title: A general method for making peptide therapeutics resistant to serine protease degradation: application to dipeptidyl peptidase IV substrates.
abstract: Bioactive peptides have evolved to optimally fulfill specific biological functions, a fact which has long attracted attention for their use as therapeutic agents. While there have been some recent commercial successes fostered in part by advances in large-scale peptide synthesis, development of peptides as therapeutic agents has been significantly impeded by their inherent susceptibility to protease degradation in the bloodstream. Here we report that incorporation of specially designed amino acid analogues at the P1' position, directly C-terminal of the enzyme cleavage site, renders peptides, including glucagon-like peptide-1 (7-36) amide (GLP-1) and six other examples, highly resistant to serine protease degradation without significant alteration of their biological activity. We demonstrate the applicability of the method to a variety of proteases, including dipeptidyl peptidase IV (DPP IV), dipeptidyl peptidase 8 (DPP8), fibroblast activation protein α (FAPα), α-lytic protease (αLP), trypsin, and chymotrypsin. In summary, the "P1' modification" represents a simple, general, and highly adaptable method of generating enzymatically stable peptide-based therapeutics.
authors.affiliation: Department of Biochemistry, Tufts University Sackler School of Graduate Biomedical Sciences , 136 Harrison Avenue, Boston, Massachusetts 02111, United States.
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Heard
authors.forename: Wengen
authors.initials: W
authors.lastname: Wu
authors.forename: Youhua
authors.initials: Y
authors.lastname: Li
authors.forename: Peng
authors.initials: P
authors.lastname: Zhao
authors.forename: Iwona
authors.initials: I
authors.lastname: Woznica
authors.forename: Jack H
authors.initials: JH
authors.lastname: Lai
authors.forename: Martin
authors.initials: M
authors.lastname: Beinborn
authors.forename: David G
authors.initials: DG
authors.lastname: Sanford
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Dimare
authors.forename: Amrita K
authors.initials: AK
authors.lastname: Chiluwal
authors.forename: Diane E
authors.initials: DE
authors.lastname: Peters
authors.forename: Danielle
authors.initials: D
authors.lastname: Whicher
authors.forename: James L
authors.initials: JL
authors.lastname: Sudmeier
authors.forename: William W
authors.initials: WW
authors.lastname: Bachovchin
mesh_terms: D000818:Animals; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008820:Mice, Obese; D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D015842:Serine Proteinase Inhibitors; D018819:Dipeptidyl Peptidase 4