotection using TFA: N-Boc peptide was dissolved in dichloromethane and trifluoroacetic acid (20 eq.) was added dropwise. The reaction mixture was stirred at room temperature overnight before being concentrated under vacuum to give ammonium trifluoroacetate salt of the peptide (X.sup.CF.sub.3CO.sub.2.sup.).

General Procedure C

[0338] N-deprotection using HCl: N-Boc peptide was dissolved in dichloromethane and a 4M solution of HCl in dioxane (10 eq.) was added dropwise. The reaction mixture was stirred at room temperature for 2 h before being concentrated under vacuum to give ammonium hydrochloride salt of the peptide (X.sup.Cl.sup.).

General Procedure D

[0339] Z,Boc-protected amino acid

##STR00100##

and .sup.X..sup.+H.sub.3N-aa.sub.1-aa.sub.2-OBn coupling: Z,Boc-protected amino acid was dissolved in dichloromethane and carbonyldiimidazole (1.2 eq.) was added portionwise. After stirring during 1 h, a solution of .sup.X..sup.+H.sub.3N-aa.sub.1-aa.sub.2-OBn (1 eq.) and DIEA (2.1 eq.) in dichloromethane was added dropwise. The reaction mixture was stirred at room temperature overnight. An aqueous solution of 1M HCl was added and the mixture was vigorously stirred for 10 min. The layers were separated and the aqueous one was extracted with dichloromethane. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The obtained residue was purified by flash column chromatography to give the coupling residue.

[0340] Boc-Ala-Ala-OBn:

[0341] Using general procedure A with Boc-Ala-OH (10 g, 52.9 mmol, 1 eq.) and HCl.H.sub.2N-Ala-OBn (11.4 g, 52.9 mmol, 1 eq.). Pale yellow oil (18.2 g, 98%). MS (ESI.sup.+): 351.2 [M+H].sup.+; 373.2 [M+Na].sup.+; 389.1 [M+K].sup.+

[0342] HCl.H.sub.2N-Ala-Ala-OBn:

[0343] Using general procedure C starting from Boc-Ala-Ala-OBn (18.04 g, 51.5 mmol, 1 eq.). White solid (14.75 g, 100%).

[0344] MS (ESI.sup.+): 251.1 [M-HCl+H].sup.+; 273.1 [M-HCl+Na].sup.+; 289.1 [M-HCl+K].sup.+

[0345] TFA.H.sub.2N-Ala-Ala-OBn:

[0346] Using general procedure B starting from Boc-Ala-Ala-OBn (17.2 g, 49.03 mmol, 1 eq.). Pale yellow oil (17.9 g, 100%).

[0347] .sup.1H NMR (CDCl.sub.3, 300 MHz): 1.3-1.5 (m, 6H), 4.0-4.2 (m, 1H), 4.3-4.5 (m, 1H), 5.02 (d, 1H, .sup.1J.sub.H-H=11.5 Hz), 5.11 (d, 1H, .sup.1J.sub.H-H=11.5 Hz), 7.1-7.4 (m, 5H), 7.65 (d, 1H, .sup.3J.sub.H-H=6 Hz), 7.8-8.3 (m, 2H)

[0348] Z-Lys(Boc)-Ala-Ala-OBn:

[0349] Using general procedure D starting from HCl.H.sub.2N-ALa-Ala-OBn (15 g, 52.3 mmol, 1 eq.) and Z-Lys(Boc)-OH (19.9 g, 52.5 mmol, 1 eq.). White solid (21.2 g, 67%).

[0350] MS (ESI.sup.+): 613.3 [M+H].sup.+; 635.2 [M+Na].sup.+; 651.2 [M+K].sup.+

[0351] Z-Lys(HCl)-Ala-Ala-OBn:

[0352] Using general procedure C starting from Z-Lys(Boc)-Ala-Ala-OBn (21 g, 34.3 mmol, 1 eq.). White solid (16.4 g, 87%).

[0353] MS (ESI.sup.+): 513.2 [M-HCl+H].sup.+; 535.2 [M-HCl+Na].sup.+; 551.2 [M-HCl+K].sup.+

[0354] Z-Lys(Boc)-Ala-OBn:

[0355] Using general procedure D with Z-Lys(Boc)-OH (2 g, 5.26 mmol, 1 eq.) and HCl.H.sub.2N-Ala-OBn (1.13 g, 5.26 mmol, 1 eq.) instead of HCl.H.sub.2N-aa.sub.1-aa.sub.2-OBn. Pale yellow solid (1.89 g, 66%).

[0356] .sup.1H NMR_(CDCl.sub.3, 300 MHz): 1.3 (m, 16H); 1.6-1.7 (m, 2H); 3.0 (m, 2H); 4.1 (m, 1H); 4.5 (m, 1H); 4.6 (m, 1H); 5.0 (s, 2H); 5.1 (m, 1H); 5.5 (d, 7 Hz, 1H); 6.8 (d, 6.5 Hz, 1H); 7.3 (m, 10H).

[0357] Z-Lys(HCl)-Ala-OBn:

[0358] Using general procedure C starting from Z-Lys(Boc)-Ala-OBn (756 mg, 1.40 mmol, 1 eq.). White solid (778 mg, 100%).

[0359] MS (ESI.sup.+): 223.1 [M+H].sup.+; 245.1 [M+Na].sup.+; 261.1 [M+K].sup.+

[0360] Boc-Gly-Gly-OBn:

[0361] Using general procedure A with Boc-Gly-OH (800 mg, 4.57 mmol, 1 eq.) and HCl.H.sub.2N-Gly-OBn (922 mg, 4.57 mmol, 1 eq.). Colorless oil (1.40 g, 95%).

[0362] MS (ESI.sup.+): 323.2 [M+H].sup.+; 345.1 [M+Na].sup.+; 361.1 [M+K].sup.+

[0363] HCl.H.sub.2N-Gly-Gly-OBn:

[0364] Using general procedure C starting from Boc-Gly-Gly-OBn (1.37 g, 4.25 mmol, 1 eq.). White solid (1.14 g, 100%).

[0365] MS (ESI.sup.+): 223.1[M-HCl+H].sup.+; 245.1 [M-HCl+Na].sup.+; 261.1 [M-HCl+K].sup.+

[0366] Z-Lys(Boc)-Gly-Gly-OBn:

[0367] Using general procedure D starting from HCl.H.sub.2N-Gly-Gly-OBn (1.12 g, 4.17 mmol, 1 eq.) and Z-Lys(Boc)-OH (1.68 g, 4.17 mmol, 1 eq.). Yellow oil (1.69 g, 69%).

[0368] MS (ESI.sup.+): 585.3 [M+H].sup.+; 607.2 [M+Na].sup.+; 623.2 [M+K].sup.+

[0369] Z-Lys(HCl)-Gly-Gly-OBn:

[0370] Using general procedure C starting from Z-Lys(Boc)-Gly-Gly-OBn (1.60 g, 2.79 mmol, 1 eq.). White solid (1.59 g, 100%).

[0371] MS (ESI.sup.+): 485.2 [M-HCl+H].sup.+; 507.2 [M-HCl+Na].sup.+; 523.2 [M-HCl+K].sup.+

[0372] Boc-Ala-Gly-OBn:

[0373] Using general procedure A with Boc-Ala-OH (400 mg, 2.1 mmol, 1 eq.) and HCl.H.sub.2N-Gly-OBn (425 mg, 2.1 mmol, 1 eq.). Colorless oil (666 mg, 94%).

[0374] MS (ESI.sup.+): 359.2 [M+Na].sup.+; 375.1 [M+K].sup.+

[0375] TFA.H.sub.2N-Ala-Gly-OBn:

[0376] Using general procedure B starting from Boc-Ala-Gly-OBn (646 mg, 1.92 mmol, 1 eq.). Yellow oil (759 mg, 100%).

[0377] MS (ESI.sup.+): 237.1 [M-TFA+H].sup.+; 259.1 [M-TFA+Na].sup.+

[0378] Z-Lys(Boc)-Ala-Gly-OBn:

[0379] Using general procedure D starting from TFA.H.sub.2N-Ala-Gly-OBn (739 mg, 1.87 mmol, 1 eq.) and Z-Lys(Boc)-OH (711 mg, 1.87 mmol, 1 eq.). White solid (797 mg, 71%).

[0380] MS (ESI.sup.+): 621.4 [M+Na].sup.+; 637.4 [M+K].sup.+

[0381] Z-Lys(TFA)-Ala-Gly-OBn:

[0382] Using general procedure B starting from Z-Lys(Boc)-Ala-Gly-OBn (777 mg, 1.30 mmol, 1 eq.). Yellow oil (947 mg, 100%).

[0383] MS (ESI.sup.+): 499.3 [M-TFA+H].sup.+; 521.3 [M-TFA+Na].sup.+; 537.3 [M-TFA+K].sup.+

[0384] Boc-Gly-Ala-OBn:

[0385] Using general procedure A with Boc-Gly-OH (400 mg, 2.29 mmol, 1 eq.) and HCl.H.sub.2N-Ala-OBn (494 mg, 2.29 mmol, 1 eq.). Colorless oil (703 mg, 91%).

[0386] MS (ESI.sup.+): 359.2 [M+Na].sup.+; 375.1 [M+K].sup.+

[0387] TFA.H.sub.2N-Gly-Ala-OBn:

[0388] Using general procedure B starting from Boc-Gly-Ala-OBn (683 mg, 2.03 mmol, 1 eq.). Yellow oil (772 mg, 100%).

