GOLD (I)-PHOSPHINE 1,2,3-TRIAZOLE DERIVATIVES WITH ANTIBIOTIC PROPERTIES

Abstract

The present invention relates to gold (I)-phosphine 1,2,3-triazole compounds, and their use in a human or animal medicine. The present invention also relates to using such compounds for the prevention and/or treatment of an infection, i.e. inhibitors of growth of Gram-positive and/or Gram-negative bacteria. On another aspect the invention relates to the synthesis of the gold (I)-phosphine compounds of the invention and to their synthesis intermediates. The present invention finds applications in the medical, veterinary and/or chemical fields.

##STR00001##

Claims

1. A gold (I)-phosphine 1,2,3-triazole compound of formula I or I′: ##STR00195## wherein: each R.sup.1 group is identical or different, and independently represents a linear C.sub.1 to C.sub.14 alkyl group that is branched or unbranched, cyclic or non-cyclic, saturated or unsaturated, optionally comprising heteroatom(s) selected from the group consisting of O, N and S, and optionally covalently linked to form a substituted or non-substituted heterocycle, R.sup.2 represents a substituent of general formula II:
—X—([Y].sub.a—[Z].sub.b).sub.c  Formula II in which a is an integer equal to 0, or 2, b is an integer equal to 0, or 2, c is an integer equal to 0, or 2, X is a monovalent, divalent or trivalent C.sub.1 to C.sub.20 group that is branched or unbranched, saturated or unsaturated, substituted or non-substituted, optionally comprising heteroatom(s) selected from the group consisting of O, N and S, and optionally comprising one or more aromatic or non-aromatic, substituted or non-substituted cycle(s) or heterocycle(s), Y independently represents an amino acid residue, Z independently represents —NR.sup.3R′.sup.3, —SO.sub.2NR.sup.3R′.sup.3—OR.sup.3, a beta-lactam derivative, a (fluoro)quinolone derivative, a cycline derivative, an oxazolidinone derivative, a macrolide derivative, a ketolinde derivative, a puromycin derivative, a amonoside derivative, a lincosamide derivative, a sulfamide derivative, a phenicol derivative, a polymyxin derivative, a rifamycin derivative, a glycopeptide derivative or biotin, and R.sup.3 and R′.sup.3 are identical or different and independently represent H or a C.sub.1 to C.sub.18 linear or branched alkyl group, optionally comprising heteroatom(s) selected from the group consisting of O and N and optionally comprising one or more aromatic or non-aromatic, substituted or non-substituted cycle(s) or heterocycle(s), a pharmaceutically acceptable salt or solvate thereof, and/or isotopes thereof, preferably deuterium.

2. The compound according to claim 1, wherein R.sup.1 is selected from the group consisting of methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, phenyl, benzyl, preferably methyl, ethyl, cyclohexyl, phenyl, and i-propyl.

3. The compound according to claim 1, wherein the linker X is selected from the following structures: ##STR00196## wherein each R.sup.4 and R.sup.5 is identical or different and independently represents —H, a halogen, a C.sub.1 to C.sub.5 linear or branched alkyl group, a C.sub.1 to C.sub.5 linear or branched alkyl group, a C.sub.1 to C.sub.5 linear or branched alkyl group, a C.sub.1 to C.sub.5 linear or branched alkoxy group, —OH or —NR.sup.3R.sup.3, R.sup.3 and R′.sup.3 identical or different, independently being H or a C.sub.1 to C.sub.18 linear or branched alkyl group, optionally comprising heteroatom(s) chosen from the group consisting of O and N, R.sup.3 and R′.sup.3 are optionally covalently linked to form a substituted or non-substituted heterocycle, R.sup.4 and R.sup.5 being optionally covalently linked to form a cycle or heterocycle, n is an integer from 1 to 5, ##STR00197## wherein R.sup.6 is one more substituent(s) on the ring and independently represents one or more of —H, halogen(s), C.sub.1 to C.sub.5 linear or branched alkyl group(s), a C.sub.1 to C.sub.5 linear or branched alkoxy group(s), —OH or —NR.sup.10R′.sup.10, R.sup.10 and R′.sup.10 are optionally covalently linked to form a substituted or non-substituted heterocycle, R.sup.6 groups being optionally covalently linked to form a cycle or heterocycle, ##STR00198## wherein each A.sup.1 independently represents N, CH or CR.sup.11, R.sup.11 is one ore more substituent(s) on the ring and independently represents one or more of —H, halogen(s), a C.sub.1 to C.sub.5 linear or branched alkyl group(s), a C.sub.1 to C.sub.5 linear or branched alkoxy group(s), —OH or —NR.sup.12R′.sup.12, R.sup.12 and R′.sup.12 identical or different, independently being H or a C.sub.1 to C.sub.17 linear or branched alkyl group, optionally comprising heteroatom(s) selected from the group consisting of O and N, R.sup.12 and R′.sup.12 are optionally covalently linked to form a substituted or non-substituted heterocycle, R.sup.11 groups being optionally covalently linked to form a cycle or heterocycle, ##STR00199## wherein each R.sup.7 independently represents —H, a halogen, a C.sub.1 to C.sub.5 linear or branched alkyl group, a C.sub.1 to C.sub.5 linear or branched alkoxy group, —OH or —NR.sup.12R′.sup.12, R.sup.12 and R′.sup.12 identical or different, independently being H or a C.sub.1 to C.sub.17 linear or branched alkyl group, optionally comprising heteroatom(s) selected from the group consisting of O and N, R.sup.12 and R′.sup.12 are optionally covalently linked to form a substituted or non-substituted heterocyclyl, R.sup.7 groups being optionally covalently linked to form a cycle or heterocycle, ##STR00200## wherein A.sup.2 represents CH.sub.2, CHR.sup.8, NR.sup.8, O, S or SO.sub.2, R.sup.9 is one or more substituent(s) on the ring and independently represents one or more of —H, halogen(s), C.sub.1 to C.sub.5 linear or branched alkyl group(s), C.sub.1 to C.sub.5 linear or branched alkoxy group(s), —OH or —NR.sup.13R′.sup.13R.sup.13 and R′.sup.13 identical or different, independently being H or a C.sub.1 to C.sub.15 linear or branched alkyl group, consisting of O and N, R.sup.13 and R′.sup.13 are optionally covalently linked to form a substituted or non-substituted heterocycle, R.sup.9 groups being optionally covalently linked to form a cycle or heterocycle, R.sup.8 represents —H or a C.sub.1 to C.sub.5 linear or branched alkyl group; wherein represents the point of attachment to the 1,2,3-triazole and to the Y or Z group.

4. The compound according to claim 1, wherein the amino acid residue Y is selected from the group consisting of natural and synthetic, alpha, beta or gamma amino acid residues.

5. The compound according to claim 4, where the amino acid is selected from the group consisting of arginine, ω-N.sub.2-arginine histidine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, gluatmine, selenocysteine, glycine, proline, alanine, isoleucine, leucine, phenylalanine, tyrosine, tryptophan, valine, phenylglycine, hydroxyphenylglycine, substituted or not substituted, preferably glycine, phenylglycine, hydroxyphenylglycine, ω—NO.sub.2-arginine, aspartic acid, tryptophan, tyrosine and phenylalanine.

6. The compound according to claim 1, wherein the group Z is a beta-lactam derivative selected from the group consisting of penicillins, cephalosporins, and fluoroquinolone.

7. A process of synthesis of a compound according to claim 1 comprising a step of reacting a compound of formula Ia: ##STR00201## with a compound of formula IB: ##STR00202##

8. The method of claim 7, wherein the compound of formula Ia comprises: ##STR00203## wherein R.sup.2 is defined as above.

9. The method of claim 7, wherein the compound of formula Ib comprises: ##STR00204## wherein R.sup.1 is defined as above.

10. (canceled)

11. A The method of treating a bacterial or fungal bacterial or fungal infection, comprising administering the compound of claim 1.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0153] FIG. 1 is a diagram representing the TMB3 human hepatocyte cytotoxicity between 7 to 25 μM (FIG. 1 A) and the ampicillin human hepatocyte cytotoxicity between 7 to 25 μM (control) (FIG. 1B. In the diagram, the abscissa represents the concentration in Log per μM and the ordinate the viability of the cells in percentage compared to the human hepatocyte culture without ampicillin trihydrate or TMB3.

EXAMPLES

Examples Part A: Synthesis Part I

[0154] All compound and intermediate references below are in the context of part I of the examples.

[0155] All trialkylphosphine-gold(I)-azide intermediates and compounds according to the invention in the present disclosure should be understood as compounds where the gold atom is a gold(I) (oxidation state=1), even if Au is not always represented Au(I) in the following structures.

Preparation of trialkylphosphine-gold(I)-azide intermediates

a. Preparation of triethylphosphine-gold(I)-azide (B)

[0156] ##STR00030##

[0157] Chloro(triethylphosphine) gold (I) A (2.00 g, 5.70 mmol, 1.0 eq.) and thallium(I)acethylacetonate (1.77 g, 5.82 mmol, 1.02 eq.) were suspended in dry toluene (130 mL). Reaction mixture was stirred in the dark at RT for 4 h. Reaction mixture was filtered through Celite® pad and washed with a minimum of toluene. Trimethylsilyl azide (0.92 g, 8.00 mmol, 1.06 mL, 1.4 eq.) was added dropwise to the filtrate, followed by dry methanol (16 mL). Reaction mixture was stirred in the dark at RT for 18 h. Precipitate was removed by filtration through Celite® pad and the filtrate was evaporated under vacuum. Residue was precipitated with toluene/n-pentane (1/50). Supernatant was removed and the white precipitate was washed 3 times with n-pentane and dried under vacuum, affording compound B (1.90 g, 92% yield) as a grey solid.

b. Preparation of triphenylphosphine-gold(I)-azide (D)

[0158] ##STR00031##

[0159] The procedure described above, with chloro(triphenylphosphine) gold (I) C (1.10 g, 2.22 mmol, 1.0 eq.), thallium(I)acethylacetonate (0.69 g, 2.27 mmol, 1.02 eq.) and trimethylsilyl azide (0.35 g, 3.08 mmol, 0.41 mL, 1.4 eq.), afford compound D (0.55 g, 50% yield) as a white/pinky solid.

c. Preparation of dimethylphenylphosphine-gold(I)-azide (F)

[0160] ##STR00032##

[0161] The procedure described above, with chloro(dimethylphenylphosphine) gold (I) E (0.25 g, 0.67 mmol, 1.0 eq.), thallium(I)acethylacetonate (0.21 g, 0.69 mmol, 1.02 eq.) and trimethylsilyl azide (0.11 g, 0.94 mmol, 0.13 mL, 1.4 eq.), afford compound F (0.23 g, 90% yield) as a beige solid.

Preparation of Compounds and Intermediates According to the Invention

1. Preparation of TMB3

[0162] ##STR00033##

a. Preparation of TMB2

[0163] ##STR00034##

[0164] Ampicillin trihydrate 1 (1.00 g, 2.48 mmol, 1.0 eq.) was suspended in a mixture of tetrahydrofurane (10.5 mL) and water (4.5 mL). Triethylamine (0.4 mL) was added to reaction mixture in order to adjust pH (8.0).fwdarw.solubilization. Reaction mixture was cooled down at 0° C., and propargylchloroformate 2 (0.29 g, 2.48 mmol, 0.24 mL, 1.0 eq.) was added dropwise. Another portion of triethylamine (0.5 mL) was added to reaction mixture in order to adjust pH (8.0) and reaction mixture was allowed to warm up to RT for 30 minutes. Tetrahydrofurane was removed under vacuum, water (20 mL) was added to the residue and the resulting aqueous solution was extracted with ethyl acetate (3×30 mL). Organics layers were discarded and aqueous layer was acidified with (1N) hydrochloric acid solution. The white precipitate was collected by filtration, affording compound 3 (0.75 g, 60% yield) as a white/yellowish solid.

b. Preparation of TMB3

[0165] ##STR00035##

[0166] Intermediate TMB2 3 (250 mg, 0.58 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (414 mg, 1.16 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (60 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and 450 mg of the residue were purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.20/80 (20CV)] followed by a direct lyophilization, affording compound 4 (130 mg, 28% yield) as a white solid (TMB3). In some cases, a semi-preparative HPLC was necessary in order to obtain compound 4 with a satisfactory purity.

