12-epi pleuromutilins

Abstract

A compound selected from 14-O-[((Alkyl-, cycloalkyl-, heterocycloalkyl-, heteoroaryl-, or aryl)-sulfanyl)-acetyl]-12-epi-mutilins, or 14-O-[((Alkyl-, cycloalkyl-, heterocycloalkyl-, heteoroaryl-, or aryl)-oxy)-acetyl]-12-epi-mutilins, wherein 12-epi-mutilin is characterized in that the mutilin ring at position 12 is substituted by two substituents, the first substituent at position 12 of the mutilin ring is a methyl group which methyl group has the inverse stereochemistry compared with the stereochemistry of the methyl group at position 12 of the naturally occurring pleuromutilin ring, the second substituent at position 12 of the mutilin ring is a hydrocarbon group comprising at least one nitrogen atom and all other substituents of the mutilin ring having the same stereochemistry compared with the stereochemistry of the substituents at the corresponding positions in the naturally occurring pleuromutilin ring; optionally in the form of a salt and/or solvate, wherein the naturally occurring pleuromutilin is of formula ##STR00001##
processes for the preparation of such compounds and their use as pharmaceuticals.

Claims

1. A compound selected from 14-O-[((Alkyl-, cycloalkyl-, heterocycloalkyl-, heteoroaryl-, or aryl)-sulfanyl)-acetyl]-12-epi-mutilins, or 14-O-[((Alkyl-, cycloalkyl-, heterocycloalkyl-, heteoroaryl-, or aryl)-oxy)-acetyl]-12-epi-mutilins, wherein 12-epi-mutilin is characterized in that the mutilin ring at position 12 is substituted by two substituents, the first substituent at position 12 of the mutilin ring is a methyl group which methyl group has the inverse stereochemistry compared with the stereochemistry of the methyl group at position 12 of the naturally occurring pleuromutilin ring, the second substituent at position 12 of the mutilin ring is a hydrocarbon group comprising at least one nitrogen atom, and all other substituents of the mutilin ring having the same stereochemistry compared with the stereochemistry of the substituents at the corresponding positions in the naturally occurring pleuromutilin ring; optionally in the form of a salt and/or solvate, wherein the naturally occurring pleuromutilin is of formula ##STR00366##

2. The compound according to claim 1 of formula ##STR00367## wherein the methyl group at position 12 of the mutilin ring has the inverse stereochemistry compared with the stereochemistry of the methyl group at position 12 of the naturally occurring pleuromutilin ring, all other substituents of the mutilin ring having the same stereochemistry compared with the stereochemistry of the substituents at the corresponding positions in the naturally occurring pleuromutilin ring, R.sub.1 is a hydrocarbon group comprising 1 to 16, in particular 1 to 12 carbon atoms comprising one N atom, optionally comprising one or more additional heteroatoms selected from N, O, S, halogen, in particular N, X is sulfur or oxygen, in particular sulfur, and R.sub.2 is a hydrocarbon group comprising 1 to 22 carbon atoms, optionally comprising heteroatoms selected from N, O, S, halogen, in particular N or O.

3. The compound according to claim 2, wherein R.sub.1 is either (C.sub.1-16)alkyl or (C.sub.2-16)alkenyl, substituted by heterocyclyl, including aliphatic heterocyclyl and aromatic heterocyclyl comprising 1 to 4 heteroatoms selected from N, O, S, with the proviso that at least one heteroatom is a nitrogen atom, or R.sub.1 is a group of formula ##STR00368## wherein YN(R.sub.3R.sub.4) is (C.sub.1-16)alkyl-N(R.sub.3R.sub.4), (C.sub.1-16)alkyl-(C.sub.6-14)aryl-N(R.sub.3R.sub.4), (C.sub.1-16)alkyl-(C.sub.6-14)aryl-(C.sub.1-16)alkyl-N(R.sub.3R.sub.4), (C.sub.1-16)alkyl-(C.sub.1-13)heterocyclyl N(R.sub.3R.sub.4), (C.sub.1-16)alkyl-(C.sub.1-13)heterocyclyl-(C.sub.1-16)alkyl-N(R.sub.3R.sub.4), carbonyl-N(R.sub.3R.sub.4), (C.sub.1-4)alkyl-carbonyl-N(R.sub.3R.sub.4), (C.sub.2-16)alkenyl-N(R.sub.3R.sub.4), (C.sub.2-16)alkenyl-(C.sub.6-14)aryl-N(R.sub.3R.sub.4), (C.sub.2-16)alkenyl-(C.sub.6-14)aryl-(C.sub.1-16)alkyl-N(R.sub.3R.sub.4), (C.sub.2-16)alkenyl-(C.sub.1-13)heterocyclyl-N(R.sub.3R.sub.4), (C.sub.2-16)alkenyl-(C.sub.1-13)heterocyclyl-(C.sub.1-16)alkyl-N(R.sub.3R.sub.4), wherein heterocyclyl includes aliphatic and aromatic heterocyclyl comprising at least one heteroatom selected from N, O, S and wherein alkyl, aryl, heterocyclyl or alkenyl is optionally substituted comprising substituents which optionally having heteroatoms selected from O, N, S, halogen; R.sub.3 and R.sub.4 independently of each other are hydrogen, (C.sub.1-16)alkyl, (C.sub.2-16)alkenyl, hydroxy(C.sub.1-16)alkyl, amino-(C.sub.1-16)alkyl, mono or di-(C.sub.1-6)alkylamino-(C.sub.1-16)alkyl, guanidino(C.sub.1-16)alkyl, ureido(C.sub.1-16)alkyl or thioureido(C.sub.1-16)alkyl, amino(C.sub.1-6)alkyl-(C.sub.6-14)aryl-(C.sub.1-6)alkyl, amino(C.sub.1-6)alkyl-(C.sub.6-14)aryl, guanidino(C.sub.1-6)alkyl-(C.sub.6-14)aryl-(C.sub.1-6)alkyl, amino-(C.sub.1-6)alkyloxy-(C.sub.1-6)alkyl, amino(C.sub.3-8)cycloalkyl, amino(C.sub.1-6)alkyl-(C.sub.3-8)cycloalkyl, amino(C.sub.3-8)cycloalkyl-(C.sub.1-6)alkyl, amino(C.sub.1-6)alkyl-(C.sub.3-8)cycloalkyl-(C.sub.1-6)alkyl, (C.sub.1-13)heterocyclyl-(C.sub.1-16)alkyl, (C.sub.6-14)aryl-(C.sub.1-6)alkyl, (C.sub.1-13)heterocyclyl, amino-(C.sub.6-14)aryl-(C.sub.1-16)alkyl, amino-(C.sub.1-6)alkyloxy-(C.sub.6-14)aryl-(C.sub.1-6)alkyl, amino(C.sub.1-6)alkyl-(C.sub.6-12)aryl-carbonyl, amino(C.sub.1-6)alkyl-amido-(C.sub.6-12)aryl(C.sub.1-6)alkyl, (C.sub.1-4)alkylcarbonyl, carbamimidoyl, carbamoyl, thiocarbamoyl, wherein heterocyclyl includes aliphatic and aromatic heterocyclyl comprising at least one heteroatom selected from N, O, S, and wherein alkyl, cycloalkyl, heterocyclyl, alkenyl or aryl is optionally further substituted, by amino(C.sub.1-4)alkyl, amido, mono or di-(C.sub.1-4)alkyl-amido, (C.sub.1-6)alkyloxy-carbonyl, halogen, oxo, hydroxy.

4. The compound according to claim 2, wherein R.sub.2 is (C.sub.1-16)alkyl, (C.sub.3-12)cycloalkyl, (C.sub.1-13)heterocyclyl, (C.sub.6-14)aryl, wherein heterocyclyl includes aliphatic and aromatic heterocyclyl comprising at least one heteroatom selected from N, O, S, and wherein alkyl, cycloalkyl, aryl, heterocyclyl is unsubstituted or substituted by substituents optionally having a heteroatom selected from O, N, S, and halogen.

5. The compound according to claim 4, wherein R.sub.2 is alkyl, optionally substituted by hydroxy or amino, (C.sub.3-12)cycloalkyl wherein the cycloalkyl group is optionally further substituted by amino or amino(C.sub.1-4)alkyl wherein the amino or aminoalkyl group is optionally further substituted by amino(C.sub.1-6)alkylcarbonyl and optionally (C.sub.1-4)alkyl, (C.sub.2-11)heterocyclyl, wherein a nitrogen in the ring as a heteroatom optionally is further substituted by amino(C.sub.1-6)alkylcarbonyl, cycloalkyl, optionally substituted by amino(C.sub.1-4)alkyl, wherein the amino group is optionally further substituted by amino(C.sub.1-6)alkylcarbonyl, hydroxy, amino, wherein the amino group is optionally further substituted by amino(C.sub.1-6)alkylcarbonyl and optionally (C.sub.1-4)alkyl, amino and hydroxy, wherein the amino group is optionally further substituted by amino(C.sub.1-6)alkylcarbonyl and optionally (C.sub.1-4)alkyl, (C.sub.1-4)alkylamino, wherein alkyl is optionally further substituted by one or more halogen atoms; aliphatic (C.sub.2-11)heterocyclyl, comprising 1 to 4 heteroatoms selected from N, O, S, wherein a nitrogen in the ring as heteroatom is optionally further substituted by (C.sub.1-4)alkyl, amino(C.sub.1-6)alkylcarbonyl, aryl, optionally substituted by hydroxy, halogen, amino, hydroxy(C.sub.1-4)alkyl, bis-(hydroxy(C.sub.1-4)alkyl), amino(C.sub.1-4)alkyl, bis-(amino(C.sub.1-4)alkyl), wherein the amino group in amino(C.sub.1-4)alkyl optionally is further substituted, aminocarbonyl, wherein the nitrogen optionally is substituted by amino(C.sub.1-12)alkyl, bis-(amino(C.sub.1-12)alkyl), hydroxy(C.sub.1-6)alkyl, bis-(hydroxy(C.sub.1-6)alkyl) or diamino(C.sub.1-6)alkyl, (C.sub.1-12)alkyl, which alkyl optionally is substituted by amino, which amino optionally is acylated, particularly amino is substituted by formyl, (C.sub.1-4)alkylcarbonyl, saturated or unsaturated heterocyclyl comprising 1 to 3 heteroatoms, particularly N, and 4 to 8, particularly 5 to 6 ring members (C.sub.6-14)aryl, particularly phenyl, which aryl optionally is substituted by amino(C.sub.1-4)alkyl, or the nitrogen of the aminocarbonyl group is part of (C.sub.3-8)heterocyclyl, including aliphatic and aromatic heterocyclyl, comprising one or more heteroatoms selected from N, O, S preferably N, wherein the heterocycle is optionally further substituted by amino(C.sub.1-4)alkyl; (C.sub.1-6)alkyl, which (C.sub.1-6)alkyl group is optionally substituted by aminocarbonyl, wherein the nitrogen of the aminocarbonyl group is optionally further substituted by amino(C.sub.1-12)alkyl, diamino-(C.sub.1-12)alkyl, bis-(amino(C.sub.1-12)alkyl), hydroxy(C.sub.1-6)alkyl, bis-(hydroxy(C.sub.1-6)alkyl), acylated amino(C.sub.1-4)alkyl, aromatic (C.sub.1-13)heterocyclyl, comprising 1 to 4 heteroatoms, wherein the aromatic heterocyclyl is optionally substituted by (C.sub.1-6)alkyl, amino or hydroxy wherein the alkyl group is optionally further substituted by halogen or amino or the aromatic heterocyclyl is optionally substituted by aminocarbonyl wherein the amino group is optionally further substituted by amino(C.sub.1-12)alkyl, bis-(amino(C.sub.1-12)alkyl), hydroxy(C.sub.1-6)alkyl, bis-(hydroxy(C.sub.1-6)alkyl) or diamino(C.sub.1-6)alkyl.

6. The compound according to claim 2, wherein R.sub.2 is amido-phenyl, amido(C.sub.1-4)alkyl-phenyl, wherein the nitrogen of the amido group is unsubstituted or substituted by amino(C.sub.1-8)alkyl, in which alkyl optionally is further substituted.

7. The compound according to claim 2, wherein R.sub.2 is amino(C.sub.3-12)cycloalkyl, amino(C.sub.1-4)alkyl(C.sub.3-12)cycloalkyl, amino(C.sub.3-12)cycloalkyl(C.sub.1-4)alkyl, or amino(C.sub.1-4)alkyl(C.sub.3-12)cycloalkyl(C.sub.1-4)alkyl, wherein the amino group is unsubstituted or substituted by amino(C.sub.1-6)alkylcarbonyl, or amino(C.sub.1-6)alkylcarbonyl and (C.sub.1-4)alkyl.

8. The compound according to claim 2, wherein R.sub.2 is (C.sub.2-11)heterocyclyl comprising 1 to 4 heteroatoms selected from N, O, S, wherein, if a nitrogen in the ring as a heteroatom is present, said nitrogen is unsubstituted or optionally further substituted by (C.sub.1-4)alkyl, amino(C.sub.1-6)alkylcarbonyl.

9. The compound according to claim 2, wherein X is S, and R.sup.2 is aminoethyl-amidomethyl-phenyl, aminopropyl-amidomethyl-phenyl, hydroxyphenyl-(amino)ethyl-amidomethyl-phenyl, aminomethyl-phenyl-(amino)ethyl-amidomethyl-phenyl, aminopropyl-amidophenyl, aminomethyl-phenylmethyl-amido-phenyl, aminomethyl-phenyl, aminoacetyl-aminomethyl-phenyl, bis(aminomethyl)phenyl, bisaminopropyl-amidomethyl-phenyl, (2-amino)-aminopropyl-amidomethyl-phenyl, aminoethyl-aminomethyl-phenyl, aminopropyl-aminomethyl-phenyl, allyl-aminomethyl-phenyl, aminomethyl-phenylmethyl-aminomethyl-phenyl, hydroxymethyl-phenyl, bis(hydroxymethyl)-phenyl, (tetrafluoro-hydroxymethyl)-phenyl, amino-hydroxy-cyclohexyl, hydroxyethyl, aminoethyl, piperazinocarbonyl-phenyl, aminomethyl-piperidine-carbonyl-phenyl, piperidine-ylmethyl-amido-phenyl, pyridine-ylmethyl-amido-phenyl, acetyl-aminopropyl-amido-phenyl, formyl-aminopropyl-amido-phenyl, amido-phenyl, aminohexyl-amidophenyl, aminoethyl-amidophenyl, (5-Amino)-4H-[1,2,4]triazol-3-yl, pyridinyl, hydroxyphenyl, fluorophenyl, purinyl, aminophenyl, acetyl-aminomethyl-phenyl, cyclopropyl-aminomethyl-phenyl, aminopropyl-amidopyridinyl, hydroxypropyl-amidophenyl, amino-purinyl, difluoroethylamino-cyclohexyl, amino-hydroxy-cyclohexyl, azepanyl, aminomethylcyclohexylmethyl, N-methyl-piperidinyl, piperidinyl, aminomethylcyclohexyl, aminopropylphenyl, phenyl, N-aminomethylcarbonyl-piperidinyl, N-aminoethylcarbonyl-piperidinyl, N-aminomethylcarbonyl-piperidinylmethyl, aminomethylamidomethylcyclohexyl, aminomethyl-pyridinyl, aminomethylamidocyclohexyl.

10. The compound according to claim 2, which is of formula ##STR00369## wherein n is 1 to 12 R.sub.3 is H, aminoethyl, aminopropyl, aminobutyl, aminopentyl, aminohexyl, aminooctyl, aminodecyl, dimethylaminopropyl, dimethylamidopentyl, guanidinobutyl, guanidinohexyl, carbamimidoyl, aminomethylcylcohexylmethyl, aminopropoxypropyl, aminocyclohexyl, hydroxyhexyl, dihydroxypropyl, aminomethylphenylmethyl, guanidinomethylphenylmethyl, phenylmethyl, morpholinopropyl, piperidinyl, hexyl, pyridinylethyl, allyl, amido-benzyl, aminopropyl-amidobenzyl, (2-amino)-amidoethyl-benzyl, (2-amino)-dimethylamidoethyl-benzyl, 2-amino-1-aminomethyl-ethyl, 5-amino-5-ethoxycarbonyl-pentyl, aminomethylphenylpropyl, aminomethylphenyl, aminophenymethyl, aminoethoxyphenylmethyl, aminomethyl-fluorophenyl-methyl, aminomethyl-difluorophenyl-methyl, and R.sub.4 is H(C.sub.1-4)alkylcarbonyl or aminomethylphenylcarbonyl.

11. The compound according to claim 2, wherein R.sub.1 is aminomethylphenylpropyl, aminoethylaminomethylphenylethenyl, aminoethylaminomethylphenyl ethyl, aminomethylphenylethyl, aminomethylphenylethyl, pyridinylethenyl, aminoethylamino-fluorophenyl-ethenyl.

12. The compound according to claim 1, selected from the group consisting of 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-ethylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-([Bis-(3-amino-propyl)-carbamoyl]-methyl})-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2,3-Diamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{4-[(2-Amino-ethylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(2-amino-ethylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(2-amino-ethylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-amino-butylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(5-amino-pentylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-guanidino-butylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(allylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-aminomethyl mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-guanidinomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(6-hydroxy-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(2,3-dihydroxypropylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-piperidylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-morpholin-4-yl-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-dimethylamino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(S)-5-amino-5-ethoxycarbonyl-pentylamino-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(4-Aminomethyl-benzylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(4-Aminomethyl-benzylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(4-Piperazinylcarbamoyl)-phenylsulfanyl]-acetyl})-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(4-Aminomethyl-piperidine-1-carbonyl)-phenylsulfanyl]-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(Piperidin-4-ylmethyl)-carbamoyl]-phenylsulfanyl)-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(Pyridin-4-ylmethyl)-carbamoyl]-phenylsulfanyl)-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[3-(3-Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl})-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Acetylamino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Formylamino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(3-Aminopropylcarbamoyl)-phenylsulfanyl)-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(3-Aminopropylcarbamoyl)-phenylsulfanyl)-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(8-amino-octylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(10-amino-decylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-Carbamoyl-phenylsulfanyl)-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-{[3-(3-amino-propoxy)-propylamino)]-methyl} mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12 [(2-pyridin-4-yl-ethyl amino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(6-Amino-hexylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(2-Amino-ethylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}-12-{[3-(4-aminomethyl-phenyl)-propylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-{[(4-Aminomethyl-cyclohexyl)-methylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-14-O-[(1-Methyl-piperidin-4-ylsulfanyl)-acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-14-O-[(Piperidin-4-ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl]mutilin, 12-epi-12-desvinyl-14-O-{[(4-Aminomethyl-cyclohexyl)-sulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-{[(3-amino-propyl)-acetylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-{(4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-(3-amino-propylcarbamoyl) mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-(4-aminomethyl-benzylcarbamoyl) mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[2-(3-amino-propylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3,5-Bis-hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(2,3,5,6-Tetrafluoro-4-hydroxymethyl)-phenylsulfanyl]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl]-mutilin, 12-epi-12-desvinyl-14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2-Hydroxy-ethylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2-Amino-ethylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(5-Amino-4H-1,2,4-triazol-3-yl)-sulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2-Amino-ethylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)-acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Hydroxy-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Fluoro-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(7H-Purin-6-yl)-sulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Amino-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-(Phenylsulfanyl-acetyl)-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Fluoro-phenylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Pyridin-2-ylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl)]-acetyl})-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(3-Amino-propionyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(3-Amino-propionyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-methylsulfanyl]-acetyl}-12-[(4-aminomethyl-phenylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-methylsulfanyl]-acetyl}-12-[(4-amino-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12{2-[4-(2-amino-ethoxy)-benzylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[{4-[(2-amino-ethoxy)-benzylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Aminomethyl-phenylsulfanyl)-acetyl]-12-[((4-aminomethyl-cyclohexyl)-methylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[(4-aminocyclohexyl)-amino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(4-carbamoylphenyl)-methylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(3-amino-propylcarbamoyl)-benzylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(5-dimethylcarbamoyl-pentyl amino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(2-amino-2-carbamoyl-ethyl)-benzylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(2-amino-2-dimethylcarbamoyl-ethyl)-benzylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-{[5-Aminomethyl-pyridin-2-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-Desvinyl-14-O-{[5-aminomethyl-pyridin-2-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-3-fluoro-benzyl amino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(4-Aminomethyl-cyclohexyl)-methylsulfanyl)-acetyl]{[(4-Aminomethyl-cyclohexyl)-methyl sulfanyl]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-methyl] mutilin, 12-epi-12-Desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl)}-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[5-Aminomethyl-pyridin-2-yl-sulfanyl]-acetyl}-12-[(4-aminomethyl-2,5-difluoro-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(2-amino-1-aminomethyl-ethylamino)-methyl] mutilin, 12-epi-12-Desvinyl-14-O-[(5-aminomethyl-pyridin-2-yl-sulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(2-Amino-acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(2-Amino-3-(4-hydroxy-phenyl)-propionylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(3-Amino-propionylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(2-Amino-acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[4-aminomethyl-benzylamino-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-[(2-Amino-acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-(6-amino-hexylamino-methyl) mutilin, 12-epi-12-desvinyl-14-O-{[(3-Acetylamino-methyl)-phenylsulfanyl]-acetyl}-12-[(3-amino-propyl amino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{(4-{[2-Amino-3-(4-aminomethyl-phenyl)-propionylamino]-methyl}-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl]mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{3-[(3-Amino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl-benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3-Allylaminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-{[3-(3-amino-propoxy)-propylamino]-methyl} mutilin, 12-epi-12-desvinyl-14-O-[(4-Cyclopropylaminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(4-Cyclopropylaminomethyl-phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl-benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl-benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl]-mutilin, 12-epi-12-desvinyl-14-O-[5-(3-Amino-propylcarbamoyl)-pyridin-2-ylsulfanyl]-acetyl-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2,5-Bis-aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(3,5-Bis-aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(2-guanidino-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[4-(3-Hydroxy-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2-Hydroxy-ethylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[3-(2,2-Difluoro-ethylamino)-cyclohexylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(2-Amino-7H-purin-6-ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(6-amino-octylamino)-methyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12-[(6-amino-hexylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-[5-Hydroxymethyl-pyridin-2-yl-sulfanylacetyl]-12-[(4-aminomethyl-3-fluoro-benzyl amino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{4-[(2-Amino-acetylamino)-cyclohexylsulfanyl]-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-[(5-Aminomethyl-pyridin-2-yl-sulfanyl)-acetyl]-12-[(4-aminomethyl-2,5-difluoro-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[(4-aminomethyl-2,5-difluoro-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12{2-[4-(2-amino-ethoxy)-benzylamino]-ethyl} mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(4-aminomethyl-phenylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)]-cyclohexylsulfanyl})-acetyl})-12-[(4-aminomethyl-phenylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-methylsulfanyl]-acetyl})-12-[(4-aminomethyl-phenylamino)-ethyl] mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl})-12-(8-amino-octyl) mutilin, 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl})-12-[3-(4-aminomethyl-phenyl)-propyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-yl-sulfanyl)-acetyl]-12-[3-(4-aminomethyl-phenyl)-propyl] mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-yl-sulfanyl)-acetyl]-12-(6-amino-hexyl) mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-yl-sulfanyl)-acetyl]-12-(8-amino-octyl) mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl})-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethenyl) mutilin, 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl})-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethyl) mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[2-(4-Aminomethyl-phenyl)-ethyl]-mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-((E)-2-pyridin-3-yl-ethenyl) mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-((E)-2-{4-[(2-Amino-ethylamino)-methyl]-phenyl}-ethenyl) mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethyl) mutilin, 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-((E)-2-{4-[(2-amino-ethylamino)-methyl]-3-fluoro-phenyl}-ethenyl) mutilin, 12-epi-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-((E)-2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethenyl) mutilin, 12-epi-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethyl) mutilin, 12-epi-12-desvinyl-14-O-[(5-Aminomethyl-pyridin-2-ylsulfanyl)-acetyl]-12-[2-(4-aminomethyl-benzoylamino)-ethyl] mutilin.

