Cocaine hydrolase-FC fusion proteins for cocaine and methods for utilizing the same
The presently-disclosed subject matter includes isolated polypeptides that comprise a butyrylcholinestrase (BChE) polypeptide and a second polypeptide. The BChE polypeptide as well as the second polypeptide can be variants and/or fragments thereof. The presently-disclosed subject matter also includes a pharmaceutical composition that comprises the present isolated polypeptide and a suitable pharmaceutical carrier. Further still, methods are provided for treating cocaine-induced conditions, and comprise administering the isolated polypeptide and/or pharmaceutical compositions thereof to an individual.
1. An isolated polypeptide having the ability to cleave cocaine, comprising
a butyrylcholinesterase (BChE) polypeptide having the ability to cleave cocaine, consisting of a variant of SEQ ID NO: 26 or a fragment thereof; and
a second polypeptide, consisting of the sequence of SEQ ID NO: 18;
wherein the variant of SEQ ID NO: 26 or the fragment thereof includes between 5 and 8 amino acid substitutions;
wherein 5 of the amino acid substitutions consist of A199S, F227A, S287G, A328W, and Y332G.
2. The isolated polypeptide of claim 1 , wherein the isolated polypeptide having the ability to cleave cocaine comprises an amino acid sequence selected from the group of sequences consisting of: SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 178, SEQ ID NO: 180, and SEQ ID NO: 182.
3. The isolated polypeptide of claim 1 , wherein the BChE polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 48, 50, 58, and 62.
4. A pharmaceutical composition comprising:
an isolated polypeptide of claim 1 ; and
a suitable pharmaceutical carrier.
5. The pharmaceutical composition of claim 4 , wherein, from the N-terminus to the C-terminus, the isolated polypeptide comprises the BChE polypeptide and the second polypeptide.
6. The isolated polypeptide of claim 2 , wherein the amino acid sequence is selected from the group of sequences consisting of: SEQ ID NO: 169, SEQ ID NO:
180, and SEQ ID NO: 182.
7. The isolated polypeptide of claim 1 , wherein, in addition to A199S, F227A, S287G, A328W, and Y332G, the variant includes from 1 to 3 amino acid substitutions selected from the group consisting of P285A, P285E, P285G, P285I, P285K, P285N, P285Q, P285S, L286M, and E441D.