THIENO-INDOLE MOIETIES AND METHODS OF TREATING USING THE SAME

Abstract

The present invention relates to a novel class of alkylating agents comprising a thieno-indole moiety linked to a DNA-binding moiety, which have cytotoxic activity and are useful in treating diseases such as cancer, cellular proliferation disorders and viral infections. The present invention also provides methods for preparing these compounds, pharmaceutical compositions comprising them and methods of treating diseases utilizing such compounds or the pharmaceutical composition containing them. The invention also relates to the use of this novel class of alkylating agents in the preparation of conjugates.

Claims

1.-21. (canceled)

22. A method for treating cancer, which comprises administering to a mammal in need thereof an effective amount of a compound of formula (II) ##STR00165## wherein R1 and R2 taken together form a group (D) or (G): ##STR00166## wherein R5 is hydrogen, linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.1-C.sub.4 hydroxyalkyl or linear or branched C.sub.1-C.sub.4 aminoalkyl; R3 and R4 are, each independently, hydrogen or a group selected from an optionally substituted linear or branched C.sub.1-C.sub.4 alkyl and linear or branched C.sub.1-C.sub.4 hydroxyalkyl; R6 is a leaving group; T is null or N; BM of formula (V): ##STR00167## wherein: X is null, linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.2-C.sub.4 alkenyl or linear or branched C.sub.2-C.sub.4 alkynyl; Y is an optionally substituted aryl or heteroaryl selected from the group consisting of ##STR00168## ##STR00169## Y is an optionally substituted aryl or heteroaryl selected from ##STR00170## ##STR00171## wherein R15, R16 and R20 are independently hydrogen, halogen, hydroxy, NO.sub.2, an optionally substituted linear or branched C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy, cyano, COOH, CONHR3, NC(O)OR3, C(NH)NH.sub.2 or NR3R4, and R3 and R4 are as defined above; U is a moiety of formula (VI) or (VII): ##STR00172## wherein R3 is as defined above; q is an integer from 0 to 4; L1 is hydrogen or L, wherein L is null or a moiety selected from NHCOR9 (Xa); NHCONHR9 (Xb); NHCOOR9 (Xc); NHR9 (Xd); ##STR00173## wherein R9 and R10 are, each independently, hydrogen, hydroxy or an optionally substituted group selected from linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.1-C.sub.4 hydroxyalkyl, linear or branched C.sub.1-C.sub.4 sulfhydrylalkyl and linear or branched C.sub.1-C.sub.4 aminoalkyl, when W1 and Z1 are null, or R9 and R10 are null when at least one of W1 and Z1 is not null, n is an integer from 0 to 2 and n1 is an integer from 0 to 4; W1 is null or a system comprising one or more groups independently selected from ##STR00174## ##STR00175## wherein one of R9 and R10 is null and the other is as defined above, R11 and R12 are, each independently, hydrogen, halogen, methyl, ethyl or linear or branched C.sub.1-C.sub.4 hydroxymethyl, m is an integer from 0 to 3, and A, is CH.sub.2, CH.sub.2NR12 or NR12, wherein R12 is as defined above; Z1 is null or a peptidic linker (Z.sub.a), a non peptidic linker (Z.sub.b) or hybrid linker (Z.sub.c), wherein Z.sub.a is selected from a single amino acid, a dipeptide, a tripeptide, a tetrapeptide and an oligopeptide moiety comprising natural L-amino acids, unnatural D-amino acids, synthetic amino acids or any combination thereof; Z.sub.b is selected from ##STR00176## wherein one of R9 and R10 is null and the other is as defined above and p is an integer from 1 to 20; and Z.sub.c is a group of formula
Z.sub.aZ.sub.b or Z.sub.bZ.sub.a wherein Z.sub.a and Z.sub.b are as defined above; provided that a compound of formula (II) wherein L1 is hydrogen is excluded when 1) both T and X are null, q is 0 and Y is an heterocyclyl moiety of formula (VIII), (VIII) or (VIII): ##STR00177## or 2) both T and X are null, q is 1, U is a group of formula (VII), Y is an heterocyclyl moiety of formula (IX) ##STR00178## and Y is an heterocyclyl moiety of formula (VIII) ##STR00179## wherein Q is NH or O, and R21 is hydrogen or a group selected from N(C.sub.2H.sub.5).sub.2 and C(NH)NH.sub.2; with the proviso that in a compound of formula (II) when L1 is L, at least one among L, W1 and Z1 is not null; or the pharmaceutically acceptable salts thereof, or a compound of formula (III) or (IV) ##STR00180## wherein R1 and R2 taken together form a group (D) or (G): ##STR00181## wherein R5 is hydrogen, linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.1-C.sub.4 hydroxyalkyl or linear or branched C.sub.1-C.sub.4 aminoalkyl; R3 and R4 are, each independently, hydrogen or a group selected from an optionally substituted linear or branched C.sub.1-C.sub.4 alkyl and linear or branched C.sub.1-C.sub.4 hydroxyalkyl; R6 is a leaving group; T is null or N; BM is of formula (V): ##STR00182## wherein: X is null, linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.2-C.sub.4 alkenyl or linear or branched C.sub.2-C.sub.4 alkynyl, Y is an optionally substituted aryl or heteroaryl selected from the group consisting of ##STR00183## ##STR00184## Y is an optionally substituted aryl or heteroaryl selected from ##STR00185## ##STR00186## wherein R15, R16 and R20 are independently hydrogen, halogen, hydroxy, NO.sub.2, an optionally substituted linear or branched C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy, cyano, COOH, CONHR3, NC(O)OR3, C(NH)NH.sub.2 or NR3R4, and R3 and R4 are as defined above, U is a moiety of formula (VI) or (VII) ##STR00187## wherein R3 is as defined above, and q is an integer from 0 to 4; A is an atom selected from O, NH, CO; A is null or A, wherein A is as defined above; L is null or a moiety selected from NHCOR9 (Xa); NHCONHR9 (Xb); NHCOOR9 (Xc); NHR9 (Xd); ##STR00188## wherein R9 and R10 are, each independently, hydrogen, hydroxy or an optionally substituted group selected from linear or branched C.sub.1-C.sub.4 alkyl, linear or branched C.sub.1-C.sub.4 hydroxyalkyl, linear or branched C.sub.1-C.sub.4 sulfhydrylalkyl and linear or branched C.sub.1-C.sub.4 aminoalkyl, when W, Z and RM are null, or R9 and R10 are null when at least one of W, Z and RM is not null, n is an integer from 0 to 2 and n1 is an integer from 0 to 4; L1 is hydrogen or L, wherein L is as defined above; W and W1 are independently null or a system comprising one or more groups independently selected from ##STR00189## ##STR00190## wherein one of R9 and R10 is null and the other is as defined above, R11 and R12 are, each independently, hydrogen, halogen, methyl, ethyl or linear or branched C.sub.1-C.sub.4 hydroxymethyl, m is an integer from 0 to 3, and A1 is CH.sub.2, CH.sub.2NR12 or NR12, wherein R12 is as defined above; Z and Z1 are independently null or a peptidic linker (Z.sub.a), a non peptidic linker (Z.sub.b) or a hybrid linker (Z.sub.c), wherein Z.sub.a is selected from a single amino acid, a dipeptide, a tripeptide, a tetrapeptide and an oligopeptide moiety comprising natural L-amino acids, unnatural D-amino acids, synthetic amino acids or any combination thereof; Z.sub.b is selected from ##STR00191## wherein one of R9 and R10 is null and the other is as defined above and p is an integer from 1 to 20; and Z.sub.c is a group of formula
Z.sub.aZ.sub.b or Z.sub.bZ.sub.a wherein Z.sub.a and Z.sub.b are as defined above; RM and RM1 are independently null or a moiety selected from ##STR00192## ##STR00193## wherein R13 is C.sub.1-C.sub.3 alkyl or an electron-withdrawing group selected from the group consisting of NO.sub.2 and CN; r is an integer from 0 to 7; and R11 and R12 are as defined above, said RM being attached to one or more of A, L, W or Z groups and said RM1 being attached to one or more of L1, W1 or Z1 groups; provided that 1) a compound of formula (IV) is excluded when A is null and RM1 is null; 2) a compound of formula (III) or (IV) is excluded when a) both T and X are null, q is 0 and Y is an heterocyclyl moiety of formula (VIII).sup.IV ##STR00194## wherein Q is O, S, NR14-, wherein R14 is hydrogen, C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.4 hydroxyalkyl; Q1 is CH or N; R7 and R8 are independently hydrogen, halogen, hydroxy, C.sub.1-C.sub.4 alkoxy, cyano, NCOOR3, C(NH)NH.sub.2 or NR3R4, wherein R3 and R4 are as defined above; or b) both T and X are null, q is 1 or 2, U is a group of formula (VII), Y is a heterocyclyl moiety of formula (IX): ##STR00195## Y is a heterocyclyl moiety of formula (VIII).sup.IV ##STR00196## wherein Q, Q1, R7 and R8 are as defined above; with the proviso that when L1 is L, at least one among L, W1, Z1 and RM1 is not null; and with the proviso that when A is A, at least one among L, W, Z and RM is not null; or the pharmaceutically acceptable salts thereof.