II. Preparation of TMB1

[0167] ##STR00036##

a. Preparation of Intermediate 6

[0168] ##STR00037##

[0169] D-Phenylglycine 5 (1.00 g, 6.61 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (1.06 g, 26.50 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 2 (1.57 g, 13.22 mmol, 1.29 mL, 2.0 eq.) was added dropwise. Reaction mixture was allowed to warm up to RT for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12CV)] affording compound 6 (1.09 g, 71% yield) as a white solid.

b. Preparation of Compound 7

[0170] ##STR00038##

[0171] Compound 6 (250 mg, 1.07 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (383 mg, 1.07 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the 250 mg of the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV), 100/0.fwdarw.0/100 (20 CV) then 0/100.fwdarw.0/100 (2 CV)] followed by a direct lyophilization, affording compound 7 (170 mg, 27% yield) as a white solid (TMB1).

III. Preparation of TMB5

[0172] ##STR00039##

a. Preparation of Intermediate 12

[0173] ##STR00040##

[0174] Pentynoic acid 8 (1.00 g, 10.19 mmol, 1.0 eq.) was solubilized in dichloromethane (20 mL). 3 drops of dimethylformamide, followed by oxallyl chloride 9 (1.42 g, 11.21 mmol, 0.95 mL, 1.1 eq.) were added dropwise to the reaction mixture. After 1 h of stirring at RT, benzyl alcohol 11 (1.87 g, 17.32 mmol, 1.79 mL, 1.7 eq.) was added and the stirring at RT was continued for 18 h. Reaction was quenched by addition of a saturated solution of NaHCO.sub.3 (20 mL), and the resulting mixture was extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.0/100 (12 CV) then 0/100.fwdarw.0/100 (3 CV)] affording compound 12 in mixture with starting benzyl alcohol 11 (2.41 g, yield not determined) as a colorless oil which became partially solid after overtime. The mixture was used “as is” in the following step.

b. Preparation of Intermediate 14

[0175] ##STR00041##

[0176] Compound 12 (in mixture with benzyl alcohol 11) (1.20 g of the mixture, 5.10 mmol estimated, 1.0 eq.) was solubilized in toluene (20 mL). The solution was degassed by argon bubbling for 30 min. Benzyl azide 13 (1.36 g, 10.20 mmol, 1.06 mL, 2.0 eq.) was added to the reaction mixture and argon bubbling was continued for additional 15 min. Chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II) (97 mg, 0.25 mmol, 0.05 eq.) was added, vessel was sealed and reaction mixture was stirred at 80° C. for 18 h. Reaction was cooled down to RT and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.0/100 (15 CV) then 0/100.fwdarw.0/100 (3 CV)] affording compound 14 (1.06 g, 65% yield) as a brown solid.

c. Preparation of Intermediate 15

[0177] ##STR00042##

[0178] Compound 14 (1.06 g, 3.30 mmol, 1.0 eq.) was solubilized in methanol (100 mL). After Palladium (10% wet on carbon) addition, reaction mixture was hydrogenated (H.sub.2, 5 bars) for 12 h at 10-15° C. Reaction mixture was filtered through Celite® pad and washed with methanol. Solvents were evaporated under vacuum, affording compound 15 (460 mg, 99% yield) as a white/pinky solid.

d. Preparation of Intermediate 17

[0179] ##STR00043##

[0180] Compound 15 (380 mg, 2.62 mmol, 1.0 eq.) was solubilized in dichloromethane (10 mL). Pentafluorophenol 16 (471 mg, 2.56 mmol, 0.95 eq.) and EDCl (491 mg, 2.56 mmol, 0.95 eq.) were added to the reaction mixture and stirred at RT for 2 h. Solvents were evaporated under vacuum, affording crude compound 17 as a sticky brownish solid which can be used “as is” in the following step. Formation of compound 17 is confirmed by LCMS.

e. Preparation of Compound 18

[0181] ##STR00044##

[0182] Crude pentafluorophenyl ester 17 (2.56 mmol estimated, 1.1 eq.) was suspended in tetrahydrofurane (20 mL). Ampicillin trihydrate 1 (939 mg, 2.33 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (707 mg, 6.99 mmol, 0.97 mL, 3.0 eq.). Ampicillin solution was added to the pentafluorophenyl ester 17 suspension and was stirred 1 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting white precipitate was collected by filtration over fritté. Water was removed by trituration in toluene and solvents evaporation under vacuum (×3). The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.0/100 (30 CV) then 0/100.fwdarw.0/100 (3 CV)] followed by a direct lyophilization, affording compound 18 (90 mg, 8% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done, followed by a direct lyophilization, affording compound 18 (21 mg, 2% yield) as a white solid (TMB5).

IV. Preparation of TMB6

[0183] ##STR00045##

a. Preparation of Intermediate 19

[0184] ##STR00046##

[0185] Pentynoic acid 8 (1.50 g, 15.29 mmol, 1.0 eq.) was solubilized in dichloromethane (30 mL). 3 drops of dimethylformamide, followed by oxallyl chloride 9 (2.13 g, 16.82 mmol, 1.1 mL, 1.42 mL, 1.1 eq.) were added dropwise to the reaction mixture. After 2 h of stirring at RT, absolute ethanol (30 mL) was added and the stirring at RT was continued for 18 h. Reaction was quenched by addition of a saturated solution of NaHCO.sub.3 (20 mL). The resulting mixture was extracted with dichloromethane (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum, affording compound 19 (1.10 g, 58% yield) as a colorless oil.

b. Preparation of Intermediate 20

[0186] ##STR00047##

[0187] Pentynoic acid ethyl ester 19 (1.10 g, 8.72 mmol, 1.0 eq.) was solubilized in toluene (20 mL). The solution was degassed by argon bubbling for 30 min. Benzyl azide 13 (1.51 g, 11.34 mmol, 1.42 mL, 1.3 eq.) was added to the reaction mixture and argon bubbling was continued for additional 15 minutes. Chloro(pentamethylcyclopentadienyl) (cyclooctadiene) ruthenium(II) (166 mg, 0.44 mmol, 0.05 eq.) was added, vessel was sealed and reaction mixture was stirred at 80° C. for 18 h. Reaction was cooled down to RT and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV) then 100/0.fwdarw.0/100 (12 CV)] affording compound 20 (1.94 g, 86% yield) as a brown oil.

c. Preparation of Intermediate 21

[0188] ##STR00048##

[0189] Compound 20 (1.94 g, 7.48 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (100 mL) and water (20 mL). Lithium hydroxide hydrate (470 mg, 11.22 mmol, 1.5 eq.) was added to the reaction mixture which was stirred at RT for 72 h. Solvents were evaporated under vacuum and the resulting aqueous solution was was acidified with (1N) hydrochloric acid solution. After extraction with ethyl acetate (3×100 mL), drying over magnesium sulfate and solvents evaporation under vacuum, the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV) then 0/100.fwdarw.0/100 (5 CV)] affording compound 21 (720 mg, 40% yield) as a white/yellowish powder.

d. Preparation of Intermediate 22

[0190] ##STR00049##

[0191] Compound 21 (440 mg, 1.90 mmol, 1.0 eq.) was solubilized in dichloromethane (10 mL). Pentafluorophenol 16 (350 mg, 1.90 mmol, 1.0 eq.) and EDCl (364 mg, 1.90 mmol, 1.0 eq.) were added to the reaction mixture and were stirred at RT for 2 h. Solvents were evaporated under vacuum, affording crude compound 22 as a sticky brownish solid which can be used “as is” in the following step. Formation of compound 22 is confirmed by LCMS.

e. Preparation of Compound 23

[0192] ##STR00050##

[0193] Crude pentafluorophenyl ester 22 (1.90 mmol estimated, 1.1 eq.) was suspended in tetrahydrofurane (20 mL). Ampicillin trihydrate 1 (697 mg, 1.73 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (525 mg, 5.19 mmol, 0.72 mL, 3.0 eq.). Ampicillin solution was added to the pentafluorophenyl ester 22 suspension and was stirred 1 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting gum was isolated by supernatant removal. Water was removed by trituration in ethyl acetate/tetrahydrofurane/toluene mixture and solvents evaporation under vacuum (×2). The residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV) then 100/0.fwdarw.50/50 (30 CV)] followed by a direct lyophilization, affording compound 23 (230 mg, 24% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done on 100 mg of the residue, followed by a direct lyophilization, affording compound 23 (55 mg, 19% yield) as a white solid (TMB6).

V. Preparation of TMB7

[0194] ##STR00051##

a. Preparation of Intermediate 25

[0195] ##STR00052##

[0196] Ethyl propiolate 24 (1.10 g, 10.19 mmol, 1.0 eq.) was solubilized in toluene (20 mL). The solution was degassed by argon bubbling for 30 min. Benzyl azide 13 (1.76 g, 13.25 mmol, 1.66 mL, 1.3 eq.) was added to the reaction mixture and argon bubbling was continued for additional 15 min. Chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II) (193 mg, 0.51 mmol, 0.05 eq.) was added, vessel was sealed and reaction mixture was stirred at 85° C. for 24 h. Reaction was cooled down to RT and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 25 (480 mg, 20% yield) as a dark red oil.

b. Preparation of Intermediate 26

[0197] ##STR00053##

[0198] A solution of 1.0M lithium aluminium hydride in tetrahydrofurane (8.30 mmol, 8.30 mL, 4.0 eq.) was diluted in dry tetrahydrofurane (50 mL) and cooled down to −10° C., under argon atmosphere. Compound 25 (480 mg, 2.07 mmol, 1.0 eq.) was solubilized in dry tetrahydrofurane (50 mL). This solution was added dropwise to the cooled solution of lithium aluminium hydride. Reaction mixture was allowed to warm up to RT and was stirred for 2 h. After cooling down at 0° C. a saturated solution of Na.sub.2SO.sub.4 was carefully added to the mixture. The resulting white precipitate/gum was removed by filtration over cotton pad, and the filtrate was extracted ethyl acetate (3×300 mL), dried over magnesium sulfate and solvents were evaporated under vacuum, affording compound 26 (400 mg, 99% yield) as a brown oil.

c. Preparation of Intermediate 28

[0199] ##STR00054##

[0200] p-Nitrophenyl-chloroformate 27 (426 mg, 2.11 mmol, 1.0 eq.) was solubilized in tetrahydrofurane (30 mL). The solution was cooled down at 0° C. under argon atmosphere. Triethylamine (427 mg, 4.22 mmol, 0.59 mL, 2.0 eq.) was added to the reaction mixture followed by compound 26 (400 mg, 2.11 mmol, 1.0 eq.) in solution in tetrahydrofurane (10 mL). Reaction mixture was stirred for 3 h at RT and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (2 CV), 100/0.fwdarw.50/50 (15 CV), then 50/50.fwdarw.50/50 (3 CV)] affording compound 28 (180 mg, 24% yield) as a colorless oil.

d. Preparation of Compound 29

[0201] ##STR00055##

[0202] Compound 28 (180 mg, 0.51 mmol, 1.0 eq.) was suspended in tetrahydrofurane (20 mL). Ampicillin trihydrate 1 (205 mg, 0.51 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (155 mg, 1.53 mmol, 0.21 mL, 3.0 eq.). Ampicillin solution was added to the compound 28 solution and was stirred 1 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting white precipitate was isolated by filtration over fritté. Water was removed by trituration in tetrahydrofurane/toluene mixture and solvents evaporation under vacuum (×2). The gummy residue was purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) followed by a direct lyophilization, affording compound 29 (43 mg, 15% yield) as a white solid (TMB7).

VI. Preparation of TMB8

[0203] ##STR00056##

a. Preparation of Intermediate 31

[0204] ##STR00057##

[0205] (R)-(−)-2-Phenylglycine methyl ester hydrochloride 30 (2.00 g, 9.92 mmol, 1.0 eq.) was stirred at RT for 18 h in methylamine solution 33% in ethanol (30 mL). Solvents were evaporated under vacuum. To the resulting yellow/orange gum was added diethyl ether (100 mL) which was evaporated under vacuum (×3). After trituration in dichloromethane/diethylether (1/10), the resulting precipitate was filtered over fritté to afford compound 31 (1.54 g, 77% yield) as a yellow/brownish powder.

b. Preparation of Intermediate 32

[0206] ##STR00058##

[0207] Intermediate 31 (500 mg, 2.49 mmol, 1.0 eq.) was suspended in dry tetrahydrofurane (20 mL). Dropwise addition of triethylamine (756 mg, 7.47 mmol, 1.04 mL, 3.0 eq.) followed by propargylchloroformate 2 (325 mg, 2.74 mmol, 0.27 mL, 1.1 eq.) and reaction mixture was stirred at RT for 1 h. Solvents were evaporated under vacuum, affording a brown residue which was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: DCM/MeOH; gradient: 100/0.fwdarw.100/0 (2 CV), then 100/0.fwdarw.80/00 (30 CV)] affording compound 32 (100 mg, 16% yield) as a brown powder.

c. Preparation of Compound 33

[0208] ##STR00059##

[0209] Compound 32 (100 mg, 0.41 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (145 mg, 0.41 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (25 mL) and stirred in the dark at RT for 72 h. Solvents were evaporated under vacuum, the residue was triturated in a tetrahydrofurane/n-pentane (1/10) mixture and the white solid was collected by filtration affording compound 33 (210 mg, 85% yield). A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done on the residue, followed by a direct lyophilization, affording compound 33 (60 mg, 24% yield) as a white solid (TMB8).

