Agonist inverse agonist characterization at CB1 and CB2 cannabinoid receptors of L759633, L759656 and AM630

R A Ross, H C Brockie, L A Stevenson, V L Murphy, F Templeton, A Makriyannis, R G Pertwee

Research output: Contribution to journalArticle

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Abstract

1 We have tested our prediction that AM630 is a CB2 cannabinoid receptor ligand and also investigated whether L759633 and L759656, are CB2 receptor agonists.

2 Binding assays with membranes from CHO cells stably transfected with human CB1 or CB2 receptors using [H-3]-CP55940, confirmed the CB2-selectivity of L759633 and L759656 (CB2/CB1 affinity ratios=163 and 414 respectively) and showed AM630 to have a K-1 at CB2 receptors of 31.2 nM and a CB2/CB1 affinity ratio of 165.

3 In CB2-transfected cells, L759633 and L759656 were potent inhibitors of forskolin-stimulated cyclic AMP production, with EC50 values of 8.1 and 3.1 nM respectively and CB1/CB2 EC50 ratios of >1000 and >3000 respectively.

4 AM630 inhibited [S-35]-GTP gamma S binding to CB2 receptor membranes (EC50 = 76.6 nM), enhanced forskolin-stimulated cyclic AMP production in CB2-transfected cells (5.2 fold by 1 mu M), and antagonized the inhibition of forskolin-stimulated cyclic AMP production in this cell line induced by CP55940.

5 In CB1-transfected cells, forskolin-stimulated cyclic AMP production was significantly inhibited by AM630 (22.6% at 1 mu M and 45.9% at 10 mu M) and by L759633 at 10 mu M (48%) but not 1 mu M. L759656 (10 mu M) was not inhibitory. AM630 also produced a slight decrease in the mean inhibitory effect of CP55940 on cyclic AMP production which was not statistically significant.

6 We conclude that AM630 is a CB2-selective ligand that behaves as an inverse agonist at CB2 receptors and as a weak partial agonist at CB1 receptors. L759633 and L759656 are both potent CB2-selective agonists.

Original languageEnglish
Pages (from-to)665-672
Number of pages8
JournalBritish Journal of Pharmacology
Volume126
Issue number3
DOIs
Publication statusPublished - Feb 1999

Keywords

  • cannabinoid CB1 and CB2 receptors
  • AM630
  • L759633
  • L759656
  • CB2-selective ligands
  • CB2-selective agonist
  • CB2-selective inverse agonist
  • [S-35]-GTP gamma S
  • cyclic AMP
  • BINDING
  • BRAIN
  • PHARMACOLOGY
  • ANTAGONIST

Cite this

Agonist inverse agonist characterization at CB1 and CB2 cannabinoid receptors of L759633, L759656 and AM630. / Ross, R A ; Brockie, H C ; Stevenson, L A ; Murphy, V L ; Templeton, F ; Makriyannis, A ; Pertwee, R G .

In: British Journal of Pharmacology, Vol. 126, No. 3, 02.1999, p. 665-672.

Research output: Contribution to journalArticle

Ross, R A ; Brockie, H C ; Stevenson, L A ; Murphy, V L ; Templeton, F ; Makriyannis, A ; Pertwee, R G . / Agonist inverse agonist characterization at CB1 and CB2 cannabinoid receptors of L759633, L759656 and AM630. In: British Journal of Pharmacology. 1999 ; Vol. 126, No. 3. pp. 665-672.
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abstract = "1 We have tested our prediction that AM630 is a CB2 cannabinoid receptor ligand and also investigated whether L759633 and L759656, are CB2 receptor agonists.2 Binding assays with membranes from CHO cells stably transfected with human CB1 or CB2 receptors using [H-3]-CP55940, confirmed the CB2-selectivity of L759633 and L759656 (CB2/CB1 affinity ratios=163 and 414 respectively) and showed AM630 to have a K-1 at CB2 receptors of 31.2 nM and a CB2/CB1 affinity ratio of 165.3 In CB2-transfected cells, L759633 and L759656 were potent inhibitors of forskolin-stimulated cyclic AMP production, with EC50 values of 8.1 and 3.1 nM respectively and CB1/CB2 EC50 ratios of >1000 and >3000 respectively.4 AM630 inhibited [S-35]-GTP gamma S binding to CB2 receptor membranes (EC50 = 76.6 nM), enhanced forskolin-stimulated cyclic AMP production in CB2-transfected cells (5.2 fold by 1 mu M), and antagonized the inhibition of forskolin-stimulated cyclic AMP production in this cell line induced by CP55940.5 In CB1-transfected cells, forskolin-stimulated cyclic AMP production was significantly inhibited by AM630 (22.6{\%} at 1 mu M and 45.9{\%} at 10 mu M) and by L759633 at 10 mu M (48{\%}) but not 1 mu M. L759656 (10 mu M) was not inhibitory. AM630 also produced a slight decrease in the mean inhibitory effect of CP55940 on cyclic AMP production which was not statistically significant.6 We conclude that AM630 is a CB2-selective ligand that behaves as an inverse agonist at CB2 receptors and as a weak partial agonist at CB1 receptors. L759633 and L759656 are both potent CB2-selective agonists.",
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T1 - Agonist inverse agonist characterization at CB1 and CB2 cannabinoid receptors of L759633, L759656 and AM630

