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引起女性死亡率的前三名癌症之一。当前治疗的主要缺点是患者引起的高副作用和功效不足，特别是对于转移性疾病的患者。靶向癌症治疗旨在仅治疗癌细胞并最大程度地减少对健康细胞的损害。这些靶标对于肿瘤的恶性表型至关重要，但对宿主的正常组织而言并不重要，从而改善了预后，同时最大程度地减少了全身副作用。抗HER2，Herceptin，为乳腺癌的靶向治疗带来了良好的途径，在特定但较小的患者中，副作用的发生率和明确的功效要低得多。但是，显然需要其他治疗选择，包括其他新型靶向疗法。 88 kDa自分泌生长因子称为PC细胞得出生长因子（PCDGF），也称为颗粒蛋白前体（GP88），已显示在乳腺癌细胞生物学中起着关键作用，以下列为例：1）gp88是GP88是GP88是乳腺癌细胞的乳腺癌细胞，2）与乳腺癌细胞的表达相关，2）介导的肿瘤细胞血管生成和侵袭性，4）过表达GP88的乳腺癌细胞对当前疗法具有抗性-Tamoxifen，Faslodex，faslodex，5）乳腺癌组织中GP88的表达增加了GP88的表达与临床预后较差，而正常乳腺组织的临床参数相关，而良好的乳房组织则不良，而GP的患者却不在5个患者。这些实验表明，GP88在乳腺癌细胞肿瘤发生中起着至关重要的作用，并且抑制GP88表达或功能会导致抑制肿瘤生长。这些结果强调了GP88通过开发抗GP88治疗性中和单克隆抗体（MABS）而对乳腺癌的靶向治疗的重要性。我们的I阶段研究提议评估鼠抗GP88的库中的抗原结合特性，以及在gp88依赖性体外和体内功能性肿瘤细胞模型中的生物学活性。完成了I期特异性目标，并导致鉴定具有体外和体内效力和疗效的抗GP88中和MAB，以应对乳腺癌的有针对性治疗，以满足至关重要的临床需求。我们选择了一个mAB AG1进行进一步的临床前开发活动，特别是重组mAb（嵌合和人源性mAb）的产生和表征。提出了针对此II期SBIR应用的以下具体目的：1。编码由AG1杂交瘤细胞表达的光链和重链的可变序列的cDNA，以及人为化的（HUAG1）和小鼠 - 人类Chimercer Chimeric抗体（Chag1）表达的抗体和表达载体的产生。 2。huag1和chag1嵌合抗体的瞬态表达及其结合特性的验证。将评估克隆，表达的Huag1和Chag1的抗原结合特性和生物学活性，并将其与鼠AG1 MAB进行比较，以选择适合接受接受标准的候选重组AG1，以进行进一步发展。 3。产生稳定的Huag1-或Chag1产生的细胞克隆。上面选择的HUAG1或CHAG1将在CHO细胞中稳定表达。将比较Huag1-或Chag1产生的克隆的细胞生长性能，抗体生产力和代谢谱，以选择候选克隆以进行扩展生产。 4。表达，纯化和表征Huag1或Chag1 mAb的生物学特性。 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代表了一种新型的，经过限制的乳腺癌靶标。在这项II阶段项目的这项研究结束时，我们将生成一个小鼠/人类嵌合MAB，具有体外和体内临床前的临床前疗效和靶向治疗乳腺癌治疗的效力，我们将开发出优化的克隆抗体生产细胞系，适合于转移到CMO进行合同制造的CMO。但是，一旦产生了这种嵌合抗GP88 mAb并表征了表征，它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），然后才能在乳腺癌临床试验中提交IND。这些额外的开发活动将作为随后延续/更新II期SBIR应用的基础。 公共卫生相关性：乳腺癌仍然是影响和引起女性死亡率的前三名癌症之一。当前治疗的主要缺点是患者引起的高副作用和不足的功效，尤其是对于晚期疾病患者。该提案中要开发的创新乳腺癌疗法将针对乳腺癌固有的机制，但避免了与许多当前乳腺癌疗法相关的副作用。