[0389] MS (ESI.sup.+): 237.1 [M-TFA+H].sup.+; 259.1 [M-TFA+Na].sup.+

[0390] Z-Lys(Boc)-Gly-Ala-OBn:

[0391] Using general procedure D starting from TFA.H.sub.2N-Gly-Ala-OBn (753 mg, 1.98 mmol, 1 eq.) and Z-Lys(Boc)-OH (753 mg, 1.98 mmol, 1 eq.). Yellow oil (939 mg, 79%).

[0392] MS (ESI.sup.+): 621.4 [M+Na].sup.+; 637.4 [M+K].sup.+

[0393] Z-Lys(TFA)-Gly-Ala-OBn:

[0394] Using general procedure B starting from Z-Lys(Boc)-Gly-Ala-OBn (919 mg, 1.53 mmol, 1 eq.). Yellow oil (1.10 g, 100%).

[0395] MS (ESI.sup.+): 499.3 [M-TFA+H].sup.+; 521.3 [M-TFA+Na].sup.+

[0396] Z-Om(Boc)-Ala-Ala-OBn:

[0397] Using general procedure D starting from TFA.H.sub.2N-Ala-Ala-OBn (878 mg, 2.41 mmol, 1.04 eq.) and Z-Om(Boc)-OH (850 mg, 2.32 mmol, 1 eq.). White solid (879 mg, 63%).

[0398] MS (ESI.sup.+): 599.3 [M+H].sup.+; 621.3 [M+Na].sup.+; 637.3 [M+K].sup.+

[0399] Z-Omrn(HCl)-Ala-Ala-OBn:

[0400] Using general procedure C starting from Z-Om(Boc)-Ala-Ala-OBn (874 mg, 1.46 mmol, 1 eq.). White solid (822 mg, 100%).

[0401] MS (ESI.sup.+): 499.3 [M-HCl+H].sup.+; 521.2 [M-HCl+Na].sup.+

[0402] Z-Dab(Boc)-Ala-Ala-OBn:

[0403] Using general procedure D starting from TFA.H.sub.2N-Ala-Ala-OBn (463 mg, 1.27 mmol, 1 eq.) and Z-Dab(Boc)-OH (447 mg, 1.27 mmol, 1 eq.). Z-Dab(Boc)-OH was prepared from Z-Dab(Boc)-OH.DCHA (dicyclohexylamine salt) as follows: The salt was diluted in dichloromethane, this solution was washed three times with a saturated aqueous solution of potassium hydrogenosulfate, dried over anhydrous sodium sulfate and concentrated under vacuum. White solid (402 mg, 54%).

[0404] MS (ESI.sup.+): 585.3 [M+H].sup.+; 607.3 [M+Na].sup.+; 623.3 [M+K].sup.+

[0405] Z-Dab(HCl)-Ala-Ala-OBn:

[0406] Using general procedure C starting from Z-Dab(Boc)-Ala-Ala-OBn (400 mg, 0.684 mmol, 1 eq.). White solid (413 mg, 100%).

[0407] MS (ESI.sup.+): 485.2 [M-HCl+H].sup.+; 507.2 [M-HCl+Na].sup.+

[0408] Boc-Ala-Pro-OBn:

[0409] Boc-Ala-OH (1.00 g, 5.28 mmol, 1 eq.) was dissolved in dichloromethane (8 mL) under N.sub.2 atmosphere. HCl.H.sub.2N-Pro-OBn (1.40 g, 5.81 mmol, 1.1 eq.) and 2-bromoethylpyridinium (1.59 g, 5.81 mmol, 1.1 eq.) were added sequentially. The reaction mixture was cooled to 0 C. and N,N-diisopropylethylamine (2.8 mL, 16.9 mmol, 3.2 eq.) was added dropwise. The mixture was allowed to warm to room temperature and stirred for 5 h. After completion of the reaction, the reaction mixture was diluted with dichloromethane (20 mL). The obtained solution was washed with an aqueous solution of citric acid (10%, 15 mL), then with an aqueous solution of sodium hydrogenocarbonate (5%, 15 mL) before being dried over anhydrous sodium sulfate and concentrated under vacuum. The obtained residue was purified by flash column chromatography to give Boc-Ala-Pro-OBn as a pale yellow oil (1.34 g, 67%).

[0410] MS (ESI.sup.+): 399.2 [M+Na].sup.+

[0411] TFA.H.sub.2N-Ala-Pro-OBn:

[0412] Using general procedure B, starting from Boc-Ala-Pro-OBn (1.27 g, 3.7 mmol, 1 eq.). Pale brown oil (1.31 g, 100%).

[0413] MS (ESI.sup.+): 277.2 [M-TFA+H].sup.+; 299.1 [M-TFA+Na].sup.+

[0414] Z-Lys(Boc)-Ala-Pro-OBn:

[0415] Using general procedure D starting from Z-Lys(Boc)-OH (1.30 mg, 3.43 mmol, 1 eq.) and TFA.H.sub.2N-Ala-Pro-OBn (1.34 mg, 3.43 mmol, 1 eq.). White solid (1.24 g, 57%).

[0416] MS (ESI.sup.+): 639.3 [M+H].sup.+; 661.3 [M+Na].sup.+; 677.3 [M+K].sup.+

[0417] Z-Lys(TFA)-Ala-Pro-OBn:

[0418] Using general procedure B, starting from Z-Lys(Boc)-Ala-Pro-OBn (1.08 g, 1.69 mmol, 1 eq.). Pale brown solid (1.1 g, 56%).

[0419] MS (ESI.sup.+): 539.3 [M-TFA+H].sup.+

[0420] Boc-Pro-Ala-OBn:

[0421] Using general procedure A with Boc-Pro-OH (2.0 g, 9.29 mmol, 1 eq.) and HCl.H.sub.2N-Ala-OBn (2.04 mg, 9.29 mmol, 1 eq.). White solid (2.79 g, 80%).

[0422] MS (ESI.sup.+): 399.2 [M+Na].sup.+; 415.2 [M+K].sup.+

[0423] TFA.H.sub.2N-Pro-Ala-OBn:

[0424] Using general procedure B starting from Boc-Pro-Ala-OBn (537 mg, 1.43 mmol, 1 eq.). Pale yellow solid (558 mg, 100%).

[0425] MS (ESI.sup.+): 277.2 [M-TFA+H].sup.+; 299.1 [M-TFA+Na].sup.+

[0426] Z-Lys(Boc)-Pro-Ala-OBn:

[0427] TFA.H.sub.2N-Pro-Ala-OBn (542 mg, 1.39 mmol, 1.1 eq.) was dissolved in dichloromethane (7 mL) under N.sub.2 atmosphere. Z-Lys(Boc)-OH (479 mg, 1.26 mmol, 1 eq.) and 2-bromoethylpyridinium (380 mg, 1.39 mmol, 1.1 eq.) were added sequentially. The reaction mixture was cooled to 0 C. and N,N-diisopropylethylamine (670 L, 4.03 mmol, 3.2 eq.) was added dropwise. The mixture was allowed to warm to room temperature and stirred for 5 h. After completion of the reaction, the reaction mixture was diluted with dichloromethane (15 mL). The obtained solution was washed with an aqueous solution of citric acid (10%, 10 mL), then with an aqueous solution of sodium hydrogenocarbonate (5%, 10 mL) before being dried over anhydrous sodium sulfate and concentrated under vacuum. The obtained residue was purified by flash column chromatography to give Z-Lys(Boc)-Pro-Ala-OBn as a white solid (382 mg, 43%).

[0428] MS (ESI.sup.+): 639.5 [M+H].sup.+; 661.5 [M+Na].sup.+; 677.4 [M+K].sup.+

[0429] Z-Lys(TFA)-Pro-Ala-OBn:

[0430] Using general procedure B starting from Z-Lys(Boc)-Pro-Ala-OBn (358 mg, 0.56 mmol, 1 eq.). Pale brown solid (365 mg, 100%).

[0431] MS (ESI.sup.+): 539.3 [M-TFA+H].sup.+

[0432] Z-Ala-Lys(Boc)-OMe:

[0433] Using general procedure A starting from Z-Ala-OH (500 mg, 2.24 mmol, 1 eq.) and HCl.H.sub.2N-Lys(Boc)-OMe (665 mg, 2.24 mmol, 1 eq.). Viscous colorless oil (863 mg, 83%).

[0434] MS (ESI.sup.+): 466.3 [M+H].sup.+; 488.2 [M+Na].sup.+; 504.2 [M+K].sup.+

[0435] Z-Ala-Lys(Boc)-OH:

[0436] To a solution of Z-Ala-Lys(Boc)-OMe (786 mg, 1.69 mmol) in tetrahydrofuran (49 mL) was added a solution of lithium hydroxide (121 mg, 3 eq.) in water (5.6 mL). The reaction mixture was stirred for 16 h then a solution of aqueous hydrochloric acid (1M) was added until pH 1 and the mixture was extracted 3 times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum to give a pale yellow oil (763 mg, 100%).

[0437] MS (ESI.sup.+): 452.2 [M+H].sup.+; 474.2 [M+Na].sup.+; 490.2 [M+K].sup.+

[0438] Z-Ala-Lys(Boc)-Ala-OBn:

[0439] Using general procedure A starting from Z-Ala-Lys(Boc)-OH (585 mg, 1.23 mmol, 1 eq.) instead of Boc-aa.sub.1-OH and HCl.H.sub.2N-Ala-OBn (265 mg, 1.23 mmol, 1 eq.). White solid (364 mg, 48%).

[0440] MS (ESI.sup.+): 613.3 [M+H].sup.+; 635.3 [M+Na].sup.+; 651.3 [M+K].sup.+

[0441] Z-Ala-Lys(HCl)-Ala-OBn:

[0442] Using general procedure C starting from Z-Ala-Lys(Boc)-Ala-OBn (364 mg, mmol, 1 eq.). White solid (326 mg, 100%).

