Stable formulations of 5,10-methylene-(6R)-tetrahydrofolic acid

Abstract

The invention is directed towards stable formulations of 5,10-methylene-(6R)-tetrahydrofolic acid and its hemisulfate salt as well as pharmaceutical compositions and uses thereof in therapy, preferably chemotherapy.

Claims

1. A method for treating breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesothelioma cancer, stomach cancer, bowel cancer, or lung cancer, comprising administering an effective amount of a pharmaceutical composition to a patient in need thereof, which composition comprises: A) a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, B) a lyophilisate comprising the hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid of A) or made therefrom, C) a reconstituted solution from the lyophilisate of B), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle, D) a lyophilisate comprising the sulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom without the presence of citrate, E) a lyophilisate comprising 5,10-methylene-(6R)-tetrahydrofolic acid without the presence of citrate, or F) a reconstituted solution from the lyophilisate of D) or E), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

2. A method for methotrexate therapy, comprising administering methotrexate to a subject in need thereof, and additionally administering A) a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, B) a lyophilisate comprising the hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid of A) or made therefrom, C) a reconstituted solution from the lyophilisate of B), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle, D) a lyophilisate comprising the sulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom without the presence of citrate, E) a lyophilisate comprising 5,10-methylene-(6R)-tetrahydrofolic acid without the presence of citrate, or F) a reconstituted solution from the lyophilisate of D) or E), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

3. A method for treating breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesothelioma cancer, stomach cancer, bowel cancer, or lung cancer, comprising administering an effective amount of a pharmaceutical composition to a patient in need thereof, which composition comprises a lyophilized formulation A, B, C, D or E, or a method for methotrexate therapy, comprising administering methotrexate to a subject in need thereof, and additionally administering a pharmaceutical composition comprising a lyophilized formulation A, B, C, D or E, which A) is prepared from a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, B) comprises 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate resulting from a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, C) comprises about two moles of 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate to one mole of sulfate ions, D) comprises about 1 mole of 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate to one mole of sulfate ions without the presence of citrate, or E) comprises 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate having a chemical purity of greater than 99%, and sulfate with the presence of citrate or without the presence of citrate; or which is a stable pharmaceutical composition X or Y comprising X) a lyophilised hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, which has such stability that the composition maintains greater than or equal to 95% of purity of the 5,10-methylene-(6R)-tetrahydrofolic acid for at least 24 months at +25 C./60% relative humidity; or for at least 12 months at +40 C./75% relative humidity, or for at least 24 months at +5 C., or for at least 24 months at 20 C.; or Y) a reconstituted lyophilised hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, which has such stability that the composition maintains greater than or equal to 95% of purity of the 5,10-methylene-(6R)-tetrahydrofolic acid for at least 200 minutes at 2-8 C., or for at least 2 hours at room temperature.

4. A method according to claim 1, wherein the hemisulfate salt has been dissolved in water or in a liquid pharmaceutically acceptable vehicle, wherein the resultant solution has a pH of between 5 and 10.5.

5. A method according to claim 1, wherein the hemisulfate salt is in crystalline form, wherein the hemisulfate salt has one or more X-ray pattern peak positions at an angle of diffraction 2 theta of 4.7, 17.9, and 23.3 expressed in 20.2 2 CuK radiation, reflection; or wherein the hemisulfate salt has a FT-Raman spectrum containing one or more peaks at wavenumbers, expressed in 2 cm.sup.1, of 1672, 1656, 1603, 1553, 1474, 1301, 637, 624 and 363; or wherein the hemisulfate salt has a FT-Raman spectrum substantially in accordance with FIG. 1; or wherein the hemisulfate salt has an X-ray powder diffraction pattern substantially in accordance with FIG. 2(a); or wherein the hemisulfate salt has an X-ray powder diffraction pattern substantially in accordance with FIG. 2(b).

6. A method according to claim 1, wherein the hemisulfate salt has a stereoisomeric purity of greater than 99%; or has a chemical purity of greater than 99%.

7. A method according to claim 1, wherein in the lyophilisate the molar ratio between the (6R)-5,10-CH.sub.2-THF and sulfuric acid moieties is from about 1:1 to about 2:1.

8. A method according to claim 1, wherein the hemisulfate salt is in a pharmaceutical composition that also comprises a buffer, which buffer is citrate, phosphate, acetate, TRIS, N-tris(hydroxymethyl) methyl-2-aminoethanesulfonic acid (TES), 3-(N-morpholino) propanesulfonic acid (MOPS), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), MES, MOPSO, HEPES, succinate, carbonate, ammonium, mono-, di- or tri-alkylammonium, mono-, di- or tri-hydroxylalkylammonium, maleate, glutamate, borate, lactate or a combination thereof.

