Journal: Cephalalgia

Article Title: Migraine therapeutics differentially modulate the CGRP pathway

doi: 10.1177/0333102420983282

Figure Lengend Snippet: Binding and signaling differences of CGRP pathway therapeutics at the CGRP receptor. (a) Flow cytometry surface binding assay shows that erenumab binds to ∼98% of human CGRP receptor (CLR/RAMP1) transiently transfected HEK293S cells while fremanezumab shows no binding to cells. Representative flow cytometry dot plots are shown from at least four independent experiments. (b) Antibody concentration-response curves from flow cytometry binding experiments plotted as a percentage of the maximal binding to SK-N-MC cells (expresses endogenous human CGRP receptor) in the absence and presence of hαCGRP (100 nM). The binding responses of isotype and fremanezumab (in the absence and presence of CGRP) are overlying. Data points represent the mean ± SD (n = 3). Comparison of fits for the erenumab binding curves in the absence and presence of CGRP showed that the shift was significantly different (**** p < 0.0001). (c) Fremanezumab, erenumab and telcagepant antagonize hαCGRP-induced cAMP signaling in SK-N-MC cells. The binding curves of isotype and DMSO are overlying. Unlike fremanezumab which has no effect, erenumab and telcagepant antagonize human adrenomedullin-induced (d) and human intermedin-induced cAMP signaling (e) in SK-N-MC cells. The responses of fremanezumab, isotype and DMSO are overlying in (d) and (e). Data points represent the mean ± SEM (n = 4).

Article Snippet: Human αCGRP, amylin, adrenomedullin, intermedin and calcitonin were purchased from Bachem.

Techniques: Binding Assay, Flow Cytometry, Transfection, Concentration Assay, Comparison

Journal:

Article Title: A comparison of the actions of BIBN4096BS and CGRP 8–37 on CGRP and adrenomedullin receptors expressed on SK-N-MC, L6, Col 29 and Rat 2 cells

doi: 10.1038/sj.bjp.0704844

Figure Lengend Snippet: Effects of BIBN4096BS on the stimulation of cAMP production by human αCGRP in the presence of the indicated concentrations of BIBN4096BS in (a) L6 cells, (b) SK-N-MC cells and (c) Col 29 cells. Data represent means±s.e.mean of two to five experiments. Points were measured in duplicate in each experiment. Data are expressed as percentage of maximum cAMP production, estimated by fitting each line to a logistic Hill equation as described in the Methods. Maximum cAMP values were as follows: L6 cells, 270±30 pmol per 106 cells; SK-N-MC cells, 240±20 pmol per 106 cells; Col 29 cells, 170±19 pmol per 106 cells. Basal values were all below 10 pmol per 106 cells.

Article Snippet: BIBN4096BS was synthesized at Boehringer Ingelheim, Pharma KG. Human αCGRP was purchased from Neosystems (Strassbourg, France).

Techniques:

Journal:

Article Title: A comparison of the actions of BIBN4096BS and CGRP 8–37 on CGRP and adrenomedullin receptors expressed on SK-N-MC, L6, Col 29 and Rat 2 cells

doi: 10.1038/sj.bjp.0704844

Figure Lengend Snippet: Effects of CGRP8–37 on the stimulation of cAMP production by human αCGRP in the presence of the indicated concentrations of antagonist in (a) L6 cells, (b) SK-N-MC cells and (c) Col 29 cells. Data represent means±s.e.mean of four to seven experiments. Points were measured in duplicate in each experiment. Data are expressed as percentage of maximum cAMP production, estimated by fitting each line to a logistic Hill equation as described in the Methods. Maximum cAMP values were as follows: L6 cells, 250±27 pmol per 106 cells; SK-N-MC cells, 230±18 pmol per 106 cells; Col 29 cells, 180±17 pmol per 106 cells. Basal values were all below 10 pmol per 106 cells.

Article Snippet: BIBN4096BS was synthesized at Boehringer Ingelheim, Pharma KG. Human αCGRP was purchased from Neosystems (Strassbourg, France).

Techniques: