title: Production of novel rapamycin analogs by precursor-directed biosynthesis.
abstract: The natural product rapamycin, produced during fermentation by Streptomyces hygroscopicus, is known for its potent antifungal, immunosuppressive, and anticancer activities. During rapamycin biosynthesis, the amino acid l-pipecolate is incorporated into the rapamycin molecule. We investigated the use of precursor-directed biosynthesis to create new rapamycin analogs by substitution of unusual l-pipecolate analogs in place of the normal amino acid. Our results suggest that the l-pipecolate analog (+/-)-nipecotic acid inhibits the biosynthesis of l-pipecolate, thereby limiting the availability of this molecule for rapamycin biosynthesis. We used (+/-)-nipecotic acid in our precursor-directed biosynthesis studies to reduce l-pipecolate availability and thereby enhance the incorporation of other pipecolate analogs into the rapamycin molecule. We describe here the use of this method for production of two new sulfur-containing rapamycin analogs, 20-thiarapamycin and 15-deoxo-19-sulfoxylrapamycin, and report measurement of their binding to FKBP12.
authors.affiliation: Natural Products, Wyeth Research, 401 North Middletown Road, Building 205, Room 465, Pearl River, NY 10965, USA. ritaccf@wyeth.com
authors.forename: Frank V
authors.initials: FV
authors.lastname: Ritacco
authors.forename: Edmund I
authors.initials: EI
authors.lastname: Graziani
authors.forename: Mia Y
authors.initials: MY
authors.lastname: Summers
authors.forename: T Mark
authors.initials: TM
authors.lastname: Zabriskie
authors.forename: Ker
authors.initials: K
authors.lastname: Yu
authors.forename: Valerie S
authors.initials: VS
authors.lastname: Bernan
authors.forename: Guy T
authors.initials: GT
authors.lastname: Carter
authors.forename: Michael
authors.initials: M
authors.lastname: Greenstein
mesh_terms: D001709:Biotechnology / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D009557:Nipecotic Acids / Q000378:metabolism; D010875:Pipecolic Acids / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000378:metabolism*; D013302:Streptomyces / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D022061:Tacrolimus Binding Protein 1A / Q000378:metabolism
chemical_list: D009557:Nipecotic Acids; D010875:Pipecolic Acids; D011498:Protein Precursors; C030278:nipecotic acid; D022061:Tacrolimus Binding Protein 1A; C031345:pipecolic acid; D020123:Sirolimus