Skip to main content
Free research Free research Free assay review with research home and workspace preview
Quick search ChEMBL 36
Assay Detail

CHEMBL5736764

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
Determination of Binding Affinity for CGRP Receptor by Radioligand Binding Assay in SK-N-MC Cell Membranes: The binding affinity assay of compounds for human CGRP receptor was carried out by inhibition of radiolabeled ligand [125I]-CGRP binding in human neuroblastoma cell line SK-N-MC cell membranes.Cell membranes prepared from SK-N-MC cells expressing CGRP receptors endogenously (Muff et al., Ann N Y Acad Sci. 1992; 657: 106-116) were used for radioligand binding assay. Radioligand binding assay was performed using 96-well microplate in a total volume of 200 μL in each well. A 2.5 μL of serial dilution of compound dissolved in dimethylsulfoxide (DMSO) was mixed with SK-N-MC cell membranes (40 μg membrane protein per well) and [125I]-CGRP (PerkinElmer NEX354, final concentration of 150 pM) in an assay buffer consisting of 50 mM Tris-HCl, 5 mM MgCl2 and 0.1% bovine serum albumin (pH 7.4). The assay plate was incubated with shaking on a plate shaker at room temperature for 90 minutes. The incubation was terminated by filtration through a GF/C glass fiber filter plate (Merck Millipore) pre-soaked with 0.3% polyethyleneimine (PEI). Filters were washed 4 times with 300 μL of ice-cold assay buffer. After drying the filter plate, 100 μL of scintillation fluid (PerkinElmer, MicroScint-20) was added to each well and the radioactivity was counted using a TopCount NXT (PerkinElmer). Non-specific binding was determined in the presence of 1.25 μM unlabelled human α-CGRP (Bachem). The radioactivity was converted to the percent of specific binding using the equation below.% ⁢ ⁢ of ⁢ ⁢ specific ⁢ ⁢ binding = ( Y - Y ⁢ ⁢ min ) ( Y ⁢ ⁢ max - Y ⁢ ⁢ min ) × 100 [ Number ⁢ ⁢ 1 ]In the formulae, Y is observed radioactivity, Ymax is total bound activity, and Ymin is non-specific bound activity.From these data, the concentration of compound required for 50% inhibition of radioligand binding (IC50) was determined using Prism (GraphPad Inc.). The IC50 value is then converted to the equilibrium dissociation constant (Ki) using the Cheng-Prusoff equation below (Cheng & Prusoff (1973) Biochem. Pharmacol. 22, 3099-3108).Ki = IC 50 1 + [ L ] Kd [ Number ⁢ ⁢ 2 ]In the formulae, [L] is the concentration of radioligand, Kd is the apparent dissociation constant of the radioligand for the receptor as determined by saturation binding assay with [125I]-CGRP.
ChEMBL ↗
576
Total Activities
116
Compounds Tested
4
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

Calcitonin gene-related peptide type 1 receptor (CHEMBL3798) ↗
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Pyrrolidine derivative
(2021)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
kon 192 - -
k_off 192 - -
Ki 116 9.80 10.70
IC50 76 10.25 11.00

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5865405 — — 6 11.00
CHEMBL6015508 — — 6 11.00
CHEMBL5755575 — — 6 11.00
CHEMBL5978732 — — 6 11.00
CHEMBL5810695 — — 6 11.00
CHEMBL5814140 — — 6 10.70
CHEMBL5988962 — — 6 10.70
CHEMBL5890201 — — 6 10.70
CHEMBL5751888 — — 6 10.70
CHEMBL5981859 — — 6 10.70
CHEMBL6052539 — — 6 10.70
CHEMBL5759145 — — 6 10.70
CHEMBL5828757 — — 6 10.70
CHEMBL5770083 — — 6 10.70
CHEMBL5760510 — — 6 10.70
CHEMBL5965987 — — 6 10.70
CHEMBL5742096 — — 6 10.70
CHEMBL5917191 — — 6 10.70
CHEMBL5740788 — — 6 10.70
CHEMBL6061451 — — 6 10.70
CHEMBL5819831 — — 6 10.52
CHEMBL5980403 — — 6 10.52
CHEMBL5939455 — — 6 10.52
CHEMBL5962492 — — 6 10.52
CHEMBL5800111 — — 6 10.52
CHEMBL6032212 — — 6 10.52
CHEMBL5854158 — — 6 10.52
CHEMBL5824600 — — 6 10.52
CHEMBL5937861 — — 6 10.52
CHEMBL5838667 — — 6 10.52