9. A method according to claim 3, wherein colorectal cancer is treated, which is metastatic colorectal cancer.

10. A method according to claim 3, wherein lung cancer is treated, which is adenocarcinoma.

11. A method according to claim 1, wherein colorectal cancer is treated, which is metastatic colorectal cancer.

12. A method according to claim 1, wherein lung cancer is treated, which is adenocarcinoma.

13. A method according to claim 1, wherein colon cancer, rectal cancer, colorectal cancer or bowel cancer is treated.

14. A method according to claim 1, wherein gastric cancer or stomach cancer is treated.

15. A method according to claim 1, wherein breast cancer or ovarian cancer is treated.

16. A method according to claim 1, wherein gall bladder cancer, bile duct cancer, liver cancer or pancreatic cancer is treated.

17. A method according to claim 1, wherein esophageal cancer is treated.

18. A method according to claim 1, wherein osteosarcoma is treated.

19. A method according to claim 1, wherein head and neck cancer is treated.

20. A method according to claim 1, wherein mesothelioma cancer is treated.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: Raman spectrum of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1), recorded using a nominal laser power level of 300 mW and 64 scans.

(2) FIG. 2 (a): X-Ray Powder Diffractogram of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1) recorded in the reflection mode;

(3) FIG. 2 (b): X-Ray Powder Diffractogram of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1) recorded in the transmission mode;

(4) FIG. 2 (c): Comparison of X-ray diffraction pattern of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1) recorded in transmission mode (upper curve A) with a recording of the same compound recorded in reflection mode (lower curve B);

(5) FIG. 2 (d): Comparison of X-ray diffraction pattern of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1) (upper curve A) with an X-ray diffraction pattern of the sulfate salt of (6R)-5,10-CH.sub.2-THF (lower curve B) recorded in the transmission mode.

(6) FIG. 3: TG-FTIR thermogram of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1). A indicates a change in mass of 0.5% (due to loss of water) and B indicates a change in mass of 14.53% (due to decomposition)

(7) FIG. 4: DSC thermogram of the hemisulfate salt of (6R)-5,10-CH.sub.2-THF (Type 1; first scan: solid line; second scan (after quench cooling): dashed line).

(8) FIG. 5: Shows HPLC purity results of the lyophilised hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid through 24 months of testing at all tested conditions.

(9) FIG. 6: Illustrates stability data at 20 C. of the sulfate salt in comparison to data generated by measurements on three batches of the hemisulfate salt provided.

(10) FIG. 7: Illustrates stability marker results graphically at 20 C. of the sulfate salt in comparison to data generated by measurements on three batches of the hemisulfate salt.

(11) FIG. 8: Illustrates the showing of improved stability data at various temperatures by measurements on three batches of the hemisulfate salt in comparison to the sulfate salt.

(12) FIG. 9: Illustrates stability marker results graphically for the showing of improved stability at various temperatures by measurements on three batches of the hemisulfate salt in comparison to the sulfate salt.

(13) FIG. 10: Illustrates tabulated data for Example 4-1.

(14) In sum, disclosed are the following aspects:

(15) Aspect 1: A hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid.

(16) Aspect 2: The hemisulfate salt according to aspect 1, which has been dissolved in water or in a liquid pharmaceutically acceptable vehicle, wherein the resultant solution has a pH of between 5 and 10.5.

(17) Aspect 3: The hemisulfate salt of aspect 1, which is anhydrous; or which is in crystalline form; or which is in amorphous form.

(18) Aspect 4: A lyophilisate comprising the hemisulfate salt of aspect 2 or made therefrom.

(19) Aspect 5: The hemisulfate salt of aspect 3, which is in crystalline form, wherein the hemisulfate salt has one or more X-ray pattern peak positions at an angle of diffraction 2 theta of 4.7, 17.9, and 23.3 expressed in 20.2 2 CuK radiation, reflection; or wherein the hemisulfate salt has a FT-Raman spectrum containing one or more peaks at wavenumbers, expressed in 2 cm.sup.1, of 1672, 1656, 1603, 1553, 1474, 1301, 637, 624 and 363; or wherein the hemisulfate salt has a FT-Raman spectrum substantially in accordance with FIG. 1; or wherein the hemisulfate salt has an X-ray powder diffraction pattern substantially in accordance with FIG. 2(a); or wherein the hemisulfate salt has an X-ray powder diffraction pattern substantially in accordance with FIG. 2(b).

(20) Aspect 6: The hemisulfate salt of aspect 1, having a stereoisomeric purity of greater than 99%; preferably having a stereoisomeric purity of greater than 99.5%; or having a chemical purity of greater than 99%; preferably having a chemical purity of greater than 99.5%.