13. The compound according to claim 12, in the form of a salt and/or solvate.

14. The compound according to claim 1, optionally in the form of a pharmaceutically acceptable salt, for use as a pharmaceutical drug substance, in particular for use in the treatment of diseases mediated by Gram negative bacteria, in particular Escherichia coli.

15. A pharmaceutical composition comprising a compound according to claim 1 optionally in the form of a pharmaceutically acceptable salt, in association with at least one pharmaceutical excipient, optionally further comprising another pharmaceutically active agent.

16. The compound according to claim 12, optionally in the form of a pharmaceutically acceptable salt, for use as a pharmaceutical drug substance, in particular for use in the treatment of diseases mediated by Gram negative bacteria, in particular Escherichia coli.

17. A pharmaceutical composition comprising a compound according to claim 12, optionally in the form of a pharmaceutically acceptable salt, in association with at least one pharmaceutical excipient, optionally further comprising another pharmaceutically active agent.

Description

EXAMPLE 1

12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride

Step 1: Pleuromutilintosylate

(1) To a solution of 18.63 g of pleuromutilin and 9.39 g of toluenesulfonylchloride in 1400 mL of methylethylketone a solution of 4.98 g of triethylamine in 300 mL of methylethylketone is slowly added at rt. The mixture obtained is stirred for 24 h at rt, the formed precipitate is filtered off and to the filtrate obtained 2800 mL of water is added. The solution obtained is extracted 3 times with EtOAc and the organic phase obtained is dried over Na.sub.2SO.sub.4 and evaporated to dryness under reduced pressure. The crude product is used for the next step without further purification.

(2) .sup.1H-NMR (400 MHz, DMSO-d.sub.6, , ppm, characteristic signals): 0.49 (d, 3H, J=7 Hz, CH.sub.3-16); 0.8 (d, 3H, J=7 Hz, CH.sub.3-17); 1.02 (s, 3H, CH.sub.3-18), 1.29 (s, 3H, CH.sub.3-15); AB-system (v.sub.A=4.75, v.sub.B=4.62, J=50 Hz, CH.sub.2-22); 5.00 (m, 2H, H-20); 5.52 (d, 1H, J=8 Hz, H-14); 6.04 (dd, 1H, J=11, 18 Hz, H-19), 7.46 (d, 2H, J=8 Hz, arom.); 7.79 (d, 2H, J=8 Hz, arom.).

Step 2: 12-epi-Pleuromutilintosylate

(3) 20 g of Pleuromutilin tosylate was dissolved in 100 mL of THF and the solution obtained was heated to reflux. 20 mL of Diethylzinc (1M in heptane) was carefully added during 10 minutes and the reaction was kept at reflux for 7 hours. HPLC showed then a 46:53 ratio of Pleuromutilin tosylate:12-epi-Pleuromutilin tosylate by area. The batch was cooled to approx. 50 C., 2 mL of water was added; the resulting precipitate was filtered off and the organic phase was evaporated to dryness. The evaporation residue obtained was subjected to chromatography over silica gel using toluene/acetone 80:1 as eluent.

(4) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.79 (d, 2H, arom., J=8 Hz), 7.45 (d, 1H, arom., J=8 Hz), 5.9 (dd, 1H, H-19, J.sub.E=18 Hz, J.sub.Z=12 H.sub.Z), 5.49 (d, 1H, H-14, J=7.8 Hz), 4.92-4.85 (m, 2H, H-20), AB (2H, H-22, v.sub.A=4.76, v.sub.B=4.61, J=16 Hz), 4.25 (d, 1H, 11-OH, J=5.8 Hz), 2.40 (s, 3H, CH.sub.3), 1.31 (s, 3H, H-15), 0.99 (s, 3H, H-18), 0.80 (d, 3H, H-17, J=6.6 Hz), 0.52 (d, 3H, H-16, J=6.2 Hz).

(5) MS m/e: 550 [M.sup.++NH.sub.4].

Step 3: 12-epi-12-desvinyl-14-O-{(Toluene-4-sulfonyloxy)-acetyl}-12-formyl mutilin

(6) ##STR00032##

(7) 1 g of 12-epi-Pleuromutilin tosylate was dissolved in 25 mL of MeOH and 10 mL of EtOAc, cooled to 78 C. (carbon dioxide/acetone) and the mixture obtained was subjected to ozonisation at a flow rate of 70 L/h until a blue coloration persisted (ca. 10 minutes). To the still cold mixture was added 0.85 g of potassium iodide in 2.3 mL of water, 1.3 mL of acetic acid and 8 mL of MeOH in a manner that the internal temperature did not exceed 50 C. After addition the flask remained in the cooling bath and was left to warm up to about 0 C. At 0 C. 10 mL of 20% aqueous sodium thiosulfate solution was added to the mixture obtained and stirring was continued for 30 min. The reaction mixture was poured on 100 mL of water and extracted with 350 mL EtOAc. The combined organic layers obtained were washed with 5% NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The evaporation residue was subjected to chromatography over silica gel using cyclohexane/EtOAc 3:1 as eluent.

(8) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 9.59 (s, 1H, CHO), 7.80 (d, 2H, arom., J=8 Hz), 7.47 (d, 1H, arom., J=8 Hz), 5.34 (d, 1H, H-14, J=8.2 Hz), 4.83 (d, 1H, 11-OH, J=6.6 Hz), AB (2H, H-22, v.sub.A=4.76, v.sub.B=4.61, J=16 Hz), 3.57-3.5 (m, 1H, H-11), 2.41 (s, 3H, CH.sub.3), 1.30 (s, 3H, H-15), 1.07 (s, 3H, H-18), 0.93 (d, 3H, H-17, J=6.6 Hz), 0.50 (d, 3H, H-16, J=6.6 Hz).

(9) MS m/e: 552 [M.sup.++NH.sub.4].

(10) The ozonolysis of 11-Oxo pleuromutilin to give 12-desvinyl-12-formyl-11-oxo pleuromutilin using the same procedure is e.g. disclosed in: Tetrahedron, 37, 915 (1981).

Step 4: 12-epi-12-desvinyl-14-O-{(Toluene-4-sulfonyloxy)-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin of formula

(11) ##STR00033##

(12) 620 mg of 12-epi-12-desvinyl-14-O-{(toluene-4-sulfonyloxy)-acetyl}-12-formyl mutilin was dissolved in dichloroethane, 202 mg of N-BOC-diaminopropane dissolved in a minimum volume of dichloroethane was added and the mixture obtained was stirred at rt for one h. To the mixture obtained 541 mg of sodium trisacetoxyborohydride was added and the resulting slurry was stirred for another h at rt. HPLC then indicated the absence of the starting material. The mixture obtained was quenched with 5% aqueous NaHCO.sub.3 solution, the aqueous phase obtained was washed with DCM; the combined organic phases were dried over Na.sub.2SO.sub.4 and evaporated to dryness. The evaporation residue was subjected to chromatography over silica gel using EtOAc/triethylamine 100:1 as eluent.

(13) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.80 (d, 2H, arom., J=8 Hz), 7.47 (d, 1H, arom., J=8.2 Hz), 6.87 (bs, 1H, BOC-NH), 5.50 (d, 1H, H-14, J=7.2 Hz), AB (2H, H-22, v.sub.A=4.74, v.sub.B=4.62, J=16 Hz), 3.66 (bs, 1H, H-11), 2.41 (s, 3H, CH.sub.3), 1.36 (s, 9H, 3CH.sub.3), 1.30 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.80 (d, 3H, H-17, J=5.6 Hz), 0.51 (d, 3H, H-16, J=5.8 Hz).

(14) MS m/e: 693 [M.sup.++H].

Step 5: 12-epi-12-desvinyl-14-O-{{4-[(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(15) ##STR00034##

(16) 440 mg of tert-Butyl N-{3-[2-(4-mercapto-phenyl)-acetylamino]-propyl}-carbamate was dissolved in 20 mL of acetonitrile, 152 mg of potassium tert-butoxide was added followed by addition of 892 mg of 12-epi-12-desvinyl-14-O-[(toluene-4-sulfonyloxy)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin in one portion. The resulting slurry was stirred at rt for 1 h, diluted with 60 mL of water and extracted with DCM (4). The combined organic phases were washed 2 with aqueous 2N NaOH solution, water, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The evaporation residue obtained subjected to chromatography EtOAc/MeOH 8/1 to yield colourless crystals.

(17) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.03 (t, 1H, CONH, J=6 Hz), 7.27 (d. 2H. arom. J=8 Hz), 7.17 (d, 1H, arom., J=8.2 Hz), 6.83-6.74 (m, 2H, 2BOC-NH), 5.47 (d, 1H, H-14, J=7.2 Hz), 3.76 (s, 2H, H-22), 3.40 (bs, 1H, H-11), 1.36 (s, 9H, 3CH.sub.3), 1.30 (s, 3H, H-15), 0.80-0.76 (m, 6H, H-18, H-17), 0.57 (d, 3H, H-16, J=5.6 Hz).

Step 6: 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride

(18) ##STR00035##

(19) 654 mg of 12-epi-12-desvinyl-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin was dissolved in 2 mL of DCM, 11.5 mL of 1N HCl in diethyl ether was added and the mixture obtained was stirred at rt for 1 h. A precipitate formed and was collected with suction, washed 5 with diethyl ether and dried in a high vacuum overnight. The title compound was obtained in the form of a trihydrochloride.

(20) NMR: .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.29 (d, 2H, arom., J=8.4 Hz), 7.21 (d, 2H, arom., J=8.4 Hz), 5.44 (d, 1H, H-14, J=6 Hz), 5.36 (d, 11-OH, J=4 Hz), 1.35 (s, 3H, H-15), 0.57 (d, 3H, H-16, J=5.2 Hz).

(21) MS m/e: 645 [M.sup.++H].

Preparation of step 5Intermediate tert-Butyl N-[3-[[2-(4-Sulfanylphenyl)-acetyl]-amino]-propyl]-carbamate

Step a: 2-(4-Acetylsulfanylphenyl)-acetic acid

(22) 605 mg of 4-mercaptophenylacetic acid and 1.44 g of diisopropylethylamine was dissolved in 10 mL of DCM under an argon atmosphere, and the mixture obtained was cooled in an ice bath and treated with 532 mg of acetic anhydride. The mixture obtained was stirred 10 min with cooling and then 45 min at rt. To the mixture obtained 10 mL of 1N HCl was added and vigorous stirring was maintained for 10 min, the phases obtained were separated, the aqueous phase obtained was washed once with DCM and the combined organic phases were dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound was obtained in the form of pale yellow crystals (containing residual solvent).

Step b: S-[4-[2-[3-(tert-Butoxycarbonylamino)-propylamino]-2-oxo-ethyl]-phenyl]-ethanethioate

(23) 770 mg of 2-(4-acetylsulfanylphenyl)-acetic acid, 638 mg of N-BOC-1,3-diaminopropane and 560 mg of HOBt was dissolved in 15 mL DCM, stirred for 20 min at rt, cooled in an ice bath and treated with 755 mg of DCC. The cooling bath was removed and resulting slurry was stirred at rt for 1 hour and filtered from dicyclohexylurea. The filtrate obtained was washed with aqueous 10% K.sub.2CO.sub.3, 0.1N HCl and 5% NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The evaporation residue obtained was triturated with 10 mL of acetonitrile, filtered from insoluble material and brought to dryness. The title compound in the form of pale yellow soft crystals was obtained.

Step c: tert-Butyl N-[3-[[2-(4-sulfanylphenyl)-acetyl]-amino]-propyl]-carbamate

(24) 560 mg (1.53 mmol) of S-[4-[2-[3-(tert-butoxycarbonylamino)-propylamino]-2-oxo-ethyl]-phenyl]-ethanethioate was dissolved in 10 mL of MeOH, 422 mg (3.06 mmol) of potassium carbonate in 4 mL of water was added and the resulting yellow solution was stirred at ambient temperature for 1 hour, followed by dilution with 40 mL of water plus 7 mL of 1N HCl and extraction with 10 mL of dichloromethane. The aqueous phase was washed with dichloromethane (2), the combined organic phases washed with 5% NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound was obtained in the form of pale yellow crystals which were used in the next step without further purification.

(25) According, e.g. analogously, to a method as conventional, or according, e.g. analogously to a method as set out under Example 1 above, but using appropriate starting materials the compounds of formula

(26) ##STR00036##

(27) wherein R.sub.1EX and R.sub.2EX are as defined in Table 1 are prepared. Chemical characterisation data are also set out in Table 1.

(28) TABLE-US-00001 TABLE 1 Example R.sub.2EX R.sub.1EX 2 12-epi-12-desvinyl-14-O-{{4-[(2-Amino- ethylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 7.29 (d, 2H, arom., J = 7.2 Hz), 7.23 (d, 2H, arom., J = 9 Hz), 5.44 (d, 1H, H-14, J = 8 Hz), 3.81 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.84 (s, 3H, H-18) 0.58 (d, 3H, H-16, J = 4.8 Hz) MS m/e: 631 [M.sup.+ + H] 3 12-epi-12-desvinyl-14-O-{{4-([Bis-(3-amino- propyl)-carbamoyl]-methyl}-phenylsulfanyl)- acetyl}-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride MS m/e: 702 [M.sup.+ + H] 0 4 12-epi-12-desvinyl-14-O-{{4-[(2,3-Diamino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride MS m/e: 694 [M.sup. + Cl] The required [2-Amino-1-(tert- butoxycarbonylamino-methyl)-ethyl]-carbamic acid tert-butyl ester is e.g. described in U.S. Pat. No. 4,933,470 5 12-epi-12-desvinyl-14-O-{{4-[(2-Amino- ethylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(2-amino-ethylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 7.29 (d, 2H, arom., J = 8 Hz), 7.22 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H-14, J = 5.6 Hz), 3.80 (s, 2H H-22), 1.36 (s, 3H, H-15), 0.82 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.57 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 617 [M.sup.+ + H]. 6 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(2-amino-ethylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 7.29 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8.2 Hz), 5.44 (d, 1H, H-14, J = 5.8 Hz), 1.36 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.4 Hz), 0.57 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 631 [M.sup.+ + H] 7 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(4-amino-butylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 8.45 (t, 1H, CONH, J = 6 Hz), 7.99 (bs, 6 H, NH.sub.3.sup.+), 7.28 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H-14, J = 6.4 Hz), 1.34 (s, 3H, H-15), 0.85- 0.79 (m, 6H, H-17, H-18), 0.57 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 659 [M.sup.+ + H] 8 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(5-amino-pentylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 8.41 (t, 1H, CONH, J = 6 Hz), 8.0 (bs, 6 H, NH.sub.3.sup.+), 7.29 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.42 (bs, 1H, H-14), 1.35 (s, 3H, H-15), 0.85-0.79 (m, 6H, H-17, H-18), 0.57 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 673 [M.sup.+ + H] 0 9 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(4-aminomethyl-benzylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 7.68 (d, 2H, arom., J = 8 Hz), 7.55 (d, 2H, arom., J = 8.2 Hz), 7.26 (d, 2H, arom., J = 8 Hz), 7.18 d, 2H, arom., J = 8.2 Hz), 5.42 (d, 1H, H-14, J = 7.2 Hz), 1.34 (s, 3H, H- 15), 0.56 (d, 3H, H-16, J = 5.6 Hz) MS m/e 707 [M.sup.+ + H] 10 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.28 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H-14, J = 7.2 Hz), 3.8 (s, 2H, H-22), 1.35 (s, 3H, H- 15), 0.57 (d, 3H, H-16, J = 4.6 Hz) MS m/e: 687 [M.sup.+ + H] 11 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.28 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H-14, J = 7 Hz), 1.35 (s, 3H, H-15), 0.85 (s, 3H, H- 18), 0.80 (d, 3H, H-17, J = 6.4 Hz), 0.57 (d, 3H, H- 16, J = 5.4 Hz) MS m/e: 730 [M.sup.+ + H] The required tert-Butyl (NE)-N-[(6- aminohexylamino)-(tert-butoxycarbonylamino)- methylene]-carbamate was prepared according to: Journal of Medicinal Chemistry, 44(18), 2950; 2001 12 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(4-guanidino-butylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.32 (d, 2H, arom., J = 8 Hz), 7.25 (d, 2H, arom., J = 8 Hz), 5.43 (d, 1H, H-14, J = 7.4 Hz), 4.44 (s, 2H, PhCH.sub.2), 3.87 (s, 2H H-22), 1.35 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.57 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 702 [M.sup.+ + 2H] 13 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(allylamino)-methyl] mutilin dihydrochloride MS m/e: 672 [M + formate] 0

EXAMPLE 14

12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-aminomethyl mutilin dihydrochloride

(29) ##STR00061##

Step 1: 12-cpi-12-desvinyl-14-O-{{4-[(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methylphenylsulfanyl]}-acetyl}-12-aminomethyl mutilin

(30) 120 mg of N,N-Dimethylbarbituric acid and 6 mg of tetrakis(triphenylphosphine) palladium (0) was placed under a positive argon stream in a three necked round bottom flask together with a stirrer. 187 mg (0.257 mmol) of 12-epi-12-desvinyl-14-O-{{4-[(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methylphenylsulfanyl]}-acetyl}-12-[(allylamino)methyl]mutilin (compound of Example 13 prior to BOC-deprotection) was dissolved in 5 mL of DCM and degassed in a ultrasonic bath for 15 min and then added to the reaction flask via syringe and septum. The mixture was stirred for 6 hours under an inert atmosphere. HPLC indicated that there was still starting material left, hence 120 mg of N,N-Dimethylbarbituric acid and 6 mg of tetrakis(triphenylphosphine) palladium (0) were added to the mixture obtained and stirring was continued overnight until the reaction was deemed complete. The mixture obtained was diluted with DCM and washed with 10% K.sub.2CO.sub.3 (2), the combined aqueous phases were washed with DCM and the combined organic layers obtained were dried over Na.sub.2SO.sub.4 and evaporated to dryness. An orange residue was obtained which was subjected to column chromatography (EtOAc/Et.sub.3N 50:1 and EtOH). The title compound was obtained in the form of orange-red crystals.

Step 2: 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-aminomethyl mutilin dihydrochloride

(31) 100 mg of 12-epi-12-desvinyl-14-O-{{4-[(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methylphenylsulfanyl]}-acetyl}-12-aminomethyl mutilin was dissolved in 0.5 mL of DCM, treated with 5 mL of 1N HCl in diethyl ether and stirred at rt for 1 h. A precipitate formed and was collected with suction filtration, washed 5 with diethyl ether, dried 1 h at a rotary evaporator and purified via reverse chromatography with acetonitrile/H.sub.2O (0-75%). The according fractions were identified by HPLC, freed from organic solvent and lyophilized overnight. The title compound was obtained in the form of colourless crystals.

(32) MS m/e: 588 [M.sup.++H].

(33) According, e.g. analogously, to a method as conventional, or according, e.g. analogously to a method as set out under Example 1 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 2 are prepared. Chemical characterisation data are also set out in Table 2.

(34) TABLE-US-00002 TABLE 2 Example R.sub.2EX R.sub.1EX 15 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(benzylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.29 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.41 (d, 1H, H- 14, J = 7 Hz), 3.78 (s, 2H H-22), 1.34 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.56 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 678 [M.sup.+ + H] 16 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(4-guanidinomethyl-benzylamino)- methyl] mutilin trihydrochloride MS m/e: 750 [M.sup.+ + H] The required tert-Butyl (NE)-N-[[[4- (aminomethyl)phenyl]-methylamino]-(tert- butoxycarbonylamino)-methylene]-carbamate was prepared in analogy to: J. Med. Chem, 44(18), 2950; 2001 17 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(6-hydroxy-hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.29 (d, 2H, arom., J = 8 Hz), 7.20 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H- 14, J = 7 Hz), 3.80 (s, 2H H-22), 1.35 (s, 3H, H-15), 0.85 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.2 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 688 [M.sup.+ + H] 18 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(2,3-dihydroxypropylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.28 (d, 2H, arom., J = 8 Hz), 7.18 (d, 2H, arom., J = 8 Hz), 3.80 (s, 2H, H- 22), 1.33 (s, 3H, H-15), 0.80 (d, 3H, H-17, J = 5 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 662 [M.sup.+ + H] (2,2-Dimethyl-[1,3]dioxolan-4-yl)-methylamine was used as intermediate for step 4 19 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(4-piperidylamino)-methyl] mutilin trihydrochloride MS m/e: 671 [M.sup.+ + H] 0 20 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(3-morpholin-4-yl-propylamino)- methyl] mutilin trihydrochloride MS m/e: 715 [M.sup.+ + H] 21 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(3-dimethylamino-propylamino)- methyl] mutilin trihydrochloride MS m/e: 673 [M.sup.+ + H] 22 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylcarbamoyl)-methyl]-phenylsulfanyl}- acetyl}-12-[(S)-5-amino-5-ethoxycarbonyl- pentylamino-methyl] mutilin trihydrochloride MS m/e: 745 [M.sup.+ + H] The necessary Lysine-(N.sup.2BOC)-ethyl ester was prepared from N.sup.2BOC-Lysine using EtOH/1.1 eq of sulfuric acid

EXAMPLE 23

12-epi-12-desvinyl-14-O-{[4-(4-Aminomethyl-benzylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(35) ##STR00078##

(36) was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(37) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.86 (d, 2H, arom., J=8 Hz), 7.44-7.31 (m, 6H, arom.), 5.46 (d, 1H, H-14, J=6.6 Hz), 5.36 (d, 1H, 11-OH, J=5.6 Hz), 4.44 (d, 2H, Ph-CH.sub.2CO, J=5.2 Hz), 3.97 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.80 (d, 3H, H-17, J=6.2 Hz), 0.59 (d, 3H, H-16, J=5.6 Hz).

(38) MS m/e: 735 [M.sup.++H].

Preparation of the intermediate Thioacetic acid S-{4-[4-(tert-butoxycarbonylamino-methyl)-benzylcarbamoyl]-phenyl} ester

Step a: Thioacetic acid S-(4-chlorocarbonyl-phenyl) ester

(39) 4.48 g of 4-acetylsulfanylbenzoic acid was dissolved in 50 mL of DCM, treated with 5.8 mL of oxalyl chloride and one drop of DMF and the mixture obtained was stirred at rt for 90 min. The volatiles were distilled off in vacuo leaving the title compound as residue,

Step b: Thioacetic acid S-{4-[4-(tert-butoxycarbonylamino-methyl)-benzylcarbamoyl]-phenyl} ester

(40) 820 mg of tert-butyl N-[[4-(aminomethyl)-phenyl]-methyl]-carbamate and 421 mg of TEA was dissolved in 10 mL of DCM, cooled in an ice bath and 725 mg of thioacetic acid S-(4-chlorocarbonyl-phenyl) ester, dissolved in 5 mL of DCM, was slowly added. The mixture obtained was stirred for 30 min at rt, the phases were separated, the organic phase obtained was washed successively with 2N HCl and 5% aqueous NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound was obtained in the form of pale brown crystals.

(41) According, e.g. analogously, to a method as conventional, or according, e.g. analogously to a method as set out under Examples 1 and 23 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 3 are prepared. Chemical characterisation data are also set out in Table 3.

