CFT-1946

学以致用   侃侃而言 RSC Med Chem 2024, Nov. 7th DOI: 10.1039/D4MD00769G 蛋白质水解靶向嵌合体（PRTOTACs）是一种靶向蛋白降解的新兴治疗策略，它们将靶标蛋白（POI）与E3连接酶紧密结合，并诱导参与泛素-蛋白酶体系统（UPS）以促进靶向蛋白质降解。 近日，RSC Med Chem上刊载了一篇总结性研究，通过分析处于临床中的PROTACs实体分子的物理化学性质，汇总了PROTACs各组成部分的优化，以及对于实现口服生物利用度所面临的困难和挑战，并根据作者在这一领域的经验提出了一些珍贵的指导性意见和建议。 主要内容 (1) 临床中的PROTACs (2) 口服生物利用度开发中的挑战 (3) 值得注意的陷阱提示 (4) 总结与展望，及往期相关精彩回顾 01 临床中的PROTACs PROTACs等靶向降解剂是一种“事件驱动”的治疗策略，其选择性地从细胞中去除靶标蛋白质。相比于传统的基于“占位驱动”的蛋白质功能抑制策略，PROTACs等靶向降解剂具有几个优势： (1)药理学上，PROTACs等靶向降解剂有可能提供更加完整的蛋白质功能封锁（如去除支架活性），因此，更有可能概括基因敲除实验的效果。简单地说，PROTACs等靶向降解剂不仅阻断了信号传导，而且通过分解细胞传播信号所需的机制，消除了细胞传播信号的能力； (2)细胞只能通过产生新的蛋白质来恢复功能，因此，药理学受到蛋白质的再合成率以及药物药效学的影响，从而提供了延长作用时间的可能性； (3) PROTAc机制中POI与E3组分之间必要的相互作用，为异构体之间（如，取决于三元复合物的形成和/或可用的表面赖氨酸残基）和组织内（如，取决于E3表达水平）的选择性提供了额外的机会； 因此，这种药物实体已经出现，便被证明对于患者具有良好的耐受性和有效性，并已有多个实体分子进入后期临床研究。 据统计，截至2024年7月，已有13种结构已公开的PROTACs进入临床试验阶段。这些PROTACs大多数用于肿瘤相关适应症，其中一种用于治疗炎症；两种因战略规划原因而停止（ARV-110、KT-413）。 就E3连接酶配体而言，其中有12种携带CRBN配体，口服给药。而仅有一种需要静脉注射（IV）的DT-2216，携带VHL配体。如下图。 图片源自RSC Med Chem 这些处于临床阶段的PROTACs实体的物化性质统计如下表。其中，携带CRBN配体的PROTACs分子量在711~958 Da；亲脂性（pH7.4时的LogD值）为1.4~4.8；色谱极性表面积（ePSA）为107~146 Å 2。 图片源自RSC Med Chem 而计算出的分子亲脂性LogP和极性表面积tPSA的预测结果分散更为广阔：其中LogP为-0.6~6.8；tPSA范围为96~238 Å 2。此外氢键受体HBA为9~20；氢键供体HBD均≤5。 如下图A，通过雷达图更形象地展示了临床口服PROTACs的物化性质空间（蓝色区域）在所有特性方面都超出了Lipinski五规则（Ro5）和Veber关于可旋转键（RotB）和极性表面积（PSA）（绿色区域）的经验指南，仅HBD除外。 图片源自RSC Med Chem 上图B绿色条形图显示了小分子的可接受的性质范围，同时与临床PROTACs分子做了对比。 在亲脂性方面，一些PROTACs突破了小分子LogP（≤5）和LogD（1~3）的极限。值得注意的是，ePSA的值（107~141 Å2）比tPSA （96~238 Å2）的范围窄得多，这表明这可能是口服PROTACs设计的更有价值的指南。尽管超出了先前确定的物理化学空间限制，口服给药仍然是首选的给药途径，专门为血液病癌症开发的DT2216和KT-413除外。 以上每一种PROTACs都是经历了广泛的优化后，以实现药理学和物化性质的最佳平衡的结果。而PROTACs模块化、三组件的特性（POI配体、E3配体、连接子），其设计的技巧在于平衡单个部件组分的特征，从而使最终PROTACs的整体特性处于可接受的空间内。 就POI配体而言：强效，较少的暴露HBD 许多PROTACs针对靶标POI改良的抑制剂/拮抗剂进行设计。这些抑制剂或拮抗剂通常是高效的配体片段基序，在其溶剂区安装输出载体，用以连接到E3结合基序片段。此外，这些配体大多数是中性的，而非酸性或碱性，并且氢键供体HBD≤1。以上几个PROTACs中也有几个例外，如ARV-393、NX-2127、KT-413、NX-5948，整体分子分别包含3、4、4、5个HBD，但分子内氢键可能屏蔽了其中的几个HBD。 就E3连接酶配体而言：CRBN配体最流行 在已经揭示了分子结构的临床PROTACs中，上面已提到，除了DT-2216以外，均是携带了CRBN配体。如下图，使用最广泛的CRBN配体包括来自免疫调剂亚胺类药物（IMiDs）沙利度胺、和来那度胺。这两者均具有高级性、高效、低分子量、并且在基本结合药效团中只有一个HBD。此外，它们对外排转运体缺乏敏感性，使其成为PROTACs的理想选择，特别是在实现口服生物利用度上。 图片源自RSC Med Chem 而与IMiD类配体相关的安全性问题（如致畸性、血液毒性等），也可以解释为什么PROTACs最初的治疗适应症主要是肿瘤领域。也许正是因为这一点，当前正在出现不同于IMiD类的由新型CRBN配体组成的PROTACs。 就连接体而言：胺基碱及富含环型基序片段占主导 除了极个别例外，如CFT-1946，这些临床阶段的PROTACs的连接体的共同特征是存在一个叔胺碱，以及哌啶、哌嗪、或氮杂啶都行。在一些特定的情况下，哌啶的碱度似乎随着氟取代（CFT-8634）或氧取代（ARV-393、KT-474）的加入而有所缓和，使其碱度不超过所有口服PROTACs的预测值pKa=9.1。 而静脉给药的PROTACs，如DT-2216，具有非常高的预测亲脂性（LogP=15.0）；然而，这降低到低得多的LogD=3.9，可能是由于其两性离子电荷所致；其高试验极性为193 Å2，比其他口服PROTACs的ePSA值高出52 Å2，比相关bRo5口服药物venetoclax的ePSA值高出31 Å2，似乎反映了这种两性离子。 口服PROTACs的另一个共同之处是使用环状基序作为构象约束，所有实例在连接体中至少包含两个环，有些甚至三个环。 在灵活性方面，10个实例在环之间包含亚甲基作为间隔剂，只有BMS986365具有乙氧基3-原子间隔剂。此外，CFT-1946具有最刚性的连接体，以酰胺键连接哌啶和螺环。而KT-474具有最大的非环组分，具有4-原子丙炔醚间隔，但即使这样也相对刚性，这表明构象约束是PROTACs的重要设计考虑因素。 一些研究也表明富环的、刚性连接体可以产生更高的细胞靶标降解能力和更高的代谢稳定性。也因此，可旋转键仅略高于Ro5依从性口服药物经验指导的上限。而其中一个明显的例外则是DT-2216，具有29个可旋转键，高度灵活的连接体仍然可以产生强效的降解物，并且适用于静脉给药的PROTACs。 02 口服生物利用度的挑战 PROTACs分子由三个组分构成的结构特征使其在物化特性和口服生物利用度等方面面临着诸多挑战。 EARLY R&D 2.1、最严格的HBD，口服PROTACs设计的关键 首先，构建分子量MW低于~700Da的PROTACs具有挑战性。 这种对Lipinski的MW经验规则的违反，通常也会伴随着维持其它Ro5标准（如HBA、HBD、LogP）和其它物化特性的限制（如，PSA、可旋转键等）。 尽管如此，如上一单元所示，也有不少临床中的PROTACs分子是可以生物利用的，尤其是在人类中达到有效的暴露水平。随着越来越多的试验数据的积累，以及对关键理化性质的分析总结，一些新的bRo5经验指导出现，并必将越来越成熟和精准。 例如，Arvinas和AstraZeneca都强调了限制口服吸收的或溶剂暴露的HBDs（eHBDs）数量的重要性。两个团队都得出这样的结论：虽然物化性质，如分子量、亲脂性、分子极性和HBA等可以超过口服药物的Ro5限制，但eHBD≤2这个标准是一个严格有效的设计标准。 对于bRo5化学空间里的大环类药物，相关研究发现，酰胺类HBD的上限为2个，而总体正式的HBD上限为7个，这相对宽松了许多，大概也是因为分子内氢键屏蔽约束的结果。 回到PROTACs来说，也或许也解释了为什么CRBN配体在临床PROTACs组成中更受青睐，因为这些基序片段小、有效且只有一个eHBD。 因此，尽管挑战重重，通过合理使用分子属性可支配限制，是设计PROTACs的关键，特别是最严格的方面，即HBD。 EARLY R&D 2.2、三元复合物本身的问题带来的挑战 尽管PROTACs能够有效地形成促进表面赖氨酸泛素化的三元复合物，然则其也仅能针对靶向蛋白质发挥降解功能。 这种三聚体复合物的建模是非常具有挑战性的。 这是因为，可能存在多种三元复合物构象，并且并非所有这些构象都能导致有效的靶标降解，这无疑阻碍了基于结构的PROTACs设计。 相比于传统小分子设计程序，PROTACs的设计要考虑分子大小、“POI-连接体-E3”组分之间的构象和物理化学性质相互作用，这意味着有更多的“化学等式”需要优化。 考虑到构成可及的三元配合物的不确定性，以及它位于正确的物化空间，因此，生产“活性”PROTACs的复杂性将会导致一系列设计和合成方面的挑战。 针对这么多不确定性，一种解决办法是通过生产“库”，用来探索广阔的化学空间，从而实际地解决以经验方式存在的巨大的组合优化。 EARLY R&D 2.3、变形嵌合体的挑战 PROTAC的三元组成和分子大小，使理解其构象比传统小分子更具挑战性。 一般，预先组织分子使其具有生物活性构象，以减少结合时的熵损补偿是一种行之有效的策略。然而，对于PROTACs分子，配体可以获得更大程度的构象自由度，再加上三元复合物的构象可塑性和不确定性，使得准确确定要模仿的“生物活性”构象具有挑战性。 利用核磁共振技术，结合分子动力学模拟来了解PROTACs的构象行为及其对渗透率的影响是目前的一种方法。 在极性溶剂和非极性溶剂中观察到的差异性，为构象集成、端到端反折叠和潜在的分子内氢键（IMHB）提供了证据。研究发现，与未折叠的配对对比，CRBN和VHL安装的PROTACs的溶剂依赖性（即“变色龙”效应）分子内折叠，增加了它们的被动细胞通透性。 EARLY R&D 2.4、PROTACs体外试验生产数据的分析 对于PROTACs领域的工作人员来说，另一个挑战是使用基于传统小分子优化程序配置的体外分析生成对PROTACs有意义的数据的能力。 如前面所提，PROTACs分子大，通常是亲脂性的碱，有粘附在实验仪器/蛋白质/膜上的倾向，导致渗透性和蛋白质结合试验回收率很低。传统的摇瓶法亲脂性分析通常难以达到LogD＞4，而且溶解度通常低于检测极限，这使得在缺乏数据的情况下筛选候选药物极具挑战性。 而且，作者还根据经验提到，代谢稳定性测定（肝细胞和微粒体孵育）产生可靠的值，在淘汰不稳定的PROTACs方面是没有价值的。 色谱法（ChromologD、ePSA）扩大了亲脂性和暴露极性的可测范围，并允许在一系列化合物中对化合物进行排序。此外，已有报道将色谱方法与计算机方法相结合来生成更多相关的3D描述符，以提高设计口服可用PROTACs的成功率。 03 陷阱提示 在整个PROTACs中整合CRBN结合实体，特别是IMiDs，会带来需要监测的相关安全风险。这些风险可能与以下有关： 与IMiDs非常相似，相应的PROTACs可以作为分子胶降解剂，并导致许多蛋白质降解，从而诱发毒理学效应；例如，值得注意的一个新的底物，GSPT1，一个平移终止因子。其的无意降解降低了蛋白质合成的速率，导致细胞毒性，同时伴有POI的明显和误导性降解。 总的来说，这些发现强调需要细致的对照实验来询问PROTACs的作用机制（MoA），从而促进其优化为有效和安全的药物。从发现阶段到体内PROTACs的评估，一个关键教训是了解它们的代谢命运的重要性。 即使对于代谢相对稳定的PROTACs，也可能产生极性更强、亲脂性更低和未结合清除率更低的片段。然后，它们可以竞争与POI或E3结合，并降低三元复合物形成的有效性，从而降低体内疗效。 此外，即使精心设计的利用CRBN的PROTAC的代谢物，没有分子胶降解剂（MGD）活性，理论上也可以作为MGD，从而引入MGD相关的安全责任。因此，在当前临床试验执行过程中对此类风险进行评估，无疑将使科学界了解利用CRBN的PROTACs用于非肿瘤适应症的适用性。 04 总结与展望 PROTACs目前已经从初始的肿瘤适应症领域向其它领域扩展，但主要还是在肿瘤领域，并且当前临床中的PROTACs还是以利用CRBN作为首选的E3连接酶。 初步数据也证实，人类的生物功效和口服生物利用度也是可以实现的。通过已公开的数据，得到的结论是它们在一些结构上（如碱性、富环连接体）和关键物化特性上（如eHBD≤2）等方面表现出了统一性的特征，并具有一定的指导意义。 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↓

