Notch-4

Eli Lilly currently holds the throne as the world's most valuable pharmaceutical company by market capitalization. On July 15, 2024, Eli Lilly's stock price reached a historical high of $966.1 per share, with a company valuation of $910 billion, getting closer to becoming the first trillion-dollar valued pharmaceutical company. Eli Lilly's financial report for 2023 shows that its revenue mainly comes from dulaglutide ($7.133 billion), tirzepatide ($5.163 billion), abemaciclib ($3.863 billion), and ixekizumab ($2.76 billion), among others. With the continued volume growth of tirzepatide and the market launch of the new Alzheimer's drug Donanemab, the future remains promising. Eli Lilly's success is closely related to its inherent pragmatism. Although many of its products are not the first to be approved and launched, Eli Lilly's drugs are always present in popular targets and competitive fields, such as CDK4/6, IL-17, IL-23, and GLP-1. Eli Lilly's acquisitions are also discerning; for example, in September 2018, it acquired the GLP1 small molecule agonist Orforglipron from the Japanese company Chugai, which was still in the preclinical stage at the time. As Pfizer announced the discontinuation of the development of lotiglipron due to liver toxicity issues, Eli Lilly's Orforglipron has taken the lead globally, advancing to Phase 3 first. Compared to Eli Lilly's various successful acquisition cases, Gilead's acquisitions outside the viral field have almost entirely failed. For R&D personnel, this may serve as a guide to avoiding pitfalls. In the synapse database's transaction section, by selecting Eli Lilly as the transferee, one can see 129 transaction records. Among them, the transaction with PRISM BioLab on November 28, 2023, caught the author's attention. This licensing and cooperation agreement, which may total up to $660 million, aims to jointly develop oral small molecule drugs. PRISM BioLab utilizes its proprietary PepMetics technology to discover orally available small molecule inhibitors targeting protein-protein interaction (PPI) pathways, to treat patients with cancer, autoimmune, fibrotic, and other diseases. PepMetics are a unique class of small molecules that mimic the three-dimensional structures of α-helices and β-turns, which are common peptide structures in cellular PPIs and receptor-ligand interactions. According to the agreement, the two companies will collaborate to discover small molecule inhibitors of PPI targets selected by Eli Lilly through PRISM's proprietary PepMetics technology. Eli Lilly has the option to add two more targets to the collaboration and is responsible for the clinical development and commercialization of the resulting products. Although PRISM has already established cooperation with large pharmaceutical companies such as Eisai, Boehringer Ingelheim, Merck KGaA, and Roche, no intelligence information related to MNC companies has been found in either the