[0443] MS (ESI.sup.+): 513.3 [M-HCl+H].sup.+; 535.3 [M-HCl+Na].sup.+; 551.2 [M-HCl+K].sup.+

[0444] Boc-Tyr(OMe)-Ala-OBn:

[0445] Using general procedure A starting from Boc-Tyr(OMe)-OH (1.35 g, 4.57 mmol, 1 eq.) and HCl.H.sub.2N-Ala-OBn (1.084 mg, 5.03 mmol, 1.1 eq.). White solid (863 mg, 83%).

[0446] MS (ESI.sup.+): 457.2 [M+H].sup.+; 479.2 [M+Na].sup.+; 495.2 [M+K].sup.+

[0447] HCl.H.sub.2N-Tyr(OMe)-Ala-OBn:

[0448] Using general procedure C starting from Boc-Tyr(OMe)-Ala-OBn (1.7 g, mmol, 1 eq.). White solid (1.51 g, 100%).

[0449] MS (ESI.sup.+): 357.2 [M-HCl+H].sup.+; 379.2 [M-HCl+Na].sup.+

[0450] Z-Lys(Boc)-Tyr(OMe)-Ala-OBn:

[0451] Using general procedure D starting from HCl.H.sub.2N-Tyr(OMe)-Ala-OBn (1.53 g, 3.89 mmol, 1 eq.) and Z-Lys(Boc)-OH (1.63 g, 4.28 mmol, 1.1 eq.). Pale yellow solid (1.01 g, 40%).

[0452] MS (ESI.sup.+): 720.4 [M+H].sup.+; 766.5 [M+K].sup.+

[0453] Z-Lys(HCl)-Tyr(OMe)-Ala-OBn:

[0454] Using general procedure C starting from Z-Lys(Boc)-Tyr(OMe)-Ala-OBn (970 mg, 1.35 mmol, 1 eq.). Pale yellow solid (880 mg, 100%).

[0455] MS (ESI.sup.+): 619.3 [M-HCl+H].sup.+; 641.3 [M-HCl+Na].sup.+; 657.3 [M-HCl+K].sup.+

[0456] Synthesis of Intermediate Compounds 2 and 4:

##STR00101##

[0457] With R.sub.1=Bn

[0458] To a cooled (78 C.) solution of compound 1 (1.0 g, 1.33 mmol, 1 eq.), obtained from a process described in WO 2012/085221 A1, in anhydrous toluene (13 mL) was added dropwise a solution of diisobutylaluminium hydride (1.2 M in toluene, 1.4 mL, 1.66 mmol, 1.25 eq.) and the reaction mixture was stirred at the same temperature for 6 h 30 under nitrogen atmosphere. The reaction mixture was quenched with ethanol (3 mL), allowed to warm to 20 C., and stirred at this temperature for 15 minutes. A solution of Rochelle's salt (20%, 8 ml) was added, and the mixture was allowed to warm to room temperature and stirred for 1 h. The reaction mixture was extracted with ethyl acetate (330 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give compound 2 (987 mg, colorless oil) which was used in the next step without any further purification.

[0459] Mass (ESI.sup.+): 777.4 [M+Na].sup.+, 793.4 [M+K].sup.+

[0460] With R.sub.1=Me

[0461] The same procedure as above, using compound 3 (3.56 g, 5.26 mmol) described in Bioorganic & Medicinal Chemistry Letters, 2010, 20, 5251-5254 as starting material, leads to compound 4 (3.11 g, pale yellow oil).

[0462] Mass (ESI.sup.+): 701.3 [M+Na].sup.+, 717.3 [M+K].sup.+

[0463] Synthesis of Compounds 15, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39 and 43

##STR00102## ##STR00103##

[0464] Peptide or amino acid condensation onto hemiketal 2 and subsequent reduction: Hemiketal 2 derivative was dissolved in dichloroethane (0.05M) under nitrogen atmosphere. Magnesium sulfate (3 eq.), the peptide or amino acid derivative (0.95 to 1 eq.) and PS-NEt.sub.2 (3.2 mmol.Math.g.sup.1, 2 eq.) were added sequentially. The reaction mixture was stirred 30 min at room temperature and then refluxed for 20 h. After completion of the reaction (monitored by .sup.19F NMR) the reaction mixture was rapidly filtered over a plug of Celite which was washed with dichloroethane (about 10% of the initial quantity). The obtained solution was cooled to 0 C. and sodium triacetoxyborohydride (2 eq.) and acetic acid (1 eq.) were added sequentially under nitrogen atmosphere. After 30 min stirring at 0 C., the reaction mixture was allowed to warm to room temperature and stirred overnight. A saturated aqueous solution of sodium hydrogenocarbonate was slowly added and the mixture was stirred for 2 h. The layers were separated and the aqueous one extracted with dichloromethane. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to yield the crude glycopeptide derivative which was purified by flash column chromatography.

[0465] Compound 15:

[0466] Using general procedure starting from hemiketal 2 (9.63 g, 12.76 mmol, 1 eq.) and Z-Lys(TFA)-Ala-Ala-OBn (7.59 g, 12.12 mmol, 0.95 eq.). White solid (6.71 g, 46%).

[0467] MS (ESI.sup.+): 1205.6 [M+H].sup.+

[0468] Compound 21:

[0469] Using general procedure starting from hemiketal 2 (1.02 g, 1.35 mmol, 1 eq.) and Z-Lys(HCl)-Ala-OBn (751 mg, 1.35 mmol, 1 eq.). Pale yellow oil (1.14 g, 74%).

[0470] MS (ESI.sup.+): 1134.5 [M+H].sup.+; 1156.5 [M+Na].sup.+

[0471] Compound 23:

[0472] Using general procedure starting from hemiketal 2 (1.02 g, 1.35 mmol, 1 eq.) and Z-Lys(HCl)-Gly-Gly-OBn (786 mg, 1.35 mmol, 1 eq.). White solid (1.03 g, 65%).

[0473] MS (ESI.sup.+): 1177.5 [M+H].sup.+; 1199.5 [M+Na].sup.+

[0474] Compound 25:

[0475] Using general procedure starting from hemiketal 2 (1.01 g, 1.34 mmol, eq.) and Z-Lys(TFA)-Ala-Gly-OBn (924 mg, 1.27 mmol, 0.95 eq.). White solid (719 mg, 47%).

[0476] MS (ESI.sup.+): 1191.5 [M+H].sup.+; 1213.5 [M+Na].sup.+; 1229.5 [M+K].sup.+

[0477] Compound 27:

[0478] Using general procedure starting from hemiketal 2 (1.13 g, 1.49 mmol, 1 eq.) and Z-Lys(TFA)-Gly-Ala-OBn (1.02 g, 1.42 mmol, 0.95 eq.). Pale yellow oil (774 mg, 46%).

[0479] MS (ESI.sup.+): 1191.5 [M+H].sup.+; 1213.5 [M+Na].sup.+; 1229.5 [M+K].sup.+

[0480] Compound 29:

[0481] Using general procedure starting from hemiketal 2 (1.07 g, 1.42 mmol, 1 eq.) and Z-Om(HCl)-Ala-Ala-OBn (802 mg, 1.42 mmol, 1 eq.). White solid (1.23 g, 73%).

[0482] MS (ESI.sup.+): 1191.5 [M+H].sup.+

[0483] Compound 31:

[0484] Using general procedure starting from hemiketal 2 (516 mg, 684 mmol, 1 eq.) and Z-Dab(HCl)-Ala-Ala-OBn (413 mg, 684 mmol, 1 eq.). Colorless oil (588 mg, 73%).

[0485] MS (ESI.sup.+): 1177.5 [M+H].sup.+; 1199.5 [M+Na].sup.+; 1215.5 [M+K].sup.+

[0486] Compound 33:

[0487] Using general procedure starting from hemiketal 2 (497 mg, 0.658 mmol, 1 eq.) and Z-Lys(TFA)-Ala-Pro-OBn (408 mg, 0.625 mmol, 0.95 eq.). Pale yellow oil (466 mg, 61%).

[0488] RMN .sup.19F(CDCl.sub.3, 282.5 MHz) (without .sup.1H coupled): 110.1 (d, 2J.sub.F-F=257 Hz), 111.0 (d, .sup.2J.sub.F-F=257 Hz)

[0489] Compound 35:

[0490] Using general procedure starting from hemiketal 2 (396 mg, 0.524 mmol, 1 eq.) and Z-Lys(TFA)-Pro-Ala-OBn (326 mg, 0.498 mmol, 0.95 eq.). White solid (409 mg, 67%).

[0491] MS (ESI.sup.+): 1231.6 [M+H].sup.+

[0492] Compound 37:

[0493] Using general procedure starting from hemiketal 2 (448 mg, 0.594 mmol, 1 eq.) and Z-Ala-Lys(HCl)-Ala-OBn (331 mg, 0.594 mmol, 1 eq.). Colorless oil (470 mg, 66%).

[0494] MS (ESI.sup.+): 1205.6 [M+H].sup.+; 1227.5 [M+Na].sup.+; 1243.5 [M+K].sup.+

[0495] Compound 39:

[0496] Using general procedure starting from hemiketal 2 (1.01 g, 1.34 mmol, 1 eq.) and Z-Lys(HCl)-Tyr(OMe)-Ala-OBn (830 mg, 1.27 mmol, 0.95 eq.). White solid (1.02 g, 59%).

