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: 9001317
• 负责人: Xuebin Qin
• 金额: $ 32.79万
• 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/9001317
• 关键词: Adjuvant; Alleles; Amino Acids; Antibodies; Antigen Presentation; Antineoplastic Agents; B lymphoid malignancy; Binding; Biological; Blood Circulation; C-terminal; Cancer Patient; Cell Line; Cell surface; Cells; Chronic Lymphocytic Leukemia; Clinical Trials; Colon; Complement; Complement Membrane Attack Complex; Complement-Dependent Cytotoxicity; Computer Simulation; Docking; Epitopes; Erythrocytes; Gastric Adenoma; Half-Life; Health; Hemolysis; Histocompatibility Antigens Class II; Human; Immune response; Immune system; Immunotherapy; In Vitro; Lead; Lymphoma; Major Histocompatibility Complex; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Metabolic; Monoclonal Antibodies; Mutate; N-terminal; Non-Hodgkin' s Lymphoma; Outcomes Research; Parents; Patients; Peptides; Property; Publishing; Relapse; Resistance; Resistance development; Site; Site-Directed Mutagenesis; Solubility; Structure; T-Cell Receptor; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Therapeutic antibodies; Toxic effect; Transgenic Mice; Treatment Efficacy; Up-Regulation; Work; base; cancer cell; cancer therapy; cytotoxicity; genetic regulatory protein; immunogenic; immunogenicity; improved; in vivo; inhibitor/antagonist; malignant breast neoplasm; novel; novel therapeutics; overexpression; pre-clinical; protein aminoacid sequence; 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 regulator 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 to develop a molecule capable of inhibiting hCD59 function in cancer cells and thus facilitate cancer immune 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 have developed a specific and potent hCD59 inhibitor, ILYd4 (114AA), and demonstrated that 1) ILYd4 enhances CDC, thereby sensitizing RTX-resistant NHL cells and primary CLL to RTX or OFA treatment; 2) by itself ILYd4 does not adversely mediate in vivo hemolysis of hCD59-expressing erythrocytes; 3) the sensitivity to CDC effects mediated by RTX or OFA on RTX-resistant NHL cell lines and CLL cells negatively correlated with the level of CD59 on the cell surface; and 4) NHL cells overexpressing hCD59 were directly responsible for the resistance to RTX-mediated CDC. These results rationalize the use of ILYd4 as a lead biological candidate to develop a potential therapeutic adjuvant for RTX treatment of RTX-resistant NHL and CLL. Based on these results, we propose to develop marginally- or non- immunogenic, potent and stable ILYd4-derived peptides. We hypothesize that the optimized ILYd4-derived peptides will improve the physicochemical properties to enable parental administration, eliminate or reduce markedly immunogenicity, increase metabolic stability and half-life in circulation, and enhance therapeutic efficacy, thereby leading to effective ILYd4-related therapeutics that will function as an adjuvant to anti-cancer drugs such as OFA and RTX in B-cell malignancies that have relapsed after prior treatment with Ab-based therapies. Supportively, the availability of crystal structures of hCD59 and ILY enabled us to perform computational docking to identify potential interfaces between ILYd4 and hCD59 and predict nonfunctional and immunogenic epitopes. Recently, we generated a short form of ILYd4 by removing an immunogenic 21 AA residues in N-terminal region (ILYd4del21AA) and found that ILYd4del21AA had an anti-hCD59 activity at the level comparable to parental ILYd4. Therefore, we propose to develop active and deimmunized ILYd4del21AA and pegylate the deimmunized ILYd4del21AA, and to study the immunogenicity, the PK/PD, efficacy and toxicity profile in vivo. Successful outcome of this research will generate a novel therapeutic ILYd4 derivative for clinical trials that will test its efficacy as an adjuvantfor antibody-based cancer therapy such as 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）和奥法木单抗（OFA）在癌症治疗中的功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因子，可限制膜攻击复合物 (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 表达上调是这些癌细胞对 RTX 治疗敏感性的重要决定因素。因此，迫切需要开发一种能够抑制癌细胞中hCD59功能的分子，从而促进癌症免疫治疗。然而，针对 hCD59 的特异性抗体的靶向毒性作用以及 C8 或 C9 肽的较低功效限制了它们的治疗目的。最近，我们开发了一种特异性有效的 hCD59 抑制剂 ILYd4 (114AA)，并证明：1) ILYd4 增强 CDC，从而使 RTX 耐药的 NHL 细胞和原发性 CLL 对 RTX 或 OFA 治疗敏感； 2)ILYd4本身不会不利地介导表达hCD59的红细胞的体内溶血； 3)RTX耐药的NHL细胞系和CLL细胞对RTX或OFA介导的CDC效应的敏感性与细胞表面CD59的水平负相关； 4) 过表达 hCD59 的 NHL 细胞直接导致对 RTX 介导的 CDC 的耐药。这些结果合理化了 ILYd4 作为主要生物候选药物的用途，以开发一种潜在的治疗佐剂，用于 RTX 治疗 RTX 耐药的 NHL 和 CLL。基于这些结果，我们建议开发轻微或无免疫原性、有效且稳定的 ILYd4 衍生肽。我们假设优化的ILYd4衍生肽将改善理化特性，从而能够进行肠胃外给药，消除或显着降低免疫原性，增加代谢稳定性和循环半衰期，并增强治疗效果，从而产生有效的ILYd4相关疗法，作为佐剂发挥作用 抗癌药物（例如 OFA 和 RTX）用于治疗在之前使用基于 Ab 的疗法治疗后复发的 B 细胞恶性肿瘤。 hCD59 和 ILY 晶体结构的可用性使我们能够进行计算对接，以识别 ILYd4 和 hCD59 之间的潜在界面，并预测非功能性和免疫原性表位。最近，我们通过去除N端区域的免疫原性21个AA残基（ILYd4del21AA）生成了ILYd4的短形式，并发现ILYd4del21AA具有与亲本ILYd4相当的抗h​​CD59活性。因此，我们建议开发活性和去免疫的ILYd4del21AA，并将去免疫的ILYd4del21AA聚乙二醇化，并研究其免疫原性、PK/PD、体内功效和毒性特征。这项研究的成功成果将产生 一种用于临床试验的新型治疗性ILYd4衍生物，将测试其作为基于抗体的癌症疗法（例如基于RTX或OFA的B细胞恶性肿瘤疗法）的佐剂的功效，并提供克服毁灭性RTX或OFA耐药性B细胞恶性肿瘤所需的手段。