LIG3

BEIJING--(BUSINESS WIRE)-- InnoCare Pharma (HKEX: 09969; SSE: 688428), a leading biopharmaceutical company focusing on the treatment of cancer and autoimmune diseases, announced today the approval of the Investigational New Drug (IND) to conduct the clinical trial of B-cell lymphoma-2 (BCL2) inhibitor ICP-248 in combination with Bruton's tyrosine kinase (BTK) inhibitor orelabrutinib as first-line therapy for chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) in China. This multicenter, randomized-controlled, open-label clinical study is designed to evaluate the efficacy and safety of ICP-248 combined with orelabrutinib versus immunochemotherapy in treatment-naive patients with CLL/SLL. ICP-248 is a novel, orally bioavailable BCL2-selective inhibitor, which aims to treat hematologic malignancies as a monotherapy or in combination with other therapies. BCL2 is an important regulatory protein of apoptosis pathway, and its abnormal expression is related to the development of various hematologic malignancies. ICP-248 has an anti-tumor effect by selectively inhibiting BCL2 and restoring the mechanism of programmed cell death. The preliminary result has demonstrated a good efficacy and safety profile. Orelabrutinib has been approved for marketing in China and Singapore, and all three approved indications related to lymphoma have been included in the National Reimbursement Drug List (NRDL) in China. In the treatment of relapsed/refractor CLL/SLL patients with orelabrutinib, the overall response rate and complete response rate reached 93.8% and 30% respectively, demonstrating an outstanding efficacy and safety profile. Dr. Jasmine Cui, the co-founder, chairwoman and CEO of InnoCare, said, "InnoCare has developed strong pipeline in hemato-oncology that covers a variety of important hemato-oncology targets, including BTK, BLC2, CD19, CD20xCD3, E-3 ligase, etc. ICP-248 and orelabrutinib are important global assets of our company in the field of hematology, with multiple clinical studies conducted in China and the United States. We will accelerate clinical development and look forward to providing CLL/SLL patients with more efficient and safe treatment options." CLL/SLL, one of the most common types of leukemia, is an indolent malignancy of B lymphocytes. There are 191,000 newly diagnosed CLL cases and 61,000 deaths every year globally1. The incidence rate of CLL/SLL is on the rise in China2. About InnoCare InnoCare is a commercial stage biopharmaceutical company committed to discovering, developing, and commercializing first-in-class and/or best-in-class drugs for the treatment of cancers and autoimmune diseases with unmet medical needs in China and worldwide. InnoCare has branches in Beijing, Nanjing, Shanghai, Guangzhou, Hong Kong, and the United States. InnoCare Forward-looking Statements This report contains the disclosure of some forward-looking