Ovarian clear cell carcinoma (OCCC) is a unique clinicopathological subtype of epithelial ovarian cancer that is resistant to standard chemotherapy. Eribulin, a microtubule dynamics inhibitor of halichondrin class, has unique effects in the cancer microenvironment such as induction of epithelization and reduction in metastatic potential in breast cancer cells; however, nothing is known about the effect of eribulin and the detailed mechanisms in OCCC. This study aimed to investigate the involvement of ferroptosis and its mechanism in the antitumor activity of eribulin in OCCC cells and a mouse xenograft model. We found that eribulin-induced cell death was reduced by ferroptosis inhibitors; deferoxamine, an iron chelator and ferrostatin-1, a lipid peroxidation inhibitor. Eribulin increased the levels of intracellular iron, reactive oxygen species (ROS), and lipid peroxides, and increased the mitochondrial membrane potential. Eribulin downregulated the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), the mitochondrial enzyme dihydroorotate dehydrogenase (DHODH), and superoxide dismutase (SOD) activity. The combination of eribulin and ML210, a glutathione peroxidase 4-inhibiting ferroptosis inducer, had a synergistic effect on ferroptosis. Taken together, our findings show firstly that eribulin triggers ferroptosis in OCCC and this effect occurs via the suppression of the Nrf2-HO-1 signaling pathway, SOD activity and the promotion of lipid peroxidation. These findings suggest that eribulin-induced ferroptosis is associated with its anti-tumor effect and also could be a potential therapeutic target in OCCC.

2024-02-08·Journal of Medicinal Chemistry1区 · 医学

Novel Covalent Probe Selectively Targeting Glutathione Peroxidase 4 In Vivo: Potential Applications in Pancreatic Cancer Therapy

1区 · 医学

Article

作者: Li, Zhengqiu ; Zhu, Chengjun ; Xu, Fang ; Tan, Yi ; Tang, Zifeng ; Li, Jie ; Sun, Pinghua ; He, Xiaoqiang ; Zhang, Zhang ; Peng, Lijie ; Zhang, Zhi-Min ; Ding, Ke

Glutathione peroxidase 4 (GPX4) emerges as a promising target for the treatment of therapy-resistant cancer through ferroptosis. Thus, there is a broad interest in the development of GPX4 inhibitors. However, a majority of reported GPX4 inhibitors utilize chloroacetamide as a reactive electrophilic warhead, and the selectivity and pharmacokinetic properties still need to be improved. Herein, we developed a compound library based on a novel electrophilic warhead, the sulfonyl ynamide, and executed phenotypic screening against pancreatic cancer cell lines. Notably, one compound A16 exhibiting potent cell toxicity was identified. Further chemical proteomics investigations have demonstrated that A16 specifically targets GPX4 under both in situ and in vivo conditions, inducing ferroptosis. Importantly, A16 exhibited superior selectivity and potency compared to reported GPX4 inhibitors, ML210 and ML162. This provides the structural diversity of tool probes for unraveling the fundamental biology of GPX4 and exploring the therapeutic potential of pancreatic cancer via ferroptosis induction.

2023-04-01·Pharmacology & Therapeutics

Molecular mechanisms of ferroptosis and their involvement in brain diseases

Review

作者: Silva, Vera ; Benfeito, Sofia ; Silva, Renata ; Barbosa, Daniel José ; Borges, Fernanda ; Remião, Fernando ; Chavarria, Daniel ; Costa, Inês