(42) TABLE-US-00003 TABLE 3 Example R.sub.2EX R.sub.2EX 24 12-epi-12-desvinyl-14-O-{[4-(4- Aminomethylbenzylcarbamoyl)-phenylsulfanyl]- acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 9.23 (t, 1H, NH, J = 5.2 Hz), 7.86 (d, 2H, arom., J = 8 Hz), 7.44-7.24 (m, 6H, arom.), 5.46 (d, 1H, H-14, J = 6.8 Hz), 1.36 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.78 (d, 3H, H-17, J = 6 Hz), 0.58 (d, 3H, H-16, J = 5 Hz) MS m/e: 777 [M.sup.+ + H] 0 25 12-epi-12-desvinyl-14-O-{[(4- Piperazinylcarbamoyl)-phenylsulfanyl]-acetyl}-12- [(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride MS m/e: 727 [M.sup.+ + H] 26 12-epi-12-desvinyl-14-O-{[4-(4-Aminomethyl- piperidine-1-carbonyl)-phenylsulfanyl]-acetyl}-12- [(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.40 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H- 14, J = 7 Hz), 3.93 (s, 2H, H-22), 1.34 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.4 Hz), 0.55 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 755 [M++ H] 27 12-epi-12-desvinyl-14-O-{(4-[(Piperidin-4- ylmethyl)-carbamoyl]-phenylsulfanyl)-acetyl}-12- [(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.82 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 4.95 (d, 1H, H- 14, J = 6.6 Hz), 5.37 (d, 1H, 11-OH, J = 5.4 Hz), 1.35 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.58 (d, 3H, H-16, J = 5.5 Hz) MS m/e: 755 [M.sup.+ + H] 28 12-epi-12-desvinyl-14-O-{(4-[(Pyridin-4-ylmethyl)- carbamoyl]-phenylsulfanyl)-acetyl}-12-[(6-amino- hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 9.53 (bs, 1H, CONH), 7.99-7.89 (m, 6H, arom.), 7.42 (d, 1H, arom., J = 8 Hz), 5.47 (d, 1H, H-14, J = 6.8 Hz), 3.98 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.90 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6 Hz), 0.59 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 707 [M.sup.+ + H] 29 12-epi-12-desvinyl-14-O-{[3-(3- Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}- 12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.70 (d, 1H, arom., J = 8 Hz), 7.50-7.37 (m, 3H, arom.), 5.45 (d, 1H, H- 14, J = 6.8 Hz), 5.34 (bs, 1H, 11-OH), 3.93 (s, 2H, H- 22), 3.72 (bs, 1H, H-11), 1.33 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.2 Hz), 0.56 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 715 [M.sup.+ + H] 0 30 12-epi-12-desvinyl-14-O-{[4-(3-Acetylamino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3- amino-propylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.81 (d, 2H, arom., J = 8 Hz), 7.39 (d, 2H, arom., J = 8 Hz), 5.47 (d, 1H, H- 14, J = 8 Hz), 5.34 (bs, 1H, 11-OH), 3.96 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.81 (d, 3H, H- 17, J = 5.8 Hz), 0.59 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 673 [M.sup.+ + H] 31 12-epi-12-desvinyl-14-O-{[4-(3-Formylamino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3- amino-propylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.82 (bs, 1H, CONH), 7.81 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.46 (d, H, H-14, J = 6.8 Hz), 5.36 (bs, 1H, 11-OH), 3.96 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.82 (bs, 3H, H-17), 0.59 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 659 [M.sup.+ + H] 32 12-epi-12-desvinyl-14-O-{[4-(3-Amino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(3- amino-propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.85 (d, 2H, arom., J = 8 Hz), 7.39 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H- 14, H = 6 Hz), 1.36 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6 Hz), 0.59 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 665 [M.sup. + Cl] 33 12-epi-12-desvinyl-14-O-{(4-[(3- Aminopropylcarbamoyl)-phenylsulfanyl)-acetyl}- 12-[(4-aminomethyl-benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.84 (d, 2H, arom., J = 8 Hz), 7.68 (d, 2H, arom., J = 6 Hz), 7.55 (d, 2 H, arom., J = 10 Hz), 7.37 (d, 2H, arom., J = 8 Hz), 5.43 (d, 1H, H-14, J = 7 Hz), 5.31 (bs, 1H, 11-OH), 1.34 (s, 3H, H-15), 0.83-0.77 (m, 6H, H-17, H-18), 0.57 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 693 [M.sup.+ + H] 34 12-epi-12-desvinyl-14-O-{(4-[(3- Aminopropylcarbamoyl)-phenylsulfanyl)-acetyl}- 12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.84 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H- 14, J = 6 Hz), 5.35 (bs, 1H, 11-OH), 1.35 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.4 Hz), 0.58 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 673 [M.sup.+ + H] 00 35 12-epi-12-desvinyl-14-O-{[4-(3- Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}- 12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.81 (t, 1H, CONH, J = 5.4 Hz), 7.84 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H-14, J = 6.8 Hz), 3.96 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 715 [M.sup.+ + H] 01 02 36 12-epi-12-desvinyl-14-O-{[4-(3- Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}- 12-[(8-amino-octylamino)-methyl]mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.84 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H- 14, J = 6.6 Hz), 3.96 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.79 (d, 3H, H-17, J = 5 Hz), 0.58 (d, 3H, H-16, J = 5 Hz). MS m/e: 701 [M.sup.+ + H] The reductive amination was done as described in Example 1, step 4 using 10 eq of 1,8-diaminooctane 03 04 37 12-epi-12-desvinyl-14-O-{[4-(3- Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}- 12-[(10-amino-decylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.84 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H- 14, J = 7 Hz), 3.96 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.79 (d, 3H, H-17, J = 5 Hz), 0.58 (d, 3H, H-16, J = 5 Hz). MS m/e: 729 [M.sup.+ + H] The reductive amination was done as described in Example 1, step 4 using 10 eq of 1,10-diaminodecane 05 06 38 12-epi-12-desvinyl-14-O-{(4-Carbamoyl- phenylsulfanyl)-acetyl}-12-[(6-guanidino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.82 (d, 2H, arom., J = 8 Hz), 7.37 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H- 14, J = 7 Hz), 3.96 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 658 [M.sup.+ + H] The required 4-mercapto-benzamide is e.g. described in: WO 2003/062235 07 08 39 12-epi-12-desvinyl-14-O-{[4-(3-Amino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12- {[3-(3-amino-propoxy)-propylamino)]-methyl} mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.83 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.44 (bs, 2H, H-14, 11-OH), 3.96 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6 Hz), 0.58 (d, 3H, H-16, J = 5.4 Hz); MS m/e: 689 [M.sup.+ + H] 09 0 40 12-epi-12-desvinyl-14-O-{[4-(3-Amino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12[(2- pyridin-4-yl-ethylamino)-methyl] mutilin trihydrochloride MS m/e: 679 [M.sup.+ + H] 41 12-epi-12-desvinyl-14-O-{[4-(6-Amino- hexylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6- guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.76 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H- 14, J = 7.6 Hz), 3.95 (s, 2H, H-22), 1.34 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.8 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 757 [M.sup.+ + H] 42 12-epi-12-desvinyl-14-O-{[4-(2-Amino- ethylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6- guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.79 (d, 2H, arom., J = 8 Hz), 7.40 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H- 14, J = 7.2 Hz), 3.96 (s, 2H, H-22), 1.34 (s, 3H, H-15), 0.94 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6.4 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 701 [M.sup.+ + H] 43 12-epi-12-desvinyl-14-O-{[4-(3- Aminopropylcarbamoyl)-phenylsulfanyl]-acetyl}- 12-{[3-(4-aminomethyl-phenyl)-propylamino]- methyl} mutilin trihydrochloride MS m/e: 721 [M.sup.+ + H] The required [4-(3-Amino-propyl)-benzyl]-carbamic acid tert-butyl ester is described in: Bioorganic & Medicinal Chemistry Letters, 16(11), 2986-2990; 2006

(43) According, e.g. analogously, to the method as set out under Example 100 below, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 4 are prepared. Chemical characterisation data are also set out in Table 4.

(44) TABLE-US-00004 TABLE 4 Example R.sub.2EX R.sub.1EX 44 12-epi-12-desvinyl-14-O-{[(4-Aminomethyl- cyclohexyl)-methylsulfanyl]-acetyl}-12-[(6-amino- hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.46 (d, 1H, H- 14, J = 6.6 Hz), 5.39 (bs, 1H, 11-OH), 1.39 (s, 3H, H-15), 0.96 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.8 Hz), 0.64 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 622 [M.sup.+ + H] 0 45 12-epi-14-O-[(1-Methyl-piperidin-4-ylsulfanyl)- acetyl}-12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride MS m/e: 638 [M + NH.sub.4.sup.+] The required 1-Methyl-piperidine-4-thiol is described in: J.Med.Chem., 36(22), 3251-64; 1993 46 12-epi-14-O-[(Piperidin-4-ylsulfanyl)-acetyl]-12- [(6-amino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H- 14, J = 6.2 Hz), 5.35 (bs, 1H, 11-OH), 1.41 (s, 3H, H-15), 0.99 (s, 3H, H-18), 0.83 (d, 3H, H-17, J = 6 Hz), 0.65 (d, 3H, H-16, J = 5 Hz) MS m/e: 580 [M.sup.+ + H]

EXAMPLE 47

12-epi-12-desvinyl-14-O-{[(4-Aminomethyl-cyclohexyl)-sulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(45) ##STR00125##
was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(46) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.47 (d, 1H, H-14, J=7.8 Hz), 1.40 (s, 3H, H-15), 0.98 (s, 3H, H-18), 0.82 (d, 3H, H-17, J=5.8 Hz), 0.64 (d, 3H, H-16, J=5 Hz)

(47) MS m/e: 608 [M.sup.++H]

Preparation of the required Thioacetic acid S-[4-(tert-butoxycarbonylamino-methyl)-cyclohexyl] ester

Step a: Methanesulfonic acid 4-(tert-butoxycarbonylamino-methyl)-cyclohexyl ester

(48) To a solution of 71 g of (4-Hydroxy-cyclohexylmethyl)-carbamic acid tert-butyl ester and 93.9 g of TEA in 1500 mL of DCM was added dropwise 34.1 g of MsCl at 0 C. over 30 min. After addition, the mixture was allowed to warm to rt and stirred at this temperature for 2.5 h. The reaction mixture was quenched by addition of water, the resulting mixture was extracted 3 with DCM. The combined organic layers were washed 3 with 0.5 M citric acid, 2 with a saturated solution of NaHCO.sub.3, water and dried over Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure to afford the title compound as yellow oil, and the crude product was used directly for the next step.

Step b: Thioacetic acid S-[4-(tert-butoxycarbonylamino-methyl)-cyclohexyl]ester

(49) A suspension of 52 g of methanesulfonic acid 4-(tert-butoxycarbonylamino-methyl)-cyclohexyl ester and 48.3 g of potassium thioacetate in 1 L of dry DMF was heated to 50 C. under N.sub.2 and stirred for 16 h. TLC showed the reaction was completed. The reaction mixture was cooled to 25 C. and poured into ice-water, the resulting mixture was extracted 5 with MTBE. The combined organic layers were washed 3 with citric acid, 2 with saturated solution of NaHCO.sub.3, 2 with brine and dried over Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduce pressure to afford the crude product as red oil. The crude product was purified by silica gel chromatography (PE/EtOAc=40:1 to 15:1) to afford the title compound as red solid.

EXAMPLE 48

12-epi-12-desvinyl-14-O-{[4-(3-Amino-propyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(50) ##STR00126##

Step 1: 12-epi-12-desvinyl-14-O-{{4-[3-(2,2,2-Trifluoro-acetylamino)-propyl]-phenylsulfanyl}-acetyl}-12-[(6-tert-butoxycarbonylamino-hexylamino)-methyl]mutilin

(51) The compound was prepared in analogy to compound 1, step 5 using 2,2,2-Trifluoro-N-[3-(4-mercapto-phenyl)-propyl]-acetamide as intermediate.

Step 2: 12-epi-12-desvinyl-14-O-{{4-[3-(2,2,2-Trifluoro-acetylamino)-propyl]-phenylsulfanyl}-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin dihydrochloride

(52) ##STR00127##

(53) The compound was prepared in analogy to compound 1, step 6.

Step 3: 12-epi-12-desvinyl-14-O-{[4-(3-Amino-propyl)-phenylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(54) ##STR00128##

(55) 246 mg of 12-epi-12-desvinyl-14-O-{{4-[3-(2,2,2-Trifluoro-acetylamino)-propyl]-phenylsulfanyl}-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin was dissolved in 10 mL of MeOH/H.sub.2O under ice-cooling and 0.51 mL of 1N NaOH was added. The reaction was stirred for 1 h under ice-cooling, poured into water and extracted with 3 with EtOAc. The organic phase was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The resulting colorless foam was purified by reversed phase chromatography (eluant: acetonitrile in 1N HCl 0-35%) yielding the title compound.

(56) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.58 (d, 2H, arom., J=5.6 Hz), 7.49 (d, 2H, arom., J=14 Hz), 4.43 (d, 1H, H-14, J=6.6 Hz), 1.34 (s, 3H, H-15), 0.84 (s, 3H, H-18), 0.79 (d, 3H, H-17, J=5.4 Hz), 0.54 (d, 3H, H-16, J=5.6 Hz)

(57) MS m/e: 630 [M.sup.++H]

Preparation of the required 2,2,2-Trifluoro-N-[3-(4-mercapto-phenyl)-propyl]-acetamido

Step a: 2,2,2-Trifluoro-N-(3-phenyl-propyl)-acetamide

(58) To a solution of 50 g of benzenepropanamine in 600 mL of MeOH was added dropwise 300 mL of CF.sub.3COOEt over a period of 0.5 h at 0 C. After stirring at 20 C. for 16 h, the solvent was removed under reduced pressure. The crude product was washed with PE to give the title compound as white solid.

Step b: 4-[3-(2,2,2-Trifluoro-acetylamino)-propyl]-benzenesulfonyl chloride

(59) To a solution of 32 g of 2,2,2-Trifluoro-N-(3-phenyl-propyl)-acetamide in 32 mL of DCM was added dropwise 200 mL of ClSO.sub.3H below 10 C. Then the reaction mixture was warmed to 15 C. and stirred for 20 h. The reaction mixture was poured into ice water at 05 C. and extracted 2 with DCM. The organic phase was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the crude product which was purified by silica gel chromatography (PE/EtOAc=10:1) to afford the title compound as yellow solid.

Step c: 2,2,2-Trifluoro-N-[3-(4-mercapto-phenyl)-propyl]-acetamide

(60) To a solution of 100 mL of H.sub.2SO.sub.4 in 600 mL of water was added slowly at 0 C. 102 g of Zinc dust and a solution of 49 g of 4-[3-(2,2,2-Trifluoro-acetylamino)-propyl]-benzenesulfonyl chloride in 50 mL of THF. Then the reaction mixture was heated to 60 C. and stirred for 4 h after which it was cooled and poured into DCM. The mixture was filtered and separated. The organic phase was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the crude product which was washed with PE/EtOAc=40:1 to afford the title compound as white solid.

EXAMPLE 49

(61) 12-epi-12-desvinyl-14-O-{(4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-{[(3-amino-propyl)-acetylamino]-methyl} mutilin dihydrochloride was prepared analogously to the method as set out under Example 1, step 6, but using the appropriate starting materials.

(62) ##STR00129##

(63) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.29-7.22 (m, 4H, arom.), 5.44 (d, 1H, H-14, J=5.6 Hz), 3.78 (s, 2H H-22), 2.05 (s, 3H, N-acetyl), 1.30 (s, 3H, H-15)

(64) MS m/e: 687 [M.sup.++H]

Preparation of the intermediate 12-epi-12-desvinyl-14-O-{([4-(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-{[(3-tert-butoxycarbonylamino-propyl)-acetylamino]-methyl} mutilin

(65) 500 mg of 12-epi-12-desvinyl-14-O-{([4-(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin (Example 1, step 5) was dissolved in 7 mL of DCM. To the mixture obtained 134 l of diisopropylethylamine was added and the mixture obtained was cooled in an ice bath, treated with 61.5 l of f acetic anhydride in 1 mL of DCM and stirring at rt was maintained for 1 h. To the mixture obtained 12 l of base and 6 l of acetic anhydride was added and stirring was continued for 30 min and the reaction was deemed to be complete. The mixture obtained was washed with 5% NaHCO.sub.3, the aqueous phase was back-extracted with DCM, the combined organic phases obtained were washed with water and brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness to yield the title compound.

EXAMPLE 50

12-epi-12-desvinyl-14-O-{(4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl)-acetyl}-12-(3-amino-propylcarbamoyl) mutilin dihydrochloride

Step 1: Pleuromutilintosylate-12-epi-12-desvinyl-12-carboxylic acid

(66) ##STR00130##

(67) 2.5 g of 12-epi-2-desvinyl-14-O-[(toluene-4-sulfonyloxy)-acetyl]-12-formyl mutilin (Example 1, step 3) was dissolved in 50 mL of DMF, 14.4 g of oxone was added and the resulting slurry was stirred at rt for 4 d. The mixture obtained was diluted with 300 mL of water, 50 mL 2N HCl was added and the mixture obtained was extracted with EtOAc (2). The phases obtained were separated and the combined organic phases were washed with water, dried over Na.sub.2SO.sub.4, brought to dryness (semisolid residue) and subjected to column chromatography (eluent: EtOAc/toluene=2:1). The title compound was obtained in the form of colourless crystals.

(68) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.79 (d, 2H, arom., J=8 Hz), 7.44 (d, 2H, arom., J=8 Hz), 5.41 (d, 1H, H-14, J=7.2 Hz), 4.70 AB-system (2H, H-22, v.sub.A=4.78, v.sub.B=4.70, J=16 Hz), 2.4 (s, 3H, tosyl-CH.sub.3), 1.30 (s, 3H, H-15), 1.08 (s, 3H, H-18), 0.80 (d, 3H, H-17, J=5 Hz), 0.52 (d, 3H, H-16, J=5.6 Hz).

Step 2: 12-epi-12-desvinyl-14-O-[(Toluene-4-sulfonyloxy)-acetyl]-12-(3-tert-butoxycarbonylamino-propylcarbamoyl) mutilin

(69) ##STR00131##

(70) 500 mg of pleuromutilintosylate-12-epi-12-desvinyl-12-carboxylic acid, 158 mg of N-BOC-1,3-diaminopropane, 139 mg of HOBt and 174 mg of N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride was dissolved in 10 mL of DCM and the mixture obtained was stirred at rt for 2 h. From the mixture obtained the phases were separated and the organic phase obtained was washed successively with 10% aqueous K.sub.2CO3, (2), 1N HCl (1) and 5% aqueous NaHCO.sub.3 solution (1), dried over Na.sub.2SO.sub.4, evaporated to dryness and the evaporation residue obtained was subjected to chromatography over silica gel with toluene/EtOAc=2:1, 1:1, 1:2, 1:5 and EtOAc. The title compound was obtained in the form of a colourless foam.

(71) MS m/e: 729 [M+Na]

Step 3: 12-epi-14-O-{{[(3-tert-Butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-(3-tert-butoxycarbonylamino-propylcarbamoyl) motilin

(72) ##STR00132##

(73) 314 mg of S-{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenyl} thioacetic acid ester was dissolved in 10 mL of MeOH/H.sub.2O 4:1, 229 l of 5 M aqueous K.sub.2CO.sub.3 solution was added and after 5 minutes 404 mg of 12-epi-12-desvinyl-14-O-[(Toluene-4-sulfonyloxy)-acetyl]-12-(3-tert-butoxycarbonylamino-propylcarbamoyl) mutilin. The mixture obtained was stirred for 15 min, diluted with 75 mL of water and extracted with EtOAc (2). The combined organic layers obtained were washed with 2N NaOH (2), 1N HCl and 5% NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and evaporated to dryness. Pale yellow crystals were obtained which were subjected to chromatography (eluent: EtOAc/toluene=2:1). The title compound was obtained in the form of colourless crystals.

(74) MS m/e: 859 [M.sup.++H].

Step 4: 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-{[(3-amino-propylcarbamoyl) mutilin dihydrochloride

(75) ##STR00133##

(76) BOC-deprotection was carried out as described in Example 1, Step 6.

(77) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.38 (t, 1H, CONH, J=4 Hz), 7.25 (d, 2H, arom., J=8 Hz), 7.17 (d, 2H, arom., J=8 Hz), 5.39 (d, 1H, H-14, J=7.4 Hz), 1.31 (s, 3H, H-15), 1.04 (s, 3H, H-18), 0.79 (d, 3H, H-17, J=6 Hz), 0.57 (d, 3H, H-16, J=5.8 Hz);

(78) MS m/e: 659 [M.sup.++H].

(79) According, e.g. analogously, to the method as set out under Example 50 above, but using appropriate starting materials, the compound of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 5 is prepared. Chemical characterisation data are also set out in Table 5.

(80) TABLE-US-00005 TABLE 5 Example R.sub.2EX R.sub.1EX 51 12-epi-12-desvinyl-14-O-{[4-(3-Amino- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-(4- aminomethyl-benzylcarbamoyl) mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.74 (t, 1H, PhCONH, J = 6 Hz), 8.17 (bs, 1H, CONH), 7.79 (d, 2H, arom., J = 8 Hz), 7.41-7.35 (m, 4H, arom.), 7.26 (d, 2H, arom., J = 8 Hz), 5.44 (d, 1H, H-14, J = 6.8 Hz), 4.62 (d, 1H, 11-OH, J = 6.8 Hz), 4.40-4.04 (m, 2H, PhCH.sub.2), 1.31 (s, 3H, H-15), 1.12 (s, 3H, H- 18), 0.81 (d, 3H, H-17, J = 6 Hz), 0.59 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 707 [M.sup.+ + H]

EXAMPLE 52

12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[2-(3-amino-propylamino)-ethyl] mutilin trihydrochloride

Step 1: 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(81) ##STR00136##

(82) 59.1 g of 12-epi-Pleuromutilintosylate was dissolved/suspended in I L of MeOH, 66.8 mL of trimethylorthoformate was added and the mixture obtained was cooled in an ice-bath. 13.15 mL of H.sub.2SO.sub.4 conc. was added while keeping the temperature below 10 C. The mixture obtained was stirred for further 30 min at this temperature, the ice bath was removed and the mixture obtained was stirred for a further 120 h. 21.5 g of solid NaHCO.sub.3 was added followed by addition of water until there was no more gas evolution. A precipitate formed and was filtered off, washed with water and the mother liquors obtained were extracted with DCM. The precipitate and the extracts were combined and dried.

(83) The synthesis of 4-epi-3-methoxy-pleuromutilin is e.g. disclosed in: Tetrahedron, 36, 1807 (1980).

Step 2: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(84) ##STR00137##

(85) 1.5 g of 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin was dissolved in 15 mL of dry THF and 10.8 mL of 0.5 M 9-Borabicyclo[3.3.1]nonane was added at rt. The resulting mixture was refluxed for 5 min, cooled to rt over 30 min and, eventually in an ice bath; 600 l of 35% H.sub.2O.sub.2 and 600 l of 3N NaOH was added. The mixture obtained was allowed to warm to rt over 30 min and the resulting slurry was partitioned between water and EtOAc. The phases were separated, the organic phase obtained was washed with water, dried over Na.sub.2SO.sub.4, evaporated to dryness and the evaporation residue was subjected to chromatography (eluent: toluene/acetone=10:1). The title compound in the form of a colourless solid was obtained.

(86) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.83 (d, 2H, arom., J=8 Hz), 7.48 (d, 2H, arom., J=8 Hz), 4.76, AB-system (2H, H-22, v.sub.A=4.80, v.sub.B=4.71, J=16 Hz), 4.40-4.35 (m, OH), 2.41 (s, 3H, tosyl-CH.sub.3), 1.18 (s, 3H, H-15), 1.08 (s, 3H, H-18), 0.88 (d, 3H, H-17, J=6 Hz), 0.63 (d, 3H, H-16, J=6.2 Hz).

Step 3: 4-epi-12-epi-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo mutilin

(87) ##STR00138##

(88) 430 mg of S-{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenyl} thioacetic acid ester was dissolved in 10 mL of MeOH, 470 l of 5 M aqueous K.sub.2CO.sub.3 solution was added and stirring was maintained for 10 min. To the mixture obtained 663 mg 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin in 2 mL of MeOH was added and the resulting yellow mixture was stirred for 2 h. From the mixture obtained solvent was evaporated and the evaporation residue was partitioned between EtOAc and brine. The phases were separated, the organic phase obtained was dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound was obtained in the form of a yellow foam.

