Patent EA-002066-B1 (EA)
Title: Conjugates useful in the treatment of prostate cancer
Inventor: GARSKY VICTOR M
IPC: A61K47/48
Filed: 1997-10-27
Abstract: 1. A conjugate which is useful for the treatment of prostate cancer which comprises a cytotoxic agent attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is selectively proteolytically cleaved by free prostate specific antigen and wherein the means of attachment is a covalent bond or through a chemical linker, said sequence of amino acids which comprises at least one cyclic amino acid having a hydrophilic substituent; or the pharmaceutically acceptable salt thereof. 2. The conjugate according to Claim 1 wherein the cytotoxic agent is a member of a class of cytotoxic agents selected from the following classes: a) anthracycline family of drugs, b) the vinca alkaloid drugs, c) the mitomycins, d) the bleomycins, e) the cytotoxic nucleosides, f) the pteridine family of drugs, g) diynenes, h) estramustine, i) cyclophosphamide, j) the taxanes and k) the podophyllotoxins, or the pharmaceutically acceptable salt thereof. 3. The conjugate according to Claim wherein the cytotoxic agent is selected from the following cytotoxic agents: a) doxorubicin, b) carminomycin, <DP=2 c) daunorubicin, d) aminopterin, e) methotrexate, f) methopterin, g) dichloro-methotrexate, h) mitomycin C, i) porfiromycin, j) 5-fluorouracil, k) 6-mercaptopurine, l) cytosine arabinoside, m) podophyllotoxin, n) etoposide, o) etoposide phosphate, p) melphalan, q) vinblastine, r) vincristine, s) leurosidine, t) vindesine, u) estramustine, v) cisplatin, w) cyclophosphamide, x) taxol, and y) leurosine, or the pharmaceutically acceptable salt thereof. 4. The conjugate according to Claim 2 wherein the cytotoxic agent is selected from doxorubicin and vinblastine or a cytotoxic derivative thereof. 5. The conjugate according to Claim 2 wherein the cytotoxic agent is doxorubicin or a cytotoxic derivative thereof. 6. The conjugate according to Claim 1 wherein the oligopeptide comprises an oligomer selected from: a) HaaXaaSerTyrGlnISerSer (SEQ.ID.NO.: 1); b) HaaTyrGlnISerSer (SEQ.ID.NO.:2); c) HaaXaaLysTyrGlnISerSer (SEQ.ID.NO.:3); d) HaaXaaLysTyrGlnISerSer (SEQ.ID.NO.:4); e) HaaXaahArgTyrGlnISerSer (SEQ.ID.NO.:5); f) HaaXaahArgChaGlnISerSer (SEQ.ID.NO.: 6); g) HaaXaaSerTyrGlnISerXaa (SEQ.ID.NO.: 7); h) HaaTyrGInISerXaa (SEQ.ID.NO.: 8); i) HaaXaaSerChgGlnISerXaa (SEQ.ID.NO.: 9); j) HaaChgGInISerXaa (SEQ.ID.NO.: 10);  wherein Haa is a cyclic amino acid substituted with a hydrophilic moiety, Xaa is any amino acid, hArg is homoarginine, Cha is cyclohexylalanine and Chg is cyclohexylglycine. 