Screening methods using g-protein coupled receptors and related compositions
AssigneesThe General Hospital Corporation, Chugai Pharmaceutical Co., Ltd.
InventorsThomas J. Gardella, John T. Potts, JR., Masaru Shimizu, Fumihiko Ichikawa, Harald Juppner, Makoto Okazaki
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Abstract
The present invention provides screening methods for GPCRs based on the discovery that the affinity of a receptor agonist for a GPCR (such as the parathyroid hormone receptor) when not bound to a G-protein is correlated with the length of time over which the agonist is effective, independently of its pharmacokinetic properties. The invention also provides PTH- and PTHrP-derived polypeptides.
Claims71
A method for determining whether a candidate compound is a long-acting agonist of a G protein coupled receptor (GPCR), said method comprising:
- (a) contacting said GPCR with said compound, wherein said GPCR is in the RG form;
- (b) measuring the affinity of said compound for the RG form of said GPCR;
- (c) contacting said GPCR with said compound, wherein said GPCR is in the R0 form;
- and
- (d) measuring the affinity of said compound for the R0 form of said GPCR, wherein a compound that (i) that has an affinity for the RG form of said GPCR that is at least 10% of an endogenous agonist for said GPCR, and (ii) has a greater affinity for the R0 form of said GPCR than said endogenous agonist is identified as a long-acting agonist of said GPCR.
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A method for determining whether a candidate compound is a short-acting agonist of a G protein coupled receptor (GPCR), said method comprising:
- (a) contacting said GPCR with said compound, wherein said GPCR is in the RG form;
- (b) measuring the affinity of said compound for the RG form of said GPCR;
- (c) contacting said GPCR with said compound, wherein said GPCR is in the R0 form;
- and
- (d) measuring the affinity of said compound for the R0 form of said GPCR, wherein a compound that (i) that has an affinity for the RG form of said GPCR that is at least 10% of an endogenous agonist for said GPCR, and (ii) has a lower affinity for the R0 form of said GPCR than said endogenous agonist is identified as a short-acting agonist of said GPCR.
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A polypeptide having affinity for PTH RG and a low affinity for PTH R0.
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A polypeptide which binds the PTH receptor and has a high affinity for the R0 form of the PTH receptor.
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A polypeptide comprising the amino acid sequence:
- (SEQ ID NO:
- 33)
- Ala-Val-Ser-Glu-His-Glu-Leu-Leu-His-Asp-Lys-Gly-
- Lys-Ser-Ile-Gln-Asp-X1-Arg-Arg-Arg-X2-Phe-Leu-
- X3-X4-Leu-Ile-X5-X6-X7-X8-X9-X10-Glu-Ile
- wherein:
- X1 is Leu, Ala, Ser, Met, Phe, or Glu;
- X2 is Phe, Ala, Ser, Leu, Asn, Trp, Glu, or Lys;
- X3 is His, Leu, Arg, Lys, Trp, Ile, or Phe;
- X4 is His, Ala, Ser, Asn, Lys, or Arg;
- X5 is Ala, Gly, Ser, Asn, Gln, Trp, Glu, or Lys;
- X6 is Glu, Gly, Ser, Leu, Asn, Asp, Lys, or Ala;
- X7 is Ile, Leu, Val, Lys, or Ala;
- X8 is His or Ala
- X9 is Thr, Asn, or Ala;
- and
- X10 is Ala or Phe,
- or a fragment thereof containing amino acids 1-24, 1-25, 1-26, 1-27, 1-28, 1-29, 1-30, 1-31, 1-32, 1-33, 1-34, or 1-35 of said amino acid sequence, and wherein said polypeptide comprises at least one amino acid substitution as compared to the corresponding wild type PTHrP sequence or a fragment thereof;
- or a pharmaceutically acceptable salt thereof.
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Dependencies are inferred from the claim text.