In the present study, a novel single domain antibody (sdAb) fusion protein, named everestmab, composing of a mutated GLP-1(A8G) fused to the tandem bispecific humanized GLP-1R-targeting and albumin-binding nanobodies was designed and characterized for the therapies for type 2 diabetes mellitus (T2DM). Surface plasmon resonance (SPR) measurements demonstrated everestmab associates with serum albumins of rat and monkey species with high affinity, and tends to be cross-reactive with rat and monkey species. In vitro GLP-1R binding and activation assays revealed that everestmab can specifically activate the GLP-1R, and the antagonist exendin-4 (9-39) did not inhibit the activation yet. In vivo multiple oral glucose tolerance tests (OGTTs) and hypoglycaemic efficacy tests proved that a single injection of everestmab reduced the blood glucose for at least 144 h in Goto-Kakizaki (GK) rats. The plasma half-lives of 4.1 and 7.8 days were observed after a single s.c. administration of everestmab in SD rats and cynomolgus monkeys, respectively. Chronic treatment of everestmab to GK and diet induced obese (DIO) rats achieved beneficial effects on weight reducing, HbA1c lowering, glucose tolerance, liver and pancreas islet function impairment. In summary, everestmab is a unique G-protein-coupled receptor-targeted nanobody fusion protein and exerts potential as a therapeutic treatment for T2DM.

中文翻译：

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Everestmab是一种新型的长效GLP-1 /抗GLP-1R纳米抗体融合蛋白，具有强大的抗糖尿病作用。

在本研究中，设计并表征了一种新颖的单域抗体（sdAb）融合蛋白，名为Everestmab，该蛋白由与串联双特异性人源化GLP-1R靶向和白蛋白结合纳米抗体融合的突变GLP-1（A8G）组成，并针对2型糖尿病（T2DM）的疗法。表面等离子体共振（SPR）测量表明，依维珠单抗与大鼠和猴子物种的血清白蛋白具有高亲和力，并且倾向于与大鼠和猴子物种发生交叉反应。体外GLP-1R结合和激活测定表明，依维珠单抗可以特异性激活GLP-1R，而拮抗剂exendin-4（9-39）尚未抑制该激活。体内多项口服葡萄糖耐量试验（OGTT）和降血糖功效试验证明，单次注射依维珠单抗可在Goto-Kakizaki（GK）大鼠中降低血糖至少144小时。单次皮下注射依维珠单抗分别在SD大鼠和食蟹猴中观察到4.1和7.8天的血浆半衰期。长期使用依维珠单抗治疗GK和饮食诱发的肥胖（DIO）大鼠在减轻体重，降低HbA1c，糖耐量，肝和胰岛功能受损方面取得了有益的效果。总之，everestmab是一种独特的G蛋白偶联受体靶向的纳米抗体融合蛋白，在治疗T2DM方面具有潜力。分别在SD大鼠和食蟹猴中施用Everestmab。长期使用依维珠单抗治疗GK和饮食诱发的肥胖（DIO）大鼠在减轻体重，降低HbA1c，糖耐量，肝和胰岛功能受损方面取得了有益的效果。总之，everestmab是一种独特的G蛋白偶联受体靶向的纳米抗体融合蛋白，在治疗T2DM方面具有潜力。分别在SD大鼠和食蟹猴中施用Everestmab。长期使用依维珠单抗治疗GK和饮食诱发的肥胖（DIO）大鼠在减轻体重，降低HbA1c，糖耐量，肝和胰岛功能受损方面取得了有益的效果。总之，everestmab是一种独特的G蛋白偶联受体靶向的纳米抗体融合蛋白，在治疗T2DM方面具有潜力。