Cemsidomide Phase 1/2 Trial in Multiple Myeloma and non-Hodgkin’s Lymphoma Continues to Progress: Data from Both Indications to be Presented at the 66th American Society of Hematology (ASH) Annual Meeting in San Diego, CA Initial Monotherapy CFT1946 Phase 1 Data Demonstrated Well-Tolerated Safety Profile and Early Evidence of Anti-tumor Activity; Phase 1/2 Trial Continues to Progress with Multiple Data Readouts Expected in 2025 Appointed Paige Mahaney, Ph.D. as Chief Scientific Officer Bringing More than 25 Years of Pharmaceutical and Biotech Experience Cash, Cash Equivalents and Marketable Securities of $284.4 million as of September 30, 2024; Expected to Provide Runway into 2027 WATERTOWN, Mass., Oct. 31, 2024 (GLOBE NEWSWIRE) -- C4 Therapeutics, Inc. (C4T) (Nasdaq: CCCC), a clinical-stage biopharmaceutical company dedicated to advancing targeted protein degradation science, reported financial results today for the third quarter ended September 30, 2024, as well as recent business highlights. “2024 has been a successful year for C4T marked by strong execution across our entire portfolio, which has continued to position us as a leader in targeted protein degradation science. We recently shared initial clinical data on CFT1946 demonstrating a well-tolerated safety profile and evidence of monotherapy anti-tumor activity in BRAF inhibitor exposed patients, and we delivered our second development candidate to Biogen,” said Andrew Hirsch, president and chief executive officer of C4 Therapeutics. “We look forward to continuing this momentum with cemsidomide Phase 1 data in multiple myeloma and in non-Hodgkin’s lymphoma being presented at ASH, and multiple CFT1946 data readouts expected in 2025. Our continued focus on discovery research, clinical development and collaborations sets us up for an exciting future and for the potential to improve patients’ lives.” THIRD QUARTER 2024 AND RECENT ACHIEVEMENTS Cemsidomide: Cemsidomide is an oral degrader of IKZF1/3 for the potential treatment of relapsed/refractory (R/R) multiple myeloma (MM) and R/R non-Hodgkin’s lymphoma (NHL). Advanced the Phase 1/2 Clinical Trial. The cemsidomide Phase 1/2 trial in combination with dexamethasone for R/R MM and as a monotherapy for R/R NHL continues to enroll patients. Dose escalation continues as the maximum tolerated dose has not yet been reached. For the combination with dexamethasone MM arm, patients are now enrolling at dose level 6 (100 µg once daily (QD)) and enrollment in the dose level 5 (75 µg QD) expansion cohort is complete. The monotherapy NHL arm continues to progress and dose level 5 (100 µg QD) is the highest dose level evaluated to date. CFT1946: CFT1946 is an oral degrader targeting BRAF V600 mutations for the potential treatment of solid tumors including colorectal cancer (CRC), melanoma and non-small cell lung cancer (NSCLC). Presented New Preclinical Data at the 7th Annual Targeted Protein Degradation (TPD) Summit: In October 2024, C4T shared new in vivo data for CFT1946 describing Kpu,u (range: 0.34 - 0.88), experimentally determined using independent methods in two different species. These results demonstrate CFT1946’s ability to cross the blood brain barrier and highlight the potential for activity