作者: Diaz, Olivier ; Perrin-Cocon, Laure ; Lalande, Alexandre ; Jacquemin, Clémence ; Aublin-Gex, Anne ; Mathieu, Cyrille ; Toesca, Johan ; El Orch, Walid ; Jacolin, Florentine ; Si-Tahar, Mustapha ; Vidalain, Pierre-Olivier ; Lotteau, Vincent

AbstractThe hypoxia response pathway enables adaptation to oxygen deprivation. It is mediated by hypoxia-inducible factors (HIF), which promote metabolic reprogramming, erythropoiesis, angiogenesis and tissue remodeling. This led to the successful development of HIF-inducing drugs for treating anemia and some of these molecules are now in clinic. However, elevated levels of HIFs are frequently associated with tumor growth, poor prognosis, and drug resistance in various cancers, including hepatocellular carcinoma (HCC). Consequently, there are concerns regarding the recommendation of HIF-inducing drugs in certain clinical situations. Here, we analyzed the effects of two HIF-inducing drugs, Molidustat and Roxadustat, in the well-characterized HCC cell line Huh7. These drugs increased HIF-1α and HIF-2α protein levels which both participate in inducing hypoxia response genes such as BNIP3, SERPINE1, LDHA or EPO. Combined transcriptomics, proteomics and metabolomics showed that Molidustat increased the expression of glycolytic enzymes, while the mitochondrial network was fragmented and cellular respiration decreased. This metabolic remodeling was associated with a reduced proliferation and a lower demand for pyrimidine supply, but an increased ability of cells to convert pyruvate to lactate. This was accompanied by a higher resistance to the inhibition of mitochondrial respiration by antimycin A, a phenotype confirmed in Roxadustat-treated Huh7 cells and Molidustat-treated hepatoblastoma cells (Huh6 and HepG2). Overall, this study shows that HIF-inducing drugs increase the metabolic resilience of liver cancer cells to metabolic stressors, arguing for careful monitoring of patients treated with HIF-inducing drugs, especially when they are at risk of liver cancer.

2024-11-01·Brain Research Bulletin

Orexin A protects against cerebral ischemia-reperfusion injury by enhancing reperfusion in ischemic cortex via HIF-1α-ET-1/eNOS pathway

Article

作者: Zhu, Yifan ; Guo, Xu ; Zhang, Zhaojun ; Lin, Junwei ; Li, Xiaofeng ; Guo, Jing ; Li, Zhuoqi ; Jiang, Zixuan ; Zhu, Minxia ; Li, Pengfei

The purpose of this study was to investigate the protective effect and underlying mechanism of orexin A on cerebral ischemia-reperfusion injury, specifically through vasodilation mediated by the hypoxia inducible factor-1α (HIF-1α)-Endothelin-1(ET-1)/endothelial nitric oxide synthase (eNOS) pathway. A model of middle cerebral artery occlusion was established in both wild-type SD rats with exogenous orexin A intervention and in orexin A transgenic rats. Neurological deficit scores and cerebral infarction areas were assessed, and ischemic cortical blood flow was monitored. Gene and protein expression levels of HIF-1α, HIF-2α, ET-1, and three types of NOS were detected using real-time RT-qPCR and Western blot analysis, respectively. Additionally, nitric oxide (NO) levels in the cortex were analyzed through biochemical detection methods. Orexin A demonstrated a protective effect by reducing cerebral infarction and improving neurological deficits, which was achieved by increasing cortical blood flow during reperfusion. This protective mechanism was associated with upregulated HIF-1α expression, downregulated ET-1 expression, upregulated eNOS expression, and increased NO production. This study demonstrates the protective effect of orexin A on cerebral ischemia-reperfusion injury, achieved by regulating the release of vasomotor substances to enhance cortical blood flow during reperfusion. These findings suggest that orexin A may represent a potential therapeutic strategy for ischemic stroke.

