Compositions and methods for altering cocaine esterase activity
The present invention relates to compositions and methods for treating and preventing cocaine addiction. In particular, the present invention provides mutated cocaine esterase proteins for use in treating and preventing cocaine addiction.
1. A composition comprising a mutant cocaine esterase (CocE) polypeptide, wherein said cocaine esterase polypeptide has at least one mutation selected from the group consisting of G4C and S10C relative to the CocE polypeptide of SEQ ID NO:1 at the dimer interface, and wherein said mutation stabilizes the dimer interface between monomers of said CocE.
2. The composition of claim 1 , wherein said mutant CocE polypeptide has an increased half life relative to wild type CocE.
3. The composition of claim 2 , wherein said half life is at least 1 hour.
4. The composition of claim 2 , wherein said half life is at least 1 day.
5. The composition of claim 2 , wherein said half life is at least 3 days.
6. The composition of claim 1 , wherein said mutant CocE polypeptide has G4C and S10C mutations relative to the CocE polypeptide of SEQ ID NO:1.
7. The composition of claim 1 , wherein said mutant CocE polypeptide further comprises as least one additional mutation that stabilizes inter-domain or intra-domain contacts in domain II of said polypeptide.
8. The composition of claim 7 , wherein said additional mutation is selected from the group consisting of T172R, G173Q and L169K relative to the CocE polypeptide of SEQ ID NO:1.
9. The composition of claim 8 , wherein said additional mutation is selected from the group consisting of T172R+G172Q and L169K+G173Q relative to the CocE polypeptide of SEQ ID NO:1.
10. The composition of claim 8 , wherein said mutant CocE polypeptide has G4C, S10C, L169K and G173Q mutations relative to the CocE polypeptide of SEQ ID NO:1.
11. The composition of claim 8 , wherein said mutant CocE polypeptide has G4C, S10C, T172R and G173Q mutations relative to the CocE polypeptide of SEQ ID NO:1.
12. The composition of claim 1 , wherein said mutant CocE polypeptide is PEGylated.
13. The composition of claim 12 , wherein said CocE mutant polypeptide has A92C and S99C mutations relative to the CocE polypeptide further of SEQ ID NO:1.
14. The composition of claim 13 , wherein said CocE mutant polypeptide further has C429S and C477S mutations relative to the CocE polypeptide of SEQ ID NO:1.
15. A method of reducing at least one biological activity of cocaine, comprising: administering a pharmaceutical composition comprising a mutant cocaine esterase (CocE) polypeptide, wherein said cocaine esterase polypeptide has at least one mutation at the dimer interface selected from the group consisting of G4C and S10C relative to the CocE polypeptide of SEQ ID NO:1, and wherein said mutation stabilizes the dimer interface between monomers of said CocE to a subject that has previously ingested cocaine or is likely to ingest cocaine.
16. The method of claim 15 , wherein said subject is addicted to cocaine.
17. The method of claim 15 , wherein said subject has ingested an overdose of cocaine.
18. The method of claim 15 , wherein said pharmaceutical composition is administered once.
19. The method of claim 15 , wherein said pharmaceutical composition is administered more than once.
20. A composition comprising a nucleic acid encoding a mutant cocaine esterase (CocE) polypeptide, wherein said cocaine esterase polypeptide has at least one mutation at the dimer interface selected from the group consisting of G4C and S10C relative to the CocE polypeptide of SEQ ID NO:1, and wherein said mutation stabilizes the dimer interface between monomers of said CocE.
21. The composition of claim 20 , wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 12 and 14.