Development of a non-Factor small molecule, oral, prophylactic and hemostasis balanced therapy for treatment of clotting disorders including hemophilia A/B

摘要 血友病A、B、C和vwF是遗传性出血性疾病，由以下原因引起： 因子VIII（FVIII）或因子IX（FIX）。基于因子的治疗包括给予 外源性凝血因子浓缩物，目的是达到必要的循环蛋白水平。 然而，基于因子的疗法存在抑制剂（抗体）使治疗剂失活的并发症。 药物，使治疗无效。目前的研究性治疗试图减少抗凝血 天然控制因子蛋白C、抗凝血酶和组织因子蛋白抑制剂（TFPI-抑制剂）， 增强凝血功能受损患者的凝血功能。这种方法具有显著的挑战，因为凝结 这一过程必须保持非常良好的平衡，重要的是要确保预防出血， 增强凝血不会导致意外的血栓形成。此外，这些疗法中的大多数 每两周一次皮下或静脉注射给药，以及与注射剂相关的不良事件 仍然令人担忧。 我们的首要目标是了解传统使用的植物提取物对伤口的显着影响 愈合和止血与血栓形成之间的平衡，以及它们在开发口腔 治疗血友病YewSavin的植物来源的凝结剂和抗凝剂分子对导致 止血和血栓形成的最佳平衡。这种小分子口服，在此类治疗中尚属首例 策略，两种互补成分（CC_2），由查尔酮（促凝血剂）和黄酮（抗凝剂）组成。 凝血剂），这可能会彻底改变血友病和抗血栓治疗凝血和 通过促进血液凝固过程而不引起血栓形成来治疗血栓性疾病。 在初步研究中，凝血和愈合时间的生物测定指导了我们最初的分馏测定， 鉴定促进凝血和愈合的单分子实体。我们对血友病的分子进行了三角测量 使用我们专有的人工智能（AI）、结构匹配（SAR）和 通过生物测定引导的分级能力进行实验验证。使用因子VIII敲除（KO）小鼠 模型，我们证明了我们鉴定的CC_2分子引起凝血时间的显着改善， 在正常大鼠和因子VIII KO小鼠中。这些结果表明，CC_2不依赖于因子VIII， 也比目前使用的一线疗法，即注射因子VIII更有效。最后，目前有 没有已知的抗凝血酶小分子药物。根据我们的初步结果， 似乎是阻断凝血的肝素-抗凝血酶复合物的剂量依赖性逆转。 该项目由YewSavin，Inc.将应用从植物中获得的凝血过程的知识 分子，以解决止血-血栓形成平衡治疗的挑战，并促进发现一种新的抗血栓形成药物。 用于血友病口服预防性治疗的一类小分子、非免疫原性化合物。

ABSTRACT Hemophilia, A, B, C, and vwF are inherited bleeding disorders resulting from a partial or complete deficiency of Factor VIII (FVIII) or Factor IX (FIX), respectively. Factor-based therapies involve the administration of exogenous clotting factor concentrates with the aim of achieving the necessary levels of circulating protein. However, factor-based therapies present the complication of inhibitors (antibodies) inactivating the therapeutic agents, rendering treatments ineffective. Current investigational therapies seek to diminish the anti-clotting natural control factors Protein C, Anti-thrombin and Tissue Factor Protein Inhibitor (TFPI-inhibitor) in order to enhance clotting in patients with impaired clotting. This approach has significant challenges since the coagulation process must remain extremely well-balanced and is important to make sure that prevention of hemorrhage by enhancing clotting does not result in inadvertent thrombosis. Furthermore, most of these therapies are administered in bi-weekly injections, subcutaneous or intravenous, and adverse events relating to injectables remain a concern. Our overarching goal was to understand the remarkable effects of traditionally used plant extracts on wound healing and the balance between hemostasis and thrombosis, and their application in developing an oral treatment for hemophilia. YewSavin’s plant-derived pair of coagulant and anti-coagulant molecules result in the best balance of hemostasis and thrombosis. This small molecule oral, the first in such class, treatment strategy, two Complementary Components (CC_2), consisting of Chalcones (pro-coagulant) and Flavones (anti- coagulant), that could potentially revolutionize hemophilia and anti-thrombotic treatments for clotting and thrombotic disorders alike by promoting blood clotting processes without causing thrombosis. In preliminary studies, bio-assays of clotting and healing times guided our initial fractionation assays towards identifying single molecule entities that promoted clotting and healing. We triangulated molecules for hemophilia treatment and accelerated healing using our proprietary artificial intelligence (AI), structure match (SAR), and experimental verification by bioassay guided fractionation ability. Using a Factor VIII knockout (KO) mouse model, we demonstrated that our identified CC_2 molecules cause significant improvement in clotting time, in normal rats and in Factor VIII KO mice. These results demonstrate that CC_2 is not Factor VIII dependent and is also more efficient than the first line therapy currently used, injection of Factor VIII. Finally, there are currently no known anti-thrombin small molecule agents. According to our preliminary results, the mechanism of action seems to be the dose-dependent reversal of the Heparin-Antithrombin complex that blocks coagulation. This project proposed by YewSavin, Inc. will apply knowledge of clotting processes gained from plant molecules to address the challenges of hemostasis-thrombosis balanced therapy and to advance discovery of a class of small molecule, non-immunogenic, compounds for oral prophylactic treatment in hemophilia.