Ferroptosis is a type of regulated cell death characterized by intracellular accumulation of iron and reactive oxygen species, inhibition of system Xc-, glutathione depletion, nicotinamide adenine dinucleotide phosphate oxidation and lipid peroxidation. Since its discovery and characterization in 2012, many efforts have been made to reveal the underlying mechanisms, modulating compounds, and its involvement in disease pathways. Ferroptosis inducers include erastin, sorafenib, sulfasalazine and glutamate, which, by inhibiting system Xc-, prevent the import of cysteine into the cells. RSL3, statins, Ml162 and Ml210 induce ferroptosis by inhibiting glutathione peroxidase 4 (GPX4), which is responsible for preventing the formation of lipid peroxides, and FIN56 and withaferin trigger GPX4 degradation. On the other side, ferroptosis inhibitors include ferrostatin-1, liproxstatin-1, α-tocopherol, zileuton, FSP1, CoQ10 and BH4, which interrupt the lipid peroxidation cascade. Additionally, deferoxamine, deferiprone and N-acetylcysteine, by targeting other cellular pathways, have also been classified as ferroptosis inhibitors. Increased evidence has established the involvement of ferroptosis in distinct brain diseases, including Alzheimer's, Parkinson's and Huntington's diseases, amyotrophic lateral sclerosis, multiple sclerosis, and Friedreich's ataxia. Thus, a deep understanding of how ferroptosis contributes to these diseases, and how it can be modulated, can open a new window of opportunities for novel therapeutic strategies and targets. Other studies have shown a sensitivity of cancer cells with mutated RAS to ferroptosis induction and that chemotherapeutic agents and ferroptosis inducers synergize in tumor treatment. Thus, it is tempting to consider that ferroptosis may arise as a target mechanistic pathway for the treatment of brain tumors. Therefore, this work provides an up-to-date review on the molecular and cellular mechanisms of ferroptosis and their involvement in brain diseases. In addition, information on the main ferroptosis inducers and inhibitors and their molecular targets is also provided.

