Manufacture of an intracerebroventricular Enzyme Replacement Therapy for CLN1 Batten Disease

总结 神经元蜡样脂褐质沉积症（NCL）是一组难以治愈的神经退行性疾病 主要影响儿童和年轻人的大脑和视网膜，导致痴呆、失明、癫痫， 和早逝，发病率约为每百万人口1.5至9人（每10万人1.3至7人 出生）。婴儿型CLN 1疾病由CLN 1/PPT 1基因突变引起，该基因编码 溶酶体酶棕榈酰蛋白硫酯酶-1（PPT 1）导致酶减少或缺失 活动CLN 1疾病通常在6至24个月大之间出现，并且有2-3名儿童患有这种疾病。 目前，在美国有24名已知的CLN 1儿童，在巴西有11名（可能有许多人）。 更多未确诊）。目前除了姑息疗法和这种疾病之外，没有其他治疗方法。 是致命的先前已报道在CHO细胞中表达的人重组PPT 1（rhPPT 1）修饰 在PPT 1缺陷小鼠中单次鞘内（IT）和静脉内（IV）给药后的疾病表型 （Ppt1-/-）。在成功完成第一阶段SBIR后，我们达到或超过了IT里程碑， 比较了脑室内（ICV）以及联合给药途径。我们证明了 通过ICV每月一次给予rhPPT 1在Ppt 1-/-小鼠中产生了统计学显著的治疗效果， 作为救援超过60%PPT1酶活性降低二级酶水平，减少损失的 神经元在大脑和脊髓的所有区域，并改善步态和旋转棒的结果。这也说明， 单独通过该途径递送酶可能是足够的，而不是与IT给药组合使用。我们 现在，我已经开发了我们自己的PPT 1细胞系使用CHO-DG 44，这是可扩展的GMP生产， 进行了初步的纯化开发策略，我们正在开发CLN 1的诊断工具。的 CLN 1 ERT数据明确指出，ICV给药是未来研究的理想途径，根据 医生。Collaborations Pharmaceuticals，Inc（CPI）现提议在本II期SBIR中提交preIND (with RTI International和Focal Biosciences的协助），进行PPT 1的制造（使用 Goodwin Biotechnology，Inc.和CMC顾问Stefan Proniuk博士），进行大鼠和犬IND 使毒理学研究（在查尔斯河实验室的协助下），并最终提交IND（与 RTI International和Focal Biosciences的协助）。我们将聘请这些经验丰富的顾问， 临床研究组织在整个过程中协助我们，因为他们有多年的经验。这些 合作将使我们能够成本有效，更快地将这种潜在的治疗方法转化为临床 这可能会挽救患有这种毁灭性疾病的儿童的生命。我们已经获得了一个 rhPPT 1作为生物制品获得FDA的孤儿药认定和罕见儿科疾病认定 对于一种“罕见的儿科疾病”，在FDA批准后， 独家经营7年，并有可能获得罕见儿科疾病券，从而提供回报， 投资（当前价值约1亿美元）。我们与CLN 1家庭接触，经常与他们沟通。 提供最新进展，我们现在已经做好准备继续开发这一潜力 治疗一种毁灭性的疾病。有一个重要的作用，使治疗患者的超罕见的 其他公司，如Biomarin和Ultragenyx，我们在开发rhPPT 1方面的工作 作为CLN 1的ERT，将使我们能够在未来更好地研究罕见疾病并提高CPI。

Summary The neuronal ceroid lipofuscinoses (NCLs) are a group of incurable neurodegenerative storage disorders primarily affecting the brain and the retina of children and young adults, leading to dementia, blindness, epilepsy, and early death, with a prevalence of approximately 1.5 to nine per million population (1.3 to 7 per 100,000 live births). The infantile onset form CLN1 disease is caused by mutations in the CLN1/PPT1 gene, which codes for the lysosomal enzyme palmitoyl-protein thioesterase-1 (PPT1) resulting in a reduction or absence of enzyme activity. CLN1 disease usually presents between 6 and 24 months of age and there are 2-3 children with this form identified each year and currently 24 known children with CLN1 in the US and 11 in Brazil (with likely many more undiagnosed). There are currently no treatments available other than palliative therapies and the disease is fatal. Human recombinant PPT1 (rhPPT1) expressed in CHO cells has been previously reported to modify disease phenotypes following a single intrathecal (IT) and intravenous (IV) administration in PPT1 deficient mice (Ppt1-/-). After successful completion of our Phase I SBIR in which we met or exceeded our milestones IT, intracerebroventricular (ICV) as well as combined routes of delivery were compared. We demonstrated that monthly administration of rhPPT1 via ICV produced statistically significant treatment effects in Ppt1-/- mice, such as rescue of more than 60% PPT1 enzyme activity decreased secondary enzyme levels, decreased the loss of neurons in all regions of brain and spinal cord and improved gait and rotarod results. This also illustrated that delivery of enzyme via this route alone may be sufficient rather than using in combination with IT dosing. We have now developed our own PPT1 cell line using CHO-DG44, that is scalable for GMP manufacture and performed initial purification development strategies and we are developing a diagnostic tool for CLN1. The CLN1 ERT data clearly points to ICV dosing as ideal for future studies, which is also a preferred route according to physicians. Collaborations Pharmaceuticals, Inc (CPI) now proposes in this Phase II SBIR to file a preIND (with the assistance of RTI International and Foresight Biosciences), perform manufacturing of PPT1 (with the assistance of Goodwin Biotechnology, Inc. and CMC consultant Dr. Stefan Proniuk), conduct rat and dog IND enabling toxicology studies (with the assistance of Charles River Laboratories) and ultimately filing an IND (with the assistance of RTI International and Foresight Biosciences). We will hire these experienced consultants and clinical research organizations to assist us throughout the process as they have years of experience. These collaborations will enable us to cost effectively and more rapidly translate this potential treatment to the clinic that can potentially save the lives of children living with this devastating disease. We have already obtained an Orphan Drug Designation and rare pediatric disease designation from the FDA for rhPPT1 as a biological product for a “rare pediatric disease” which offers several benefits in future upon FDA approval, including marketing exclusivity for 7 years and the potential to obtain a rare pediatric disease voucher and thus provide a return on investment (current value ~$100 M). We are engaged with CLN1 families communicating with them frequently to provide progress updates and we are now well positioned to continue the development of this potential treatment for a devastating disease. There is an important role to bring treatments to patients with ultra-rare diseases as illustrated by other companies such as Biomarin and Ultragenyx. Our work on developing rhPPT1 as an ERT for CLN1 would position us well to work on further rare diseases in future and grow CPI.