Novel Targeted Therapy for Breast Cancer

乳腺癌新型靶向治疗

• 批准号: 8004858
• 负责人: Ginette Serrero
• 金额: $ 57.58万
• 依托单位: A AND G PHARMACEUTICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-11 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8004858
• 关键词: Address; Adverse effects; Affect; Antibodies; Antibody Formation; Benign; Binding; Biologic Characteristic; Biological; Breast Cancer Cell; Cell Line; Cell model; Cells; Cellular biology; Chinese Hamster Ovary Cell; Clinical; Clinical Trials; Clone Cells; Cloning; Complementary DNA; Contracts; Development; Disease; Doxorubicin; Drug Formulations; Drug Kinetics; Faslodex(ICI 182,780); Generations; Growth; Growth Factor; Human; Hybridomas; In Vitro; Incidence; Libraries; Light; Malignant Neoplasms; Mammary Gland Parenchyma; Mediating; Metabolic; Mus; Normal tissue morphology; Outcome; PC cell-derived growth factor; Patients; Performance; Phase; Play; Principal Investigator; Production; Productivity; Property; Recombinants; Research; Resistance; Roche brand of trastuzumab; Role; Safety; Serum; Small Business Innovation Research Grant; Tamoxifen; Therapeutic; Tissues; Tumorigenicity; Validation; Woman; advanced disease; angiogenesis; antigen binding; autocrine; base; cancer cell; cancer therapy; cell growth; chimeric antibody; clinical efficacy; design; expression vector; improved; in vivo; innovation; malignant breast neoplasm; malignant phenotype; mortality; neoplastic cell; neutralizing monoclonal antibodies; novel; outcome forecast; overexpression; pre-clinical; programs; public health relevance; research study; scale up; tumor; tumor growth; tumorigenesis

DESCRIPTION (provided by applicant): Breast cancer remains one of the top three cancers to affect and cause mortality in women. Major shortcomings with current treatment are the high level of side effects induced in patients and insufficient efficacy, particularly for patients with metastatic disease. Targeted cancer therapy is designed to treat only the cancer cells and minimize damage to healthy cells. These targets are critical to the tumor's malignant phenotype but not to the host's normal tissues, improving outcomes while minimizing systemic side effects. Anti-HER2, Herceptin, leads the way for targeted therapy in breast cancer with a much lower incidence of side effects and definite efficacy in a specific but small subset of patients. However, there is a clear need for additional therapeutic options including other novel targeted therapies. The 88 kDa autocrine growth factor called PC-Cell Derived Growth Factor (PCDGF), also known as granulin precursor (GP88), has been shown to play a critical role in breast cancer cell biology, exemplified by the following: 1) GP88 is an autocrine growth/survival factor for breast cancer cells, 2) increased GP88 expression in breast cancer cells is associated with increased tumorigenicity, 3) GP88 mediates tumor cell angiogenesis and invasiveness, 4) breast cancer cells overexpressing GP88 are resistant to current therapies - Tamoxifen, Faslodex, doxorubicin, 5) increased GP88 expression in breast cancer tissue correlates with clinical parameters of poor prognosis while normal and benign breast tissue are negative, 6) patients with poor prognosis have elevated GP88 serum levels. These experiments show the essential role that GP88 plays in breast cancer cell tumorigenesis and that the inhibition of GP88 expression or function leads to inhibition of tumor growth. These results highlight the importance of GP88 for targeted therapy of breast cancer via the development of anti-GP88 therapeutic neutralizing monoclonal antibodies (Mabs). Our Phase I research proposed to evaluate a library of murine anti-GP88 for antigen binding properties and for biological activity in GP88-dependent in vitro and in vivo functional tumor cell models. These Phase I specific aims were accomplished and resulted in the identification of anti-GP88 neutralizing Mabs with the in vitro and in vivo potency and efficacy to be candidates for targeted therapy in breast cancer to address a vitally important unmet clinical need. We have selected one Mab, AG1, for further pre-clinical development activities, specifically the generation and characterization of a recombinant Mab (chimerized and humanized Mabs). The following Specific Aims are proposed for this Phase II SBIR application: 1. Cloning of the cDNA encoding the variable sequences of the light chain and heavy chain of the antibody expressed by the AG1 hybridoma cells and generation of expression vectors for both a humanized (HuAG1) and a mouse-human chimeric antibody (ChAG1). 2. Transient expression of the HuAG1 and ChAG1 chimeric antibodies and validation of their binding properties. The antigen binding characteristics and biological activities of the cloned, expressed HuAG1 and ChAG1 will be evaluated and compared with the murine AG1 Mab in order to select candidate recombinant AG1 that fit acceptance criteria for further development. 3. Generate stable HuAG1- or ChAG1-producing cell clones. HuAG1 or ChAG1 selected above will be stably expressed in CHO cells. The HuAG1- or ChAG1-producing clones will be compared for cell growth performance, antibody productivity, and metabolic profile in order to select a candidate clone for scale-up production. 4. Express, purify and characterize the biological properties of the HuAG1 or ChAG1 Mab. The Mab will be evaluated in relevant in vitro and in vivo functional tumor cell models and compared directly with the murine AG1 Mab. GP88 represents a novel, pre-clinically validated target for breast cancer. At the conclusion of this research in this Phase II project, we will have generated a mouse/human chimeric Mab with the in vitro and in vivo pre-clinical efficacy and potency for targeted therapy of breast cancer and we will have developed an optimized clonal antibody production cell line suitable for transfer to a CMO for contract manufacturing. However, once this chimeric anti-GP88 Mab has been generated and characterized it will require further pre-clinical development activities (e.g. formulation, pharmacokinetics and safety) before IND filing for clinical trials in breast cancer. These additional development activities will serve as the basis for a subsequent continuation/renewal Phase II SBIR application. PUBLIC HEALTH RELEVANCE: Breast cancer remains one of the top three cancers to affect and cause mortality in women. Major shortcomings with current treatment are the high level of side effects induced in patients and insufficient efficacy, particularly for patients with advanced disease. The innovative breast cancer therapy to be developed in this proposal will target a mechanism inherent in breast cancer but avoid the side effects associated with many current breast cancer therapies.