(89) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.06-8.02 (m, 1H, CONH), 7.29 (d, 2H, arom., J=8 Hz), 7.18 (d, 2H, arom., J=8 Hz), 5.63 (d, 1H, H-14, J=10 Hz), 4.39-4.35 (m, 1H, OH), 3.84 (bs, 2H, H-22), 1.10 (s, 3H, H-18), 0.87 (d, 3H, H-17, J=5.8 Hz), 0.67 (d, 3H, H-16, J=6.4 Hz).

Step 4: 4-epi-12-epi-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl) mutilin

(90) ##STR00139##

(91) 200 mg of 4-epi-12-epi-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo mutilin was dissolved in 10 mL of DCM with stirring, 118 mg Dess-Martin-periodinane was added and stirring was maintained overnight. From the mixture obtained the phases were separated, the organic phase was washed with 5% aqueous NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and brought to dryness. The title compound in the form of colourless foamy crystals was obtained which was used without purification for the next step.

Step 5: 4-epi-12-epi-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[2-(3-tert-butoxycarbonylamino-propylamino)-ethyl]-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin

(92) ##STR00140##

(93) 200 mg of 4-epi-12-epi-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl) mutilin and 50 mg of N-BOC-1,3-diaminopropane were dissolved in 5 mL of DCM, 0.5 g of molecular sieves was added and stirring was maintained for 3 h. To the mixture obtained 150 mg of sodium triacetoxyborohydride was added and stirring was continued overnight. From the mixture obtained the phases were separated, the organic phase obtained was diluted with DCM, washed with 5% aqueous NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound was obtained in the form of colourless crystals which were used without purification for the next step. MS m/e: 874 [M.sup.++H].

Step 6: 12-epi-12-desvinyl-14-O-{{4-[(3-amino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-12-[2-(3-amino-propylamino)-ethyl] mutilin trihydrochloride

(94) ##STR00141##

(95) BOC-deprotection of the product obtained in step 5, which resulted also in methyl ether cleavage and retro hydride shift as described in Tetrahedron, 36, 1807 (1980), was carried out as described in Example 1, Step 6.

(96) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.28 (d, 2H, arom., J=8 Hz), 7.23 (d, 2H, arom., J=8 Hz), 5.44 (d, 1H, H-14, J=7 Hz), 4.71 (d, 1H, 11-OH, J=4.4 Hz), 1.33 (s, 3H, H-15), 0.78 (bs, 6H, H-17, H-18), 0.58 (d, 3H, H-16, J=5.8 Hz); MS m/e: 659 [M.sup.++H].

(97) According, e.g. analogously, to a method as set out under Example 1, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 6 are prepared. Chemical characterisation data are also set out in Table 6.

(98) TABLE-US-00006 TABLE 6 Example R.sub.2EX R.sub.1EX 53 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(3-amino- propylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.31-7.12 (m, 4 H, arom.), 5.45 (d, 1H, H-14, J = 6.6 Hz), 4.45 (s, 2H, PhCH.sub.2), 3.82 (s, 2H, H-22), 1.36 (s, 3H, H- 15), 0.88 (s, 3H, H-18), 0.81 (d, 2H, H-17, J = 5.6 Hz), 0.58 (d, 3H, H-16, J = 4.8 Hz) MS m/e: 561 [M.sup.+ + H] 54 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(6-amino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.31-7.11 (m, 4 H, arom.), 5.44 (d, 1H, H-14, J = 7.8 Hz), 4.45 (s, 2H, PhCH.sub.2), 3.82 (s, 2H, H-22), 1.36 (s, 3H, H- 15), 0.87 (s, 3H, H-18), 0.78 (d, 2H, H-17, J = 6.6 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 603 [M.sup.+ + H] 55 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.30-7.08 (m, 8H arom.), 5.47 (d, 1H, H-14, J = 7 Hz), 4.42 (s, 2H, PhCH.sub.2OH), 1.31 (s, 3H, H-15), 0.80 (s, 3H, H-18), 0.80-0.77 (6H, H-17, H-18), 0.59 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 623 [M.sup.+ + H] 56 12-epi-12-desvinyl-14-O-[(3-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(6-guanidino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.39-7.11 (m, 4 H, arom.), 5.44 (d, 1H, H-14, J = 7.6 Hz), 4.45 (s, 2H, PhCH.sub.2), 3.82 (s, 2H, H-22), 1.36 (s, 3H, H- 15), 0.87 (s, 3H, H-18), 0.80 (d, 2H, H-17, J = 6.4 Hz), 0.58 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 645 [M.sup.+ + H] 57 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(3-amino- propylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.32 (d, 2H, arom., J = 8.4 Hz), 7.25 (d, 2H, J = 8.4 Hz), 5.44 (d, 1H, H-14, J = 6 Hz), 4.45 (s, 2H, PhCH.sub.2), 3.8 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.87 (s, 3H, H-18) 0.58 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 561 [M.sup.+ + H] 0 58 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.66 (d, 2H, arom., J = 8 Hz), 7.53 (d, 2H, arom., J = 8 Hz), 7.30 (d, 2H, arom., J = 8 Hz), 7.23 (d, 2H, arom., J = 8.2 Hz), 5.41 (d, 1H, H-14, J = 6.8 Hz), 1.34 (s, 3H, H- 15), 0.56 (d, 3H, H-16, J = 5.8 Hz). MS m/e: 623 [M.sup.+ + H] 59 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl- phenylsulfanyl)-acetyl]-12-[(6-amino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.32 (d, 2H, arom., J = 8 Hz), 7.25 (d, 2H, arom., J = 8 Hz), 5.43 (d, 1H, H-14, J = 7.4 Hz), 4.44 (s, 2H, PhCH.sub.2), 3.87 (s, 2H H-22), 1.35 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.8 Hz), 0.57 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 603 [M.sup.+ + H] 60 12-epi-12-desvinyl-14-O-[(3,5-Bis- hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3- amino-propylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.45 (d, 1H, H- 14, J = 6.6 Hz), 4.44 (s, 4H, 2 PhCH.sub.2), 1.40 (s, 3H, H-15), 3.81 s (H-22), 1.37 (s, 3H, H-15), 0.90 (s, 3H, H-18), 0.81 (d, 2H, H-17, J = 5.4 Hz), 0.59 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 591 [M.sup.+ + H] The required intermediate (3-Hydroxymethyl-5- mercapto-phenyl)-methanol was prepared in analogy to the synthesis of (3-Hydroxymethyl-4- mercapto-phenyl)-methanol as described in J. Antibiot. 51 (8), 722; (1998) 61 12-epi-12-desvinyl-14-O-{[(2,3,5,6-Tetrafluoro- 4-hydroxymethyl)-phenylsulfanyl]-acetyl}-12- [(4-aminomethyl-benzylamino)-methyl]- mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.67 (d, 2H arom., H = 8 Hz), 7.54 (d, 2H arom., J = 8 Hz), 5.40 (d, 1H, H-14, J = 6.4 Hz), 4.56 (s, 2H, PhCH.sub.2OH), 1.33 (s, 3H, H-15), 0.86 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 5.6 Hz), 0.53 (d, 3H, H-16, J = 6.2 Hz). MS m/e: 695 [M.sup.+ + H] The required intermediate (2,3,5,6-Tetrafluoro-4- mercapto-phenyl)-methanol was prepared by LAH-reduction of 2,3,5,6-Tetrafluoro-4- mercaptobenzoic acid 62 12-epi-12-desvinyl-14-O-{[(1R,2R,4R)-4-Amino- 2-hydroxy-cyclohexylsulfanyl]-acetyl}-12-[(3- amino-propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.31-7.12 (m, 4 H, arom.), 5.48 (d, 1H, H-14, J = 7.8 Hz), 1.40 (s, 3H, H-15), 1.00 (s, 3H, H-18), 0.83 (d, 2H, H-17, J = 6.2 Hz), 0.64 (d, 3H, H-16, J = 4.8 Hz). MS m/e: 568 [M.sup.+ + H] The required intermediate [3-((1R,3R,4R)-3- Hydroxy-4-mercapto-cyclohexylamino)-propyl]- carbamic acid tert-butyl ester is described in EP2399904 0 63 12-epi-12-desvinyl-14-O-{[(1R,2R,4R)-4-Amino- 2-hydroxy-cyclohexylsulfanyl]-acetyl}-12-[(4- aminomethyl-benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.67 (d, 2H, arom., J = 8 Hz), 7.53 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H-14, H = 7.4 Hz), 1.39 (s, 3H, H-15), 0.93 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6 Hz), 0.63 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 630 [M.sup.+ + H] 64 12-epi-12-desvinyl-14-O-[(2-Hydroxy- ethylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.67 (d, 2H arom., J = 8 Hz), 7.54 (d, 2H arom., J = 8 Hz), 5.46 (d, 1H, H-14, J = 7 Hz), 1.40 (s, 3H, H-15), 0.93 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 6.2 Hz), 0.65 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 561 [M.sup.+ + H] 65 12-epi-12-desvinyl-14-O-[(2-Amino- ethylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.69 (d, 2H arom., J = 8 Hz), 7.54 (d, 2H arom., J = 8 Hz), 5.47 (d, 1H, H-14, J = 6.8 Hz), 1.41 (s, 3H, H-15), 0.95 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.8 Hz), 0.65 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 560 [M.sup.+ + H] 66 12-epi-12-desvinyl-14-O-{[(5-Amino-4H-1,2,4- triazol-3-yl)-sulfanyl]-acetyl}-12-[(6-amino- hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.47 (d, 1H, H- 14, J = 7 Hz), 5.33 (bs, 1H, 11-OH), 3.93 (s, 2H, H- 22), 1.38 (s, 3H, H-15), 0.90 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6.2 Hz), 0.60 (d, 3H, H-16, J = 5.2 Hz). MS m/e: 579 [M.sup.+ + H] 67 12-epi-12-desvinyl-14-O-[(2-Amino- ethylsulfanyl)-acetyl]-12-[(6-amino- hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.49 (d, 1H, H- 14, J = 6.6 Hz), 3.77 (s, 2H, H-22), 1.42 (s, 3H, H- 15), 1.01 (s, 3H, H-18), 0.83 (d, 3H, H-17, J = 6.2 Hz), 0.66 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 540 [M.sup.+ + H] 0 68 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)- acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.36 (d, 2H, arom., J = 5.4 Hz), 7.27 (d, 2H, arom., J = 5.2 Hz), 5.52 (d, 1H, H-14, J = 6.6 Hz), 4.02 (s, 2H, H-22), 1.31 (s, 3H, H-15), 0.85 (s, 3H, H-18), 0.77 (d, 3H, H-17, J = 6.2 Hz), 0.60 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 574 [M.sup.+ + H] 69 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)- acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.39 (d, 2H, arom., J = 3.8 Hz), 7.29 (d, 2H, arom., J = 5 Hz), 5.50 (d, 1H, H-14, J = 6.8 Hz), 4.06 (s, 2H, H-22), 1.37 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6.4 Hz), 0.61 (d, 3H, H-16, J = 5.4 Hz). MS m/e: 616 [M.sup.+ + H] 70 12-epi-12-desvinyl-14-O-[(3-Hydroxy- phenylsulfanyl)-acetyl]-12-[(6-amino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.07 (m, 1H, arom.), 7.71 (m, 2H, arom.), 6.57 (m, 1H, arom.), 5.48 (d, 1H, H-14, J = 7 Hz), 3.73 (s, 2H, H-22), 1.30 (s, 3H, H-15), 0.83 (s, 3H, H-18), 0.77 (d, 3H, H-17, J = 6.6 Hz), 0.58 (d, 3H, H-16, J = 5.2 Hz). MS m/e: 589 [M.sup.+ + H] 71 12-epi-12-desvinyl-14-O-[(4-Fluoro- phenylsulfanyl)-acetyl]-12-[(6-amino- hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.44 (m, 2H, arom.), 7.17 (m, 2H, arom.), 5.46 (d, 1H, H-14, J = 7.8 Hz), 3.79 (m, 2H, H-22), 1.28 (s, 3H, H-15), 0.83 (s, 3H, H-18), 0.78 (d, 3H, H-17, J = 6.6 Hz), 0.56 (d, 3H, H-16, J = 5.6 Hz). MS m/e: 591 [M.sup.+ + H] 72 12-epi-12-desvinyl-14-O-{[(7H-Purin-6-yl)- sulfanyl]-acetyl}-12-[(6-amino-hexylamino)- methyl] mutilin dihydrochloride MS m/e: 615 [M.sup.+ + H] 0 73 12-epi-12-desvinyl-14-O-[(3-Amino- phenylsulfanyl)-acetyl]-12-[(3-amino- propylamino)-methyl] mutilin trihydrochloride MS m/e: 546 [M.sup.+ + H] 74 12-epi-12-desvinyl-14-O-(Phenylsulfanyl- acetyl)-12-[(4-aminomethyl-benzylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.68 (d, 2H, arom., J = 8 Hz), 7.56 (d, 2H, arom., J = 8 Hz), 7.33-7.23 (m, 5H, arom.), 5.42 (d, 1H, H-14, J = 6.4 Hz), 3.83 (bs, 2H, H-22), 1.34 (s, 3H, H- 15), 0.79 (bs, 6H, H-18, H-17), 0.55 (d, 3H, H- 16, J = 5.8 Hz) MS m/e: 593 [M.sup.+ + H] 75 12-epi-12-desvinyl-14-O-[(4-Fluoro- phenylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin trihydrochloride MS m/e: 611 [M.sup.+ + H] 76 12-epi-12-desvinyl-14-O-[(Pyridin-2-ylsulfanyl)- acetyl]-12-[(4-aminomethyl-benzylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.32-8.30 (m, 1H, arom., J = 4.4 Hz), 7.70-7.54 (m, 5H, arom.), 7.36-7.32 (m, 1H, arom.), 7.12-7.06 (m, 1H, arom.), 5.45 (d, 1H, H-14, J = 7.8 Hz), 5.31 (bs, 1H, 11-OH), 3.94 (bs, 2H, H-22), 1.37 (s, 3H, H-15), 0.84-0.78 (m, 6H, H-18, H-17), 0.62 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 594 [M.sup.+ + H] 77 12-epi-12-desvinyl-14-O-[(Pyridin-4-ylsulfanyl)- acetyl]-12-[(4-aminomethyl-benzylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.56 (m, 2H, arom.), 7.67 (d, 2H, arom., J = 8 Hz), 7.54 (d, 2H, arom., J = 8 Hz), 7.48-7.45 (m, 2H, arom.), 5.46 (d, 1H, H-14, J = 7.2 Hz), 5.28 (d, 1H 11- OH, J = 4.8 Hz), 4.14 (bs, 2H, H-22), 1.34 (s, 3H, H-15), 0.85 (s, 3H, H-18), 0.78 (d, 3H, H- 17, J = 6.4 Hz), 0.58 (d, 3H, H-16, J = 6 Hz) MS m/e: 594 [M.sup.+ + H] 0

EXAMPLE 78

(99) 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(100) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.68 (d, 2H, arom., J=8 Hz), 7.54 (d, 2H, J=8 Hz), 5.46 (d, 1H, H-14, J=7.2 Hz), 5.32 (d, 1H, 11-OH, J=4 Hz), 1.37 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.81, (d, 3H, H-17, J=6.2 Hz), 0.64 (d, 3H, H-16, J=6 Hz)

(101) MS m/e: 657 [M.sup.++H]

(102) ##STR00192##

Preparation of the required Thioacetic acid S-[1-(2-tert-butoxycarbonylamino-acetyl)-piperidin-4-yl] ester Step a: 4-Methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester

(103) To a mixture of 10 g of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester nd 12.57 g of TEA in 100 mL of DCM was added 7.09 g of MsCl dropwise at 0 C. under N.sub.2. After addition, the resulting mixture was stirred at 20 C. for 4 hours. TLC showed the reaction was complete. The mixture was washed 2 with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound as white solid.

Step b: 4-Acetylsulfanyl-piperidine-1-carboxylic acid tert-butyl ester

(104) To a solution of 13.50 g of 4-methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester in 140 mL of DMF was added 11.04 g of potassium ethanethioate in one portion at 20 C. under N.sub.2. The mixture was stirred at 20 C. for 10 min. Then the reaction mixture heated to 60 C. and stirred for 16 hours. TLC showed the reaction was complete. The mixture was cooled to 20 C. and poured into ice-water and stirred for 20 min. The aqueous phase was extracted 2 with MTBE. The combined organic phase was washed 2 with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=30/1, 20/1) to afford the title compound as red oil.

Step c: Thioacetic acid S-piperidin-4-yl ester hydrochloride

(105) A solution of 6.5 g of 4-acetylsulfanyl-piperidine-1-carboxylic acid tert-butyl ester in 100 mL of EtOAc was acidified by adding a solution of HCl/EtOAc dropwise at 20 C. over a period of 30 min. under N.sub.2. The resulting mixture was stirred at 20 C. for 0.5 h after which a solid formed. TLC (PE:EtOAc=10:1) showed the reaction was complete. The reaction mixture was filtered to afford the title compound as off-white solid.

Step d: Thioacetic acid S-[1-(2-tert-butoxycarbonylamino-acetyl)-piperidin-4-yl]ester

(106) To a mixture of 4.70 g of thioacetic acid S-piperidin-4-yl ester hydrochloride, 3.84 g of N-BOC-Glycine and 7.29 g of DIPEA in 50 mL DMF was added 10.96 g of HATU in one portion at 20 C. under N.sub.2. The mixture was stirred at 20 C. for 16 h. The residue was poured into ice-water and stirred for 20 min. The aqueous phase was extracted 3 with MTBE. The combined organic phase was washed 2 with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by chromatography (PE/EtOAc=30/1, 20/1) to give the title compound as yellow oil.

(107) According, e.g. analogously, to a method as set out under Example 78 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 7 are prepared. Chemical characterisation data are also set out in Table 7.

(108) TABLE-US-00007 TABLE 7 Example R.sub.2EX R.sub.1EX 79 12-epi-12-desvinyl-14-O-{[1-(3-Amino-propionyl)- piperidin-4-yl-sulfanyl)]-acetyl}-12-[(4- aminomethyl-benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.67 (d, 2H, arom., J = 8 Hz), 7.53 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H-14, J = 7 Hz), 5.31 (d, 1H 11-OH, J = 4.6 Hz), 1.40 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5 Hz), 0.64 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 611 [M.sup.+ + H] The required Thioacetic acid S-[1-(3-tert- butoxycarbonylamino-propionyl)-piperidin-4-yl] ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials 80 12-epi-12-desvinyl-14-O-{[1-(3-Amino-propionyl)- piperidin-4-yl-sulfanyl]-acetyl}-12-[(6-amino- hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H- 14, J = 7 Hz), 5.37 (bs, 11-OH), 1.41 (s, 3H, H- 15), 0.98 (s, 3H, H-18), 0.83 (d, 3H, H-17, J = 6 Hz), 0.65 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 325 [M/2.sup.+ + H] The required Thioacetic acid S-[1-(3-tert- butoxycarbonylamino-propionyl)-piperidin-4-yl] ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials 81 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)- piperidin-4-yl-methylsulfanyl]-acetyl}-12-[(4- aminomethyl-phenylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.51 (d, 1H, H- 14, J = 5.8 Hz), 1.39 (s, 3H, H-15), 0.99 (s, 3H, H- 18), 0.85 (d, 3H, H-17, J = 6.4 Hz), 0.65 (d, 3H, H- 16, J = 6.4 Hz) MS m/e: 691 [M.sup. + Cl] The required Thioacetic acid S-[1-(2-tert- butoxycarbonylamino-acetyl)-piperidin-4-ylmethyl] ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials 82 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)- piperidin-4-yl-methylsulfanyl]-acetyl}-12-[(4- amino-benzylamino)-methyl] mutilin trihydrochloride MS m/e: 656 [M.sup.+ + H] For the reductive amination N-(4-Aminomethyl- phenyl)-2,2,2-trifluoro-acetamide (obtained in 2 steps from (4-Amino-benzyl)-carbamic acid tert- butyl ester) was used. In the subsequent nucleophilic displacement reaction (e.g. Example 1, step 5) using K.sub.2CO.sub.3 instead of potassium tert-butoxide as base the N-trifluoroacetyl group was concomitantly cleaved 00

EXAMPLE 83

(109) 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12{2-[4-(2-amino-ethoxy)-benzylamino]-methyl} mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 1 and 78, but using appropriate starting materials.

(110) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.57 (d, 2H, arom., J=8 Hz), 7.00 (d, 2H, arom., J=8 Hz), 5.45 (d, 1H, H-14, J=6.2 Hz), 1.39 (s, 3H, H-15), 0.93 (s, 3H, H-18), 0.81 (d, 3H, H-17, J=6 Hz), 0.64 (d, 3H, H-16, J=5.4 Hz)

(111) MS m/e: 687 [M.sup.++H]

Preparation of the required [2-(4-Aminomethyl-phenoxy)-ethyl]-carbamic acid tert-butyl ester

(112) ##STR00201##

Step a: (4-Hydroxy-benzyl)-carbamic acid benzyl ester

(113) 10 g of 4-Aminomethyl-phenol was dissolved in 200 mL DMF, the solution was cooled to 0 C., 13.6 mL of triethylamine followed by 11.6 mL of benzyl chloroformate was added and stirring was continued at rt overnight. The reaction mixture was diluted with water and extracted 2 with EtOAc, the combined organic layers were washed 2 with 2N hydrochloric acid, 5% sodium bicarbonate solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness to give an almost colourless oil. The product was chromatographed over silica gel (eluant: toluene/EtOAc=10:1) and yielded the title compound as almost colourless oil.

Step b: {2-[4-(Benzyloxycarbonylamino-methyl)-phenoxy]-ethyl}-carbamic acid tert-butyl ester

(114) 3.25 g of (4-Hydroxy-benzyl)-carbamic acid benzyl ester, 3.11 g of (2-bromo-ethyl)-carbamic acid tert-butylester, 4.10 g of cesium carbonate and 209 mg of potassium iodide was refluxed in 20 mL 2-butanone for 24 hours. The solvent was removed in vacuo, the residue partitioned between water and EtOAc. The aqueous phase was washed with EtOAc, the combined organic layers with 2N sodium hydroxide solution and finally with brine. After drying over anhydrous Na.sub.2SO.sub.4 and evaporation of the solvent a yellowish oil was obtained which was chromatographed over silica gel (eluant: toluene/EtOAc=10:1) to yield an almost colorless viscous oil.

Step c: [2-(4-Aminomethyl-phenoxy)-ethyl]-carbamic acid tert-butyl ester

(115) 2.0 g of {2-[4-(Benzyloxycarbonylamino-methyl)-phenoxy]-ethyl}-carbamic acid tert-butyl ester, dissolved in 120 mL ethanol, was hydrogenated in an H-cube apparatus over 10% Pd/C at 10 bar and 50 C. for 3 days. The solvent was evaporated and the title compound (containing residual solvent) was obtained as a slowly crystallizing oil.

EXAMPLE 84

(116) 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[{4-[(2-amino-ethoxy)-benzylamino]-methyl}mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 1 and 83, but using appropriate starting materials.

(117) MS m/e: 715 [M.sup.++H]

(118) The required Thioacetic acid S-{4-[(2-tert-butoxycarbonylamino-acetylamino)-methyl]-cyclohexyl} ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials.

(119) ##STR00202##

EXAMPLE 85

12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride

Step 1: 12-epi-12-desvinyl-14-O-[(4-Azidomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(120) ##STR00203##

(121) 264 mg of 12-epi-12-desvinyl-14-O-[(4-hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin (Example 57, prior to BOC-deprotection) was dissolved in 4 mL of THF and the flask cooled in an ice bath. To the mixture obtained 122 mg of DPPA was added, followed by 76 mg of DBU in 3 mL of THF over five minutes. The cooling bath was removed, the mixture obtained was stirred at rt overnight and 45 mg of DPPA and 30.5 mg of DBU was further added.

(122) The mixture obtained was stirred for another 6 h. HPLC revealed still some starting material left; thus 56.8 mg of DPPA and 38.1 mg of DBU was again added, stirring was continued overnight and the reaction was deemed to be complete. About 40 mL of water was added and the mixture obtained was extracted with EtOAc (2), the combined organic phases were washed with 2N NaOH and water, dried over Na.sub.2SO.sub.4, evaporated to dryness and subjected to column chromatography over silica gel (eluent: EtOAc/Et.sub.3N=80:1). The title compound was obtained in the form of a foamy residue.

