Process for preparing dipeptidyl peptidase IV inhibitors and intermediates therefor
AssigneeBristol-Myers Squibb Company
InventorsMichael Politino, Matthew M. Cadin, Paul M. Skonezny, Jason G. Chen
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Abstract
A process for production of cyclopropyl-fused pyrrolidine-based inhibitors of dipeptidyl peptidase IV is provided which employs a BOC-protected amine of the structure prepared by subjecting an acid of the structure to reduce amination by treating the acid with ammonium formate, nicotinamide adenine dinucleotide, dithiothreitol and partially purified phenylalanine dehydrogenase/formate dehydrogenase enzyme concentrate (PDH/FDH) and without isolating treating the resulting amine of the structure 2 with di-tert-butyl dicarbonate to form the BOC-protected amine.
Claims9
A process of preparing a partially purified enzyme concentrate of phenylalanine dehydrogenase (PDH) and/or formate dehydrogenase (FDH), the process comprising:
- a. preparing a fermentation broth containing a microorganism capable of producing PDH and/or FDH;
- b. microfluidizing the fermentation broth while maintaining the temperature of said fermentation broth between about 4° C. and 30° C. to form a microfluidized broth containing the PDH and/or FDH;
- c. clarifying the microfluidized broth by treating the broth with a flocculating agent to coagulate cell debris and to remove DNA and unwanted proteins, thereby forming a clarified broth;
- d. filtering the clarified broth to give a filtrate;
- and
- e. concentrating the filtrate to give said partially purified enzyme concentrate, the concentrate containing:
- i. said PDH having a PDH activity of about 400 IU/ml to about 1000 IU/ml;
- and/or
- ii. said FDH having a FDH activity of about 20 IU/ml to about 200 IU/ml,
- wherein the enzyme concentrate is capable of reductively aminating a keto-containing compound into a chiral amine-containing compound;
- wherein said chiral amine-containing compound, without isolation from the enzyme concentrate, is capable of being BOC-protected;
- and wherein, optionally, the clarifying comprises contacting the microfluidized broth with diatomaceous earth and the filtering comprises filtering the diatomaceous earth with a filter press.
Dependencies are inferred from the claim text.