Patent ID: 6406866
Filing Date: 2002-06-18
Classification: A61K,C07K,C12N

Abstract:
A method of screening a test compound for a capacity to competitively bind to a mammalian methadone-specific opioid receptor (MSOR) in cells expressing the MSOR, the method comprising:(a) transforming a host cell with an expression vector comprising a nucleic acid encoding the MSOR; (b) culturing the transformed host cell under conditions that would allow expression of the MSOR; (c) assaying the transformed cell culture for binding of an amount of a detectably-labeled peptide, wherein the peptide has a sequence comprising: Xaa-Gly-Gly-Phe-(A1)-(A2)-(A3)-(A4)-(A5)-(A6)-(A7)-(A8)-(A9)-(A10)-(A11)-(A12)-Gln wherein Xaa is Phe or Tyr; A1 is Tbr, Leu or Met; A2 is Gly, Arg or Thr; A3 is Ala, Arg or Ser; A4 is Arg, Ile, Glu or Gln; A5 is Lys, Arg, or Phe; A6 is Ser, Pro or Lys; A7 is Ala, Lys, Gln or Val; A8 is Arg, Leu, Thr or Val; A9 is Lys, Pro or Thr; A10 is Tyr, Leu or Trp; A11 is Ala, Asp or Val; A12 is Asn or Thr; (SEQ ID NO:15), wherein at least one amino acid at position A1 to A12 is uniquely found in a corresponding position of SEQ ID NO:5 or 6; and wherein amino acids are each individually in either the D or L stereochemical configuration and wherein the peptide specifically binds to a mammalian MSOR in competition with varying amounts of the test compound; and (d) determining whether the test compound competitively binds to the MSOR by calculating an extent of inhibition of binding of the detectably-labeled peptide in the presence of the test compound.