High purity oritavancin and method of producing same
Drug substance preparations of oritavancin having high purity are disclosed, along with pharmaceutical compositions comprising such oritavancin drug substance preparations, and drug products or dosage forms comprising such pharmaceutical compositions.
1. A method for preparing an oritavancin drug substance preparation, comprising:
a) growing a culture of a chloroeremomycin-producing microorganism under fermentative conditions in a medium free of animal-sourced material (ASM) and under conditions promoting biosynthesis of chloroeremomycin by the culture,
b) recovering chloroeremomycin from fermentation broth produced in a),
c) preparing a solution of the chloroeremomycin of b) and a copper salt in an organic solvent, reacting the solution with 4-chloro-4′-biphenyl carboxaldehyde, and precipitating oritavancin-copper complex from the solution,
d) de-complexing copper from the oritavancin-copper complex of c), and
e) collecting oritavancin, thereby preparing an oritavancin drug substance preparation,
wherein the oritavancin drug substance preparation has a maximum impurity level of not more than 3.0% by peak area of impurity 1 and impurity 7, defined by peak A of FIG. 1 and peak G of FIG. 2 , respectively; or
wherein the oritavancin drug substance preparation has about 85% purity or greater by peak area relative to impurities 1-16, defined by peak A of FIG. 1 and peaks B-P of FIG. 2 , respectively.
2. The method of claim 1 , wherein the chloroeremomycin is recovered from fermentation broth produced in a) using a polymeric exchange resin.
3. The method of claim 1 , further comprising decolorizing the chloroeremomycin recovered in b) using a polymeric adsorbent resin, chromatographically separating the decolorized chloroeremomycin using a hydrophobic polymeric resin column, and precipitating the separated chloroeremomycin using an organic solvent.
4. The method of claim 1 , wherein the oritavancin-copper complex is precipitated from the solution in c) using acetonitrile.
5. The method of claim 1 , wherein the copper is de-complexed from the oritavancin-copper complex of d) by adding an aqueous acid and separating the de-complexed oritavancin using a polymeric hydrophobic resin, wherein the adding and separating are performed concurrently or sequentially.
6. The method of claim 5 , further comprising collecting oritavancin via:
(i) eluting oritavancin from the polymeric hydrophobic resin,
(ii) concentrating oritavancin eluted from the resin,
(iii) precipitating oritavancin from the concentrate in aqueous ethanol, and
(iv) drying the precipitated oritavancin.
7. The method of claim 1 , wherein the purity level of the oritavancin drug substance preparation is measured by HPLC.
8. The method of claim 1 , wherein the purity level of the oritavancin drug substance preparation is measured by HPLC, and wherein the HPLC measurement utilizes a C18 reverse-phase stationary phase and a gradient of mobile phase B, which is phosphoric acid/water/acetonitrile/tetrahydrofuran at a ratio of about 1/1000/1500/25 (v/v/v/v), in mobile phase A, which is phosphoric acid/water/tetrahydrofuran at a ratio of about 1/1000/10 (v/v/v).
9. The method of claim 1 , wherein the chloroeremomycin-producing microorganism is a microorganism of genus Nocardia , genus Amycolatopsis , or genus Kibdelosporangium.
10. The method of claim 1 , wherein the chloroeremomycin-producing microorganism is Kibdelosporangium aridum.
11. The method of claim 2 , wherein the polymeric exchange resin is a sulfonated macroporous copolymer of styrene and divinylbenzene.
12. The method of claim 3 , wherein the polymeric adsorbent resin or the hydrophobic polymeric resin column is a non-functionalized macroporous copolymer of styrene and divinylbenzene.
13. The method of claim 5 , wherein the polymeric hydrophobic resin is a non-functionalized macroporous copolymer of styrene and divinylbenzene.
14. The method of claim 1 , wherein the organic solvent is methanol.
15. The method of claim 3 , wherein the organic solvent is methanol.
16. The method of claim 5 , wherein the aqueous acid is formic acid and/or phosphoric acid.
17. The method of claim 6 , wherein the concentrating of the oritavancin solution is via distillation of volatile solvents under reduced pressure or ultrafiltration/diafiltration.
18. The method of claim 6 , wherein the drying of the precipitated oritavancin is via drying on a tray or in a Nutsche filter at elevated temperature and reduced pressure.
19. The method of claim 3 , wherein the method further comprises performing a concentrating step and a precipitating step between the decolorizing and chromatography.
20. The method of claim 1 , wherein the method further comprises terminating the reacting step of c) using a hydride reagent.
21. A method for preparing an oritavancin drug substance preparation, comprising:
a) growing a culture of a chloroeremomycin-producing microorganism under fermentative conditions in a medium free of animal-sourced material (ASM) and under conditions promoting biosynthesis of chloroeremomycin by the culture,
b) recovering chloroeremomycin from fermentation broth produced in a) using a polymeric exchange resin,
c) decolorizing the chloroeremomycin recovered in b) using a polymeric adsorbent resin, chromatographically separating the decolorized chloroeremomycin using a hydrophobic polymeric resin column, and precipitating the separated chloroeremomycin using an organic solvent,
d) preparing a solution of the precipitated chloroeremomycin of c) and a copper salt in an organic solvent, reacting the solution with 4-chloro-4′-biphenyl carboxaldehyde, and precipitating oritavancin-copper complex from the solution using acetonitrile,
e) de-complexing copper from the oritavancin-copper complex of d) by adding an aqueous acid and separating the de-complexed oritavancin using a polymeric hydrophobic resin, wherein the adding and separating are performed concurrently or sequentially,
f) eluting oritavancin from the polymeric hydrophobic resin of e),
g) concentrating oritavancin eluted from the resin in f),
h) precipitating oritavancin from the concentrate of g) in aqueous ethanol, and
i) drying the precipitated oritavancin, thereby preparing an oritavancin drug sub stance preparation,
wherein the oritavancin drug substance preparation has a maximum impurity level of not more than 3.0% by peak area of impurity 1 and impurity 7, defined by peak A of FIG. 1 and peak G of FIG. 2 , respectively; or
wherein the oritavancin drug substance preparation has about 85% purity or greater by peak area relative to impurities 1-16, defined by peak A of FIG. 1 and peaks B-P of FIG. 2 , respectively.