Humanized monoclonal antibodies to treat mucormycosis

人源化单克隆抗体治疗毛霉菌病

• 批准号: 9759762
• 负责人: ASHRAF S. IBRAHIM
• 金额: $ 29.98万
• 依托单位: VITALEX BIOSCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-09 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759762
• 关键词: Adrenal Cortex Hormones; Affinity; Allergic Reaction; Amino Acids; Angioinvasion; Animal Model; Antibiotics; Antibodies; Antibody Therapy; Antifungal Agents; Antifungal Therapy; Antigens; Attenuated; Binding; Biological Sciences; Blood Vessels; Brain; Cell Line; Cell Surface Proteins; Clinical; Clinical Trials; Cunninghamella; Data; Development; Diabetes Mellitus; Diabetic Ketoacidosis; Diagnostic; Disease; Disease Progression; Dose; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Excision; Fc Immunoglobulins; Feasibility Studies; Funding; Genes; Goals; Grant; Heat shock proteins; Heat-Shock Response; Hematogenous; Hematopoietic Stem Cell Transplantation; Human; Human Cloning; IgG1; Immune system; Immunocompromised Host; Immunotherapy; In Vitro; Incidence; Infection; Inflammation; Injury; Intervention; Invaded; Investments; Iron; Laboratories; Lead; Life; Mammalian Cell; Mediating; Molds; Monoclonal Antibodies; Mucor; Mucorales; Mucormycosis; Mus; Mycoses; Necrosis; Neutropenia; Operative Surgical Procedures; Organ Transplantation; Organism; Orphan Drugs; Outcome; Pathogenesis; Pathway interactions; Patients; Penetration; Peptides; Phagocytosis; Pharmaceutical Preparations; Phase; Positioning Attribute; Prevalence; Prevention strategy; Privatization; Process; Proteins; Reporting; Research; Resources; Rhizomucor; Rhizopus; Risk; Risk Factors; Serum; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Soldier; Stains; Technology; Testing; Therapeutics for Rare and Neglected Diseases; Thrombosis; Tissues; Toxic effect; Trauma; Umbilical vein; United States National Institutes of Health; Vaccines; Virulence; Work; Zygomycosis; base; cGMP production; cancer transplantation; cell injury; cell type; clinical development; clinically relevant; combat; cross reactivity; driving force; effective therapy; glucose-regulated proteins; human tissue; humanized monoclonal antibodies; immunoreaction; improved; in vitro activity; in vivo; injured; lead candidate; method development; mortality; mortality risk; mouse model; murine monoclonal antibody; mutant; novel; novel therapeutics; operation; pharmacokinetics and pharmacodynamics; polyclonal antibody; posaconazole; pre-clinical; prevent; primary endpoint; programs; secondary endpoint; synergism; treatment strategy

Mucormycosis, most commonly caused by Rhizopus oryzae, is a life-threatening infection that occurs in patients immunocompromised by diabetic ketoacidosis (DKA), neutropenia, corticosteroid use, increased serum iron and/or severe trauma. Because of the rising prevalence of these risk factors, the incidence of mucormycosis has risen. Despite disfiguring surgery and aggressive antifungal therapy, the mortality of mucormycosis ranges from ~ 50% to 100%. The obvious unmet need for new, effective treatments and preventive strategies has been the driving force of our research program for over seventeen years. We propose to develop a passive vaccine targeting mucormycosis (i.e., an antibody that can be administered to patients with mucormycosis). Data in the academic laboratory of our founder (Dr. Ibrahim) indicate that antibody-based therapy is a promising strategy to treat mucormycosis. This technology is based on the important discovery that the fungal cell surface proteins encoded by CotH facilitate disease progression by allowing R. oryzae to invade mammalian cells via binding to Glucose Regulated Protein 78 (GRP78), a heat shock conserved protein expressed on endothelial cells lining blood vessels during mucormycosis. Importantly, CotH proteins were found to be conserved among Mucorales (organisms that cause mucormycosis) and absent from any other cell type including mammalian. Our data also show that CotH proteins are key determinants of mucormycosis pathogenesis since R. oryzae coth null mutants have markedly reduced virulence in mouse models of mucormycosis. Further, polyclonal antibodies targeting CotH are highly protective against murine mucormycosis caused by R. oryzae, Mucor, Lichtheimia, Cunninghamella, Rhizomucor, and Apophysomyces. Importantly and highly relevant to this application, anti-CotH murine monoclonal antibodies (mAb) raised against a peptide predicted to be present in the binding domain to GRP78, prevent the ability of R. oryzae to invade and injure endothelial cells in vitro and protect mice from mucormycosis caused by several Mucorales to levels that exceed those seen with antifungal therapy. While these mAbs are a promising new therapy for mucormycosis, the feasibility of further clinical development will hinge upon successful humanization of the Abs. Mouse mAbs cannot be used to treat humans, because humans mount an immune reaction to mouse mAbs that can cause rapid removal of the mAbs, systemic inflammation, severe allergic reactions, and even a risk for death. The humanization process prevents these undesirable effects. Thus, we propose two AIMS: 1) Develop our lead murine mAb into a humanized version with retained/enhanced binding ability to CotH proteins; and 2) Determine the protective activity of the humanized Ab in vitro/in vivo and evaluate its toxicity to human tissues. We propose conservative feasibility milestones that are part of a standard, methodical development pathway for our unique mAbs as a novel treatment for mucormycosis. The proposed work will identify a lead humanized Ab that will go into further development to ultimately test in clinical trials as an adjunctive therapy.