VII. Preparation of TMB9

[0210] ##STR00060##

a. Preparation of Intermediate 34

[0211] ##STR00061##

[0212] Benzyl azide 13 (1.00 g, 7.51 mmol, 0.94 mL, 1.0 eq.) and pentynoic acid 8 (810 mg, 8.26 mmol, 1.1 eq.) were solubilized in tertio-butyl alcohol (6.5 mL). A 0.4M aqueous solution of copper (II) acetate was prepared by solubilizing copper (II) acetate (72 mg, 0.40 mmol, 0.05 eq.) in water (1 mL). This solution was added to the reaction mixture and stirred at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: DCM/MeOH; gradient: 100/0.fwdarw.90/10 (12 CV)] affording compound 34 (1.10 g, 65% yield) as a white solid.

b. Preparation of Intermediate 35

[0213] ##STR00062##

[0214] Compound 34 (1.10 g, 4.76 mmol, 1.0 eq.) was solubilized in dichloromethane (10 mL). Pentafluorophenol 16 (870 mg, 4.76 mmol, 1.0 eq.) and EDCl (910 mg, 4.76 mmol, 1.0 eq.) were added to the reaction mixture and stirred at RT for 2 h. Solvents were evaporated under vacuum, affording crude compound 35 as a sticky brownish solid which can be used “as is” in the following step. Formation of compound 35 is confirmed by LCMS.

c. Preparation of Compound 36

[0215] ##STR00063##

[0216] Crude pentafluorophenyl ester 35 (4.76 mmol estimated, 1.1 eq.) was suspended in tetrahydrofurane (20 mL). Ampicillin trihydrate 1 (1.74 g, 4.32 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (1.31 g, 12.96 mmol, 1.81 mL, 3.0 eq.). Ampicillin solution was added to the pentafluorophenyl ester 35 suspension and was stirred for 1 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting residue was isolated by supernatant removal. 1.6 g of the residue were purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV) then 100/0.fwdarw.75/25 (30 CV)] followed by a direct lyophilization, affording compound 36 (410 mg, 33% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done on 165 mg of the residue, followed by a direct lyophilization, affording compound 36 (48 mg, 9% yield) as a white solid (TMB9).

VIII. Preparation of TMB10

[0217] ##STR00064##

a. Preparation of Compound 37

[0218] ##STR00065##

[0219] Intermediate TMB2 3 (167 mg, 0.39 mmol, 1.0 eq.) and triphenylphosphine-gold-azide 0 (389 mg, 0.78 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (40 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was splited in half for purification by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.50/50 (30CV)] (×2) followed by a direct lyophilization, affording compound 37 (220 mg, 60% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done on the residue, followed by a direct lyophilization, affording compound 37 (58 mg, 34% yield) as a white solid (TMB10).

IX. Preparation of TMB4

[0220] ##STR00066##

a. Preparation of Intermediate 38

[0221] ##STR00067##

[0222] Pentynoic acid 8 (500 mg, 5.10 mmol, 1.0 eq.) was solubilized in dichloromethane (10 mL). Pentafluorophenol 16 (891 mg, 4.84 mmol, 0.95 eq.) and EDCl (928 mg, 4.84 mmol, 0.95 eq.) were added to the reaction mixture and stirred at RT for 2 h. Solvents were evaporated under vacuum, affording crude compound 38 as a sticky brownish solid which can be used “as is” in the following step. Formation of compound 38 is confirmed by LCMS.

b. Preparation of Intermediate 39

[0223] ##STR00068##

[0224] Crude pentafluorophenyl ester 38 (4.84 mmol estimated, 1.2 eq.) was suspended in tetrahydrofurane (20 mL). Ampicillin trihydrate 1 (1.51 g, 3.74 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (1.13 g, 11.22 mmol, 1.56 mL, 3.0 eq.). Ampicillin solution was added to the pentafluorophenyl ester 38 suspension and was stirred for 1 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting precipitate was isolated by filtration over fritté, washed with water and was directly lyophilized. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV) then 100/0.fwdarw.75/25 (30 CV)] followed by a direct lyophilization, affording compound 39 (380 mg, 24% yield) as a white solid.

c. Preparation of Compound 40

[0225] ##STR00069##

[0226] Intermediate 39 (200 mg, 0.47 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (330 mg, 0.93 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (50 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue split in half for purification by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV) then 100/0.fwdarw.50/50 (30 CV)] (×2) followed by a direct lyophilization, affording compound 40 (140 mg, 38% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done on the residue, followed by a direct lyophilization, affording compound 40 (11 mg, 3% yield) as a white solid (TMB4).

X. Preparation of TMB11

[0227] ##STR00070##

a. Preparation of Compound 41

[0228] ##STR00071##

[0229] Intermediate TMB2 3 (131 mg, 0.30 mmol, 1.0 eq.) and dimethylphenylphosphine-gold-azide F (229 mg, 0.61 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 20 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.20/80 (15CV)] followed by a direct lyophilization, affording compound 41 (94 mg, 39% yield) as a white solid (TMB11).

XI. Preparation of TMB12

[0230] ##STR00072##

a. Preparation of Intermediate 43

[0231] ##STR00073##

[0232] 3-Ethynylbenzoic acid 42 (670 mg, 4.58 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (1.64 g, 4.58 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (50 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was split in half for purification by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV) then 100/0.fwdarw.50/50 (25 CV)] (×2) followed by a direct lyophilization, affording compound 43 (985 mg, 43% yield) as a white solid.

b. Preparation of Intermediate 44

[0233] ##STR00074##

[0234] Intermediate 43 (460 mg, 0.91 mmol, 1.0 eq.) was solubilized in dichloromethane (10 mL). Pentafluorophenol 16 (160 mg, 0.87 mmol, 0.95 eq.) and EDCl (166 mg, 0.87 mmol, 0.95 eq.) were added to the reaction mixture and stirred at RT for 2 h. Solvents were evaporated under vacuum, affording crude compound 44 as a sticky yellow oil which can be used “as is” in the following step. Formation of compound 44 is confirmed by LCMS.

c. Preparation of Compound 45

[0235] ##STR00075##

[0236] Crude pentafluorophenyl ester 44 (0.91 mmol estimated, 1.0 eq.) was suspended in tetrahydrofurane (100 mL). Ampicillin trihydrate 1 (512 mg, 1.27 mmol, 1.4 eq.) was solubilized in a mixture of tetrahydrofurane (10 mL), water (5 mL) and triethylamine (276 mg, 2.73 mmol, 0.38 mL, 3.0 eq.). Ampicillin solution was added to the pentafluorophenyl ester 44 suspension and was stirred 2 h at RT. Solvents were evaporated under vacuum, and the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting precipitate was isolated by supernatant removal, washed with water and directly lyophilized. 200 mg of the residue were purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18-82/18, 10 minutes and 82/18-0/100, 40 minutes) followed by a direct lyophilization, affording compound 45 (14 mg, 6% yield) as a white solid (TMB12).

XII. Preparation of TMB13

[0237] ##STR00076##

[0238] See procedure for preparation of the intermediate 43

XIII. Preparation of TMB14

[0239] ##STR00077##

a. Preparation of Compound 47

[0240] ##STR00078##

[0241] 4-Ethynylbenzoic acid 46 (350 mg, 2.40 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (855 mg, 2.40 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (70 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum, the residue was triturated in a DCM/THF/n-pentane (1/1/10) mixture and the supernatant was removed. The resulting residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV) then 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 47 (170 mg, 11% yield) as a white solid (TMB14).

XIV. Preparation of TMB15

[0242] ##STR00079##

a. Preparation of Intermediate 49

[0243] ##STR00080##

[0244] tertio-Butyl ester D-phenylglycine hydrochloride 48 (500 mg, 2.05 mmol, 1.0 eq.) was suspended in tetrahydrofurane (30 mL). Solubilization occurs after triethylamine addition (830 mg, 8.20 mmol, 1.14 mL, 4.0 eq.). Propargylchloroformate 2 (486 mg, 4.10 mmol, 0.40 mL, 2.0 eq.) was added dropwise and reaction mixture was stirred at RT for 3 h. Reaction mixture was filtered through cotton pad and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (5CV), 100/0.fwdarw.50/50 (15CV), then 50/50.fwdarw.50/50 (2CV)] affording compound 49 (460 mg, 78% yield) as a colorless oil.

b. Preparation of Compound 50

[0245] ##STR00081##

[0246] Compound 49 (250 mg, 0.86 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (307 mg, 0.86 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.40/60 (25CV)] affording compound 50 (160 mg, 29% yield) as a white powder (TMB15).

XV. Preparation of TMB16

[0247] ##STR00082##

a. Preparation of Compound 51

[0248] ##STR00083##

[0249] Compound 6 (250 mg, 1.07 mmol, 1.0 eq.) and triphenylphosphine-gold-azide D (536 mg, 1.07 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5 CV), then 100/0.fwdarw.40/60 (25 CV)] followed by a direct lyophilization, affording compound 51 (437 mg, 56% yield) as a white solid (TMB16).

XVI. Preparation of TMB17

[0250] ##STR00084##

a. Preparation of Intermediate 53

[0251] ##STR00085##

[0252] Glycine 52 (1.0 g, 13.32 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (2.13 g, 53.28 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., propargylchloroformate 2 (3.16 g, 26.64 mmol, 2.60 mL, 2.0 eq.) was added dropwise and reaction mixture was allowed to warm up to RT for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum, affording crude intermediate 53 (2.14 g, 99% yield) as a colorless/pinky solid which can be used in the next step without purification.

b. Preparation of Intermediate 54

[0253] ##STR00086##

[0254] Compound 53 (200 mg, 1.27 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (454 mg, 1.27 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), 100/0.fwdarw.80/20 (20CV), then 80/20.fwdarw.80/20 (5CV)], followed by a direct lyophilization, affording compound 54 (110 mg, 17% yield) as a white powder. A second purification by semi-preparative HPLC was done (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) followed by a direct lyophilization, affording compound 54 (30 mg, 5% yield) as a white powder (TMB18).

XVII. Preparation of TMB19

[0255] ##STR00087##

a. Preparation of Compound 55

[0256] ##STR00088##

[0257] Pentynoic acid 8 (150 mg, 1.53 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (546 mg, 1.53 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.50/50 (12CV)] followed by a direct lyophilization, affording compound 55 (270 mg, 39% yield) as a white powder (TMB19).

XVIII. Preparation of TMB20

[0258] ##STR00089##

a. Preparation of Compound 56

[0259] ##STR00090##

[0260] Crude pentafluorophenyl ester 44 (0.94 mmol estimated, 1.0 eq.) was suspended in tetrahydrofurane (20 mL). D-Phenylglycine 5 (142 mg, 0.94 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (285 mg, 2.82 mmol, 0.39 mL, 3.0 eq.). D-Phenyglycine solution was added to the pentafluorophenyl ester 44 suspension and was stirred 2 h at RT. Solvents were evaporated under vacuum, and the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.40/60 (25CV)] followed by a direct lyophilization, affording compound 56 (60 mg, 10% yield) as a white powder. A second purification by semi-preparative HPLC was done (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) followed by a direct lyophilization, affording compound 56 (10 mg, 2% yield) as a white powder (TMB20).