项与 ML-210 相关的新闻（医药）

2022-10-16

·BiG生物创新社

北大刘国全团队 | 靶向光降解PDTAC引发铁死亡和有效的抗肿瘤免疫

导读铁死亡（ferroptosis）是近年发现的一种新型细胞死亡方式，其特征是谷胱甘肽耗竭，谷胱甘肽过氧化物酶 4 (GPX4 ) 活性下降；脂质氧化物不能被还原，继而二价铁离子氧化脂质产生大量活性氧，导致细胞发生铁死亡。由于该过程依赖铁，所以被称为铁死亡。作为铁死亡核心调控子的GPX4，已被认为是细胞铁死亡研究的“明星分子”用小分子抑制剂靶向GPX4进行癌症治疗仍然具有挑战性。GPX4的分子表面缺乏药物样结合袋。经典抑制剂（例如，RSL3 和 ML210）均通过与GPX4的活性位点共价结合而发挥作用，这在体内应用中可能具有低选择性和全身毒性。光降解靶向嵌合体 (PDTAC)由三个功能模块组成，一个结合感兴趣的蛋白质 (POI) 的靶向配体，一个在光照射下产生活性氧 (ROS) 的光敏剂 (PS)，以及一个连接前两部分的接头。原则上，PDTAC 的三个模块可以组装成通用分子来降解所需的蛋白质。蛋白质降解与光激活的耦合赋予 PDTAC 高度时空控制，这将非常有利于药物开发。2022年9月6日，北京大学药学院化学生物学系，天然药物与仿生药物国家重点实验室课题组长刘国全团队在药物化学权威期刊Journal of Medicinal Chemistry在线发表了题为“PDTAC: Targeted Photodegradation of GPX4 Triggers Ferroptosis and Potent Antitumor Immunity”的最新研究成果。研究者创新性地将维替泊芬(VPF)与GPX4靶向配体结合设计了PDTAC可以选择性地降解 GPX4并诱导具有强免疫原性的铁死亡细胞死亡。下面就跟随小编一起来解读这篇文献吧。作者 | 阿卜图一光降解GPX4和触发铁死亡的原理示意图靶向嵌合体的表征既然是靶向+产生活性氧，作者首先选择了文献报道的在红光激发下具有很强产生单线态氧的光敏剂维替泊芬，以及两个靶向肽TP-1和TP-2合成了两个PDTAC(PV-1和PV-2)。在作者先前研究中就探究了不同的光敏剂通过PDT诱导的铁死亡。拿到目标产物作者马不停蹄的想要去验证其效果。作者检查了这些嵌合体是否可以穿透癌细胞。两个PDTAC在A549细胞中孵育，基于VPF部分荧光和绿色的商用膜染料Dio共聚焦成像表明，这两种嵌合体很容易被A549细胞吸收。紧接着研究这些嵌合体对GPX4的结合亲和力，作者的初衷是光敏剂和靶向肽联合发挥各自的功能但彼此之间不影响。使用BLI测量了亲和常数KD。结果表明单独的光敏剂VPF不与GPX4mu结合，与它们的靶向肽相比，两种嵌合体的KD几乎不受影响。此外，评估了PV-2与GPX4在活细胞中的靶向结合，细胞热位移测定实验（CTSA）表明PV-2确实增加了GPX4的热稳定性。剂量依赖性实验显示在μM尺度上的有效浓度与从BLI获得的KD相当。靶向功能验证完毕，你和作者想到一起去了。为了测试这些嵌合体是否可以光降解GPX4mu，作者使用基于考马斯蓝的Bradford蛋白测定来监测光解过程。照射15分钟后，两种嵌合体在SDS-PAGE分析中几乎消除了GPX4mu条带，而单独的VPF对GPX4mu仅表现出中等影响。这些结果主要证明了作者设计的想法，即靶向肽引导光诱导的ROS有效降解POI。图二 PDTAC分子的表征在细胞裂解物中选择性光解GPX4不要以为完结撒花了，能够发表高水平文章，让我们接着跟随作者的脚步探索科研之旅。作者为了验证这些了两个PDTAC的红光激发下是否可以降解细胞裂解物中的GPX4。当暴露在光照下5分钟时，两种嵌合体都可以显着降解GPX4，并表现出明显的浓度依赖性。此外，当嵌合体浓度固定时，GPX4的降解也取决于辐照持续时间。GPX4水平在没有光照射的情况下根本不受影响，这表明降解是光诱导ROS的结果。有趣的是，这种降解具有高度选择性，而GPX家族的另一个成员谷胱甘肽过氧化物酶1(GPX1)的丰度保持不受影响。此外，其他蛋白质，例如酰基辅酶A合成酶长链家族成员4（ACSL4，铁死亡的重要调节因子）和乙酰辅酶A乙酰转移酶1（ACAT1，调节甾醇代谢），在相同处理下也没有受到影响。图三PV-1和PV-2在细胞裂解物中选择性降解GPX4在活细胞中选择性光解GPX4作者一鼓作气，研究了这些嵌合体是否保留了它们在复杂细胞环境中光降解GPX4的能力。GPX4也被PV-1或PV-2照射降解，这取决于照射持续时间和嵌合体浓度。没有光照射的嵌合体不影响GPX4水平再次证实了光诱导ROS在这种活细胞蛋白质降解中的关键作用。检查其他蛋白质的丰度，包括ACSL4、GPX1 和 ACAT1，也证明了PV-1 和PV-2光解的选择性。此外，使用考马斯蓝通过SDS-PAGE对整个细胞蛋白进行染色，以检查光降解的选择性。有趣的是，在PV-2处理的细胞中观察到对应于GPX4的条带中的褪色，而其他条带与未处理细胞中的条带相似。图四 PV-1 和 PV-2 在 A549 活细胞中选择性降解 GPX4GPX4的光解主要诱导铁死亡作者评估了对铁死亡高度敏感的癌细胞中的细胞死亡，例如三阴性乳腺癌（MDA-MB-231和4T1）和黑色素瘤（鼠B16）。MDA-MB-231、4T1和B16细胞的IC50值分别为2.8、3.7和3.8μM。观察到Lip-1 不仅降低了脂质ROS水平，而且还降低了由PDT引起的细胞死亡。