Polyoxypeptins A and B Produced by Streptomyces: Apoptosis-Inducing Cyclic Depsipeptides Containing the Novel Amino Acid (2S,3R)-3-Hydroxy-3-methylproline

Abstract

Potent apoptosis-inducing peptides, polyoxypeptins A (1) and B (2) were isolated from the culture broth of Streptomyces sp. by solvent extraction and column chromatography. Structural elucidation of 1 by MS and NMR analyses revealed that it is a cyclic hexadepsipeptide having a novel amino acid and a previously unrecognized N-acyl side chain. The depsipeptide consisted of 3-hydroxyleucine, N-hydroxyvaline, N-hydroxyalanine, piperazic acid, 5-hydroxyhexahydropiperazine-3-carboxylic acid, and an unusual and hitherto unreported amino acid, 3-hydroxy-3-methylproline. Alanine and valine obtained from the hydrolyzate of 1 were both in the L-configuration as concluded from chiral TLC analysis. Then, the absolute structure of 1 was determined with the relative structure obtained from X-ray crystallographic analysis, and the new amino acid was isolated and confirmed to be (2S,3R)-3-hydroxy-3-methylproline (3). MS and NMR of 2 exhibited that it is a monodeoxy compound of 1. Stereochemistry of 2 was determined by degradation studies. Both 1 and 2 at a concentration of about 0.1 &mgr;g/mL induced early cell death, nuclear fragmentation, and internucleosomal DNA scission, all of which are characteristic of apoptosis, in human pancreatic carcinoma AsPC-1 cells.