XIX. Preparation of TMB21

[0261] ##STR00091##

a. Preparation of Intermediate 59

[0262] ##STR00092##

[0263] Pentynoic acid 8 (1.00 g, 10.19 mmol, 1.0 eq.) was solubilized in tetrahydrofurane (20 mL) and cooled down at −10° C. under argon atmosphere. Triethylamine (3.09 g, 30.57 mmol, 4.26 mL, 3.0 eq.) was added dropwise, followed by methylchloroformate 57 (1.06 g, 11.21 mmol, 0.87 mL, 1.1 eq.). Reaction mixture was stirred at −10° C. for 1 h. (R)-(−)-2-Phenylglycine methyl ester hydrochloride 30 (2.05 g, 10.19 mmol, 1.0 eq.) was added portionwise to the reaction mixture and the stirring was continued for 18 h at RT. Solvents were evaporated under vacuum, water (50 mL) was added to the residue and the resulting aqueous mixture was extracted with ethyl acetate (3×100 mL). Organic phase was dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3CV), 100/0-0/100 (15CV), then 0/100.fwdarw.0/100 (3CV)] affording compound 59 (1.83 g, 73% yield) as a colorless oil.

b. Preparation of Intermediate 60

[0264] ##STR00093##

[0265] Compound 59 (1.83 g, 7.46 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL) and water (4 mL). Lithium hydroxide hydrate (626 mg, 14.92 mmol, 2.0 eq.) was added to the reaction mixture was stirred at RT for 1 h. Solvents were evaporated under vacuum and the resulting aqueous solution was was acidified with (1N) hydrochloric acid solution. After extraction with ethyl acetate (3×100 mL), drying over magnesium sulfate and solvents evaporation under vacuum, the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3CV), 100/0.fwdarw.0/100 (12 CV) then 0/100.fwdarw.0/100 (3 CV)] affording compound 60 (1.25 g, 72% yield) as a white powder.

c. Preparation of Compound 61

[0266] ##STR00094##

[0267] Compound 60 (250 mg, 1.08 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (771 mg, 2.16 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at RT for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (4 CV), 100/0.fwdarw.80/20 (15CV) then 80/20.fwdarw.80/20 (2 CV)] followed by a direct lyophilization, affording compound 61 (200 mg, 31% yield) as a white solid (TMB21).

XX. Preparation of TMB22

[0268] ##STR00095##

a. Preparation of Intermediate 62

[0269] ##STR00096##

[0270] Intermediate 47 (300 mg, 0.60 mmol, 1.0 eq.) was solubilized in dichloromethane (30 mL). Pentafluorophenol 16 (104 mg, 0.57 mmol, 0.95 eq.) and EDCl (109 mg, 0.57 mmol, 0.95 eq.) were added to the reaction mixture and stirred at RT for 18 h. Solvents were evaporated under vacuum, affording crude compound 62 as a sticky yellow oil which can be used “as is” in the following step. Formation of compound 62 is confirmed by LCMS.

b. Preparation of Compound 63

[0271] ##STR00097##

[0272] Crude pentafluorophenyl ester 62 (0.60 mmol estimated, 1.0 eq.) was suspended in tetrahydrofurane (12 mL). Ampicillin trihydrate 1 (475 mg, 1.18 mmol, 2.0 eq.) was solubilized in a mixture of tetrahydrofurane (12 mL), water (6 mL) and triethylamine (238 mg, 2.35 mmol, 0.32 mL, 3.9 eq.). Ampicillin solution was added to the pentafluorophenyl ester 62 suspension and was stirred 3 h at RT. Solvents were evaporated under vacuum, the residual aqueous phase cooled down to 0° C. and was acidified with (1N) hydrochloric acid solution. The resulting yellow gum was isolated by supernatant removal and directly lyophilized. The residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.35/65 (12 CV)] affording compound 63 (90 mg, 10% yield) as a white powder. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done, followed by a direct lyophilization, affording compound 63 (6 mg, 1% yield) as a white solid (TMB22).

XXI. Preparation of TMB23

[0273] ##STR00098##

a. Preparation of Intermediate 64

[0274] ##STR00099##

[0275] Crude pentafluorophenyl ester 62 (0.34 mmol estimated, 1.0 eq.) was suspended in tetrahydrofurane (20 mL). (R)-(−)-2-Phenylglycine methyl ester hydrochloride 30 (83 mg, 0.41 mmol, 1.2 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL) and triethylamine (285 mg, 1.02 mmol, 0.39 mL, 3.0 eq.). (R)-(−)-2-Phenylglycine methyl ester hydrochloride 30 solution was added to the pentafluorophenyl ester 62 suspension and was stirred 18 h at RT. Reaction mixture was filtered through cotton pad and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.75/25 (7CV)] followed by a direct lyophilization, affording compound 64 (150 mg, 68% yield) as a white powder.

b. Preparation of Compound 65

[0276] ##STR00100##

[0277] Compound 64 (150 mg, 0.23 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL) and water (2 mL). Lithium hydroxide hydrate (29 mg, 0.69 mmol, 3.0 eq.) was added to the reaction mixture was stirred at RT for 18 h. Solvents were evaporated under vacuum and the resulting aqueous solution was was acidified with (1N) hydrochloric acid solution. The resulting acidic aqueous phase was directly purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), 100/0.fwdarw.60/40 (9CV) then 60/40.fwdarw.60/40 (2CV)] followed by a direct lyophilization, affording compound 65 (40 mg, 27% yield) as a white powder. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done, followed by a direct lyophilization, affording compound 65 (11 mg, 7% yield) as a white solid (TMB23).

XXII. Preparation of TMB24

[0278] ##STR00101##

a. Preparation Intermediate 67

[0279] ##STR00102##

[0280] Propargylamine 66 (1.00 g, 18.2 mmol, 1.16 mL, 1.0 eq.) was solubilized in tetrahydrofurane (30 mL) and cooled down at 0° C. under argon atmosphere. A solution of p-nitrophenyl-chloroformate 27 (3.7 g, 18.2 mmol, 1.0 eq.) in tetrahydrofurane (10 mL) was added dropwise to reaction mixture and was allowed to warm-up to RT. Stirring was continued for 3 h. Solvents were removed under vacuum, and the residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.55/45 (12CV), then 55/45.fwdarw.20/80 (12CV)] affording compound 67 in mixture with residual p-nitrophenol (1/1) (3.3 g) as a yellow powder.

b. Preparation Intermediate 68

[0281] ##STR00103##

[0282] Ampicillin trihydrate 1 (1.29 g, 3.20 mmol, 1.0 eq.) was suspended in a mixture of tetrahydrofurane (20 mL), water (10 mL) and triethylamine (0.97 g, 9.60 mmol, 1.34 mL, 3.0 eq.). Mixture of compound 67 and p-nitrophenol (1/1) (1.50 g, 6.40 mmol estimated, 2.0 eq. estimated) was solubilized in tetrahydrofurane (10 mL) and added dropwise to the ampicillin solution. The mixture was stirred for 2 h at RT. Solvents were evaporated under vacuum and the resulting aqueous solution was was acidified with (1N) hydrochloric acid solution. The precipitate was isolated by supernatant removal and was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), then 100/0.fwdarw.60/40 (17CV)] affording compound 68 (1.04 g, 78% yield) as a white powder.

c. Preparation of Compound 69

[0283] ##STR00104##

[0284] Intermediate 68 (600 mg, 1.40 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (1.0 g, 2.80 mmol, 2.0 eq.) were suspended in degassed dry tetrahydrofurane (70 mL) and stirred in the dark at RT for 18 h. Yellowish precipitate was collected by filtration and was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (5CV), 100/0.fwdarw.25/75 (20CV), then 25/75.fwdarw.25/75 (10CV)] followed by a direct lyophilization, affording compound 69 (156 mg, 14% yield). A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Phenomenex 10 μm PF-C18HP, gradient: water/acetonitrile (0.1% HCOOH) 82/18.fwdarw.82/18, 10 minutes and 82/18.fwdarw.0/100, 40 minutes) was done, followed by a direct lyophilization, affording compound 69 (41 mg, 4% yield) as a white solid (TMB24).

Example Part B: Synthesis Part II

Preparation of trialkylphosphine-gold-azide intermediates

a. Preparation of triethylphosphine-gold-azide (B)

[0285] ##STR00105##

[0286] Chloro(triethylphosphine) gold (I) A (5.00 g, 14.3 mmol, 1.00 eq.) and thallium(I)acethylacetonate (4.41 g, 14.5 mmol, 1.02 eq.) were suspended in dry toluene (330 mL). Reaction mixture was stirred in the dark at room temperature for 5 h. Reaction mixture was filtered through Celite® pad and washed with a minimum of toluene. Trimethylsilyl azide (2.30 g, 20.0 mmol, 2.65 mL, 1.40 eq.) was added dropwise to the filtrate, followed by dry methanol (40 mL). Reaction mixture was stirred in the dark at room temperature for 18 h. Precipitate was removed by filtration through Celite® pad and the filtrate was concentrated under vacuum. Residue was triturated in cold n-pentane. Supernatant was removed and the white precipitate washed 3 times with cold n-pentane and dried under vacuum, affording compound B (4.80 g, 94% yield) as a pale yellow solid.

b. Preparation of triisopropylphosphine-gold-azide (D)

[0287] ##STR00106##

[0288] Following the procedure described above, with chloro(triisopropylphosphine) gold (I) C (547 mg, 1.39 mmol, 1.00 eq.), thallium(I)acethylacetonate (431 mg, 1.42 mmol, 1.02 eq.) and trimethylsilyl azide (226 mg, 1.95 mmol, 0.26 mL, 1.40 eq.), afford compound D (497 mg, 89% yield) as a white solid.

c. Preparation of tricyclohexylphosphine-gold-azide (D)

[0289] ##STR00107##

[0290] Following the procedure described above, with chloro(tricyclohexylphosphine) gold (I) E (500 mg, 0.97 mmol, 1.00 eq.), thallium(I)acethylacetonate (302 mg, 0.99 mmol, 1.02 eq.) and trimethylsilyl azide (156 mg, 1.36 mmol, 0.18 mL, 1.40 eq.), afford compound F (460 mg, 91% yield) as a white solid.

1. Preparation of TMB25

[0291] ##STR00108##

a. Preparation of Intermediate 2

[0292] ##STR00109##

[0293] Methyl 4-ethynylcyclohexanecarboxylate 1 (250 mg, 1.50 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (10 mL) and water (1 mL). Lithium hydroxide hydrate (189 mg, 4.50 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting aqueous solution was acidified with (6N) hydrochloric acid solution. After evaporation under vacuum, the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.50/50 (12 CV)] affording compound 2 (80 mg, 35% yield) as a white powder.

b. Preparation of Compound 3

[0294] ##STR00110##

[0295] Compound 2 (80 mg, 0.52 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (241 mg, 0.68 mmol, 1.60 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 3 (110 mg, 41% yield) as a white solid (TMB25).

II. Preparation of TMB26

[0296] ##STR00111##

a. Preparation of Intermediate 6

[0297] ##STR00112##

[0298] 4-Hydroxy-D-phenylglycine 4 (1.00 g, 5.98 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (0.96 g, 23.92 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (0.78 g, 6.58 mmol, 0.64 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum affording compound 6 (770 mg, 52% yield) as a white solid, without further purification.

b. Preparation of Compound 7

[0299] ##STR00113##

[0300] Compound 6 (100 mg, 0.40 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (314 mg, 0.88 mmol, 2.2 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 7 (61 mg, 25% yield) as a white solid (TMB26).

III. Preparation of TMB27

[0301] ##STR00114##

a. Preparation of Intermediate 9

[0302] ##STR00115##

[0303] D-Phenylalanine 8 (1.00 g, 6.05 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (0.97 g, 24.20 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (0.79 g, 6.66 mmol, 0.65 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 9 (750 mg, 50% yield) as a colorless oil.

b. Preparation of Compound 10

[0304] ##STR00116##

[0305] Compound 9 (100 mg, 0.40 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (144 mg, 0.40 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 10 (110 mg, 45% yield) as a white solid (TMB27).

IV. Preparation of TMB28

[0306] ##STR00117##

a. Preparation of Intermediate 12

[0307] ##STR00118##

[0308] D-Proline 11 (1.00 g, 8.68 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (1.39 g, 34.72 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (1.13 g, 9.55 mmol, 0.93 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 12 (900 mg, 53% yield) as a colorless oil.

b. Preparation of Compound 13

[0309] ##STR00119##

[0310] Compound 12 (100 mg, 0.51 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (181 mg, 0.51 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 13 (120 mg, 43% yield) as a white solid (TMB28).

