US2014065126A1PendingUtilityA1

BChE ALBUMIN FUSIONS FOR THE TREATMENT OF COCAINE ABUSE

Assignee: SKLAIR-TAVRON LIORAPriority: Dec 8, 2009Filed: Sep 4, 2013Published: Mar 6, 2014
Est. expiryDec 8, 2029(~3.4 yrs left)· nominal 20-yr term from priority
C07K 14/765C07K 14/001A61P 25/30A61P 25/36C07K 2319/31C12Y 301/01084C12N 9/18A61K 38/00
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Claims

Abstract

A method of attenuating a biological effect of cocaine exposure in a primate. Such method includes administering to the primate an amount of a BChE-albumin fusion protein comprising the amino acid substitutions A227S, S315G, A356W, and Y360G, wherein the amount of the fusion protein is effective to cause attenuation of the biological effect of cocaine exposure in the primate.

Claims

exact text as granted — not AI-modified
1 . A method of attenuating a biological effect of a cocaine exposure in a primate comprising administering to the primate an amount of a fusion protein comprising:
 (a) a mutant butyrylcholinesterase (BChE) polypeptide comprising the sequence   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   EDDIIIATKNGKVRGMNLTVFGGTVTAFLGIPYAQPPLGRLRFKKPQSLT 
                 
                     
                 
                   KWSDIWNATKYANSCCQNIDQSFPGFHGSEMWNPNTDLSEDCLYLNVWIP 
                 
                     
                 
                   APKPKNATVLIWIYGGGFQTGTSSLHVYDGKFLARVERVIVVSMNYRVGA 
                 
                     
                 
                   LGFLALPGNPEAPGNMGLFDQQLALQWVQKNIAAFGGNPKSVTLFGESSG 
                 
                     
                 
                   AASVSLHLLSPGSHSLFTRAILQSGSFNAPWAVTSLYEARNRTLNLAKLT 
                 
                     
                 
                   GCSRENETEIIKCLRNKDPQEILLNEAFVVPYGTPLGVNFGPTVDGDFLT 
                 
                     
                 
                   DMPDILLELGQFKKTQILVGVNKDEGTWFLVGGAPGFSKDNNSIITRKEF 
                 
                     
                 
                   QEGLKIFFPGVSEFGKESILFHYTDWVDDQRPENYREALGDVVGDYNFIC 
                 
                     
                 
                   PALEFTKKFSEWGNNAFFYYFEHRSSKLPWPEWMGVMHGYEIEFVFGLPL 
                 
                     
                 
                   ERRDNYTKAEEILSRSIVKRWANFAKYGNPNETQNNSTSWPVFKSTEQKY 
                 
                     
                 
                   LTLNTESTRIMTKLRAQQCRFWTSFFPKV, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         (b) a human serum albumin (HSA) polypeptide comprising the sequence 
       
       
         
           
                 
               
                   (SEQ ID NO: 2) 
                 
                   DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFA 
                 
                     
                 
                   KTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNE 
                 
                     
                 
                   CFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFY 
                 
                     
                 
                   APELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKC 
                 
                     
                 
                   ASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDL 
                 
                     
                 
                   LECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPA 
                 
                     
                 
                   DLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSVVLLLRLA 
                 
                     
                 
                   KTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGE 
                 
                     
                 
                   YKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAE 
                 
                     
                 
                   DYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPK 
                 
                     
                 
                   EFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDD 
                 
                     
                 
                   FAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and
 (c) a signal peptide comprising the sequence MRPTWAWWLFLVLLLALWAPARG (SEQ ID NO:3), 
 
       wherein the amount of the fusion protein is effective to cause attenuation of the biological effect of the cocaine exposure in the primate. 
     
     
         2 . The method of  claim 1 , wherein the fusion protein comprises the sequence 
       
         
           
                 
               
                   (SEQ ID NO: 4) 
                 
                   MRPTWAWWLFLVLLLALWAPARGEDDIIIATKNGKVRGMNLTVFGGTVTA 
                 
                     
                 
                   FLGIPYAQPPLGRLRFKKPQSLTKWSDIWNATKYANSCCQNIDQSFPGFH 
                 
                     
                 
                   GSEMWNPNTDLSEDCLYLNVWIPAPKPKNATVLIWIYGGGFQTGTSSLHV 
                 
                     
                 
                   YDGKFLARVERVIVVSMNYRVGALGFLALPGNPEAPGNMGLFDQQLALQW 
                 
                     
                 
                   VQKNIAAFGGNPKSVTLFGESSGAASVSLHLLSPGSHSLFTRAILQSGSF 
                 
                     
                 
                   NAPWAVTSLYEARNRTLNLAKLTGCSRENETEIIKCLRNKDPQEILLNEA 
                 
                     
                 
