Novel Targeted Therapy for Breast Cancer

乳腺癌新型靶向治疗

• 批准号: 8113461
• 负责人: Ginette Serrero
• 金额: $ 57.81万
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8113461
• 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; ERBB2 gene; 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; 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; humanized monoclonal antibodies; improved; in vivo; innovation; malignant breast neoplasm; malignant phenotype; mortality; neoplastic cell; neutralizing monoclonal antibodies; novel; outcome forecast; overexpression; pre-clinical; 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妇女死亡的三大癌症之一。目前治疗的主要缺点是在患者中引起的高水平副作用和疗效不足，特别是对于转移性疾病患者。靶向癌症治疗的目的是只治疗癌细胞，尽量减少对健康细胞的损害。这些靶点对肿瘤的恶性表型至关重要，但对宿主的正常组织没有作用，从而改善了治疗效果，同时最大限度地减少了全身副作用。抗her2药物赫赛汀（Herceptin）在乳腺癌靶向治疗方面处于领先地位，副作用发生率低得多，对特定但一小部分患者有明确的疗效。然而，显然需要额外的治疗选择，包括其他新的靶向治疗。88 kDa的自分泌生长因子被称为pc细胞衍生生长因子（PCDGF），也被称为颗粒蛋白前体（GP88），已被证明在乳腺癌细胞生物学中发挥关键作用，例如：1) GP88是乳腺癌细胞的自分泌生长/生存因子，2)GP88在乳腺癌细胞中表达增加与致瘤性增加有关，3)GP88介导肿瘤细胞血管生成和侵袭性，4)过表达GP88的乳腺癌细胞对目前的治疗方法-他莫昔芬，法洛德，阿霉素，5) GP88在乳腺癌组织中的表达升高与预后不良的临床参数相关，正常乳腺组织和良性乳腺组织均为阴性；6)预后不良患者血清GP88水平升高。这些实验表明GP88在乳腺癌细胞发生过程中发挥了重要作用，抑制GP88的表达或功能可抑制肿瘤生长。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）在乳腺癌靶向治疗中的重要性。我们的I期研究旨在评估小鼠抗gp88文库在gp88依赖的体外和体内功能肿瘤细胞模型中的抗原结合特性和生物活性。这些I期特定目标已经完成，并导致抗gp88中和单抗的鉴定，具有体外和体内效力和功效，成为乳腺癌靶向治疗的候选药物，以解决一个至关重要的未满足的临床需求。我们已经选择了一种单抗，AG1，用于进一步的临床前开发活动，特别是重组单抗（嵌合和人源化单抗）的生成和表征。第二期SBIR申请的具体目标如下：AG1杂交瘤细胞表达抗体轻链和重链可变序列cDNA的克隆及人源化抗体（HuAG1）和鼠人嵌合抗体（ChAG1）表达载体的生成。2. HuAG1和ChAG1嵌合抗体的瞬时表达及其结合特性的验证。我们将对克隆表达的HuAG1和ChAG1的抗原结合特性和生物学活性进行评价，并与小鼠AG1单抗进行比较，以选择符合接受标准的重组AG1候选蛋白进行进一步开发。3. 生成稳定的产生HuAG1或chag1的细胞克隆。上述选择的HuAG1或ChAG1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、抗体产量和代谢谱，以选择一个候选克隆进行大规模生产。4. 表达、纯化和鉴定HuAG1或ChAG1单抗的生物学特性。该单抗将在相关的体外和体内功能肿瘤细胞模型中进行评估，并直接与小鼠AG1单抗进行比较。GP88代表了一种新的、临床前验证的乳腺癌靶点。在这个II期项目的研究结束时，我们将产生一个具有体外和体内临床前疗效和乳腺癌靶向治疗效力的小鼠/人嵌合单抗，我们将开发一个优化的克隆抗体生产细胞系，适合转移到CMO进行合同制造。然而，一旦这种嵌合抗gp88单抗已经产生并鉴定，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），然后再申请IND用于乳腺癌的临床试验。这些额外的开发活动将作为后续延续/续期第二期SBIR申请的基础。