[0497] MS (ESI.sup.+): 1311.6[M+H].sup.+; 1333.6[M+Na].sup.+; 1349.6 [M+K].sup.+

[0498] Compound 43:

[0499] Using general procedure starting from hemiketal 2 (408 mg, 0.54 mmol, 1 eq.) and Z-Lys(HCl)-OBn (255 mg, 0.54 mmol, 1 eq.). Colorless oil (310 mg, 54%).

[0500] MS (ESI.sup.+): 1063.4 [M+H].sup.+

[0501] Synthesis of Compounds 17, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40 and 44

##STR00104## ##STR00105##

[0502] Glycopeptides deprotection using hydrogenolysis: To a solution of protected glycopeptide in tetrahydrofuran (0.025M) under inert atmosphere was added an aqueous solution of hydrochloric acid (1.0M, 3.6 eq.) followed by Pd/C (10% Pd, 0.1 eq.). The flask was purged with vacuum and filled with H.sub.2 (3 times). The reaction mixture was vigorously stirred under H.sub.2 atmosphere for 18 h. After completion of the reaction, the mixture was filtered (Millipore 0.45 m) and the filter was washed with water. The filtrate was concentrated under vacuum and the obtained residue was dissolved in water and freeze dried to give the desired compound.

[0503] Compound 17:

[0504] Using general procedure starting from compound 15 (6.48 g, 5.38 mmol, 1 eq.). White solid (3.21 g, 99%).

[0505] MS (ESI.sup.+): 531.2 [M-2HCl+H].sup.+; 553.2 [M-2HCl+Na].sup.+; 569.2 [M-2HCl+K].sup.+

[0506] Compound 22:

[0507] Using general procedure starting from compound 21 (702 mg, 0.619 mmol, 1 eq.). White solid (330 mg, 100%).

[0508] MS (ESI.sup.+): 460.2 [M-2HCl+H].sup.+; 482.2 [M-2HCl+Na].sup.+; 498.1 [M-2HCl+K].sup.+

[0509] Compound 24:

[0510] Using general procedure starting from compound 23 (700 mg, 0.595 mmol, 1 eq.). White solid (344 mg, 100%).

[0511] MS (ESI.sup.+): 503.2 [M-2HCl+H].sup.+; 525.2 [M-2HCl+Na].sup.+; 541.2 [M-2HCl+K].sup.+

[0512] Compound 26:

[0513] Using general procedure starting from compound 25 (640 mg, 0.537 mmol, 1 eq.). White solid (307 mg, 97%).

[0514] MS (ESI.sup.+): 517.2 [M-2HCl +H].sup.+; 539.2 [M-2HCl +Na].sup.+; 555.2 [M-2HCl +K].sup.+

[0515] Compound 28:

[0516] Using general procedure starting from compound 27 (604 mg, 0.507 mmol, 1 eq.). White solid (290 mg, 97%).

[0517] MS (ESI.sup.+): 517.2 [M-2HCl +H].sup.+; 539.2 [M-2HCl +Na].sup.+; 555.2 [M-2HCl +K].sup.+

[0518] Compound 30:

[0519] Using general procedure starting from compound 29 (700 mg, 0.588 mmol, 1 eq.). White solid (356 mg, 100%).

[0520] MS (ESI.sup.+): 517.2 [M-2HCl +H].sup.+; 539.2 [M-2HCl +Na].sup.+

[0521] Compound 32:

[0522] Using general procedure starting from compound 31 (643 mg, 0.55 mmol, 1 eq.). Pale orange solid (290 mg, 92%).

[0523] MS (ESI.sup.): 501.2 [M-2HCl H].sup.

[0524] Compound 34:

[0525] Using general procedure starting from compound 33 (436 mg, 0.354 mmol, 1 eq.). White solid (227 mg, 100%).

[0526] MS (ESI.sup.+): 557.2 [M-2HCl +H].sup.+

[0527] Compound 36:

[0528] Using general procedure starting from compound 35 (380 mg, 0.309 mmol, 1 eq.). White solid (196 mg, 100%).

[0529] MS (ESI.sup.+): 557.2 [M-2HCl +H].sup.+

[0530] Compound 38:

[0531] Using general procedure starting from compound 37 (470 mg, 0.39 mmol, 1 eq.). White solid (235 mg, 100%).

[0532] MS (ESI.sup.): 529.3 [M-2HCl H].sup.

[0533] Compound 40:

[0534] Using general procedure starting from compound 39 (50 mg, 0.038 mmol, 1 eq.). White solid (21 mg, 78%).

[0535] MS (ESI.sup.): 635.2 [M-2HCl H].sup.

[0536] Compound 44:

[0537] Using general procedure starting from compound 43 (750 mg, 0.705 mmol, 1 eq.). White solid (305 mg, 94%).

[0538] MS (ESI.sup.): 387.1 [M-2HCl H].sup.

[0539] I-2. Particular Procedures

[0540] Synthesis of Intermediate Compounds 5 and 6:

##STR00106##

[0541] With R.sub.1=Bn

[0542] Compound 2 (606 mg, 0.80 mmol, 1 eq.) and N-carboxybenzoyl-ethyl-lysinate (485 mg, 1.61 mmol, 2 eq.) were refluxed in toluene (8 mL) using a Dean-Stark apparatus under nitrogen atmosphere. After 3 h, the mixture is concentrated in vacuo to obtain compound 5 which was engaged in the next step without further purification. .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 110.2 (d, J=263 Hz, 1F); 114.1 (d, J=263 Hz, 1F).

[0543] With R.sub.1=Me

[0544] The same procedure as above, using compound 4 (1.08 g, 1.59 mmol) as starting material, leads to compound 6 which was engaged in the next step without further purification.

[0545] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 109.7 (d, J=263 Hz, 1F); 114.5 (d, J=263 Hz, 1F).

[0546] Synthesis of Compounds 7 and 8:

##STR00107##

[0547] With R.sub.1=Bn

[0548] Sodium cyanoborohydride (151 mg, 2.41 mmol, 3 eq.) was added to a solution of compound 5 (802 mg, 0.803 mmol, 1 eq.) in tetrahydrofuran (1.6 mL) and methanol (7.4 mL) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 h. A saturated aqueous ammonium chloride solution (8 mL) was added and the mixture was extracted with ethyl acetate (315 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography to yield compound 7 (382 mg, 31%, colorless oil).

[0549] Mass (ESI.sup.+): 1001.6 [M+H].sup.+, 1023.5 [M+Na].sup.+, 1039.5 [M+K].sup.+

[0550] With R.sub.1=Me

[0551] The same procedure as above, using compound 6 (1.47 g, 1.59 mmol) as starting material, leads to compound 8 (796 mg, 54%, pale brown oil).

[0552] Mass (ESI.sup.+): 925.3 [M+H].sup.+

[0553] Alternative Synthesis of Compound 8 (R.sub.1=Me) Using a Substitutive Approach:

[0554] Synthesis of Intermediate Compound 9 (R.sub.1=Me):

##STR00108##

[0555] Sodium borohydride (1 g, 26.4 mmol, 7 eq.) was added portionwise to a cooled (0 C.) solution of compound 3 (2.55 g, 3.77 mmol, 1 eq.) in methanol (30 mL) and THF (2.5 mL) under nitrogen atmosphere. The reaction mixture was allowed to warm to room temperature and stirred for 3 h 30. Saturated ammonium chloride solution (10 mL) and brine (10 mL) were added and the mixture was extracted with ethyl acetate (330 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to yield compound 9 (2.35 g, 98%, pale yellow oil).

[0556] Mass (ESI.sup.+): 657.3 [M+Na].sup.+, 673.3 [M+K].sup.+

[0557] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 113.6 (d, J=262 Hz, 1F); 114.6 (d, J=262 Hz, 1F).

[0558] Synthesis of Intermediate Compound 10 (R.sub.1=Me):

##STR00109##

[0559] Pyridine (650 L, 7.97 mmol, 2.2 eq.) and trifluoromethanesulfonic anhydride (1.3 mL, 7.97 mmol, 2.2 eq.) were added successively to a cooled (78 C.) solution of compound 9 (2.3 g, 3.62 mmol, 1 eq.) in dichloromethane (36 mL) under nitrogen atmosphere. The reaction mixture was allowed to warm to room temperature and stirred for 2 h. Water (20 mL) was added and the mixture was extracted with dichloromethane (330 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to yield compound 10 (2.79 g, 100%, pale orange oil).

[0560] Mass (ESI.sup.+): 789.3 [M+Na].sup.+, 805.2 [M+K].sup.+

[0561] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 113.4 (d, J=259 Hz, 1F); 115.6 (d, J=259 Hz, 1F).

[0562] Synthesis of Compound 8 (R.sub.1=Me):

##STR00110##

[0563] To a solution of compound 10 (100 mg, 0.13 mmol, 1 eq.) in dry DMF (0.65 mL) under nitrogen atmosphere, was added a solution of Z-Lys(NH.sub.2)OEt (80 mg, 0.26 mmol, 2 eq.) in dry DMF (0.65 mL) and potassium carbonate (22 mg, 0.261 mmol, 2 eq.). The reaction mixture was stirred for 2 h 30 at room temperature and 15 h at 50 C. After cooling, saturated ammonium chloride solution (5 mL) and brine (5 mL) were added and the mixture was extracted with ethyl acetate (310 mL). The combined organic layers were washed with brine (220 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to yield compound 8 (15 mg, 13%, colorless oil).

