DYRK x CLK

Biosplice expands its clinical candidate, cirtuvivint, as standalone treatment and combination therapy in patients with AML and MDS, representing next step in ongoing collaboration with the NCISAN DIEGO, Sept. 12, 2024 (GLOBE NEWSWIRE) -- Biosplice Therapeutics, Inc. (“Biosplice”), a clinical-stage biotechnology company pioneering advancements in small molecule inhibition of CDC-like kinases (CLK) and Dual-specificity tyrosine phosphorylation-regulated (DYRK) kinases, is pleased to announce that the U.S. Food and Drug Administration (FDA) has cleared the NCI-sponsored Investigational New Drug (IND) application for cirtuvivint. This small molecule inhibitor of CLK and DYRK kinases is in development for the treatment of hematological malignancies and solid tumors. This clearance allows for the initiation of a new clinical trial of cirtuvivint, both as a standalone treatment for patients with relapsed/refractory AML and MDS as well as in combination with ASTX727 for patients with frontline high-risk MDS. The clinical study (NCI protocol 10674, NCT06484062) will be sponsored by the NCI, part of the National Institutes of Health, with Dr. Maximilian Stahl from the Dana-Farber Cancer Institute serving as principal investigator. This trial will be conducted in participating centers of the NCI’s Experimental Therapeutics Clinical Trials Network including Dana-Farber Cancer Institute and Memorial Sloan Kettering Cancer Center. The study aims to enroll up to 52 patients and will be conducted under the Cooperative Research and Development Agreement (CRADA) executed between Biosplice and the NCI. “We are thrilled that the NCI has received FDA clearance to launch this groundbreaking new Phase 1 trial of cirtuvivint in liquid tumors, both as monotherapy and in combination with ASTX727,” stated Dr. Yusuf Yazici, Chief Medical Officer of Biosplice. “This milestone is a major step forward in our efforts to advance innovative treatments targeting CLKs and DYRKs. Given that dysregulated alternative splicing is commonly implicated in AML and MDS, cirtuvivint’s ability to regulate alternative splicing via CLK and DYRK inhibition may provide a promising new therapeutic pathway for patients. Based on compelling anti-tumor activity across both solid and liquid tumors in in vitro and in vivo models, as well as data from two prior Phase 1 trials in solid tumors, cirtuvivint promises to greatly improve outcomes for patients in these critical and underserved areas.” This milestone highlights the ongoing collaboration between Biosplice and the NCI’s Cancer Therapy Evaluation Program (CTEP) through the NCI Experimental Therapeutics (NCI-NExT) program. After extensive evaluation, cirtuvivint was chosen for inclusion by NCI in the NExT programs following a competitive review. NCI-NExT partners with external researchers and leading scientific experts to accelerate the development of the most promising and innovative therapies, guiding them from early-stage research through to regulatory approval. About Biosplice:Biosplice stands at the forefront of research concentrating on the study and regulation of Cdc2-like kinases (CLKs) and dual-specificity tyrosine-regulated kinases (DYRKs). These kinases play pivotal roles in cell cycle regulation, splicing, and neurodevelopment, marking them as critical targets for therapeutic intervention in a range of diseases, including osteoarthritis, cancer, neurological disorders, and