statements. Except for statements of facts, all other statements can be regarded as forward-looking statements, that is, about our or our management's intentions, plans, beliefs, or expectations that will or may occur in the future. Such statements are assumptions and estimates made by our management based on its experience and knowledge of historical trends, current conditions, expected future development and other related factors. This forward-looking statement does not guarantee future performance, and actual results, development and business decisions may not match the expectations of the forward-looking statement. Our forward-looking statements are also subject to a large number of risks and uncertainties, which may affect our short-term and long-term performance. _________________________ 1 American Journal of Hematology 2 Ybanlv

BEIJING--(BUSINESS WIRE)-- InnoCare Pharma (HKEX: 09969; SSE: 688428), a commercial-stage biotech company, announced today that the U.S. Food and Drug Administration (FDA) has cleared the Investigational New Drug (IND) application for the Company’s B-cell lymphoma-2 (BCL2) inhibitor, ICP-248. This is InnoCare’s fifth innovative drug to enter the clinical stage in the U.S. This is a Phase I study to evaluate the safety, tolerability, pharmacokinetics and preliminary efficacy of ICP-248 in hematologic malignancy patients. ICP-248 is a novel, orally bioavailable BCL2-selective inhibitor, which aims to treat hematologic malignancies as a monotherapy or in combination with other therapies. The Phase I dose escalation trial of ICP-248 is ongoing in China, and the preliminary results demonstrated good efficacy and safety pro BCL2 is an important regulatory protein of the apoptosis pathway, and its abnormal expression is related to the development of various hematologic malignancies. ICP-248 exhibits its anti-tumor effects by selectively inhibiting BCL2 and restoring the mechanism of programmed cell death. Dr. Jasmine Cui, the Co-founder, Chairwoman and CEO of InnoCare said, “ICP-248 would become an important asset for the Company’s globalization after orelabrutinib. The current study results will support ICP-248 to treat hematologic malignancies as a monotherapy or in combination with other therapies. Meanwhile, InnoCare is dedicated to building a leading franchise in hemato-oncology, and we have developed multiple drugs that cover a variety of important hemato-oncology targets such as BTK, CD19, CD20xCD3, BCL2 and E-3 ligase to address unmet medical needs.” About InnoCare InnoCare is a commercial stage biopharmaceutical company committed to discovering, developing, and commercializing first-in-class and/or best-in-class drugs for the treatment of cancers and autoimmune diseases with unmet medical needs in China and worldwide. InnoCare has branches in Beijing, Nanjing, Shanghai, Guangzhou, Hong Kong, and the United States. InnoCare