毛霉菌病，最常见的是由根霉引起，是一种危及生命的感染，发生在病人 糖尿病酮症酸中毒（DKA）、中性粒细胞减少症、皮质类固醇使用、血清铁升高导致免疫功能低下 和/或严重创伤由于这些危险因素的流行率不断上升，毛霉菌病的发病率已 上升。尽管进行了毁容手术和积极的抗真菌治疗，毛霉菌病的死亡率从 ~ 50%至100%。对新的、有效的治疗和预防策略的明显未满足的需求是 我们的研究计划的驱动力超过十七年。 我们建议开发一种针对毛霉菌病的被动疫苗（即，一种抗体， 给予患有毛霉菌病的患者）。数据在我们的创始人的学术实验室（博士.易卜拉欣） 表明基于抗体治疗是治疗毛霉菌病的有前景的策略。该技术基于 CotH编码的真菌细胞表面蛋白通过以下方式促进疾病进展的重要发现： 允许R.通过结合葡萄糖调节蛋白78（GRP 78），一种热蛋白， 毛霉菌感染时血管内皮细胞表达休克保守蛋白重要的是， 发现CotH蛋白在毛霉目（引起毛霉病的生物体）中是保守的，并且不存在CotH蛋白。 包括哺乳动物在内的任何其他细胞类型。我们的数据还表明，CotH蛋白质是 毛霉菌病发病机制，因为R.无突变体在小鼠中的毒力显著降低 毛霉菌病的模型。此外，靶向CotH的多克隆抗体对鼠免疫具有高度保护性。 毛霉菌病由R.真菌属、毛霉属、Lichtheimia、小克银汉霉属、根毛霉属和顶囊霉属。 与本申请重要且高度相关的是，抗CotH鼠单克隆抗体（mAb）针对 预测存在于GRP 78的结合结构域中的肽阻止R.入侵和 体外损伤内皮细胞并保护小鼠免受由几种毛霉菌引起毛霉菌病的影响， 超过了抗真菌治疗。虽然这些单克隆抗体是毛霉菌病的一种有前途的新疗法， 进一步临床开发的可行性将取决于Ab的成功人源化。小鼠mab 不能用于治疗人类，因为人类对小鼠mAb产生免疫反应， mAb的快速去除、全身炎症、严重过敏反应，甚至死亡风险。的 人源化过程防止这些不期望的作用。因此，我们提出两个目标：1）开发我们的领先优势 将鼠mAb转化为具有保留的/增强的与CotH蛋白的结合能力的人源化形式;和2）确定 人源化抗体的体内外保护活性，并评价其对人体组织的毒性。 我们提出保守的可行性里程碑，这是一个标准的一部分，有条不紊的发展 我们独特的单克隆抗体作为毛霉菌病的新治疗途径。拟议的工作将确定一个牵头机构， 人源化Ab将进入进一步开发，最终在临床试验中作为免疫疗法进行测试。