in primary and metastatic central nervous system (CNS) disease.Presented Monotherapy Clinical Data Demonstrating Proof of Mechanism and Early Evidence of Proof of Concept from the Ongoing CFT1946 Phase 1 Trial at the European Society for Medical Oncology (ESMO) Congress 2024: CFT1946 demonstrated a well-tolerated safety profile and evidence of monotherapy anti-tumor activity, which supports further clinical development as monotherapy and in combination with MEK and EGFR targeted therapies.Advanced the Phase 1/2 Clinical Trial. The CFT1946 Phase 1/2 trial for BRAF V600 mutant solid tumors continues to enroll patients across multiple arms of the trial: CFT1946 Monotherapy Dose Escalation in BRAF V600 Mutant Solid Tumors: Patients continue to enroll in the 640 mg twice daily (BID) pharmacodynamic (PD) backfill cohort.CFT1946 Monotherapy Melanoma Expansion Cohorts: Enrollment at the 320 mg BID dose level is complete and patients are enrolling at the 640 mg BID dose level.CFT1946 with Cetuximab Phase 1b Dose Escalation for CRC: The 160 mg BID dose level has been declared safe and patients are enrolling at the 320 mg BID dose level. RESEARCH UPDATES: Presented at the 7th Annual TPD Summit Highlighting C4T’s TORPEDO® Platform: In October 2024, at the 7th Annual TPD Summit, C4T presented kinetics-based pharmacokinetic (PK) and PD models, demonstrating the company’s ability to predict clinical PD responses across target classes. COLLABORATIONS: Delivered Second Development Candidate to Biogen. In September 2024, C4T earned an additional $8 million milestone payment after Biogen accepted delivery of a second development candidate in an undisclosed indication. Biogen is responsible for all future clinical development and commercialization for this program. This marks the final development candidate under this strategic collaboration. CORPORATE UPDATES: In October 2024, C4T appointed Paige Mahaney, Ph.D., as the company’s chief scientific officer, succeeding Stew Fisher, Ph.D., who is retiring to pursue personal interests. Dr. Mahaney brings over 25 years of pharmaceutical executive leadership with multidisciplinary expertise in discovery research and development along with successfully building clinical portfolios across a wide range of disease indications and treatment modalities.In September 2024, C4T appointed Stephen Fawell, Ph.D., to the Board of Directors. Dr. Fawell brings decades of experience leading discovery and development strategies for global pharmaceutical companies. KEY UPCOMING MILESTONES Cemsidomide: Present updated dose escalation and expansion cohort data from the 50 µg M/W/F, 37.5 µg QD, 62.5 µg QD, and 75 µg QD dose levels in approximately 40 patients from the ongoing Phase 1/2 clinical trial with dexamethasone in R/R MM at the ASH Annual Meeting in December 2024.Present dose escalation data from the 25 µg M/W/F, 50 µg M/W/F, 37.5 µg QD, 62.5 µg QD, and 100 µg QD dose