Forward-looking Statements This report contains the disclosure of some forward-looking statements. Except for statements of facts, all other statements can be regarded as forward-looking statements, that is, about our or our management's intentions, plans, beliefs, or expectations that will or may occur in the future. Such statements are assumptions and estimates made by our management based on its experience and knowledge of historical trends, current conditions, expected future development and other related factors. This forward-looking statement does not guarantee future performance, and actual results, development and business decisions may not match the expectations of the forward-looking statement. Our forward-looking statements are also subject to a large number of risks and uncertainties, which may affect our short-term and long-term performance. View source version on businesswire.com: Contacts Media Chunhua Lu 86-10-66609879 chunhua.lu@innocarepharma.com Investors 86-10-66609999 ir@innocarepharma.com Source: InnoCare Pharma View this news release online at:

1DNA双链断裂（DSB）修复途径合成致死性是诱导细胞死亡的生物过程，其基于同时抑制在细胞存活所需的过程中的两种途径并行作用，仅抑制一种途径会导致细胞存活。合成致死策略已在抗癌治疗中得到广泛应用。由于一种途径可能由于转化相关变化而在癌细胞中失活，因此靶向另一种途径触发细胞死亡，同时保留健康细胞1。癌细胞的关键特征之一是由DNA损伤的积累，包括DNA双链断裂（DSB），这是细胞中最致命的DNA损伤之一。然而，癌细胞能够通过调节其DNA修复途径存活和增殖，这可能与正常细胞中的DNA修复途径不同1。DSB可以通过两种主要机制进行修复：BRCA1/2介导的同源重组（HR）和典型 DNA-PKcs介导的非同源末端连接（c-NHEJ）（图1）1。HR被认为是一种准确的DSB修复途径，因为它依赖于细胞周期S/G2期细胞的姐妹染色单体作为DNA合成和DSB修复的模板。因此，HR能够修复DSB，主要在增殖细胞中1。HR修复途径由两个经典的肿瘤抑制基因BRCA1和BRCA2编码的蛋白质进行全面调节，然后募集RAD51及其旁系同源物（包括RAD51B，RAD51C，RAD51D，XRCC2和XRCC3）和RAD54以识别同源DNA模板并进行链侵袭以修复DSBs。另一方面，NHEJ是一种容易出错的DSB修复机制。NHEJ可以是分为c-NHEJ和a-NHEJ（也称为MMEJ或TMEJ）两种途径1。图1. DNA双链断裂（DSB）修复途径包括同源重组（HR），规范非同源末端连接（c-NHEJ）和替代非同源末端连接（a-NHEJ）/微同源介导的末端连接（MMEJ）/DNA聚合酶θ（Polθ）介导的末端连接（TMEJ）2PARPi在造血恶性肿瘤中的治疗潜力最近的很多研究表明，即使在白血病中很少检测到BRCA1/2突变，PARP抑制剂（PARPi）诱导的合成致死性也可以有效地在 BRCA1/2 缺陷造血恶性细胞中起作用。此外，诱导造血恶性肿瘤的遗传改变，例如驱动骨髓和淋巴恶性肿瘤的癌基因，包括 AML1-ETO（也称为 RUNX1-RUNX1T1）、BCR-ABL1、PML-RARa、TCF3-HLF、IDH1/2mut和IGH-MYC以及肿瘤抑制基因（例如TET2、WT1）的功能缺失突变可导致HR和/或c-NHEJ活性失调，从而使细胞容易受到PARPi引发的合成致死效应的影响（图2和图3）1。图2. PARP抑制剂在造血恶性肿瘤和其他显示癌基因/突变（A）和酪氨酸激酶抑制剂（B）诱导的HR/c-NHEJ缺乏症的肿瘤中的应用此外，Bac等人描述了那些恶性造血细胞表达致癌酪氨酸激酶 （OTK）（例如 BCR-ABL1、FLT3（ITD）、JAK2（V617F））由于ROS产量增加自发积累高水平的氧化DNA损伤和DSB 1。然而，OTK阳性细胞能够逃逸来自 DSB 的细胞毒性作用，这是由于增强/调节的 DSB 修复。FDA批准的特异性酪氨酸激酶抑制剂（TKi）（JAK1/2抑制剂鲁索利替尼，FLT3抑制剂奎扎替尼，ABL1抑制剂伊马替尼）对这些OTK的抑制导致急性HR / c-NHEJ缺乏症（由于BRCA1，BRCA2，RAD51和/或LIG4的下调）和对PARPi的敏感性（图2-4）。因此，TKi和PARPi的联合使用能够根除增殖和静止的恶性造血干细胞和祖细胞1。图3. 诱发HR / c-NHEJ缺乏症的癌基因/突变图4. 诱导HR/c-NHEJ缺乏的治疗药物3对PARPi的获得性耐药机制尽管 PARPi 在 DSB 修复缺陷的癌细胞中具有很强的效力，但在BRCA1/2 缺陷和 HRD 中已经报道了对 PARPi 诱导的合成致死性的抵抗力肿瘤细胞（图5）。PARPi的耐药性的主要原因是BRCA1/2突变癌细胞HR修复活性的恢复2。详细地说，BRCA1/2中的继发突变与原始突变导致的链终止子/移码的废除有关，导致全长 BRCA1/2 开放阅读框的恢复。因此，活性BRCA1/BRCA2蛋白表达被恢复，从而重新启动功能齐全的HR途径3,4。除了BRCA1/2中其他突变介导的耐药性外，还提出了BRCA1启动子甲基化的减少/丧失，从而恢复了BRCA1的表达而导致对PARPi的耐药性5。图5. 对PARPi的获得性耐药机制BRCA1-BRCA2-RAD51轴对于同源重组修复（HRR）至关重要。EMI1调节三阴性乳腺癌（TNBC）细胞中的 PARPi 敏感性。