Methods of Identifying a Compound that Increases or Decreases the Interaction of Hn-33 with Synaptotagmin II

Abstract

This application is based, inter alia, on the discovery of a binding interaction between the Hn-33 hemagglutinin polypeptide of the type A Clostridium botulinum neurotoxin complex and synaptosomal proteins, including synaptotagmin II (Syt II). Methods of screening for compounds that modulate, e.g., increase or decrease, this interaction are provided. Also provided are compositions and methods for targeting compounds to neuronal and cancer cells by coupling the compounds to Hn-33 or biologically active Hn-33 variants.

1 - 7 . (canceled) 
     
     
         8 . A composition comprising an Hn-33 polypeptide or a synaptotagmin-binding Hn-33 polypeptide variant coupled to a non-botulinum neurotoxin compound. 
     
     
         9 . The composition of  claim 8 , wherein the Hn-33 polypeptide or Hn-33 polypeptide variant binds to a cell that expresses a synaptotagmin protein. 
     
     
         10 . The composition of  claim 8 , wherein the non-botulinum neurotoxin compound is a polypeptide and the Hn-33 polypeptide or Hn-33 polypeptide variant provides protease resistance to the polypeptide. 
     
     
         11 . The composition of  claim 9 , wherein the synaptotagmin protein is synaptotagmin II. 
     
     
         12 . The composition of  claim 9 , wherein the cell is a neuronal cell or a cancer cell. 
     
     
         13 . The composition of  claim 12 , wherein the cancer cell is a neuronal cancer cell. 
     
     
         14 . The composition of  claim 12 , wherein the non-botulinum neurotoxin compound is a polypeptide. 
     
     
         15 . The compound of  claim 14 , wherein the non-botulinum neurotoxin polypeptide is an antibody or a biologically active fragment thereof. 
     
     
         16 . The composition of  claim 8 , wherein the composition is a therapeutic composition. 
     
     
         17 . The composition of  claim 9 , wherein the compound is a small molecule. 
     
     
         18 . The composition of  claim 17 , wherein the small molecule is a neuroactive drug. 
     
     
         19 . The composition of  claim 19 , wherein the compound is a chemotherapeutic agent. 
     
     
         20 - 37 . (canceled) 
     
     
         38 . Use of a composition of  claim 9  for the manufacture of a medicament for the treatment of cancer. 
     
     
         39 . Use of a composition of  claim 9  for the manufacture of a medicament for targeting the non-botulinum neurotoxin compound to a neuronal cell in a subject. 
     
     
         40 . Use of a composition comprising an inhibitor of an interaction between Hn-33 and a synaptosomal protein for the treatment of a subject afflicted with botulinum poisoning. 
     
     
         41 . The use of  claim 40 , wherein the synaptosomal protein is synaptotagmin II. 
     
     
         42 . The use of  claim 40 , wherein the inhibitor is selected from the group consisting of an antibody that binds to Hn-33, an antibody that binds to the synaptosomal protein, an Hn-33 antibody anti-idiotype antibody, a biologically active Hn-33 variant, and a biologically active synaptosomal protein variant.