23. The method according to claim 22 wherein the mammal in need thereof is a human.

24. The method according to claim 22 wherein the cancer is selected from carcinoma of bladder, breast, colon, kidney, liver, lung, including small cell lung cancer, esophagus, gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, or skin, including squamous cell carcinoma; hematopoietic tumors of lymphoid lineage including leukaemia, acute lymphocytic leukaemia, acute lymphoblastic leukaemia, B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukaemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; tumors of the central and peripheral nervous system, including astrocytoma neuroblastoma, glioma and schwannomas; other tumors, including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoxanthoma, thyroid follicular cancer, Kaposi's sarcoma and mesothelioma.

25. The method according to claim 22, wherein the compound of formula (II), (III) or (IV), as defined in claim 1, is selected from the group consisting of: N-(6-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-3-yl)-1H-indole-6-carboxamide, N-(5-{[(8R)-8-(chloromethyl)-4-hydroxy-I-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1-methyl-1H-pyrrol-3-yl)-1-methyl-4-{[(1-methyl-4-nitro-1H-pyrrol-2-yl)carbonyl]amino}-1H-pyrrole-2-carboxamide, (2E)-1-[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-en-1-one, (2E)-1-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-en-1-one, (8R)N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-6-carboxamide, (8S)N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-6-carboxamide, N-(3-{(1E)-3-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-oxoprop-1-en-1-yl}-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide, N-(3-{(1E)-3-[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-oxoprop-1-en-1-yl}-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide, N-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indole-2-carboxamide, tert-butyl {2-[(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)carbamoyl]-1H-indol-5-yl}carbamate, (8S)-6-({5-[({5-[(tert-butoxycarbonyl)amino]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, (8S)-6-[(5-{[(5-amino-1H-indol-2-yl)carbonyl]amino)}-1H-indol-2-yl)carbonyl]-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate hydrochloride, (8S)-8-chloromethyl)-6-({5-[(1H-indol-2-ylcarbonyl)amino]-1H-indol-2-yl}carbonyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl piperazine-1-carboxylate, (8R)-6-({5-[({5-[(tert-butoxycarbonyl)amino]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, (8R)-6-[(5-1{[(5-amino-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate hydrochloride, (8S)-8-(chloromethyl)-1-methyl-6-[(5-{[(5-nitro-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, (8R)-8-(chloromethyl)-1-methyl-6-[(5-1{[(5-nitro-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, (8S)-8-(chloromethyl)-6-[(5-{[(5-hydroxy-1H-indol-2-yl)carbonyl]amino})-1H-indol-2-yl)carbonyl]-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, (8R)-8-(chloromethyl)-6-[(5-{[(5-hydroxy-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N5-carbamoyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N5-carbamoyl-N-[4 ({[{3-[({[(8R)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, [(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}methanone, [(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}methanone, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[({3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.6-carbamoyl-N-[4-({[{3-[[({(8R)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-omithinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-(({[(3-{[({(8S)-8-(chloromethyl)-1-methyl-6-[(2E)-3-{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}prop-2-enoyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl}oxy)carbonyl](methyl)amino}-2,2-dimethylpropyl)(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[(3-{([({(8R)-8-(chloromethyl)-1-methyl-6-[(2E)-3-{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}prop-2-enoyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl}oxy)carbonyl](methyl)amino}-2,2-dimethylpropyl)(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, (2E)-1-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-(1H-indol-3-yl)prop-2-en-1-one, N-(2-{[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1-methyl-1H-indol-5-yl)-1-methyl-1H-indole-2-carboxamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({-[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methy)carbamoyl]oxy}methy)phenyl]glycinamide, L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L -ornithinamide, L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl)}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L -valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl)}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide, N-[19-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-17-oxo-,7, 10,13-tetraoxa-16-azanonadecan-1-oyl]-L -valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-1-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-omithinamide and N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)methyl)amino]ethyl)}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide.

Description

EXAMPLE 1

Step b, Step a

N-(6-{[(8S)-8-{chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6]-yl}carbonyl}-1H-indol-3-yl)-1H-indole-6-carboxamide [(II)] (compd. 1)

[0238] ##STR00086##

Step b

[0239] A solution of (8S)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-ol (XVII) prepared as reported in the GB2344818 (14.2 mg, 0.0563 mmol) was dissolved in dry DMF (1.5 mL), and treated with EDCI (43 mg, 4 eq.) and 3-[(1H-indol-6-ylcarbonyl)amino]-1H-indole-6-carboxylic acid (XVI) (27 mg, 1.5 eq.) The mixture was stirred for 16 h at room temperature and then was quenched by adding saturated aqueous NaCl. Isolation of the product was performed by extraction with EtOAc (4) and subsequent washing of the combined organic layers with aqueous 2M HCl (3), saturated aqueous Na.sub.2CO.sub.3 (3) and saturated aqueous NaCl (3). Organic layer was dried (Na.sub.2SO.sub.4), concentrated under vacuum. The crude residue was purified by flash chromatography (hexane-acetone 6:4) to afford the title compound (18.7 mg, 60%).

[0240] ESI MS: m/z 555 (MH+)

##STR00087##

[0241] Analogously, by using the corresponding carboxylic acids, the following compounds have been prepared: N-(5-{[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1-methyl-1 H-pyrrol-3-yl)-1-methyl-4-{[(1-methyl-4-nitro-1H-pyrrol-2-yl)carbonyl]amino}1H-pyrrole-2-carboxamide [(II)](compd. 2)

##STR00088##

[0242] ESI MS: m/z 650 (MH+)

[0243] (2E)-1-[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[,2-e]indol-6-yl]-3-1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-en-1-one [(II)] (compd. 3)

##STR00089##

[0244] ESI MS: m/z 424 (MH+)

[0245] (2E)-1-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8 dihydro-6H-thieno[3,2-e]indol-6-yl]-3-1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-en-1-one [(II)] (compd. 4)

##STR00090##

[0246] ESI MS: m/z 424 (MH+)

[0247] N-(3-{(1E)-3-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2]indol-6-yl]-3 oxoprop-1-en-1-yl}1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide [(II)] (compd. 13)

##STR00091##

[0248] ESI MS: m/z 582 (MH+)

[0249] N-(3-{(1E) 3-[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]-3-oxoprop-1-en-1-yl}1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide [(II)] (compd. 14)

##STR00092##

[0250] ESI MS: m/z 582 (MH+)

[0251] (2E)-1-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6 yl]3-1H-indol-3-yl)prop-2-en-1-one [(II)] (compd. 43)

##STR00093##

[0252] ESI MS: m/z 423 (MH+)

[0253] .sup.1H NMR (500 MHz, acetone-d6) ppm 2.60 (d, J=1.0 Hz, 3H) 3.58 (m, 1H) 3.88 (m, J=11.2, 1.9 Hz, 1H) 4.2 (m, 1H) 4.46 (t, J=9.5 Hz, 1H) 4.63 (d, J=10.6 Hz, 1H) 7.07 (d, J=15.4 Hz, 1H) 7.24 (m, 2H) 7.32 (m, 1H) 7.53 (m, 1H) 7.91 (m, 1H) 8.02 (m, 2H) 8.13 (br. s., 1H) 9.26 (br. s., 1H) 10.81 (br. s., 1H)

[0254] N-(2-{[(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro 6H-thieno[3,2-e]indol-6 yl]carbonyl}-1-methyl-1H-indo-5-yl)-1-methyl-1H-indole-2-carboxamide [(II)] (compd. 44)

##STR00094##

[0255] ESI MS: m/z 583 (MH+)

Step a

[0256] N-(6-{[(3bR,4aS)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-3-yl)-1H-indole-6-carboxamide [(I)] (compd. 5)

[0257] Compound 1 (25 mg, 0.045 mmol) was dissolved in DMF (2 mL), and treated with a solution of NaHCO.sub.3 in water (1 mL, 15 mg NaHCO.sub.3/mL). The reaction mixture was stirred for 2 h, EtOAc was added and the resulting organic layer was washed with brine (4), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by flash chromatography (hexane-acetone 6:4) to afford compound 5 (19 mg, 82%).

