Methods for preparing high throughput peptidomimetics, orally bioavailable drugs and compositions containing same

Abstract

Provided herein are methods to generate and screen peptides that exhibit drug like stabilities in vitro and in vivo. By selecting for enzyme resistance, Applicants are able to derive peptides that are not only stable to a broad spectrum of proteases, but also stable to other drug processing enzymes such as cytochrome P450s. This approach provides a general method to the rapid development of highly stable peptides for therapeutic development and diagnosis. The peptides are further modified for oral bioavailability. The methods can be applied to similar peptides for the making of therapeutic compositions

1 - 7 . (canceled) 
     
     
         8 . A method for generating one or stable, bioavailable peptide(s), comprising:
 a. mutating a library of nucleic acids encoding peptides selected for a pre-determined specificity to incorporate one or more amino acids that impart stability;   b. incorporating one or more stop codons;   c. supplementing the library with one or more tRNA charged with an amino acid that imparts stability to suppress the stop codon;   d. translating one or more of the individual sequences of the nucleic library to one or more peptides;   e. cyclizing the one or more peptides; and   f. selecting individual peptides for protease resistance   wherein the amino acid that imparts stability is selected from the group of: an 2-aminoisobutyric acid (AIB), a D-amino acid, a beta (B) amino acid, and an N-methyl amino acid.   
     
     
         9 . The method of  claim 8 , wherein the nucleic acid library is a DNA library or RNA library. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 8 , further comprising:
 g. conjugating the peptide to a biotin molecule or biotin analog through a linkage.   
     
     
         12 . The method of  claim 11 , wherein the biotin molecule or biotin analog is one or more of: is a reducible biotin molecule; a biotin on the side chain of lysine of the peptide; a biotin linked to the peptide through an amide linkage or an ester linkage, a biotin linked to the peptide through a thioester linkage or an ester linkage. 
     
     
         13 . (canceled) 
     
     
         14 . The method of  claim 8 , wherein the peptide selected for a predetermined specificity is pre-selected by a method selected from the group of: specificity against a cell surface protein, and/or specificity against mammalian cell culture or tissue; and/or specificity against a purified protein target. 
     
     
         15 - 16 . (canceled) 
     
     
         17 . The method of  claim 14 , wherein the purified protein target is a target of a scanning unnatural protease resistant peptide (“SUPR”). 
     
     
         18 . (canceled) 
     
     
         19 . The method of  claim 8 , wherein the peptide selected for a pre-determined specificity is pre-selected for an in vivo therapeutic utility. 
     
     
         20 - 21 . (canceled) 
     
     
         22 . An isolated peptide generated according to the method of  claim 8 . 
     
     
         23 . (canceled) 
     
     
         24 . The composition of  claim 64 , wherein the carrier is a pharmaceutically acceptable carrier. 
     
     
         25 - 36 . (canceled) 
     
     
         37 . A vector comprising an isolated nucleic encoding the isolated peptide of  claim 22 . 
     
     
         38 . An isolated host cell comprising an isolated nucleic acid encoding the isolated peptide of  claim 22 . 
     
     
         39 . A method for producing a non-naturally occurring peptide comprising growing one or more an isolated host cells of  claim 38  under conditions that allow expression of the isolated nucleic acid. 
     
     
         40 . The method of  claim 39 , further comprising isolating the non-naturally occurring peptide from the isolated host cells or cell supernatant. 
     
     
         41 - 61 . (canceled) 
     
     
         62 . The method of  claim 8 , wherein said N-methyl amino acid is N-methyl alanine. 
     
     
         63 . The method of  claim 8 , wherein said in vivo therapeutic utility is selected from the group of: the ability to inhibit the growth or kill a cancer cell, the ability to inhibit or kill a precancerous cell, and to bind to a cell expressing a desired target receptor. 
     
     
         64 . A composition comprising the isolated peptide of  claim 22  and a carrier. 
     
     
         65 . An isolated peptide library comprising a plurality of peptides generated according to the method of  claim 8 . 
     
     
         66 . An isolated nucleic acid library comprising isolated nucleic acids encoding each of a plurality of peptides generated according to the method of  claim 8 . 
     
     
         67 . The isolated nucleic acid library of  claim 66 , wherein the nucleic acid library is a DNA library or RNA library.