Cell growth inhibitory substance

• US 4,816,444 A
• Filed: 07/10/1987
• Issued: 03/28/1989
• Est. Priority Date: 07/10/1987
• Status: Expired due to Term

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Pettit, George R., Kamano, Yoshiaki, Herald, Cherry L.
• Primary Examiner(s): Gerstl, Robert
• Application Number:
  US07/071,924
• Time in Patent Office: 627 Days
• Field of Search: 548/204, 514/17, 514/19, 530/330
• US Class Current: 514/19.3
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 5/0205 containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof

Abstract

A potent cell growth inhibitory substance obtained from the Indian Ocean sea hare Dolabella, herein denominated "dolastatin 10"; and pharmaceutical preparations containing same; and methods useful for the treatment of a host afflicted with neoplastic disease therewith.

Claims

1. A cell growth inhibitory substance denominated dolastatin 10 and having the structural formula:

##STR5##

3. A cell growth inhibitory substance according to claim 1 having the following N.M.R. characteristics:

space="preserve" listing-type="tabular">______________________________________ Chemical Shift, ppm Structure .sup.1 H(mult;

J,Hz;

Assignment 13C(mult) Integration) NOE ______________________________________ 2 170.51(s) 7.717(d;

3.3;

1H) 4 142.77(d) 7.254(d;

3.3;

1H) 5 118.76(d) 5.516(ddd;

5.7,7.2, 9.3;

1H) 6 53.02(d) 3.399(dd;

5.7,14;

1H) 6a 41.48(t) 3.256(dd;

5.7;

14;

1H) 6b1 137.74(s) 7.243(d;

7.9;

2H) 6b2,6b6 128.74(d)×

2 7.214(dd;

7.9,9.2;

2H) 6b3,6b5 129.80(d)×

2 7.194(t;

9.2;

1H) 6b4 127.02(d) 7.256(t;

7.6;

1H) 7 9,9a,10, 10ab,11 8 175.67(s) 2.282(quintet;

7.2;

1H) 7 9 44.79(d) 1.085(d;

7.1;

3H) 9a 14.49(q) 3.845(dd;

2.0,8.2;

1H) 7 10 82.05(d) 3.309(s;

3H) 6,7 10ab 60.89(q) 3.985(m;

1H) 11 59.86(d) 1.804(ddd;

5.5,7.0;

19;

1H) 12 25.00(t) 1.608(ddd;

7;

9.2,19;

1H) 1.446(ddd;

4.7,7,19;

1H) 13 25.45(t) 1.715(ddd;

4.7,7.8,12.7;

1H) 3.401(dd;

7.8,10;

1H) 17 14 48.03(t) 3.390(m;

1H).sup.b 16 174.01(s) 17 38.11(t) 2.394(ABq;

9.0;

2H) 14 18 78.86(d) 4.122(broad t;

8.7;

1H) 18ab 58.16(q) 3.313(s;

3H) 22 19 54.11(d) 3.26-3.39(1H).sup.c 19a 33.62(d) 1.680(1H).sup.b 19b 26.25(t) 1.370(broad m;

1H) 1.000(broad m;

1H).sup.b 19c 10.92(q) 0.823(t;

7.4;

3H) 19d 19.82(q) 1.003(d;

6.8;

3H) 20a 30.09(q) 3.012(s;

3H) 22 21 171.39(s) 22 54.20(d) 4.761(dd;

6.5, 8.8;

1H) 18,18ab, 20,19 22a 31.42(d) 1.983(sextet;

6.7;

1H) 22b 18.18(q) 0.941(d;

6.8;

3H) 22c 16.09(q) 0.977(d;

6.8;

3H) 23 6.861(d;

8.9;

1H) 25bc 24 172.44(s) 25 76.77(d) 2.454(d;

6.9;

1H) 23 25bc 49.92(q)×

2 2.262(s;

6H) 23 26 28.08(d) 2.073(sextet;

6.7;

1H) 27 20.24(q) 0.964(d;

6.8;

3H) 28 18.18(q) 0.902(d;

6.8;

3H) ______________________________________ .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm). .sup.b Overlapping signal. .sup.c Signal assigned from NOE data.