V. Preparation of TMB29

[0311] ##STR00120##

a. Preparation of Intermediate 15

[0312] ##STR00121##

[0313] D-Aspartic acid 14 (1.00 g, 7.51 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (1.20 g, 30.04 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (0.98 g, 8.26 mmol, 0.81 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 15 (640 mg, 40% yield) as a white powder.

b. Preparation of Compound 16

[0314] ##STR00122##

[0315] Compound 15 (200 mg, 0.93 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (365 mg, 1.02 mmol, 1.1 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 16 (167 mg, 31% yield) as a white solid (TMB29).

c. Preparation of TMB30

[0316] ##STR00123##

d. Preparation of Intermediate 18

[0317] ##STR00124##

[0318] D-Tryptophane 17 (1.00 g, 4.89 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (0.78 g, 19.56 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (0.64 g, 5.39 mmol, 0.52 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 18 (530 mg, 38% yield) as a white foam.

e. Preparation of Compound 19

[0319] ##STR00125##

[0320] Compound 18 (100 mg, 0.35 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (137 mg, 0.38 mmol, 1.1 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 19 (61 mg, 27% yield) as a white solid (TMB30).

VI. Preparation of TMB31

[0321] ##STR00126##

a. Preparation of Compound 21

[0322] ##STR00127##

[0323] 4-Ethynyl-1H-pyrazole 20 (100 mg, 1.09 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (426 mg, 1.09 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 21 (266 mg, 54% yield) as a white solid (TMB31).

VII. Preparation of TMB32

[0324] ##STR00128##

a. Preparation of Compound 23

[0325] ##STR00129##

[0326] 4-Prop-2-ynylmorpholine 22 (150 mg, 1.20 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (470 mg, 1.32 mmol, 1.1 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 23 (266 mg, 46% yield) as a white sticky paste. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 23 (123 mg, 21% yield) as a black sticky oil (TMB32).

VIII. Preparation of TMB33

[0327] ##STR00130##

a. Preparation of Compound 25

[0328] ##STR00131##

[0329] 1-Methyl-4-prop-2-ynyl-piperazine 24 (150 mg, 1.08 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (426 mg, 1.19 mmol, 1.1 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 25 (248 mg, 46% yield) as a yellow sticky paste. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 25 (74 mg, 14% yield) as a black sticky oil (TMB33).

IX. Preparation of TMB34

[0330] ##STR00132##

a. Preparation of Compound 27

[0331] ##STR00133##

[0332] Pent-4-yn-1-ol 26 (100 mg, 1.19 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (467 mg, 1.31 mmol, 1.1 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 27 (60% pure by LCMS). A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 27 (32 mg, 6% yield) as a colorless sticky oil (TMB34).

X. Preparation of TMB35

[0333] ##STR00134##

a. Preparation of Intermediate 29

[0334] ##STR00135##

[0335] N.sub.ω-Nitro-L-arginine 28 (1.00 g, 4.56 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (0.73 g, 18.24 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (0.60 g, 5.02 mmol, 0.49 mL, 1.1 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum affording compound 29 (600 mg, 44% yield) as a white powder.

b. Preparation of Compound 30

[0336] ##STR00136##

[0337] Compound 29 (100 mg, 0.33 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (119 mg, 0.33 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 30 (40 mg, 18% yield) as a white solid (TMB35).

XI. Preparation of TMB36

[0338] ##STR00137##

a. Preparation of Compound 32

[0339] ##STR00138##

[0340] But-3-yne-1-sulfonamide 31 (100 mg, 0.75 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (268 mg, 0.75 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 32 (165 mg, 45% yield) as a white solid (TMB36).

XII. Preparation of TMB37

[0341] ##STR00139##

a. Preparation of Intermediate 34

[0342] ##STR00140##

[0343] (R)-2-Amino-4-pentynoic acid, methyl ester 33 (100 mg, 0.79 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (281 mg, 0.79 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.50/50 (25 CV)] followed by a direct lyophilization, affording compound 34 in mixture with residual phosphine derivatives (240 mg) as a yellow oil. Mixture was used without further purification for the next step.

b. Preparation of Compound 35

[0344] ##STR00141##

[0345] Compound 34 in mixture with residual phosphine derivatives (120 mg, 0.25 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (10 mL) and water (2.5 mL). Lithium hydroxide hydrate (31 mg, 0.75 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting residue was purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes), followed by a direct lyophilization, to afford compound 35 (17 mg, 5% yield) as a white powder (TMB37).

XIII. Preparation of TMB38

[0346] ##STR00142##

a. Preparation of Intermediate 40

[0347] ##STR00143##

[0348] Pentynoic acid 36 (1.00 g, 10.19 mmol, 1.0 eq.) was solubilized in tetrahydrofurane (20 mL) and cooled down at −10° C. under argon atmosphere. Triethylamine (3.09 g, 30.57 mmol, 4.26 mL, 3.0 eq.) was added dropwise, followed by methylchloroformate 37 (1.06 g, 11.21 mmol, 0.87 mL, 1.1 eq.). Reaction mixture was stirred at −10° C. for 1 h. D-Serine methyl ester hydrochloride 39 (1.60 g, 10.19 mmol, 1.0 eq.) was added portionwise to the reaction mixture and the stirring was continued for 18 h at room temperature. Solvents were evaporated under vacuum, water (50 mL) was added to the residue and the resulting aqueous mixture was extracted with ethyl acetate (3×100 mL). Organic phase was dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (24CV)] affording compound 40 (306 mg, 15% yield) as a colorless oil.

b. Preparation of Intermediate 41

[0349] ##STR00144##

[0350] Compound 40 (100 mg, 0.50 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (179 mg, 0.50 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.30/70 (25 CV)] followed by a direct lyophilization, affording compound 41 (98 mg, 25% yield) as a white powder.

c. Preparation of Compound 42

[0351] ##STR00145##

[0352] Compound 41 (98 mg, 0.18 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (10 mL) and water (2.5 mL). Lithium hydroxide hydrate (22 mg, 0.53 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting residue was purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes), followed by a direct lyophilization, to afford compound 42 (16 mg, 17% yield) as a white powder (TMB38).

XIV. Preparation of TMB39

[0353] ##STR00146##

a. Preparation of Intermediate 45

[0354] ##STR00147##

[0355] Ethyl 6-bromonicotinic acid ester 43 (1.00 g, 4.35 mmol, 1.0 eq.), triethylamine (1.96 g, 19.60 mmol, 2.7 mL, 4.5 eq.) and copper iodide (41 mg, 0.22 mmol, 0.05 eq.) were solubilized/suspended in dry dichloromethane (40 mL) and degassed with argon bubbling. After trimethylsilyl acetylene 44 (0.55 g, 5.65 mmol, 0.78 mL, 1.3 eq.) and trans-dichlorobis(triphenylphosphine)palladium (II) (153 mg, 0.22 mmol, 0.05 eq.) addition, reaction vessel was sealed and reaction mixture was stirred at room temperature for 18 h. Reaction mixture was washed with a saturated solution of sodium chloride (2×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.80/20 (12CV)] affording compound 45 (892 mg, 83% yield) as a brown oil.

b. Preparation of Intermediate 46

[0356] ##STR00148##

[0357] Tetra-N-butylammonium fluoride (1.0M) solution in tetrahydrofurane (5.4 mL, 5.40 mmol, 1.5 eq.) was added dropwise to a solution of compound 45 (890 mg, 3.60 mmol, 1.0 eq.) in tetrahydrofurane (45 mL). Reaction mixture was stirred at room temperature for 30 min. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.75/25 (12CV)] affording compound 46 (138 mg, 22% yield) as a brown solid.

c. Preparation of Intermediate 47

[0358] ##STR00149##

[0359] Compound 46 (138 mg, 0.79 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (281 mg, 0.79 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.0/100 (25 CV)] followed by a direct lyophilization, affording compound 47 (236 mg, 56% yield) as a yellow solid.

d. Preparation of Compound 48

[0360] ##STR00150##

[0361] Compound 47 (220 mg, 0.41 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (20 mL) and water (5 mL). Lithium hydroxide hydrate (52 mg, 1.24 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); 100/0.fwdarw.0/100 (25 CV)] followed by a direct lyophilization, affording compound 48 (107 mg, 52% yield) as a yellowish solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 48 (76 mg, 37% yield) as pale yellow solid (TMB39).

XV. Preparation of TMB40

[0362] ##STR00151##

c. Preparation of Intermediate 50

[0363] ##STR00152##

[0364] D-Phenylglycine 49 (1.00 g, 6.61 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (1.06 g, 26.50 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (1.57 g, 13.22 mmol, 1.29 mL, 2.0 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12CV)] affording compound 50 (1.09 g, 71% yield) as a white solid.

d. Preparation of Compound 51

[0365] ##STR00153##

[0366] Compound 50 (100 mg, 0.43 mmol, 1.0 eq.) and tricyclohexylphosphine-gold-azide F (223 mg, 0.43 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: then 100/0-80/20 (30CV)] followed by a direct lyophilization, affording compound 51 (163 mg, 50% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 51 (67 mg, 21% yield) as a white solid (TMB40).

XVI. Preparation of TMB41

[0367] ##STR00154##

a. Preparation of Intermediate 53

[0368] ##STR00155##

[0369] Methyl ester D-phenylglycine hydrochloride 52 (2.06 g, 10.2 mmol, 1.0 eq.) was solubilized in tetrahydrofurane/water mixture (20 mL/10 mL). After triethylamine addition (3.12 g, 30.6 mmol, 4.3 mL, 3.0 eq.), propargylchloroformate 5 (1.22 g, 10.2 mmol, 1.0 mL, 1.0 eq.) was added dropwise and reaction mixture was stirred at room temperature for 2 h. Tetrahydrofurane was removed under vacuum, and the aqueous residue was extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.40/60 (12CV)] affording compound 53 (1.3 g, 51% yield) as a yellow oil.

b. Preparation of Intermediate 55

[0370] ##STR00156##

[0371] Compound 53 (1.00 g, 4.04 mmol, 1.0 eq.) was solubilized in toluene (20 mL). The solution was degassed by argon bubbling for 30 min. Benzyl azide 54 (1.07 g, 8.09 mmol, 1.01 mL, 2.0 eq.) was added to the reaction mixture and argon bubbling was continued for additional 10 min. Chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II) (77 mg, 0.20 mmol, 0.05 eq.) was added, vessel was sealed and reaction mixture was stirred at 80° C. for 18 h. Reaction was cooled down to room temperature and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (24 CV)] affording compound 55 (316 mg, 21% yield) as a brown/orange oil.

c. Preparation of Intermediate 56

[0372] ##STR00157##

[0373] Compound 55 (310 mg, 0.81 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (40 mL) and water (10 mL). Lithium hydroxide hydrate (102 mg, 2.44 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: CH.sub.2Cl.sub.2/MeOH; 100/0.fwdarw.80/20 (12 CV)], affording compound 56 (210 mg, 87% yield) as a white solid.

d. Preparation of Compound 57

[0374] ##STR00158##

[0375] Compound 56 (210 mg, 0.57 mmol, 1.0 eq.) was solubilized in methanol (30 mL) and was hydrogenated with Pd/C at 50° C. for 18 h (H.sub.2.fwdarw.5 bars). Reaction mixture was filtered through Celite® pad, washed with methanol and solvents were evaporated under vacuum. The residue was purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes), followed by a direct lyophilization, affording compound 57 (69 mg, 44% yield) as a white solid (TMB41).

XVII. Preparation of TMB42

[0376] ##STR00159##

a. Preparation of Intermediate 60

[0377] ##STR00160##

[0378] D-Biotin 58 (1.00 g, 4.09 mmol, 1.0 eq.), 1,3-dicyclohexylcarbodiimide (1.10 g, 5.32 mmol, 1.3 eq.) and N-hydroxysuccinimide 59 (518 mg, 4.50 mmol, 1.1 eq.) were suspended in dry dimethylformamide (30 mL) and stirred at room temperature for 18 h. Reaction mixture was filtered through cotton pad in order to remove urea byproduct derived from DCC. Solvents were evaporated under high vacuum. The residue was triturated in diethylether (100 mL) and collected by filtration through fritté affording compound 60 (1.65 g, yield >100%) as a white powder. The compound was used in the next step without any further purification.

b. Preparation of Intermediate 62

[0379] ##STR00161##

[0380] Compound 60 (850 mg, 2.49 mmol, 1.0 eq.) was solubilized in dry dimethylformamide (20 mL). After triethylamine (756 mg, 7.47 mmol, 1.0 mL, 3.0 eq.) and propargylamine 61 (274 mg, 4.98 mmol, 0.3 mL, 2.0 eq.) addition, reaction mixture was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.0/100 (12CV) and then 0/100.fwdarw.0/100 (3CV)]. A second elution was done [CH.sub.2Cl.sub.2/MeOH; gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.80/20 (12CV) and then 80/20.fwdarw.80/20 (3CV)], affording compound 62 (690 mg, yield 98%) as a light brown solid.

c. Preparation of Compound 63

[0381] ##STR00162##

[0382] Compound 62 (100 mg, 0.36 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (127 mg, 0.36 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.0/100 (25 CV)] followed by a direct lyophilization, affording compound 63 (71 mg, 31% yield) as a white solid. A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 63 (44 mg, 19% yield) as a white solid (TMB42).