levels in approximately 25 patients from the ongoing monotherapy Phase 1/2 clinical trial in R/R NHL at the ASH Annual Meeting in December 2024.Complete Phase 1 dose exploration in R/R MM and R/R NHL by year-end. CFT1946: Initiate the Phase 1b dose escalation cohort evaluating CFT1946 in combination with trametinib in melanoma by year-end.Present data from multiple anticipated readouts in 2025, including: Full monotherapy CFT1946 dose escalation cohorts in BRAF V600 mutant solid tumors.Expansion cohorts evaluating CFT1946 as a monotherapy in melanoma.Phase 1b dose escalation cohort evaluating CFT1946 in combination with cetuximab in CRC. UPCOMING INVESTOR EVENTS: November 20, 2024, at 8 am GT: Management will participate in a fireside chat at the Jefferies London Healthcare conference taking place in London, UK.December 8, 2024: Management will host an investor call to discuss MM and NHL data from the ongoing Phase 1/2 trial of cemsidomide. THIRD QUARTER 2024 FINANCIAL RESULTS Revenue: Total revenue for the third quarter of 2024 was $15.4 million, compared to $11.1 million for the third quarter of 2023. The increase in revenue was primarily due to recognition of an $8.0 million milestone from Biogen as well as $2.9 million of revenue recognized under our license and supply agreement with Betta, partially offset by reduced revenue from our agreement with Roche due to the completion of research activities in the third quarter of 2023 for a nominated target. Total revenue for the third quarter of 2024 reflects revenue recognized under our collaborations with Biogen, Betta, Merck, Merck KGaA, Darmstadt, Germany (MKDG) and Roche, and total revenue recognized in the third quarter of 2023 reflects revenue recognized under collaboration agreements with Biogen and Roche. Research and Development (R&D) Expense: R&D expense for the third quarter of 2024 was $31.8 million, compared to $28.3 million for the third quarter of 2023. The increase in R&D expense was primarily due to increased clinical trial expense as cemsidomide and CFT1946 continue to advance, partially offset by lower personnel costs. General and Administrative (G&A) Expense: G&A expense was $11.8 million for the third quarter of 2024, compared to $10.5 million for the third quarter of 2023. The increase in G&A expense was primarily attributable to higher personnel expense related to stock-based compensation. Net Loss and Net Loss per Share: Net loss for the third quarter of 2024 was $24.7 million, compared to $27.0 million for the third quarter of 2023. Net loss per share for the third quarter of 2024 was $0.35 compared to $0.55 for the third quarter of 2023. Cash Position and Financial Guidance: Cash, cash equivalents and marketable securities as of September 30, 2024 were $284.4 million, compared to $295.7 million as of June 30, 2024, and $281.7 million as of December 31, 2023. The reduction in cash, cash equivalents and