Pharmaceutical compositions of conjugated estrogens and methods of analyzing mixtures containing estrogenic compounds

• US 6,855,703 B1
• Filed: 03/10/2000
• Issued: 02/15/2005
• Est. Priority Date: 03/10/2000
• Status: Expired due to Fees

• Current Assignee: Duramed Pharmaceuticals Incorporated (Teva Pharmaceutical Industries Limited)
• Original Assignee: Endeavor Pharmaceuticals, Inc. (Teva Pharmaceutical Industries Limited)
• Inventors: Schlipp, Katherin M., Shirazi, Dean G., Leonard, Thomas W., Hill, Edward N., Whittle, Robert R., Sancilio, Frederick D.
• Primary Examiner(s): Badio, Barbara P.
• Application Number:
  US09/524,132
• Time in Patent Office: 1,803 Days
• Field of Search: 514/170
• US Class Current: 514/170
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/565 not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
• CPC Class Codes: A61K 31/566 having an oxo group in position 17, e.g. estrone
• CPC Class Codes: A61K 31/568 substituted in positions 10 and 13 by a chain having at least one carbon atom, e.g. androstanes, e.g. testosterone
• CPC Class Codes: A61P 15/02 for disorders of the vagina
• CPC Class Codes: A61P 15/12 for climacteric disorders
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system

Abstract

A composition of matter is provided having a mixture of active estrogenic compounds. The mixture is present in chemically pure form. The mixture includes salts of conjugated estrone, conjugated equilin, conjugated Δ^8,9-dehydroestrone, conjugated 17α-estradiol, conjugated 17α-dihydroequilin, conjugated 17β-dihydroequilin, conjugated 17β-estradiol, conjugated equilenin, conjugated 17α-dihydroequilenin, and conjugated 17β-dihydroequilenin. The mixture also contains the same essential estrogenic compounds present in naturally derived equine conjugated estrogens. Drug products including the composition of matter are also provided, as are methods of using these drug products to treat mammals in need of treatment. Methods of analyzing mixtures containing conjugated estrogens are also provided.

Claims

1. A composition of matter comprising:

a mixture of estrogenic compounds, wherein said mixture comprises salts of conjugated estrone, conjugated equilin, conjugated Δ

^8,9-dehyroestrone, conjugated 17α

-estradiol, conjugated 17α

-dihydroequilin, conjugated 17β

-dihydroequilin, conjugated 17β

-estradiol, conjugated equilenin, conjugated 17α

-dihydroequilin, and conjugated 17β

-dihydroequilenin, and wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens;

wherein said composition of matter is present in a chemically pure form.

2. The composition according to claim 1, wherein the salts are sodium salts.

3. The composition according to claim 1, wherein the conjugates are sulfates.

4. The composition according to claim 1, wherein the mixture further comprises at least one compound selected from the group consisting of equilenin, 17β

-dihydroequilenin, 17β

-dihydroequilin, 17α

-dihydroequilenin, 17β

-estradiol, 17α

-dihydroequilin, 17α

-estradiol, equilin, Δ

^8,9 ehydroestrone, estrone, 17α

-Δ

^8,9-dehydroestradiol, 17β

-Δ

^8,9-dehydroestradiol, 6-OH 17α

-dihydroequilenin, 6-OH equilenin, and the salts of 17α

Δ

^8,9-dehydroestradiol sulfate, 17β

-A^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate, 6-OH 17β

-dihydroequilenin sulfate, and 6-OH equilenin sulfate.

5. The composition according to claim 4, wherein the salts of 17α

-Δ

^8,9-dehydroestradiol sulfate, 17β

-Δ

^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate and 6-OH equilenin sulfate are sodium salts.

6. The composition according to claim 1, wherein the mixture comprises:

from about 40 to about 75 percent of the salt of conjugated estrone;

from about 15 to about 40 percent of the salt of conjugated equilin;

from about 2 to about 10 percent of the salt of conjugated Δ

^8,9-dehydroestrone;

from about 2 to about 10 percent of the salt of conjugated 17α

-estradiol;

from about 10 to about 20 percent of the salt of conjugated 17α

-dihydroequilin; and

from about 0.5 to about 5 percent of the salt of conjugated 17β

-dihydroequilin.

7. The composition according to claim 1, wherein the composition further comprises at least one additional pharmaceutically active ingredient.

8. The composition according to claim 7, wherein the at least one additional pharmaceutically active ingredient is selected from the group consisting of androgens, progestins, calcium salts, and vitamin D.

21. A method of treating mammals in need of treatment for conditions treatable by administration of estrogenic compounds, said method comprising administering an effective amount of a composition of matter according to claim 1.

22. The method according to claim 21, further comprising administering the effective amount of the composition of matter in a continuous and uninterrupted manner.

23. The method according to claim 21, wherein said mammal is in need of treatment for vasomotor symptoms.

24. The method according to claim 21, wherein said mammal is in need of treatment of atrophic vaginitis.

25. The method according to claim 22, wherein said mammal is in need of treatment for osteoporosis.

9. A composition of matter comprising:

a mixture of estrogenic compounds, wherein at least one of said estrogenic compounds is a synthetic estrogenic compound, and wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens;

wherein said composition of matter is present in a chemically pure form.