XVIII. Preparation of TMB43

[0383] ##STR00163##

a. Preparation of Intermediate 65

[0384] ##STR00164##

[0385] 3,5-Dimethyl-4-iodoisoxazole 64 (5.00 g, 22.4 mmol, 1.0 eq.), triethylamine (9.07 g, 89.7 mmol, 12.5 mL, 4.0 eq.) and copper iodide (213 mg, 1.12 mmol, 0.05 eq.) were solubilized/suspended in dry dimethylformamide (50 mL) and degassed with argon bubbling. After trimethylsilyl acetylene 44 (2.62 g, 26.9 mmol, 3.7 mL, 1.2 eq.) dropwise addition, trans-dichlorobis(triphenylphosphine)palladium (II) (787 mg, 1.12 mmol, 0.05 eq.) was added portionwise and reaction vessel was sealed. Reaction mixture was stirred at 75° C. for 4 h. After cooling down at room temperature, reaction mixture was diluted in 200 mL of diethylether and washed with a saturated solution of sodium chloride (2×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 120 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.90/10 (12CV)] affording compound 65 (3.2 g, 74% yield) as a dark brown oil.

b. Preparation of Intermediate 66

[0386] ##STR00165##

[0387] Tetra-N-butylammonium fluoride (1.0M) solution in tetrahydrofurane (24.8 mL, 24.8 mmol, 1.5 eq.) was added dropwise to a solution of compound 65 (3.2 g, 16.60 mmol, 1.0 eq.) in tetrahydrofurane (100 mL). Reaction mixture was stirred at room temperature for 30 min. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 120 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.80/20 (12CV)] affording compound 66 (620 mg, 31% yield) as a white solid.

c. Preparation of Intermediate 47

[0388] ##STR00166##

[0389] Compound 66 (200 mg, 1.65 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (590 mg, 1.65 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (40 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.70/30 (25 CV)] followed by a direct lyophilization, affording compound 67 (100 mg, in mixture with residual phosphine derivatives). A second purification by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes) was done, followed by a direct lyophilization, affording compound 67 (20 mg, 3% yield) as a white solid (TMB43).

XIX. Preparation of TMB44

[0390] ##STR00167##

[0391] See procedure for the preparation of the intermediate 50

a. Preparation of Compound 68

[0392] ##STR00168##

[0393] Intermediate 50 (100 mg, 0.43 mmol, 1.0 eq.) and triisopropylphosphine-gold-azide D (171 mg, 0.43 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: then 100/0-50/50 (25 CV)] followed by a direct lyophilization, affording compound 68 (130 mg, 48% yield) as a white solid (TMB44).

XX. Preparation of TMB45

[0394] ##STR00169##

a. Preparation of Intermediate 70

[0395] ##STR00170##

[0396] L-Phenylglycine 69 (1.00 g, 6.61 mmol, 1.0 eq.) was suspended in water (10 mL). Solubilization occurs after sodium hydroxide (1.06 g, 26.5 mmol, 4.0 eq.) addition. Reaction mixture was cooled down at 0° C., and propargylchloroformate 5 (1.58 g, 13.2 mmol, 1.3 mL, 2.0 eq.) was added dropwise. Reaction mixture was allowed to warm up to room temperature for 30 minutes. Reaction mixture was acidified with (12N) hydrochloric acid solution, extracted with ethyl acetate (3×100 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (12 CV)] affording compound 70 (977 mg, 63% yield) as a white solid.

b. Preparation of Compound 71

[0397] ##STR00171##

[0398] Compound 70 (100 mg, 0.43 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (153 mg, 0.43 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.25/75 (25 CV)] followed by a direct lyophilization, affording compound 71 (114 mg, 45% yield) as a white solid (TMB45).

XXI. Preparation of TMB46

[0399] ##STR00172##

a. Preparation of Intermediate 72

[0400] ##STR00173##

[0401] Pentynoic acid 36 (1.00 g, 10.19 mmol, 1.0 eq.) was solubilized in tetrahydrofurane (20 mL) and cooled down at −10° C. under argon atmosphere. Triethylamine (3.09 g, 30.57 mmol, 4.26 mL, 3.0 eq.) was added dropwise, followed by methylchloroformate 37 (1.06 g, 11.21 mmol, 0.87 mL, 1.1 eq.). Reaction mixture was stirred at −10° C. for 1 h. Methyl ester D-phenylglycine hydrochloride 52 (2.10 g, 10.19 mmol, 1.0 eq.) was added portionwise to the reaction mixture and the stirring was continued for 18 h at room temperature. Solvents were evaporated under vacuum, water (50 mL) was added to the residue and the resulting aqueous mixture was extracted with ethyl acetate (3×100 mL). Organic phase was dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.40/60 (24CV)] affording compound 72 (1.40 g, 56% yield) as a colorless oil.

b. Preparation of Intermediate 73

[0402] ##STR00174##

[0403] Compound 72 (1.00 g, 4.08 mmol, 1.0 eq.) was solubilized in toluene (20 mL). The solution was degassed by argon bubbling for 30 min. Benzyl azide 54 (1.09 g, 8.15 mmol, 1.02 mL, 2.0 eq.) was added to the reaction mixture and argon bubbling was continued for additional 10 min. Chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II) (77 mg, 0.20 mmol, 0.05 eq.) was added, vessel was sealed and reaction mixture was stirred at 80° C. for 18 h. Reaction was cooled down to room temperature and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.0/100 (24 CV)] affording compound 73 (1.20 g, 79% yield) as a yellow foam.

c. Preparation of Intermediate 74

[0404] ##STR00175##

[0405] Compound 73 (1.20 g, 3.17 mmol, 1.0 eq.) was solubilized in a mixture of tetrahydrofurane (120 mL) and water (30 mL). Lithium hydroxide hydrate (400 mg, 9.51 mmol, 3.0 eq.) was added to the reaction mixture which was stirred at room temperature for 18 h. Solvents were evaporated under vacuum and the resulting residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: CH.sub.2Cl.sub.2/MeOH; 100/0.fwdarw.80/20 (12 CV)], affording compound 74 (1.10 mg, 99% yield) as a white foam.

d. Preparation of Compound 75

[0406] ##STR00176##

[0407] Compound 74 (1.10 g, 3.02 mmol, 1.0 eq.) was solubilized in methanol (60 mL) and was hydrogenated with Pd/C at 50° C. for 22 h (H.sub.2.fwdarw.5 bars), then at 50° C. for additional 24 h (H.sub.2.fwdarw.6 bars). Reaction mixture was filtered through Celite® pad, washed with methanol and solvents were evaporated under vacuum affording compound 75 (690 mg, 83% yield) as a white solid. A 200 mg portion of the residue was purified by semi-preparative HPLC (Gilson PLC 2020, column C8 Princeton SPHER.60-10 μm, gradient: water/acetonitrile (0.1% HCOOH) 95/5.fwdarw.95/5, 10 minutes and 95/5.fwdarw.0/100, 25 minutes), followed by a direct lyophilization, affording compound 75 (160 mg, calculated 67% yield) as a white solid (TMB46).

XXII. Sourcing of Intermediate TMB47

[0408] ##STR00177##

[0409] Ofloxacin 76 was sourced from Preswick Chemical Library®.

XXIII. Preparation of TMB48

[0410] ##STR00178##

a. Preparation of Intermediate 79

[0411] ##STR00179##

[0412] 5-(2-Oxo-hexahydro-thieno[3,4-d]imidazol-6-yl)-pentanoic acid methyl ester 77 (1.00 g, 3.87 mmol, 1.0 eq.), 4,4′-Dimethoxytrityl chloride 78 (3.93 g, 11.61 mmol, 3.0 eq.), triethylamine (467 mg, 4.64 mmol, 0.65 mL, 1.2 eq.) and DMAP (118 mg, 0.97 mmol, 0.25 eq.) were solubilized in dry pyridine (30 mL) and stirred at 75° C. for 18 h in a sealed reaction vessel. Reaction mixture was cooled down to room temperature and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 80 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV), 100/0.fwdarw.0/100 (12CV) and then 0/100.fwdarw.0/100 (3CV)], affording compound 79 (1.10 g, yield 51%) as a yellow foam.

b. Preparation of Intermediate 81

[0413] ##STR00180## ##STR00181##

[0414] Sodium hydride (60% in mineral oil) (196 mg, 4.90 mmol, 5.0 eq.) was suspended in dry dimethylformamide (20 mL) and cooled down to 0° C. under argon atmosphere. Compound 79 (550 mg, 0.98 mmol, 1.0 eq.) was solubilized in dry dimethylformamide (20 mL) and added dropwise to the sodium hydride suspension. Tetra-butylammonium iodide (72 mg, 0.20 mmol, 0.2 eq.) was added portionwise to reaction mixture, followed by propargyl bromide solution 80 (80% wt. in toluene) (140 mg, 1.18 mmol, 0.13 mL, 1.2 eq.). The mixture was stirred at room temperature for 3 h. After water addition (20 mL), volatiles were removed under vacuum. Residual aqueous phase was acidified (pH=2) by potassium bisulfate addition and extracted with ethyl acetate (3×100 mL). Organics were dried over magnesium sulfate and solvents were evaporated under vacuum affording compound 81 in mixture with compound 82 (75/25). The mixture was purified by flash chromatography [Biotage®; column AIT® 120 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (3 CV) and then 100/0.fwdarw.40/60 (22CV)]. A second elution was done [CH.sub.2Cl.sub.2/MeOH; gradient: 100/0.fwdarw.100/0 (3 CV), and then 100/0.fwdarw.75/25 (12CV)], affording compound 81 (180 mg, 31% yield) as an orange foam and a fraction of compound 81 in mixture with compound 82 (10/90) (320 mg, approx. 56% yield) as a black oil. Both fraction were combined for the next step.

c. Preparation of Intermediate 83

[0415] ##STR00182## ##STR00183##

[0416] A mixture of compound 81 and compound 82 (500 mg, approx. 0.85 mmol, 1.0 eq.), was solubilized in dichloromethane (50 mL) and, after trifluoroacetic acid addition (10 mL), stirred at room temperature for 1 h. Solvents were evaporated under high vacuum, affording a mixture of compound 83, compound 84 and byproduct issued of 4,4′-Dimethoxytrityl deprotection. This mixture was found to be not separable after biotage purification. As a consequence, the crude mixture was used for the next step (saponification).

d. Preparation of Intermediate 84

[0417] ##STR00184##

[0418] Previous crude mixture of compounds 83 and 84 was solubilized in a mixture of tetrahydrofurane (50 mL) and water (10 mL). Lithium hydroxide hydrate (409 mg, 9.77 mmol, estimated 5.0 eq.) was added to the reaction mixture which was stirred at room temperature for 3 h. Solvents were evaporated under vacuum and the resulting residue was acidified with HCl (2.0M) in diethyl ether. Solvent were removed under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0 (5 CV), 100/0.fwdarw.0/100 (15CV), then 0/100.fwdarw.0/100 (5CV)], followed by lyophilization, affording compound 84 (100 mg, calculated 72% yield for 3 steps) as a brown oil.

e. Preparation of Compound 85

[0419] ##STR00185##

[0420] Compound 84 (100 mg, 0.35 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (139 mg, 0.35 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (2 CV), then 100/0.fwdarw.50/50 (15 CV)] followed by a direct lyophilization, affording compound 85 (30 mg, 13% yield) as a white solid (TMB48).

XXIV. Preparation of TMB49

[0421] ##STR00186##

a. Preparation of Compound 87

[0422] ##STR00187##

[0423] Compound 86, (100 mg, 0.32 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (113 mg, 0.32 mmol, 1.0 eq.) were solubilized in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.100/0 (2 CV), then 100/0.fwdarw.40/60 (15 CV)] followed by a direct lyophilization, affording compound 87 (88 mg, 41% yield) as a pale yellow solid (TMB49).