marketable securities during the third quarter was primarily the result of cash used in operating activities, partially offset by $10.3 million in net proceeds raised through our at-the-market (ATM) facility. C4T expects that its cash, cash equivalents and marketable securities as of September 30, 2024 will be sufficient to fund planned operating expenses and capital expenditures into 2027. About C4 Therapeutics C4 Therapeutics (C4T) (Nasdaq: CCCC) is a clinical-stage biopharmaceutical company dedicated to delivering on the promise of targeted protein degradation science to create a new generation of medicines that transforms patients’ lives. C4T is progressing targeted oncology programs through clinical studies and leveraging its TORPEDO® platform to efficiently design and optimize small-molecule medicines to address difficult-to-treat diseases. C4T’s degrader medicines are designed to harness the body’s natural protein recycling system to rapidly degrade disease-causing proteins, offering the potential to overcome drug resistance, drug undruggable targets and improve patient outcomes. For more information, please visit www.c4therapeutics.com. About CemsidomideCemsidomide is an orally bioavailable MonoDAC® degrader designed to be highly potent and selective against its intended targets of Ikaros (IKZF1) and Aiolos (IKZF3) and overcome shortcomings of currently approved therapies to treat multiple myeloma (MM) and non-Hodgkin’s lymphoma (NHL). Cemsidomide is currently in a Phase 1 dose escalation study in MM and NHL. Initial clinical data show cemsidomide is well tolerated, demonstrates anti-myeloma activity and displays evidence of immunomodulatory effects. More information about this trial may be accessed at www.clinicaltrials.gov (identifier: NCT04756726). About CFT1946CFT1946 is an orally bioavailable BiDAC™ degrader designed to be potent and selective against BRAF V600X mutant targets. In preclinical studies, CFT1946 is active in vivo and in vitro in models with BRAF V600E driven disease and in models resistant to BRAF inhibitors. CFT1946 is currently in a Phase 1 dose escalation study in BRAF V600X mutant solid tumors including colorectal cancer, melanoma and non-small cell lung cancer. More information about this trial may be accessed at www.clinicaltrials.gov (identifier: NCT05668585). Forward-Looking Statements This press release contains “forward-looking statements” of C4 Therapeutics, Inc. within the meaning of the Private Securities Litigation Reform Act of 1995. These forward-looking statements may include, but may not be limited to, express or implied statements regarding our ability to develop potential therapies for patients; the design and potential efficacy of our therapeutic approaches; the anticipated timing and content of presentations of data from our clinical trials; and our ability to fund our future operations. Any forward-looking statements in this press release