[0564] Mass (ESI.sup.+): 925.3 [M+H].sup.+

[0565] Synthesis of Compounds 11 and 12:

##STR00111##

[0566] With R.sub.1=Bn

[0567] A 2N aqueous solution of lithium hydroxide (75 L, 0.015 mmol, 2 eq.) was added dropwise to a cooled (0 C.) solution of compound 7 (75 mg, 0.075 mmol, 1 eq.) in tetrahydrofuran (750 L). The reaction was then allowed to warm to room temperature and stirred for 18 h. A 1N aqueous solution of HCl was added until pH=1. The reaction mixture was extracted with ethyl acetate (38 mL). The combined organic layers were washed with brine (8 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude compound 11 (76 mg, 93%, pale orange solid) which was used in the next step without further purification.

[0568] Mass (ESI.sup.+): 973.3 [M-HCl+H].sup.+, 995.4 [M-HCl+Na].sup.+

[0569] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 106.8 (d, J=261 Hz, 1F); 109.6 (d, J=261 Hz, 1F).

[0570] With R.sub.1=Me

[0571] The same procedure as above, using compound 8 (604 mg, 0.652 mmol) as starting material, leads to compound 12 (578 mg as a pale orange solid, 95%).

[0572] Mass (ESI.sup.+): 897.5 [M-HCl+H].sup.+, 919.5 [M-HCl+Na].sup.+, 935.4 [M-HCl+K].sup.+

[0573] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 106.1 (d, J=263 Hz, 1F); 108.7 (d, J=263 Hz, 1F).

[0574] Synthesis of Intermediate Compounds 13 and 14:

##STR00112##

[0575] With R.sub.1=Bn

[0576] Compound 2 (45 mg, 0.073 mmol, 1 eq.) and Z-Lys(NH.sub.2)AlaAla-OBn (75 mg, 0.15 mmol, 2 eq.) were refluxed in toluene (5 mL) using a Dean-Stark apparatus under nitrogen atmosphere. After completion of the reaction, the mixture is concentrated in vacuo to obtain compound 13 which was engaged in the next step without further purification.

[0577] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 110.3 (d, J=263 Hz, 1F); 114.0 (d, J=263 Hz, 1F).

[0578] With R.sub.1=Me

[0579] The same procedure as above, using compound 4 (200 mg, 0.32 mmol) as starting material, leads to compound 14 which was engaged in the next step without further purification.

[0580] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 109.5 (d, J=264 Hz, 1F); 114.1 (d, J=264 Hz, 1F).

[0581] Alternative Synthesis of Intermediate Compound 13 (R.sub.1=Bn):

##STR00113##

[0582] With R.sub.1=Bn

[0583] Triethylamine (116 L, 0.83 mmol, 1 eq.) was added dropwise to a stirred solution of compound 2 (628 mg, 0.83 mmol, 1 eq.) in dichloromethane (8.3 mL) under nitrogen atmosphere, Z-Lys(NH.sub.3.sup.+TFA.sup.)AlaAla-OBn (521 mg, 0.83 mmol, 1 eq.) and anhydrous magnesium sulfate (151 mg, 1.25 mmol, 1.5 eq.) were successively added and the mixture was stirred at room temperature for 3 h before being filtered. The filter was washed with a small amount of dichloromethane and the filtrate was washed with water (5 mL), dried with anhydrous magnesium sulfate and evaporated to give pure compound 13 (878 mg, 88%, colorless oil).

[0584] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 110.3 (d, J=263 Hz, 1F); 114.0 (d, J=263 Hz, 1F).

[0585] Alternative Synthesis of Compound 15 and Synthesis of Compound 16:

##STR00114##

[0586] With R.sub.1=Bn

[0587] To a solution of compound 11 (347 mg, 0.34 mmol, 1 eq.) in DMF (3.5 mL), under nitrogen atmosphere, was added a solution of TFA.sup.+H.sub.3N-AlaAla-OBn (163 mg, 0.45 mmol, 1.3 eq.) in DMF (3.5 mL), PyBOP (521 mg, 0.722 mmol, 2.1 eq.), N-methylmorpholine (150 L, 13.8 mmol, 4 eq.). The reaction mixture was stirred for 72 hours. Brine (4 mL) was added and the reaction mixture was extracted with ethyl acetate (38 mL). The combined organic layers were washed with an aqueous 10% citric acid solution (8 mL), 1N aqueous sodium hydroxide (8 mL) and brine (28 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography to yield compound 15 (219 mg, 53%, white solid).

[0588] Mass (ESI.sup.+): 1205.4 [M+H].sup.+

[0589] With R.sub.1=Me

[0590] The same procedure as above, using compound 12 (556 mg, 0.595 mmol) as starting material, leads to compound 16 (544 mg as a pale yellow oil, 81%).

[0591] Mass (ESI.sup.+): 1129.4 [M+H].sup.+, 1151.5 [M+Na].sup.+, 1167.5 [M+K].sup.+

[0592] Alternative Synthesis of Compounds 15 and 16:

##STR00115##

[0593] With R.sub.1=Bn

[0594] Sodium cyanoborohydride (32 mg, 0.51 mmol, 6.9 eq.) was added to a solution of compound 13 (88 mg, 0.073 mmol, 1 eq.) in tetrahydrofuran (0.8 mL) and methanol (1.9 mL) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 20 h. A saturated aqueous ammonium chloride solution (10 mL) was added and the mixture was extracted with ethyl acetate (310 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography to yield compound 15 (57 mg, 68%, pale yellow solid).

[0595] Mass (ESI.sup.+): 1205.6 [M+H].sup.+

[0596] With R.sub.1=Me

[0597] The same procedure as above, using compound 14 (364 mg, 0.32 mmol) as starting material, leads to compound 16 (113 mg, 32%, colorless oil).

[0598] Mass (ESI.sup.+): 1129.5 [M+H].sup.+, 1151.5 [M+Na].sup.+, 1167.5 [M+K].sup.+

[0599] Synthesis of Compound 18:

##STR00116##

[0600] Compound 16 (100 mg, 0.089 mmol, 1 eq.) was dissolved in tetrahydrofuran (798 L) and aqueous hydrochloric acid (2 M, 88 L, 2 eq.). The flask was placed under nitrogen atmosphere and Pd/C 10% (30 mg, 0.32 eq.) was added. The flask was then filled with a hydrogen atmosphere and the reaction mixture was stirred for 24 hours before being Millipore filtered and concentrated in vacuo. The residue was dissolved in water, Millipore filtered (0.2 m) and freeze dried to yield compound 18 (55 mg, 100%) as a pale orange solid.

[0601] Mass (ESI.sup.+): 545.3 [M-2HCl+H].sup.+, 562.4 [M-2HCl+NH.sub.4].sup.+

[0602] .sup.19F NMR (CDCl.sub.3, 282.5 MHz) (without H coupled): 109.1 (d, J=258 Hz, 1F); 110.8 (d, J=258 Hz, 1F).

[0603] Synthesis of Compound 19:

##STR00117##

[0604] Compound 17 (323 mg, 0.54 mmol) was dissolved in water (7 mL) and Amberlite IRA-67 (1.3 g, previously washed with water) was added. The reaction mixture was stirred for 3 hours at room temperature before being Millipore filtered (0.2 inm), diluted with water and freeze dried to yield compound 19 (223 mg, 78%).

[0605] Mass (ESI.sup.+): 531.2 [M+H].sup.+, 553.2 [M+Na].sup.+, 569.2 [M+K].sup.+

[0606] Synthesis of Compound 20:

##STR00118##

[0607] Compound 15 (150 mg, 0.124 mmol, 1 eq.) was dissolved in tetrahydrofuran (1.5 mL), water (1.5 mL) and acetic acid (5.3 mL). The flask was placed under nitrogen atmosphere and Pd/C 10% was added (26 mg, 0.2 eq.). The flask was then filled with a hydrogen atmosphere and the reaction mixture was vigorously stirred for 24 hours before being Millipore filtered and concentrated in vacuo. The residue was dissolved in water, Millipore filtered (0.2 m) and freeze dried to yield compound 20 (66 mg, 82%) as a pale yellow solid.

[0608] Mass (ESI.sup.+): 531.2 [M-2AcOH+H].sup.+, 553.2 [M-2AcOH+Na].sup.+, 569.2 [M-2AcOH+K].sup.+

[0609] Alternative Synthesis of Compound 20:

##STR00119##

[0610] Compound 19 was dissolved in water (10.4 mL). AcOH (6 eq., 1 M, 2.34 mL, 2.34 mmol) was added dropwise and the mixture was manually agitated several times during 1 h 30. The mixture was then freeze dried to give compound 20 (244 mg, 0.375 mmol, 96%) as a white solid.

[0611] Mass (ESI.sup.+): 531.2 [M-2AcOH+H].sup.+, 553.2 [M-2AcOH+Na].sup.+, 569.2 [M-2AcOH+K].sup.+

[0612] Synthesis of Compounds 41 and 42:

##STR00120##

[0613] Hemiketal 45 (lg, 1.58 mmol, 1 eq.) was dissolved in dichloroethane (32 mL) under nitrogen atmosphere. Magnesium sulfate (570 mg, 4.73 mmol, 3 eq.), Z-Lys(HCl)-Ala-Ala-OBn (820 mg, 1.50 mmol, 0.95 eq.) and PS-NEt.sub.2 (3.2 mmol.Math.g.sup.1, 980 mg, 3.15 mmol, 2 eq.) were added sequentially. The reaction mixture was stirred 30 min at room temperature and then refluxed for 20 h. After completion of the reaction (monitored by .sup.19F NMR) the reaction mixture was rapidly filter over a plug of Celite which was washed with dichloroethane. The obtained solution was cooled to 0 C. and sodium triacetoxyborohydride (634 mg, 2.99 mmol, 2 eq.) and acetic acid (86 l, 1.50 mmol, 1 eq.) were added sequentially under nitrogen atmosphere. After 30 min stirring at 0 C., the reaction mixture was allowed to warm to room temperature and stirred overnight. A saturated aqueous solution of sodium hydrogenocarbonate was slowly added and the mixture was stirred for 2 h. The layers were separated and the aqueous one extracted with dichloromethane. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to yield the compound 41 as a white solid (611 mg, 37%).