XXV. Preparation of TMB50

[0424] ##STR00188##

a. Preparation of Compound 89

[0425] ##STR00189##

[0426] 1-Amino-3-butyne 88 (100 mg, 1.45 mmol, 0.12 mL, 1.0 eq.) and triethylphosphine-gold-azide B (517 mg, 1.45 mmol, 1.0 eq.) were solubilized in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.0/100 (15 CV)] followed by a direct lyophilization, affording compound 89 obtained as acetic acid salt (178 mg, 23% yield) as a pale yellow solid (TMB50).

XXVI. Preparation of Intermediate TMB51

[0427] ##STR00190##

[0428] Compound 86 nd was purified by flash chromatography (315 mg scale) [Biotage®; column AIT® 12 g; eluant: CH.sub.2Cl.sub.2/MeOH; gradient: 100/0.fwdarw.90/10 (12 CV)] followed by a lyophilization, affording compound 86 (185 mg, 59% yield) as a yellowish solid (TMB51).

XXVII. Preparation of TMB52

[0429] ##STR00191##

e. Preparation of Intermediate 88

[0430] ##STR00192##

[0431] p-Nitrophenyl-chloroformate 87 (3.7 g, 18.2 mmol, 1.0 eq.) was solubilized in tetrahydrofurane (12 mL) and added dropwise to a cold solution (0° C.) of propargylamine 61 (1.0 g, 18.2 mmol, 1.16 mL, 1.0 eq.) and triethylamine (3.7 g, 36.3 mmol, 4.90 mL, 2.0 eq.) in tetrahydrofurane (35 mL), under argon atmosphere. Reaction mixture was stirred for 3 h at room temperature and the precipitate was filtered off. The filtrate was evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 120 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.20/80 (15 CV)] affording compound 88 (2.0 g, 37% yield) as a pale yellow solid.

f. Preparation of Intermediate 89

[0432] ##STR00193##

[0433] A solution of compound 88 (1.00 g, 4.27 mmol, 1.0 eq.) in tetrahydrofurane (20 mL) was prepared. D-Phenylglycine 49 (0.64 g, 4.27 mmol, 1.0 eq.) was suspended in water (10 mL) and tetrahydrofurane (20 mL). Solubilization occurs after triethylamine (1.30 g, 12.81 mmol, 1.78 mL, 3.0 eq.) addition. This solution was added dropwise to the previous solution of compound 88. The reaction mixture was stirred at room temperature for 1 h and volatiles were evaporated under vacuum. The residual aqueous phase was cooled in an ice bath and acidified with with hydrochloric acid solution (12N). No precipitation occurs, but a gummy residue was formed. Supernatant was removed and the gummy residue was solubilized in ethyl acetate (20 mL), dried over magnesium sulfate and solvents were evaporated under vacuum. The residue was purified by flash chromatography [Biotage®; column AIT® 40 g; eluant: Cyclohexane/EtOAc; gradient: 100/0.fwdarw.100/0(3 CV), then 100/0.fwdarw.0/100 (15 CV)] affording compound 89 (240 mg, 24% yield) as a white solid.

g. Preparation of Compound 90

[0434] ##STR00194##

[0435] Compound 89 (100 mg, 0.43 mmol, 1.0 eq.) and triethylphosphine-gold-azide B (154 mg, 0.43 mmol, 1.0 eq.) were suspended in degassed dry tetrahydrofurane (20 mL) and stirred in the dark at room temperature for 18 h. Solvents were evaporated under vacuum and the residue was purified by flash chromatography [Biotage®; column AIT® 12 g; eluant: EtOAc/(EtOAc/H.sub.2O/AcOH 3/1/1); gradient: 100/0.fwdarw.20/80 (25 CV)] followed by a direct lyophilization, affording compound 90 (41 mg, 16% yield) as a white solid (TMB52).

Examples Part B: Cytotoxicity Studies

Materials & Methods:

[0436] The human hepatocyte cytotoxicity profile has been evaluated between 7 and 25 μM for TMB derivatives as a toxicity appeared from 25 μM for the initial hit TMB3 during a preliminary assessment.

[0437] The cytotoxicity profile depends on the nature of the chemical series. Also, apart from very short derivatives, the TMB compounds exhibit generally an acceptable cytotoxicity up to 17 μM allowing a therapeutic window with regard to the extremely weak minimum inhibitory concentrations obtained during microbiological tests and the extreme fragility of hepatocytes cells.

[0438] This preliminary in vitro cytotoxicity provides an indication of an acceptable therapeutic window if we consider that the MICs measured correspond to a range of concentrations of 0.08-0.63 μM for TMB3 compound depending on the bacteria strains and that a cytotoxicity appears at 20 μM for this molecule.

Cytotoxicity Profile

[0439] The cytotoxicity profile depends on the nature of the chemical series. Also, apart from very short derivatives, the compounds according to the invention exhibit generally an acceptable cytotoxicity up to 17 μM allowing a therapeutic window with regard to the extremely weak minimum inhibitory concentrations obtained during microbiological tests and the extreme fragility of hepatocytes cells.

Examples Part C: Biological Data

Materials & Methods

Antibacterial Profiles

[0440] The antibacterial profile of each derivative was evaluated by the standard microdilution method for susceptibility testing on 16 different strains, following the guidelines of CLSI. The selected panel included: [0441] S. aureus ATCC25923 (MecA negative, wild-type) [0442] S. aureus ATCC29213 (MecA negative, wild-type) [0443] S. aureus ATCC700699 (MecA positive, glycopeptide resistant) [0444] S. aureus ST20131365 (MecA positive, glycopeptide resistant) [0445] S. epidermidis ATCC14990 (MecA negative, wild-type) [0446] S. epidermidis ATCC35984 (MecA positive) [0447] S. epidermidis ST20140436 (MecA positive, glycopeptide resistant) [0448] S. epidermidis ST20150446 (MecA positive, glycopeptide resistant) [0449] E. faecalis JH2-2 (wild-type) [0450] E. faecalis UCN41 (VanA positive, glycopeptide resistant) [0451] E. faecalis V583 (VanB positive, glycopeptide resistant) [0452] E. faecium ATCC19434T (wild-type) [0453] E. faecium BM4147 (VanA positive, glycopeptide resistant) [0454] E. faecium AUS0004 (VanB positive, glycopeptide resistant) [0455] E. coli ATCC25922 (wild-type) [0456] Clostridium difficile ATCC9689 (wild-type)

Antibacterial Activity

[0457] Both series issued from the SAR study were evaluated for their antibacterial activity on a panel of characterized strains obtained from the French National Reference Centers for Staphylococci (Lyon, Pr. Fréd{tilde over (e)}rique Laurent) and for Enterococci (Caen, Pr. Vincent Cattoir), respectively. As shown in the following tables, both series contain very active compounds on multi-resistant Gram-positive bacteria (Minimal Inhibitory Concentrations lower than comparator antibiotic ampicillin). Furthermore, a very important antibacterial effect has been shown too on a strain of Clostridium difficile, a Gram-positive, spore-forming, bacillus responsible for post-antibiotic diarrhoae.

[0458] The antibacterial activity of test compounds was evaluated on a selected panel of Gram-positive (4 Staphylococcus aureus, 4 Staphylococcus epidermidis, 3 Enterococcus faecalis, 3 Enterococcus faecium, 1 Clostridium difficile) and Gram-negative strains (1 Escherichia coli, 1 Klebsiella pneumoniae, 1 Pseudomonas aeruginosa, 1 Acinetacter baumannii) with known features of antibiotic resistance (from wild-type susceptibility to multi-drug resistance). The Minimal Inhibitory Concentration (MIC) of test compounds was determined in duplicate by the microdilution method in 96-well microplates with a final volume of 100 μL per well and Mueller-Hinton broth as the growth medium, as recommended by the CLSI (Clinical and Laboratory Standards Institute, document Mo7-A10, USA).

Results:

[0459] Altogether, the data from Table 1, 2, 3, 4, 5, 6, 7, 9, 10, 11 and 12 obtained suggest that the compounds of the invention are excellent candidates for the development of new antibacterial agents against several multiresisant bacteria. In particular, as demonstrated in the following tables 1-12, examples of compounds of the present invention have Minimal Inhibitory Concentrations (MIC) close or lower than comparator antibiotic ampicillin and clearly demonstrating that they have antimicrobial activity on different strain or type of bacteria, in particular on multi-resistant bacteria.

[0460] In particular the compounds TMB1, TMB3, TMB4, TMB8, TMB12, TMB13, TMB14, TMB15, TMB18, TMB19, TMB20, TMB21, TMB22, TMB23, TMB24, TMB25, TMB26, TMB27, TMB28, TMB29, TMB30, TMB31, TMB32, TMB33, TMB34, TMB35, TMB36, TMB37, TMB38, TMB39, TMB42, TMB43, TMB45, TMB47, TMB48, TMB49, TMB50, TMB52 demonstrating that they have antimicrobial activity. In contrary the compounds TMB2, TMB5, TMB6, TMB7, TMB9, TMB41, TMB46, TMB51, which not comprise gold (I)-phosphine or 1,2,3-triazole moiety do not show an antibacterial activity.

In the tables below “Ampi” means ampicilline.

TABLE-US-00001 TABLE 1 Ampi TMB1 TMB3 TMB4 TMB5 S. aureus ATCC25923 0.5 0.125 0.125 0.125 2 ATCC29213 0.5 0.125 0.125 0.125 2 ATCC700699   8/>8 0.125/0.25  0.125/0.25  0.125 >8 ST20131365 4 0.125 0.125/0.25  0.125 >8 S. epidermis ATCC14990 1 ≤0.06 ≤0.06 ≤0.06 1 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 >8 ST20140436 4 ≤0.06 ≤0.06 ≤0.06   8/>8 ST20150446 8 ≤0.06 ≤0.06/0.125 0.125 >8 E. faecalis JH2-2 0.25/0.5  0.25/0.5  0.5/1   0.25/5   2/4 UCN41 0.5 0.5 1 0.5 8 V583 0.5 0.5/1   1 0.5 4 E. faecium ATCC19434T 0.5 0.5 1 0.5 8 BM4147 4/8 1 2 1 >8 AUS0004 >8 0.5 1 0.5 >8 E. coli ATCC25922 2 >8 >8 >8 >8 Clostridium ATCC70057 0.5 ≤0.06 ≤0.06-0.125 ≤0.06-0.125 4 difficile

TABLE-US-00002 TABLE 2 Ampi TMB3 TMB6 TMB7 TMB8 TMB9 S. aureus ATCC25923 0.5 0.125 0.5 1 0.25 1 ATCC29213 0.5 0.125 0.5/1   1 0.25 1 ATCC700699   8/>8 0.125/0.25  >8 >8 0.25/0.5  >8 ST20131365 4 0.125/0.25  >8 >8 0.25/0.5  >8 S. epidermis ATCC14990 1 ≤0.06 0.5 1 ≤0.06 1 ATCC35984 >8 ≤0.06 >8 >8 ≤0.06 >8 ST20140436 4 ≤0.06 4 8 0.125 8 ST20150446 8 ≤0.06/0.125 4/8 >8 0.125 8/>8 E. faecalis JH2-2 0.25/0.5  0.5/1   2/4 1/2 1 2 UCN41 0.5 1 8 4 1/2 8 V583 0.5 1 4 2 2 4 E. faecium ATCC19434T 0.5 1 8 8 1/2 8 BM4147 4/8 2 >8 >8 2 >8 AUS0004 >8 1 >8 >8 2 >8 E. coli ATCC25922 2 >8 >8 >8 >8 >8 Clostridium ATCC70057 0.5-1 ≤0.06-0.125 2-4 4 ≤0.06-0.125 2-4 difficile

TABLE-US-00003 TABLE 3 Ampi TMB18 TMB19 TMB20 TMB21 S. aureus ATCC25923 0.5 0.125 ≤0.06/0.125 0.125 ≤0.06/0.125 ATCC29213 0.5 0.125 0.125 ≤0.06/0.125 ≤0.06/0.125 ATCC700699   8/>8 0.125 0.125/0.25  0.125 0.125 ST20131365 4 0.125 0.125 0.125 0.125 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ST20140436 8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ST20150446 8 ≤0.06 ≤0.06 ≤0.06/0.125 ≤0.06 E. faecalis JH2-2 0.25/0.5  0.25 0.125/0.25  0.25 0.25 UCN41 0.5 0.25/0.5 0.125/0.25  0.25 0.25 V583 0.5 0.25/0.5 0.125/0.25  0.25/0.5  0.25/0.5  E. faecium ATCC19434T 0.5 0.25 0.125/0.25  0.25 0.25 BM4147 4 0.5 0.25 0.25/0.5  0.25/0.5  AUS0004 >8 0.25 0.125 0.25 0.25 E. coli ATCC25922 2 >8 >8 >8 >8 Clostridium ATCC700057 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 difficile

