CLFB143 Analogs to Treat Glaucoma

项目摘要 青光眼是世界范围内不可逆性失明的主要原因。在青光眼中，视网膜神经节细胞（RGC）， 将视觉从视网膜传递到大脑的投射神经元受到损伤并死亡， 眼内压（IOP）升高。目前的治疗方法（激光、手术和滴眼液）都是通过降低 IOP。不幸的是，降低IOP可能产生不期望的副作用和/或难以实现。此外，委员会认为， 在一些患者中，尽管IOP显著降低，但RGC损失继续。缺乏“神经保护”剂 直接干扰视网膜神经节细胞的死亡过程。安全有效的开发 神经保护剂将通过补充目前可用的IOP来改善青光眼治疗 治疗方案。为了鉴定这些化合物，约翰霍普金斯的扎克小组开发了一种高- 内容，使用RGC进行高通量小分子筛选。这利用了隔离和 培养原代鼠RGC，然后使用自动荧光成像和生化测定来筛选 通过小分子库，以确定促进RGC存活和神经突的化合物， 结果通过此次筛选，Sutent和VX-680等广谱蛋白激酶抑制剂被 被鉴定为神经保护剂。不幸的是，所鉴定的化合物活性弱且毒性极强。 为了解析出负责细胞死亡和存活的单个激酶， 方法被利用。我们在约翰霍普金斯大学的合作者将他们的主要研究资助局平台改编为高- 通过基于siRNA的筛选，并使用它来筛选靶向小鼠的siRNA阵列文库 kinome。对鉴定的前两个命中（即敲低促进存活的基因）进行了一点探索 激酶DLK及其底物MKK 7。Califia Bio独立设计和合成了类似药物的药物， 该蛋白激酶的抑制剂靶向用于神经病的潜在治疗。强效Califia Bio DLK抑制剂 在约翰霍普金斯的RGC保护试验中非常活跃。我们已经确定了一个潜在的铅系列， 从卡利菲亚生物公司专有支架上筛选出几个匹配结果我们提出了一个药物化学计划， 筛选级联，以将这些筛选结果优化为具有适当体外PK的高级先导化合物 用于直接给药至眼睛以最小化全身毒性风险和有限脱靶活性的曲线。化合物 将被优化以提供足够的药物样分子用于动物中的初步功效测试。同相 第二，我们将优化化合物的眼部给药制剂，药代动力学特性，体内活性， 并在初步毒理学和遗传毒性试验中对其进行表征。

Project Summary Glaucoma is the leading cause of irreversible blindness worldwide. In glaucoma, retinal ganglion cells (RGCs), the projection neurons that transmit vision from the retina to the brain, are injured and die, partially as a result of increased intraocular pressure (IOP). Current therapies (laser, surgery and eye drops) all act by lowering IOP. Unfortunately, lowering IOP can produce undesirable side effects and/or be difficult to achieve. Moreover, in some patients, RGC loss continues despite significant IOP reduction. Lacking are "neuroprotective" agents that directly interfere with the cell death process in RGCs. The development of safe and efficacious neuroprotective agents would improve glaucoma therapy by complementing the currently available IOP treatment options. In order to identify such compounds, the Zack group at Johns Hopkins developed a high- content, high-throughput small molecule screen using RGCs. This takes advantage of the ability to isolate and culture primary murine RGCs and then uses automated fluorescent imaging and biochemical assays to screen through small molecule libraries in order to identify compounds that promote RGC survival and neurite outgrowth. Through this screen broad-spectrum protein kinase inhibitors such as Sutent and VX-680 were identified as neuroprotective. Unfortunately the identified compounds are weakly active and extremely toxic. To parse out the individual kinases responsible for cell death and survival, an RNA interference-based approach was utilized. Our collaborators at Johns Hopkins adapted their primary RGC platform for high- throughput siRNA-based screening and used it to screen an arrayed library of siRNAs targeting the mouse kinome. The top two hits identified (i.e. genes whose knockdown promoted survival) were a little explored kinase, DLK and its substrate, MKK7. Independently, Califia Bio has designed and synthesized drug-like inhibitors of this protein kinase target for potential treatment of neuropathy. Potent Califia Bio DLK inhibitors are very active in the Johns Hopkins' RGC protection assay. We have identified a potential lead series from several screening hits from a Califia Bio proprietary scaffold. We present a medicinal chemistry plan and screening cascade to optimize these screening hits into advanced lead compounds with appropriate in vitro PK profiles for direct dosing to the eye to minimize systemic toxicity risk and limited off-target activity. Compounds will be optimized to provide sufficient drug like molecules for preliminary efficacy tests in animals. In phase two, we will optimize compounds for ocular delivery formulations, pharmacokinetic properties, in vivo activity and characterize them in preliminary toxicology and geneotoxicity assays.