##STR00095##

[0258] ESI MS: m/z 519 (MH+)

[0259] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.75 (t, J=4.8 Hz, 1H) 2.27 (s, 3H) 2.32 (dd, J=7.6, 4.6 Hz, 1H) 3.40 (dt, J=7.5, 4.9 Hz, 1H) 4.13 (d, J=11.3 Hz, 1H) 4.30 (dd, J=11.1, 4.7 Hz, 1H) 5.78 (s, 1H) 6.57 (br. s., 1H) 7.32 (dd, J=8.4, 0.7 Hz, 1H) 7.44 (s, 1H) 7.52 (t, J=2.7 Hz, 1H) 7.67 (d, J=8.2 Hz, 1H) 7.79 (m, 2H) 8.00 (d, J=8.2 Hz, 1H) 8.22 (d, J=2.7 Hz, 2H) 9.51 (s, 1H) 10.45 (br. s, 1H) 10.61 (br. s, 1H)

[0260] Analogously, by using the corresponding carboxylic acids, the following compounds have been prepared:

[0261] 1-methyl-N-(1-methyl-5-{[(3bS,4aR)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-pyrrol-3-yl)-4-4{[(1-methyl-4-nitro-1H-pyrrol-2-yl)carbonyl]amino}-1H-pyrrole-2-carboxamide [(I)] (compd. 6)

##STR00096##

[0262] ESI MS: m/z 614 (MH+)

[0263] .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 1.52 (t, J=4.9 Hz, 1H) 2.21 (s, 3H) 2.28 (dd, J=7.6, 4.3 Hz, 1H) 3.43 (dt, J=7.8, 5.2 Hz, 1H) 3.76 (s, 3H) 3.86 (s, 3H) 3.96 (s, 3H) 4.16 (d, J=10.7 Hz, 1H) 4.29 (dd, J=10.6, 4.8 Hz, 1H) 6.30 (s, 1H) 6.77 (d, J=1.7 Hz, 1H) 7.08 (d, J=1.8 Hz, 1H) 7.25 (d, J=1.7 Hz, 1H) 7.48 (d, J=1.7 Hz, 1H) 7.58 (t, J=1.9 Hz, 2H) 8.19 (d, J=1.8 Hz, 1H) 10.00 (s, 1H) 10.30 (s, 1H)

[0264] (3bS,4aR)-3-methyl-6-[(2E)-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-enoyl]-4,4a,5,6-tetrahydro-8H-cyclopropa[c]thieno[3,2-e]indo-8-one [(I)] (compd. 7)

##STR00097##

[0265] ESI MS: m/z 388 (MH+)

[0266] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.45 (t, J=4.8 Hz, 1H) 2.19 (dd, J=7.6, 4.5 Hz, 1H) 2.26 (d, J=0.7 Hz, 3H) 3.44 (dt, J=7.7, 4.8 Hz, 1H) 4.40 (dd, J=10.4, 4.8 Hz, 1H) 4.51 (d, J=10.3 Hz, 1H) 6.98 (d, J=15.4 Hz, 1H) 7.08 (br. s., 1H) 7.24 (dd, J=7.9, 4.7 Hz, 1H) 7.44 (s, 1H) 7.98 (d, J=15.4 Hz, 1H) 8.09 (s, 1H) 8.34 (dd, J=4.8, 1.5 Hz, 1H) 8.41 (dd, J=7.8, 1.3 Hz, 1H) 11.30 (s, 1H)

[0267] (3bR,4aS)-3-methyl-6-[(2E)-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)prop-2-enoyl]-4,4a,5,6-tetrahydro-8H-cyclopropa[c]thieno[3,2-e]indol-8-one [(I)] (compd. 8)

##STR00098##

[0268] ESI MS: m/z 388 (MH+)

[0269] .sup.1H NMR (500 MHz, acetone-d6) ppm 1.45 (1, J=4.9 Hz, 1H) 2.16-2.21 (m, 1H) 2.26 (s, 3H) 3.44 (dt, J=7.7, 4.9 Hz, 1H) 4.40 (dd, J=10.4, 5.1 Hz, 1H) 4.51 (d, J=10.4 Hz, 1H) 6.98 (d, J=15.4 Hz, 1H) 7.08 (br. s., 1H) 7.24 (dd, J=7.8, 4.5 Hz, 1H) 7.44 (s, 1H) 7.98 (d, J=15.4 Hz, 1H) 8.09 (s, 1H) 8.34 (dd, J=4.7, 1.1 Hz, 1H) 8.41 (d, J=7.8 Hz, 1H) 11.29 (br. s., 1H)

[0270] N-(3-{(1E)-3-[(3bR,4aS)-3-methyl-8-oxo 4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]3-oxoprop-1-en-1-yl}-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide [(I)] (compd. 15)

##STR00099##

[0271] ESI MS: m/z 546 (MH+)

[0272] N-(3-{(1E)-3-[(3bR,4aS)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]-3-oxoprop-1-en-1-yl}-1H-pyrrolo[2,3-b]pyridin-5-yl)-1H-indole-2-carboxamide [(I)] (compd. 16)

##STR00100##

[0273] ESI MS: m/z 546 (MH+)

[0274] .sup.1H NMR (500 MHz, acetone-d6) ppm 1.58 (m, 1H) 2.24 (m, 1H) 2.27 (s, 3H) 3.47 (dt, J=7.6, 4.9 Hz, 1H) 4.39 (dd, J=10.2, 4.9 Hz, 1H) 4.48 (m, 1H) 6.96 (d, J=15.1 Hz, 1H) 7.01 (s, 1H) 7.12 (m, 2H) 7.28 (m, 1H) 7.40 (d, J=1.8 Hz, 1H) 7.45 (s, 1H) 7.61 (d, J=8.3 Hz, 1H) 7.70 (d, J=8.1 Hz, 1H) 7.98 (m, 1H) 8.09 (d, J=2.8 Hz, 1H) 8.76 (d, J=2.0 Hz, 1H) 8.87 (d, J=2.0 Hz, 1H) 9.76 (br. s., 1H) 10.97 (br. s., 1H) 11.32 (br. s., 1H)

[0275] 1-methyl-N-(1-methyl-2-{[(3bS,4aR)-3-methy-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)-1H-indole-2-carboxamide [(I)] (compd. 45)

##STR00101##

[0276] ESI MS: m/z 547 (MH+)

[0277] .sup.1H NMR (500 MHz, acetone-d6) ppm 1.65 (t, J=4.9 Hz, 1H) 2.27 (d, J=12.4 Hz, 1H) 2.27 (d, J=0.7 Hz, 3H) 3.43 (dt, J=7.6, 4.89 Hz, 1H) 3.93 (s, 3H) 4.12 (s, 3H) 4.38 (d, J=11.1 Hz, 1H) 4.51 (dd, J=10.8, 4.8 Hz, 1H) 6.52 (s, 1H) 7.08 (s, 1H) 7.13 (m, 1H) 7.26 (s, 1H) 7.32 (td, J=7.7, 1.0 Hz, 1H) 7.47 (d, J=1.0 Hz, 1H) 7.54 (m, 2H) 7.66 (d, J=7.8 Hz, 1H) 7.71 (dd, J=9.1, 2.0 Hz, 1H) 8.29 (d, J=2.0 Hz, 1H) 9.54 (s, 1H)

EXAMPLE 2

tert-butyl {2-[(2-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1 methyl-7,8-dihydro-6H-thieno[32-e]indol-6-yl]carbonyl}-1 H-indol-5-yl)carbamoyl]-1H-indol-5-yl)carbamate [(II)] (compd. 18)

[0278] Step b, deprotection, step a

##STR00102##

Step b

[0279] A solution of (8S)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-ol ((XVII), 11.4 mg, 0.045 mmol), prepared as reported in GB2344818, was dissolved in dry DMF (1 mL), and treated with EDCI (35 mg, 4 eq.) and 5-[({5-[(tert-butoxycarbonyl)amino]-1H-indol-2-yl}carbonyl)amino]-1H-indole-2-carboxylic acid (XVI) (29 mg, 1.5 eq.) The mixture was stirred for 16 h at room temperature and then was quenched by adding saturated aqueous NaCl. Isolation of the product was performed by extraction with EtOAc (4) and subsequent washing of the combined organic layers with aqueous 2M HCl (3), saturated aqueous Na.sub.2CO.sub.3 (3) and saturated aqueous NaCl (3). Organic layer was dried (Na.sub.2SO.sub.4), concentrated under vacuum to give the title compd. 18, that is then purified by flash chromatography hexane-acetone 1:1).