另外铁死亡抑制剂，铁螯合剂去铁胺（DFO），也显着降低了由这些嵌合体引发的细胞死亡。相比之下，坏死性细胞死亡通常是高PS浓度下的主要细胞死亡途径。如台盼蓝染色所示，具有4-5μM VPF的 PDT 导致细胞膜快速破坏，而用嵌合体处理的细胞仍然完好无损。Lip-1和 DFO都不能降低由 VPF 引起的细胞死亡。图五带有PV-2 的PDT 在多个癌细胞中诱导铁死亡GPX4的光解引发有效的免疫原性细胞死亡在细胞层面研究了嵌合体的光降解效果，作者紧接着通过用PV-2处理的垂死B16细胞接种免疫能力强的小鼠，观察到延长的存活率和当小鼠再次用活癌细胞接种时延缓肿瘤生长。带有一些非靶向PS的PDT也可能具有免疫原性。小鼠骨髓来源的树突状细胞(BMDCs)的体外刺激表明，PV-2处理的癌细胞比VPF处理的细胞导致更有效的BMDC成熟，这一点从CD11c+中 MHC II和CD86 的表达水平所可以看出来。对垂死的癌细胞释放的ATP和高迁移率组蛋白B1（HMGB1）的测量也表明，PV-2处理诱发了这两种常见的DAMPs的更高浓度。这种通过PDTAC对蛋白质进行的选择性光降解具有快速出现的 PROTAC（蛋白水解靶向嵌合体）技术的一些特征。GPX4的光解表现出至少两个有意义的特征，即使用非抑制性靶向配体，尤其是降解难成药物蛋白靶标。此外，PDTAC拥有光激活的高时空精度，这有助于减少体内使用时的全身毒性。另外，PS的多种选择可以导致使用近红外光进行光解，这是体内应用的最佳条件。图六带有PV-2的PDT在体外和体内均诱导ICD总结作者提出了一种独特的PDTAC策略，该策略由三个可替换模块组成，以诱导蛋白质的光降解。为了证明其可行性，通过将不具有抑制活性的肽配体与PS合成了两种靶向嵌合体，旨在基于GPX4的靶向降解引发癌细胞中的铁死亡。这些嵌合体保留了对GPX4的结合亲和力，并在红光照射下选择性地降解细胞裂解物和活细胞中的GPX4。这种酶的光降解诱导脂质过氧化，并主要将细胞死亡转变为铁死亡，从而在体外和体内触发有效的免疫原性反应。基于PDTAC策略的GPX4靶向降解为诱导铁死亡提供了一种新的光可实现的方法，这可能会促进其在癌症免疫治疗中的应用。主要参考文献 [1] Liu S, Zhao X, Shui S, Wang B, Cui Y, Dong S, Yuwen T, Liu G. PDTAC: Targeted Photodegradation of GPX4 Triggers Ferroptosis and Potent Antitumor Immunity. J Med Chem. 2022 Sep 22;65(18):12176-12187. doi: 10.1021/acs.jmedchem.2c00855. Epub 2022 Sep 6. PMID: 36066386.[2] Shui S, Zhao Z, Wang H, Conrad M, Liu G. Non-enzymatic lipid peroxidation initiated by photodynamic therapy drives a distinct ferroptosis-like cell death pathway. Redox Biol. 2021 Sep;45:102056. doi: 10.1016/j.redox.2021.102056. Epub 2021 Jun 23. PMID: 34229160; PMCID: PMC8264218.[3] van Straten D, Mashayekhi V, de Bruijn HS, Oliveira S, Robinson DJ. Oncologic Photodynamic Therapy: Basic Principles, Current Clinical Status and Future Directions. Cancers (Basel). 2017 Feb 18;9(2):19. doi: 10.3390/cancers9020019. PMID: 28218708; PMCID: PMC5332942.[4] Sakamoto K, Sogabe S, Kamada Y, Matsumoto SI, Kadotani A, Sakamoto JI, Tani A. Discovery of GPX4 inhibitory peptides from random peptide T7 phage display and subsequent structural analysis. Biochem Biophys Res Commun. 2017 Jan 8;482(2):195-201. doi: 10.1016/j.bbrc.2016.11.035. Epub 2016 Nov 9. PMID: 27836545.扫码加入BiG生物创新社读者交流群，分享、交流纯粹的行业知识，非诚勿扰！BiGScientific Driven, Making Impact!创新生态丨医药论坛丨行业分析媒体公关丨BiG Webinar联系我们商务：Max 18662346610媒体：Kathy 17621909690

蛋白降解靶向嵌合体免疫疗法小分子药物

100 项与 ML-210 相关的药物交易

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