Preparation of preliposomal annamycin lyophilizate

Abstract

A method for preparing lyophilized anamycin liposome precursors, a composition prepared by the method, and the use of the composition prepared therefrom in the treatment of cancer are provided.

Description

[Technical Field] This invention relates to a method for preparing anamycin, and more specifically, to a method for preparing lyophilized anamycin. This invention also relates to a composition obtained by this method, and its use in the manufacture of medicaments for treating cancer. [Previous Technology] Annamycin is a cancer chemotherapeutic agent belonging to the anthracnose family. Due to its physical and pharmacological properties, dosage forms of annamycin formulations, in which the drug is formulated as liposomes, have been described (see US 7,238,366). Clinical trials of liposomal annamycin have been described in adult patients with relapsed / refractory acute lymphoblastic leukemia (M. Wetzler et al., Clinical Lymphoma, Myeloma and Leukemia, 13(4), 430-434, August 2013) and in the treatment of doxorubicin-resistant breast cancer (DJ Booser et al., Cancer Chemother. Pharmacol. 50; 6-8, 2002). In drug delivery systems, it is desirable to produce drug formulations with consistent purity and quality to ensure the correct dosage is administered, thereby providing the most effective treatment and avoiding undesirable side effects caused by impurities. Furthermore, poorly maintained or improperly formulated drug materials may decompose before administration, rendering them ineffective and wasting expensive active pharmaceutical ingredients. Regulatory standards must also ensure consistent levels of drug purity and dosage according to the procedures specified on the label. Anamycin presents a particularly challenging formulation due to its inherent physical properties and the inherent instability of both the parent drug and the liposomal formulation under certain conditions. Liposome formulations of anamycin often degrade and form anamycin crystals, and / or form large liposomes or undergo phase separation, rendering the formulation unsuitable for administration. Therefore, finding a consistent method for preparing anamycin liposomal preforms that ensures purity and avoids anamycin degradation would be advantageous. [Summary of the Invention] A method for preparing lyophilized anamycin is provided, the method comprising the following steps: preparing a solution comprising one or more lipids, one or more nonionic surfactants and one or more solvents and having a pH of 4.8-5.9; adding anamycin solution to the lipid solution, the anamycin solution comprising about 8-12 wt.% anamycin in DMSO to provide a lipid-containing anamycin solution; aseptically filtering the lipid-containing anamycin solution; and lyophilizing the lipid-containing anamycin solution to provide anamycin liposome precursor lyophilized product. A method for producing lyophilized anamycin is provided, the method comprising the steps of: preparing a first solution comprising water and terbutanol, wherein the ratio of water to terbutanol is from about 8:2 to about 9:1; adding one or more lipids and one or more nonionic surfactants to the first solution to form a second solution; adjusting the pH of the second solution to a pH of 4.8–5.9; adding a third solution to the second solution, the third solution comprising about 8–12 wt.% anamycin in DMSO to provide a lipid-containing anamycin solution; aseptically filtering the lipid-containing anamycin solution; and lyophilizing the lipid-containing anamycin solution to provide anamycin liposome precursor lyophilized product. A method for producing ananamycin liposome precursor lyophilized product is also provided, the method comprising the following steps: adding DMPC, DMPG and polysorbate 20 to a mixture of preheated water and tert-butanol to prepare a first solution, wherein the preheating temperature is from about 35°C to about 42°C, and wherein the ratio of water to tert-butanol is from about 8:2 to about 9:1; adjusting the pH of the first solution with a pharmaceutically acceptable acid to maintain a constant pH of the solution for at least 15 minutes; adding a second solution to the first solution, the second solution containing about 8-12 wt% ananamycin in DMSO to provide a lipid-containing ananamycin solution; aseptically filtering the lipid-containing ananamycin solution; and lyophilizing the lipid-containing ananamycin solution in separate aliquots to provide liposome precursor lyophilized product. Also provided are lyophilized anamycin liposome precursors prepared by any of the methods described herein. It also provides a method for treating cancer, which includes administering an effective dose of liposomal anamycin to a patient in need, the liposomal anamycin being prepared using an anamycin liposomal precursor lyophilized product prepared by any of the methods described herein. The use of lyophilized anamycin liposome precursors prepared according to any of the methods described herein in the manufacture of medicaments for the treatment of cancer is also provided. Also provided are lyophilized anamycin liposome precursors prepared according to any of the methods described herein for use in the treatment of cancer.

Implementation Method

Claims

1. A method for preparing lyophilized anamycin, the method comprising the following steps: A solution comprising one or more lipids, one or more nonionic surfactants and one or more solvents and having a pH of 4.8-5.9 is prepared; wherein the one or more lipids are DMPG and the pH is adjusted to 4.8-5.9, and wherein the one or more nonionic surfactants are polyoxyethylene sorbitan surfactants. An anamycin solution containing 8-12 wt.% anamycin in DMSO is added to a lipid solution to provide a lipid-containing anamycin solution; the lipid-containing anamycin solution is aseptically filtered; and the lipid-containing anamycin solution is lyophilized to provide an anamycin liposome precursor lyophilized product, wherein the anamycin in the resulting lyophilized product is at least 95% pure.