TABLE-US-00004 TABLE 4 Ampi TMB3 TMB12 TMB13 TMB14 TMB15 S. aureus ATCC25923 0.5 0.25 ≤0.06/0.125 0.125 0.125 0.25/0.5 ATCC29213 0.5 0.125 ≤0.06/0.125 0.125/0.25  0.125 0.5 ATCC700699   8/>8 0.125/0.25  0.125/0.25  0.25 0.125 0.5 ST20131365 4 0.25 0.5 0.25 0.125/0.25  0.25/0.5 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ST20140436 8 ≤0.06 ≤0.06 ≤0.06 ≤0.06/0.125 0.125 ST20150446 8 ≤0.06/0.125 ≤0.06 ≤0.06 ≤0.06/0.125 0.125/0.25 E. faecalis JH2-2 0.25/0.5  0.5/1 0.5 1/2 0.5 2 UCN41 0.5 1 0.5/1   1/2 0.5/1 2 V583 0.5 1 1 1/2 1 2/4 E. faecium ATCC19434T 0.5 0.5/1 0.5 0.5 0.5 2 BM4147 4 1 1 2 0.5 4 AUS0004 >8 1 0.5 1/2 0.5 2 E. coli ATCC25922 2 >8 >8 >8 >8 >8 Clostridium ATCC700057 0.5 0.125 ≤0.06 ≤0.06 0.125 0.125/0.25 difficile

TABLE-US-00005 TABLE 5 Ampi TMB3 TMB22 TMB23 TMB24 S. aureus ATCC25923 0.5 0.25 0.125 ≤0.06 0.125 ATCC29213 0.5 0.25 0.125 0.125 ≤0.06/0.125 ATCC700699 8 0.25 0.125/0.25  0.125 0.125/0.25  ST20131365 4 0.25 0.25 0.125 0.25 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06/0.125 ≤0.06/0.125 0.125 ST20140436 8 0.125 ≤0.06/0.125 ≤0.06/0.125 ≤0.06/0.125 ST20150446 4/8 0.125 ≤0.06/0.125 ≤0.06 ≤0.06/0.125 E. faecalis JH2-2 0.5 1 0.5 0.25 0.25/0.5  UCN41 0.5 1 1 0.125 0.25 V583 0.5 1 0.5 0.125/0.25  0.25/0.5  E. faecium ATCC19434T 0.5 1 1 0.25 0.5 BM4147 8 1 1 0.25/0.5  0.5 AUS0004 >8 1 1 0.25 0.5 E. coli ATCC25922 2 >8 >8 >8 >8 Clostridium ATCC700057 0.25/0.5 0.125 ≤0.06 ≤0.06 ≤0.06 difficile

TABLE-US-00006 TABLE 6 Ampi TMB3 TMB25 TMB27 TMB28 TMB29 TMB30 S. aureus ATCC25923 0.5 0.25 ≤0.06/0.125 0.25 0.125 0.125 0.25 ATCC29213 0.5 0.125 0.25 0.25/0.5  0.25 0.125 0.25 ATCC700699 8 0.25 0.25 0.25 0.125 0.125 0.25 ST20131365 4 0.25 ≤0.06/0.125 0.5 0.25/0.5 0.125 0.25 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ST20140436 8 0.125 ≤0.06/0.125 0.125 0.125 ≤0.06 0.125 ST20150446 8 0.125 0.125/0.25  0.125 0.125 ≤0.06/0125 0.125 E. faecalis JH2-2 0.5 0.5 0.125 0.5 0.25 0.25 0.5 UCN41 0.5 0.5 0.125 0.5/1   0.25 0.25 0.5 V583 0.5 1 0.125 1 0.25 0.25 0.5 E. faecium ATCC19434T 0.5 0.5 0.125 1 0.25 0.25 0.5 BM4147 8 1 0.125 1 0.5 0.25/0.5  1 AUS0004 >8 1 0.125 2 0.5 0.5 1 E. coli ATCC25922 2 >8 >8 >8 >8 >8 >8 Clostridium ATCC700057 0.5 0.06/0.125 ≤0.06 0.125 0.125/0.25 ≤0.06 0.125 difficile

TABLE-US-00007 TABLE 7 Ampi TMB3 TMB32 TMB33 TMB34 TMB37 S. aureus ATCC25923 0.5 0.25 ≤0.06/0.125 0.125 ≤0.06 0.125 ATCC29213 0.5 0.25 ≤0.06/0.125 0.25 ≤0.06 0.125/0.25 ATCC700699 >8 0.25 0.125/0.25  0.25 ≤0.06 0.25 ST20131365 4 0.25 0.125 0.25 ≤0.06 0.25 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ST20140436 4/8 0.125 ≤0.06/0125 0.25 ≤0.06 0.125 ST20150446 4/8 0.125 0.125/0.25  0.25 ≤0.06 0.125 E. faecalis JH2-2 0.5 0.5 0.125 0.25 0.125 0.5 UCN41 1 1 0.25 1 0.125 0.5 V583 0.5 1 0.25 0.5 0.125 0.5 E. faecium ATCC19434T 0.5 0.5 0.25 0.25 0.125 0.5 BM4147 4 1 0.25 0.5 0.125 0.5 AUS0004 >8 1 0.25 0.5 0.125 0.5 E. coli ATCC25922 2 >8 >8 >8 >8 >8 Clostridium ATCC700057 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 difficile

TABLE-US-00008 TABLE 8 Ampi TMB3 TMB26 TMB31 TMB35 TMB36 S. aureus ATCC25923 0.5 0.25 0.125 ≤0.06 0.125 ≤0.06 ATCC29213 0.5 0.25 0.125 ≤0.06 0.25 ≤0.06/0.125 ATCC700699 >8 0.25 0.25 0.125 0.25 0.125 ST20131365 4 0.25 0.125 ≤0.06/0.125 0.25 ≤0.06/0.125 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ≤0.06 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06/0.125 ≤0.06 ST20140436 4/8 0.125 ≤0.06 ≤0.06 0.125 ≤0.06 ST20150446 4/8 0.125 ≤0.06 ≤0.06 0.125/0.25  ≤0.06 E. faecalis JH2-2 0.5 0.5 0.25 0.25 1 0.25 UCN41 1 1 0.25 0.25 1 0.25 V583 0.5 1 0.5 0.25 1 0.5 E. faecium ATCC19434T 0.5 0.5 0.25 0.25 0.5 0.25 BM4147 4 1 0.5 0.25 1 0.5 AUS0004 >8 1 0.5 0.25 1 0.25 E. coli ATCC25922 2 >8 >8 >8 >8 >8 Clostridium ATCC700057 0.5 ≤0.06 0.125 ≤0.06 0.125 ≤0.06 difficile

TABLE-US-00009 TABLE 9 Ampi TMB3 TMB38 TMB39 TMB41 S. aureus ATCC25923 0.25 0.125 ≤0.06 ≤0.06 >8 ATCC29213 0.5 0.25 ≤0.06/0.125 ≤0.06 >8 ATCC700699 8 0.25 0.125 ≤0.06 >8 ST20131365 4 0.25 ≤0.06 ≤0.06 >8 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 >8 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 >8 ST20140436 4 0.25 ≤0.06 ≤0.06 >8 ST20150446 4 0.125 ≤0.06 ≤0.06 >8 E. faecalis JH2-2 0.5 0.5 0.125 0.25 >8 UCN41 0.5 0.5 0.125 0.25 >8 V583 0.5 1 0.125 0.25 >8 E. faecium ATCC19434T 0.5 0.5 ≤0.06 0.125 >8 BM4147 4 1 0.125 0.125 >8 AUS0004 >8 1 0.125 0.5 >8 Clostridium ATCC700057 0.5 0.125/0.25 0.125/0.25  0.25/0.5 >8 difficile

TABLE-US-00010 TABLE 10 Ampi TMB3 TMB42 TMB43 TMB45 TMB46 S. aureus ATCC25923 0.25 0.125 0.125 ≤0.06 0.125 >8 ATCC29213 0.5 0.25 0.25 0.125 0.25 >8 ATCC700699 8 0.25 0.25 0.125 0.25 >8 ST20131365 4 0.25 0.25 0.125 0.25 >8 S. epidermis ATCC14990 0.5 ≤0.06 ≤0.06 ≤0.06 ≤0.06 >8 ATCC35984 >8 ≤0.06 ≤0.06 ≤0.06 ≤0.06 >8 ST20140436 4 0.25 ≤0.06 0.125 ≤0.06 >8 ST20150446 4 0.125 ≤0.06 ≤0.06 ≤0.06 >8 E. faecalis JH2-2 0.5 0.5 0.25 0.25 0.5 >8 UCN41 0.5 0.5 1 0.25 0.25 >8 V583 0.5 1 1 0.25 0.5 >8 E. faecium ATCC19434T 0.5 0.5 0.25 0.125 0.25 >8 BM4147 4 1 0.5 0.125 0.25 >8 AUS0004 >8 1 0.5 0.25 0.5 >8 Clostridium ATCC700057 0.5 0.125/0.25 0.25 0.25 0.125 >8 difficile

TABLE-US-00011 TABLE 11 Ampi TMB3 TMB47 TMB48 TMB49 TMB50 S. aureus ATCC-25923 0.25 0.25 0.25 0.25 0.125 0.125 ATCC-700699 8 0.25 >8 0.5 0.25 0.125 S. epidermis ATCC-14990 0.5/1 ≤0.06 0.25 0.125 ≤0.06 ≤0.06 ATCC-35984 >8 ≤0.06 0.125 0.125 ≤0.06 ≤0.06 E. faecalis JH2-2 0.5 0.5 2 1 1 0.25 UCN41 0.5 0.5 2 1 1 0.5 E. faecium ATCC-19434T 0.5 0.5 4 1 0.5/1 0.25 BM-4147 4 1 1 2 1 0.25

TABLE-US-00012 TABLE 12 Ampi TMB3 TMB51 TMB52 Auranofin S. aureus ATCC-25923 0.25/0.5  0.25 2 ≤0.06/0.125 ≤0.06/0.125 ATCC-700699 8 0.25 >8 0.125 ≤0.06 S. epidermis ATCC-14990 0.5/1   ≤0.06 4 ≤0.06 ≤0.06 ATCC-35984 >8 ≤0.06 2 ≤0.06 ≤0.06 E. faecalis JH2-2 0.5 0.5 >8 0.25 ≤0.06/0.125 UCN41 0.5 0.5 >8 0.25 ≤0.06/0.125 E. faecium ATCC-19434T 0.5 0.5 >8 0.25 ≤0.06 BM-4147 4 1 >8 0.25 ≤0.06/0.125

CONCLUSION

[0461] The patent application described the synthesis and the biological properties of unprecedented organometallic species with impressive antibacterial properties and limited cytoxicities in vitro. These compounds were prepared using a copper free click chemistry reaction (1,3 dipolar cycloaddition) between a phosphine-gold(I) azide and an organic precursor bearing an alkyne moiety. In the resulting conjugate, the gold(I) atom has a phosphine ligand and is connected by a covalent gold-carbon bond to a 1,2,3-triazole ring (the three nitrogen are coming from the gold azide and the two carbons from the alkyne function). The antibacterial activity of gold(I) derivatives described in the patent application shows: (1) the crucial role of gold(I) in the biological properties, (2) an inverted correlation between the antibacterial activity and the size and/or lipophilicity of the phosphine ligand and (3) that the 1,2,3-triazole moiety alone is not responsible for biological activities reported in the present application.

REFERENCES

[0462] [1] Dalton Trans. 2014 Apr. 28; 43(16):5950-69. doi: 10.1039/c4dt00022f. [0463] [2] WO2017/093545. [0464] [3] Protective groups in organic synthesis”, T. W. Greene, P. G. M. Wuts, John Wiley and sons, Inc. [0465] [4] PAC, 1996, 68, 2193 (Basic terminology of stereochemistry (IUPAC Recommendations 1996)) on page 2205.