2. A pharmaceutical preparation comprising a pharmaceutically acceptable carrier and an effective amount of a natural or synthetic cell growth inhibitory substance or a non-toxic pharmaceutical active derivative thereof, said substance having the structural formula:

##STR6##

4. A pharmaceutical preparation according to claim 2 in which said substance has the following N.M.R. characteristics:

space="preserve" listing-type="tabular">______________________________________ Chemical Shift, ppm Structure .sup.1 H(mult;

J,Hz;

Assignment 13C(mult) Integration) NOE ______________________________________ 2 170.51(s) 7.717(d;

3.3;

1H) 4 142.77(d) 7.254(d;

3.3;

1H) 5 118.76(d) 5.516(ddd;

5.7,7.2, 9.3;

1H) 6 53.02(d) 3.399(dd;

5.7,14;

1H) 6a 41.48(t) 3.256(dd;

5.7;

14;

1H) 6b1 137.74(s) 7.243(d;

7.9;

2H) 6b2,6b6 128.74(d)×

2 7.214(dd;

7.9,9.2;

2H) 6b3,6b5 129.80(d)×

2 7.194(t;

9.2;

1H) 6b4 127.02(d) 7.256(t;

7.6;

1H) 7 9,9a,10, 10ab,11 8 175.67(s) 2.282(quintet;

7.2;

1H) 7 9 44.79(d) 1.085(d;

7.1;

3H) 9a 14.49(q) 3.845(dd;

2.0,8.2;

1H) 7 10 82.05(d) 3.309(s;

3H) 6,7 10ab 60.89(q) 3.985(m;

1H) 11 59.86(d) 1.804(ddd;

5.5,7.0;

19;

1H) 12 25.00(t) 1.608(ddd;

7;

9.2,19;

1H) 1.446(ddd;

4.7,7,19;

1H) 13 25.45(t) 1.715(ddd;

4.7,7.8,12.7;

1H) 3.401(dd;

7.8,10;

1H) 17 14 48.03(t) 3.390(m;

1H).sup.b 16 174.01(s) 17 38.11(t) 2.394(ABq;

9.0;

2H) 14 18 78.86(d) 4.122(broad t;

8.7;

1H) 18ab 58.16(q) 3.313(s;

3H) 22 19 54.11(d) 3.26-3.39(1H).sup.c 19a 33.62(d) 1.680(1H).sup.b 19b 26.25(t) 1.370(broad m;

1H) 1.000(broad m;

1H).sup.b 19c 10.92(q) 0.823(t;

7.4;

3H) 19d 19.82(q) 1.003(d;

6.8;

3H) 20a 30.09(q) 3.012(s;

3H) 22 21 171.39(s) 22 54.20(d) 4.761(dd;

6.5, 8.8;

1H) 18,18ab, 20,19 22a 31.42(d) 1.983(sextet;

6.7;

1H) 22b 18.18(q) 0.941(d;

6.8;

3H) 22c 16.09(q) 0.977(d;

6.8;

3H) 23 6.861(d;

8.9;

1H) 25bc 24 172.44(s) 25 76.77(d) 2.454(d;

6.9;

1H) 23 25bc 49.92(q)×

2 2.262(s;

6H) 23 26 28.08(d) 2.073(sextet;

6.7;

1H) 27 20.24(q) 0.964(d;

6.8;

3H) 28 18.18(q) 0.902(d;

6.8;

3H) ______________________________________ .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm). .sup.b Overlapping signal. .sup.c Signal assigned from NOE data.

5. A method of treating a host afflicted with neoplastic disease comprising administering to said host an effective amount of a natural or synthetic cell growth inhibitory substance or a pharmaceutically active non-toxic derivative thereof, said substance having the structural formula:

##STR7##

6. A method according to claim 5 in which said substance is administered intravenously, at a dosage level of from 0.1 up to about 20 mg per kilogram of host body weight.

7. A method according to claim 5 in which said substance is administered subcutaneously, at a dosage level of from 1 up to about 50 mg per kilogram of host body weight.

8. A method according to claim 5 in which said substance is administered orally, at a dosage level of from 5 up to about 100 mg per kilogram of host body weight.