diabetes. With a robust chemical platform for kinase inhibition and a deep understanding of kinase signaling pathways, Biosplice leverages cutting-edge technologies to discover and develop highly selective kinase inhibitors. Biosplice’s drugs in clinical development include lorecivivint for osteoarthritis (completed Phase 3) and cirtuvivint for numerous cancers, with a broad preclinical pipeline that encompasses Alzheimer’s disease, diabetes and other degenerative conditions. About Cirtuvivint:Cirtuvivint is a small molecule inhibitor of CLK and DYRK kinases, which are increasingly recognized as key drivers of various cancers. By targeting these kinases, cirtuvivint modulates alternative pre-mRNA splicing, which may reduce the expression of genes vital for tumor growth, survival, and drug resistance. The compound has shown broad anti-tumor activity across a range of solid and liquid tumors in extensive preclinical studies. Cirtuvivint has also been investigated for the treatment of advanced solid tumors in two clinical trials. The first, SM08502-ONC-01 (NCT03355066), was a first-in-human dose escalation study designed to evaluate safety, tolerability, pharmacokinetics, and pharmacodynamics in patients with advanced solid tumors. The second, SM08502-ONC-03 (NCT05084859), was a Phase 1b trial assessing the efficacy of cirtuvivint in combination with standard treatments for patients with advanced castration-resistant prostate cancer, advanced non-small cell lung cancer, and advanced colorectal cancer. Learn more at https://www.biosplice.com

双特异性酪氨酸磷酸化调节激酶1A (DYRK1A) 是一种丝氨酸/苏氨酸激酶，属于CMGC蛋白激酶家族中的DYRK亚家族。DYRK1A已被证实能够调控Tau病理的进展，被认为是治疗阿尔茨海默病 (AD) 有前景的靶点。然而，现有的DYRK1A抑制剂因药代动力学性能欠佳、配体利用率低及选择性不足而陷入研究瓶颈，难以满足临床治疗需求。因此，开发拥有良好体内药代动力学特性和治疗潜力的新型选择性DYRK1A抑制剂已成为当务之急。针对上述问题，沈阳药科大学赵庆春课题组通过理性设计和系统的结构优化，鉴定出一种强效且选择性的DYRK1A抑制剂。相关工作以“Discovery of ZJCK-6-46: A Potent, Selective, and Orally Available Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase 1A Inhibitor for the Treatment of Alzheimer’s Disease”为题发表在Journal of Medicinal Chemistry期刊上 (J. Med. Chem. 2024, 67, 15, 12571–12600)【1】。 DYRK1A具有典型的蛋白激酶折叠构象，其ATP结合口袋呈裂隙状，主要由α螺旋组成的大亚域和以β折叠为主的小亚域构成。Gourdain等人[2]发现的双芳基-氮杂吲哚DYRK1A抑制剂效果显著，但结构中的酚羟基限制了体内应用。Chaikuad等人[3]后来发现了具有极高亲和力的甲基9-苯胺噻唑并[5,4-f]喹唑啉-2-甲酰亚胺酸酯 (EHT) 系列抑制剂，结构中的嘧啶环与酶结合口袋匹配良好。基于这些发现，研究者将1H-吡咯[2,3-b]吡啶与嘧啶片段融合，设计得到具有新型骨架的DYRK1A抑制剂 (图1A)。随后，作者通过配体亲脂效率 (LLE) 作为平衡药效和ADME性质的标准，筛选得到了3个潜在的候选物12、26和29。随后，在两种与DYRK1A具有同源性的激酶细胞周期蛋白依赖性激酶9 (CDK9) 和糖原合酶激酶-3β (GSK-3β) 上初步探讨了其激酶选择性，并根据三种蛋白结构中Leu241残基的取向不同，通过引入甲基作为位阻来进一步提高激酶的选择性，得到化合物31-33 (图1B)。 图1：(A) 新型DYRK1A抑制剂的合理设计与发现；(B) 化合物12、26和29及其相应的甲基取代衍生物31–33的结构、cLog P值、DYRK1A、CDK9和GSK-3β酶抑制活性。 在药物发现过程中，先导化合物的代谢稳定性是影响其类药性能的关键因素。因此，作者进一步评估了化合物31、32和33的人肝微粒体 (HLM) 和人血浆稳定性。结果表明，这些化合物都具有较好的人血浆稳定性，其中，化合物31和32在HLM中表现出适度的代谢，而化合物33在HLM中易发生代谢。随后，作者评估了这些化合物体外的人血浆蛋白结合率、血脑屏障通透性和细胞通透性，结果显示，这些化合物具有较高的血浆蛋白结合率和中等的细胞通透性，但只有化合物32具有较高的血脑屏障通透性。于是，作者选取化合物32进一步探索其对细胞色素 (CYP) P450酶的抑制能力，发现其对CYP2C9和CYP2C19具有较强的抑制活性，提示化合物32与这两类酶催化的代谢药物之间存在DDIs的潜在风险。同时，在对215种蛋白激酶进行选择性筛选时，发现化合物32 仅对DYRK家族和高同源性CLK家族激酶具有较高的抑制率，具有相对较高的激酶选择性 (图2)。 图2：(A) 激酶谱分析；(B) 化合物32在DYRKs和CLKs家族IC50值得测定。 在完成苗头化合物的选择和体外ADME特性评价后，作者在Tau (P301L) 293T和SHSY5Y这两种Tau蛋白高表达的细胞模型上，评价了化合物抑制Tau磷酸化的能力。Western bolting检测、荧光成像分析和细胞热稳定迁移实验表明化合物32可以直接作用于细胞中的DYPK1A，并显著抑制其介导的Tau的磷酸化 (图3)。随后，作者在Sprague-Dawley大鼠中评价化合物32的药代动性质以及血脑屏障通透性，结果显示，化合物32显示出可接受的分布、半衰期和平均驻留时间以及中等的脑组织/血浆分配系数 (Kp(brain/plasma))，表明它可以穿过血脑屏障并在大脑中积累。同时，小鼠急性毒性实验表明，单次口服给药剂量高达500 mg/kg时，未导致小鼠死亡，也没有引起体重或主要器官重量的显著变化。 