Chromatographic purification process

• US 4,874,843 A
• Filed: 12/03/1987
• Issued: 10/17/1989
• Est. Priority Date: 12/03/1987
• Status: Expired due to Term

• Current Assignee: Eli Lilly and Company
• Original Assignee: Eli Lilly and Company
• Inventors: Baker, Patrick J.
• Primary Examiner(s): LEE, LESTER
• Application Number:
  US07/128,351
• Time in Patent Office: 684 Days
• Field of Search: 530/317, 530/322, 530/344
• US Class Current: 530/317
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C07K 7/56 the cyclisation not occurring through 2,4-diamino-butanoic acid

Abstract

This invention relates to a new chromatographic process for purifying fermentation products, particularly the antibiotic LY146032, from fermentation broths by use of a reverse phase non-functional resin.

In the process of this invention the resin is loaded with the compound in the aqueous phase, the water then is removed from the resin and the resin converted to the organic phase for the resolution step. This process is applicable to purifying compounds which are adsorbed on reverse phase non-functional resins.

Claims

1. A process for purifying fermentation products which comprises:

a. contacting an aqueous solution of a fermentation product with a non-functionalized macroporous copolymer of styrene and divinylbenzene in aqueous phase;

b. physically removing the water from the charged resin;

c. rewetting the charged resin with a polar organic solvent;

d. washing the resin with a polar organic solvent that is the same as or different than the polar organic solvent used in step (C) to remove non-uv impurities;

e. eluting the fermentation product from the non-functional resin by increasing the polarity of the solvent; and

f. recovering the fermentation product.

2. The process of claim 1 wherein the fermentation product is a lipopeptide.

3. The process of claim 2 wherein the fermentation product is LY146032.

4. The process of claim 2 wherein the fermentation product is Echinocandin B.

5. The process of claim 1 wherein the fermentation product is a glycopeptide.

6. The process of claim 4 wherein the fermentation product is vancomycin.

7. The process of claim 1 wherein the resin is Diaion HP-20.

8. The process of claim 1 wherein the resin is amberlite XAD-4.

9. The process of claim 1 wherein the polar organic solvent is acetonitrile.

10. The process of claim 1 wherein the fermentation product is eluted from the non functionalized macroporous copolymer of styrene and divinylbenzene with acetonitrile containing from about 5% to about 60% water.

11. The process of claim 9 wherein the fermentation product is eluted from the non-functionalized macroporous copolymer of styrene and divinylbenzene with acetonitrile containing from about 10% to about 20% water.