[0614] MS (ESI.sup.+): 1099.4 [M+H].sup.+; 1121.4 [M+Na].sup.+; 1137.4 [M+K].sup.+

##STR00121##

[0615] To a solution of compound 41 (585 mg, 0.45 mmol, 1 eq.) in THF (17.8 mL) under inert atmosphere was added an aqueous solution of hydrochloric acid (1.61 ml, 1.61 mmol, 3.6 eq.) followed by Pd/C (10% Pd, 4.7 mg, 0.045 mmol, 0.1 eq.). The flask was purged with vacuum and filled with H.sub.2 (3 times). The reaction mixture was vigorously stirred under H.sub.2 atmosphere for 18 h. After completion of the reaction, the mixture was filtered (Millipore 0.45 m) and the filter was washed with water. The filtrate was concentrated under vacuum and the obtained residue was dissolved in water and freeze dried to give compound 42 as a pale orange solid (262 mg, 100%).

[0616] MS (ESI.sup.+): 515.3 [M-2HCl +H].sup.+; 537.2 [M-2HCl +Na].sup.+; 553.2 [M-2HCl +K].sup.+

[0617] Synthesis of Intermediate Compound 45:

##STR00122##

[0618] To a cooled (78 C.) solution of difluoroester (2.19 g, 3.39 mmol, 1 eq.), synthesized according to Synlett 2005,17, 2627-2630 and Org. Lett. 2002, 4, 757-759see also WO 2004/014928, WO 2007/125203 and WO 2007/125194, in anhydrous toluene (34 mL) was added a solution of diisobutylaluminium hydride (1.2M in toluene; 3.7 mL; 4.4 mmol; 1.2 eq.) and the resultant mixture was stirred for 3 h at this temperature. The reaction was then quenched with methanol (6 mL) and the solution was warmed to 20 C. for 15 min. A Rochelle's salt solution (20%, 60 mL) was then added and the solution was vigorously stirred for 30 min. The mixture was then extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over magnesium sulfate, filtered and evaporated in vacuo to give compound 45 as a colorless oil (2.15 g, 93%).

[0619] Mass (ESI.sup.+): 652.2 [M+H.sub.2O].sup.+

[0620] IIBiological Activity:

[0621] II-1. Effect of Glycopeptide 17 on the Preservation/Protection of Neonatal Skin Fibroblasts Under Starvation Conditions. Evaluation by Trypan Blue Exclusion Method.

[0622] Materials and Methods

[0623] Subculturing [0624] The neonatal skin fibroblasts (Cell line: CCD-27SK, ATCC number CRL-1475) were grown with DMEM medium supplemented with Fetal Bovine Serum 10% final, antibiotics (Penicillin/Streptomycin) 1% final and Amphotericin B 0.1% final. [0625] Fibroblasts were grown in 75 cm.sup.2 culture flask to 80% confluence, in 37 C. and 10% CO.sub.2 incubator. The medium was changed every two days by 37 C. preheated fresh medium.

[0626] Starvation Medium [0627] This medium was composed of 45% subculturing medium without Fetal Bovine Serum mixed with 55% of Phosphate Buffer Saline 1 containing EDTA (final concentration of 0.45 mM). This was referred to as serum-free medium.

[0628] Product Preparation [0629] Compound 17 (MM=603.4 g/mol) was diluted in starvation medium to 5 mg/ml final and pH was adjusted at 7.4 by addition of NaOH 1N.

[0630] General Experimental Procedure

[0631] Assays on 96 Well Plates [0632] Fibroblast cells were concentrated to 2.10.sup.5 cells/ml and 100 l of cell suspension was added in wells of a 96-well plate and incubated in 37 C. and 10% CO.sub.2 incubator for 4 hours. [0633] After cell adhesion the medium was changed and plates were incubated (37 C.-10% CO.sub.2) to perform the assay as follow: [0634] One plate for each sampling times: D0, D3, D4, D5, D6, D7, D10, D12 days [0635] Three wells for each condition (triplicate count) added with 120 l of culture medium (surviving control), starvation medium (serum-free control) or compound 17 solution at 5 mg/ml.

[0636] Viability Assay [0637] Cell Viability was evaluated by the Trypan blue exclusion technique based on the principle that live cells possess intact cell membranes that exclude the Trypan blue dye. So, only the dead cells are blue at microscopic observation. [0638] For sampling, 110 l of Trypan Blue (SIGMA T8154) was added to 110 l of trypsinated cell suspension of matching well for counting. [0639] 200 l of the trypan blue/cell mixture are dropped to a hemacytometer. Cells are counting by using a Neubauer-counting chamber. The unstained (viable) and stained (nonviable) cells are counted separately on 9 areas of 1 mm.sup.2 forming a large square of 9 mm.sup.2 and added to obtain the total number of cells per sample. An average of three counts was used to calculate the viability percentage as:

[Number of viable cells/total number of cells]*100 [0640] The cell viability percentages from cultures under starvation conditions were compared with control culture for several days after their addition (D0, D3, D4, D5, D6, D7, D10, D12).

[0641] Results

[0642] The results were plotted in the histogram of FIG. 1 which represents the evolution of fibroblasts viability in vitro during a 12 days period, as well as on the table 1 below.

TABLE-US-00001 TABLE 1 Percentage of viability for 12 days Culture conditions D 0 D 3 D 4 D 5 D 6 D 7 D 10 D 12 Surviving Control 97.43 1.30 95.69 2.97 96.16 1.27 93.49 1.68 91.10 2.24 91.29 0.90 95.21 1.37 90.33 2.46 Serum-Free Control 96.55 1.22 92.73 1.03 76.94 4.03 59.07 5.86 33.54 3.22 15.31 3.46 0.00 0.00 0.00 0.00 Compound 17 96.69 1.85 91.03 1.88 88.98 2.48 86.65 4.26 85.53 1.57 83.20 3.72 80.62 3.72 77.03 2.02 at 5 mg/mL in serum free medium

[0643] The surviving control showed a very good viability between D0 and D12 (viability>90%).

[0644] The cells in starvation medium (serum-free control) showed a clear decrease of their viability from D5 (59%); this phenomenon was greatly accentuated at D6 (34%), D7 (15%) to reach 100% of mortality at D10.

[0645] The viability of cells cultured in starvation medium but treated with compound 17 at 5 mg/ml, decreased very slightly between D0 (97%) and D12 (77%) and remained clearly higher than the serum-free control. Thus compound 17 showed a significant preservative/protective effect on skin fibroblasts since cells have been maintained in a healthy state under unfavorable conditions for growth.

[0646] II-2. Effect of Glycopeptides on the Preservation/Protection of Human Dermis Fibroblasts or Epithelial Nasal Cells in Different Stress Conditions.

[0647] Different stress conditions have been tested to evaluate the preservative/protective effect of several compounds on 2 types of cells: human skin fibroblasts (deprivation, UV stress, oxidative stress or bacterial stress) or human nasal epithelial cells (deprivation). The different protocols are described hereafter.

[0648] Materials and Methods

[0649] Subculturing [0650] The human neonatal skin fibroblasts (cell line: CCD-27SK, ATCC CRL-1475) were grown with DMEM medium supplemented with FBS 10% final, antibiotics (mix Dutscher L0010, Penicillin/Streptomycin/Amphotericin B) 1% final and L-Glutamine 1% final. The human nasal epithelial cells (cell line: RPMI2650, ATCC CCL-30) were grown with MEM medium supplemented with 10% of FBS, 1% of antibiotics and 1% of L-Glutamine. [0651] Cells were grown in 75 cm.sup.2 culture flasks, in 37 C. and 10% CO.sub.2 incubator, until 80% confluence and then subcultured in 3 flasks. The medium was changed every two days by 37 C. preheated fresh medium.

[0652] Assay Media [0653] For starvation assay, the medium was composed of 45% subculturing medium, devoid of FBS, mixed with 55% of PBS 1 containing EDTA (final concentration of 0.45 mM). This was referred to as starvation medium. [0654] For oxidative, bacterial and UV stress the medium was composed of 45% subculturing medium mixed with 55% of PBS 1 containing EDTA (final concentration of 0.45 mM).

[0655] Compound Solutions [0656] The tested compounds were solubilized in PBS (10% of total volume) and then diluted in appropriate assay medium at definite final concentration. When necessary, the pH was adjusted at 7.4 by addition of NaOH 1N.

[0657] General Experimental Procedure

[0658] Assays on 24-Well Plates [0659] Cells were concentrated to 1.10.sup.5 cells/ml and 500 l of cell suspension was added in wells of a 24-well plate and incubated in 37 C. and 10% CO.sub.2 incubator. [0660] After 3 days of incubation the medium was removed in order to perform the assay in a way depending on the stress condition. [0661] Several controls were performed i.e. the positive control (no stress) which was considered as 100% of viability and the stress control (stress, no compound) which was compared with the assay (stress+compound) to show a potential preservative effect of the tested compounds. [0662] In the case of bacterial stress, the lysis of cells was compared between 3 assays only (stress+compound at 3 different concentrations).