2. A method for preparing lyophilized anamycin, the method comprising the steps of: preparing a first solution comprising water and terbutanol, wherein the ratio of water to terbutanol is from 8:2 to 9:1; adding one or more lipids and one or more nonionic surfactants to the first solution to form a second solution; wherein the one or more lipids comprise DMPG, and wherein the one or more nonionic surfactants comprise polyoxyethylene sorbitan surfactants; adjusting the pH of the second solution to a pH of 4.8-5.9; adding a third solution to the second solution, the third solution comprising 8-12 wt.% anamycin in DMSO to provide a lipid-containing anamycin solution; aseptically filtering the lipid-containing anamycin solution; and lyophilizing the lipid-containing anamycin solution to provide anamycin liposome precursor lyophilized product, wherein the anamycin in the resulting lyophilized product is at least 95% pure.

3. The method as described in claim 2, wherein the second solution comprises DMPC, DMPG and polyoxyethylene sorbitan surfactant.

4. The method as described in claim 2 or 3, wherein the second solution comprises DMPC, DMPG and polyoxyethylene sorbitan monolaurate.

5. The method as described in claim 2 or 3, wherein the pH of the second solution is adjusted to 5.3 ± 0.2 with a pharmaceutically acceptable acid selected from the group consisting of HCl and H2SO4.

6. The method as described in claim 2 or 3, wherein the third solution contains 10 wt.% anamycin in DMSO.

7. The method as described in claim 2 or 3, wherein each lipid and each surfactant are added separately.

8. The method as described in claim 4, wherein DMPC, DMPG and polyoxyethylene sorbitan monolaurate are added sequentially to the first solution.

9. The method as described in claim 2 or 3, wherein the anamycin liposome precursor lyophilized product comprises: 1.8-2.2 wt.% anamycin, 3.0-3.4 wt.% polysorbate 20, and 94.4-95.2 wt.% lipids selected from DMPC and DMPG.

10. The method as described in claim 9, wherein the anamycin in the resulting lyophilized product is at least 98% pure.

11. The method as described in claim 10, wherein the anamycin in the resulting lyophilized product is at least 99% pure.

12. The method as described in claim 4, further comprising: After initially adjusting the pH of the second solution to pH 5.3 ± 0.2, allow the solution to equilibrate for at least 15 minutes; Measure the pH and adjust it with acid if the pH is above 5.3; Allow the solution to equilibrate for at least 15 minutes; And repeat the following steps: measure pH, adjust pH, and equilibrate the solution until the pH of the solution is maintained at pH 5.3 ± 0.

2.

13. The method as described in claim 12, wherein the acid used to adjust the pH is selected from the group consisting of HCl and H2SO4.

14. The method as described in claim 12 or 13, wherein after each addition of acid, the second solution is allowed to equilibrate for at least 30 minutes.

15. The method as described in claim 2 or 3, wherein each solution is maintained at a temperature from 38°C to 42°C until lyophilization begins.

16. A method for producing ananamycin liposome precursor lyophilized product, the method comprising the steps of: adding DMPC, DMPG and polysorbate 20 to a preheated mixture of water and tert-butanol to prepare a first solution, wherein the preheating temperature is from 35°C to 42°C and wherein the ratio of water to tert-butanol is from 8:2 to 9:1; adjusting the pH of the first solution to 4.8-5.9 with one or more pharmaceutically acceptable acids to maintain a constant pH of the solution for 15-90 minutes; adding a second solution to the first solution, the second solution containing 8-12 wt.% ananamycin in DMSO to provide a lipid-containing ananamycin solution; aseptically filtering the lipid-containing ananamycin solution; and lyophilizing the lipid-containing solution in separate aliquots to provide ananamycin liposome precursor lyophilized product, wherein the ananamycin in the resulting lyophilized product is at least 95% pure.

17. The method as described in claim 16, wherein the second solution contains 10 wt.% anamycin in DMSO and the pH is 5.3 ± 0.

2.

18. A composition of an anamycin liposome precursor lyophilized product prepared by any one of claims 1 to 17, the composition comprising: anamycin, one or more lipids, and one or more nonionic surfactants.

19. The composition as claimed in claim 18, comprising: 1.8-2.2 wt.% anamycin, 3.0-3.4 wt.% polysorbate 20, and 94.4-95.2 wt.% lipids selected from DMPC and DMPG.

20. The composition as described in claim 19, wherein the DMPC is 65.3-67.3 wt.% and the DMPG is 27.1-29.9 wt.%.

21. Use of liposomal anamycin in the preparation of a medicament for treating cancer, wherein the liposomal anamycin is prepared by reconstructing an anamycin liposomal precursor lyophilized product obtained by any one of claims 1-17.

22. The use as described in claim 21, wherein the cancer is selected from the group consisting of leukemia and lymphoma.