##STR00103##

[0280] ESI MS: m/z 670 (MH.sup.+)

[0281] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.50 (s, 9H) 1.79 (dt, J=6.4, 3.3 Hz, 1H) 2.61 (s, 2H) 3.53-3.66 (m, 1H) 3.89 (dd, J=11.4, 2.8 Hz, 1H) 4.25 (m, 1H) 4.73 (m, 1H) 4.84 (d, J=10.6 Hz, 1H) 7.21 (s, 1H) 7.26 (s, 1H) 7.34 (m, 2H) 7.49 (d, J=8.8 Hz, 1H) 7.56 (m, 1H) 7.61 (m, 1H) 7.96 (br. s., 2H) 8.21 (br. s., 1H) 8.34 (s, 1H) 9.28 (s, 1H) 9.52 (s, 1H) 10.73 (br. s., 1H) 10.80 (br. s., 1H).

[0282] By deprotection, the following compound has been obtained: 5-amino-N-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-5a,7,8,8a-tetrahydro-6H-thieno[3,2-e]indo-6-yl]carbonyl}-1H-indol-5-yl)-1H-indole-2-carboxamide [(II)] (compd. 19)

##STR00104##

[0283] A solution of compd. 18 (18 mg, 0.0268 mmol) in 3.5 M HCl-EtOAc (5 mL) was stirred for 30 minutes before removing the solvent under a steady stream of nitrogen and affording hydrochloride derivative of compd. 2 (15 mg, 89%).

[0284] ESI MS: m/z 570 (MH.sup.+)

Step a

[0285] tert-butyl {2-[(2-{[(3bR,4aS)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)carbamoyl]-1H-indol-5-yl}carbamate [(I)] (compd. 20)

##STR00105##

[0286] Compd. 18 (30 mg, 0.045 mmol) was dissolved in DMF (1 mL), and treated with aqueous NaHCO.sub.3 (0.5 mL, 15 mg NaHCO.sub.3/mL). The reaction mixture was stirred for 2 h, EtOAc was added and the resulting organic layer was washed with brine (4), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by flash chromatography (hexane-acetone 6:4) to afford the final compd. 20 (9 mg, 31%).

[0287] ESI MS: m/z 634 (MH.sup.+)

[0288] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.41 (s, 9H) 1.56 (t, J=4.8 Hz, 1H) 2.22 (m, 1H) 2.29 (d, J=0.9 Hz, 3H) 3.52 (m, 1H) 4.69 (m, 2H) 7.06 (s, 1H) 7.26 (d, J=1.5 Hz, 1H) 7.28 (d, J=1.8 Hz, 1H) 7.36 (dd, J=8.8, 1.8 Hz, 1H) 7.48 (m, 1H) 7.49 (d, J=8.8 Hz, 1H) 7.58 (m, 1H) 7.63 (m, 1H) 7.96 (br. s., 1H) 8.26 (br. s., 1H) 8.35 (s, 1H) 9.55 (s, 1H) 10.77 (br. s., 1H) 10.95 (br. s., 1H)

[0289] By deprotection the following compound has been prepared:

[0290] 5-amino-N-(2-{[(3bR,4aS)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)-1H-indole-2-carboxamide [(I)] (compd. 21)

##STR00106##

[0291] ESI MS: m/z 534 (MH.sup.+)

[0292] By analogue procedure and using the suitable starting material the following compounds have been prepared:

[0293] tert-butyl {2-[(2-{[(3bS,4aR)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)carbamoyl]-1H-indol-5-yl}carbamate [(I)] (compd. 25)

##STR00107##

[0294] ESI MS: m/z 634 (MH.sup.+)

[0295] 5-amino-N-(2-{[(3bS,4aR)-3-methy oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)-1H-indole-2-carboxamide [(1)] (compd. 26)

##STR00108##

[0296] ESI MS: m/z 534 (MH.sup.+)

[0297] N-(2-{[(3bR,4aS)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)-5-nitro-1H-indole-2-carboxamide [(I)] (compd. 29)

##STR00109##

[0298] ESI MS: m/z 564 (MH.sup.+)

[0299] N-(2-{[(3bS,4aR)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indol-6(8H)-yl]carbonyl}-1H-indol-5-yl)-5-nitro-1H-indole-2-carboxamide [(I)] (compd. 30)

##STR00110##

[0300] ESI MS: m/z 564 (MH.sup.+)

EXAMPLE 3

Step b, Deprotection, Step a

[0301] ##STR00111##

Step b

The Intermediate (8R)N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-4-(benzyloxy)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-carboxamide

[0302] A solution of 4-amino-N-[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]-1-methyl-1H-pyrrole-2-carboxamide, prepared as reported in J.Med. Chem. 2004, 47, 2611-2623, (155 mg, 0.341 mmol), triethylamine (0.048 mL, 0.341 mmol) and CDI (300 mg, 1.71 mmol) in dry DMF (5 mL) was stirred at room temperature for 1 h. After evaporation of the solvent, the residue was treated with THF and stirred overnight. The precipitate (comp XVI) was filtered and dried under vacuum at 50 C. and treated with (8R)-4-(benzyloxy)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole (comp (XVII) (15.5 mg, 0.045 mmol, prepared as reported in the GB2344818) in dry DMF (2 mL) in the presence of NaHCO.sub.3 (4 mg, 0.047 mmol) and were stirred under nitrogen atmosphere at room temperature for 16 h. After evaporation of the solvent the residue was purified by flash chromatography (DCM-MeOH 95:5) to afford the intermediate (30 mg, 80%).

##STR00112##

[0303] ESI MS: m/z 824 (MH+)

[0304] .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 2.31 (t, J=7.2 Hz, 2H) 2.53 (s, 3H) 3.53 (t, J=10.4 Hz, 1H) 3.80 (s, 3H) 3.85 (s, 3H) 3.87 (s, 3H) 4.12 (m, 1H) 4.18 (m, 1H) 4.32 (d, J=10.4 Hz, 1H) 5.27 (s, 2H) 6.83 (d, J=1.7 Hz, 1H) 6.84 (m, 1H) 7.05 (m, 2H) 7.13 (d, J=1.5 Hz, 1H) 7.20 (d, J=1.5 Hz, 1H) 7.25 (d, J=1.5 Hz, 1H) 7.36 (m, 2H) 7.41 (s, 1H) 7.43 (m, 2H) 7.50 (d, J=7.3 Hz, 2H) 7.99 (m, 2H) 8.77 (s, 1H) 9.91 (s, 1H) 9.93 (s, 1H)

[0305] Analogously, by using the corresponding acyl derivative, the following compounds have been prepared: (8S)N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl)carbamoyl}-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-yl)-4-(benzyloxy)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-6-carboxamide

##STR00113##

[0306] ESI MS: m/z 824 (MH+)

[0307] N-(2-{[(8S)-4-(benzyloxy)-8-(chloromethyl)-1-methy-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indole-2-carboxamide

##STR00114##

[0308] ESI MS: m/z 758 (MH+)

[0309] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.74 (m, 4H) 2.59 (m, 4H) 2.63 (d, J=1.0 Hz, 3H) 2.86 (m, 2H) 3.65 (dd, J=11.4, 9.6 Hz, 1H) 3.93 (dd, J=11.4, 2.4 Hz, 1H) 4.12 (t, J=6.1 Hz, 2H) 4.29 (m, 1H) 4.77 (m, 1H) 4.87 (m, 1H) 5.36 (m, 2H) 6.94 (dd, J=8.8, 2.3 Hz, 1H) 7.15 (d, J=2.3 Hz, 1H) 7.25 (m, 2H) 7.35 (m, 1H) 7.38 (d, J=1.0 Hz, 1H) 7.43 (m, 2H) 7.49 (d, J=9.1 Hz, 1H) 7.58 (m, 3H) 7.63 (m, 1H) 8.18 (br. s., 1H) 8.37 (d, J=1.3 Hz, 1H) 9.52 (s, 1H) 10.77 (br. s., 1H) 10.87 (br. s., 1H)

[0310] [(8S)-4-(benzyloxy)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}methanone hydrochloride

##STR00115##

[0311] ESI MS: m/z 600 (MH+)

[0312] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 2.06 (m, 4H) 2.62 (d, J=1.0 Hz, 3H) 3.02-3.32 (m, 2H) 3.56 (br. s., 2H) 3.63 (dd, J=11.4, 9.6 Hz, 1H) 3.91 (dd, J=11.4, 2.4 Hz, 1H) 4.27 (m, 1H) 4.57 (br. s., 2H) 4.73 (dd, J=10.2, 8.2 Hz, 1H) 4.84 (m, 1H) 5.35 (m, 2H) 7.05 (dd, J=8.8, 2.3 Hz, 1H) 7.16 (d, J=1.7 Hz, 1H) 7.31 (d, J=1.8 Hz, 1H) 7.35 (m, 1H) 7.38 (d, J=1.0 Hz, 1H) 7.43 (m, 2H) 7.52 (d, J=9.1 Hz, 1H) 7.57 (d, J=7.6 Hz, 2H) 8.16 (br. s., 1H) 10.80 (br. s., 1H) 13.38 (br. s., 1H)

Deprotection

[0313] (8R)N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-6-carboxamide [(II)] (compd. 10)

[0314] A solution of the intermediate (15 mg, 0.0182 mmol), 25% aq. HCO.sub.2NH.sub.4 (0.15 mL) and 10% PdC (15 mg) in THF (3 mL) was stirred for 3 h under nitrogen atmosphere. The reaction mixture was filtered through Celite and concentrated to yield after chromatographic purification (DCM-MeOH 10:1) compound 10 (6.7 mg, 50%).