描述（由申请人提供）：乳腺癌仍然是影响和导致女性死亡的三大癌症之一。目前治疗的主要缺点是对患者产生高程度的副作用且疗效不足，特别是对于患有转移性疾病的患者。靶向癌症治疗旨在仅治疗癌细胞并尽量减少对健康细胞的损害。这些靶点对于肿瘤的恶性表型至关重要，但对于宿主的正常组织则不然，从而改善结果，同时最大限度地减少全身副作用。抗 HER2 药物赫赛汀引领了乳腺癌靶向治疗的潮流，其副作用发生率低得多，并且对特定但一小部分患者具有明确的疗效。然而，显然需要额外的治疗选择，包括其他新型靶向疗法。 88 kDa 自分泌生长因子称为 PC 细胞衍生生长因子 (PCDGF)，也称为颗粒蛋白前体 (GP88)，已被证明在乳腺癌细胞生物学中发挥着关键作用，例如：1) GP88 是一种乳腺癌细胞的自分泌生长/存活因子，2) 乳腺癌细胞中 GP88 表达增加与致瘤性增加相关，3) GP88 介导肿瘤细胞血管生成和侵袭性，4) 过度表达 GP88 的乳腺癌细胞对当前的治疗方法有耐药性 - 他莫昔芬、Faslodex、阿霉素，5) 乳腺癌组织中 GP88 表达增加与预后不良的临床参数相关，而正常和良性乳腺组织呈阴性，6)预后不良的患者血清GP88水平升高。这些实验表明GP88在乳腺癌细胞肿瘤发生中发挥重要作用，并且抑制GP88表达或功能导致抑制肿瘤生长。这些结果强调了 GP88 通过开发抗 GP88 治疗性中和单克隆抗体 (Mab) 对乳腺癌靶向治疗的重要性。我们的第一阶段研究建议评估鼠抗 GP88 文库的抗原结合特性以及 GP88 依赖性体外和体内功能性肿瘤细胞模型中的生物活性。这些 I 期具体目标已经实现，并导致鉴定出具有体外和体内效力和功效的抗 GP88 中和单克隆抗体，成为乳腺癌靶向治疗的候选药物，以满足至关重要的未满足的临床需求。我们选择了一种单克隆抗体 AG1，用于进一步的临床前开发活动，特别是重组单克隆抗体（嵌合和人源化单克隆抗体）的生成和表征。为此 II 期 SBIR 应用提出以下具体目标： 1. 克隆编码 AG1 杂交瘤细胞表达的抗体轻链和重链可变序列的 cDNA，并生成人源化 (HuAG1) 抗体的表达载体）和鼠人嵌合抗体（ChAG1）。 2. HuAG1 和 ChAG1 嵌合抗体的瞬时表达及其结合特性的验证。将评估克隆、表达的 HuAG1 和 ChAG1 的抗原结合特征和生物活性，并与鼠 AG1 Mab 进行比较，以选择符合进一步开发验收标准的候选重组 AG1。 3. 生成稳定的 HuAG1 或 ChAG1 生产细胞克隆。上述选择的HuAG1或ChAG1将在CHO细胞中稳定表达。将比较产生 HuAG1 或 ChAG1 的克隆的细胞生长性能、抗体生产力和代谢特征，以便选择用于放大生产的候选克隆。 4. 表达、纯化和表征 HuAG1 或 ChAG1 Mab 的生物学特性。 该 Mab 将在相关的体外和体内功能性肿瘤细胞模型中进行评估，并直接与鼠 AG1 Mab 进行比较。 GP88 代表了一种经过临床前验证的新型乳腺癌靶点。在这个二期项目的研究结束时，我们将产生一种小鼠/人嵌合单克隆抗体，具有体外和体内临床前功效和乳腺癌靶向治疗的效力，并且我们将开发一种优化的克隆抗体适合转移至 CMO 进行合同制造的生产细胞系。然而，一旦这种嵌合抗 GP88 Mab 被生成并表征，在乳腺癌临床试验 IND 申请之前，将需要进一步的临床前开发活动（例如配方、药代动力学和安全性）。这些额外的开发活动将作为后续延续/更新第二阶段 SBIR 申请的基础。 公共卫生相关性：乳腺癌仍然是影响和导致女性死亡的三大癌症之一。目前治疗的主要缺点是对患者产生的副作用较高且疗效不足，特别是对于晚期疾病患者。该提案中将开发的创新乳腺癌疗法将针对乳腺癌固有的机制，但避免与许多当前乳腺癌疗法相关的副作用。