7. The conjugate according to Claim 1 wherein the oligopeptide comprises an oligomer selected from: a) HaaXaaSerTyrGlnISerSer (SEQ.ID.NO.:11), b) HaaXaaSerTyrGlnISerAla (SEQ.ID.NO.: 12), <DP=4 c) AlaHaaXaaSerTyrTyrISer (SEQ.ID.NO.: 13), d) AlaAsnHaaXaaSerTyrGlnISer (SEQ.ID.NO.: 14), e) HaaXaaSerTyrGlnISerSerThr (SEQ.ID.NO.: 15), HaaTyrGlnISerSerThr (SEQ.ID.NO.:16), g) HaaXaaSerTyrGlnISerSerSer (SEQ.ID.NO.: 17), h) HaaTyrGlnISerSerSer (SEQ.ID.NO:: 18), i) HaaXaaLysTyrGlnISerSerSer (SEQ.ID.NO.: 19), j) HaaXaahArgTyrGlnISerSerSer (SEQ.ID.NO.:20), k) HaaXaaSerTyrGlnISerSerLeu (SEQ.ID.NO.:21); l) HaaTyrGlnISerSerLeu (SEQ.ID.NO.:22); m) HaaXaaSerTyrGlnISerLeu (SEQ.ID.NO.: 23); n) HaaTyrGlnlSerLeu (SEQ.ID.NO.: 24); p) HaaXaaSerTyrGlnISerNle (SEQ.ID.NO.: 25); q) HaaTyrGlnISerNle (SEQ.ID.NO.: 26); r) HaaXaaSerTyrGlnISerTIC (SEQ.ID.NO.: 27); s) HaaTyrGlnISerTIC (SEQ.ID.NO.: 28); t) HaaXaaSerChgGlnISerLeu (SEQ.ID.NO.: 29); <DP=5 u) HaaChgGlnISerLeu (SEQ.ID.NO.: 30); v) HaaXaaSerChgGlnISerNle (SEQ.ID.NO.: 31); w) HaaChgGlnISerNle (SEQ.ID.NO.: 32); x) HaaXaaSerChgGlnlSerTIC (SEQ.ID.NO.: 33); y) HaaChgGlnISerTIC (SEQ.ID.NO.: 34); z) hArgChgGlnISerLeu (SEQ.ID.NO.: 35); and aa) hArgTyrGlnISerLeu (SEQ.ID.NO.: 36). 8. The conjugate according to Claim 1 wherein the oligopeptide comprises an oligomer selected from: a) 4-HypXaaSerTyrGlnISerSer (SEQ.ID.NO.:37), b) 4-HypXaaSerTyrGlnISerAla (SEQ.ID.NO.: 38), c) Ala4-HypXaaSerTyrTyrISer (SEQ.ID.NO.:39), d) AlaAsn4-HypXaaSerTyrGlnISer (SEQ.ID.NO.:40), e) 4-HypXaaSerTyrGlnISerSerThr (SEQ.ID.NO.:41), f) 4-HypTyrGlnISerSerThr (SEQ.ID.NO.:42), g) 4-HypXaaSerTyrGlnISerSerSer (SEQ.ID.NO.:43), h) 4-HypTyrGlnISerSerSer (SEQ.ID.NO.:44), i) 4-HypXaaLysTyrGlnISerSerSer (SEQ.ID.NO.:45), <DP=6 j) 4-HypXaahArgTyrGlnISerSerSer (SEQ.ID.NO.:46), k) 4-HypXaaSerTyrGlnlSerSerLeu (SEQ.ID.NO.:47); l) 4-HypTyrGlnISerSerLeu (SEQ.ID.NO.:48); m) 4-HypXaaSerTyrGlnISerLeu (SEQ.ID.NO.:49); n) 4-HypTyrGlnISerLeu (SEQ.ID.NO.: 50); p) 4-HypXaaSerTyrGlnISerNle (SEQ.ID.NO.:51); q) 4-HypTyrGlnISerNle (SEQ.ID.NO.: 52); r) 4-HypXaaSerTyrGlnISerTIC (SEQ.ID.NO.: 53); s) 4-HypTyrGlnISerTIC (SEQ.ID.NO.: 54); t) 4-HypXaaSerChgGlnISerLeu (SEQ.ID.NO.: 55); u) 4-HypChgGlnISerLeu (SEQ.ID.NO.: 56); v) 4-HypXaaSerChgGlnISerNle (SEQ.ID.NO.: 57); w) 4-HypChgGlnISerNle (SEQ.ID.NO.: 58); x) 4-HypXaaSerChgGlnISerTIC (SEQ.ID.NO.: 59); y) 4-HypChgGlnISerTIC (SEQ.ID.NO.: 60);] wherein 4-Hyp is 4-hydroxyproline, Xaa is any amino acid, hArg is homoarginine, Cha is cyclohexylalanine and Chg is cyclohexylglycine. 