Bivalent binding molecules of 7 transmembrane G protein-coupled receptors

Abstract

Described herein are methods for identifying and preparing bivalent binding molecules to 7 transmembrane G protein-coupled receptors. The methods disclosed herein are based on the SELEX method for generating high affinity nucleic acid ligands. SELEX is an acronym for Systematic Evolution of Ligands by EXponential enrichment. The methods of this invention combine two or more binding domains to two or more different epitopes of the same 7 transmembrane G protein-coupled receptor. These bivalent binding molecules are useful as therapeutic and diagnostic agents.

Description

RELATEDNESS OF THE APPLICATION [0001] The subject application is a divisional application of co-pending U.S. Ser. No. 10 / 729,667, filed Dec. 5, 2003, which is a divisional application of U.S. Ser. No. 09 / 118,525, filed Jul. 17, 1998, now U.S. patent Ser. No. 09 / 118,525; which is a continuation-in-part of U.S. Ser. No. 08 / 956,699, filed Oct. 23, 1997, now U.S. Pat. No. 6,083,696; which is a continuation of U.S. Ser. No. 08 / 234,997, filed Apr. 28, 1994, now U.S. Pat. No. 5,683,867; each of which are hereby incorporated in their entirety by reference.FIELD OF THE INVENTION [0002] Described herein are bivalent binding molecules that can activate or inhibit 7 transmembrane G protein-coupled receptors. Also described herein are methods for identifying and preparing bivalent binding molecules to 7 transmembrane G protein-coupled receptors. The methods disclosed herein are based on the SELEX method for generating high affinity nucleic acid ligands, also termed aptamers. SELEX is an acronym ...

AI Technical Summary

Benefits of technology

[0113] The amount of the bivalent compound which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, which can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness or advancement of the disease or condition, and should be decided according to the practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curved derived from in vitro or animal model test systems. For example, an effective amount of a bivalent compound of the invention is readily determined by administering graded doses of a bivalent compound of the invention and observing the desired effect.

[0114] The following examples are meant to illustrate the method of the invention and are not intended to limit the scope, utility, or applicability of this invention.

Problems solved by technology

However, these receptors are difficult to purify.

In addition, most membrane proteins are not soluble in water.

Attempts to achieve expression of only the ligand binding portion of a 7TM G protein-coupled receptor have been unreproducible or have resulted in inefficient and / or unpredictable levels of expression (Xie, U. B., et al, J. Biol. Chem. 265:21441-21420 (1990); Tsai-Morris, C. H., et al.

Small peptides have flexible structures and usually exist in solution as an equilibrium of multiple conformers, and thus it was initially thought that binding affinities may be limited by the conformational entropy lost upon binding a flexible peptide.

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