Fremanezumab blocks CGRP induced dilatation in human cerebral, middle meningeal and abdominal arteries

Abstract

Background: Fremanezumab (TEV-48125) is a fully humanized anti-calcitonin gene-related peptide (CGRP) monoclonal antibody (mAb) that has shown positive results in the prevention of episodic migraine and chronic migraine. Previous preclinical studies have revealed CGRP antagonistic effects on intracranial arteries (ICA). The aim of the study was to evaluate the in vitro antagonistic effects of fremanezumab on human arteries.

Methods: Arteries were removed in conjunction with neurosurgery (cerebral, CA, and middle meningeal artery, MMA, n = 7) or reconstructive abdominal surgery (abdominal artery, AA, n = 6). Ring segments of the vessels were mounted in a sensitive myograph, the functional responses of vasoactive intestinal peptide (VIP), substance P and CGRP in increasing concentrations (10^{- 10}-10^{- 7} M) were studied using pre-contraction with 30 mM potassium chloride (KCl). The concentrations of fremanezumab or isotype control antibody (66.7 nM, 0.33 μM, 0.67 μM) were given 30 min prior to CGRP administration.

Results: All included arteries responded with a strong stable contraction to the application of 30 mM KCl. During this pre-contraction, CGRP caused a concentration-dependent relaxation which differed slightly in maximum effect (I_max) between the types of arteries (ICA = 100%; AA 80%). Fremanezumab (66.7 nM) showed a shift in the IC₅₀ value of CGRP, but no significant change in I_max. At higher doses there was also a reduction of I_max. For AA, the I_max decreased from 71% at 66.7 nM, to 4.5% with 0.33 μM of fremanezumab. Isotype control antibody did not modify the responses. There was no effect on concentration-dependent relaxation with VIP with 66.7 nM of fremanezumab or isotype control.

Conclusion: CGRP relaxes pre-contracted human arteries by 80-100%, but with different IC₅₀; the potency range was ICA < AA. The antagonistic effect and potency of fremanezumab was similar, suggesting that there are vasodilatory CGRP receptors present in all studied arteries and that the antibody may have effect in all studied vessels.

Keywords: Antibody; CGRP; CGRP receptor antagonist; Fremanezumab; Human vessels.

Fig. 1

Human abdominal subcutaneous arteries were pre-contracted with 30 mM potassium buffer. The curves show dilatation in response to increasing concentrations of αCGRP 10^− 11 to 10^− 7 M in the presence of 66.7 nM of the monoclonal antibody against CGRP (fremanezumab) or an isotype control (KLH). The data show mean ± SEM, n = 2–4 for each concentration. CGRP:calcitonin gene-related peptide; K:potassium; TEV:Teva

Fig. 2

Human intracranial arteries were pre-contracted with 30 mM potassium buffer. The curves show dilatation in response to increasing concentrations of αCGRP 10^− 11 to 10^− 7 M in the presence of 66.7 nM of the monoclonal antibody against CGRP (fremanezumab) or an isotype control (KLH). The data show mean ± SEM, n = 2–4 for each concentration. CGRP:calcitonin gene-related peptide; K:potassium; TEV:Teva

Fig. 3

Human abdominal subcutaneous arteries were pre-contracted with 30 mM potassium buffer. The curves show dilatation in response to increasing concentrations of VIP 10^− 11 to 10^− 7 M in the presence of 66.7 nM of the monoclonal antibody against CGRP (fremanezumab) or an isotype control (KLH). The data show mean ± SEM, n = 2–4 for each concentration. K:potassium; TEV:Teva; VIP:vasoactive intestinal peptide

Fig. 4

Human intracranial arteries were pre-contracted with 30 mM potassium buffer. The curves show dilatation in response to increasing concentrations of VIP 10^− 11 to 10^− 7 M in the presence of 66.7 nM of the monoclonal antibody against CGRP (fremanezumab) or an isotype control (KLH). The data show mean ± SEM, n = 2–4 for each concentration. K:potassium; TEV:Teva; VIP:vasoactive intestinal peptide