pipeline or patent applications. In the patent section of the Synapse database, searching for patents applied for by PRISM BioLab yields over 211 entries. By limiting the publication date to the most recent five-year period, one can get a general sense of the company's research and development activities. The first six patents published include WO2023200017, WO2023140369, WO2023090366, WO2022158610, WO2022075486, and WO2022014724; half of these patents are related to inhibitors of the Notch signaling pathway. The Notch signaling pathway is an evolutionarily conserved mechanism that plays an indispensable role in the development and tissue homeostasis of mammals. Notch receptors and ligands contain single-pass transmembrane domains and are expressed on the cell surface; therefore, Notch signaling is particularly important in mediating communication between adjacent cells that express the receptors and ligands. In rodents and humans, four known Notch receptors have been identified, termed Notch 1 to Notch 4. Notch receptors are heterodimeric proteins composed of extracellular and intracellular domains that are initially synthesized as a single polypeptide. The receptor-ligand interaction triggers a series of proteolytic cleavages of the Notch receptor polypeptide, involving the activity of γ-secretase. The activity of γ-secretase cleaves the Notch intracellular domain (NICD) on the inner side of the plasma membrane, which then translocates to the nucleus to form a transcription factor complex. The Notch intracellular domain (NICD) is the active form of the protein. Various functions of Notch signaling include proliferation, differentiation, apoptosis, angiogenesis, migration, and self-renewal. Additionally, NICD activates the transcription of target genes Hes1 and Hes5 by translocating to the nucleus and forming a stable complex with RBP-J and MAML, which act as DNA-binding proteins. Therefore, compounds that can inhibit various functions of the Notch signaling pathway have the potential to become drugs for a variety of diseases involving these functions. A series of therapeutic strategies targeting the abnormalities of the Notch signaling pathway are currently under investigation and development. For instance, inhibitors of the Notch signaling pathway, such as γ-secretase inhibitors, are being studied and clinically trialed as potential treatment methods for acute lymphoblastic leukemia (ALL) and T-cell lymphoma (T-ALL). Based on the current state of Notch pathway inhibitors in clinical stages, the field is not yet crowded and is at a stage of interest to MNC companies. Eli Lilly and BMS show a high level of interest in this area. Eli Lilly has two molecules in clinical development: the Phase I LY3039478 and the discontinued LY900009. Eli Lilly's recent acquisition may also be related to collaboration on these pathway inhibitors. After all, PRISM BioLab's recent patents include three that belong to Notch