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5810695 — IC50 = 0.01 nM 11.00
CHEMBL6015508 — IC50 = 0.01 nM 11.00
CHEMBL5755575 — IC50 = 0.01 nM 11.00
CHEMBL5978732 — IC50 = 0.01 nM 11.00
CHEMBL5865405 — IC50 = 0.01 nM 11.00
CHEMBL5740788 — IC50 = 0.02 nM 10.70
CHEMBL5760510 — IC50 = 0.02 nM 10.70
CHEMBL5759145 — IC50 = 0.02 nM 10.70
CHEMBL5965987 — IC50 = 0.02 nM 10.70
CHEMBL5742096 — IC50 = 0.02 nM 10.70
CHEMBL5988962 — IC50 = 0.02 nM 10.70
CHEMBL5917191 — IC50 = 0.02 nM 10.70
CHEMBL5770083 — IC50 = 0.02 nM 10.70
CHEMBL5981859 — IC50 = 0.02 nM 10.70
CHEMBL5890201 — IC50 = 0.02 nM 10.70
CHEMBL5751888 — IC50 = 0.02 nM 10.70
CHEMBL6061451 — IC50 = 0.02 nM 10.70
CHEMBL6052539 — IC50 = 0.02 nM 10.70
CHEMBL5828757 — Ki = 0.02 nM 10.70
CHEMBL5814140 — IC50 = 0.02 nM 10.70
CHEMBL5937861 — IC50 = 0.03 nM 10.52
CHEMBL5819831 — IC50 = 0.03 nM 10.52
CHEMBL5854158 — IC50 = 0.03 nM 10.52
CHEMBL5934958 — IC50 = 0.03 nM 10.52
CHEMBL5824600 — IC50 = 0.03 nM 10.52
CHEMBL5980403 — IC50 = 0.03 nM 10.52
CHEMBL6032212 — IC50 = 0.03 nM 10.52
CHEMBL5800111 — IC50 = 0.03 nM 10.52
CHEMBL5962492 — IC50 = 0.03 nM 10.52
CHEMBL6062910 — IC50 = 0.03 nM 10.52
CHEMBL5939455 — IC50 = 0.03 nM 10.52
CHEMBL5838667 — Ki = 0.03 nM 10.52
CHEMBL6011978 — IC50 = 0.03 nM 10.52
CHEMBL5781964 — IC50 = 0.03 nM 10.52
CHEMBL5980403 — Ki = 0.04 nM 10.40
CHEMBL5773633 — IC50 = 0.04 nM 10.40
CHEMBL5820122 — IC50 = 0.04 nM 10.40
CHEMBL5996912 — IC50 = 0.04 nM 10.40
CHEMBL5850727 — IC50 = 0.04 nM 10.40
CHEMBL5838667 — IC50 = 0.04 nM 10.40
CHEMBL5824600 — Ki = 0.04 nM 10.40
CHEMBL6020593 — IC50 = 0.04 nM 10.40
CHEMBL5918805 — IC50 = 0.04 nM 10.40
CHEMBL5797733 — IC50 = 0.04 nM 10.40
CHEMBL6001404 — IC50 = 0.04 nM 10.40
CHEMBL5918804 — IC50 = 0.04 nM 10.40
CHEMBL5820122 — Ki = 0.04 nM 10.40
CHEMBL5996912 — Ki = 0.05 nM 10.30
CHEMBL5745025 — IC50 = 0.05 nM 10.30
CHEMBL5760510 — Ki = 0.05 nM 10.30