Kopruszinski Caroline M, Linley John E, Thornton Peter, Walker Alison S, Newton Philip, Podichetty Sadhana, Ruparel Radhey Hemendra, Moreira de Souza Luiz Henrique, Navratilova Edita, Meno-Tetang Guy, Gurrell Ian, Dodick David W, Dobson Claire, Chessell Tharani, Porreca Frank, Chessell Iain
Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USA.
Neuroscience, BioPharmaceutical R&D, AstraZeneca, Cambridge, CB2 0AA, UK.
Brain. 2025 Apr 3;148(4):1345-1359. doi: 10.1093/brain/awae344.
Protease activated receptor 2 (PAR2) is a G-protein coupled receptor expressed in meningeal neurons, fibroblasts and mast cells that may be targeted to treat migraine. MEDI0618, a fully humanized PAR2 monoclonal antibody, engineered to enhance FcRn-dependent recycling and currently in clinical development, was evaluated in human and rodent in vitro assays, in multiple murine in vivo migraine models and in a model of post-traumatic headache. MEDI0618 bound specifically and with high affinity to cells expressing human PAR2 (hPAR2) and prevented matriptase-induced increase in cytosolic calcium. Similarly, MEDI0618 prevented matriptase-induced calcium in primary fibroblasts and microvascular endothelial cells from human dura mater. MEDI0618 had no effect on hPAR1 receptors. Single-cell calcium imaging of acutely dissociated mouse trigeminal ganglion neurons confirmed expression and functionality of mouse PAR2. Studies in vivo used evoked cutaneous allodynia as a surrogate of headache-like pain and, in some experiments, rearing as a measure of non-evoked headache pain. MEDI0618 was administered subcutaneously to C57BL6/J female mice prior to induction of migraine-like pain with (i) systemic nitroglycerin or compound 48/80 (mast cell degranulator); or (ii) with supradural compound 48/80 or an inflammatory mediator (IM) cocktail. To assess possible efficacy against CGRP receptor (CGRP-R)-independent pain, MEDI0618 was also evaluated in the IM model in animals pretreated with systemic olcegepant (CGRP-R antagonist). Migraine-like pain was also induced by inhalational umbellulone, a TRPA1 agonist, in animals primed with restraint stress in the presence or absence of MEDI0618 as well as in a model of post-traumatic headache pain induced by a mild traumatic brain injury. MEDI0618 prevented cutaneous allodynia elicited by systemic nitroglycerin, compound 48/80 and from supradural compound 48/80 and IM. Systemic olcegepant completely blocked periorbital cutaneous allodynia induced by supradural CGRP but failed to reduce IM-induced cutaneous allodynia. In contrast, MEDI0618 fully prevented IM-induced cutaneous allodynia, regardless of pretreatment with olcegepant. Umbellulone elicited cutaneous allodynia only in restraint stress-primed animals, which was prevented by MEDI0618. MEDI0618 prevented the decrease in rearing behaviour elicited by compound 48/80. However, MEDI0618 did not prevent mild traumatic brain injury-related post-traumatic headache measures. These data indicate that MEDI0618 is a potent and selective inhibitor of PAR2 that is effective in human and rodent in vitro cell systems. Further, blockade of PAR2 with MEDI0618 was effective in all preclinical migraine models studied but not in a model of post-traumatic headache. MEDI0618 may represent a novel therapy for migraine prevention with activity against CGRP-dependent and independent attacks.
蛋白酶激活受体2(PAR2)是一种G蛋白偶联受体,在脑膜神经元、成纤维细胞和肥大细胞中表达,可能是治疗偏头痛的靶点。MEDI0618是一种完全人源化的PAR2单克隆抗体,经设计可增强FcRn依赖性再循环,目前正处于临床开发阶段,已在人和啮齿动物的体外试验、多个小鼠体内偏头痛模型以及创伤后头痛模型中进行了评估。MEDI0618与人PAR2(hPAR2)表达细胞特异性结合且亲和力高,并能阻止matriptase诱导的细胞内钙增加。同样,MEDI0618可阻止matriptase诱导人硬脑膜原代成纤维细胞和微血管内皮细胞内的钙增加。MEDI0618对hPAR1受体无影响。急性分离的小鼠三叉神经节神经元的单细胞钙成像证实了小鼠PAR2的表达和功能。体内研究使用诱发的皮肤异常性疼痛作为类头痛疼痛的替代指标,在一些实验中,使用竖毛反应作为非诱发头痛疼痛的指标。在用(i)全身给予硝酸甘油或化合物48/80(肥大细胞脱颗粒剂);或(ii)硬膜上给予化合物48/80或炎症介质(IM)混合物诱导类偏头痛疼痛之前,将MEDI0618皮下注射给C57BL6/J雌性小鼠。为了评估对降钙素基因相关肽受体(CGRP-R)非依赖性疼痛的可能疗效,还在全身给予olcegepant(CGRP-R拮抗剂)预处理的动物的IM模型中评估了MEDI0618。在有或没有MEDI0618的情况下,吸入TRPA1激动剂伞形酮也在经束缚应激预处理的动物中诱发类偏头痛疼痛,以及在轻度创伤性脑损伤诱导的创伤后头痛疼痛模型中诱发类偏头痛疼痛。MEDI0618可预防全身硝酸甘油、化合物48/80以及硬膜上化合物48/80和IM引起的皮肤异常性疼痛。全身给予olcegepant可完全阻断硬膜上给予CGRP诱导的眶周皮肤异常性疼痛,但未能减轻IM诱导的皮肤异常性疼痛。相比之下,无论是否用olcegepant预处理,MEDI0618均可完全预防IM诱导的皮肤异常性疼痛。伞形酮仅在经束缚应激预处理的动物中诱发皮肤异常性疼痛,MEDI0618可预防这种疼痛。MEDI0618可预防化合物48/80引起的竖毛行为减少。然而,MEDI0618不能预防轻度创伤性脑损伤相关的创伤后头痛指标。这些数据表明,MEDI0618是一种有效的PAR2选择性抑制剂,在人和啮齿动物的体外细胞系统中均有效。此外,用MEDI0618阻断PAR2在所有研究的临床前偏头痛模型中均有效,但在创伤后头痛模型中无效。MEDI0618可能代表一种预防偏头痛的新疗法,对CGRP依赖性和非依赖性发作均有活性。