                   FVVPYGTPLGVNFGPTVDGDFLTDMPDILLELGQFKKTQILVGVNKDEGT 
                 
                     
                 
                   WFLVGGAPGFSKDNNSIITRKEFQEGLKIFFPGVSEFGKESILFHYTDWV 
                 
                     
                 
                   DDQRPENYREALGDVVGDYNFICPALEFTKKFSEWGNNAFFYYFEHRSSK 
                 
                     
                 
                   LPWPEWMGVMHGYEIEFVFGLPLERRDNYTKAEEILSRSIVKRWANFAKY 
                 
                     
                 
                   GNPNETQNNSTSWPVFKSTEQKYLTLNTESTRIMTKLRAQQCRFWTSFFP 
                 
                     
                 
                   KVDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTE 
                 
                     
                 
                   FAKTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPER 
                 
                     
                 
                   NECFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPY 
                 
                     
                 
                   FYAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRL 
                 
                     
                 
                   KCASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHG 
                 
                     
                 
                   DLLECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEM 
                 
                     
                 
                   PADLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSVVLLLR 
                 
                     
                 
                   LAKTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQL 
                 
                     
                 
                   GEYKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPC 
                 
                     
                 
                   AEDYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYV 
                 
                     
                 
                   PKEFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVM 
                 
                     
                 
                   DDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 - 45 . (canceled) 
     
     
         46 . A method of attenuating a biological effect of a cocaine exposure in a primate comprising administering to the primate an amount of a fusion protein comprising:
 (a) a mutant butyrylcholinesterase (BChE) polypeptide comprising the sequence   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   EDDIIIATKNGKVRGMNLTVFGGTVTAFLGIPYAQPPLGRLRFKKPQSLT 
                 
                     
                 
                   KWSDIWNATKYANSCCQNIDQSFPGFHGSEMWNPNTDLSEDCLYLNVWIP 
                 
                     
                 
                   APKPKNATVLIWIYGGGFQTGTSSLHVYDGKFLARVERVIVVSMNYRVGA 
                 
                     
                 
                   LGFLALPGNPEAPGNMGLFDQQLALQWVQKNIAAFGGNPKSVTLFGESSG 
                 
                     
                 
                   AASVSLHLLSPGSHSLFTRAILQSGSFNAPWAVTSLYEARNRTLNLAKLT 
                 
                     
                 
                   GCSRENETEIIKCLRNKDPQEILLNEAFVVPYGTPLGVNFGPTVDGDFLT 
                 
                     
                 
                   DMPDILLELGQFKKTQILVGVNKDEGTWFLVGGAPGFSKDNNSIITRKEF 
                 
                     
                 
                   QEGLKIFFPGVSEFGKESILFHYTDWVDDQRPENYREALGDVVGDYNFIC 
                 
                     
                 
                   PALEFTKKFSEWGNNAFFYYFEHRSSKLPWPEWMGVMHGYEIEFVFGLPL 
                 
                     
                 
                   ERRDNYTKAEEILSRSIVKRWANFAKYGNPNETQNNSTSWPVFKSTEQKY 
                 
                     
                 
                   LTLNTESTRIMTKLRAQQCRFWTSFFPKV, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and
 (b) a human serum albumin (HSA) polypeptide comprising the sequence 
 
       
         
           
                 
               
                   (SEQ ID NO: 2) 
                 
                   DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFA 
                 
                     
                 
                   KTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNE 
                 
                     
                 
                   CFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFY 
                 
                     
                 
                   APELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKC 
                 
                     
                 
                   ASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDL 
                 
                     
                 
                   LECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPA 
                 
                     
                 
                   DLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSVVLLLRLA 
                 
                     
                 
                   KTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGE 
                 
                     
                 
                   YKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAE 
                 
                     
                 
                   DYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPK 
                 
                     
                 
                   EFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDD 
                 
                     
                 
                   FAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       wherein the amount of the fusion protein is effective to cause attenuation of the biological effect of the cocaine exposure in the primate. 
     
     
         47 . The method of  claim 46 , wherein the fusion protein comprises the sequence 
       
         
           
                 
               
                   (Residues 24 to 1137 of SEQ ID NO: 4) 
                 
                   EDDIIIATKNGKVRGMNLTVFGGTVTAFLGIPYAQPPLGRLRFKKPQSLT 
                 
                     
                 
                   KWSDIWNATKYANSCCQNIDQSFPGFHGSEMWNPNTDLSEDCLYLNVWIP 
                 
                     
                 
                   APKPKNATVLIWIYGGGFQTGTSSLHVYDGKFLARVERVIVVSMNYRVGA 
                 
                     
                 
                   LGFLALPGNPEAPGNMGLFDQQLALQWVQKNIAAFGGNPKSVTLFGESSG 
                 
                     
                 
                   AASVSLHLLSPGSHSLFTRAILQSGSFNAPWAVTSLYEARNRTLNLAKLT 
                 
                     
                 
                   GCSRENETEIIKCLRNKDPQEILLNEAFVVPYGTPLGVNFGPTVDGDFLT 
                 
                     
                 
                   DMPDILLELGQFKKTQILVGVNKDEGTWFLVGGAPGFSKDNNSIITRKEF 
                 
                     
                 
                   QEGLKIFFPGVSEFGKESILFHYTDWVDDQRPENYREALGDVVGDYNFIC 
                 
                     
                 
                   PALEFTKKFSEWGNNAFFYYFEHRSSKLPWPEWMGVMHGYEIEFVFGLPL 
                 
                     
                 
                   ERRDNYTKAEEILSRSIVKRWANFAKYGNPNETQNNSTSWPVFKSTEQKY 
                 
                     
                 
                   LTLNTESTRIMTKLRAQQCRFWTSFFPKVDAHKSEVAHRFKDLGEENFKA 
                 
                     
                 
                   LVLIAFAQYLQQCPFEDHVKLVNEVTEFAKTCVADESAENCDKSLHTLFG 
                 
                     
                 
                   DKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVD 
                 
                     
                 
                   VMCTAFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQA 
                 
                     
                 
                   ADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGERAFKAWAVARLSQ 
                 
                     
                 
                   RFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSI 
                 
                     
                 
                   SSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESEDVCKNYAE 
                 
                     
                 
                   AKDVFLGMFLYEYARRHPDYSVVLLLRLAKTYETTLEKCCAAADPHECYA 
                 
                     
                 
                   KVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTP 
                 
                     
                 
                   TLVEVSRNLGKVGSKCCKHPEAKRMPCAEDYLSVVLNQLCVLHEKTPVSD 
                 
                     
                 
                   RVTKCCTESLVNRRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKER 
                 
                     
                 
                   QIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEE 
                 
                     
                 
                   GKKLVAASQAALGL. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         48 . The method of  claim 46 , wherein the fusion protein is administered by intramuscular injection. 
     
     
         49 . A pharmaceutical composition comprising:
 (I) a fusion protein comprising:   (a) a mutant butyrylcholinesterase (BChE) polypeptide comprising the sequence   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   EDDIIIATKNGKVRGMNLTVFGGTVTAFLGIPYAQPPLGRLRFKKPQSLT 
                 
                     
                 
                   KWSDIWNATKYANSCCQNIDQSFPGFHGSEMWNPNTDLSEDCLYLNVWIP 
                 
                     
                 
                   APKPKNATVLIWIYGGGFQTGTSSLHVYDGKFLARVERVIVVSMNYRVGA 
                 
                     
                 
                   LGFLALPGNPEAPGNMGLFDQQLALQWVQKNIAAFGGNPKSVTLFGESSG 
                 
                     
                 
                   AASVSLHLLSPGSHSLFTRAILQSGSFNAPWAVTSLYEARNRTLNLAKLT 
                 
                     
                 
                   GCSRENETEIIKCLRNKDPQEILLNEAFVVPYGTPLGVNFGPTVDGDFLT 
                 
                     
                 
                   DMPDILLELGQFKKTQILVGVNKDEGTWFLVGGAPGFSKDNNSIITRKEF 
                 
                     
                 
                   QEGLKIFFPGVSEFGKESILFHYTDWVDDQRPENYREALGDVVGDYNFIC 
                 
                     
                 
                   PALEFTKKFSEWGNNAFFYYFEHRSSKLPWPEWMGVMHGYEIEFVFGLPL 
                 
                     
                 
                   ERRDNYTKAEEILSRSIVKRWANFAKYGNPNETQNNSTSWPVFKSTEQKY 
                 
                     
                 
                   LTLNTESTRIMTKLRAQQCRFWTSFFPKV, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and
 (b) a human serum albumin (HSA) polypeptide comprising the sequence 
 
       
         
           
                 
               
                   (SEQ ID NO: 2) 
                 
                   DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFA 
                 
                     
                 
                   KTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNE 
                 
                     
                 
                   CFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFY 
                 
                     
                 
                   APELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKC 
                 
                     
                 
                   ASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDL 
                 
                     
                 
                   LECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPA 
                 
                     
                 
                   DLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSVVLLLRLA 
                 
                     
                 
                   KTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGE 
                 
                     
                 
                   YKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAE 
                 
                     
                 
                   DYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPK 
                 
                     
                 
                   EFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDD 
                 
                     
                 
                   FAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and
 (II) a formulation buffer comprising 10 mM sodium phosphate, 200 mM mannitol, 60 mM trehalose, and 0.01% (w/v) polysorbate 80, pH 7.2.

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