(21) Aspect 7: A pharmaceutical composition comprising a hemisulfate salt of aspect 1 and a pharmaceutically acceptable carrier.

(22) Aspect 8: A pharmaceutical composition of aspect 7 further comprising a buffer, which buffer is preferably citrate, phosphate, acetate, TRIS, N-tris(hydroxymethyl) methyl-2-aminoethanesulfonic acid (TES), 3-(N-morpholino) propanesulfonic acid (MOPS), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), MES, MOPSO, HEPES, succinate, carbonate, ammonium, mono-, di- or tri-alkylammonium, mono-, di- or tri-hydroxylalkylammonium, maleate, glutamate, borate, lactate or a combination thereof, and more preferably the buffer is citrate.

(23) Aspect 9: The lyophilisate of aspect 4, wherein the molar ratio between the (6R)-5,10-CH.sub.2-THF and sulfuric acid moieties is from about 1:1 to about 2:1.

(24) Aspect 10: A process for preparing a lyophilized product comprising the step of lyophilizing a pharmaceutical composition comprising a hemisulfate salt of aspect 1 and a pharmaceutically acceptable carrier.

(25) Aspect 11: A process for preparing a liquid pharmaceutical composition from the lyophilisate of aspect 4 comprising dissolving the lyophilized pharmaceutical composition in water and/or a liquid pharmaceutically acceptable vehicle.

(26) Aspect 12: A method for treating breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma), comprising administering an effective amount of a pharmaceutical composition to a patient in need thereof, which composition comprises:

(27) A) a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid according to aspect 1,

(28) B) a lyophilisate comprising the hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom,

(29) C) a reconstituted solution from the lyophilisate of B), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle;

(30) and which method preferably includes also administering to said patient an effective amount of a further pharmaceutically active compound, e.g., a cytotoxic compound, for example, 5-fluorouracil, pemetrexed, high-dose methotrexate, oxaliplatin, avastin, etc., which further active compound is suitable for the treatment of said breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma). One could use the pharmaceutical composition recited in this method for the treatment of indications for which Levoleucovorin (or Leucovorin) is used. For example, the 5-FU based chemotherapy is based on the inhibition of TS because of the very special, unnatural, ternary complex.

(31) Aspect 13: A product, which is i) a lyophilisate comprising the sulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom without the presence of citrate, wherein preferably the lyophilisate comprises the 5,10-methylene-(6R)-tetrahydrofolic acid and sulfate and optionally sodium ions without the presence of citrate, ii) a lyophilisate comprising 5,10-methylene-(6R)-tetrahydrofolic acid without the presence of citrate, iii) a reconstituted solution from the lyophilisate of i) or ii), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

(32) Aspect 14: A method for treating breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma), comprising administering an effective amount of a pharmaceutical composition to a patient in need thereof, which composition comprises a product according to aspect 13; and which method preferably includes also administering to said patient an effective amount of a further pharmaceutically active compound, e.g., a cytotoxic compound, for example, 5-fluorouracil, pemetrexed, high-dose methotrexate, oxaliplatin, avastin, etc., which further active compound is suitable for the treatment of said breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma). One could use the pharmaceutical composition recited in this method for the treatment of indications for which Levoleucovorin (or Leucovorin) is used. For example, the 5-FU based chemotherapy is based on the inhibition of TS because of the very special, unnatural, ternary complex.

(33) Aspect 15: A reconstituted solution from a lyophilisate according to aspect 4, which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

(34) Aspect 16: A reconstituted solution according to aspect 15, wherein the molar ratio between the (6R)-5,10-CH.sub.2-THF and H.sub.2SO.sub.4 moieties is from about 1:1 to about 2:1, preferably about 2:1.