Step 2: 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(123) ##STR00204##

(124) 155 mg of 12-epi-12-desvinyl-14-O-[(4-azidomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin was dissolved in 4 mL of EtOH and 1.3 mL of water in an ultrasonic bath and 32.3 mg of NH.sub.4Cl and 20 mg of zinc powder was added. The mixture obtained was refluxed for 10 min. Since the reaction was not complete, 164 mg of NH.sub.4Cl and 100 mg of zinc powder was added and the mixture obtained was stirred at rt overnight. To the mixture obtained 75 mL of EtOAc and 5 mL of 25% ammonia solution was added, the formed precipitate was filtered off and the filtrate obtained was treated with brine, stirred for 5 min and from the mixture obtained the phases were separated. The organic phase obtained was dried over Na.sub.2SO.sub.4, evaporated to dryness and the evaporation residue was subjected to silica gel chromatography (eluent: EtOAc/EtOH/25% ammonia=100:2:0.5). The title compound was obtained in the form of colourless foamy crystals.

(125) Step 3: 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride BOC-deprotection of the product of step 2 above was carried out as described in Example 1, Step 6.

(126) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.45 (d, 2H, arom., J=8.2 Hz), 7.23 (d, 2H, arom., J=8.2 Hz), 5.46 (d, 1H, H-14, J=7 Hz), 3.88 (s, 2H, H-22), 1.37 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.81 (d, 3H, H-17, J=6.2 Hz), 0.59 (d, 3H, H-16, J=5.2 Hz)

(127) MS m/e: 594 [M.sup.+Cl]

(128) ##STR00205##

(129) According, e.g. analogously, to a method as set out under Example 85 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 8 are prepared. Chemical characterisation data are also set out in Table 8.

(130) TABLE-US-00008 TABLE 8 Example R.sub.2EX R.sub.1EX 86 12-epi-12-desvinyl-14-O-[(3-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(3-amino- propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.54 (s, 1H, arom.), 7.36-7.24 (m, 4H, 3H arom., 1H, NH), 5.46 (d, 1H, H-14, J = 6.4 Hz), 5.32 (bs, 1H, 11-OH), 3.93 (s, 2H, H-22), 1.35 (s, 3H, H-15), 0.90 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.8 Hz), 0.59 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 560 [M.sup.+ + H] 06 07 87 12-epi-12-desvinyl-14-O-[(3-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(6-guanidino- hexylamino)-methyl] mutilin trihydrochloride MS m/e: 645 [M.sup.+ + H] 08 09 88 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(4-aminomethyl- benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.68 (d, 2H arom., J = 8 Hz), 7.55 (d, 2H arom., J = 8 Hz), 7.49 (d, 2H arom., J = 8 Hz), 7.33 (d, 2 H arom., J = 8 Hz), 1.36 (s, 3H, H-15), 0.83 (s, 3H, H-18), 0.57 (d, 3H, H-16, J = 3.6 Hz) MS m/e: 622 [M.sup.+ + H] 0 89 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.46 (d, 2H arom., J = 8 Hz), 7.36 (d, 2H arom., J = 8 Hz), 5.48-5.39 (m, 2H, H-14, 11-OH), 1.38 (s, 3H, H-15), 0.90 (s, 3H, H- 18), 0.81 (d, 3H, H-17, J = 6.2 Hz), 0.60 (d, 3H, H-16, J = 5.4 Hz); MS m/e: 602 [M.sup.+ + H] 90 12-epi-12-desvinyl-14-O-[(3-Aminomethyl- phenylsulfanyl)-acetyl]-12-[((4-aminomethyl- cyclohexyl)-methylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.54 (s, 1H arom.), 7.34 (bs, 3H arom.), 5.48 (d, 1H, H-14, J = 7.2 Hz), 1.36 (s, 3H, H-15), 0.95 (s, 3H, H-18), 0.82 (d, 3H, H- 17, J = 6.2 Hz), 0.60 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 628 [M.sup.+ + H] 91 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-{[(4-aminocyclohexyl)- amino]-methyl} mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.30-7.16 (m, 4H arom.), 7.54 (d, 2H arom., J = 8 Hz), 5.48 (d, 1H, H-14, J = 7.4 Hz), AB (2H, H-22, v.sub.A = 3.85, v.sub.B = 3.76, J = 16 Hz), 1.29 (s, 3H, H-15), 0.83 (s, 3H, H-18), 0.77 (d, 3H, H-17, J = 6.4 Hz), 0.59 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 600 [M.sup.+ + H] 92 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(hexylamino)-methyl] mutilin dihydrochloride MS m/e: 587 [M.sup.+ + H] 93 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(4-carbamoylphenyl)- methylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.83 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 7.32-7.13 (m, 4H, arom.), 5.49 (bs, 1H, H-14), 1.31 (s, 3H, H-15), 0.79 (bs, 6H, H-17, H-18), 0.60 (bs, 3H, H-16) MS m/e: 636 [M.sup.+ + H] 0

EXAMPLE 94

(131) 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(3-amino-propylcarbamoyl)-benzylamino]-methyl} mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 85, but using appropriate starting materials.

(132) MS m/e: 727 [M.sup.++H]

(133) ##STR00222##

Preparation of the intermediate tert-butyl N-[3-[[4-(aminomethyl)-benzoyl]-amino]-propyl]-carbamate

Step a: [3-(4-Cyanobenzoylamino)-propyl]-carbamic acid tert-butyl ester

(134) A mixture of 1.47 g of 4-cyanobenzoic acid, 5.7 g of HATU and 2.58 g DIEA in 25 mL of DMF was stirred at 25 C. for 45 min. To the mixture obtained 3.48 g (3-Amino-propyl)-carbamic acid tert-butyl ester was added slowly and the mixture obtained was stirred at rt for another 2 h. The mixture obtained was quenched with water and extracted with EtOAc (30 mL*3). The phases obtained were separated and the organic phase was washed with water and brine, dried over Na.sub.2SO.sub.4, concentrated and the concentration residue was subjected to chromatography (DCM/MeOH=10:1) yielding the title compound.

Step b: [3-(4-Aminomethyl-benzoylamino)-propyl]-carbamic acid tert-butyl ester

(135) A mixture of 1 g of [3-(4-cyanobenzoylamino)-propyl]-carbamic acid tert-butyl ester and Pd/C (100 mg) in 25 mL of THF was stirred at 25 C. under hydrogen for 5 h. The mixture obtained was filtered and concentrated. The concentration residue was subjected to chromatography (DCM/MeOH=10:1) yielding the title compound.

EXAMPLE 95

(136) 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(5-dimethylcarbamoyl-pentylamino)-methyl] mutilin dihydrochloride was obtained similarly and in analogy to a method as set out in Example 85, but using appropriate starting materials.

(137) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.45 (d, 2H, arom., J=8 Hz), 7.34 (d, 2H, arom., J=8 Hz), 5.46 (d, 1H, H-14, J=6.4 Hz), 1.37 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.80 (d, 3H, H-17, J=6.4 Hz), 0.59 (d, 3H, H-16, J=5.6 Hz)

(138) MS m/e: 678 [M.sup.++H].

(139) ##STR00223##

Preparation of the intermediate 6-Amino-hexanoic acid dimethylamide

(140) Step a: A solution of 305 mg of Me.sub.2NH.HCl and 970 mg of DIPEA in 10 mL of DMF was stirred at rt for 30 min and 500 mg of N-benzyloxycarbonyl-6-aminocaproic acid and 1.07 g of HATU was added and the mixture obtained was stirred at rt overnight. The mixture obtained was extracted 3 with 10 mL EtOAc and the combined organic phases were washed with brine, dried and concentrated.

(141) Step b: 400 mg of the crude product of step a was dissolved in 15 mL of MeOH and Pd/C (wet, 100 mg) was added. The mixture obtained was stirred at rt under 30 Psi H.sub.2-pressure overnight. TLC showed that the reaction was completed. The mixture obtained was filtered and the filtrate was concentrated. The title compound in the form of a white solid was obtained and used without further purification.

EXAMPLE 96

(142) 12-epi-12-desvinyl-4-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(2-amino-2-carbamoyl-ethyl)-benzylamino]-methyl} mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 85, but using appropriate starting materials.

(143) MS m/e: 679 [M++H]

Preparation of the intermediate tert-butyl N-[2-Amino-1-[[4-(aminomethyl)-phenyl]-methyl]-2-oxo-ethyl]-carbamate

(144) ##STR00224##

Step a: 2-tert-Butoxycarbonylamino-3-(4-cyano-phenyl)-propionic acid methyl ester

(145) A mixture of 8.4 g of 4-cyano-phenylalanine methylester (prepared in analogy to JACS, 129(1), 14-15; 2007), 9 g of BOC.sub.2O and 50 mg of DMAP in 150 m, of THF was stirred at rt for 3.5 h, diluted with water, extracted with EtOAc and subjected to column chromatography (DCM/MeOH=100:4) yielding the title compound.

Step b: 2-tert-Butoxycarbonylamino-3-(4-cyano-phenyl)-propionic acid

(146) A mixture of 1.91 g of 2-tert-butoxycarbonylamino-3-(4-cyano-phenyl)-propionic acid methyl ester and 0.38 g NaOH in 30 mL of THF and 15 mL of water was stirred at rt for 2 h, concentrated to removed THF, extracted with EtOAc, acidified by HCl to pH 3, extracted with EtOAc and concentrated to yield the title compound.

Step c: [1-Carbamoyl-2-(4-cyano-phenyl)-ethyl]-carbamic acid tert-butyl ester

(147) A mixture of 0.9 g of 2-tert-Butoxycarbonylamino-3-(4-cyano-phenyl)-propionic acid, 1.413 g of HATU, and 1.6 g of DIPEA in 60 mL of DMF was stirred at rt for 2 h and 9.3 mmol of NH.sub.4Cl was added. The mixture was stirred at rt for 2 h, diluted with water, extracted with EtOAc and subjected to column chromatography (DCM/MeOH=100:5) yielding the title compound.

Step d: tert-butyl N-[2-Amino-1-[[4-(aminomethyl)-phenyl]-methyl]-2-oxo-ethyl]-carbamate

(148) A mixture of 1.58 mmol [1-carbamoyl-2-(4-cyano-phenyl)-ethyl]-carbamic acid tert-butyl ester, 50 mg of Ni and 50 mg of NH.sub.3H.sub.2O in 50 mL of MeOH was stirred at rt under H.sub.2 for 4 h, filtered and concentrated to yield the title compound.

EXAMPLE 97

(149) 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-{[4-(2-amino-2-dimethylcarbamoyl-ethyl)-benzylamino]-methyl} mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 85, but using appropriate starting materials.

(150) MS m/e: 707 [M.sup.++H]

Preparation of the intermediate tert-butyl N-[1-[[4-(Aminomethyl)-phenyl]-methyl]-2-(dimethylamino)-2-oxo-ethyl]-carbamate

(151) ##STR00225##

(152) Step a: A solution of 305 mg Me.sub.2NH.HCl and 970 mg in 10 mL of DMF was stirred at rt for 30 min. To the mixture obtained 545 mg of 2-tert-butoxycarbonylamino-3-(4-cyano-phenyl)-propionic acid (see example 96) and 1.07 g of HATU were added and the mixture obtained was stirred at rt overnight. The mixture obtained was extracted 3 with 10 mL EtOAc, the combined organic phases were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and used for the next step without further purification.

(153) Step b: A mixture 458 mg of the residue of step a, 50 mg of Ni and 50 mg of NH.sub.3H.sub.2O in 50 mL of MeOH was stirred at rt under H.sub.2 for 4 h, filtered and concentrated to yield the title compound.

EXAMPLE 98

(154) 12-epi-12-desvinyl-14-O-{[5-Aminomethyl-pyridin-2-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin tetrahydrochloride was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(155) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.83-7.78 (m, 1H, arom.), 7.70 (d, 2H, arom., J=8 Hz), 7.55 (d, 2H, arom., J=8 Hz), 7.41-7.37 (m, 1H, arom.), 5.45 (bs, 2H, H-14, 11-OH), 3.92 (s, 2H, H-22), 1.37 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.79 (s, 3H, H-17, J=6 Hz), 0.64 (d, 3H, H-16, J=5.6 Hz)

(156) MS m/e: 623 [M.sup.++H]

(157) ##STR00226##

Preparation of the required Thioacetic acid S-[5-(tert-butoxycarbonylamino-methyl)-pyridin-2-yl] ester

Step a: (6-Mercapto-pyridin-3-ylmethyl)-carbamic acid tert-butyl ester

(158) To a mixture of 12 g 6-mercapto-nicotinamide (described in PCT Int. Appl., 2011039259) in 250 mL THF was added 46.7 mL of BH.sub.3-Me.sub.2S (dropwise at 25 C. under N.sub.2. The mixture was stirred at 70 C. for 12 hr. The mixture was cooled to 0 C., 100 mL of 1N HCl) was added to the reaction and stirred at 45 C. for 1 h, the aqueous phase was extracted 3 with EtOAc, the aqueous phase was adjusted to pH=13 with 1N NaOH. To the solution was added 21.23 g of BOC.sub.2O and the mixture was stirred at 25 C. for 12 h. The aqueous phase was extracted 3 with EtOAc. The combined organic phase was washed 2 with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10:1 to 1:3) to afford the title compound as yellow solid.

Step b: Thioacetic acid S-[5-(tert-butoxycarbonylamino-methyl)-pyridin-2-yl]ester

(159) A mixture of 2.5 g of (6-mercapto-pyridin-3-ylmethyl)-carbamic acid tert-butyl ester in 25 mL DCM, was added 4.21 g of TEA at 0 C. under N.sub.2. The reaction was stirred at 0 C. for 10 min and 1.63 g of acetyl chloride was added drop-wise at 0 C. under N.sub.2. The mixture was stirred at 0 C. for 20 min. The mixture was poured into ice-water and stirred for 10 min. The aqueous phase was extracted 3 with DCM. The combined organic phase was washed with 2 brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE/EtOAe1:1) to afford the title compound as yellow solid.

EXAMPLE 99

(160) 12-epi-12-Desvinyl-14-O-{[5-aminomethyl-pyridin-2-yl-sulfanyl)]-acetyl}-12-[(4-aminomethyl-3-fluoro-benzylamino)-methyl] mutilin tetrahydrochloride was obtained similarly and in analogy to a method as set out in Example 98, but using appropriate starting materials.

(161) MS m/e: 641 [M.sup.++H]

Preparation of the required (4-Aminomethyl-2-fluoro-benzyl)-carbamic acid tert-butyl ester

(162) ##STR00227##

Step a: (4-Cyano-2-fluoro-benzyl)-carbamic acid tert-butyl ester

(163) To a mixture of 16 g of tert-butyl N-[(4-bromo-2-fluoro-phenyl)methyl]carbamate in 480 mL of DMF was added 12.35 g of Zn(CN), 5.83 g of dppf, 687.98 mg of Zn and 4.82 g of Pd.sub.2(dba).sub.3 under N.sub.2. The reaction mixture was heated to 120-128 C. and stirred for 5 hours. TLC showed the reaction was complete. The mixture was cooled to 25 C. and poured into ice-water and stirred for 20 min. The aqueous phase was extracted 3 with EtOAc. The combined organic phase was washed 2 with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (silica gel, PE/EtOAc=30/1 to 5/1) to afford the product as yellow solid.

Step b: (4-Aminomethyl-2-fluoro-benzyl)-carbamic acid tert-butyl ester

(164) To a solution of 9.00 g tert-butyl N-[(4-cyano-2-fluoro-phenyl)methyl]carbamate in 150 mL of MeOHNH.sub.3 was added 1.80 g of Raney Nickel under N.sub.2. The suspension was degassed in vacuum and purged with H.sub.2 several times. The mixture was stirred at 25 C. under H.sub.2 (50 psi) for 8 hours. TLC (PE/EtOAc=5:1) showed the starting material was consumed completely. The reaction mixture was filtered and the filtrate was concentrated in vacuum. The residue was purified by silica gel chromatography eluted with DCM/MeOH=100:1 to 10:1 to give the title compound as brown solid.

EXAMPLE 100

(165) 12-epi-12-desvinyl-14-O-{[(4-Aminomethyl-cyclohexyl)-methyl sulfanyl)-acetyl]{[(4-Aminomethyl-cyclohexyl)-methylsulfanyl]-acetyl}-12-[(4-aminomethyl-benzylamino)-methyl] mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 1 and 85, but using appropriate starting materials.

(166) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.69 (d, 2H, arom., J=7.6 Hz), 7.57 (d, 2H, arom., J=7.4 Hz), 5.47 (d, 1H, H-14, J=4.8 Hz), 5.34 (bs, 1H, 11-OH), 1.41 (s, 3H, H-15), 0.93 (s, 3H, H-18), 0.84 (bs, 3H, H-17), 0.66 (bs, 3H, H-16)

(167) MS m/e: 642 [M.sup.++H]

Preparation of the required Thioacetic acid S-(4-azidomethyl-cyclohexylmethyl) ester

(168) ##STR00228##

Step a: Thioacetic acid S-(4-methanesulfonyloxymethyl-cyclohexylmethyl) ester

(169) 5 g of Methanesulfonic acid 4-methanesulfonyloxymethyl-cyclohexylmethyl ester (described in JOC, 22, 994-5; 1957) was dissolved in 100 mL of DMF, 2.9 g of potassium thioacetate was added and the reaction mixture heated to 110 C. for 2 h. The reaction was quenched by pouring into water, extracted 3 with EtOAc, the organic phase washed with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness yielding a brown oil which was chromatographed on silica using cyclohexane/EtOAc=2:1 as eluent. The title compound was obtained as brown oil.

Step b: Thioacetic acid S-(4-azidomethyl-cyclohexylmethyl) ester

(170) 2.2 g of Thioacetic acid S-(4-methanesulfonyloxymethyl-cyclohexylmethyl) ester was dissolved in 100 mL of DMF, 2.1 g of sodium azide was added and the reaction mixture stirred at 70 C. for 16 h. The reaction mixture was poured into water, extracted 3 with EtOAc, the organic phase washed with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness yielding a brown oil which was used for the next step without further purification.

(171) According, e.g. analogously, to a method as set out under Example 99 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 9 are prepared. Chemical characterisation data are also set out in Table 9

(172) TABLE-US-00009 TABLE 9 Example R.sub.2EX R.sub.1EX 101 12-epi-12-desvinyl-14-O-{1-(2-Amino-acetyl)- piperidin-4-yl-sulfanyl]-acetyl}-12-[(4- aminomethyl-3-fluoro-benzylamino)-methyl] mutilin trihydrochloride MS m/e: 675 [M.sup.+ + H] 0 102 12-epi-12-Desvinyl-14-O-{{4-[(2-Amino- acetylamino)-methyl]-cyclohexylsulfanyl}- acetyl}-12-[(4-aminomethyl-3-fluoro- benzylamino)-methyl] mutilin tetrahydrochloride MS m/e: 703 [M.sup.+ + 2H] 103 12-epi-12-desvinyl-14-O-{[5-Aminomethyl- pyridin-2-yl-sulfanyl]-acetyl}-12-[(4- aminomethyl-2,5-difluoro-benzylamino)-methyl] mutilin tetrahydrochloride MS m/e: 659 [M.sup.+ + H] The required [4-(aminomethyl)-2,5-difluoro- phenyl]methanamine can be obtained byreaction of 1,4-bis(bromomethyl)-2,5-difluoro-benzene with excess NH.sub.3/THF at rt. The reductive amination was done as described in Example 1, step 4 using 6 eq of the diamine

(173) According, e.g. analogously, to a method as set out under Example 85 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 10 are prepared. Chemical characterisation data are also set out in Table 10

(174) TABLE-US-00010 TABLE 10 Example R.sub.2EX R.sub.1EX 104 12-epi-12-desvinyl-14-O-[(4-Aminomethyl- phenylsulfanyl)-acetyl]-12-[(2-amino-1-aminomethyl- ethylamino)-methyl] mutilin tetrahydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.45 (d, 2H, arom., J = 8 Hz), 7.36 (d, 2H, arom., J = 8 Hz), 5.47 (d, 1H, H-14, J = 7 Hz), 1.36 (s, 3H, H-15), 0.95 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 5.8), 0.59 (d, 3H, H-16, J = 5.6 Hz). [0256] MS m/e: 575 [M.sup.+ + H] The intermediate (2-Amino-3-tert- butoxycarbonylamino-propyl)-carbamic acid tert-butyl ester is e.g. described in ChemMedChem, 4(7), 1182- 1188; 2009 105 12-epi-12-Desvinyl-14-O-[(5-aminomethyl-pyridin-2- yl-sulfanyl)-acetyl]-12-[(6-amino-hexylamino)- methyl] mutilin tetrahydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.82-7.77 (m, 1H, arom.), 7.42-7.38 (m, 1H, arom.), 5.47 (bs, 2H, H- 14, 11-OH), 3.93 (s, 2H, H-22), 1.38 (s, 3H, H-15), 0.95 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6 Hz), 0.65 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 603 [M.sup.+ + H]

EXAMPLE 106

12-epi-12-desvinyl-14-O-{(4-[(2-Amino-acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride

Step 1: 12-epi-12-desvinyl-14-O-{(4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] motilin

(175) ##STR00239##

(176) A mixture of 0.3 mmol of BOC-Glycine, 0.45 mmol of HATU and 0.6 mmol of DIPEA in 15 mL of THF was stirred at 25 C. for 45 min. To the mixture obtained 0.3 mmol of 12-epi-12-desvinyl-14-O-[(4-Aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl]mutilin (Example 85, step 2) was added slowly. The mixture obtained was stirred at rt for another 2 h, quenched with water and extracted with EtOAc. The organic phase obtained was washed with water, brine, dried over Na.sub.2SO.sub.4, concentrated and the concentration residue was subjected to chromatography (DCM/MeOH=10:1) yielding the title compound.

(177) Step 2; 12-epi-12-desvinyl-14-O-{(4-[(7-Amino-acetylamino)-methyl]phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride BOC-deprotection of the compound obtained in step 1 was carried out as described in Example 1, Step 6.

(178) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.32 (d, 2H, arom., J=8 Hz), 7.25 (d, 2H, arom., J=8 Hz), 5.45 (d, 1H, H-14, J=5.4 Hz), 1.37 (s, 3H, H-15), 0.88 (s, 3H, H-18), 0.82 (d, 3H, H-17, J=5 Hz), 0.59 (d, 3H, H-16, J=5.2 Hz);

(179) MS m/e: 617 [M.sup.++H].

(180) ##STR00240##

(181) According, e.g. analogously, to a method as set out under Example 106 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 11 are prepared. Chemical characterisation data are also set out in Table 11.

(182) TABLE-US-00011 TABLE 11 Example R.sub.2EX R.sub.1EX 107 12-epi-12-desvinyl-14-O-{(4-[(2-Amino-3-(4- hydroxy-phenyl)-propionylamino)-methyl]- phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)- methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d6): 7.27 (d, 2H, arom., J = 8 Hz), 6.70 (d, 2H, arom., J = 8 Hz), 7.06-6.99 (m, 4H, arom.), 5.45 (d, 1H, H-14, J = 4.6 Hz), 1.37 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 4.6 Hz), 0.58 (d, 3H, H-16, J = 3 Hz) MS m/e: 723 [M.sup.+ + H] 108 12-epi-12-desvinyl-14-O-{(4-[(3-Amino- propionylamino)-methyl]-phenylsulfanyl)-acetyl}- 12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.32 (d, 2H, arom., J = 8 Hz), 7.22 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H- 14, J = 6.2 Hz), 1.37 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 5.2 Hz), 0.58 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 631 [M.sup.+ + H] 109 12-epi-12-desvinyl-14-O-{(4-[(2-Amino- acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-[4- aminomethyl-benzylamino-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.29 (d, 2H arom., J = 8 Hz), 7.21 (d, 2H arom., J = 8 Hz), 5.41 (bs, 2H, H- 14, 11-OH), 1.35 (s, 3H, H-15), 0.82 (bs, 6H, H-17, H- 18), 0.56 (d, 3H, H-16, J = 3.2 Hz) MS m/e: 679 [M.sup.+ + H] 110 12-epi-12-desvinyl-14-O-{(4-[(2-Amino- acetylamino)-methyl]-phenylsulfanyl)-acetyl}-12-(6- amino-hexylamino-methyl) mutilin trihydrochloride MS m/e: 659 [M.sup.+ + H]

EXAMPLE 111

12-epi-12-desvinyl-14-O-{[(3-Acetylamino-methyl)-phenylsulfanyl]-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin dihydrochloride

(183) ##STR00249##

(184) was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(185) MS m/e: 602 [M.sup.++H].