2024-11-01·Experimental Eye Research

Increased susceptibility of human limbal aniridia fibroblasts to oxidative stress

Article

作者: Stachon, Tanja ; Amini, Maryam ; Zimmermann, Julia Sarah Alexandra ; Chai, Ning ; Seitz, Berthold ; Szentmáry, Nóra ; Fries, Fabian Norbert ; Suiwal, Shweta ; Li, Zhen ; Trusen, Simon

Aniridia-associated keratopathy originates from a haploinsufficiency of the transcription factor PAX6 (PAX6^+/-). In the corneal epithelium of PAX6^+/- mice, a significant increase in oxidized proteins was observed, accompanied by impaired compensation for elevated oxidative stress (OS). The extent to which limbal fibroblast cells (LFCs) are affected by an increased susceptibility to OS in cases of congenital aniridia (AN) has not been determined, yet. Our aim was to examine the impact of OS on antioxidant enzyme expression in normal and AN-LFCs. Following isolation and culture of primary LFCs (n = 8) and AN-LFCs (n = 8), cells were treated with cobalt chloride for 48 h to chemically induce hypoxic conditions and OS. Subsequently, HIF-1α/-2α, PHD1/2, Nrf2, CAT, SOD1, PRDX6, and GPX1 gene expression was examined by qPCR. SOD1, PRDX6, and GPX1 protein levels were assessed from the cell lysate by Western blot. The induction of hypoxia led to reduced HIF-1α gene expression in both fibroblast groups (p≤0.008), while the decrease in PHD1 was limited to AN-LFCs (p = 0.0007). On the other hand, under hypoxic conditions, PHD2 showed higher mRNA expression in AN-LFCs compared to normal LFCs (p = 0.013). As a result of OS, the mRNA levels of Nrf2 (p<0.0001) and the antioxidant enzymes CAT (p = 0.005), SOD1 (p = 0.005), GPX1 (p = 0.002) decreased in AN-LFCs. This was accompanied by an increased protein expression of SOD1 (p = 0.019) and PRDX6 (p=0.0009). In the normal LFC group, the induced extent of OS had no impact on the gene (p≥0.151) and protein expression (p ≥ 0.629) of antioxidant enzymes, except for the GPX1 mRNA level (p = 0.027). AN-LFCs exhibit higher susceptibility to OS than normal LFCs. Therefore, in AN-LFCs, there are sustained alterations in gene and protein expression of antioxidative enzymes even after 48 h of CoCl₂ treatment.

项与 HIF-2α x HIF-1α 相关的新闻（医药）

2024-03-26

·PRNewswire

Research published in Journal of the American Chemical Society highlights potential of HIF Inhibition as a therapeutic approach for cancers

