Methods to identify true antagonists and inverse agonists of the cannabinoid receptor
AssigneeAstraZeneca A B
InventorPeter Greasley
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Abstract
The present invention relates to a method to identify a true antagonist and an inverse agonist of a cannabinoid receptor (CB), and to discriminate between them. The invention further relates to the use of these true antagonists and inverse agonists in the treatment of CB associated disorders such as obesity, psychiatric and neurological disorders.
Claims31
A method for identifying an inverse agonist of a CB receptor, the method comprising:
- measuring the activity of a constitutively active CB receptor;
- contacting a CB receptor test inhibitory agent with the constitutively active CB receptor;
- and
- measuring the activity of the constitutively active CB receptor following contact with the inhibitory agent, wherein a decrease in the activity in the constitutively active CB receptor, compared to the activity of the constitutively active CB receptor in the absence of the inhibitory agent, indicates that the agent is an inverse agonist.
A method for determining if a CB receptor inhibitory agent is an inverse agonist or a true antagonist of a CB receptor, the method comprising:
- providing a test CB receptor inhibitory agent;
- contacting the agent with a wild-type CB receptor in the presence of a CB receptor agonist;
- contacting the agent with a constitutively active CB receptor;
- measuring the activity of the wild-type CB receptor and the constitutively active CB receptor, wherein:
- (i) a decrease in the activity in both the wild-type CB receptor and the constitutively active CB receptor indicates that the agent is an inverse agonist, or
- (ii) a decrease in the activity in the wild-type CB receptor, but not of the activity of the constitutively active CB receptor, indicates that the compound is a true antagonist.
A method for identifying an inverse agonist of a CB receptor, the method comprising:
- measuring the activity of a constitutively active CB receptor expressed in a cell;
- contacting a CB receptor test inhibitory agent with the cell expressing the constitutively active CB receptor;
- and
- measuring the activity of the constitutively active CB receptor following contact with the inhibitory agent, wherein a decrease in the activity in the constitutively active CB receptor compared to the activity of the constitutively active CB receptor in the absence of the inhibitory agent indicates that the agent is an inverse agonist.
A method for determining if a CB receptor inhibitory agent is an inverse agonist or a true antagonist of a CB receptor, the method comprising:
- providing a test CB receptor inhibitory agent;
- contacting the agent with a cell expressing a wild-type CB receptor in the presence of a CB agonist;
- contacting the agent with a cell expressing a constitutively active CB receptor;
- measuring the activity of the wild-type CB receptor and the constitutively active CB receptor, wherein
- (i) a decrease in the activity in both the wild-type CB receptor and the constitutively active CB receptor indicates that the agent is an inverse agonist;
- or
- (ii) a decrease in the activity in the wild-type CB receptor in the presence of a CB receptor agonist, but not the activity of the constitutively active CB receptor, indicates that the compound is a true antagonist
(canceled)
(canceled)
A constitutively active CB receptor.
An isolated nucleic acid sequence comprising a nucleotide sequence that encodes a variant cannabinoid receptor protein wherein one or both of the amino acids located at positions 3:49 and 6:32 have been substituted for by another amino acid so as to create a constitutive vaiant form of the cannabinoid receptor.
An isolated cannabinoid receptor polypeptide, wherein one or both of the natural amino acids at positions 3:49 and 6:32 of the receptor polypeptide have been substituted for another amino acid.
An isolated human cannabinoid 1 receptor polypeptide comprising the sequence according to SEQ ID NO:
- 1, with the exception that one or both of the amino acids at position 213 or 338 may be an aspartic acid residue.
Dependencies are inferred from the claim text.