Methods for preparing high throughput peptidomimetics, orally bioavailable drugs and compositions containing same
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. A non-naturally occurring peptide comprising:
MA X Y X HYHK, (SEQ. ID NO: 81) wherein the X at positions 3 and 5 is N-methyl norvaline; or
MVC XX LYDDK, (SEQ ID NO: 82) wherein the X at positions 4 and 5 is N-methyl norvaline.
2. A non-naturally occurring peptide of claim 1 , wherein the peptide is biotinylated at the N-terminal or C-terminal end.
3. A peptide conjugate comprising a non-naturally occurring peptide of claim 1 , wherein the peptide is conjugated to polyethylene glycol or a lipid molecule.
4. A composition comprising a non-naturally occurring peptide of claim 1 and a carrier.
5. The composition of claim 4 , wherein the carrier is a pharmaceutically acceptable carrier.
6. A method for inhibiting the growth of a breast cancer cell, comprising contacting the cell with an effective amount of a non-naturally occurring peptide of claim 1 , wherein the breast cancer cell is HER2+.
7. A method for treating breast cancer in subject in need thereof, comprising administering to the subject an effective amount of a non-naturally occurring peptide of claim 1 , wherein the breast cancer cell is HER2+.
8. A method for detecting HER2+ a breast cancer cell in a subject comprising administering to the subject a non-naturally occurring peptide of claim 1 and screening for the presence of any of the peptide bound to a HER2+ breast cancer cell in the subject.
9. The method of claim 8 , wherein the non-naturally occurring peptide is detectably labeled.
10. The method of claim 9 , wherein the label is a fluorescent dye or a PET label.
11. A vector comprising an isolated nucleic acid encoding the peptide of claim 1 , wherein N-methyl norvaline is encoded by a stop codon.
12. A method for producing a non-naturally occurring peptide of claim 1 comprising translating an isolated nucleic acid encoding said peptide in rabbit reticulocyte lysate,
wherein said lysate is supplemented with a suppressor tRNA charged with N-methyl norvaline, and
wherein N-methyl norvaline is encoded by a stop codon complimentary to said suppressor tRNA.
13. The method of claim 12 , further comprising isolating the non-naturally occurring peptide from the lysate.
14. A method for determining if a candidate agent is a potential therapeutic to inhibit the growth of a HER2+ breast cancer cell, comprising: contacting the candidate agent with a breast cancer cell and assaying for growth inhibitory activity, and comparing the inhibitory activity of the candidate agent with the inhibitory activity of: the non-naturally occurring peptide of claim 1 .
15. A kit for one or more of: inhibiting the growth of a breast cancer cell or for treating breast cancer, or for determining if a candidate agent is a potential therapeutic to inhibit the growth of a cancer cell, comprising the non-naturally occurring peptide of claim 1 and instructions for use.