##STR00116##

[0315] ESI MS: m/z 734 (MH+)

[0316] Analogously the following compounds have been prepared:

[0317] (8S)N-(5-{[5-{5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indole-6-carboxamide [(II)] (compd. 11)

##STR00117##

[0318] ESI MS: m/z 734 (MH+)

[0319] N-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indole-2-carboxamide [(II)] (compd. 17)

##STR00118##

[0320] ESI MS: m/z 668 (MH+)

[0321] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 2.19-2.25 (m, 4H) 2.60 (s, 3H) 3.39 (br. s., 2H) 3.81 (m, 2H) 3.88 (dd, J=11.1, 2.5 Hz, 1H) 3.91-3.96 (br. s, 2H) 4.24 (m, 1H) 4.51 (m, 1H) 4.72 (dd, J=10.8, 7.8 Hz, 1H) 4.80-4.82 (d, J=10.8, 1 H) 7.01 (dd, J=8.9, 2.4 Hz, 1H) 7.22 (s, 1H) 7.26 (d, J=2.3 Hz, 1H) 7.28 (s, 1H) 7.33 (s, 1H) 7.49 (d, J=8.9 Hz, 1H) 7.55 (m, 1H) 7.60 (m, 1H) 7.90 (br. s., 1H) 8.29 (d, J=1.8 Hz, 1H)

[0322] [(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}methanone hydrochloride [(II)] (compd. 37)

##STR00119##

[0323] ESI MS: m/z 510 (MH+)

[0324] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 2.10-2.29 (m, 4H) 2.58 (d, J=1.3 Hz, 3H) 3.33 (m, 2H) 3.54 (dd, J=11.4, 10.1 Hz, 1H) 3.80 (m, 2H) 3.89 (m, 3H) 4.20 (m, 1H) 4.51 (m, 2H) 4.68 (dd, J=10.2, 8.2 Hz, 1H) 4.77 (m, 1H) 7.04 (dd, J=8.9, 2.4 Hz, 1H) 7.12 (s, 1H) 7.29 (d, J=2.3 Hz, 1H) 7.31 (d, J=0.5 Hz, 1H) 7.49 (d, J=8.9 Hz, 1H) 7.87 (br. s., 1H)

[0325] [(8R)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]{5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl})methanone hydrochloride [(II)] (compd. 38)

##STR00120##

[0326] ESI MS: m/z 510 (MH+)

Step a

[0327] A solution of compound 10 (5 mg, 0.0068 mmol), in DMF (5 mL), was treated with a solution of NaHCO.sub.3 in water (3 mL, 15 mg NaHCO.sub.3/mL). The reaction mixture was stirred for 4 h, EtOAc was added and the resulting organic layer was washed with brine (4), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by flash chromatography (DCM-MeOH 10:1) to afford the compound 9 (3.9 mg, 82%).

[0328] (3bS,4aR)-N-(5-{[5-({5-[(3-amino-3-oxopropyl)carbamoyl]-1-methyl-1H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indole-6(8H)-carboxamide [(I)] (compd. 9)

##STR00121##

[0329] ESI MS: m/z 698 (MH+)

[0330] .sup.1H NMR (500 MHz, dichloromethane-d.sub.2) ppm 1.37 (m, 1H) 2.12 (dd, J=7.6, 4.6 Hz, 1H) 2.18 (d, J=0.9 Hz, 3H) 2.47 (t, J=6.6 Hz, 2H) 3.26 (dt, J=7.6, 5.0 Hz, 1H) 3.53 (1, J=6.6 Hz, 2H) 3.85 (s, 3H) 3.88 (s, 6H) 4.05 (d, J=10.4 Hz, 1H) 4.14 (m, 1H) 6.64 (d, J=2.1 Hz, 1H) 6.73 (s, 1H) 6.82 (m, 2H) 6.99 (d, J=1.8 Hz, 1H) 7.17 (d, J=1.8 Hz, 1H) 7.20 (d, J=1.8 Hz, 1H) 7.29 (d, J=0.9 Hz, 1H)

[0331] Analogously the following compounds have been prepared:

[0332] (3bR,4aS)N-(5-{5-({5-[(3-amino-3-oxopropyl)carbamoyl-1-methyl-H-pyrrol-3-yl}carbamoyl)-1-methyl-1H-pyrrol-3-yl]carbamoyl}-1-methyl-1H-pyrrol-3-yl)-3-methyl-8-oxo-4a,5-dihydro-4H-cyclopropa[c]thieno[3,2-e]indole-6(8H)carboxamide [(I)] (compd. 12)

##STR00122##

[0333] ESI MS: m/z 698 (MH+)

EXAMPLE 4

Step c, Deprotection

[0334] ##STR00123##

Step c

[0335] (8S)-6-({5-[({5-[(tert-butoxycarbonyl)amino]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate [(II)](compd. 22)

##STR00124##

[0336] To a solution of tert-butyl {2[(2-[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)carbamoyl]-1H-indol-5-yl)carbamate, (compd. 18) (42 mg, 0.064 mmol) in dry DCM (6 mL) 4-methylpiperazine-1-carbonyl chloride hydrochloride (XIII) (39 mg, 0.193 mmol) and N,N-dimethylaminopyridine (27 mg, 0.212 mmol) were added. The reaction mixture was stirred at room temperature under nitrogen atmosphere for 16 h. The solvent was evaporated and the residue was dissolved in EtOAc, the resulting organic layer was washed with brine (3), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by flash chromatography (DCM-MeOH 95:5) to afford the compd. 22 (30 mg, 59%).

[0337] ESI MS: m/z 796 (MH.sup.+)

[0338] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 2.22 (s, 3H) 2.65 (d, J=0.9 Hz, 3H) 2.39-2.91 (m, 4H) 3.72 (dd, J=11.6, 9.5 Hz, 1H) 3.96 (dd, J=11.6, 3.1 Hz, 1H) 3.49-4.14 (m, 4H) 4.37 (m, 1H) 4.79-4.85 (m, 1H) 4.87-4.93 (m, 1H) 7.27 (d, J=1.5 Hz, 1H) 7.28 (s, 1H) 7.34-7.38 (m, 1H) 7.41 (s, 1H) 7.50 (d, J=8.8 Hz, 1H) 7.56 (d, J=8.5 Hz, 1H) 7.62-7.65 (m, 1H) 7.96 (br. s., 1H) 8.38 (s, 1H) 9.60 (s, 1H) 10.86 (br. s., 1H) 10.90 (br. s., 1H)

[0339] By analogous procedure the following products have been prepared:

[0340] (8R)-6-[5-{[(5-[(tert-butoxycarbonyl)amino]-1H-indol-2-yl)carbonyl]amino}-1H-indo-2-yl)carbonyl]-8-(chloromethyl)-1-methy-7,8-dihydro-6H-thieno[3,2-e]indo-4-yl 4-methylpiperazine-1-carboxylate [(II)](compd. 27) p

##STR00125##

[0341] ESI MS: m/z 796 (MH.sup.+)

[0342] (8S)-8-(chloromethyl)-1-methy-6-[(5-{[(5-nitro-1H-idol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl-4-methylpiperazine-1-piperazine-1-carboxylate [(II)] (compd. 31)

##STR00126##

[0343] ESI MS: m/z 726 (MH.sup.+)

[0344] (8R) 8-(chloromethyl)-1-methyl-6-[(5-{[(5-nitro-1H-indol-2-yl)carbonyl]amino}-1H-indo-2-yl)carbonyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate [(II)] (compd. 32)

##STR00127##

[0345] ESI MS: m/z 726 (MH.sup.+)

Deprotection

[0346] (8S)-6-[(5-{[(5-amino-1H-indol-2-yl)carbonyl]amino}-1H-indo-2-yl)carbonyl]-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate hydrochloride [(II)] (compd. 23)

##STR00128##

[0347] A solution of compd. 22 (22 mg, 0.0276 mmol) in 3.5 M HCl-EtOAc (5 mL) was stirred for 30 minutes before removing the solvent under a steady stream of nitrogen and affording the desired product as hydrochloride salt (18 mg, 89%).