AU - Ross, R A

AU - Brockie, H C

AU - Stevenson, L A

AU - Murphy, V L

AU - Templeton, F

AU - Makriyannis, A

AU - Pertwee, R G

PY - 1999/2

Y1 - 1999/2

N2 - 1 We have tested our prediction that AM630 is a CB2 cannabinoid receptor ligand and also investigated whether L759633 and L759656, are CB2 receptor agonists.2 Binding assays with membranes from CHO cells stably transfected with human CB1 or CB2 receptors using [H-3]-CP55940, confirmed the CB2-selectivity of L759633 and L759656 (CB2/CB1 affinity ratios=163 and 414 respectively) and showed AM630 to have a K-1 at CB2 receptors of 31.2 nM and a CB2/CB1 affinity ratio of 165.3 In CB2-transfected cells, L759633 and L759656 were potent inhibitors of forskolin-stimulated cyclic AMP production, with EC50 values of 8.1 and 3.1 nM respectively and CB1/CB2 EC50 ratios of >1000 and >3000 respectively.4 AM630 inhibited [S-35]-GTP gamma S binding to CB2 receptor membranes (EC50 = 76.6 nM), enhanced forskolin-stimulated cyclic AMP production in CB2-transfected cells (5.2 fold by 1 mu M), and antagonized the inhibition of forskolin-stimulated cyclic AMP production in this cell line induced by CP55940.5 In CB1-transfected cells, forskolin-stimulated cyclic AMP production was significantly inhibited by AM630 (22.6% at 1 mu M and 45.9% at 10 mu M) and by L759633 at 10 mu M (48%) but not 1 mu M. L759656 (10 mu M) was not inhibitory. AM630 also produced a slight decrease in the mean inhibitory effect of CP55940 on cyclic AMP production which was not statistically significant.6 We conclude that AM630 is a CB2-selective ligand that behaves as an inverse agonist at CB2 receptors and as a weak partial agonist at CB1 receptors. L759633 and L759656 are both potent CB2-selective agonists.

AB - 1 We have tested our prediction that AM630 is a CB2 cannabinoid receptor ligand and also investigated whether L759633 and L759656, are CB2 receptor agonists.2 Binding assays with membranes from CHO cells stably transfected with human CB1 or CB2 receptors using [H-3]-CP55940, confirmed the CB2-selectivity of L759633 and L759656 (CB2/CB1 affinity ratios=163 and 414 respectively) and showed AM630 to have a K-1 at CB2 receptors of 31.2 nM and a CB2/CB1 affinity ratio of 165.3 In CB2-transfected cells, L759633 and L759656 were potent inhibitors of forskolin-stimulated cyclic AMP production, with EC50 values of 8.1 and 3.1 nM respectively and CB1/CB2 EC50 ratios of >1000 and >3000 respectively.4 AM630 inhibited [S-35]-GTP gamma S binding to CB2 receptor membranes (EC50 = 76.6 nM), enhanced forskolin-stimulated cyclic AMP production in CB2-transfected cells (5.2 fold by 1 mu M), and antagonized the inhibition of forskolin-stimulated cyclic AMP production in this cell line induced by CP55940.5 In CB1-transfected cells, forskolin-stimulated cyclic AMP production was significantly inhibited by AM630 (22.6% at 1 mu M and 45.9% at 10 mu M) and by L759633 at 10 mu M (48%) but not 1 mu M. L759656 (10 mu M) was not inhibitory. AM630 also produced a slight decrease in the mean inhibitory effect of CP55940 on cyclic AMP production which was not statistically significant.6 We conclude that AM630 is a CB2-selective ligand that behaves as an inverse agonist at CB2 receptors and as a weak partial agonist at CB1 receptors. L759633 and L759656 are both potent CB2-selective agonists.

KW - cannabinoid CB1 and CB2 receptors

KW - AM630

KW - L759633

KW - L759656

KW - CB2-selective ligands

KW - CB2-selective agonist

KW - CB2-selective inverse agonist

KW - [S-35]-GTP gamma S

KW - cyclic AMP

KW - BINDING

KW - BRAIN

KW - PHARMACOLOGY

KW - ANTAGONIST

U2 - 10.1038/sj.bjp.0702351

DO - 10.1038/sj.bjp.0702351

M3 - Article

VL - 126

SP - 665

EP - 672

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 3

ER -