[0663] Induced stresses were performed as follow:

[0664] Starvation

[0665] Three wells for each condition (triplicate count) were added with 1 ml of culture medium (positive control) or starvation medium (stress control) or starvation medium added with tested compounds at definite concentrations. Plates were incubated at 37 C. with 10% of CO.sub.2 for different times (1, 4, 6, 7 or 8 days) and then the cell viability was analyzed using the neutral red uptake method.

[0666] Oxidative Stress

[0667] Three wells for each condition (triplicate count) were added with 0.8 ml of culture medium (control) or each tested compound solution. Plates were incubated at 37 C. with 10% of CO.sub.2 for 24 h.

[0668] Then oxidative stress was performed as follow: 0.2 ml of solution at 500 M of H.sub.2O.sub.2 was added in wells except for negative control where 0.2 ml of culture medium were added.

[0669] Plates were incubated at 37 C. with 10% of CO.sub.2 for different time of exposure (1 h and 24 h) and then the cell viability was analyzed using the neutral red uptake method.

[0670] UV Stress

[0671] Three wells for each condition (triplicate count) were added with lml of culture medium (control) or each tested compound solution.

[0672] Plates were incubated at 37 C. with 10% of CO.sub.2 for 24 h.

[0673] Then UV stress was performed as follow: plates were subjected to 11 Joules/cm.sup.2 in a UVA CUBE 400 (HONLE UV technology) and the irradiated plates were incubated at 37 C. with 10% of CO.sub.2 for different times (1 h, 3 h and 24 h) and then the cell viability was analyzed using the neutral red uptake method.

[0674] Bacterial Stress

[0675] Preparation of Supernatant of Staphylococcus aureus Culture.

[0676] Precultured of Staphylococcus aureus was performed in Tryptic Soy Broth during 16 hours at 37 C. under stirring.

[0677] After incubation, bacteria were harvested by centrifugation (4000 rpm, 10 minutes) and suspended to obtain OD.sub.620 nm=3.3 in culture medium of cells without antibiotics. Culture was performed during 24 hours at 37 C. under stirring. Then the supernatant was recovered after centrifugation (4000 rpm, 10 minutes) and sterilized by 0.22 m filtration. Antibiotics (mix Dutscher L0010, Penicillin/Streptomycin/Amphotericin B) were added at 1% to respect proportion of culture medium of cells. The supernatant stored at deep freezer, contains all toxins and metabolic wastes produced during bacterial growth.

[0678] Assay.

[0679] Assay was performed as follow: [0680] Supernatant of Staphylococcus aureus culture was diluted (1/2) in culture medium. [0681] Three wells for each condition (triplicate count) were added with 0.5 ml of tested compound solution. Plates were incubated at 37 C. with 10% of CO.sub.2 for 24 hours. [0682] Then the bacterial stress was performed using the diluted supernatant of Staphylococcus aureus culture to stress the cells. Solutions in each well was removed and replaced by 0.5 ml of diluted supernatants and 0.5 ml of 2 concentrated compound solutions. Plates were incubated at 37 C. with 10% of CO.sub.2 for 48 h. [0683] Then the cell lysis was analyzed using the Lactate Dehydrogenase (LDH) bioassay.

[0684] Viability Assay (Neutral Red Uptake)

[0685] The neutral red uptake assay was used for the determination of cell viability. This assay is based on the ability of viable cells to incorporate and bind the supravital dye neutral red in its lysosomes. So, only the viable cells are dyed. At different times after the stress assay (T0), the plates were incubated with neutral red solution for 3.5 hours. The cells are subsequently washed, the dye is extracted in each well and the absorbance is read using a spectrophotometer.

[0686] For sampling, 1 mL of DMEM (without phenol red indicator) with neutral red (OD=0.110) was added to the cells for 3.5 hours (37 C., 10% CO.sub.2). After incubation, the medium was removed, two washes of PBS were realized and 1 mL of extraction solution (absolute ethanol 49%, ultrapure water 49%, glacial acetic acid 2%) was added. Plates were placed 15 minutes on rotary shaker in the dark before reading OD at 540 nm.

[0687] Calculations

[0688] The OD.sub.540 nm average value of positive control was considered as 100% of viability.

[0689] The percentage of viability for each assay was calculated as follow:

% viability assay=(OD.sub.540 nm of tested solutionblank)*100/(OD.sub.540 nm of positive controlblank).

[0690] The cell viability percentages calculated from stressed cultures added with tested compounds were compared with stress-control culture at different times.

[0691] LDH Bioassay

[0692] Revelation of Effect of Molecules by LDH Bioassay

[0693] The cell lysis was determined using a colorimetric assay that quantitatively measures lactate deshydrogenase (LDH), a stable cytosolic enzyme, which is released upon cell lysis.

[0694] After 48 hours of exposure of bacterial stress, a LDH bioassay was performed. The kit CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega) and its protocol was used to determine the concentration of LDH containing in the different well.

[0695] The OD values at 490 nm represented the level of cell lysis. The average of OD.sub.490 nm values of each tested compound solution were compared together.

[0696] Results

[0697] a) Starvation

[0698] The results obtained with different glycopeptides are gathered in tables 2 to 5 depending on compounds or cells which have been tested. [0699] Human neonatal skin fibroblasts

TABLE-US-00002 TABLE 2 preservative effect of compound 17, compound 19 and compound 20 on human fibroblasts culture for 7 days after serum deprivation Percentage of viability for 7 days Culture conditions D 1 D 4 D 7 Positive control (no stress) 100 2.82 100 6.14 100 5.33 Stress Control (starvation 77 0.21 44 0.77 8 0.56 medium) Stress + compound 17 71 0.53 61 3.35 55 1.60 at 10 mg/mL Stress + compound 17 75 1.40 54 2.77 45 2.16 at 5 mg/mL Stress + compound 20 87 0.81 63 1.21 50 2.78 at 5 mg/mL Stress + compound 19 73 1.40 57 2.05 50 3.40 at 5 mg/mL

[0700] As seen in table 2, the cells in starvation medium (stress control) showed a clear decrease of their viability from D1 (77%) to D7 (8%). Likewise the results show that the viability of stressed cells treated with compound 17, compound 19 or compound 20 at 5 mg/ml, decreased slightly between D1 (73 to 87%) and D7 (45 to 50%) but remained clearly higher than the stress control at D7.

[0701] Thus compounds 17, 19 or 20 showed a significant preservative/protective effect on skin fibroblasts since cells have been maintained in a healthy state under unfavorable conditions for growth.

TABLE-US-00003 TABLE 3 preservative effect of compound 44, compound 24, compound 26, compound 28, compound 30, compound 34 and compound 38 on human skin fibroblast culture for 8 days after serum deprivation Percentage of viability for 8 days Culture conditions D4 D8 Positive control (no stress)* 100 2.70 100 3.72 Stress Control (starvation medium) 83 1.21 34 4.67 Stress + compound 44 at 10 mg/mL 111 2.9 102 1.3 Stress + compound 24 at 10 mg/mL 99 3.35 95 1.60 Stress + compound 26 at 10 mg/mL 89 1.21 64 2.78 Stress + compound 28 at 10 mg/mL 109 2.05 108 3.40 Stress + compound 30 at 10 mg/mL 92 1.2 74 0.5 Stress + compound 34 at 10 mg/mL 103 1 103 2 Stress + compound 38 at 10 mg/mL 101 3 88 3 (*in this assay only the positive control was prepared with culture medium supplemented with 5% of FBS instead of 10%)

[0702] As seen in table 3, the cells in starvation medium (stress control) showed a clear decrease of their viability for 8 days (34%). The viability of skin fibroblasts cultured in starvation medium and added with compounds 44, 24, 26, 28, 30, 34 and 38 at 10 mg/ml, remained clearly higher than that of the stress control at D8 (64 to 108% vs 34%).

[0703] These results showed a significant preservative/protective effect of compounds 44, 24, 26, 28, 30, 34 and 38 on skin fibroblasts maintaining cells in a healthy state under unfavorable conditions for growth.

TABLE-US-00004 TABLE 4 preservative effect of compound 17 and compound 22 on human fibroblast culture for 4 days after serum deprivation Percentage of viability for 4 days Culture conditions D4 Positive control (no stress) 100 2.9 Stress Control (serum free medium) 57 1 Stress + compound 17 at 10 mg/mL 116 6.4 Stress + compound 22 at 10 mg/mL 93 2.2

[0704] As seen in table 4, the cells in starvation medium (stress control) showed a clear decrease of their viability for 4 days (57%) whereas the cell viability of stressed cells treated with compound compound 17 or compound 22 at 10 mg/ml remains very high (116% and 93% respectively). These results showed a significant preservative/protective effect of compounds 17 and 22. [0705] Human nasal epithelial cells

TABLE-US-00005 TABLE 5 preservative effect of compound 19 on human nasal epithelial cell culture for 6 days after serum deprivation Percentage of viability for 6 days Culture conditions D4 D6 Positive control (no stress) 100 33.77 100 17 Stress Control (serum free medium) 27 6.40 7 1.87 Stress + compound 19 at 10 mg/mL 52 4.64 26 4.64

[0706] As seen in the table above, the cells in starvation medium (stress control) showed a clear decrease of their viability at D4 (27%) to D6 (7%). Likewise the results show that the viability of stressed cells added with compound 19 at 10 mg/ml decreased for 6 days but remained higher than the stress control (26% vs 7% at D6). Thus compound 19 showed a preservative/protective effect on human nasal epithelial cells.

[0707] b) Oxidative Stress

[0708] The results obtained with compound 19 at different concentration are gathered in table below.