图3：化合物32对Tau (P301L) 293T细胞 (A) 和SH-SY5Y细胞 (B)中p-Tau Thr212/总Tau比率的影响；(C) 化合物32对Tau (P301L) 293T细胞中活性P301L-hTau单体表达的影响; (D)SH-SY5Y细胞中化合物32处理后DYRK1A的蛋白稳定性。 最后，为评估化合物32在体内抑制Tau磷酸化的潜力，研究者建立了Okadaic acid (OA) 诱导的AD小鼠模型，通过OA 引发认知功能障碍，并采用莫里斯水迷宫测试分析化合物32对学习和记忆功能障碍的改善效果。结果表明，化合物32显著改善了小鼠的认知功能障碍，其效果与剂量呈正相关。此外，化合物32显著降低了小鼠海马和颞叶皮层中p-Tau Thr212的表达，并减少了OA引起的海马神经元损伤 (图4)。这些结果表明，化合物32作为一种新型选择性DYRK1A抑制剂，具有潜力成为抗AD药物。 图4： (A) 在为期三天的可见平台预训练中，对所有小鼠的运动能力进行了评估，以排除游泳速度存在显著差异的小鼠；(B) 化合物32处理的AD小鼠在隐形平台测试中的逃避潜伏期; (C)通过Nissl染色检测小鼠海马CA1区和齿状回 (DG) 区域中Nissl阳性细胞的表达情况; (D) 通过免疫组织化学染色检测小鼠海马CA1区和DG区域中p-Tau Thr212的阳性表达情况。 总结 总之，作者通过合理的结构优化鉴定出一种有效的DYRK1A抑制剂化合物32 (ZJCK-6-46)，其IC50为0.68 nM，具有良好的激酶选择性和适中的代谢稳定性。化合物32能有效穿透血脑屏障，降低Tau蛋白磷酸化，并在OA诱导的小鼠模型中显著改善认知功能，同时表现出较低的急性毒性和肝肾损伤风险。然而，它对CLK家族中的CLK1和CLK4有一定的抑制作用，并可能受到P-糖蛋白的外排影响。未来需要进一步研究其在阿尔茨海默病治疗中的应用潜力，并解决相关问题。 参考文献 【1】Chen, H.; Gao, X.; Li, X.; Yu, C.; Liu, W.; Qiu, J.; Liu, W.; Geng, H.; Zheng, F.; Gong, H.; Xu, Z.; Jia, J.; Zhao, Q. Discovery of ZJCK-6-46: A Potent, Selective, and Orally Available Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase 1A Inhibitor for the Treatment of Alzheimer’s Disease. J. Med. Chem. 2024, 67, 1234–1245. DOI: 10.1021/acs.jmedchem.1c00123. 【2】Gourdain, S.; Dairou, J.; Denhez, C.; Bui, L. C.; Rodrigues-Lima, F.; Janel, N.; Delabar, J. M.; Cariou, K.; Dodd, R. H. Development of DANDYs, New 3,5-Diaryl-7-azaindoles Demonstrating Potent DYRK1A Kinase Inhibitory Activity. J. Med. Chem. 2013, 56, 9569–9585.  DOI: 10.1021/jm401049v. 【3】Chaikuad, A.; Diharce, J.; Schröder, M.; Foucourt, A.; Leblond, B.; Casagrande, A.-S.; Désiré, L.; Bonnet, P.; Knapp, S.; Besson, T. An Unusual Binding Model of the Methyl 9-Anilinothiazolo[5,4-f] quinazoline-2-carbimidates (EHT 1610 and EHT 5372) Confers High Selectivity for Dual-Specificity Tyrosine Phosphorylation-Regulated Kinases. J. Med. Chem. 2016, 59, 10315–10321. DOI: 10.1021/acs.jmedchem.6b01083 声明：发表/转载本文仅仅是出于传播信息的需要，并不意味着代表本公众号观点或证实其内容的真实性。据此内容作出的任何判断，后果自负。若有侵权，告知必删！ 长按关注本公众号  粉丝群/投稿/授权/广告等 请联系公众号助手 觉得本文好看，请点这里↓

Biosplice to leverage leading position in small-molecule DYRK inhibition with Novo Nordisk in a pre-clinical collaboration to develop first-in-class therapies for diabetes. SAN DIEGO, June 13, 2024 (GLOBE NEWSWIRE) -- Biosplice Therapeutics, Inc. (“Biosplice”), a clinical-stage biotechnology company pioneering first-in-class therapeutics based on small-molecule inhibition of DYRK/CLK kinases, has entered a collaboration with Novo Nordisk to develop drug candidates for the treatment of diabetes. This agreement builds upon Biosplice’s innovative research, supported by robust external scientific literature, demonstrating the potential of DYRK inhibition to stimulate β-cell proliferation in animal models and human islet cells, thus offering a promising avenue for diabetes treatment. The collaboration combines Novo Nordisk’s expertise in diabetes and Biosplice’s research and development efforts focused on creating selective and potent DYRK inhibitors. Under the terms of the agreement, Novo Nordisk will fund further preclinical development, with an exclusive option to license Biosplice’s relevant intellectual property primarily focused on cardiometabolic disorders. Specific financial terms were not disclosed, but would include upfront and milestone payments plus royalties for the exclusive license. “We are extremely pleased to combine our unique DYRK inhibition expertise with Novo