inhibitors, with numerous examples in the patents and key molecules featuring more than three chiral centers. The most recently published patent, WO2023200017, has evolved from a bicyclic to a tricyclic heptamer, indicating that the synthesized molecules are becoming increasingly complex, which also demonstrates PRISM BioLab's significant focus on the Notch pathway. At present, drugs targeting the Notch signaling pathway are still under research. The Notch signaling pathway plays a crucial role in a variety of physiological processes, and intervention must be approached with caution to avoid severe toxic side effects. The regulatory mechanism of the Notch signaling pathway is complex, and how to precisely control it to achieve therapeutic effects remains an important direction for future research. How to obtain the latest research advancements in the field of biopharmaceuticals? In the Synapse database, you can keep abreast of the latest research and development advances in drugs, targets, indications, organizations, etc., anywhere and anytime, on a daily or weekly basis. Click on the image below to embark on a brand new journey of drug discovery!

▎药明康德内容团队编辑   KRAS靶向疗法的开发一直是癌症领域的焦点。在实体瘤中，KRAS G12D是最常见的KRAS突变亚型，尤其在胰腺癌（PDAC）、结直肠癌（CRC）和肺腺癌（LUAD）中频发。然而目前，特异性靶向KRAS G12D突变蛋白的抑制剂均处于临床前/早期临床研究阶段，临床上仍缺乏有效的靶向策略。 日前，《细胞》子刊《癌细胞》（Cancer Cell）上线了有关KRAS G12D特异性抑制剂开发的一项最新进展。同济大学附属上海市肺科医院任胜祥教授、周彩存教授团队，复旦大学基础医学院李飞团队，与恒瑞公司药物研发团队合作，提供了KRAS G12D抑制剂HRS-4642单药及其与蛋白酶体抑制剂联合的治疗新策略。论文指出，这是迄今为止首个被报道临床有效的KRAS G12D特异性抑制剂。 研究指出，与临床已有获批靶向抑制剂的KRAS G12C突变亚型不同，KRAS G12D缺乏靠近switch-Ⅱ结合口袋的活性残基，使得开发共价抑制剂极具挑战性。此外，为了克服KRAS抑制剂常见的耐药问题，还需联合治疗以增强疗效，因而寻找KRAS G12D抑制剂的协同增敏靶标也非常重要。 在这项研究中，作者团队研发的HRS-4642是一款高亲和力、选择性、长时效、非共价的KRAS G12D抑制剂。 体外实验表明，HRS-4642特异性结合KRAS G12D，其亲和力为KRAS G12C的21倍，野生型KRAS蛋白的17倍。结合抑制实验显示，HRS-4642抑制KRAS G12D与SOS1或RAF1结合，发挥双重阻滞作用，进而抑制下游MEK-ERK信号通路。在16个人源细胞系中，HRS-4642特异性抑制KRAS G12D突变细胞系增殖（IC50值为0.55-66.58 nM之间），而对其他KRAS突变亚型或野生型细胞系作用极弱（IC50值为248.50到大于10000 nM）。 体内实验部分，通过构建AsPC-1、GP2d异种移植瘤模型及肺腺癌PDX模型，确认HRS-4642显著抑制KRAS G12D肿瘤生长，且具有较好的药代动力学（PK）、药效学（PD）特征，倾向于瘤内蓄积。 更为重要的是，在其1期临床研究（NCT05533463）中，HRS-4642也展示出良好的抗肿瘤活性。论文显示，2名接受过多线标准抗肿瘤治疗的晚期非小细胞肺癌（NSCLC）患者，分别接受200 mg和300 mg HRS-4642（每周一次）治疗后，靶病灶分别缩小53%和31%，达部分缓解（PR）标准。这使得HRS-4642成为首个被报道临床有效的KRAS G12D特异性抑制剂。 研究团队指出：“2名经过多线标准治疗患者经HRS-4642治疗取得部分缓解，肿瘤生长受到显著抑制，充分说明KRAS G12D具有临床可靶向性，为KRAS G12D肿瘤患者提供了有效的治疗策略，标志着KRAS G12D进入靶向治疗新阶段。” ▲在参与1期临床试验的2名KRAS G12D突变肿瘤患者中，HRS-4642展示出临床活性（图片来源：参考资料[1]） 随后，为寻找HRS-4642协同增敏靶标，研究团队进一步通过体外CRISPR全基因筛选，绘制了其增敏/耐受图谱。对筛选数据进行富集分析，提示蛋白酶体或可为HRS-4642增敏靶标。对癌症基因图谱（TCGA）数据的分析也发现，蛋白酶体特征低表达与泛癌、肺腺癌及胰腺癌长生存相关。 研究团队使用蛋白酶体抑制剂卡非佐米（carfilzomib）及HRS-4642进行联合给药，发现该方案可在体外协同杀伤KRAS G12D突变细胞系，体内联合抑制KRAS G12D肿瘤生长。RNA-seq、DIA蛋白质谱等手段揭示，HRS-4642与卡非佐米联合治疗主要通过下调Notch4信号通路及上调IFNα信号通路发挥协同抗肿瘤作用。 此外，流式细胞术、免疫组化等方法还发现，HRS-4642单药或联合卡非佐米，都可重塑肿瘤微环境，显著促进KRAS G12D肿瘤中CD3+、CD4+、CD8+ T细胞的浸润及激活，促进肿瘤免疫微环境“炎化”，为KRAS G12D肿瘤“靶免联合”提供了理论基础。 “我们的研究展示了HRS-4642在多个KRAS G12D突变癌种中显著的临床前抗肿瘤活性，随着临床研究的深入进行，我们期待看到HRS-4642在其他癌肿中的疗效，”对于HRS-4642的前景，研究团队展望道，“此外，我们相信基于靶向的联合治疗策略对攻克KRAS G12D突变肿瘤尤为重要。