(35) Aspect 17: A product, which is a lyophilized formulation A, B, C, D or E, which A) is prepared from a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, B) comprises 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate resulting from a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, C) comprises about two moles of 5,10-methylene-(6M-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate to one mole of sulfate ions, D) comprises about 1 mole of 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate to one mole of sulfate ions without the presence of citrate, E) comprises 5,10-methylene-(6R)-tetrahydrofolic acid and/or 5,10-methylene-(6R)-tetrahydrofolate having a chemical purity of greater than 99%, more preferably 99.5%, or even higher, e.g., 99.6, 99.7, 99.8 or 99.9% or above, e.g., 100, and sulfate with the presence of citrate or without the presence of citrate; or
which is a stable pharmaceutical composition X or Y comprising 5,10-methylene-(6R)-tetrahydrofolic acid, which composition comprises X) a lyophilised hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, which has such stability that the composition maintains greater than or equal to 95%, preferably between 97-98%, and more preferably at about 99%, of purity of the 5,10-methylene-(6R)-tetrahydrofolic acid for at least 24 months, preferably for at least 36 months, and more preferably for at least 48 months at +25 C./60% relative humidity; or for at least 12 months at +40 C./75% relative humidity, or for at least 24 months, preferably at least 36 months, and more preferably for at least 48 months at +5 C., or for at least 24 months, preferably at least 36 months at 20 C.; or Y) a reconstituted lyophilised hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid, which has such stability that the composition maintains greater than or equal to 95%, preferably between 95-96%, and more preferably at about 97%, of purity of the 5,10-methylene-(6R)-tetrahydrofolic acid for at least 200 minutes, preferably 6 hours, and more preferably 11 or 12 hours at 2-8 C., or for at least 2 hours, and more preferably for at least 200 minutes at room temperature.

(36) Aspect 18: A method for treating breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma), comprising administering an effective amount of a pharmaceutical composition to a patient in need thereof, which composition comprises a product according to aspect 17; and which method preferably includes also administering to said patient an effective amount of a further pharmaceutically active compound, e.g., a cytotoxic compound, for example, 5-fluorouracil, pemetrexed, high-dose methotrexate, oxaliplatin, avastin, etc., which further active compound is suitable for the treatment of said breast cancer, esophageal cancer, gastric cancer, gall bladder cancer, bile duct cancer, colon cancer, rectal cancer, colorectal cancer (CRC) including metastatic CRC, osteosarcoma, liver cancer, pancreatic cancer, ovarian cancer, head and neck cancer, mesotheolioma cancer, stomach cancer, bowel cancer, or lung cancer (specifically adenocarcinoma). One could use the pharmaceutical composition recited in this method for the treatment of indications for which Levoleucovorin (or Leucovorin) is used. For example, the 5-FU based chemotherapy is based on the inhibition of TS because of the very special, unnatural, ternary complex.

(37) Aspect 19: A method for methotrexate therapy, comprising administering methotrexate to a subject in need thereof, and additionally administering A) a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid according to aspect 1, B) a lyophilisate comprising the hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom, C) a reconstituted solution from the lyophilisate of B), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

(38) Aspect 20: In a method for methotrexate therapy, wherein the improvement comprises administering A) a hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid according to aspect 1, B) a lyophilisate comprising the hemisulfate salt of 5,10-methylene-(6R)-tetrahydrofolic acid or made therefrom, C) a reconstituted solution from the lyophilisate of B), which has been reconstituted by water or a liquid pharmaceutically acceptable vehicle.

(39) Aspect 21: A method for methotrexate therapy, comprising administering methotrexate to a subject in need thereof, and additionally administering a product of aspect 13.

(40) Methotrexate therapy is typically used in the treatment of various types of cancers as well as various autoimmune diseases. Cancers for which methotrexate therapy is used include skin cancers, breast cancer, head and neck cancer, e.g., epidermoid cancers of the head and neck, lung cancer, particularly squamous cell and small cell types, leukemia, e.g., acute lymphoblastic leukemia (ALL), lymphoma, e.g., non-Hodgkin's lymphoma (NHL) and central nervous system (CNS) lymphoma, mycosis fungoides (cutaneous T-cell lymphoma), colon cancer, colorectal cancer, rectal cancer, stomach cancer, esophageal cancer, choriocarcinoma, chorioadenoma, gestational trophoblastic disease, osteosarcoma, nonleukemic meningeal cancer, soft tissue sarcoma (desmoid tumors, aggressive fibromatosis) and bladder cancer. Methotrexate therapy is also used in the treatment of psoriasis, rheumatoid arthritis, Crohn's disease, juvenile dermatomyositis, psoriatic arthritis, lupus, sarcoidosis, eczema, vasculitis, dermatomyositis/polymyositis, ectopic pregnancy, systemic lupus erythematosus, and Takayasu arteritis, and for prevention of graft-versus-host disease.

(41) The use of the compounds of the invention in methotrexate therapy can rather be considered a rescue cancer therapy. In this case, there isn't the same inhibitory ternary complex that includes, for example, the 5-FU molecule, but instead is the normal ternary complex that includes uracil instead of 5-FU, and which instead speeds up the normal formation of thymidine and DNA, and both in the normal cells and in the tumours.

(42) From the preceding description, one skilled in the art in this field can read the elements of the invention without problems, and, without departing from the basic idea and from the scope of the invention, can make modifications and additions and can thereby adapt the invention to differing needs and conditions.

(43) The entire disclosures of all the patent applications, patents and publications which are cited in this text are included jointly by reference.