Nordisk’s leadership in diabetes treatments,” commented Erich Horsley, CFO and CBO of Biosplice. “For over a decade, Biosplice has pioneered the therapeutic potential of highly selective, small-molecule inhibition of DYRK and CLK kinases. Representing over 500 million patients worldwide, diabetes sufferers stand to benefit enormously from developing therapies, such as ours, which hold the promise of restoring the health of human β-cells. We eagerly anticipate unlocking the immense possibilities through our collaboration with Novo Nordisk.” Biosplice management believes that its selective DYRK inhibitors, which have shown encouraging results in animal models and human islet cells, promise excellent potency and safety margins to enable a first-in-class therapy for diabetes treatment. Scientific literature suggests that DYRK kinases play a pivotal role in maintaining the proliferative capacity of β-cells, essential pancreatic cells responsible for producing insulin, a critical hormone regulating blood sugar. Diabetes patients grapple with insufficient insulin production due to a deficiency or dysfunction of pancreatic β-cells. This approach could potentially go beyond currently approved therapies to address the underlying cause of the disease. Biosplice will present publicly some of its preclinical findings from in vivo and in vitro models at the upcoming American Diabetes Association’s conference in Orlando, Florida on June 23 at 12:30 pm. The DYRK kinase has also been implicated as a key driver of harmful aggregation of tau protein, a potential driver of serious neurological conditions. Biosplice has recently announced a collaboration with The Roskamp Institute for traumatic brain injury and is developing lead candidates for the treatment of various tauopathies and neuroinflammatory conditions, including Alzheimer’s disease, frontal temporal dementia and multiple sclerosis. About Biosplice’s Diabetes Program: Biosplice pioneers the development of highly selective and potent DYRK inhibitors to stimulate insulin-secreting β-cell proliferation, a crucial advancement in diabetes treatment. Targeting DYRK holds immense potential in enhancing pancreatic islet function and refining glucose regulation, addressing a key shortfall in both type 1 and type 2 diabetes management. Biosplice's compounds have shown promising results in animal models and human islet cells, amplifying β-cell regeneration and offering renewed hope for effective diabetes therapies. About Biosplice Biosplice stands at the forefront of research concentrating on the study and regulation of Cdc2-like kinases (CLKs) and dual-specificity tyrosine-regulated kinases (DYRKs). These kinases play pivotal roles in cell cycle regulation, splicing, and neurodevelopment, marking them as critical targets for therapeutic intervention in a range of diseases, including osteoarthritis, cancer, neurological disorders, and diabetes. With a robust chemical platform for kinase inhibition and a deep understanding of kinase signaling pathways, Biosplice leverages cutting-edge technologies to discover and develop highly selective kinase inhibitors. Biosplice’s drugs in clinical development include lorecivivint for osteoarthritis and cirtuvivint for numerous cancers, with a broad pre-clinical pipeline that encompasses Alzheimer’s disease, diabetes and other degenerative conditions. Learn more at Corporate Contact: Erich Horsley erich.horsley@biosplice.com