Research led by Curve CSO Professor Ali Tavassoli's laboratory at University of Southampton Landmark paper on dual HIF-1 and HIF-2 inhibitor that works by inhibiting the interaction of both HIF-1α and HIF-2α with HIF-1β SOUTHAMPTON, England, March 26, 2024 /PRNewswire/ -- Curve Therapeutics ("Curve" or the "Company"), a private biotechnology company pioneering a revolutionary intracellular screening platform addressing complex and challenging disease targets, today announces the publication of an article in the Journal of the American Chemical Society (JACS). The paper entitled 'Identification and Development of Cyclic Peptide Inhibitors of Hypoxia Inducible Factors (HIF) 1 and 2 That Disrupt Hypoxia-Related Signalling in Cancer Cells' can be viewed here.1 Curve's Chief Scientific Officer, Professor Ali Tavassoli, co-authored the article and leads the academic group at the University of Southampton where the work was undertaken. Professor Ali Tavassoli, Chief Scientific Officer of Curve Therapeutics, said: "It is well recognised that HIF plays a key role in the survival and growth of solid tumours. The compounds we report in this paper inhibit the protein-protein interaction of the two subunits that form the HIF transcription factor. We show that these compounds prevent the hypoxia-induced activity of this transcription factor, stopping hypoxia-response in cell-based assays. This paper underlines the promising therapeutic potential of dual HIF inhibition as an approach for the treatment of a variety of cancers." Curve Therapeutics is a leader in the discovery of innovative therapeutics to address disease targets which are difficult to target using conventional drug discovery methods. Through the utilisation of its world leading Microcyle® discovery platform, Curve can screen directly inside mammalian cells, allowing for the identification of biologically active library members within an intracellular environment where both the library and the target are present in their native conformations. Curve is developing a non-peptidic, small-molecule dual inhibitor of HIF-1 and HIF-2 with first-in-class potential. This is the first report of a dual HIF-1 and HIF-2 inhibitor which functions by inhibiting the interaction of both HIF-1α and HIF-2α with HIF-1β. The Microcycles discovered by Prof. Tavassoli were identified using his SICLOPPS screening platform and show good cellular activity. Patents and patent applications describing these HIF inhibitory Microcycles are exclusively licensed to Curve. About Curve Therapeutics Curve Therapeutics is a private biotechnology company pioneering a revolutionary intracellular screening platform to enable the discovery of innovative therapeutics that address complex and challenging disease targets with the potential to transform the lives of patients. Curve originated from world-leading Microcycle® research conducted by Professor Tavassoli's group in the Department of Chemistry at the University of Southampton, UK. Curve is backed by blue chip investors including Advent Life Sciences, Epidarex Capital, Pfizer Ventures, Columbus Venture Partners and British Patient Capital. Curve has a US$1.7bn global research collaboration with MSD the trade name of Merck & Co., Inc., Kenilworth, NJ USA, to discover and validate modulators of up to five therapeutic targets using its Microcycle® technology, initially for oncology and neurology indications. For more information visit: . About Curve's Microcycle® platform Curve has developed an IP-protected, mammalian cell platform technology for functional screening and enrichment of diverse hexameric cyclic peptide Microcycle® libraries to identify those library members that have the desired biological activity against a therapeutic target. Curve's platform allows direct screening for biologically active library members inside mammalian cells and facilitates small molecule hit-to-lead programmes. A key advantage of the technology is that both the library and the target are present in all of their native conformations within a cell. Uniquely, the compact size and rigid structure of Microcycles® enables the design of non-peptide small molecule leads. The platform can be used for a wide range of therapeutically relevant targets, including protein-protein and protein-DNA interactions and has been used by Curve to develop a pipeline of cancer programmes against targets including a dual HIF-1/HIF-2 inhibitor and an inhibitor of ATIC homodimerization. [1] Andrew T. Ball, Soran Mohammed, Cyrielle Doigneaux, Reece M. Gardner, James W. Easton, Steven Turner, Jonathan W. Essex, Garry Pairaudeau, Ali Tavassoli, Identification and Development of Cyclic Peptide Inhibitors of Hypoxia Inducible Factors 1 and 2 That Disrupt Hypoxia-Response Signaling in Cancer Cells, Journal of the American Chemical Society (19 March 2024) DOI: 10.1021/jacs.3c10508 SOURCE Curve Therapeutics