[0348] ESI MS: m/z 696 (MH.sup.+)

[0349] .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 2.59 (s, 3H) 2.76 (br. s, 3H) 3.05-3.42 (m, 4H) 3.75 (dd, J=10.8, 7.9 Hz, 1H) 3.95-4.01 (m, 1H) 3.58-4.08 (m, 4H) 4.32-4.40 (m, 1H) 4.67 (d, J=11.1 Hz, 1H) 4.75-4.83 (m, 1H) 6.92 (br. s., 1H) 7.21 (br. s., 1H) 7.23 (s, 1H) 7.30 (br. s., 1H) 7.37 (d, J=8.4 Hz, 1H) 7.49-7.52 (m, 1H) 7.55 (s, 1H) 7.57-7.60 (m, 1H) 8.16 (s, 1H) 8.24 (s, 1H) 10.15 (br. s., 1H) 11.67 (br. s., 1H) 11.70 (s, 1H)

[0350] By analogous procedure the following products have been prepared:

[0351] (8R)-6-[(5-{[(5-amino-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-8-(chloromethyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate hydrochloride [(II)] (compd. 28)

##STR00129##

[0352] ESI MS: m/z 697 (MH.sup.+)

[0353] (8S)-8-(chloromethyl)-6-[(5-{[(5-hydroxy-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-1-ethyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate [II)] (compd. 33)

##STR00130##

[0354] ESI MS: m/z 697 (MH.sup.+)

[0355] (8R)-8-(chloromethyl)-6-[(5-{[(5-hydroxy-1H-indol-2-yl)carbonyl]amino}-1H-indol-2-yl)carbonyl]-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl 4-methylpiperazine-1-carboxylate [(II)] (compd. 34)

##STR00131##

[0356] ESI MS: m/z 697 (MH.sup.+)

EXAMPLE 5

Step c, Deprotection

[0357] ##STR00132##

Step c

[0358] Preparation of tert-butyl (8S)-8-(chloromethyl)-6-({5-[(1H-indol-2-ylcarbonyl)amino]-1H-indol-2-yl}carbonyl) -1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl piperazine-1,4-dicarboxylate [(II)]

##STR00133##

[0359] N-(2-{[(8S)-8-chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-1H-indole-2-carboxamide (111 mg, 0.2 mmol), prepared as reported in GB2344818, was dissolved in dry DCM (15 mL) and to this solution tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (100 mg, 0.4 mmol) and N,N-dimethylaminopyridine (55 mg, 0.45 mmol) were added. The reaction mixture was stirred at room temperature under nitrogen atmosphere for 16 h. The solvent was evaporated and the residue was dissolved in EtOAc, the resulting organic layer was washed with brine (4), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by flash chromatography (hexane-acetone 7:3) to afford the title compound (30 mg, 19%).

[0360] ESI MS: m/z 767 (MH.sup.+)

[0361] .sup.1H NMR (500 MHz, acetone-d.sub.6) ppm 1.48 (s, 9H) 2.65 (d, J=1.01 Hz, 3H) 3.55 (br. s., 4H) 3.60 (br. s., 2H) 3.73 (dd. J=11.36, 9.59 Hz, 1H) 3.79 (br. s., 2H) 3.97 (dd, J=11.36, 2.78 Hz, 1H) 4.33-4.42 (m, 1H) 4.80-4.86 (m, 1H) 4.89-4.94 (m, 1H) 7.10 (t, J=7.19 Hz, 1H) 7.23-7.27 (m, 1H) 7.28 (s, 1H) 7.34 (d, J=1.51 Hz, 1H) 7.42 (s, 1H) 7.53-7.57 (m, 1H) 7.60 (d, J=8.08 Hz, 1H) 7.61-7.65 (m, 1H) 7.67 (d, J=8.08 Hz, 1H) 8.29 (s, 1H) 8.38 (d, J=1.26 Hz, 1H) 9.56 (s, 1H) 10.89 (br. s., 2H).

Deprotection

[0362] (8S)-8-(chloromethyl)-6-({5-[(1H-indo-2-yl}carbonyl)amino]-1H-indo-2-ylcarbonyl)-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl piperazine-1-carboxylate [(II)] (compd. 24)

##STR00134##

[0363] A solution of the intermediate (25 mg, 0.0326 mmol) in 3.5 M HCl-EtOAc (5 mL) was stirred for 2 h. After evaporation of the solvent under a steady stream of nitrogen, the residue was dried in vacuo to afford the compd. 9 (11 mg, 48%).

[0364] ESI MS: m/z 667 (MH.sup.+)

EXAMPLE 6

Step e

[0365] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N5carbamoyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (compd. 35)

##STR00135##

Step e

[0366] To a solution of N-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indole-2-carboxamide (Compd 17) (5 mg, 0.0071 mmol) in DCM (1 ml) and THF (0.5 mL) were added 4-nitrophenyl chloroformate (3.1 mg, 0.0156 mmol) and triethylamine (2.2 mL, 0.0156 mmol). The resulting mixture was stirred at room temperature for 6 hours. N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-{4-[({[2,2-dimethyl-3-(methylamino)propyl] (methyl)carbamoyl}oxy)methyl]phenyl}-L-omithinamide (XX) (16 mg, 0.018 mmol) and triethylamine (3 mL, 0.021 mmol) were added. The mixture thus obtained was stirred at room temperature overnight, diluted with DCM and washed with a saturated solution of aqueous NaHCO.sub.3. The crude residue was purified by flash chromatography (DCM-MeOH 100:15) to afford the title compound (2 mg, 20%).

[0367] ESI MS: m/z 1422 (MH+)

[0368] .sup.1H NMR (500 MHz, methanol-d.sub.3) ppm 0.82-1.45 (m 29H) 2.68 (s, 3H) 2.90-3.61 (m, 20H) 3.84-4.34 (m, 8H) 4.49 (m, 1H) 5.04-5.14 (m, 2H) 6.70-6.77 (m, 2H) 7.01 (dd, J=9.0, 2.4, 1H) 7.18-7.71 (m, 11H) 8.11 (br. s., 2H)

[0369] Analogously the following compounds have been prepared:

[0370] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1 yl)hexanoyl]-L-valyl-N5-carbamoyl-N[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-omithinamide [(IV)] (Compd 36).

##STR00136##

[0371] ESI MS: m/z 1424 (MH.sup.+)

[0372] N-[6-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd. 39)

##STR00137##

[0373] N-[6-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{3-[({[(8R)-8-(chloromethyl)-1-methyl-6-({5-[2-pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd 40).

##STR00138##

[0374] ESI MS: m/z 1264 (MH+)

[0375] N-[6-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[(3-{[({(8S)-8-(chloromethyl)-1-methyl-6-[(2E)-3-{5-[2-pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}prop-2-enoyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl}oxy)carbonyl](methyl)amino}-2,2-dimethylpropyl)(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd 41).

##STR00139##

[0376] ESI MS: m/z 1290 (MH+)

[0377] N-[6-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[(3-{[({(8R)-8-(chloromethyl)-1-methyl-6-[(2E)-3-{-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}prop-2-enoyl]-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl}oxy)carbonyl](methy)amino}-2,2-dimethylpropyl)(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd. 42)

##STR00140##

[0378] ESI MS: m/z 1290 (MH+)

[0379] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide [(IV)] (Compd 46)

##STR00141##

[0380] ESI MS: m/z 1483 (MH+)

[0381] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{3-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]-2,2-dimethylpropyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide [(IV)] (Compd 47)

##STR00142##

[0382] ESI MS: m/1325 (MH+)

[0383] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide [(IV)] (Compd 48)

##STR00143##

[0384] ESI MS: m/z 1441 (MH+)

[0385] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1 yl)hexanoyl]-L-phenylalanyl-L-leucyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methy-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indo-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]glycinamide [(IV)](Compd 49)

##STR00144##

[0386] ESI MS: m/z 1283 (MH+)

EXAMPLE 7

Step c, Deprotection, Step g

[0387] ##STR00145## ##STR00146##

Step c

N-[(9H fluoren-9-ylmethoxy)carbonyl]-L-valyl-N.SUP.5.-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2 yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(II)]

[0388] ##STR00147##

[0389] To a solution of N-(2-{[(8S)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-6H-thieno[3,2-e]indol-6-yl]carbonyl}-1H-indol-5-yl)-5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indole-2-carboxamide hydrochloride (13 mg, 0.02 mmol) in THF/DCM 1:1 (2 mL) at 0 C. under nitrogen atmosphere, were added 4-nitrophenyl chloroformate (20 mg, 0.1 mmol) and triethylamine (20 mL, 0.14 mmol). The mixture was stirred at room temperature for 6 hours, solvent was evaporated and the residue was isolated by filtration after treatment with diethyether. The resulting solid compound was diluted with 1 mL of DCM (10% DMF) and N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-valyl-N.sup.5-carbamoyl-N-{4-[({methyl[2-(methylamino)ethyl]carbamoyl}oxy)methyl]phenyl}-L-omithinamide hydrochloride (38 mg, 0.05 mmol) and trietylamine (7 mL 0.05 mmol) were added. The mixture was stirred overnight, solvents were evaporated and title compound (12 mg, 42% yield) was isolated by column chromatography purification (DCM/MeOH 8:2).

