Development of Therapeutic CD59 inhibitor for treating B-cell malignancies

开发用于治疗 B 细胞恶性肿瘤的治疗性 CD59 抑制剂

• 批准号: 8725332
• 负责人: Xuebin Qin
• 金额: $ 22.37万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725332
• 关键词: Adjuvant; Antibodies; Antineoplastic Agents; B lymphoid malignancy; B-Lymphocytes; Bacteriophages; Blood Circulation; Body Weight; Cell Line; Cell surface; Cells; Chronic Lymphocytic Leukemia; Clinical Trials; Complement; Complement Membrane Attack Complex; Complement-Dependent Cytotoxicity; Detection; Dose; Erythrocytes; Half-Life; Hemolysis; Human; Immune response; In Vitro; Inhibitory Concentration 50; Lead; Libraries; Lymphoma; Malignant - descriptor; Malignant Neoplasms; Mediating; Metabolic; Mus; Non-Hodgkin' s Lymphoma; Outcomes Research; Parents; Peptides; Phage Display; Property; Publishing; Relapse; Resistance; Series; Solubility; Testing; Therapeutic; Therapeutic antibodies; Toxic effect; Transgenic Mice; Treatment Efficacy; Up-Regulation; Work; base; cancer cell; cancer therapy; cytotoxicity; gene synthesis; genetic regulatory protein; immunogenic; immunogenicity; improved; in vivo; inhibitor/antagonist; novel therapeutics; overexpression; rituximab; therapeutic development

DESCRIPTION (provided by applicant): Rituximab (RTX) and Ofatumumab (OFA) efficacies in cancer therapy depend in part on the induction of complement-dependent cytotoxicity (CDC). Human CD59 (hCD59) is a key complement regulatory protein that restricts the formation of membrane attack complex (MAC), thereby inhibiting induction of CDC. hCD59 is highly expressed in B-cell malignancies such as non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL) and up-regulation of hCD59 expression is an important determinant of the sensitivity of those cancer cells to RTX treatment. Therefore, it is imperative for us to develop a molecule capable of abrogating CD59 function in cancer cells and thus facilitate antibody- mediated cancer therapy. However, targeted toxicity effect from antibody specific against hCD59 and less efficacy of C8 or C9 peptides limit them for therapeutic purposes. Recently, we developed a specific and potent hCD59 inhibitor, rILYd4 (114aa), and demonstrated that rILYd4 enhances CDC in vitro and in vivo, thereby sensitizing RTX resistant lymphoma cells and primary CLL to RTX and OFA treatment. By defining PK/PD profiles of rILYd4 in mice, we showed that by itself rILYd4 does not adversely mediate in vivo hemolysis of hCD59-expressing erythrocytes. We demonstrated that increasing expression level of hCD59 in RTX-resistant cells occurs due to selection of pre-existing clones, rather than de novo induction of hCD59. We also demonstrated that the sensitivity to CDC effects mediated by OFA or RTX on RTX-resistant B-cell malignant cell lines and CLL cells negatively correlated with the level of CD59 on the cell surface. Taken together, our results rationalize the use of rILYd4 as a therapeutic adjuvant for RTX and OFA treatment of RTX-resistant NHL and CLL. Based on these results, we propose to optimize rILYd4 and develop less or non immunogenic, potent and stable rILYd4-derived peptide. We hypothesize that the optimized rILYd4 will improve the physicochemical properties of parental rILYd4, eliminate or reduce markedly immunogenicity, increase metabolic stability and half life in circulation, and enhance therapeutic efficacy, thereb leading to effective rILYd4-related therapeutics for enhancing as a adjuvant to anti-cancer drugs such as OFA and RTX in B-cell malignancies that have relapsed after prior antibody-based therapies. We propose to 1) to identify the sequences in rILYd4 responsible for its anti-hCD59 activity by phage display and synthetic approaches, and use them to develop a small, potent and non-immunogenic rILYd4-derived peptide (derivative) (Aim 1) and 2) to study its immune responses in human HLA transgenic mice and characterize its PK/PD, efficacy and toxicity profiles in humanized hCD59 transgenic mice. (Aim 2). Successful outcome of this research will generate a novel therapeutic rILYd4-derived agent for clinical trials that will test its efficacy a an adjuvant for RTX- or OFA-based B-cell malignancy therapy and provide the much needed means to overcome the devastating RTX- or OFA-resistant B-cell malignancies.

描述(由申请人提供)：利妥昔单抗(RTX)和奥法单抗(OFA)在癌症治疗中的疗效部分取决于补体依赖性细胞毒性(CDC)的诱导。人CD59(HCD59)是一种关键的补体调节蛋白，它限制膜攻击复合体(MAC)的形成，从而抑制CDC的诱导。HCD59在非霍奇金淋巴瘤(NHL)和慢性淋巴细胞白血病(CLL)等B细胞恶性肿瘤中高表达，hCD59表达上调是决定这些肿瘤细胞对RTX治疗敏感性的重要因素。因此，开发一种能够消除癌细胞中CD59功能的分子，从而促进抗体介导的癌症治疗是当务之急。然而，针对hCD59的抗体的靶向性毒性作用以及C8或C9多肽的疗效较差限制了它们的治疗目的。最近，我们开发了一种特异而有效的hCD59抑制剂rILYd4(114aa)，并证明了rILYd4在体内和体外都能增强CDC，从而使RTX耐药淋巴瘤细胞和原发CLL对RTX和OFA治疗敏感。通过在小鼠中定义rILYd4的PK/PD谱，我们表明rILYd4本身并不负面地介导表达hCD59的红细胞在体内的溶血。我们证明，在RTX耐药细胞中，hCD59表达水平的增加是由于选择了先前存在的克隆，而不是从头开始诱导hCD59。我们还证实了对OFA或RTX介导的CDC效应的敏感性与细胞表面CD59的水平呈负相关。综上所述，我们的结果合理地将rILYd4用作RTX和OFA治疗RTX耐药的NHL和CLL的治疗佐剂。基于这些结果，我们建议优化rILYd4，开发免疫原性较低或无免疫原性、有效且稳定的rILYd4衍生肽。我们推测，优化的rILYd4将改善亲本rILYd4的物理化学性质，消除或显著降低免疫原性，增加代谢稳定性和循环半衰期，并提高疗效，从而导致有效的rILYd4相关治疗，以增强作为抗癌药物的辅助，如OFA和RTX，在以前的抗体为基础的治疗后复发的B细胞恶性肿瘤。我们建议1)通过噬菌体展示和合成方法鉴定rILYd4中与其抗hCD59活性有关的序列，并利用这些序列开发一种小的、有效的、非免疫原性的rILYd4衍生多肽(AIM 1)；2)研究其在人HLA转基因小鼠中的免疫应答，并表征其在人源化hCD59转基因小鼠中的PK/PD、疗效和毒性。(目标2)。这项研究的成功结果将产生一种用于临床试验的新型治疗性rILYd4衍生剂，将测试其作为基于RTX或OFA的B细胞恶性肿瘤治疗的佐剂的有效性，并提供克服毁灭性的对RTX或OFA耐药的B细胞恶性肿瘤所需的手段。