2023-12-12

·今日头条

双靶向抑制剂！沈阳药科大学陈国良/王立辉团队发现癌症发展最新分子机制

本文为转化医学网原创，转载请注明出处作者：Rainbow 导读：肺癌是全球最常见的癌症之一，也是导致癌症相关死亡的主要原因，每年估计有200万新病例和176万人死亡。 12月10日，沈阳药科大学陈国良、王立辉共同通讯在期刊《ADVANCED SCIENCE》上在线发表题为“The Interplay Between HIF-1α and EZH2 in Lung Cancer and Dual-Targeted Drug Therapy”的研究论文，研究证明了表观遗传修饰酶EZH2参与了HIF-1α抑制剂耐药过程。抑制HIF-1α会导致PRC2复合物中SUZ12的转录抑制丧失，进而增加EZH2酶活性。同时，EZH2作为HIF-1α的转录抑制因子。HIF-1α和EZH2相互作用形成负向反馈环路，促进癌症发展。双靶向抑制剂DYB-03在体外和体内结合EZH2和HIF-1α，发挥抗肿瘤作用。结果为双靶向HIF-1α和EZH2提供了药理学基础。 https://onlinelibrary.wiley.com/doi/10.1002/advs.202303904 研究背景 01 肺癌是全球最常见的癌症之一，也是导致癌症相关死亡的主要原因，每年估计有200万新病例和176万人死亡。非小细胞肺癌（NSCLC）根据组织病理类型的诊断约占所有病例的85%。NSCLC的治疗异常复杂，原因在于存在同时发生的基因组变异、代谢重编程和表观遗传异常。低氧是NSCLC等实体肿瘤的一个显著特征。肿瘤内低氧和反应性氧化物种、细胞因子和生长因子等信号分子的参与导致HIF（低氧诱导因子）高度活化。HIF是一个转录因子，由两个亚基组成，即HIF-1α（或其类似物HIF-2α和HIF-3α）和HIF-1β。表观遗传修饰是一种生物学过程，其中DNA序列保持不变，但由于可逆的调节作用，主要包括DNA甲基化和组蛋白修饰，表型被改变。表观遗传失调在肿瘤的发展、进展和获得治疗抵抗中发挥着重要作用，包括实体肿瘤和血液系统恶性肿瘤。除了在单药治疗中潜在的作用外，表观遗传药物可能在与其他抗癌疗法协同作用或逆转耐药方面发挥重要作用。 DYB-03通过靶向 HIF-1和EZH2发挥抗肿瘤功能 02 研究人员建立了一个稳定的EZH2/HIF-1α双基因敲除细胞系。与亲本细胞相比，EZH2/HIF-1α双基因敲除细胞在增殖和迁移能力上部分丧失，表明这两种蛋白在肿瘤发生中起到了作用。集落形成和划痕愈合实验显示，与对照相比，DYB-03对双基因敲除细胞的增殖和迁移没有显著影响。Transwell实验显示，DYB-03抑制细胞迁移的能力消失，但当重新输入HIF-1α和EZH2时，这种抑制效果被逆转，表明DYB-03作用于HIF-1和EZH2。研究人员在体内验证了DYB-03的靶向特异性，并显示双基因敲除HIF-1α和EZH2可以减缓A549的肿瘤生长，类似于DYB-03对亲本细胞的抑制效果。此外，DYB-03的给药对A549双基因敲除细胞的肿瘤生长没有显著抑制作用，表明DYB-03通过作用于HIF-1α和EZH2发挥其抗肿瘤活性。免疫印迹验证了A549双基因敲除细胞的蛋白稳定性。肿瘤组织的组织学分析还检测了Ki67和CD31的表达。双基因敲除肿瘤组织中Ki67和CD31的表达要比亲本肿瘤组织低得多。DYB-03的给药在亲本组中显示出明显的抗增殖和抗血管生成作用，而在A549双基因敲除组中作用较小。上述结果表明，DYB-03通过作用于HIF-1和EZH2在体内也发挥其抑制肿瘤的效果。研究结果 03 综上所述，研究证明了表观遗传修饰酶EZH2参与了HIF-1α抑制剂耐药过程。抑制HIF-1α会导致PRC2复合物中SUZ12的转录抑制丧失，进而增加EZH2酶活性。同时，EZH2作为HIF-1α的转录抑制因子。HIF-1α和EZH2相互作用形成负向反馈环路，促进癌症发展。双靶向抑制剂DYB-03在体外和体内结合EZH2和HIF-1α，发挥抗肿瘤作用。结果为双靶向HIF-1α和EZH2提供了药理学基础。参考资料： https://onlinelibrary.wiley.com/doi/10.1002/advs.202303904 注：本文旨在介绍医学研究进展，不能作为治疗方案参考。如需获得健康指导，请至正规医院就诊。热门·直播/活动 🕓 北京｜12月19日-20日 ▶第四届单细胞测序技术应用研讨会暨单细胞&空间组学研讨会（日程稍后公布）点击对应文字查看详情

临床结果临床研究

2023-08-20

·药研发

【药研发0821】尚健抗肿瘤1类新药获批临床 | 2023年国家医保目录调整结果公布...

