title: Directed Evolution of Scanning Unnatural-Protease-Resistant (SUPR) Peptides for in Vivo Applications.
abstract: Peptides typically have poor biostabilities, and natural sequences cannot easily be converted into drug-like molecules without extensive medicinal chemistry. We have adapted mRNA display to drive the evolution of highly stable cyclic peptides while preserving target affinity. To do this, we incorporated an unnatural amino acid in an mRNA display library that was subjected to proteolysis prior to selection for function. The resulting "SUPR (scanning unnatural protease resistant) peptide" showed ≈500-fold improvement in serum stability (t1/2 =160 h) and up to 3700-fold improvement in protease resistance versus the parent sequence. We extended this approach by carrying out SUPR peptide selections against Her2-positive cells in culture. The resulting SUPR4 peptide showed low-nanomolar affinity toward Her2, excellent specificity, and selective tumor uptake in vivo. These results argue that this is a general method to design potent and stable peptides for in vivo imaging and therapy.
authors.affiliation: Department of Chemistry, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.|EvoRx Technologies, 129 N. Hill Avenue, Suite 103, Pasadena, CA, 91106, USA.
authors.forename: Stephen V
authors.initials: SV
authors.lastname: Fiacco
authors.affiliation: Department of Cancer Systems Imaging, University of Texas, MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA.
authors.forename: Lindsay E
authors.initials: LE
authors.lastname: Kelderhouse
authors.affiliation: EvoRx Technologies, 129 N. Hill Avenue, Suite 103, Pasadena, CA, 91106, USA.
authors.forename: Amanda
authors.initials: A
authors.lastname: Hardy
authors.affiliation: Department of Chemistry, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.
authors.forename: Yonatan
authors.initials: Y
authors.lastname: Peleg
authors.affiliation: Mork Family Department of Chemical Engineering and Materials Science, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.
authors.forename: Biliang
authors.initials: B
authors.lastname: Hu
authors.affiliation: Department of Cancer Systems Imaging, University of Texas, MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA.
authors.forename: Argentina
authors.initials: A
authors.lastname: Ornelas
authors.affiliation: Integrative Medicine Program, University of Texas, MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA.
authors.forename: Peiying
authors.initials: P
authors.lastname: Yang
authors.affiliation: Department of Cancer Systems Imaging, University of Texas, MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA.
authors.forename: Seth T
authors.initials: ST
authors.lastname: Gammon
authors.affiliation: Department of Chemistry, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.
authors.forename: Shannon M
authors.initials: SM
authors.lastname: Howell
authors.affiliation: Mork Family Department of Chemical Engineering and Materials Science, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.|University of Southern California, Norris Comprehensive Cancer Center, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.
authors.forename: Pin
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA.
authors.forename: Terry T
authors.initials: TT
authors.lastname: Takahashi
authors.affiliation: Department of Cancer Systems Imaging, University of Texas, MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA. smillward@mdanderson.org.|Department of Bioengineering, Rice University, 6100 Main Street, Houston, TX, 77005, USA. smillward@mdanderson.org.
authors.forename: Steven W
authors.initials: SW
authors.lastname: Millward
authors.affiliation: Department of Chemistry, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA. richrob@usc.edu.|Mork Family Department of Chemical Engineering and Materials Science, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA. richrob@usc.edu.|Department of Molecular and Computational Biology, University of Southern California, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA. richrob@usc.edu.|University of Southern California, Norris Comprehensive Cancer Center, 3710 McClintock Avenue, Los Angeles, CA, 90089-1211, USA. richrob@usc.edu.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Roberts
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D019020:Directed Molecular Evolution; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D010447:Peptide Hydrolases / Q000378:metabolism*; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D055550:Protein Stability; D012333:RNA, Messenger / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D000596:Amino Acids; D019151:Peptide Library; D010456:Peptides, Cyclic; D012333:RNA, Messenger; D010447:Peptide Hydrolases
keywords: Her2 receptor; directed evolution; mRNA display; peptides; unnatural amino acids