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Methods for identifying compounds for regulating muscle mass or function using dopamine receptors

A technology for muscle mass and dopamine, which can be used in the screening of compounds, chemical instruments and methods, active ingredients of heterocyclic compounds, etc., and can solve problems such as lack of screening methods

Inactive Publication Date: 2007-12-05
PROCTER & GAMBLE CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] A problem associated with identifying compounds for the treatment of skeletal muscle atrophy or muscular dystrophy is the lack of good screening methods for identifying these compounds

Method used

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  • Methods for identifying compounds for regulating muscle mass or function using dopamine receptors
  • Methods for identifying compounds for regulating muscle mass or function using dopamine receptors
  • Methods for identifying compounds for regulating muscle mass or function using dopamine receptors

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0085] IV. Preparation of dopamine receptor antibody

[0086] Antibodies that selectively recognize one or more epitopes of dopamine receptors are also encompassed by the invention. These antibodies include, for example, polyclonal antibodies, monoclonal antibodies, chimeric antibodies, human antibodies, single chain antibodies, Fab fragments, F(ab') 2 Fragments, molecules produced using Fab expression libraries, human antibodies (polyclonal or monoclonal) produced in genetically transcribed mice, and epitope-binding fragments of any of the above. For therapeutic use, chimeric or human antibodies are preferred; human antibodies are most preferred.

[0087] Antibodies can be used in conjunction with the compound screening protocols described herein to evaluate test compounds, e.g., immobilized dopamine receptor polypeptides, or these antibodies can be used in conjunction with gene therapy techniques, to evaluate, e.g., dopamine receptor expression in cells or Expression dir...

Embodiment 1

[0137] Example 1. Construction of human D 1 and D 5 Dopamine receptor expression vector.

[0138] Person D 1 and D 5 Dopamine receptors (hD 1 and hD 5dopamine receptor) DNA sequences, designation numbers X58987 and X58454, were retrieved, and two oligonucleotides, including one containing the 5' end of the gene, started at the start codon (5 'oligonucleotide), and an oligonucleotide containing a stop codon, comprising the 3' end of the gene (3' oligonucleotide), was synthesized. These oligonucleotides are designed to contain restriction endonuclease sites that are not present in the D 1 or D 5 In the dopamine receptor gene, one unique site is in the 5' oligonucleotide and a different unique restriction endonuclease site is in the 3' oligonucleotide. In addition, the 3' oligonucleotide contains a polyadenine appended signal sequence. The double-stranded cDNA of human skeletal muscle was purchased from "Universal QUICK-Clone cDNA collection" (the Universal QUICK-Clone c...

Embodiment 2

[0140] Example 2. Receptor Binding Identification

[0141] Receptor binding assay of compounds in whole cell format by placing HEK293 / CRE-LUC / pIRESneo / D from Example 1 1 or D 5 Dopamine receptor cells were grown in a 96-well polylysine-coated plate. Under 5% carbon dioxide / 95% air at 37°C, cells were seeded in DMEM medium containing 10% fetal bovine serum, penicillin / streptomycin solution, L-glutamine, and non-essential amino acids, and then Cells were cultured overnight. The medium was removed and an appropriate amount of 3H-SCH23390 in MEM (Life Technologies, Rockville, MD) + 10% Seablock (Clonetech Inc., Palo Alto, CA, USA) was added. The cells were incubated with 3H-SCH23390 for 90 minutes at room temperature, and then washed 4 times with magnesium- and calcium-free phosphate-buffered saline (Life Technologies, Rockville, MD). After the final wash, cytoscint es scintillation fluid (ICNBiomedical, Inc., Costa Mesa, CA) was added, and the plate was read on a TopCount N...

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Abstract

The present invention discloses a filtering method of identifying compound which is used to or activate (D1 or D5 dopamine receptor, sole or combined) or regulate or potentially regulate skeletal muscle quality or function in human body. The present invention also discloses a filtering method of identifying compound which is used to prolong or increase D1 or D5 dopamine receptor or its signal transduction pathways and to increase D1 or D5 dopamine receptor expression. The present invention describes a drug compound containing D1 or D5 dopamine receptor agonist, and D1 or D5 dopamine receptor antibody, and a method for enhancing skeletal muscle quality or function, or for treating skeletal muscle atrophy using D1 or D5 dopamine receptor as intervention target, as well as a method for treating skeletal muscle atrophy.

Description

technical field [0001] The present invention relates to methods of identifying candidate compounds for use in modulating skeletal muscle mass or function, or modulating the activity or expression of dopamine receptors. The invention also relates to methods of treating skeletal muscle atrophy, or using D 1 or D 5 Dopamine receptors as targets for intervention to induce skeletal muscle hypertrophy, and the use of D 1 or D 5 Dopamine receptors as targets for intervention to treat muscular dystrophy. Background of the invention [0002] dopamine receptor [0003] Dopamine has multiple physiological effects, both central and peripheral. Dopamine controls multiple functions in the brain, including motor behavior, cognition, emotion, positive reinforcement, feeding, and endocrine regulation. In the periphery dopamine acts as a regulator of cardiovascular activity (including heart and vascular function), catecholamine release, hormone secretion, renal function and gastrointes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/566G01N33/567A61K49/00A61K31/55C07K16/28G01N33/68G01N33/94
CPCG01N33/9413C07K2317/21C07K16/28G01N2800/2885G01N2500/00G01N2800/2878G01N33/6887A61K31/55A61P21/00
Inventor R·J·艾斯福特R·J·谢尔登
Owner PROCTER & GAMBLE CO
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