EMI1的F盒结构域组装一个SCF泛素连接酶复合物，该复合物靶向RAD51进行降解。在一部分对PARPi难治的BRCA1缺陷三阴性乳腺癌细胞中，EMI1表达的降低会损害EMI依赖性RAD51的降解，从而恢复HR修复活性导致对PARPi的耐药性（图6）6。图6. KEAP1的增加导致EMI1表达的降低导致对PARPi的耐药机制EMSY是I型干扰素反应的负调节因子，也抵消了HR修复途径，KEAP1靶向EMSY进行泛素介导的降解。因此，KEAP1的过表达破坏了EMSY的稳定性，导致HR恢复和对PARPi的抵抗。相反，由于基因突变引起的KEAP1失活导致EMSY的稳定和HR缺乏而使得非小细胞肺癌对PARPi变得敏感（图7）7。图7. EMI1表达的降低导致EMSY的减少，从而对PARPi的耐药机制PARP1的下调或者突变都可能导致BRCA1/2缺陷肿瘤细胞中的PARPi耐药性（图5）。通过53BP1-RIF1的复合物，53BP1介导的DSB末端切除术抑制导致c-NHEJ活性增强，减少对PARP1依赖性a-NHEJ的依赖性，从而引起PARPi耐药性8.因此，53BP1在BRCA1-null小鼠胚胎干细胞中的抑制促进DSB末端切除以限制c-NHEJ，从而使细胞对PARPi致敏。在BRCA1突变的乳腺癌细胞中，由于HR修复活性的部分恢复，53BP1的体细胞丢失导致奥拉帕尼耐药，从而降低BRCA1缺陷小鼠乳腺肿瘤对该抑制剂的反应（图5）。53BP1定位于DSB的阻断也有助于BRCA1缺陷肿瘤中PARPi的失活，这是由TIRR表达增加介导的9。SHLD1/2/3复合物的失活以及DSB末端切除术中其他蛋白（包括RIF1和REV7）的表达降低，恢复了HR的功能活性，降低了PARPi的疗效（图8）10。以相同的机制方式，TRIP13是SHLD1 /2 /3复合物的负调节因子，通过解离REV7-SHLD1/2/3去促进HR。因此，TRIP13的扩增诱导BRCA1/2突变癌症对PARPi的耐药11。图8. SHLD1/2的敲除在BRCA1敲除细胞中导致对PARPi的耐药机制除了在HR通路中起作用外，据报道，BRCA1和BRCA2在维持复制分叉的完整性方面发挥作用12。在BRCA1和BRCA2缺陷细胞中对PARPi获得性耐药可能是由于EZH2缺失导致MRE1和MUS81核酸酶表达降低，导致对复制叉的保护性反应（图9）12,13。图9. MRE1、MUS81和EZH2的下调导致对PARPi的耐药机制除了基于恢复HR或复制叉保护的获得性PARPi抗性机制之外，最近的一项研究报告称，通过恢复Okazaki片段处理（OFP）减少/丢失DNA复制间隙可以诱导抗性。基本上，据报道，复制间隙是BRCA1/2缺陷细胞中PARPi反应的主要决定因素。一般来说，BRCA1/2的缺乏会扩大复制间隙，导致细胞的PARPi敏感性1。然而，在同时敲除BRCA1和53BP1的细胞模型中，通过恢复XRCC1-LIG3恢复OFP，导致减少/耗尽复制差距，并最终对PARPi产生抵抗力。相反，双倍敲除53BP1和LIG3使BRCA1突变细胞对PARPi重新敏感（图10）14。图10. XRCC1-LIG3的增加恢复OFP对PARPi产生耐药性4预先存在的 PARPi 耐药性机制HR和/或c-NHEJ缺陷的血液系统恶性肿瘤在模拟外周血微环境（PBM） 的条件下对PARPi介导的合成致死性敏感，在模拟骨髓微环境（BMM）的条件对 PARPi 产生抗性。Bac等人表明转化生长因子β1（TGF-1）由骨髓基质细胞产生的激活缺氧诱导的TGF-受体（TGF R）激酶-SMAD2/3在白血病细胞中的规范途径以促进DSB的修复（图11）1。TGFβR激酶抑制剂没有改变白血病细胞对PBM中PARPi的敏感性，表明DSB修复活动中TGFβR-SMAD3信号传导的调节在BMM中是排他性的。在机制上，TGFβR激酶信号通过BRCA1/2、ATM，DNA-PKcs和LIG4的上调刺激BMM中白血病细胞中DSB的修复。这一发现表明TGFβR激酶抑制剂在PARPi介导的造血恶性肿瘤干预中的潜在临床应用。事实上，TGFβR激酶的抑制剂galunisertib已经在临床上几种癌症的试验得到应用 15,16，恢复了BMM中白血病细胞的PARPi敏感性。因此，Bac等人假设在PARPi治疗中加入galunisertib应该改善治疗效果1。图11. 通过在缺氧骨髓微环境中激活TGF-β1—TGFβR—SMAD2/3信号传导，恢复HR/c-NHEJ缺陷白血病中的HR/c-NHEJ的PARP抑制剂耐药的预先存在机制除此之外，还有一些其它预先存在的对PARPi的耐药机制，比如DNMT3A的突变导致功能丧失，FLT3激酶与ITD融合突变导致FLT3激酶激活，JAK2的V617K突变导致JAK2激酶的激活，c-KIT的N822K突变导致c-KIT激酶的激活，EGFR的激活和IGF-1R的激酶等都可能导致预先存在的PARPi抑制剂的耐药（图12）1。图12. 预先存在的 PARPi 耐药性机制5小结鉴于负责PARPi耐药性的机制途径列表不断增加，必须实现对预先存在和获得的PARPi耐药的药理学抑制，以提高PARPi介导的合成的治疗效率杀伤力。例如，白血病细胞中预先存在的PARPi耐药性可以通过联合PARP和TGFβR激酶抑制剂（使BMM中的白血病细胞再敏化）、PARP和TET2双加氧酶抑制剂（使携带DNMT3A突变的白血病重新致敏）以及PARP和OTK抑制剂（FLT3（ITD）、JAK2（V617F）、c-KIT（N822K）、EGFR、IGF-1R）解决。获得性 PARPi 抗性可通过以下方式预防，如更具侵略性的“双重合成致死”方法（同时靶向PARP1和RAD52），可在更短的时间内消除更多的肿瘤细胞，从而减少抗性克隆出现的时间依赖性的机会。此外，如果 PARPi出现耐药性，这些克隆可以通过抑制另一种DNA修复机制来消除，例如Pol介导的TMEJ 17。参考文献：1.Bac Viet Le, Paulina Podszywałow-Bartnicka, Katarzyna Piwocka, and Tomasz Skorski, Pre-Existing and Acquired Resistance to PARP Inhibitor-Induced Synthetic Lethality, Cancers 2022, 14, 5795.2.Quigley, D.; Alumkal, J.J.; Wyatt, A.W.; Kothari, V.; Foye, A.; Lloyd, P.; Aggarwal, R.; Kim, W.; Lu, E.; Schwartzman, J.; et al. 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