本研究为HRS-4642联合蛋白酶体抑制剂或免疫检查点抑制剂提供了有力临床前证据，未来我们将继续探索这些联合策略临床应用的可能性，有望为KRAS G12D突变实体瘤患者提供有效、安全的靶向联合策略。” ▲研究总结示意图（图片来源：参考资料[1]） 同济大学附属上海市肺科医院李重阳博士后、罗丽波博士、贾科毅博士及上海恒瑞医药有限公司李心博士为本研究共同第一作者。上海恒瑞医药有限公司张喆博士和同济大学附属上海市肺科医院蒋涛副研究员为本研究共同通讯作者。 参考资料： [1] Zhou et al., Anti-tumor efficacy of HRS-4642 and its potential combination with proteasome inhibition in KRAS G12D-mutant cancer, Cancer Cell (2024), https://doi.org/10.1016/j.ccell.2024.06.001  本文来自药明康德内容微信团队，欢迎转发到朋友圈，谢绝转载到其他平台。如有开设白名单需求，请在“学术经纬”公众号主页回复“转载”获取转载须知。其他合作需求，请联系wuxi_media@wuxiapptec.com。 免责声明：药明康德内容团队专注介绍全球生物医药健康研究进展。本文仅作信息交流之目的，文中观点不代表药明康德立场，亦不代表药明康德支持或反对文中观点。本文也不是治疗方案推荐。如需获得治疗方案指导，请前往正规医院就诊。

引言 KRAS G12D是实体瘤中最常见KRAS突变亚型，在胰腺癌（PDAC）和结直肠癌（CRC）中排第1，在肺腺癌（LUAD）中排第2【1】。与KRAS G12C癌蛋白不同，KRAS G12D缺乏靠近switch-Ⅱ结合口袋的活性残基，使得开发共价抑制剂极具挑战性【1-3】。目前，一些研究团队基于不同策略研发了数个KRAS G12D抑制剂。例如，非共价抑制剂MRTX1133可结合switch-Ⅱ口袋，抑制KRAS G12D核苷酸交换及下游效应分子结合，从而发挥抗肿瘤效果【4,5】。其他抑制剂，如RMC-9805【6】、TH-Z835【7】及BI-2852【8】等也处于研究当中。然而，这些药物仍处于临床前/初步临床研究阶段，临床上KRAS G12D仍缺乏有效靶向策略。 此外，KRAS抑制剂面临的另一重大挑战为不可避免的耐受【9-12】。临床上，KRAS G12C抑制剂单药治疗客观有效率（ORR）为7.1%-53.4%，中位无进展生存期（PFS）为4.0-13.1个月【13-20】，亟需联合治疗以增强疗效、增加获益人群。对此，Yaeger【17】和Hallin【4】等分别发现KRAS G12C和KRAS G12D抑制剂联合西妥昔单抗可增强疗效。这些研究充分说明寻找KRAS G12D抑制剂协同增敏靶标的重要性。 2024年6月27日，同济大学附属上海市肺科医院任胜祥、周彩存教授团队与复旦大学基础医学院李飞团队及恒瑞公司药物研发团队合作在Cancer Cell上发表题为Anti-tumor efficacy of HRS-4642 and its potential combination with proteasome inhibition in KRAS G12D-mutant cancer的文章，提供了首个临床有效KRAS G12D特异性抑制剂——HRS-4642，及其与蛋白酶体抑制剂联合的治疗新策略。 在这项研究中，作者团队研发了一款高亲和力、选择性、长时效、非共价的KRAS G12D抑制剂，HRS-4642。体外，研究团队首先通过SPR实验确认HRS-4642特异性结合KRAS G12D，其亲和力为KRAS G12C的21倍，野生型KRAS蛋白的17倍。随后通过结合抑制实验，发现HRS-4642抑制KRAS G12D与SOS1或RAF1结合，发挥双重阻滞作用，进而抑制下游MEK-ERK信号通路。在16个人源细胞系中，HRS-4642特异性抑制KRAS G12D突变细胞系增殖（IC50值为0.55-66.58 nM之间），而对其他KRAS突变亚型或野生型细胞系作用极弱（IC50值为248.50到大于10,000 nM）。体内，通过构建AsPC-1、GP2d异种移植瘤模型及肺腺癌PDX模型，确认HRS-4642显著抑制KRAS G12D肿瘤生长，且具有较好的药代动力学（PK）、药效学（PD）特征，倾向于瘤内蓄积。在其I期临床研究（NCT05533463）中，HRS-4642也展示出临床活性。2名接受过多线标准抗肿瘤治疗的晚期非小细胞肺癌（NSCLC）患者，分别接受200mg和300mg HRS-4642（每周一次）治疗后，靶病灶分别缩小53%和31%，达部分缓解（PR）标准。这使得HRS-4642成为第一个被报道临床有效的KRAS G12D特异性抑制剂。 研究团队进一步通过体外全基因组CRISPR筛选，绘制了HRS-4642的增敏/耐受图谱。对筛选数据进行富集分析，确定蛋白酶体或可为HRS-4642增敏靶标。对TCGA数据进行分析也发现，蛋白酶体signature低表达与泛癌、肺腺癌及胰腺癌长生存相关。团队进而通过体内外联合给药实验，明确了HRS-4642联合蛋白酶体抑制剂卡非佐米在体外协同杀伤KRAS G12D突变细胞系，体内联合抑制KRAS G12D肿瘤生长。并通过RNA-seq、DIA蛋白质谱等手段，发现HRS-4642与卡非佐米联合治疗主要通过下调Notch4信号通路及上调IFNα信号通路发挥协同抗肿瘤作用。 除了提供HRS-4642靶向及其联合蛋白酶体抑制剂治疗新策略，研究团队通过流式细胞术、免疫组化等还发现，HRS-4642单药或联合卡非佐米可显著促进KRAS G12D肿瘤中CD3+、CD4+、CD8+ T细胞的浸润及激活，促进肿瘤免疫微环境（TIME）“炎化”，为KRAS G12D肿瘤靶免联合提供了理论基础。 模式图（Credit: Cancer Cell） 参考文献 Zeissig, M.N., Ashwood, L.M., Kondrashova, O., and Sutherland, K.D. 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Med. 389, 710–721. https://doi.org/10.1016/j.ccell.2024.06.001 责编|探索君 排版|探索君 文章来源|“BioArt” End 往期精选 围观 一文读透细胞死亡(Cell Death) | 24年Cell重磅综述（长文收藏版） 热文 Cell | 是什么决定了细胞的大小？ 热文 Nature | 2024年值得关注的七项技术 热文 Nature | 自身免疫性疾病能被治愈吗？科学家们终于看到了希望 热文 CRISPR技术进化史 | 24年Cell综述