Preparation of the intermediate N-(3-Mercapto-benzyl)-acetamide

Step a: [3-(3-Hydroxymethyl-phenyldisulfanyl)-phenyl]-methanol

(186) 2 g of 3-mercaptobenzyl alcohol was dissolved in 20 mL of MeOH. 1.8 g of iodine was added in portions with cooling until the colour of iodine persisted. The mixture obtained was diluted with EtOAc, washed with brine; the organic phase was dried over Na.sub.2SO.sub.4, brought to dryness and the brown oil obtained was used without purification in the next step.

Step b: Methanesulfonic acid 3-(3-methanesulfonyloxymethyl-phenyldisulfanyl)-benzyl ester

(187) 2 g of [3-(3-hydroxymethyl-phenyldisulfanyl)-phenyl]-methanol was dissolved in 20 mL of THF, 2.7 mL of N-methylmorpholine was added followed by 3.2 g of methanesulfonic anhydride and stirring was maintained overnight at rt. The mixture obtained was diluted with EtOAc, washed with brine, dried over Na.sub.2SO.sub.4, evaporated to dryness and dried in high vacuum for 30 min. The brown oil obtained was used without purification in the next step.

Step c: 3-(3-Aminomethyl-phenyldisulfanyl)-benzylamine

(188) 3.1 g of methanesulfonic acid 3-(3-methanesulfonyloxymethyl-phenyldisulfanyl)-benzyl ester was taken up in 50 mL of MeOH/25% ammonia solution 1:1 and stirred at rt overnight. The mixture obtained was partitioned between brine and DCM; the organic phase obtained was dried over Na.sub.2SO.sub.4 and brought to dryness. The colourless crystals obtained were used without purification in the next step.

Step d: N-{3-[3-(Acetylamino-methyl)-phenyldisulfanyl]-benzyl}-acetamide

(189) 1.2 g of 3-(3-aminomethyl-phenyldisulfanyl)-benzylamine was dissolved in 20 mL of DCM; cooled to 0-5 C. and 3.1 mL of DIPEA and 1.7 mL of acetic anhydride were added. The mixture obtained was stirred at rt overnight, diluted with EtOAc and washed with 1N HCl. The aqueous phase obtained was washed with EtOAc, and the combined organic layers were treated with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The colourless crystals obtained were used without purification in the next step.

Step e: N-(3-Mercapto-benzyl)-acetamide

(190) 1.55 g of N-{3-[3-(acetylamino-methyl)-phenyldisulfanyl]-benzyl}-acetamide was dissolved in 20 mL of DMF and 860 mg of DTT was added and stirring was maintained for 2 h. The mixture obtained was partitioned between EtOAc and brine, the organic phase was dried over Na.sub.2SO.sub.4, the solvent was evaporated to dryness and the residue obtained was dried in high vacuum for 10 min. The title compound was obtained in the form of a colourless oil.

EXAMPLE 112

(191) 12-epi-12-desvinyl-14-O-{(4-{[2-Amino-3-(4-aminomethyl-phenyl)-propionylamino]-methyl}-phenylsulfanyl)-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride was obtained similarly and in analogy to a method as set out in Example 106, but using appropriate starting materials.

(192) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.44 (d, 2H arom., J=8 Hz), 7.28 (4H arom.), 7.12 (d, 2H arom. J=8 Hz), 5.46 (d, 1H, H-14, J=6.2 Hz), 1.37 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.81 (d, 3H, H-17, J=6.2 Hz), 0.58 (d, 3H, H-16, J=5.6 Hz); MS m/e: 736 [M.sup.++H].

(193) ##STR00250##

Preparation of the intermediate 2-tert-Butoxycarbonylamino-3-[4-(tert-butoxycarbonylamino-methyl)-phenyl]-propanoic acid

Step a: 2-Amino-3-(4-cyano-phenyl)-propanoic acid

(194) A mixture of 3.9 g of 4-bromomethyl-benzonitrile, 5.34 g of (benzhydrylidene-amino)-acetic acid ethyl ester, 522 mg of Bu.sub.4NBr and 5.16 g K.sub.2CO.sub.3 in 100 mL of CH.sub.3CN was stirred at 25 C. for 24 h. The mixture obtained was filtered and the filtrate obtained was concentrated. The concentration residue obtained was treated with 50 mL of 2N HCl and 50 mL THF within 12 h. The mixture obtained was extracted with EtOAc (50 mL*3) and the pH of the aqueous phase was adjusted to pH=8 with Na.sub.2CO.sub.3. The mixture obtained was extracted 3 with 50 mL EtOAc, the organic phase obtained was washed with water, brine, dried over Na.sub.2SO.sub.4 and concentrated. The crude product obtained was used without purification in the next step.

Step b: Ethyl 2-(tert-butoxycarbonylamino)-3-[4-[(tert-butoxycarbonylamino)-methyl]-phenyl]-propanoate

(195) A mixture of 3 g of 2-amino-3-(4-cyano-phenyl)-propanoic acid, 7.2 g of BOC.sub.2O and 500 mg of Pd/C in 100 mL of THF was stirred at rt for 16 h. The mixture obtained was filtered and the filtrate obtained was concentrated in vacuo. The crude product obtained was used without purification in the next step.

Step c: 2-tert-Butoxycarbonylamino-3-[4-(tert-butoxycarbonylamino-methyl)-phenyl]-propanoic acid

(196) A mixture of 6 g of ethyl 2-(tert-butoxycarbonylamino)-3-[4-[(tert-butoxycarbonylamino)-methyl]-phenyl]-propanoate and 1.1 g of NaOH in 40 mL of THF and 20 mL of water was stirred at rt for 16 h. From the mixture obtained THF was removed in vacuo and the residue was extracted 3 with 50 mL EtOAc. The aqueous phase's pH was adjusted to pH=8 with Na.sub.2CO.sub.3. The mixture obtained was extracted 3 with 50 mL EtOAc. The organic phase obtained was washed with water, brine, dried over Na.sub.2SO.sub.4 and concentrated yielding the title compound.

EXAMPLE 113

12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride

Step 1: 12-epi-12-desvinyl-14-O-[(4-Formyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(197) ##STR00251##

(198) A mixture 0.92 g of 12-epi-12-desvinyl-14-O-[(4-Hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl]mutilin (Example 57 prior to BOC-deprotection) and 1.18 g of Dess-Martin reagent in 40 mL of DCM was stirred at rt for 2 h, diluted with water and extracted with DCM. The organic phase obtained was dried over Na.sub.2SO.sub.4, evaporated to dryness and subjected to column chromatography (DCM/MeOH=100/5). After concentration the title compound was obtained in the form of a white foam.

Step 2: 12-epi-12-desvinyl-14-O-{{4-[(3-tert-Butoxycarbonylamino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(199) ##STR00252##

(200) A mixture of 12-epi-12-desvinyl-14-O-[(4-Formyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin (0.25 g) and (3-Amino-propyl)-carbamic acid tert-butyl ester (0.38 mmol) in dichloroethane (40 mL) was stirred at rt for 1 h. To the mixture obtained 161 mg of sodium triacetoxyborohydride was added and the mixture obtained was stirred at rt for 2 h, quenched with NaHCO.sub.3, and extracted with DCM. The organic phase obtained was dried over Na.sub.2SO.sub.4, evaporated to dryness and the evaporation residue obtained was subjected to column chromatography (DCM/MeOH/aq. NH.sub.3=10:1:0.1). After concentration the title compound was obtained in the form of a white foam.

Step 3: 12-epi-12-desvinyl-14-O-{{4-[(3-Amino-propylamino)-methyl]-phenylsulfanyl}-acetyl}-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride

(201) ##STR00253##

(202) BOC-deprotection of the compound obtained in step 2 was carried out as described in Example 1, Step 6.

(203) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.56 (d, 2H, arom., J=8 Hz), 7.37 (d, 2H, arom., J=8 Hz), 5.47 (d, 1H, H-14, J=7.2 Hz), 1.36 (s, 3H, H-15), 1.0 (s, 3H, H-18), 0.82 (d, 3H, H-17, J=4.8 Hz), 0.60 (d, 3H, H-16, J=4 Hz)

(204) MS m/e: 614 [M.sup.++H].

(205) According, e.g. analogously, to a method as set out under Example 113 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 12 are prepared. Chemical characterisation data are also set out in Table 12.

(206) TABLE-US-00012 TABLE 12 Example R.sub.2EX R.sub.1EX 114 12-epi-12-desvinyl-14-O-{{3-[(3-Amino- propylamino)-methyl]-phenylsulfanyl}-acetyl}- 12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride MS m/e: 617 [M.sup.+ + H] 115 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl- benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12- [(3-amino-propylamino)-methyl] mutilin tetrahydrochloride MS m/e: 679 [M.sup.+ + H] 116 12-epi-12-desvinyl-14-O-[(3-Allylaminomethyl- phenylsulfanyl)-acetyl]-12-[(3-amino- propylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.38 (s, 4H, arom.), 1.35 (s, 3H, H-15), 0.90 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.4 Hz), 0.59 (d, 3H, H-16, J = 4.6 Hz) MS m/e: 600 [M.sup.+ + H] 117 12-epi-12-desvinyl-14-O-{{4-[(3-Amino- propylamino)-methyl]-phenylsulfanyl}-acetyl}- 12-{[3-(3-amino-propoxy)-propylamino]-methyl} mutilin tetrahydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.56 (d, 2H arom., J = 8 Hz), 7.37 (d, 2H arom., J = 8 Hz), 5.46 (bs, 2H, H-14, 11-OH), 1.36 (s, 3H, H-15), 0.93 (s, 3H, H- 18), 0.82 (d, 3H, H-17, J = 6.2 Hz), 0.60 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 675 [M.sup.+ + H] 0 118 12-epi-12-desvinyl-14-O-[(4- Cyclopropylaminomethyl-phenylsulfanyl)-acetyl]- 12-[(3-amino-propylamino)-methyl] mutilin trihydrochloride MS m/e: 600 [M.sup.+ + H] 119 12-epi-12-desvinyl-14-O-[(4- Cyclopropylaminomethyl-phenylsulfanyl)-acetyl]- 12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.53 (d, 2H, arom., J = 8 Hz), 7.36 (d, 2H, arom., J = 8 Hz), 3.91 (s, 2H, H- 22), 1.37 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 6.4 Hz), 0.59 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 642 [M.sup.+ + H] 120 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl- benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12- [(6-amino-hexylamino)-methyl] mutilin tetrahydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.64-7.54 m (6H arom.), 7.37 (d, 2H arom., J = 8 Hz), 5.45 (bs, 2H, H- 14, 11-OH), 1.38 (s, 3H, H-15), 0.89 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.4 Hz), 0.60 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 721 [M.sup.+ + H] 121 12-epi-12-desvinyl-14-O-{{4-[(4-Aminomethyl- benzylamino)-methyl]-phenylsulfanyl}-acetyl}-12- [(4-aminomethyl-benzylamino)-methyl]- mutilin tetrahydrochloride MS m/e: 741 [M.sup.+ + H]

EXAMPLE 122

(207) 12-epi-12-desvinyl-14-O-[5-(3-Amino-propylcarbamoyl)-pyridin-2-ylsulfanyl]-acetyl-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(208) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.47 (d, 1H, arom., J=8.4 Hz), 5.47 (d, 1H, H-14, J=5.8 Hz), 4.01 (s, 2H, H-22), 1.37 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.81 (d, 3H, H-17, J=6 Hz), 0.64 (d, 3H, H-16, J=5.4 Hz)

(209) MS m/e: 674 [M.sup.++H].

(210) ##STR00270##

Preparation of the intermediate tert-Butyl N-[3-[(6-sulfanylpyridine-3-carbonyl)-amino]-propyl]-carbamate

Step a: 6-[(5-Chlorocarbonyl-2-pyridyl)-disulfanyl]-pyridine-3-carbonyl chloride

(211) 200 mg of 6,6-dithiodinicotinic acid was refluxed in 5 mL of thionyl chloride for 1 h and the solvent was removed azeotropically with CHCl.sub.3 on a rotary evaporator. The crude dichloride obtained was used immediately in the next step.

Step b: tert-Butyl N-[3-[[6-[[5-[3-(tert-butoxycarbonylamino)-propylcarbamoyl]-2-pyridyl]-disulfanyl]-pyridine-3-carbonyl]-amino]-propyl]-carbamate

(212) 222 mg of (3-Amino-propyl)-carbamic acid tert-butyl ester was dissolved in 10 mL of DCM, 445 l of TEA was added followed by dropwise addition of 220 mg of 6-[(5-chlorocarbonyl-2-pyridyl)-disulfanyl]-pyridine-3-carbonyl chloride in 5 mL of DCM with external cooling (ice bath). After 30 min the mixture obtained was diluted with water and brine and filtered biphasically. The filter residue was sucked reasonably dry and dried in high vacuum overnight yielding the title compound as a light brown powder.

Step c: tert-Butyl N-[3-[(6-sulfanylpyridine-3-carbonyl)-amino]-propyl]-carbamate

(213) 174 mg of tert-Butyl N-[3-[[6-[[5-[3-(tert-butoxycarbonylamino)-propylcarbamoyl]-2-pyridyl]-disulfanyl]-pyridine-3-carbonyl]-amino]-propyl]-carbamate and 58 mg of DTT were dissolved in 20 mL of DMF and the mixture obtained was stirred at rt for 30 min. The mixture obtained was diluted with EtOAc, washed with 1N HCl and water, dried over Na.sub.2SO.sub.4 and brought to dryness yielding the title compound in the form of a pale brown powder.

EXAMPLE 123

12-epi-12-desvinyl-14-O-[(2,5-Bis-aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride

Step 1: 12-epi-12-desvinyl-14-O-[(2,5-Bis-hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] mutilin

(214) ##STR00271##

(215) 81 mg of (4-hydroxymethyl-2-mercapto-phenyl)-methanol was dissolved in 4 mL of CH.sub.3CN, 53.4 mg of potassium tert-butoxide was added followed by 300 mg of 12-epi-12-desvinyl-14-O-[(toluene-4-yl-sulfonyloxy)-acetyl]-12-[(3-tert-butyloxycarbonylamino-propylamino)methyl] mutilin (Example 1 stop 1) in 2 mL of CH.sub.3CN and stirring for 20 min. To the mixture obtained 5 mg of potassium tert-butoxide and 7 mg of the thiol was added and stirring was continued for another 35 min where upon the reaction was deemed to be complete by HPLC determination. The mixture obtained was diluted with water and extracted 4 with EtOAc, the combined organic phases were washed with 2N NaOH, water, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The colourless foam obtained was used for the next step without further purification.

Step 2: 12-epi-12-desvinyl-14-O-[(2,5-Bis-azidomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] motilin

(216) ##STR00272##

(217) 295 mg of 12-epi-12-desvinyl-14-O-[(2,5-Bis-hydroxymethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl]mutilin was dissolved in 4.5 mL of THF, 267 mg of DPPA was added with external cooling, then 169 mg of DBU in 2 mL of CH.sub.3CN over 10 min. The cooling bath was removed and the mixture obtained was stirred at rt overnight. To the mixture obtained ca. 20 mL water was added, the resulting slurry was extracted with EtOAc (2), the combined organic phases were washed with water (2) and brine, dried over Na.sub.2SO.sub.4, brought to dryness and the dry residue obtained was subjected to chromatography (eluent: EtOAc/Et.sub.3N=100:1). The title compound was obtained in the form of colourless foamy crystals.

Step 3: 12-epi-12-desvinyl-14-O-[(2,5-Bis-aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl] motilin

(218) ##STR00273##

(219) 184 mg of 12-epi-12-desvinyl-14-O-[(2,5-Bis-azidomethyl-phenylsulfanyl)-acetyl]-12-[(3-tert-butoxycarbonylamino-propylamino)-methyl]mutilin was dissolved in 4 mL of EtOH, 1.35 mL of water was added and the mixture obtained was sonicated to obtain a clear solution. To the solution obtained 62.1 mg of NH.sub.4Cl and 43.4 mg of zinc powder were added and the mixture obtained was heated to reflux for 10 min and cooled to rt. To the mixture obtained 2 mL of 25% NH.sub.4OH was added and the phases were separated. The aqueous phase obtained was washed with EtOAc (2), the combined organic phases were dried, brought to dryness and the dry residue was subjected to chromatography over silica gel (eluent: EtOAc/EtOH/NH.sub.4OH=90:10:2). The title compound was obtained in the form of a colourless foam.

Step 4: 12-epi-12-desvinyl-14-O-[(2,5-Bis-aminomethyl-phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)-methyl] mutilin tetrahydrochloride

(220) ##STR00274##

(221) BOC-deprotection of the compound obtained in step 3 was carried out as described in Example 1, Step 6.

(222) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.77 (s, 1H, H-2, arom.), AB-system (v.sub.A=7.54, 2H, arom., v.sub.B=7.44, 2H, J=8 Hz), 5.43 (d, 1H, H-14, J=6.4 Hz), 1.34 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.81 (d, 3H, H-17, J=5.4 Hz), 0.55 (d, 3H, H-16, J=5.4 Hz)

(223) MS m/e: 623 [M.sup.+Cl]

Preparation of the intermediate (4-Hydroxymethyl-2-mercapto-phenyl)-methanol

Step a: 2-Chlorosulfonylterephthalic acid

(224) 15.3 g of 2-sulfoterephthalic acid sodium salt and 49 g of phosphorous pentachloride were mixed and the mixture was heated to 80 C. over night, cooled to rt and poured onto ice followed by extraction with diethyl ether. The organic phase obtained was washed successively with ice water until pH=4, and optionally with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The product was obtained in the form of colourless crystals.

(225) Step b: 2-Mercaptoterephthalic acid

(226) 19 g of 2-chlorosulfonylterephthalic acid was dissolved in 200 mL of THF and with external cooling 66 g of triphenylphosphine was added, followed by 1 mL of water and heating to reflux for 1 h. The mixture obtained was cooled to rt and poured onto 2N NaOH. The mixture obtained was washed twice with EtOAc; the aqueous phase obtained was acidified with 36% HCl, extracted with EtOAc, the organic phase obtained dried over Na.sub.2SO.sub.4 and evaporated to dryness yielding the title compound in the form of colourless crystals.

Step c: (4-Hydroxymethyl-2-mercapto-phenyl)-methanol

(227) 11.5 g (303 mmol) of LAH was slurried in 100 mL of dry THF and brought to reflux. To that mixture 4.4 g of 2-mercaptoterephthalic acid, dissolved in 100 mL of dry THF, was added slowly and after complete addition the mixture obtained was refluxed overnight. The mixture obtained was cooled in an ice bath and rendered acidic by the cautious addition of conc. HCl. The acidic mixture obtained was extracted with EtOAc, the organic phase obtained was stirred with solid NaHCO.sub.3, the mixture obtained was decanted, the organic phase obtained was dried over Na.sub.2SO.sub.4, evaporated to dryness and dried in high vacuum overnight yielding the title compound.

(228) According, e.g. analogously, to a method as set out under Example 123 above, but using appropriate starting materials, the compound of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 13 is prepared. Chemical characterisation data are also set out in Table 13.

(229) TABLE-US-00013 TABLE 13 Example R.sub.2EX R.sub.1EX 124 12-epi-12-desvinyl-14-O-[(3,5-Bis-aminomethyl- phenylsulfanyl)-acetyl]-12-[(3-amino-propylamino)- methyl] mutilin tetrahydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.58 (s, 2H, H-2, H-6, arom.), 7.37 (s, 1H, H-4, arom.), 5.48 (d, 1H, H-14, J = 6.2 Hz), 1.33 (s, 3H, H-15), 0.96 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5.6 Hz), 0.62 (d, 3H, H-16, J = 5 Hz) MS m/e: 589 [M.sup.+ + H]

EXAMPLE 125

12-epi-12-desvinyl-14-O-{[(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(2-guanidino-ethyl] mutilin dihydrochloride

Step 1: 4-epi-12-epi-12-(2-Amino-ethyl)-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin

(230) 305 mg of 4-epi-12-epi-12-(2-Allylamino-ethyl)-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin (prepared from the intermediate of step 4, Example 52, via reductive amination with allylamine in analogy to Example 1, step 4) in 10 mL of DCM was degassed in an ultrasonic bath for 10 min and then added to a mixture of 382 mg of N,N-dimethylbarbituric acid and 19 mg of tetrakis(triphenylphosphino) palladium (0) under an argon atmosphere. The mixture obtained was stirred at rt for 14 h. Since the reaction was not complete, the same amount of reagents was added and stirring was continued for another 24 h. The mixture obtained was diluted with DCM, washed with 10% K.sub.2CO.sub.3 solution (2), the combined aqueous phases were washed with DCM, the combined organic layers were dried over Na.sub.2SO.sub.4 and evaporated to dryness. The residue in the form of orange foamy crystals was subjected to silica gel chromatography (eluent: EtOAc/Et.sub.3N=100:1, 80:1, 60:1 and finally MeOH/Et.sub.3N=10:1). The title compound of formula

(231) ##STR00277##

(232) was obtained in the form of an orange-red semicrystalline oil.

Step 2: 4-epi-12-epi-12-(2-guanidino-ethyl)-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin

(233) ##STR00278##

(234) 192 mg of 4-epi-12-epi-12-(2-Amino-ethyl)-14-O-{{4-[(3-tert-butoxycarbonylamino-propylcarbamoyl)-methyl]-phenylsulfanyl}-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin and 75 mg of S-methylisothiuronium iodide was dissolved in 10 mL of THF and the mixture obtained was stirred for 14 h at rt. Then again 75 mg of isothiuronium iodide was added and the mixture obtained was stirred for another 4 d, diluted with EtOAc, washed with 5% NaHCO.sub.3 (2), dried over Na.sub.2SO.sub.4 and brought to dryness. The title compound was obtained in the form of pale orange crystals which were used without purification in the next step.

Step 3: 12-epi-12-desvinyl-14-O-{[(3-Amino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(2-guanidino-ethyl] mutilin dihydrochloride

(235) ##STR00279##

(236) 174 mg of 4-epi-3-Methoxy-12-epi-12-desvinyl-14-O-{[4-(3-tert-butoxycarbonylamino-propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-(2-guanidino-ethyl) mutilin was dissolved in 0.5 mL of DCM, 2 mL of 4N HCl in dioxane was added and after 15 min 5 mL of diethyl ether. The mixture obtained was stirred for 1 h and the formed precipitate was filtered off with suction, washed 5 with ether and dried in a rotovap bulb under vacuum without heating for 1 h. The crude product (brown crystals) was subjected to reverse chromatography (LiChroprep RP-18 (40-63 m)) with CH.sub.3CN/H.sub.2O=0-30%. The title compound was obtained in the form of colourless crystals after lyophilisation.

(237) MS m/e: 630 [M.sup.++H].

(238) According, e.g. analogously, to a method as set out under Examples 1 and 23 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 14 are prepared. Chemical characterisation data are also set out in Table 14.

