Methods and compositions for controlled polypeptide synthesis

• US 20020032309A1
• Filed: 06/08/2001
• Published: 03/14/2002
• Est. Priority Date: 03/19/1998
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Wyrsta, Michael D., Deming, Timothy J., Curtin, Scott A., Seidel, Scott W., Hwang, Jungyeon, Yu, Miaoer, Nowak, Andrew
• Granted Patent: US 6,686,446 B2
• Time in Patent Office: Days
• US Class Current: 530/350
• CPC Class Codes: C07K 1/084 containing nitrogen
• CPC Class Codes: C07K 1/088 containing other elements, e.g. B, Si, As
• CPC Class Codes: C07K 1/10 using coupling agents C07K1/006 takes precedence
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C08G 69/10 Alpha-amino-carboxylic acids polysuccinimides C08G73/1092

Abstract

Methods and compositions for the generation of polypeptides having varied material properties are disclosed herein. Methods include means for initiating the polymerization of aminoacid-N-carboxyanhydride (NCA) monomer by combining the monomer with an amido-containing metallacycle, for making self assembling amphiphilic block copolypeptides and related protocols for adding oligo(ethyleneglycol) functionalized aminoacid-N-carboxyanhydrides (NCAs) to polyaminoacid chains. Additional methods include means of adding an end group to the carboxy terminus of a polyaminoacid chain by reacting an alloc-protected amino acid amide with a transition metal-donor ligand complex to forming an amido-amidate metallacycle for use in further polymerization reactions. Novel compositions for use in peptide synthesis and design including five and six membered amido-containing metallacycles and block copolypeptides are also disclosed.

Claims

1. A method of initiating an a-amino acid-N-carboxyanhydride monomer polymerization comprising combining an NCA monomer with an initiator molecule comprising an amido-containing metallacycle, which contains a nucleophilic alkyl amido group stabilized by a rigid chelate and a non-nucleophilic proton-accepting group, wherein the proton-accepting group is selected from the group of amido sulfonamidate, an amido-amidate having an extracyclic nitrogen, an amido-ureate, and amido-carbamate, or an amido-aldimate.

2. The method of claim 1 wherein the initiator molecule contains a low valent transition metal.

3. The method of claim 2 wherein the the low valent transition metal is ruthenium.

4. The method of claim 1 wherein the initiator molecule is of general formula I, II, III or IV:

5. The method of claim 4 wherein R2 and R6 are hydrogen and R3 and R5 are phenyl.

6. The method of claim 4 wherein the Lewis base ligand of the initiator molecule is p-cymene.

7. The method of claim 1 wherein the initiator molecule is of the following general formula:

8. A polyaminoacid chain comprising at least ten consecutive oligo(ethyleneglycol)-conjugated amino acid resdues.

9. An amphiphilic block copolypeptide comprising a soluble block polypeptide and and an insoluble block polypeptide, said soluble block having at least about 30% mole percent identical amino acid residues having charged or oligo(ehtyleneglycol)-conjugated side chains and said insoluble block comprising at about 60 to 100 mole percent nonionic amino acid residues.

10. The amphiphilic block copolypeptide of claim 9 wherein the insoluble block comprises about 3 to about 60 mole percent of the total copolypeptide.

11. The amphiphilic block copolypeptide of claim 9 wherein the nonionic amino acid residues are selected from the group consisting of phenylalanine, leucine, valine, isoleucine, alanine and methionine.

12. The amphiphilic block copolypeptide of claim 9 wherein the amino acid residues having charged side chains are selected from the group consisting of glutamic acid, aspartic acid, arginine, histidine, lysine, and ornithine.

13. The amphiphilic block copolypeptide of claim 9 wherein the amino acid residues having oligo(ehtyleneglycol)-conjugated side chains are selected from the group consisting of EG-cysteine, EG-lysine, EG-serine, and EG-tyrosine.

14. A chain-end functionalized block polypeptide having ten or more consecutive identical amino acid residues and an endgroup selected from the group consisting of an oligosaccharide, oligonucleotide, fluoresent molecule, polymer chain, small molecule therapeutic, or reactive chemical linker to attach the block copolypeptide to another molecule.

15. A chain-end functionalized block copolypeptide having an end group selected from the group consisting of a napthyl group, an alkyl group, an allyl group, and cysteinamide.