9. A method according to claim 5 in which said neoplastic disease is lymphocytic leukemia P388.

10. A method according to claim 9 in which said small but effective amount comprises from about 1 up to about 4 μ

g per kilogram of host body weight.

11. A method according to claim 5 in which said substance has the following N.M.R. characteristics:

space="preserve" listing-type="tabular">______________________________________ Chemical Shift, ppm Structure .sup.1 H(mult;

J,Hz;

Assignment 13C(mult) Integration) NOE ______________________________________ 2 170.51(s) 7.717(d;

3.3;

1H) 4 142.77(d) 7.254(d;

3.3;

1H) 5 118.76(d) 5.516(ddd;

5.7,7.2, 9.3;

1H) 6 53.02(d) 3.399(dd;

5.7,14;

1H) 6a 41.48(t) 3.256(dd;

5.7;

14;

1H) 6b1 137.74(s) 7.243(d;

7.9;

2H) 6b2,6b6 128.74(d)×

2 7.214(dd;

7.9,9.2;

2H) 6b3,6b5 129.80(d)×

2 7.194(t;

9.2;

1H) 6b4 127.02(d) 7.256(t;

7.6;

1H) 7 9,9a,10, 10ab,11 8 175.67(s) 2.282(quintet;

7.2;

1H) 7 9 44.79(d) 1.085(d;

7.1;

3H) 9a 14.49(q) 3.845(dd;

2.0,8.2;

1H) 7 10 82.05(d) 3.309(s;

3H) 6,7 10ab 60.89(q) 3.985(m;

1H) 11 59.86(d) 1.804(ddd;

5.5,7.0;

19;

1H) 12 25.00(t) 1.608(ddd;

7;

9.2,19;

1H) 1.446(ddd;

4.7,7,19;

1H) 13 25.45(t) 1.715(ddd;

4.7,7.8,12.7;

1H) 3.401(dd;

7.8,10;

1H) 17 14 48.03(t) 3.390(m;

1H).sup.b 16 174.01(s) 17 38.11(t) 2.394(ABq;

9.0;

2H) 14 18 78.86(d) 4.122(broad t;

8.7;

1H) 18ab 58.16(q) 3.313(s;

3H) 22 19 54.11(d) 3.26-3.39(1H).sup.c 19a 33.62(d) 1.680(1H).sup.b 19b 26.25(t) 1.370(broad m;

1H) 1.000(broad m;

1H).sup.b 19c 10.92(q) 0.823(t;

7.4;

3H) 19d 19.82(q) 1.003(d;

6.8;

3H) 20a 30.09(q) 3.012(s;

3H) 22 21 171.39(s) 22 54.20(d) 4.761(dd;

6.5, 8.8;

1H) 18,18ab, 20,19 22a 31.42(d) 1.983(sextet;

6.7;

1H) 22b 18.18(q) 0.941(d;

6.8;

3H) 22c 16.09(q) 0.977(d;

6.8;

3H) 23 6.861(d;

8.9;

1H) 25bc 24 172.44(s) 25 76.77(d) 2.454(d;

6.9;

1H) 23 25bc 49.92(q)×

2 2.262(s;

6H) 23 26 28.08(d) 2.073(sextet;

6.7;

1H) 27 20.24(q) 0.964(d;

6.8;

3H) 28 18.18(q) 0.902(d;

6.8;

3H) ______________________________________ .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm). .sup.b Overlapping signal. .sup.c Signal assigned from NOE data.

12. A method according to claim 5 in which said neoplastic disease is murine melanoma.

13. A method according to claim 12 in which said small but effective amount comprises from about 1.44 up to about 11.1 μ

g per kilogram of host body weight.

14. A method according to claim 5 in which said neoplastic disease is human melanoma.

15. A method according to claim 14 in which said small but effective amount comprises from about 3.25 up to about 26 μ

g per kilogram of host body weight.

16. A method according to claim 11 in which said substance is administered intravenously, at a dosage level of from 0.1 up to about 20 mg per kilogram of host body weight.

17. A method according to claim 11 in which said substance is administered subcutaneously, at a dosage level of from 1 up to about 50 mg per kilogram of host body weight.

18. A method according to claim 11 in which said substance is administered orally, at a dosage level of from 5 up to about 100 mg per kilogram of host body weight.