12. The composition according to claim 9, wherein the mixture comprises:

from about 40 to about 75 percent of the salt of conjugated estrone;

from about 15 to, about 40 percent of the salt of conjugated equilin;

from about 2 to about 10 percent of the salt of conjugated Δ

^8,9-dehydroestrone;

from about 2 to about 10 percent of the salt of conjugated 17α

-estradiol;

from about 10 to about 20 percent of the salt of conjugated 17α

-dihydroequilin; and

from about 0.5 to about 5 percent of the salt of conjugated 17β

-dihydroequilin;

wherein the sum of the percentages of the salt of conjugated estrone and the salt of conjugated equilin is between about 70 and about 95 percent of the labeled content, and wherein the ratio of the percentage of the salt of conjugated equilin to the percentage of the salt of conjugated estrone is between about 0.25 and about 0.75.

10. The composition according to claim wherein the mixture further comprises at least one compound selected from the group consisting of equilenin, 17β

-dihydroequilenin, 17β

-dihydroequilin, 17α

-dihydroequilenin, 17β

-estradiol, 17α

-dihydroequilin, 17α

-estradiol, equilin, Δ

^8,9-dehydroestrone, estrone, 17α

-Δ

^8,9-dehydroestradiol, 17β

-Δ

^8,9-dehydroestradiol, 6-OH 17α

-dihydroequilenin, 6-OH equilenin, and the salts of 17α

Δ

^8,9-dehydroestradiol sulfate, 17β

-Δ

^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate, 6-OH 17β

-dihydroequilenin sulfate, and 6-OH equilenin sulfate.

11. The composition according to claim 10, wherein the salts of 17α

-Δ

^8,9-dehydroestradiol sulfate, 17β

-Δ

^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate and 6-OH equilenin sulfate are sodium salts.

13. A composition of matter comprising:

a mixture of estrogenic compounds, wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens;

wherein said composition of matter is present in a chemically pure form.

14. The composition according to claim 13, wherein the mixture further comprises at least one compound selected from the group consisting of equilenin, 17β

-dihydroequilenin, 17 β

-dihydroequilin, 17α

-dihydroequilenin, 17 β

-estradiol, 17α

-dihydroequilin, 17α

-estradiol, equilin, Δ

^8,9-dehydroestrone, estrone, 17α

-Δ

^8,9-dehydroestradiol, 17β

-Δ

^8,9-dehydroestradiol, 6-OH 17α

-dihydroequilenin, 6-OH equilenin, and the salts of 17α

Δ

^8,9-dehydroestradiol sulfate, 17β

-Δ

^8,9 -dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate, 6-OH 17β

-dihydroequilenin sulfate, and 6-OH equilenin sulfate.

15. The composition according to claim 14, wherein the salts of 17α

-Δ

^8,9-dehydroestradiol sulfate, 17β

-A^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate and 6-OH equilenin sulfate are sodium salts.

16. The composition according to claim 13, wherein the mixture comprises:

from about 40 to about 75 percent of the salt of conjugated estrone;

from about 15 to about 40 percent of the salt of conjugated equilin;

from about 2 to about 10 percent of the salt of conjugated Δ

^8,9-dehydroestrone;

from about 2 to about 10 percent of the salt of conjugated 17α

-estradiol;

from about 10 to about 20 percent of the salt of conjugated 17α

-dihydroequilin; and

from about 0.5 to about 5 percent of the salt of conjugated 17β

-dihydroequilin;

wherein the sum of the percentages of the salt of conjugated estrone and the salt of conjugated equilin is between about 70 and about 95 percent of the labeled content, and wherein the ratio of the percentage of the salt of conjugated equilin to the percentage of the salt of conjugated estrone is between about 0.25 and about 0.75.

17. A composition of matter comprising:

a mixture of estrogenic compounds, wherein said mixture comprises salts of conjugated estrone, conjugated equilin, conjugated Δ

^8,9-dehydroestrone, conjugated 17α

-estradiol, conjugated 17α

-dihydroequilin, conjugated 17β

-dihydroequilin, conjugated 17β

-estradiol, conjugated equilenin, conjugated 17α

-dihydroequilenin, and conjugated 17β

-dihydroequilenin, and wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens; and

at least one additional pharmaceutically active ingredient selected from the group consisting of androgens, progestins, calcium salts, and vitamin D;

wherein said composition of matter is present in a chemically pure form.

18. The composition according to claim 17, wherein the mixture further comprises at least one compound selected from the group consisting of equilenin, 17β

-dihydroequilenin, 17β

-dihydroequilin, 17α

-dihydroequilenin, 17β

-estradiol, 17α

-dihydroequilin, 17α

-estradiol, equilin, Δ

^8,9-dehydroestrone, estrone, 17α

-Δ

^8,9-dehydroestradiol, 17β

Δ

^8,9-dehydroestradiol, 6-OH 17α

-dihydroequilenin, 6-OH equilenin, and the salts of 17α

Δ

^8,9-dehydroestradiol sulfate, 17β

-Δ

^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate, 6-OH 17β

-dihydroequilenin sulfate, and 6-OH equilenin sulfate.