「本文共：15条资讯，阅读时长约：3分钟」今‍日‍头条尚健抗肿瘤1类新药获批临床。尚健生物1类生物制品注射用SG1827获国家药监局临床试验默示许可，拟开发用于治疗晚期恶性实体瘤。SG1827可以在TCR刺激信号存在的情况下，特异性介导共刺激信号，激活T淋巴细胞。临床前研究显示，SG1827单药在多种肿瘤模型中显示出积极抗肿瘤活性；与PD-1抗体等免疫检查点抑制剂联用，协同发挥抗肿瘤活性。‍‍‍国‍‍‍‍‍‍‍‍‍内‍‍‍‍药‍‍讯‍‍‍‍1.强生BCMA/CD3双抗中国报产。强生旗下杨森BCMA/CD3双特异性抗体特立妥单抗注射液（teclistamab）的上市申请获CDE受理，用于治疗复发或难治性多发性骨髓瘤（r/rMM）。特立妥单抗旨在将CD3阳性T细胞募集到表达BCMA的骨髓瘤细胞附近，并且激活T细胞对肿瘤细胞进行攻击。去年8月，该新药首次获欧盟批准上市，用于治疗接受至少三种前期疗法的r/rMM患者，成为全球首款BCMA/CD3双抗。2.天境长效生长激素III期临床成功。天境生物在2023年H1财报中透露，该公司引进的新型伊坦生长激素（TJ101,eftansomatropin alfa）在治疗儿童生长激素缺乏症的III期研究达到主要终点。与Norditropin（诺泽）相比，TJ101组第52周患者年化身高生长速率（AHV）达到非劣效性（10.76cm/年vs10.28cm/年，P<0.0001)。天境生物计划明年提交伊坦生长激素的上市申请。3.和黄MET抑制剂拟纳入胃癌突破性品种。和黄医药口服小分子MET-TKI沃利替尼片（savolitinib）获CDE拟纳入突破性治疗品种，用于治疗既往至少二线标准治疗失败的MET基因扩增的局部晚期或转移性胃癌或胃食道连接部腺癌。savolitinib可阻断因突变(例如外显子14跳跃突变或其它点突变)、基因扩增或蛋白质过表达而导致的MET受体酪氨酸激酶信号通路的异常激活。此前，该新药已获批用于治疗MET外显子14跳跃突变的非小细胞肺癌患者。4.石药CLDN6/CD137双抗获批实体瘤临床。石药集团旗下公司Novarock开发的CLDN6/CD137双抗NBL-028获FDA批准，即将开展用于治疗CLDN6表达晚期肿瘤患者的临床试验。NBL-028在与TME中的肿瘤细胞表面CLDN6结合的同时，选择性地激活T细胞及其他免疫细胞中的CD137共激通路，从而达到选择性杀伤肿瘤细胞的作用。该新药有望为睾丸癌、卵巢癌、非小细胞肺癌及子宫内膜癌等肿瘤患者提供新的治疗选择。5.恒瑞上半年创新药营收50亿元。恒瑞医药公布2023年上半年业绩报告，上半年营收111.68亿元，同比增长9.19%；归属于上市公司股东的净利润23.08亿元，同比增长8.91%；研发投入23.31亿元，同比增长6.73%。目前，恒瑞医药已有13款自研创新药、2款引进创新药在国内获批上市，报告期内公司创新药收入达到49.62亿元（含税）。‍‍‍‍国‍‍‍‍‍‍‍‍‍‍际‍‍‍‍药‍讯‍‍1.再生元新一代C5单抗获批上市。再生元补体因子C5靶向单抗Veopoz（pozelimab）获FDA批准上市，成为针对CD55缺陷型蛋白丢失性肠病（CHAPLE）的首款疗法。这是一种由编码CD55蛋白的基因突变而无法调节补体活动导致的罕见遗传免疫疾病。在Ⅱ/Ⅲ期临床中，24周治疗结果显示，100%患者的血清白蛋白快速且持续恢复正常，且临床症状改善或无恶化。2.VMAT2抑制剂获批治疗亨廷顿式病。Neurocrine公司VMAT2抑制剂Ingrezza（缬苯那嗪,Valbenazine）获FDA批准新适应症，用于治疗亨廷顿式病成人患者的舞蹈症状。III期研究结果显示，与安慰剂相比，Valbenazine治疗组患者16周时疾病评定量表（UHDRS）最大舞蹈症（TMC）评分显著降低（-4.6 vs. -1.4分；P<0.0001）。2017年4月，FDA已批准INGREZZA上市，用于治疗迟发性运动障碍（tardive dyskinesia）。3.再生元CD3/CD20双抗向EMA报MAA。欧洲药品管理局（EMA）受理再生元CD3/CD20双抗odronextamab（REGN1979）上市许可申请（MAA），用于治疗复发/难治性滤泡性淋巴瘤（FL）和弥漫性大B细胞淋巴瘤（DLBCL）。