Rapamycin and process of preparation

• US 3,929,992 A
• Filed: 04/12/1974
• Issued: 12/30/1975
• Est. Priority Date: 09/29/1972
• Status: Expired due to Term

• Current Assignee: Claude Vezina, Surendra N. Sehgal, Teodora M. Blazekovic
• Original Assignee: Claude Vezina, Surendra N. Sehgal, Teodora M. Blazekovic
• Inventors: Sehgal, Surendra N., Blazekovic, Teodora M., Vezina, Claude
• Primary Examiner(s): Goldberg, Jerome D.
• Application Number:
  US05/460,665
• Time in Patent Office: 627 Days
• Field of Search: 424/122 195/80
• US Class Current: 424/122
• CPC Class Codes: C12P 17/188 Heterocyclic compound containing in the condensed system at least one hetero ring having nitrogen atoms and oxygen atoms as the only ring heteroatoms ergot-alcaloids C12P17/183
• CPC Class Codes: Y10S 435/898 Streptomyces hygroscopicus

Abstract

Antibiotic rapamycin is producible by culturing Streptomyces hygroscopicus NRRL 5491 in an aqueous nutrient medium. Rapamycin has antifungal properties. Methods for its preparation and use are disclosed.

Claims

1. RAPAMYCIN, AN ANTIBIOTIC WHICH A. IS A LOLOURLESS, CRYSTALLINE COMPOUND WITH A MELTING POINT OF 183* TO 185*C, AFTER RECRYSTALLIZATION FROM ETHER;

B. IS SOLUBLE IN ETHER, CHLOROFORM, ACETONE, METHANOL AND DIMETHYLFORMAMIDE, VERY SPARINGLY SOLUBLE IN HEXANE AND PETROLEUM ETHER AND SUBSTANTIALLY INSOLUBLE IN WATER;

C. SHOWS A UNIFORM SPOT ON THIN LAYER PLATES OF SILICA GEL;

D. HAS A CHARACTERISTIC ELEMENTAL ANALYSIS OF ABOUT C. 66.84%, H, 8.84%;

N, 1.37%, E. EXHIBITS THE FOLLOWING CHARACTERISTIC ABSORPTION MAXIMA IN ITS ULTRAVIOLET ABSORPTION SPECTRUM (95% ETHANOL) 267 NM (E1CM1%

     417), 277 NM (E1CM1%

     541) AND 288 NM (E1CM1%

     416);

F. HAS A CHARACTERISTIC INFRARED ABSORPTION SPECTRUM SHOWN IN ACCOMPANYING FIG. 1;

G. HAS A CHARACTERISTIC NUCLEAR MAGNETIC RESONANCE SPECTRUM AS SHOWN IN ACCOMPANYING FIG. 2;

H. HAS A MINIMUM INHIBITORY CONCENTRATION OF 0.002 TO 0.1 UG/ML AGAINST CANDIDA ALBICANS;

AND I. EXHIBITS A LD50 (I.P., MICE) OF 597.3 $ 28.1 MG/KG AND A LD50 (P.O., MICE) OF >

2,500 MG/KG.

2. A process for the production of rapamycin which comprises cultivating Streptomyces hygroscopicus NRRL 5491 in an aqueous nutrient medium containing a source of assimilable carbon and nitrogen and mineral salts under aerobic conditions until substantial antifungal activity is present in the fermentation mixture by the production of rapamycin, and isolating rapamycin from said fermentation mixture.

3. A process as claimed in claim 2 in which the cultivation is carried out at a temperature ranging from 20* to 35*C and at an initial pH of between 6.5 and 7.5.

4. A process according to claim 2 in which the isolation comprises filtering the fermentation mixture, extracting the filter cake with methanol or ethanol to produce an extract, and separating the rapamycin from the extract.