are based on management’s current expectations and beliefs of future events and are subject to a number of risks and uncertainties that could cause actual results to differ materially and adversely from those set forth in or implied by such forward-looking statements. These risks and uncertainties include, but are not limited to: uncertainties related to the initiation, timing, advancement and conduct of preclinical and clinical studies and other development requirements for our product candidates; the risk that any one or more of our product candidates will cost more to develop or may not be successfully developed and commercialized; and the risk that sufficient capital to fund our future operations will be available to us on acceptable terms or at the times required. For a discussion of these and other risks and uncertainties, and other important factors, any of which could cause our actual results to differ from those contained in the forward-looking statements, see the section entitled “Risk Factors” in C4 Therapeutics’ most recent Annual Report on Form 10-K and/or Quarterly Report on Form 10-Q, as filed with the Securities and Exchange Commission. All information in this press release is as of the date of the release, and C4 Therapeutics undertakes no duty to update this information unless required by law. Contacts: Investors:Courtney SolbergSenior Manager, Investor RelationsCSolberg@c4therapeutics.com Media:Loraine SpreenSenior Director, Corporate Communications & Patient AdvocacyLSpreen@c4therapeutics.com Condensed Consolidated Balance Sheet Data(in thousands) (unaudited) (audited) September 30, 2024 December 31, 2023Cash, cash equivalents and marketable securities$284,400 $281,689Total assets 376,060 376,451Deferred revenue 49,417 37,285Total stockholders' equity 242,656 246,114 Condensed Consolidated Statements of Operations(in thousands, except share and per share amounts)(Unaudited) Three Months Ended September 30, Nine Months Ended September 30, 2024 2023 2024 2023 Revenue from collaboration agreements$15,362 $11,072 $30,407 $17,495 Operating expenses: Research and development 31,838 28,347 78,124 87,315 General and administrative 11,768 10,533 31,751 31,784 Restructuring — — 2,437 — Total operating expenses 43,606 38,880 112,312 119,099 Loss from operations (28,244) (27,808) (81,905) (101,604)Other income (expense), net: Interest expense and amortization of long-term debt − related party — (167) — (1,373)Loss on early extinguishment of debt — (621) — (621)Interest and other income, net 3,578 2,562 11,162 6,862 Total other income, net 3,578 1,774 11,162 4,868 Loss before income taxes (24,666) (26,034) (70,743) (96,736)Income tax expense — (1,003) — (1,003)Net loss$(24,666) $(27,037) $(70,743) $(97,739)Net loss per share − basic and diluted$(0.35) $(0.55) $(1.03) $(1.99)Weighted-average shares outstanding − basic and diluted 69,627,190 49,212,126 68,958,938 49,103,351