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Title: Scope and Limitations of Fmoc Chemistry SPPS-Based Approaches to the Total Synthesis of Insulin Lispro via Ester Insulin

Journal Article · · Chemistry - A European Journal
 [1];  [1];  [2];  [2];  [3];  [3];  [3];  [1]
  1. Univ. of Chicago, IL (United States)
  2. Case Western Reserve Univ., Cleveland, OH (United States)
  3. Bachem Holding AG, Bubendorf (Switzerland)
+ Show Author Affiliations

We have systematically explored three approaches based on 9-fluorenylmethoxycarbonyl (Fmoc) chemistry solid phase peptide synthesis (SPPS) for the total chemical synthesis of the key depsipeptide intermediate for the efficient total chemical synthesis of insulin. The approaches used were: stepwise Fmoc chemistry SPPS; the “hybrid method”, in which maximally protected peptide segments made by Fmoc chemistry SPPS are condensed in solution; and, native chemical ligation using peptide-thioester segments generated by Fmoc chemistry SPPS. A key building block in all three approaches was a Glu[O-β-(Thr)] ester-linked dipeptide equipped with a set of orthogonal protecting groups compatible with Fmoc chemistry SPPS. We report the most effective method for the preparation of the 51 residue ester-linked polypeptide chain of ester insulin was the use of unprotected peptide-thioester segments, prepared from peptide-hydrazides synthesized by Fmoc chemistry SPPS, and condensed by native chemical ligation. High-resolution X-ray crystallography confirmed the disulfide pairings and three-dimensional structure of synthetic insulin lispro prepared from ester insulin lispro by this route. Further optimization of these pilot studies could yield an efficient total chemical synthesis of insulin lispro (Humalog) based on peptide synthesis by Fmoc chemistry SPPS.

Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
RR-15301; AC02-06CH11357
OSTI ID:
1430310
Journal Information:
Chemistry - A European Journal, Vol. 23, Issue 7; ISSN 0947-6539
Publisher:
ChemPubSoc EuropeCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 10 works
Citation information provided by
Web of Science

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Cited By (4)

Insulin-From its Discovery to the Industrial Synthesis of Modern Insulin Analogues journal July 2017
Novel protein science enabled by total chemical synthesis journal December 2018
Characterization and optimization of two-chain folding pathways of insulin via native chain assembly journal May 2018
Sustainable, cost-efficient manufacturing of therapeutic peptides using chemo-enzymatic peptide synthesis (CEPS) journal January 2019

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Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Fmoc chemistry SPPS
hybrid synthesis
insulin lispro
native chemical ligation
total synthesis