[0390] ESI MS: m/z 1409 (MH+)

[0391] .sup.1H NMR (500 MHz, methanol-d.sub.4) ppm 2.07 (br. s., 4H) 2.58 (s, 3H) 2.93-3.09 (m, 6H) 3.40-3.69 (m, 8H) 4.11-4.40 (m, 3H) 4.43-4.81 (m, 3H) 4.99-5.17 (m, 2H) 6.92-7.44 (m, 14H) 7.46-7.64 (m, 4H) 7.75 (br. s., 3H) 8.01-8.22 (m, 2H) 8.01-8.06 (m, 1H)

[0392] Analogously the following compound has been prepared:

[0393] N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-chloromethyl)-1-methyl-6-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide

##STR00148##

[0394] ESI MS: m/z 1251 (MH+)

[0395] .sup.1H NMR (500 MHz, methanol-d.sub.4) ppm 0.94-1.01 (m, 6H) 1.51-1.95 (m, 4H) 2.12 (br. s., 5H) 2.48-2.62 (m, 3H) 2.89-3.05 (m, 4H) 3.40-3.54 (m, 4H) 3.46-3.65 (m, 4H) 3.58-3.74 (m, 2H) 3.75-4.05 (m, 1H) 4.08-4.42 (m, 3H) 4.50 (m, 1H) 4.55-4.81 (m, 4H) 4.96-5.20 (m, 2H) 6.92-7.81 (m, 17H) 8.04 (br. s., 1H)

Deprotection

[0396] L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(II)] (Compd 50)

##STR00149##

[0397] To a solution of N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methy)amino]ethyl}methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide (11 mg, 0.0078 mmol) in DMF (1 mL) was added piperidine (4 mL, 0.039 mmol). The mixture was stirred 1 hour at room temperature, solvent was evaporated and the resulting title compound was used without further purification.

[0398] Analogously the following compound has been prepared:

[0399] L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-chloromethyl)-1-methyl-6-({5-[2-pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(II)] (Compd 51)

##STR00150##

Step g

[0400] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8 dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd 52)

##STR00151##

[0401] To a solution of L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({(8S)-8-chloromethyl)-1-methyl-6-({5-[({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino)ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide (4.6 mg, 0.0039 mmol) in 1 mL of DCM (10% DMF), were added 1-{6-[(2,5-dioxopyrrolidin-1-yl)oxy]-6-oxohexyl}-1H-pyrrole-2,5-dione (3.6 mg, 0.0117 mmol) and triethylamine (2.2 mL, 0.0156 mmol). The mixture was stirred at room temperature for 3 hours, solvents were evaporated and the title compound (2.2 mg, 41% yield) was isolated by column chromatography purification (DCM/MeOH 8:2).

[0402] ESI MS: m/z 1380 (MH+)

[0403] .sup.1H NMR (500 MHz, methanol-d.sub.4) ppm 1.98 (br. s., 4H) 2.60-2.63 (m, 3H) 2.89-3.11 (m, 6H) 3.39-3.72 (m, 10H) 4.07-4.75 (m, 7H) 4.98-5.17 (m, 2H) 6.74 (br. s., 2H) 7.00 (m, 2H) 7.07-7.45 (m, 8H) 7.52 (br. s., 3H) 8.07-8.11 (m, 1H)

[0404] Analogously the following compounds have been prepared:

[0405] N-[19-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-17-oxo-4,7,10,13-tetraoxa-16-azanonadecan-1-oyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-6-({5-[({5-[2-pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)amino]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)] (Compd 53)

##STR00152##

[0406] ESI MS: m/z 1585 (MH+)

[0407] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[{2-[({[(8S)-8-(chloromethyl)-1-methyl-({5-[2-(pyrrolidin-1-yl)ethoxy]-1H-indol-2-yl}carbonyl)-7,8-dihydro-6H-thieno[3,2-e]indol-4-yl]oxy}carbonyl)(methyl)amino]ethyl}(methyl)carbamoyl]oxy}methyl)phenyl]-L-ornithinamide [(IV)](Compd 54)

##STR00153##

[0408] ESI MS: m/z 1222 (MH+)

[0409] .sup.1H NMR (400 MHz, dmf-d7) ppm 0.95 (t, J=7.8 Hz, 6H) 2.16 (m, 1H) 2.66 (br. s., 3H) 2.99-3.09 (m, 3H) 3.09-3.29 (m, 3H) 3.59 (br. s., 2H) 3.69 (br. s., 2H) 3.81 (br. s., 2H) 4.04 (d, J=10.7 Hz, 1H) 4.31-4.47 (m, 2H) 4.61 (br. s., 1H) 4.83 (br. s., 2H) 5.11 (d, J=15.7 Hz, 2H) 5.60 (s, 2H) 6.29 (br. s., 1H) 7.00 (m, 3H) 7.30 (m, 2H) 7.53 (d, J=8.5 Hz, 2H) 7.88 (d, J=8.2 Hz, 1H) 8.13 (d, J=7.8 Hz, 1H) 8.27 (s, 1H) 10.1 (m, 1H) 11.61 (br. s., 1H)

Preparation of the Intermediate

[0410] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-{4-[({[2,2-dimethyl-3-(methylamino)propyl] (methyl)carbamoyl}oxy)methyl]phenyl}-L-ornithinamide (XX)

##STR00154##

Step 1

N,N-(2,2-dimethylpropane-1,3-diyl)diformamide

[0411] ##STR00155##

[0412] In a round bottomed flask, commercially available 2,2-dimethylpropane-1,3-diamine (851 mg, 8.32 mmol) was reacted with ethylformiate (4.2 ml). The reaction mixture was stirred at 60 C. for 3 hours, until no starting material was detectable (TLC analysis, MeOH:CH.sub.2Cl.sub.2=2:8). The reaction mixture was then evaporated under vacuum, affording the crude product (1.4 g, oil).

[0413] ESI MS: m/z 159 (MH+)

[0414] .sup.1H NMR (600 MHz, DMSO-d.sub.6) ppm 0.79 (s, 6H) 2.92 (d, J=6.41 Hz, 4H) 7.94 (br. s., 2H) 8.05-8.09 (m, 2H)

Step 2

N,N,2,2-tetramethylpropane-1,3-diamine

[0415] ##STR00156##

[0416] In a dried round bottomed flask, containing N,N-(2,2-dimethylpropane-1,3-diyl)diformamide (1.4 g, 8.85 mmol) cooled at 0 C. under argon atmosphere, a solution (24 ml) of Lithium Aluminium Hydride in tetrahydrofuran (1M) was added. The reaction mixture was stirred at room temperature for 28 hours. After cooling at 0 C., a solution of water in tetrahydrofuran was added, and the resulting mixture was filtered on a celite pad. The filtrate was dried over anhydrous sodium sulfate, filtered and the product thus obtained as a tetrahydrofuran solution was used without further purification in the next step.

Step 3

2-(biphenyl-4-yl)propan-2-yl [2,2-dimethyl-3-(methylamino)propyl]methylcarbamate

[0417] ##STR00157##

[0418] Commercial methyl 4-[({[2-(biphenyl-4-yl)propan-2-yl]oxy}carbonyl)oxy]benzoate (5.54 mmol, 2.16 g) was added to the tetrahydrofuran solution of N,N,2,2-tetramethylpropane-1,3-diamine. The reaction mixture was stirred at room temperature for 24 hours. The solvent was then evaporated under vacuum and the residue was purified by flash chromatography (MeOH:CH.sub.2Cl.sub.2=1:9) affording the desired product (848 mg).

