Conjugates useful in the treatment of prostate cancer

• US 5,948,750 A
• Filed: 10/14/1997
• Issued: 09/07/1999
• Est. Priority Date: 10/30/1996
• Status: Expired due to Fees

• Current Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Original Assignee: Merck & Co., Inc.
• Inventors: Defeo-Jones, Deborah, Garsky, Victor M., Feng, Dong-Mei
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US08/950,805
• Time in Patent Office: 693 Days
• Field of Search: 514/2, 514/8, 514/16, 514/17, 514/18, 514/34, 514/283, 530/300, 530/322, 530/328, 530/329, 530/330, 530/345, 536/6.4, 536/17.2, 540/478
• US Class Current: 514/19.5
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: C07K 14/47 from mammals

Abstract

Chemical conjugates which comprise oligopeptides, having amino acid sequences that are selectively proteolytically cleaved by free prostate specific antigen (PSA) and known cytotoxic agents are disclosed. Such conjugates are useful in the treatment of prostatic cancer and benign prostatic hypertrophy (BPH).

Claims

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 1 wherein the cytotoxic agent is selected from the following cytotoxic agents:

a) doxorubicin,

b) carminomycin,

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, vinblastine and desacetylvinblastine.

5. The conjugate according to claim 2 wherein the cytotoxic agent is doxorubicin.

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) HaaTyrGlnISerXaa (SEQ.ID.NO.;

     8);

i) HaaXaaSerChgGlnISerXaa (SEQ.ID.NO.;

     9);

j) HaaChgGlnISerXaa (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.;

     1),

b) HaaXaaSerTyrGlnISerAla (SEQ.ID.NO.;

     12),

c) AlaHaaXaaSerTyrTyrISer (SEQ.ID.NO.;

     13),

d) AlaAsnHaaXaaSerTyrGlnISer (SEQ.ID.NO.;

     14),

e) HaaXaaSerTyrGlnISerSerThr (SEQ.ID.NO.;

     15),

f) 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) HaaTyrGlnISerLeu (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);

u) HaaChgGlnISerLeu (SEQ.ID.NO.;

     30);

v) HaaXaaSerChgGlnISerNle (SEQ.ID.NO.;

     31);

w) HaaChgGlnISerNle (SEQ.ID.NO.;

     32);

x) HaaXaaSerChgGlnISerTIC (SEQ.ID.NO.;

     33);

y) HaaChgGlnISerTIC (SEQ.ID.NO.;

     34);

z) HaahArgChgGlnISerLeu (SEQ.ID.NO.;

     35); and

aa) HaahArgTyrGlnISerLeu (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),

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.

9. The conjugate according to claim 1 wherein the cyclic amino acid having a hydrophilic substituent is selected from:

##STR51## wherein;

R⁵ is selected from HO-- and C1-C6 alkoxy;

R6 is selected from hydrogen, halogen, C1-C6 alkyl, HO-- and C1-C6 alkoxy; and

t is 3 or 4.

19. The conjugate according to claim 1 of the formula II:

##STR68## wherein;

oligopeptide is an oligopeptide which is specifically 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, and the oligopeptide comprises a cyclic amino acid of the formula;

##STR69## XL is --NH--(CH2)u--NH--R is selected from ##STR70## R1 and R2 are independently selected from;

hydrogen, OH, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 aralkyl and aryl;

R1a is C1-C6-alkyl, hydroxylated aryl, polyhydroxylated aryl or aryl,

R⁵ is selected from HO-- and C₁ -C₆ alkoxy;

R⁶ is selected from hydrogen, halogen, C₁ -C₆ alkyl, HO- and C₁ -C₆ alkoxy;

R19 is hydrogen, (C1-C3 alkyl)-CO, or chlorosubstituted (C1-C3 alkyl)-CO;

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 1, 2 or 3;

t is 3 or 4;

u is 1, 2, 3, 4 or 5,

or a pharmaceutically acceptable salt thereof.

20. The conjugate according to claim 19 which is selected from:

##STR71## or a pharmaceutically acceptable salt or optical isomer thereof.