TABLE-US-00006 TABLE 6 human neonatal skin fibroblasts viability in culture for 24 h after adding H.sub.2O.sub.2 Percentage of viability following oxidative stress Culture conditions 1 h 24 h Positive control (no stress) 100 7.21 100 1.3 Stress Control (100 M H.sub.20.sub.2) 89 4.7 74 6.4 Stress + compound 19 at 5 mg/mL 89 2.7 109 2.2 Stress + compound 19 at 10 mg/mL 99 0.5 120 2.1 Stress + compound 19 at 15 mg/mL 110 2.0 127 5.8

[0709] The results in table 6 show that the viability of stressed cells added with compound 19 at 5 mg/ml, 10 mg/ml or 15 mg/ml increased for 24 h and remained highly higher than that of the stress control (109 to 127% respectively vs 74%, at 24 h after the oxidative stress). Thus, compound 19 has shown a preservative/protective effect against oxidative stress which is dose dependant.

[0710] Moreover in culture added with compound 19, the cell viability increased in time that showed a growth/regenerative effect on stressed cells.

[0711] c) UV Stress

[0712] The results obtained with compound 17 and compound 19 at different concentrations are gathered in table 7 below.

TABLE-US-00007 TABLE 7 human neonatal skin fibroblasts viability in culture for 24 h after UV irradiation Percentage of viability following UV stress (11 J/cm.sup.2) Culture conditions 1 h 3 h 24 h Positive control (no stress) 100 7.21 100 7.21 100 1.29 Stress Control (UV 10 3.1 6 1.61 12 5.3 irradiation) irradiation + compound 17 55 4.0 63 4.2 82 7.4 (5 mg/mL) irradiation + compound 17 53 2.4 46 3.2 94 9.0 (10 mg/mL) irradiation + compound 17 50 0.5 50 1.0 84 9.5 (15 mg/mL) irradiation + compound 19 53 6.6 71 5.9 89 14.0 (5 mg/mL) irradiation + compound 19 70 4.0 80 3.9 116 3.9 (10 mg/mL) irradiation + compound 19 67 1.7 80 1.8 112 2.7 (15 mg/mL)

[0713] As seen in the table above, the irradiated cells in the stress control assay showed a clear decrease of their viability from 1 h after the UV irradiation (viability 10%), whereas the viability of stressed cells added with compounds 17 or 19 at 5 mg/ml increased for 24 h and remained highly higher than that of the stress control (82% and 89% respectively vs 12% at 24 h after the UV stress).

[0714] Thus, compounds 17 and 19 showed a preservative/protective effect against UV stress.

[0715] Moreover in culture added with compound 17 or 19, the cell viability increased in time that showed a growth or regenerative effect on stressed cells.

[0716] d) Bacterial Stress

[0717] The values of OD.sub.490 nm obtained with 3 solutions of compound 17 at 5, 10 or 15 mg/mL were compared in the table 8 below.

TABLE-US-00008 TABLE 8 human neonatal skin fibroblasts lysis at 48 h following bacterial stress Lysis following bacterial stress at 48 h Culture conditions (OD.sub.490 nm) 17 (5 mg/mL) 0.333 0.003 17 (10 mg/mL) 0.293 0.002 17 (15 mg/mL) 0.270 0.005

[0718] The OD.sub.490 nm which represented the cell lysis under bacterial stress at 48 h, decreased when the quantity of compound 17 raised from 5 mg/ml to 15 mg/ml. Compound 17 has thus shown a protective effect on cells against bacterial stress.

[0719] IIIComparative Study

[0720] Comparative Stability Studies Between Compound 17 and Compound X (Referred as Compound 15 in WO 2006/059227)

[0721] The aim of this study is to demonstrate the tremendous improvement of the stability towards basic conditions of compound 17 compared to compound X. For that, the following sets of experiments have been carried out.

[0722] In a first set of experiments (experiments 1 to 7 in table 9), X (10 mg, 1.72 10.sup.2 mmol) was reacted with 2 to 8 equivalents of NaOD in a total volume of 355 L of deuterium oxide ([X]=4.8 10.sup.2 mmol.Math.mL.sup.1) during 2 h 30 before being acidified with 1M DCl in D.sub.2O. To be sure that all compounds in solution are in their acidic form, the equivalents of DCl are adjusted to 3 more equivalents of DCl than the amount required to neutralize the solution (n.sub.DCl=n.sub.NaOD+3). The volume was adjusted to 544 L with deuterium oxide.

[0723] In a second set of experiments (experiments 8 to 14 in table 9), 17 (10 mg, 1.66 10.sup.2 mmol) was reacted with 2 to 8 equivalents of NaOD in a total volume of 343 L of deuterium oxide ([17]=4.8 10.sup.2 mmol.Math.mL.sup.1) during 2 h 30 before being acidified with 1M DCl in D.sub.2O. To be sure that all compounds in solution are in their acidic form, the equivalents of DCl are adjusted to 3 more equivalents of DCl than the amount required to neutralize the solution (n.sub.DCl=n.sub.NaOD+3). The volume was adjusted to 526 L with deuterium oxide.

TABLE-US-00009 TABLE 9 Compound X Experiment no 1 2 3 4 5 6 7 neq.sub.NaOD .sub.1M 2 3 4 5 6 7 8 V.sub.1D2O(L) 320 303 286 269 251 234 217 V.sub.NaOD .sub.1M(L) 34 52 69 86 103 120 138 V.sub.reaction(L) 355 355 355 355 355 355 355 V.sub.DCl .sub.1M(L) 86 103 120 138 155 172 189 V.sub.2D2O(L) 103 86 69 52 35 17 0 V.sub.total 544 544 544 544 544 544 544 Compound 17 Experiment no 8 9 10 11 12 13 14 neq.sub.NaOD .sub.1M 2 3 4 5 6 7 8 V.sub.1D2O(L) 310 294 277 260 244 227 211 V.sub.NaOD .sub.1M(L) 33 50 66 83 100 116 133 V.sub.reaction(L) 343 343 343 343 343 343 343 V.sub.DCl .sub.1M(L) 83 100 116 133 149 166 183 V.sub.2D2O(L) 100 83 66 50 33 17 0 V.sub.total 526 526 526 526 526 526 526

[0724] .sup.1H NMR analyses were performed on each experiment. It is why experiments were realized in deuterium oxide and using 1M NaOD in D.sub.2O as base and 1M DCl in D.sub.2O as acid. These .sup.1H NMR analyses allowed showing the absence of degradation of compound 17 and the degradation of compound X as presented on scheme 1 below:

##STR00123##

[0725] .sup.1H NMR allows indeed the determination of the amount of compound X1 formed, by comparison between the integration corresponding to the methylene group of the lysine side chain connected to the nitrogen atom in X (CH.sub.2NGP) (3.3 ppm) and the integration of the same methylene group in X1 (CH.sub.2Lys hydrolyzed) (3.0 ppm). Results are depicted in table 10 below.

TABLE-US-00010 TABLE 10 integration integration CH.sub.2Lys % Experiment neq.sub.NaOD CH.sub.2NGP hydrolysis hydrolyzed Compound Control 0 2 0.01 0.5 X 1 2 2 0.04 2.0 2 3 2 0.10 4.8 3 4 2 0.30 13.0 4 5 2 0.38 16.0 5 6 2 0.47 19.0 6 7 2 0.52 20.6 7 8 2 0.57 22.2 Compound Control 0 2 0 0 17 8 2 2 0 0 9 3 2 0 0 10 4 2 0 0 11 5 2 0 0 12 6 2 0 0 13 7 2 0 0 14 8 2 0 0

[0726] Regarding these results, it appears that compound X is sensitive to basic medium which is not the case for compound 17. Indeed, the initial .sup.1H NMR spectrum of compound 17 (controlFIG. 2) and the .sup.1H NMR spectra of experiments 8 to 14 (.sup.1H NMR spectrum of experiment 14 is presented on FIG. 3) are very similar while comparison of the initial .sup.1H NMR spectrum of compound X (controlFIG. 4) and the .sup.1H NMR and spectra of experiments 1 to 7 (.sup.1H NMR spectrum of experiment 7 is presented on FIG. 5) show the progressive hydrolysis of compound X by the progressive formation of compound X1 (and thus compound X2), with up to 22% hydrolysis in experiment 7 (8 equivalents of NaOD for 2 h 30).

[0727] To confirm the structure of compounds X1 and X2, resulting from hydrolysis of X, the following experiment was realized, in which X (10 mg, 1.72 10.sup.2 mmol) was reacted with 8 equivalents of NaOD in a total volume of 355 L of deuterium oxide ([X]=4.8 10.sup.2 mmol.Math.mL.sup.1) during 3 days before being acidified with 1M DCl in D.sub.2O. In these conditions, almost the totality of compound X is hydrolyzed, the percentage of hydrolysis of compound X being 85% as determined by .sup.1H NMR as evidence in table 11.

TABLE-US-00011 TABLE 11 integration integration CH.sub.2Lys Experiment neq.sub.NaOD CH.sub.2NGP hydrolyzed % hydrolysis A 0 2 0.01 0.5 B 8 2 11.13 85

[0728] The .sup.1H NMR spectra of the solution of experiments A and B are presented respectively on FIG. 4 and FIG. 6.

[0729] The solution of experiment B was analyzed by mass spectrometry (ESI.sup.+ and ESI.sup.). In ESI.sup.+ two ion adducts with mass of 289.2 and 311.2 were detected, corresponding to [X1-2HCl+H].sup.+ and [X1-2HCl+Na].sup.+ respectively (mass spectrum presented on FIG. 7). In ESI.sup. two ion adducts with mass of 273.1 and 287.2 were detected, corresponding to [X2H].sup. and [X1-2HCl H].sup. respectively (mass spectrum presented on FIG. 8).