19. The composition according to claim 18, wherein the salts of 17α

-Δ

^8,9-dehydroestradiol sulfate, 17 β

-Δ

^8,9-dehydroestradiol sulfate, 6-OH 17α

-dihydroequilenin sulfate, 6-OH 17β

-dihydroequilenin sulfate, and 6-OH equilenin sulfate are sodium salts.

20. The composition according to claim 17, wherein the mixture comprises:

from about 40 to about 75 percent of the salt of conjugated estrone;

from about 15 to about 40 percent of the salt of conjugated equilin;

from about 2 to about 10 percent of the salt of conjugated Δ

^8,9-dehydroestrone;

from about 2 to about 10 percent of the salt of conjugated 17α

-estradiol;

from about 10 to about 20 percent of the salt of conjugated 17α

-dihydroequilin; and

from about 0.5 to about 5 percent of the salt of conjugated 17β

-dihydroequilin;

wherein the sum of the percentages of the salt of conjugated estrone and the salt of conjugated equilin is between about 70 and about 95 percent of the labeled content of conjugated estrogens, and wherein the ratio of the percentage of the salt of conjugated equilin to the percentage of the salt of conjugated estrone is between about 0.25 and about 0.75.

26. A method for analyzing conjugated estrogen constituents comprising the steps of:

preparing a solution containing conjugated estrogens, said solution comprising;

a mixture comprising estrogenic compounds to be analyzed; and

a mobile phase comprising;

an organic portion comprising between about 0.1% and about 30% (by volume organic portion)-protic solvent and between about 70% and about 100% (by volume organic portion) polar aprotic solvent; and

an aqueous portion;

and analyzing the conjugated estrogens solution utilizing a HPLC system.

27. The method according to claim 26, wherein the mixture comprising estrogen compounds comprises naturally derived equine conjugated estrogens.

28. The method according to claim 26, wherein the protic solvent comprises a lower alkyl alcohol.

29. The method according to claim 28, wherein the lower alkyl alcohol is methanol.

30. The method according to claim 26, wherein the polar aprotic solvent comprises a lower alkyl nitrile.

31. The method according to claim 30, herein the lower alkyl nitrile is acetonitrile.

32. The method according to claim 26, wherein the mobile phase comprises between about 65 and about 80 percent aqueous portion and between about 20 and about 35 percent organic portion.

33. The method according to claim 26, wherein the aqueous portion has a pH of between about 2.5 to about 3.5.

34. The method according to claim 26, wherein the organic portion comprises an ion-pairing agent.

35. The method according to claim 34 wherein the ion-pairing agent has a concentration between about 0.5 to about 2 mM.

36. The method according to claim 34, wherein the ion-pairing agent is tert-butyl ammonium hydroxide.

37. The method according to claim 36, wherein the aqueous portion comprises an ion-pairing agent.

38. The method according to claim 37, wherein the ion-pairing agent has a concentration between about 0.5 to about 2 mM.

39. The method according to claim 37, wherein the ion-pairing agent is tert-butyl ammonium hydroxide.

40. The method according to claim 26, wherein the step of analyzing further comprises the step of fraction collecting at least one of the separated constituents of the conjugated estrogens substance.

41. The method according to claim 40, wherein the step of fraction collecting utilizes a multi-channel fraction collector.

42. The method according to claim 41, wherein the HPLC system comprises a column, and wherein the analyzing step occurs at a column temperature between about 15 and about 35°

C.

43. A composition of matter comprising:

a mixture of estrogenic compounds, wherein said mixture comprises salts of conjugated estrone, conjugated equilin, conjugated Δ

^8,9-dehydroestrone, conjugated 17α

-estradiol, conjugated 17α

-dihydroequilin, conjugated 17β

-dihydroequilin, conjugated 17β

-estradiol, conjugated equilenin, conjugated 17α

-dihydroequilenin, and conjugated 17β

-dihydroequilenin, and wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens;

wherein said composition of matter is substantially devoid of a compound selected from the group consisting of indican, sulfated benzyl alcohol, hippuric acid, benzoic acid, and creatinine.

44. A composition of matter comprising:

a mixture of estrogenic compounds, wherein said mixture comprises salts of conjugated estrone, conjugated equilin, conjugated Δ

^8,9-dehydroestrone, conjugated 17α

-estradiol, conjugated 17α

-dihydroequilin, conjugated 17β

-dihydroequilin, conjugated 17β

-estradiol, conjugated equilenin, conjugated 17α

-dihydroequilenin, and conjugated 17β

-dihydroequilenin, and wherein said mixture comprises the same essential estrogenic compounds present in naturally derived equine conjugated estrogens;

wherein said composition of matter is substantially devoid of indican, sulfated benzyl alcohol, hippuric acid, benzoic acid, and creatinine.