(239) TABLE-US-00014 TABLE 14 Example R.sub.2EX R.sub.1EX 126 12-epi-12-desvinyl-14-O-{[4-(3-Hydroxy- propylcarbamoyl)-phenylsulfanyl]-acetyl}-12-[(6- amino-hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.57 (t, 1H, CONH, J = 6 Hz), 7.82 (d, 2H, arom., J = 8 Hz), 7.38 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H-14, J = 6.8 Hz), 5.38 (bs, 1H, 11-OH), 3.96 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.87 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.4 Hz), 0.59 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 674 [M.sup.+ + H] 0 127 12-epi-12-desvinyl-14-O-[(2-Hydroxy- ethylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)- methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H-14, J = 7.6 Hz), 1.41 (s, 3H, H-15), 0.99 (s, 3H, H-18), 0.83 (d, 3H, H-17, J = 6 Hz), 0.65 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 541 [M + H]

EXAMPLE 128

12-epi-12-desvinyl-14-O-{[3-(2,2-Difluoro-ethylamino)-cyclohexylsulfanyl]-acetyl}-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(240) ##STR00284##

(241) was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(242) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H-14, J=6.8 Hz), 1.40 (s, 3H, H-15), 0.98 (s, 3H, H-18), 0.82 (d, 3H, H-17, J=6 Hz), 0.65 (d, 3H, H-16, J=5 Hz)

(243) MS m/e: 658 [M+H].

Preparation of thioacetic acid S-[3-(2,2-difluoro-ethylamino)-cyclohexyl] ester

Step a: Thioacetic acid S-(3-oxocyclohexyl) ester

(244) A mixture of 2.45 mL of 2-cyclohexene-1-one, 1.96 mL of thioacetic acid and 11 mL of water was stirred vigorously over 5 h at rt, extracted with DCM (2); the combined organic phases were washed successively with 5% NaHCO.sub.3, 2N HCl and brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The evaporation residue obtained was subjected to chromatography over silica gel (eluent: cyclohexane/EtOAc=30:1). The title compound was obtained in the form of a pale yellow oil.

Step b: Thioacetic acid S-[3-(2,2-difluoroethylamino)cyclohexyl] ester

(245) 1 g of thioacetic acid S-(3-oxocyclohexyl) ester and 470 mg of 2,2-difluoroethylamine were dissolved with stirring in 20 mL of dichloroethane; 2.7 g of sodium triacetoxyborohydride and 498 L of acetic acid was added and the mixture obtained was stirred overnight at rt followed by neutralisation with 5% aqueous NaHCO.sub.3 solution and extraction with EtOAc (2). The combined organic phases obtained were dried over Na.sub.2SO.sub.4 and brought to dryness. The residue obtained was subjected to chromatography over silica gel (eluent: toluene/acetone=8:1) to obtain the title compound in the form of a yellow oil.

(246) According, e.g. analogously, to a method as set out under Example 1 above, but using appropriate starting materials, the compound of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 15 is prepared. Chemical characterisation data are also set out in Table 15.

(247) TABLE-US-00015 TABLE 15 Example R.sub.2EX R.sub.1EX 129 12-epi-12-desvinyl-14-O-[(2-Amino-7H-purin-6- ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.53 (s, 1H, arom.), 5.46 (d, 1H, H-14, J = 6.8 Hz), AB (2H, H-22, v.sub.A = 4.23, v.sub.B = 4.13, J = 16 Hz), 1.35 (s, 3H, H-15), 0.90 (s, 3H, H- 18), 0.79 (d, 3H, H-17, J = 6 Hz), 0.61 (d, 3H, H-16, J = 5 Hz). [0306] MS m/e: 630 [M + H]

EXAMPLE 130

12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(6-amino-hexylamino)-methyl] mutilin trihydrochloride

(248) ##STR00287##
was obtained similarly and in analogy to a method as set out in Example 1, but using appropriate starting materials.

(249) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.47 (d, 1H, H-14, J=7 Hz), 5.35 (bs, 1H, 11-OH), 1.40 (s, 3H, H-15), 0.97 (s, 3H, H-18), 0.82 (d, 3H, H-17, J=5.8 Hz), 0.64 (d, 3H, H-16, J=5.2 Hz).

(250) MS m/e: 594 [M+H]

Preparation of the intermediate tert-butyl 4-acetylsulfanyl-azepane-1-carboxylate

Step a: 4-Oxo-azepane-1-carboxylic acid tert-butyl ester

(251) 2.5 g of 4-Oxo-piperidine-1-carboxylic acid tert-butyl ester was dissolved in diethyl ether, cooled to 35 C., 2 mL of diazoacetic acid diethyl ester and 2.15 mL of borontrifluoride diethyl ether complex were added simultaneously and stirring at 35 C. was continued for one h. The mixture obtained was allowed to warm to rt and was rendered alkaline with 30% K.sub.2CO.sub.3 solution. The mixture obtained was extracted with EtOAc, the organic phase obtained was dried over Na.sub.2SO.sub.4, evaporated to dryness and the residue was dried in high vacuum leaving a yellow oil. The oil was taken up in 12 mL of THF, treated with 1.8 g of LiOH in 4 mL of water and the mixture obtained was refluxed overnight, cooled to rt and partitioned between brine and EtOAC. The phases obtained were separated and the organic phase obtained was dried over Na.sub.2SO.sub.4, brought to dryness and dried in high vacuum. The title compound was obtained in the form of colourless crystals.

Step b: 4-Hydroxy-azepane-1-carboxylic acid tert-butyl ester

(252) 2.06 g of 4-Oxo-azepane-1-carboxylic acid tert-butyl ester was dissolved in 20 mL of THF/MeOH 4:1, treated with external cooling with 370 mg of sodium borohydride and stirred with cooling for 1 h. The cold solution obtained was quenched with 10 mL of MeOH/H.sub.2O 1:1, the organic solvents were removed in vacuo and the residue obtained was partitioned between DCM and water; the organic phase was separated, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound in the form of a colourless oil was obtained.

Step c: tert-Butyl 4-methylsulfonyloxy-azepane-1-carboxylate

(253) 2.08 g of 4-Hydroxy-azepane-1-carboxylic acid tert-butyl ester, dissolved in 10 mL of DCM, was treated with external ice cooling with 2.02 g of methanesulfonic anhydride followed by 2.02 mL of TEA and stirred for one h. The phases obtained were separated, the organic phase obtained was washed with brine, dried over Na.sub.2SO.sub.4 and brought to dryness. The title compound in the form of a semicrystalline oil was obtained.

Step d: tert-Butyl 4-acetylsulfanyl-azepane-1-carboxylate

(254) 2.75 g of tert-Butyl 4-methylsulfonyloxy-azepane-1-carboxylate and 2.14 g of potassium thioacetate was dissolved in 10 mL of DMF and warmed to 70 C. for 3 h. From the mixture obtained the solvent was partially removed in vacuo, the residue partitioned between EtOAc/n-heptane=3:1 and water, the organic phase was separated, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The title compound in the form of a dark orange oil was obtained.

(255) According, e.g. analogously, to a method as set out under Examples 1 and 130 above, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 16 are prepared. Chemical characterisation data are also set out in Table 16.

(256) TABLE-US-00016 TABLE 16 Example R.sub.2EX R.sub.1EX 131 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-[(6-guanidino-hexylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.47 (d, 1H, H-14, J = 7.6 Hz), 5.37 (bs, 1H, 11-OH), 1.40 (s, 3H, H-15), 0.98 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 6.4 Hz), 0.64 (d, 3H, H-16, J = 4.8 Hz) MS m/e: 637 [M.sup.+ + H] 132 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-[(4-aminomethyl-benzylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.68 (d, 2H, arom., J = 8 Hz), 7,53 (d, 2H, arom., J = 8 Hz), 5.45 (d, 1H, H-14, J = 7.2 Hz), 5.30 (bs, 1H, 11-OH), 1.39 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.4 Hz), 0.63 (d, 3H, H-16, J = 5 Hz) MS m/e: 615 [M.sup.+ + H] 0 133 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-[(6-amino-octylamino)-methyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.45 (d, 1H, H-14, J = 5 Hz), 5.36 (bs, 1H, 11-OH), 1.39 (s, 3H, H-15), 0.96 (s, 3H, H-18), 0.81 (d, 3H, H-17, J = 5 Hz), 0.63 (d, 3H, H-16, J = 3.8 Hz) MS m/e: 622 [M.sup.+ + H] The reductive amination was done as described in Example 1, step 4 using 10 eq of 1,8-diaminooctane

(257) According, e.g. analogously, to a method as set out under Example 52, but with altered order of events (step 2.fwdarw.step 4.fwdarw.step 5.fwdarw.step 3.fwdarw.step 6, cf. Reaction Scheme 2) and using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 17 are prepared. Chemical characterisation data are also set out in Table 17.

(258) TABLE-US-00017 TABLE 17 Example R.sub.2EX R.sub.1EX 134 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12- [(6-amino-hexylamino)-ethyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.47 (d, 1H, H-14, J = 7.8 Hz), 1.40 (s, 3H, H-15), 0.98 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 5.8 Hz), 0.64 (d, 3H, H-16, J = 5 Hz) MS m/e: 608 [M.sup.+ + H] 135 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl)]-acetyl}-12- [(4-aminomethyl-benzylamino)-ethyl] mutilin triihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.61 (d, 2H, arom., J = 8 Hz), 7.52 (d, 2H, arom., J = 8 Hz), 5.48 (d, 1H, H-14, J = 5 Hz), 4.77 (bs, 1H, 11-OH), 1.38 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 6 Hz), 0.64 (d, 3H, H-17, J = 5.2 Hz), MS m/e: 671 [M.sup.+ + H] 136 12-epi-12-desvinyl-14-O-[5-Hydroxymethyl- pyridin-2-yl-sulfanylacetyl]-12-[(4- aminomethyl-3-fluoro-benzylamino)-ethyl] mutilin tetrahydrochloride MS m/e: 655 [M.sup.+ + H] 137 12-epi-12-desvinyl-14-O-{4-[(2-Amino- acetylamino)-cyclohexylsulfanyl]-acetyl}-12- [(4-aminomethyl-3-fluoro-benzylamino)- ethyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.66-7.53 (m, 2H, arom.), 7.44 (d, 1H, arom., J = 7.6 Hz, 5.48 (d, 1H, H-14, J = 7.2 Hz), 4.72 (d, 1H, 11-OH, J = 5.8 Hz), 1.38 (s, 3H, H-15), 1.37 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.83 (d, 2H, H-17, J = 6.4 Hz), 0.65 (d, 3H, H-16, J = 5.4 Hz) MS m/e: 703 [M.sup.+ + H] The required Thioacetic acid S-[4-(2-tert- butoxycarbonylamino-acetylamino)-cyclohexyl] ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials 00 01 138 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12- [(4-aminomethyl-3-fluoro-benzylamino)- ethyl] mutilin trihydrochloride MS m/e: 689 [M.sup.+ + H] 02 03 139 12-epi-12-desvinyl-14-O-[(5-Aminomethyl- pyridin-2-yl-sulfanyl)-acetyl]-12-[(4- aminomethyl-2,5-difluoro-benzylamino)- ethyl] mutilin tetrahydrochloride MS m/e: 673 [M.sup.+ + H] 04 05 140 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12- [(4-aminomethyl-2,5-difluoro-benzylamino)- ethyl] mutilin trihydrochloride MS m/e: 707 [M.sup.+ + H] 06 07 141 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12{2- [4-(2-amino-ethoxy)-benzylamino]-ethyl} mutilin trihydrochlorid MS m/e: 701 [M.sup.+ + H] 08 09 142 12-epi-12-desvinyl-14-O-{{4-[(2-Amino- acetylamino)-methyl]-cyclohexylsulfanyl}- acetyl}-12-[(4-aminomethyl-3-fluoro- benzylamino)-ethyl] mutilin trihydrochloride MS m/e: 717 [M.sup.+ + H] The required Thioacetic acid S-{4-[(2-tert- butoxycarbonylamino-acetylamino)-methyl]- cyclohexyl} ester was prepared in analogy to the procedure described in Example 78 using the appropriate starting materials

EXAMPLE 143

12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[(4-aminomethyl-phenylamino)-ethyl] mutilin trihydrochloride

Step 1: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(259) ##STR00310##

(260) was prepared by Dess-Martin oxidation from 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin (Example 52, step 2) in analogy to the procedure described in Example 52, step 4.

Step 2: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl]-12-(2-{4-[(2,2,2-trifluoro-acetylamino)-methyl]-phenylamino}-ethyl) mutilin

(261) ##STR00311##

(262) was prepared by reductive amination of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin with N-(4-Amino-benzyl)-2,2,2-trifluoro-acetamide according to the procedure described in Example 1, step 4.

Step 3: 12-epi-12-desvinyl-14-O-[(Toluene-4-sulfonyloxy)-acetyl]-12-(2-{4-[(2,2,2-trifluoro-acetylamino)-methyl]-phenylamino}-ethyl) motilin

(263) ##STR00312##

(264) 1.36 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl]-12-(2-{4-[(2,2,2-trifluoro-acetylamino)-methyl]-phenylamino}-ethyl) mutilin was dissolved in 10 mL of 1,4 dioxane and 2.5 mL of Lucas reagent was added. The mixture was stirred at ambient temperature for 2 h, diluted with EtOAc and washed with 2 water, NaHCO.sub.3 and brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The product was obtained as colorless foam and used for the next step without further purification.

Step 4: 12-epi-12-desvinyl-4-O-[(Azepan-4-ylsulfanyl)-acetyl]-12-[2-(4-aminomethyl-phenylamino)-ethyl] mutilin trihydrochloride

(265) ##STR00313##

(266) was prepared in analogy to Example 1, step 5 and 6 and Example 82 from 12-epi-2-desvinyl-14-O-[(toluene-4-sulfonyloxy)-acetyl]-12-(2-{4-[(2,2,2-Trifluoro-acetylamino)-methyl]-phenylamino}-ethyl) mutilin and Thioacetic acid S-azepan-4-yl ester. In order to ensure complete cleavage of the N-trifluoroacetyl group the reaction mixture was stirred an additional 20 min. at 40 C. The intermediate free amine was obtained as an off-white foam and converted into the hydrochloride salt without further purification.

(267) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.47 (m, 2H, arom.), 5.46 (d, 1H, H-14, J=6.6 Hz), 1.38 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.81 (d, 3H, H-17. J=6 Hz), 0.63 (d, 3H, H-16, J=5.2 Hz)

(268) MS m/e: 614 [M.sup.+Cl]

Preparation of the required N-(4-Amino-benzyl)-2,2,2-trifluoro-acetamide

(269) To 1 g of 4-(aminomethyl)-aniline in 50 mL of ethanol was slowly added 1.16 g of ethyl 2,2,2-trifluoroacetate at 4 C. The reaction mixture was allowed to warm to ambient temperature, diluted with EtOAc, washed with semi-saturated NaHCO.sub.3 solution and dried over Na.sub.2SO.sub.4. After evaporation the product was obtained as yellow crystals.

Preparation of the required Thioacetic acid S-azepan-4-yl ester

(270) Tert-Butyl 4-acetylsulfanyl-azepane-1-carboxylate (intermediate of Example 130, step d) was deprotected in analogy to Example 1, step 6 to give the title compound.

(271) According, e.g. analogously, to a method as set out under Example 143, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined, in Table 18 are prepared. Chemical characterisation data are also set out in Table 18.

(272) TABLE-US-00018 TABLE 18 Example R.sub.2EX R.sub.1EX 144 12-epi-12-desvinyl-14-O-{{4-[(2-Amino- acetylamino)]-cyclohexylsulfanyl}-acetyl}- 12-[(4-aminomethyl-phenylamino)-ethyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H-14, J = 4.6 Hz), 1.38 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 6.6 Hz), 0.64 (d, 3H, H-16, J = 5 Hz) MS m/e: 705 [M.sup. + Cl] Thioacetic acid S-[4-(2-amino-acetylamino)- cyclohexyl] ester (prepared byreaction of the corresponding intermediate of Example 137 with TFA in DCM) was used as intermediate in step 4 145 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)- piperidin-4-yl-methylsulfanyl]-acetyl}-12-[(4- aminomethyl-phenylamino)-ethyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.49 (d, 1H, H-14, J = 7.6 Hz), 1.38 (s, 3H, H-15), 0.92 (s, 3H, H-18), 0.82 (d, 3H, H-17, J = 6.6 Hz), 0.65 (d, 3H, H-16, J = 4.4 Hz) MS m/e: 705 [M.sup. + Cl] Thioacetic acid S-[1-(2-amino-acetyl)-piperidin- 4-ylmethyl] ester (prepared byreaction of the corresponding intermediate of Example 81 with TFA in DCM) was used as intermediate in step 4

EXAMPLE 146

12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-2-(8-amino-octyl) mutilin dihydrochloride

Step 1; 4-epi-12-epi-12-(8-Azido-oct-2-enyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin

(273) ##STR00318##

(274) 5.95 g of 6-Azidohexyl-triphenylphosphonium iodide was dissolved in 90 mL THF, 25 mL of 0.5M potassium hexamethyldisilazide in THF was added dropwise under argon at 78 C. After stirring at this temperature for 30 minutes 4.47 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-triethylsilyl mutilin in 20 mL THF was added dropwise, the reaction mixture was left in the cooling bath and allowed to warm to ambient temperature overnight. The suspension was poured into 500 mL water/20 mL 2N HCl, extracted 3 with EtOAc, the combined organic layers were washed with 5% sodium bicarbonate solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness, leaving a beige semi crystalline residue. Purification over silica gel (toluene/EtOAc=20:1) yielded the product as pale yellow oil.

Step 2: 4-epi-12-epi-12-(8-Amino-oct-2-enyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin

(275) ##STR00319##

(276) 4.31 g of 4-epi-12-epi-12-(8-Azido-oct-2-enyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin was hydrogenated in an H-cube apparatus at 50 bar over 10% Pd/C (flow 1 mL/min). After evaporation of the solvent the title compound was isolated.

Step 3: 4-epi-12-epi-12-(8-Amino-octyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin

(277) ##STR00320##

(278) 2.9 g (5.29 mmol) of 4-epi-12-epi-12-(8-Amino-oct-2-enyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin was dissolved in 200 mL THF/MeOH 1+1 and hydrogenated (balloon) over 290 mg of 10% Pd/C for 16 hours. The catalyst was filtered off and the solvent evaporated yielding the product as yellow oil containing some residual solvent.

Step 4: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin

(279) ##STR00321##

(280) 2.88 g of 4-epi-12-epi-12-(8-amino-octyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl mutilin and 637 mg of triethylamine were dissolved in 25 mL dichloromethane, cooled in an ice bath, treated dropwise with 1.15 g of trifluoroacetic acid anhydride in 15 mL dichloromethane and then stirred at ambient temperature for 1 hour. The reaction mixture was washed with 1N HCl (20 mL), 5% sodium bicarbonate solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The product was obtained as yellow oil.

Step 5: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin

(281) ##STR00322##

(282) 3.26 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-triethylsilyl-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin was suspended in 65 mL ethanol containing 2 wt. % hydrochloric acid (aq) and stirred at ambient temperature. After 3 hours the mixture was partitioned between 5% sodium bicarbonate solution and EtOAc, the aqueous phase washed with EtOAc, the combined organic layers were dried over Na.sub.2SO.sub.4 and evaporated to dryness to give a pale yellow oil which was chromatographed over silica gel (DCM/tert-butyl methyl ether=40:1) to give a colorless oil.

Step 6: 4-epi-12-epi-14-O-Bromoacetyl-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin

(283) ##STR00323##

(284) 750 mg (1.41 mmol) of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin was dissolved in 10 mL EtOAc, treated with 571 mg of triethylamine, 52 mg of 4-dimethylaminopyridine and warmed with stirring to 50 C. 854 mg of bromoacetic acid bromide in 10 mL EtOAc was added dropwise over 20 minutes, during which time the reaction mixture turned dark brown. After 15 minutes 0.79 mL of Et.sub.3N followed by 854 mg of bromoacetic acid bromide in 5 mL EtOAc and after further 15 minutes the same amount of Et.sub.3N and bromoacetic acid bromide was added after which no starting material could be detected by TLC (toluene/EtOAc 5:1). 1 mL methanol was added to destroy residual acid bromide and stirring was continued for 45 minutes. Then the reaction solution was shaken with 1N HClmL, filtered over celite biphasically, the organic layer washed with 5% sodium bicarbonate solution, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The crude product (black oil) was purified by chromatography over silica gel (eluant: DCM) and yielded the title compound as yellow oil.

Step 7: 4-epi-12-epi-14-O-{[1-(2-tert-Butoxycarbonylamino-acetyl)-piperidin-4-ylsulfanyl]-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin

(285) ##STR00324##

(286) 320 mg of 4-epi-12-epi-14-O-Bromoacetyl-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin and 233 mg of Thioacetic acid S-[1-(2-tert-butoxycarbonylamino-acetyl)-piperidin-4-yl] ester was dissolved in 10 mL methanol and treated with 294 l 5M aqueous potassium carbonate solution, stirred 15 minutes at ambient temperature and 30 minutes in an ultrasonic bath. The reaction mixture was diluted with EtOAc, washed with 1N HCl, 2N NaOH, 5% sodium bicarbonate solution, dried over Na.sub.2SO.sub.4 and brought to dryness; the crude product was obtained as an orange oil.

Step 8: 4-epi-12-epi-12-(8-Amino-octyl)-14-O-{[1-(2-tert-Butoxycarbonylamino-acetyl)-piperidin-4-ylsulfanyl]-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin

(287) ##STR00325##

(288) 134 mg of 4-epi-12-epi-14-O-{[1-(2-tert-Butoxycarbonylamino-acetyl)-piperidin-4-ylsulfanyl]-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-[8-(2,2,2-trifluoro-acetylamino)-octyl] mutilin was dissolved in 10 mL methanol, treated with 1.6 mL 1N sodium hydroxide solution and stirred at ambient temperature. After 1 hour 1.6 mL 1N sodium hydroxide solution was added and stirring was continued for 1 h. The solution was diluted with EtOAc and washed 2 with water; the combined aqueous layers were washed with EtOAc, the organic phases dried over Na.sub.2SO.sub.4 and evaporated to dryness to give a light yellow oil containing some residual solvent.

Step 9: 12-epi-12-desvinyl-14-O-{[1-(2-Amino-acetyl)-piperidin-4-ylsulfanyl]-acetyl}-12-(8-amino-octyl)-mutilin dihydrochloride

(289) ##STR00326##

(290) 129 mg of 4-epi-12-epi-12-(8-Amino-octyl)-14-O-{[1-(2-tert-Butoxycarbonylamino-acetyl)-piperidin-4-ylsulfanyl]-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin was dissolved in 0.5 mL dioxane, treated with 2 mL 4N HCl in dioxane and the homogeneous solution was stirred at ambient temperature overnight. Then the volatiles were removed in vacuo and the residue (yellow oil) was purified by reversed phase chromatography (eluant: acetonitrile in water 0-35%). The title compound was obtained as colourless crystals.

(291) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.46 (d, 1H, H-14, J=7.4 Hz), 1.33 (s, 3H, H-15), 0.83-0.77 (m, 6H, H-18, H-17), 0.62 (d, 3H, H-16, J=5.4 Hz)

(292) MS m/e: 636 [M.sup.++H]

Preparation of the required 6-Azidohexyl-triphenylphosphonium iodide

Step a: 6-Hydroxyhexyl-triphenylphosphonium iodide

(293) 7.97 g of 6-iodohexanol and 9.17 g of triphenylphosphine was dissolved in 10 mL of acetonitrile and refluxed for 20 hours. The mixture was cooled to ambient temperature, treated with 150 mL diethyl ether, stirred for about 4 hours and left standing overnight. The resulting precipitate was filtered off, washed five times with diethyl ether and dried in high vacuum overnight; the product was obtained as colorless powder.

Step b: 6-Bromohexyl-triphenylphosphonium iodide

(294) 7.97 g of 6-iodohexanol and 9.17 g of triphenylphosphine was dissolved in 10 mL of acetonitrile and refluxed for 20 hours. The mixture was cooled to ambient temperature, treated with 150 mL diethyl ether, stirred for about 4 hours and left standing overnight. The resulting precipitate was filtered off, washed five times with diethyl ether and dried in high vacuum overnight; the product was obtained as colorless powder.