[0419] MS (ESI): 369 (MH+)

[0420] .sup.1H NMR (600 MHz, DMSO-d.sub.6) ppm 0.73-0.83 (m, 5H) 0.91 (br. s., 3H) 1.65-1.77 (m, 6H) 2.78 (br. s., 1H) 7.32-7.38 (m, 1H) 7.41 (d, J=8.43 Hz, 2H) 7.46 (t, J=7.69 Hz, 2H) 7.58-7.69 (m, 4H)

[0421] Analogously the following compound has been prepared:

[0422] 2-(biphenyl-4-yl)propan-2-yl methyl[2-(methylamino)ethyl]carbamate

##STR00158##

[0423] MS (ESI): 327 (MH+)

[0424] .sup.1H NMR (401 MHz, DMSO-d.sub.6) ppm 1.70-1.75 (m, 6H) 2.23-2.39 (m, 3H) 7.30-7.39 (m, 1H) 7.40-7.49 (m, 4H) 7.58-7.63 (m, 2H) 7.63-7.68 (m, 2H)

Step 4

N-[6-(2,5-dioxo-25-dihydro-1H-pyrrol-1-yl)hexanoyl]L-valyl-N-{4-[11-(biphenyl-4-yl)-4,6,6,8,11-pentamethyl-3,9-dioxo-2,10-dioxa-4,8-diazadodec-1-yl]phenyl}-N.SUP.5.-carbamoyl-L-ornithinamide

[0425] ##STR00159##

[0426] To a solution of N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-[4-({[(4-nitrophenoxy)carbonyl]oxy}methyl)phenyl]-L-omithinamide (0.492 mmol, 363 mg) in anhydrous dimethylsulfoxide (1.0 ml), a solution of 2-(biphenyl-4-yl)propan-2-yl [2,2-dimethyl-3-(methylamino)propyl]methylcarbamate (0.393 mmol, 145 mg) in anhydrous dimethylsulfoxide (0.9 ml) and triethylamine (1.5 mmol, 150 mg) were added. The reaction mixture was stirred at room temperature until no starting material was detectable, then treated with n-hexane (36 ml), and the crude residue was used without further purification in the next step.

[0427] MS (ESI): 967 (MH+).

[0428] Analogously the following compound has been prepared

[0429] N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-valyl-N-{4-[10-(biphenyl-4-yl)-4,7,10-trimethyl-3,8-dioxo-2,9-dioxa-4,7-diazaundec-1-yl]phenyl}N.sup.5-carbamoyl-L-ornithinamide

##STR00160##

[0430] MS (ESI): 954 (MH+)

[0431] .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 0.86 (dd, J=13.34, 6.79 Hz, 6H) 1.69 (br. s., 6H) 1.86-2.05 (m, 1H) 2.76 (m, J=9.30 Hz, 2H) 2.92 (d, J=14.95 Hz, 3H) 3.01 (m, J=6.10 Hz, 1H) 3.15-3.15 (m, 0H) 3.38-3.53 (m, 2H) 3.93 (t, J=7.63 Hz, 1H) 4.18-4.26 (m, 2H) 4.26-4.34 (m, 1H) 4.42 (br. s., 1H) 4.92-5.06 (m, 2H) 5.39 (s, 2H) 5.96 (t, J=6.02 Hz, 1H) 7.18-7.48 (m, 12H) 7.52-7.66 (m, 6H) 7.70-7.77 (m, 2H) 7.88 (d, J=7.47 Hz, 2H) 8.11 (br. s., 1H) 10.06 (br. s., 1H)

Step 5

The Title Intermediate

[0432] The crude product N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[11-(biphenyl-4-yl)-4,6,6,8,11-pentamethyl-3,9-dioxo-2, 10-dioxa-4,8-diazadodec-1-yl]phenyl}-N.sup.5-carbamoyl-L-ornithinamide was treated with dichloroacetic acid (7.8 mmol). The reaction mixture was stirred at room temperature, until no starting material was detectable, then purified by flash column chromatography (EtOH:CH.sub.2Cl.sub.2=1.5:8.5) on silica gel, affording the desired product (44.0 mg, white solid).

[0433] MS (ESI): 729 (MH+)

[0434] .sup.1H NMR (500 MHz, METHANOL-d.sub.4) ppm 0.97 (d, J=4.42 Hz, 3H) 0.98 (d, J=4.27 Hz, 3H) 1.04 (s, 6H) 1.23-1.35 (m, 4H) 1.50-1.68 (m, 7H) 1.76 (dtd, J=13.90, 9.26, 9.26, 5.11 Hz, 1H) 1.86-1.95 (m, 1H) 2.07 (dq, J=13.90, 6.86 Hz, 1H) 2.26-2.31 (m, 2H) 2.57 (s, 3H) 2.67 (s, 2H) 3.04 (s, 3H) 3.11 (dt, J=13.50, 6.67 Hz, 1H) 3.16-3.22 (m, 1H) 3.23 (br. s., 2H) 3.48 (t, J=7.09 Hz, 2H) 4.14 (d, J=7.47 Hz, 1H) 4.49 (dd, J=9.07, 5.11 Hz, 1H) 5.13 (s, 2H) 5.94 (s, 3H) 6.75-6.82 (m, 1H) 7.36 (d, J=8.54 Hz, 2H) 7.60 (d, J=8.69 Hz, 2H).

Preparation of Intermediate

[0435] N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N.sup.5-carbamoyl-N-{4-[({methyl[2-(methylamino)ethyl]carbamoyloxy}methyl]phenyl}-L-ornithinamide

##STR00161##

Step 6

N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[11-(biphenyl-4-yl)-4,6,6,8,11-pentamethyl-3,9-dioxo-2,10-dioxa-4,8-diazadodec-1-yl]phenyl}-N.SUP.5.-carbamoyl-L-omithinamide

[0436] ##STR00162##

[0437] To a solution of the product N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-val-N-{4-[10-(biphenyl-4-yl)-4,7,10-trimethyl-3,8-dioxo-2,9-dioxa-4,7-diazaundec-1yl]phenyl}-N.sup.5-carbamoyl-L-ornithinamide [prepared as reported above under step 4] (0.063 mmol, 60 mg) in anhydrous dimethylformamide (0.8 ml), piperidine (0.32 mmol, 27.2 mg) was added. The reaction mixture was stirred at room temperature until no starting material was detectable. The solvent was evaporated under vacuum and the crude thus obtained was used without further purification in the next step.

[0438] MS (ESI): 732 (MH+)

Step 7

N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[10-(biphenyl-4-yl)-4,7,10-trimethyl-3,8-dioxo-2,9-dioxa-4,7-diazaundec-1-yl]phenyl}-N.SUP.5.-carbamoyl-L-omithinamide

[0439] ##STR00163##

[0440] To a solution of the crude product N-[6-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[11-(biphenyl-4-yl)-4,6,6,8,11-pentamethyl-3,9-dioxo-2,10-dioxa-4,8-diazadodec-1-yl]phenyl}-N.sup.5-carbamoyl-L-omithinamide (0.0629 mmol) in anhydrous dimethylformamide (1.5 ml), 1-{6-[(2,5-dioxopyrrolidin-1-yl)oxy]-6-oxohexyl}-1H-pyrrole-2,5-dione (0.189 mmol, 58.3 mg) and triethylamine (0.252 mmol, 25.5 mg) were added. The reaction mixture was stirred at room temperature until no starting material was detectable. The solvent was evaporated under vacuum and the crude was purified by flash column chromatography (eluant:EtOH:CH.sub.2Cl.sub.2=1:9) on silica gel (230-400 mesh), affording the desidered product (34 mg, white wax)

[0441] MS (ESI): 925 (MH+)

[0442] .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 0.83 (dd, J=15.79, 6.79 Hz, 6H) 1.31-1.40 (m, 1H) 1.60 (br. s., 1H) 1.69 (s, 6H) 1.90-2.02 (m, 1H) 2.05-2.23 (m, 2H) 2.89-2.96 (m, 3H) 3.01 (br. s., 1H) 3.34-3.38 (m, 2H) 3.46 (m, J=8.24 Hz, 2H) 4.19 (t, J=7.40 Hz, 1H) 4.38 (br. s., 1H) 4.95-5.05 (m, 2H) 5.40 (s, 2H) 5.97 (d, J=12.20 Hz, 1H) 6.98-7.02 (m, 2H) 7.30 (d, J=8.85 Hz, 2H) 7.37 (dd, J=15.40, 7.93 Hz, 3H) 7.45 (m, J=7.17 Hz, 2H) 7.52-7.67 (m, 6H) 7.80 (d, J=8.24 Hz, 1H) 8.08 (d, J=6.71 Hz, 1H) 9.76-10.18 (m, 1H)

Step 8

The Title Intermediate

[0443] Reacting the intermediate prepared under step 6 under reaction conditions reported above in step 5, the title compound as a white powder was obtained

##STR00164##

[0444] MS (ESI): 687 (MH+)