9. The conjugate according to Claim 1 wherein the cyclic amino acid having a hydrophilic substituent is selected from:   wherein: R<5> is selected from HO- and C1-C6 alkoxy; R<6> is selected from hydrogen, halogen, C1-C6 alkyl, HO- and C1-C6 alkoxy; and t is 3 or 4. 10. A conjugate which is useful for the treatment of prostate cancer of the formula I:   wherein: oligopeptide is an oligopeptide which is selectively recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen, wherein the oligopeptide comprises a cyclic amino acid of the formula:   and wherein the C-terminus carbonyl is covalently bound to the amine of doxorubicin; R is selected from a) hydrogen, b) -(C=O)R<1a> c)    d)    e)    R<1> and R<2> are independently selected from: hydrogen, OH, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 aralkyl and aryl; R<1a> is C1-C6-alkyl, hydroxylated aryl, polyhydroxylated aryl or aryl, R<5> is selected from HO- and C1-C6 alkoxy; R<6> is selected from hydrogen, halogen, C1-C6 alkyl, HO- and C1-C6 alkoxy; and n is 1, 2, 3 or 4; p is zero or an integer between 1 and 100; q is 0 or 1, provided that if p is zero, q is 1; r is an integer between 1 and 10; and t is 3 or 4; or a pharmaceutically acceptable salt thereof. 11. The conjugate according to Claim 10 wherein the cyclic amino acid is    R is selected from a) hydrogen, b) -(C=O)R<1a>, c)    d).   e)    f)    g).   R<1> and R<2> are independently selected from: hydrogen, C1-C6 alkyl and aryl; R<1a> is C1-C6-alkyl or aryl, n is 1, 2, 3 or 4; n' is 0, 1, 2 or 3; p is zero or an integer between 1 and 14; q is 0 or 1, provided that if p is zero, q is 1; r is an integer between 1 and 10; t is 3; or a optical isomer or pharmaceutically acceptable salt thereof. 12. The conjugate according to Claim 10 wherein: oligopeptide is an oligomer that comprises an amino acid sequence selected from: a) 4-HypXaaSerTyrGlnISerSer (SEQ.ID.NO.:37), b) 4-HypXaaSerTyrGlnISerAla (SEQ.ID.NO.:38), c) Ala-4-HypXaaSerTyrTyrISer (SEQ.ID.NO.:39), d) AlaAsn4-HypXaaSerTyrGlnISer (SEQ.ID.NO.:40), e) 4-HypXaaSerTyrGlnISerSerThr (SEQ.ID.NO.:41), f) 4-HypTyrGlnISerSerThr (SEQ.ID.NO.:42), g) 4-HypXaaSerTyrGlnISerSerSer (SEQ.ID.NO.:43), h) 4-HypTyrGlnISerSerSer (SEQ.ID.NO.:44), i) 4-HypXaaLysTyrGlnISerSerSer (SEQ.ID.NO.:45), <DP=12 j) 4-HypXaahArgTyrGlnISerSerSer (SEQ.ID.NO.:46), k) 4-HypXaaSerTyrGlnISerSerLeu (SEQ.ID.NO.:47); l) 4-HypTyrGlnISerSerLeu (SEQ.ID.NO.:48); m) 4-HypXaaSerTyrGlnISerLeu (SEQ.ID.NO.:49); n) 4-HypTyrGlnISerLeu (SEQ.ID.NO.:50); p) 4-HypXaaSerTyrGlnISerNle (SEQ.ID.NO.:51); q) 4-HypTyrGlnISerNle (SEQ.ID.NO.:52); r) 4-HypXaaSerTyrGlnISerTIC (SEQ.ID.NO.:53); s) 4-HypTyrGlnISerTIC (SEQ.ID.NO.:54); t) 4-HypXaaSerChgGlnISerLeu (SEQ.ID.NO.: 55); u) 4-HypChgGlnISerLeu (SEQ.ID.NO.:56); v) 4-HypXaaSerChgGlnISerNle (SEQ.ID.NO.:57); w) 4-HypChgGlnISerNle (SEQ.ID.NO.:58); x) 4-HypXaaSerChgGlnISerTIC (SEQ.ID.NO.:59); y) 4-HypChgGlnISerTIC (SEQ.ID.NO.:60); wherein 4-Hyp is 4-hydroxyproline, Xaa is any amino acid, hArg is homoarginine, Cha is cyclohexylalanine and Chg is cyclohexylglycine; or an optical isomer or pharmaceutically acceptable salt thereof. 13. The conjugate according to Claim 12 wherein: Xaa is alanine or isoleucine; or an optical isomer or pharmaceutically acceptable salt thereof. 14. The conjugate according to Claim 10 which is selected from:   wherein X is:     Succinyl-(4-Hyp)ASChgQ-SV-DOX (3') 75 Glutaryl-(4-Hyp)ASChgQ-SV-DOX (3') 75 Glutaryl-(4-Hyp)ASChgQ-SI-DOX (3') 77 Succinyl-(4-Hyp)SSChgQ-SI-DOX (3') 78 <DP=15 Succinyl-(4-Hyp)ASChgQ-SI-DOX (3') 79 Succinyl-(4-Hyp)ASChgQ-SAbu-DOX (3') 80 Glutaryl-(4-Hyp)SSChgQ-SI-DOX (3') 81 Glutaryl-(4-Hyp)SSChgQ-SL-DOX (3') 82 PEG(2)-(4-Hyp)SSChgQ-SL-DOX (3') 83 Succinyl-(4-Hyp)ASChgQ-SThi-DOX (3') 84 PEG(4)-(4-Hyp)-SSChgQ-SL-DOX (3') 85 PEG 2)-(4-Hyp)ASChgQ-SThi-DOX (3') 86 Succinyl-3,4-(diOH)PASChgQ-SL-DOX (3') 87 Malonyl-(4-Hyp)ASChgQ-SL-DOX (3') 88 or an optical isomer or pharmaceutically acceptable salt thereof. 15. The conjugate according to Claim 10 which is: SEQ. ID. NO. Succinyl-(4-trans-L-Hyp)ASChgQ-SV-DOX (3') 75 Glutaryl-(4-trans-L-Hyp)ASChgQ-SV-DOX (3') 76 Glutaryl-(4-trans-L-Hyp)ASChgQ-SI-DOX (3') 77 Succinyl-(4-trans-L-Hyp)SSChgQ-SI-DOX (3') 78 Succinyl-(4-trans-L-Hyp)ASChgQ-SI-DOX (3') 79 Succinyl-(4-trans-L-H)ASChgQ-SAbu-DOX (3') 80 Glutaryl-(4-trans-L-Hyp)SSChgQ-SI-DOX (3') 81 Glutaryl-(4-trans-L-Hyp)SSChgQ-SL-DOX (3') 82 PEG(2)-(4-trans-L-Hyp)SSChgQ-SL-DOX (3') 83 Succinyl-(4-trans-L-Hyp)ASChgQ-SThi-DOX (3') 84 PEG(4)-(4-)trans-L-Hyp)-SSChgQ-SL-DOX (3') 85 PEG(2)-(4-)trans-L-Hyp)ASChgQ-SThi-DOX (3') 86 Succinyl-3,4-(diOH)PASChgQ-SL-DOX (3') 87 Malonyl-(4-trans-L-Hyp)ASChgQ-SL-DOX (3') 88 or an optical isomer or pharmaceutically acceptable salt thereof. 16. The conjugate according to Claim 10 which is: [N-Ac-(4-trans-L-Hyp)]-Ala-Ser-Chg-Gln-Ser-Leu-Dox    or an optical isomer or