Vault-CCL21 Nanocapsule for Lung Cancer

描述(申请人提供)：肺癌是美国癌症死亡的主要原因，每年导致的死亡人数超过乳腺癌、前列腺癌、结肠癌、肝癌、肾癌和皮肤癌的总和。从经济角度来看，在美国，2000年因肺癌死亡而损失的生命价值超过2400亿美元，预计2020年将上升到4330亿美元以上。由于肺癌带来的巨大医疗保健和经济负担，改善患者预后的新治疗策略将产生非常重大的影响。切萨皮克Perl公司(C-Perl)和加州大学洛杉矶分校(UCLA)(伦纳德·罗马实验室)的合作者已经鉴定并表征了一种称为拱顶的独特纳米颗粒结构，这种结构高度稳定，在所有高等真核生物中普遍存在。拱顶外壳完全由一种名为主要拱顶蛋白(MVP)的单一蛋白质的多个副本组成。MVP可以很容易地设计成允许附着其他试剂，包括有希望的癌症治疗药物，如趋化因子CCL21。CCL21是一种淋巴趋化因子，对成熟树突状细胞(DC)、幼稚T细胞和记忆性T细胞具有趋化作用。在肺癌动物模型中的临床前研究表明，肿瘤内注射CCL21基因修饰的树突状细胞可以消除肿瘤。Chesapeake Perl，Inc.(C-Perl)已使用PERLXpress蛋白质表达平台将其表达到非常高的水平。这个独特而强大的系统在自动化平台上使用重组杆状病毒在整个昆虫中表达，以经济高效的方式产生高蛋白产量。MVP和CCL21(融合到称为INT的拱顶包装多肽)很容易表达和正确组装，形成CCL21-Vault纳米颗粒。该颗粒已被证明在几天内缓慢释放其趋化因子有效载荷。数百毫克的高纯度CCL21-Vaults可以很容易地制备出来，并在冻干状态下稳定储存，以供未来的疗效评估和测试。在这一第二阶段应用中提出的持续开发将支持高级开发、临床前安全性评估，包括毒理学、有效性和配方，导致IND准备，以支持未来的GCMP产品制造。从杆状病毒感染整个昆虫中获得的重组CCL21-拱顶的肿瘤内给药将在临床前动物模型中进行测试，以销毁肿瘤。CCL21-Vault被提出以避免自体DC制备，最大限度地减少批次之间的变异性，并允许可比性和标准化，以便该颗粒可用作晚期非小细胞肺癌(NSCLC)的现成试剂。

DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer death in the United States and is responsible for more deaths each year than breast, prostate, colon, hepatic, renal and skin cancers combined. Viewed in economic terms, in the United States, the value of life lost from lung cancer deaths in the year 2000 was more than $240 billion, which is estimated to rise to more than $433 billion in 2020. Because of the immense health care and economic burden imposed by lung cancer, new therapy strategies that improve patient outcomes will lead to very a significant impact. Collaborators at Chesapeake PERL, Inc. (C-PERL) and UCLA (Laboratory of Leonard Rome) have identified and characterized a unique nanoparticle structure called a vault that is highly stable, and found ubiquitously in all higher eukaryotes. The vault shell is entirely composed of multiple copies of a single protein called Major Vault Protein (MVP). MVP can be readily engineered to permit attachment of other agents, including promising cancer therapeutics like the chemokine CCL21. CCL21 is a lymphoid chemokine that is chemoattractant for mature dendritic cells (DCs), and naive and memory T cells. Preclinical studies in a lung cancer animal model have demonstrated that intratumoral administration of CCL21 gene-modified dendritic cells led to tumor eradication. Vaults have been expressed at Chesapeake PERL, Inc. (C-PERL) to very high levels using the PERLXpress protein expression platform. The unique and powerful system uses recombinant baculovirus expression in whole insects in an automated platform to generate high protein yields cost effectively. MVP and CCL21 (fused to a vault packaging peptide called INT) are readily expressed and correctly assembled to form CCL21-Vault nanoparticles. The particle has been shown to slowly release its chemokine payload over several days. Hundreds of milligrams of highly purified CCL21-Vaults can be readily prepared and stably stored in a lyophilized state for future therapeutic evaluation and testing. Continued development proposed in this Phase II application will support advanced development, pre- clinical safety evaluation including toxicology, efficacy, and formulation, leading to IND preparation to support future GCMP product manufacture. Intratumoral administration of recombinant CCL21-vaults derived from baculovirus infection of whole insects will be tested in a preclinical animal model for destruction of tumors. The CCL21-Vault is proposed to circumvent autologous DC preparation, minimize batch to batch variability and allow for comparability and standardization so that the particle can be used as an off-the-shelf reagent for advanced non-small cell lung cancer (NSCLC).