21. The conjugate according to claim 1 of the formula ##STR72## wherein:

oligopeptide is an oligopeptide which is specifically 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, and the oligopeptide comprises a cyclic amino acid of the formula;

##STR73## R⁵ is selected from HO-- and C₁ -C₆ alkoxy;

R⁶ is selected from hydrogen, halogen, C₁ -C₆ alkyl, HO-- and C₁ -C₆ alkoxy;

Rg and R^h are independently selected from;

hydrogen, C₁ -C₆ -alkyl, -C₁ -C₆ -alkyl-OH, -C₁ -C₆ -alkyl-di-OH, -C₁ -C₆ -alkyl-tri-OH and ##STR74## provided that at least one R^g and R^h are not hydrogen or C₁ -C₆ -alkyl, or

Rg and R^h are combined to form a --CH₂ CH₂ OCH₂ CH₂ -- diradical;

R¹⁹ is hydrogen, (C₁ -C₃ alkyl)-CO, or chloro-substituted (C₁ -C₃ alkyl)-CO;

p is zero or an integer between 1 and 100;

q is 0 or 1, provided that if p is zero, q is 1;

t is 3 or 4

or a pharmaceutically acceptable salt thereof.

22. The conjugate according to claim 21 which is:

##STR75## or a pharmaceutically acceptable salt or optical isomer thereof.

23. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate of claim 1.

27. A method for treating prostate cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 23.

31. A method for treating benign prostatic hyperplasia which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 23.

35. A pharmaceutical composition made by combining the conjugate of claim 1 and a pharmaceutically acceptable carrier.

36. A process for making a pharmaceutical composition comprising combining a conjugate of claim 1 and a pharmaceutically acceptable carrier.

37. The conjugate according to claim 2 wherein the cytotoxic agent is selected from a group consisting of an anthracycline antibiotic agent and a vinca drug.

38. The conjugate according to claim 2 wherein the cytotoxic agent is an anthracycline antibiotic agent.

10. A conjugate which is useful for the treatment of prostate cancer of the formula I:

##STR52## 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;

##STR53## and wherein the C-terminus carbonyl is covalently bound to the amine of doxorubicin;

R is selected from ##STR54## R¹ and R² 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,

R⁵ is selected from HO-- and C1-C6 alkoxy;

R⁶ 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 ##STR55## R is selected from ##STR56## R¹ and R² 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),

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:

##STR57## wherein X is;

space="preserve" listing-type="tabular">__________________________________________________________________________ ##STR58## 1 (SEQ. ID. NO.;

     68), 2 #STR59## (SEQ. ID. NO.;

     69), 3 #STR60## (SEQ. ID. NO.;

     70), 4 #STR61## (SEQ. ID. NO.;

     71), 5 #STR62## (SEQ. ID. NO.;

     72), 6 #STR63## (SEQ. ID. NO.;

     73), 7 #STR64## (SEQ. ID. NO.;

     74), __________________________________________________________________________ SEQ. ID. NO. __________________________________________________________________________ Succinyl-(4-Hyp)ASChgQ-SV-DOX(3'"'"') 75 Glutaryl-(4-Hyp)ASChgQ-SV-DOX(3'"'"') 76 Glutaryl-(4-Hyp)ASChgQ-SI-DOX(3'"'"') 77 Succinyl-(4-Hyp)SSChgQ-SI-DOX(3'"'"') 78 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:

space="preserve" listing-type="tabular">______________________________________ 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-Hyp)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 ##STR65## or an optical isomer or pharmaceutically acceptable salt thereof.

17. The conjugate according to claim 10 which is:

##STR66## or an optical isomer or pharmaceutically acceptable salt thereof.

18. The conjugate according to claim 10 which is:

##STR67## or an optical isomer or pharmaceutically acceptable salt thereof.

24. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate of claim 10.

25. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate of claim 14.

26. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate of claim 17.

28. A method for treating prostate cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 24.

29. A method for treating prostate cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 25.

30. A method for treating prostate cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 26.

32. A method for treating benign prostatic hyperplasia which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 24.

33. A method for treating benign prostatic hyperplasia which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 25.

34. A method for treating benign prostatic hyperplasia which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of claim 26.