在II期试验中，随访为17.1个月时，该新药在DLBCL患者中ORR和CR分别为53%和37%；在FL患者中，odronextamab的ORR和CR分别为82%和75%。再鼎医药拥有该产品的中国开发和商业化权益。4.默沙东HIF-2α抑制剂肾癌Ⅲ期临床积极。默沙东“first-in-class” 缺氧诱导因子-2α（HIF-2α）抑制剂Welireg，在治疗经治晚期肾细胞癌的Ⅲ期临床LITESPARK-005达到主要终点。与活性对照相比，Welireg显著改善患者的无进展生存期（PFS）；总生存期（OS）也呈改善趋势，但未达到统计学显著性。2021年8月，FDA已加速批准Welireg上市，成为首个HIF-2α抑制剂，用于治疗包括肾细胞癌（RCC）等von Hippel-Lindau（VHL）疾病相关癌症。5.UGT1A1靶向基因疗法肝病早期临床积极。Généthon实验室靶向UGT1A1的AAV8基因疗法GNT0003治疗Crigler-Najjar综合征的Ⅰ/Ⅱ期临床结果发表在《新英格兰医学杂志》上。Généthon旨在通过一次性治疗，恢复患者的肝脏细胞生成UGT1A1的功能，降低胆红素的水平。试验结果显示，最高剂量组的3例患者的血清胆红素水平显著下降，且疗效可维持长达80周。此外，临床中没有发现严重副作用。6.神经疾病口服肠菌疗法获罕见儿科疾病认定。Bloom Science公司口服肠道微生物治疗产品BL-001获FDA授予罕见儿科疾病认定，用于治疗Dravet综合征神经疾病。BL-001旨在复制生酮饮食的抗癫痫作用，调节γ-氨基丁酸（GABA）和其他关键的生物能量途径。在Ⅰ期临床中，BL-001在所有4个剂量组中均表现出良好的安全性和耐受性，支持比动物模型中的有效剂量水平高出10倍的用药剂量。‍‍‍‍‍‍‍‍‍医‍‍‍‍‍‍药‍‍‍‍‍‍‍‍‍‍‍‍‍热‍‍‍‍‍‍‍点‍‍‍。1.2023年国家医保目录调整结果公布。8月18日，国家医保局发布《关于公示2023年国家基本医疗保险、工伤保险和生育保险药品目录调整通过初步形式审查的药品及相关信息的公告》，公示时间为：2023年8月18日-8月24日（一周）。此轮参与医保目录调整的申报信息共计629份，涉及药品570个，申报药品数目较去年有一定增加。经初步审查，390个药品通过初步形式审查。通过形式审查不等于纳入国家医保药品目录，后续还有专家评审、谈判竞价，预计11月公布新版目录。2.湖南省乡村医生等级评定试点方案出台。近日，湖南省卫健委印发《湖南省乡村医生等级评定试点工作方案》，旨在通过分级评定、分类保障，绩效考核、安排入编等指标来加强基层人才队伍建设，保障乡村医生合理收入。根据《方案》补贴政策：一级村医每月补贴不低于800元；二级村医不低于500元；三级村医补贴标准是不低于300元。3.中南大学湘雅医院揭示二甲双胍新药效。近日，中南大学湘雅医院大型研究团队使用小鼠模型来确定二甲双胍在骨质疏松和正常骨折愈合中的骨再生潜力的研究论文最新发表在《Bone Research》上。研究发现，在骨折愈合过程中，二甲双胍增加了H型血管和骨形成。其机制可能是二甲双胍通过抑制内皮细胞YAP1/TAZ，增加HIF-1α表达，从而促进体内外血管生成。‍‍‍‍‍‍‍评‍‍‍‍‍审‍‍‍‍动‍‍态‍‍‍‍‍‍‍ 1. CDE新药受理情况（08月19日) 2. FDA新药获批情况（北美08月20日）股市资讯上个交易日 A 股医药板块 -2.82%涨幅前三跌幅前三昊海生科 +5.99% 启迪药业-10.03%*ST 太安 +5.04% 赛隆药业-10.01%百奥泰 +3.66% 通化金马 -9.93% 【片仔癀】2023年上半年实现营业收入50.45亿元同比增长14.08%;归属于上市公司股东的净利润15.41亿元，同比增长17.23%;归属于上市公司股东的扣除非经常性损益的净利润15.77亿元，同比增长19.80%。【舒泰坤】STSA-1002皮下注射液(ANCA相关性血管炎适应症)I期临床(FDA)实验结果显示安全性和耐受性良好。【奥赛康】子公司生物创新药ASKG915在美国开展的I期临床研究完成首例患者给药其他- The End -戳“阅读原文”，了解更多医药研发及股市资讯。

财报临床3期突破性疗法申请上市