Step c: 6-Azidohexyl-triphenylphosphonium iod

(295) 10.4 g of 6-Bromohexyl-triphenylphosphonium iodide was dissolved in 25 mL ethanol, 1.83 g of sodium azide in 25 mL water was added and the mixture was refluxed for ca. 16 hours. The volatiles were almost completely removed and the residue was stirred with 50 mL dichloromethane and 20 mL water for 30 minutes. After phase separation the aqueous layer was extracted with 3 dichloromethane, the combined organic layers were dried over Na.sub.2SO.sub.4 and brought to dryness yielding the title compound as a light brown resin

Preparation of the required 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-triethylsilyl mutilin Step a: 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo mutilin

(296) 53 g of Sodium hydroxide was dissolved in 500 mL of methanol with boiling. The temperature was lowered by 5 C. and then 240 g of 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl]mutilin was charged and the mixture was refluxed for 1 hour. Almost 90% of the solvent was removed in vacuo, the residue was diluted with external cooling with water, and then extracted with EtOAc. The combined organic phases were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound as yellow solid.

Step b: 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo-14-O-triethylsilyl mutilin

140 g of 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo mutilin

(297) 57 g of imidazole and 26 g of DMAP was dissolved in 2 L of DCM, then 126 g of Et.sub.3SiCl was added and the mixture stirred at rt overnight. The reaction was quenched with water, extracted with EtOAc, the organic phases were combined, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by chromatography (PE/EtOAc=10/1) to give the title compound as yellow solid.

Step c: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo-14-O-triethylsilyl mutilin

(298) 140 g of 4-epi-12-epi-3-deoxo-11-deoxy-3-methoxy-11-oxo-14-O-triethylsilyl mutilin was dissolved in 1 L of THF, then 1.25 L of 9-BBN (0.5 m in THF) was added and the reaction refluxed for 2 h. The reaction is cooled in an ice-bath, slowly 350 g of H.sub.2O.sub.2 (30%) and 310 mL of 2N NaOH) were added and stirred for 30 min. After warming to room temperature water and EtOAc were added, the organic phase was washed with brine, dried over Na.sub.2SO.sub.4, concentrated and the residue was purified by chromatography (PE/EtOAc=15/1) to give the title compound as white solid.

Step d: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-triethylsilyl mutilin

(299) 70 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(2-hydroxy-ethyl)-3-methoxy-11-oxo-14-O-triethylsilyl mutilin was dissolved in 1 L of DCM, then 64 g of Dess-Martin reagent was added and the reaction stirred at rt.

(300) After 2 h the reaction was quenched by addition of sat. NaHCO3 solution, the organic phase was separated, dried over Na.sub.2SO.sub.4, concentrated and the residue was purified by chromatography (PE/EtOAc=8/1) to give the title compound as white solid

(301) According, e.g. analogously, to a method as set out under Example 1 and 146, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 19 are prepared. Chemical characterisation data are also set out in Table 19.

(302) TABLE-US-00019 TABLE 19 Example R.sub.2EX R.sub.1EX 147 12-epi-12-desvinyl-14-O-{[1-(2-Amino- acetyl)-piperidin-4-yl-sulfanyl]-acetyl}-12-[3- (4-aminomethyl-phenyl)-propyl] mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.39 (d, 2H, arom., J = 8 Hz), 7.21 (d, 2H, arom., J = 8 Hz), 5.47 (d, 1H, H-14, J = 7 Hz), 4.48 (bs, 1H, 11- OH), 1.35 (s, 3H, H-15), 0.84 (m, 6H, H-18, H- 17), 0.63 (d, 3H, H-16, J = 5.2 Hz) MS m/e: 656 [M.sup.+ + H] Azidomethyl-phenyl-methyl-triphosphonium bromide was used as intermediate in step 1 148 12-epi-12-desvinyl-14-O-[(Azepan-4-yl- sulfanyl)-acetyl]-12-[3-(4-aminomethyl-phenyl)- propyl] mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.38 (d, 2H, arom., J = 8 Hz), 7.22 (d, 2H, arom., J = 8 Hz), 5.46 (d, 1H, H-14, J = 5.6 Hz), 3.96 (bs, 1H, 11- OH), 1.35 (s, 3H, H-15), 0.81 (m, 6H, H-18, H- 17), 0.62 (d, 3H, H-16, J = 4.2 Hz) MS m/e: 613 [M.sup.+ + H] 0 149 12-epi-12-desvinyl-14-O-[(Azepan-4-yl- sulfanyl)-acetyl]-12-(6-amino-hexyl) mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H-14, J = 7.6 Hz), 4.33 (d, 1H, 11-OH, J = 5.8 Hz), 1.36 (s, 3H, H-15), 0.84-0.79 (m, 4H, H- 18, H-17), 0.64 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 565 [M.sup.+ + H] 150 12-epi-12-desvinyl-14-O-[(Azepan-4-yl- sulfanyl)-acetyl]-12-(8-amino-octyl) mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.48 (d, 1H, H-14, J = 7.6 Hz), 4.40 (d, 1H, 11-OH, J = 5.38 Hz), 3.88 (s, 2H, H-22), 1.36 (s, 3H, H-15), 0.84-0.79 (m, 6H, H-18, H-17), 0.64 (d, 3H, H-16, J = 5.6 Hz) MS m/e: 593 [M.sup.+ + Cl]

EXAMPLE 151

12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethenyl) mutilin trihydrochloride

Step 1: 12-epi-14-O-{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl mutilin

(303) ##STR00335##

(304) 2.5 g of 12-epi-Pleuromutilintosylate was coupled with thioacetic acid S-{4-[(2-tert-butoxycarbonylamino-acetylamino)-methyl]-cyclohexyl} ester (Example 84) following the general procedure as described in Example 1, step 5. The title compound was obtained as colorless foam.

Step 2: 12-epi-12-desvinyl-14-O-{{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[(E)-2-(4-{[tert-butoxycarbonyl-(2-tert-butoxycarbonylamino-ethyl)-amino]-methyl}-phenyl)-ethenyl] mutilin

(305) ##STR00336##

(306) 3.5 g of 12-epi-14-O-{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl mutilin was dissolved in 20 mL of DMA, 5 g of (2-tert-Butoxycarbonylamino-ethyl)-(4-iodo-phenyl)-carbamic acid tert-butyl ester, 1.1 g of NMM and 223 mg of bis-(benzonitrile)-palladium(II)-chloride was added and the reaction mixture stirred at 100 C. for 20 h in a microwave synthesizer (Biotage). The reaction mixture was poured into water, extracted with EtOAc, washed with brine and water and concentrated to give the title product as colorless foam.

Step 3: 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethenyl) mutilin trihydrochloride

(307) The title compound was obtained following the general procedure as described in Example 1, step 6. The product was purified by reversed phase chromatography (eluant: acetonitrile in water 0-35%).

(308) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.39 (d, 2H, arom., J=8 Hz), 7.49 (d, 2H, arom., J=8 Hz), 6.55 (d, 2H, J=16 Hz), 6.30 (d, 2H, J=17 Hz), 5.57 (d, 1H, H-14, J=7 Hz), 1.41 (s, 3H, H-15), 1.19 (s, 3H, H-18), 0.88 (bs, 6H, H-17), 0.68 (bs, 3H, H-16)

(309) ##STR00337##

(310) MS m/e: 711 [M.sup.++H]

EXAMPLE 152

12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethyl) mutilin trihydrochloride

Step 1: 12-epi-12-desvinyl-14-O-{{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[(E)-2-(4-{[tert-butoxycarbonyl-(2-tert-butoxycarbonylamino-ethyl)-amino]-methyl}-phenyl)-ethyl] mutilin

(311) ##STR00338##

(312) 2 g of 12-epi-12-desvinyl-14-O-{{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[(E)-2-(4-{[tert-butoxycarbonyl-(2-tert-butoxycarbonylamino-ethyl)-amino]-methyl}-phenyl)-ethenyl] mutilin was dissolved in 1250 mL of MeOH and hydrogenated in an H-Cube apparatus using 10% Pd/C as catalyst (50 C., 1 mL/min, 60 bar). After concentration the product was obtained as yellow foam.

Step 2: 12-epi-12-desvinyl-14-O-{{4-[(2-Amino-acetylamino)-methyl]-cyclohexylsulfanyl}-acetyl}-12-[2-{4-[(2-amino-ethylamino)-methyl]-phenyl}-ethyl) mutilin trihydrochloride

(313) ##STR00339##

(314) The product was obtained following the general procedure as described in Example 1, step 6. The product was purified by reversed phase chromatography (eluant: acetonitrile in water 0-35%).

(315) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.45 (m, 2H, arom.), 7.22 (m, 2H, arom.), 5.51 (d, 1H, H-14, J=5.8 Hz), 1.40 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.85 (bs, 3H, H-17), 0.66 (bs, 3H, H-16)

(316) MS m/e: 713 [M.sup.++H]

Preparation of the required (2-tert-Butoxycarbonylamino-ethyl)-(4-iodo-phenyl)-carbamic acid tert-butyl ester

Step a [2-(4-Iodo-phenylamino)-ethyl]-carbamic acid tert-butyl ester

(317) To 10.5 g of 4-Iodobenzyl bromide in 100 mL of THF was added 11.4 g of (2-amino-ethyl)-carbamic acid tert-butyl ester and the reaction mixture was stirred for 12 h at rt. The reaction mixture was poured into water, extracted with EtOAc, washed with brine and water, dried over Na.sub.2SO.sub.4, concentrated and chromatographed on silica using DCM/MeOH=30:1. The title compound was obtained as yellow oil.

Step b: (2-tert-Butoxycarbonylamino-ethyl)-(4-iodo-phenyl)-carbamic acid tert-butyl ester

(318) 8.6 g of [2-(4-Iodo-phenylamino)-ethyl]-carbamic acid tert-butyl ester was dissolved in 200 mL of DCM, 5.4 g of BOC.sub.2O and 4.81 g of TEA was added and the reaction mixture stirred at rt for 16 h. The reaction mixture was poured into water, extracted with EtOAc, washed with 1n HCl and water. After concentration the title compound was obtained as colorless crystals.

(319) According, e.g. analogously, to a method as set out under Example 1, 151 and 152, but using appropriate starting materials, the compounds of formula I.sub.EX, wherein R.sub.1EX and R.sub.2EX are as defined in Table 20 are prepared. Chemical characterisation data are also set out in Table 20.

(320) TABLE-US-00020 TABLE 20 Example R.sub.2EX R.sub.1EX 153 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-[2-(4-Aminomethyl-phenyl)-ethyl]- mutilin dihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.41 (bs, 2H, arom.), 7.23 (bs, 2H, arom.), 5.52 (bs, 1H, H-14), 4.63 (bs, 1H, 11-OH), 1.40 (s, 3H, H-15), 0.91 (s, 3H, H-18), 0.84 (bs, 3H, H-17), 0.65 (bs, 3H, H- 16) MS m/e: 599 [M.sup.+ + H] 1-Azidomethyl-4-iodo-benzene (prepared from 1- Bromomethyl-4-iodo-benzene) was used as intermediate in step 1 0 154 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-((E)-2-pyridin-3-yl-ethenyl) mutilin hydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.88 (m, 1H, arom.), 8.67 (m, 1H, arom.), 8.45 (m, 1H, arom.), 7.91 (m, 1H, arom.), 6.81 (d, 2H, J = 17 Hz), 6.48 (d, 2H, J = 17 Hz), 5.52 (d, 1H, H-14, J = 7.4 Hz), 1.38 (s, 3H, H-15), 1.17 (s, 3H, H-18), 0.83 (d, 3H, H-17, J = 6.6 Hz), 0.65 (d, 3H, H-16, J = 4.6 Hz), MS m/e: 569 [M.sup.+ + H] 3-Bromo-pyridine was used as intermediate in step 1 155 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-((E)-2-{4-[(2-Amino-ethylamino)- methyl]-phenyl}-ethenyl) mutilin trihydrochloride .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.5 (d, 2H, arom., J = 8.2 Hz), 7.42 (d, 2H, arom., J = 8.2 Hz), 6.52 (d, 2H, J = 16.4 Hz), 6.29 (d, 2H, J = 16.4 Hz), 5.54 (d, 1H, H-14, J = 7.2 Hz), 4.46 (bs, 1H, 11- OH), 1.39 (s, 3H, H-15), 1.16 (s, 6H, H-18), 0.84 (d, 3H, H-16, J = 5.8 Hz), 0.67 (bs, 3H, H-16) MS m/e: 640 [M.sup.+ + H] 156 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-[2-{4-[(2-amino-ethylamino)-methyl]- phenyl}-ethyl) mutilin trihydrochloride MS m/e: 642 [M.sup.+ + H] 157 12-epi-12-desvinyl-14-O-[(Azepan-4-ylsulfanyl)- acetyl]-12-((E)-2-{4-[(2-amino-ethylamino)- methyl]-3-fluoro-phenyl}-ethenyl) mutilin trihydrochlorid .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 7.68-7.6 (m, 1H, arom.), 7.34-7.26 (m, 2H, arom.), AB (2H, CHCH, v.sub.A = 6.59, v.sub.B = 6.30, J = 16 Hz), 5.53 (d, 1H, H-14, J = 5.2 Hz), 1.39 (s, 3H, H-15), 1.08 (s, 3H, H-18), 0.84 (d, 3H, H-17, J = 6.4 Hz), 0.66 (d, 3H, H-16, J = 4.4 Hz) MS m/e: 658 [M.sup.+ + H] The required 1 Bromomethyl 2 fluoro 4 iodo benzene is described in: PCT Int. Appl., 2011132051, 27 Oct. 2011 158 12-epi-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl- sulfanyl]-acetyl}-12-((E)-2-{4-[(2-amino- ethylamino)-methyl]-phenyl}-ethenyl) mutilin trihydrochloride MS m/e: 683 [M.sup.+ + H] 0 159 12-epi-14-O-{[1-(2-Amino-acetyl)-piperidin-4-yl- sulfanyl]-acetyl}-12-[2-{4-[(2-amino-ethylamino)- methyl]-phenyl}-ethyl) mutilin trihydrochloride MS m/e: 685 [M.sup.+ + H]

EXAMPLE 160

12-epi-12-desvinyl-14-O-[(5-Aminomethyl-pyridin-2-ylsulfanyl)-acetyl]-12-[2-(4-aminomethyl-benzoylamino)-ethyl] mutilin trihydrochloride

Step 1: 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(hydroxyimino-ethyl)-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(321) ##STR00354##

(322) 617 mg of NH.sub.2OH.HCl and 355 mg of NaOH was dissolved in 60 mL MeOH, the mixture was stirred at 25 C. for 1 h, then 5.0 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-12-(2-oxo-ethyl)-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin (Example 143, step 1) was added and the mixture stirred at 25 C. for 12 hours. The mixture was concentrated and the residue purified by chromatography (PE/EtOAc=3:1) to give the title compound as colorless oil.

Step 2: 4-epi-12-epi-12-(2-Amino-ethyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(323) ##STR00355##

(324) 2.9 g of 4-epi-12-epi-3-deoxo-11-deoxy-12-desvinyl-12-(hydroxyimino-ethyl)-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin was dissolved in 50 mL of MeOH, then 672 mg of NiCl.sub.2 and 196 mg of NaBH.sub.4 was added at 78 C., afterwards stirred at 78 C. for 2 h, warmed to 25 C. and stirred for 2 h. The reaction was filtered, concentrated, the residue was dissolved in DCM, washed with water, the organic phase was separated, dried over Na.sub.2SO.sub.4, concentrated to give the title compound as colorless oil.

Step 3: 4-epi-12-epi-12-{2-[4-(tert-Butoxycarbonylamino-methyl)-benzoylamino]-ethyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin

(325) ##STR00356##

(326) 440 mg of 4-epi-12-epi-12-(2-Amino-ethyl)-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo-14-O-[(toluene-4-sulfonyloxy)-acetyl] mutilin was dissolved in 10 mL of DCM, then 240 mg of 4-(tert-Butoxycarbonylamino-methyl) benzoic acid, 364 mg of HATU and 242 mg of TEA was added and the mixture was stirred at 25 C. for 12 hours. Then the mixture was quenched with water, extracted with DCM, the organic phase was separated, dried over Na.sub.2SO.sub.4, concentrated and the residue was purified by chromatography (PE/EtOAc=3:1) to give the title compound as white solid.

Step 4: 4-epi-12-epi-12-{2-[4-(tert-Butoxycarbonylamino-methyl)-benzoylamino]-ethyl}-14-O-{[5-(tert-Butoxycarbonylamino-methyl)-pyridin-2-ylsulfanyl]-acetyl}-3-deoxo-11-deoxy-12-desvinyl-3-methoxy-11-oxo mutilin

(327) ##STR00357##

(328) was prepared following the general procedure as described in Example 1, step 5. After chromatography (PE/EtOAc=2:1) the title compound was obtained as white solid.

Step 5: 12-epi-12-desvinyl-14-O-[(5-Aminomethyl-pyridin-2-ylsulfanyl)-acetyl]-12-[2-(4-aminomethyl-benzoylamino)-ethyl] mutilin trihydrochloride

(329) ##STR00358##

(330) The product was obtained following the general procedure as described in Example 1, step 6.

(331) MS m/e: 651 [M.sup.++H]

(332) Comparator Compounds

(333) According to a method as described in Monatshefte fr Chemie 117, 1073 (1986) and a method as described in the examples, in particular in Examples 1 and 62 above, but using appropriate starting materials, compounds of formula IV are obtained

(334) ##STR00359##

(335) wherein R.sub.EX2C and the substituent and its stereospecity at position C-12 of the mutilin ring are as defined in table 21 below.

(336) TABLE-US-00021 TABLE 21 C-12 Example R.sub.2EXC substitution C1 12-epi-14-O-{[(5-Amino-4H-1,2,4-triazol-3-yl)- sulfanyl]-acetyl} mutilin hydrochloride (comparator compound) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 5.91 (dd, 1H, H-19, J.sub.E = 18 Hz, J.sub.Z = 12 Hz), 5.48 (d, 1H, H-14, J = 7.6 Hz), 4.96-4.84 (m, 2H, H-20), AB (2H, H-22, v.sub.A = 3.99, v.sub.B = 3.89, J = 16 Hz), 3.46 (d, 1H, H-11, J = 5.4 Hz), 1.32 (s, 3H, H-15), 0.99 (s, 3H, H-18), 0.80 (d, 3H, H-17, J = 6.6 Hz), 0.59 (d, 3H, H-16, J = 6 Hz) MS m/e: 477 [M.sup.+ + H] 0 C2 14-O-{{4-[(2-Amino-propylcarbamoyl)-methyl]- phenylsulfanyl}-acetyl} mutilin hydrochloride (comparator compound) .sup.1H-NMR (200 MHz, DMSO-d.sub.6): 8.35 (t, 1H, CONH, J = 6 Hz), 8.01 (bs, 3H, NH.sub.3.sup.+), 7.27 (d, 2H, arom., J = 8 Hz), 7.18 (d, 2H, arom., J = 8 Hz), 6.04 (dd, 1H, H- 19, J.sub.E = 18 Hz, J.sub.Z = 10 Hz), 5.50 (d, 1H, H-14, J = 7.6 Hz), 5.09-4.92 (m, 2H, H-20), AB (2H, H-22, v.sub.A = 3.82, v.sub.B = 3.72, J = 16 Hz), 1.31 (s, 3H, H-15), 0.99 (s, 3H, H-18), 0.79 (d, 3H, H-17, J = 6.8 Hz), 0.55 (d, 3H, H-16, J = 5.8 Hz) MS m/e: 585 [M.sup.+ + H] C3 14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy- cyclohexylsulfanyl]-acetyl}-12-desvinyl-12-[(3- amino-propylamino)-methyl] mutilin trihydrochlorid (comparator compound) MS m/e: 568 [M.sup.+ + H]
Antimicrobial Activity of Pleuromutilin Derivatives of Present Invention

(337) The antibacterial activity expressed as minimal inhibitory concentration (MIC) was determined according to the approved standard reference recommendations of CLSI (former NCCLS) as described in the description of the application.

(338) Compounds of the present invention exhibit very good activity against the clinical relevant bacterial pathogens Staphylococcus aureus and Escherichia coli. Surprisingly, the activity against Escherichia coli is significantly improved by introduction of C-12 substituents with inverted stereochemistry of the methyl group in position 12 of the mutilin ring (12-epi mutilins) according to the present invention compared with compounds wherein the stereochemistry of the methyl substituent in position 12 of the mutilin ring is as in the naturally occurring pleuromutilin. The in vitro activity against Escherichia coli of a compound of the present invention is significantly and surprisingly better than that of the comparator compound as set out in Table 22 below.

(339) TABLE-US-00022 TABLE 22 Antimicrobial activity of the compound of Example 1 and of the comparator compound C2 MIC [g/mL] ATCC Compound of Comparator Species number Example 1 Compound C2 Staphylococcus ATCC 49951 0.03 0.03 aureus Escherichia ATCC25922 0.5 32 coli

(340) Table 23 below presents the in vitro activity of further of compounds described in the present invention comprising surprising activity against Staphylococcus aureus and even more surprising activity against Escherichia coli.

(341) TABLE-US-00023 TABLE 23 Antimicrobial activity of the compounds of further examples described in the present invention MIC [g/mL] Compound S. aureus E. coli of Example ATCC 49951 ATCC25922 9 0.03 0.5 11 0.03 0.25 23 0.03 1 47 0.03 0.5 48 0.03 0.25 52 0.03 0.5 54 0.125 4 64 0.5 4 65 0.5 2 68 0.25 4 78 0.5 2 81 0.125 8 83 0.25 1 86 0.06 0.5 98 0.125 0.5 99 0.25 0.5 103 0.5 4 115 0.03 0.25 130 0.03 0.5 131 0.06 0.125 143 0.125 4 146 0.03 8 152 0.25 4 157 0.25 4

(342) Table 24 below presents the in vitro activity of

(343) the 12-epi-mutilin compound known from prior art (H. Berner et al, Monatshefte fir Chemie, 1986, 117, 1073-1080) (compound of Example C1) with the naturally occurring vinyl group at position C-12 and the methyl group in epi position, and

(344) the C-12-epi substituted compound of Example 66 with identical side chain at position C-14 of the mutilin ring as the compound of Example C1. Evidently the replacement of the naturally occurring vinyl group by a group according to the present invention in a compound having the methyl group in position C-12 of the mutilin ring inverted, namely contrary to the stereochemistry of the methyl group in position 12 in the natural occurring pleuromutilin ring, significantly and surprisingly improves the activity against Escherichia coli.

(345) TABLE-US-00024 TABLE 24 Antimicrobial activity of the compound of Example 66 and comparator of the compound of Example C1 MIC [g/mL] ATCC Compound of Species number Example 66 Compound C1 Staphylococcus ATCC 49951 0.5 0.5 aureus Escherichia ATCC25922 4 128 coli

(346) Table 25 below presents the in vitro activity of two compounds which are identical in structure with the only exception that the stereochemistry of the methyl substituent in position C-12 of the mutilin ring in a compound of the present invention is inverted whereas in the comparator compound the stereochemistry of said methyl group is the same as in the naturally occurring pleuromutilin. Thus, the C-12 epi compound of Example 62 exhibits significantly improved activity against Staphylococcus aureus and Escherichia coli compared to the comparator compound of Example C3 having the natural stereochemistry of the methyl group in position C-12 and otherwise is identical.

(347) TABLE-US-00025 TABLE 25 Antimicrobial activity of the compound of Example 62 and comparator compound C3 MIC [g/mL] ATCC Compound of Species number Example 62 Compound C3 Staphylococcus ATCC 49951 0.25 16 aureus Escherichia ATCC25922 4 >32 coli

(348) Table 26 below shows the activity of tiamulin, econor and retapamulin against Staphylococcus aureus and Escherichia coli. Whilst retaining good activity against Staphylococcus aureus the activity against Escherichia coli is significantly reduced with MIC >16 g/ml.

(349) TABLE-US-00026 TABLE 26 Antimicrobial activity of tiamulin, econor and retapamulin ATCC MIC [g/mL] Species number Tiamulin Econor Retapamulin Staphylococcus ATCC 49951 0.5 0.03 0.03 aureus Escherichia ATCC25922 >32 32 32 coli