Erastin   中文名称: 爱拉斯汀

VDAC2; VDAC3

异位子宫内膜基质细胞 (EESC) 中的铁死亡由 erematin（10 μM；24 小时）触发，9 小时后，总 ROS 水平上升 [1]。在 EESC 细胞中，红十字蛋白可以缩短线粒体长度并提高其膜密度 [1]。当用红菊酯 (10 μM) 处理 9 小时时，EESC 中铁相关蛋白（包括 FPN（铁输出蛋白））的 mRNA 表达水平较低。另一方面，FPN 的过度表达可以显着预防 Erastin 诱导的 EESC 铁死亡[1]。在 HT-29 结直肠癌细胞中，erematin（10 μM；24 小时）会导致线粒体通透性转换孔 (mPTP) 打开 [2]。 eratin（30 μM；72 小时）可显着抑制 HT-29 结直肠癌细胞的增殖 [2]。控制铁代谢或线粒体脂肪酸代谢的基因参与促红细胞生成素引发铁死亡的生物学机制。包含四肽重复结构域 35、柠檬酸合酶、ATP 合酶 F0 复合体亚基 C3、核糖体蛋白 L8、铁反应元件结合蛋白 2 (IREB2) 和酰基辅酶 A 合成酶家族成员 2 (ACSF2)[3]。

可以用Erastin创建铁死亡诱导的动物模型。在子宫内膜异位症小鼠模型中，Erastin（40 mg/kg；腹腔注射；每 3 天一次，持续 2 周）抑制子宫内膜异位症着床，表明 Erastin 通过诱导铁死亡促进异位病变消退 [1]。在 SCID 小鼠中，eratin（10 mg/kg、30 mg/kg；腹腔注射；每天一次，持续 4 周）抑制 HT-29 异种移植物的生长，其中 30 mg/kg 显示出最大活性 [2]。

Erastin抑制电压依赖性阴离子通道（VDAC2/VDAC3）并加速氧化，导致内源性活性氧的积累。
我们在此评估了erastin（一种电压依赖性阴离子通道（VDAC）结合化合物）潜在的抗结肠癌症活性。我们的体外研究表明，erastin可能通过诱导氧化应激和胱天蛋白酶-9依赖性细胞凋亡，对多种人类结直肠癌癌症细胞系发挥了强大的细胞毒性作用。此外，在erastin处理的癌症细胞中观察到线粒体通透性转变孔（mPTP）开放，这通过VDAC-1和亲环蛋白-D（Cyp-D）结合、线粒体去极化和细胞色素C释放证明。胱天蛋白酶抑制剂、ROS清除剂MnTBAP和mPTP阻断剂（桑格列非林A、环孢菌素A和邦克雷酸），以及shRNA介导的VDAC-1敲低，都显著减弱了erastin诱导的结直肠癌癌症细胞的细胞毒性和凋亡。另一方面，VDAC-1的过度表达增加了erastin诱导的ROS产生、mPTP开放和结直肠癌癌症细胞凋亡。体内研究表明，在严重联合免疫缺陷（SCID）小鼠中，以耐受性良好的剂量腹腔注射erastin可显著抑制HT-29异种移植物的生长。总之，这些结果表明erastin对癌症细胞具有细胞毒性和促凋亡作用。Erastin可以作为一种新型的抗癌症药物被进一步研究。

细胞活力测定[1]
细胞类型：正常子宫内膜基质细胞 (NESC) 和子宫内膜基质细胞 (EESC)
测试浓度： 0、0.5、 0.8、1、1.5、2、2.5、5、10 μM
孵育时间： 24 小时
实验结果：诱导细胞脱离和明显EESC 死亡，但 NESC 不死亡。

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细胞凋亡分析[1]
细胞类型：感染表达 FPN cDNA 的腺病毒的 EESC（共孵育 24 小时）
测试浓度： 0、0.5、1.5、2.5、5 和 2.5 μM
孵育时间： 24 小时
实验结果：通过降低总 ROS 和脂质 ROS 水平。并通过腺病毒感染细胞中 FPN 的过度表达而逆转。

Animal/Disease Models: Mouse model of endometriosis[1]
Doses: 40 mg/kg
Route of Administration: intraperitoneal (ip)injection; once every 3 days for 2 weeks
Experimental Results: demonstrated little impact on body weight of mice and hair of mice displayed neat and glossy. decreased the volume of ectopic lesions.

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Mouse model of endometriosis[1]
Ten C57BL/6 female mice (7–8 weeks, weight 20–22 g) were used. Endometriotic lesions were surgically induced by autotransplantation of uterine horns onto the peritoneal wall as previously described. Briefly, uterine horns were removed and opened longitudinally, cut into homogeneous fragments using a 3-mm dermal biopsy punch and then transplanted onto own peritoneal wall of mice by suturing. 17-β-Estradiol-3-benzoate (30 μg/kg) was administered to each postoperative mouse every 3 days for 28 days. At 14 day after operation, endometrial-like lesions were established, and it was time for intervention. They were randomly divided into two groups. In the experimental group, each mouse received erastin (40 mg/kg) by intraperitoneal injection over a 14-day period. In the control group, in place of erastin, soybean oil was used. At 28 days, the mice were sacrificed and we harvested the ectopic tissues. The volumes of ectopic lesions were measured and analyzed as previously described (Zhao et al., 2015).

[1]. Erastin induces ferroptosis via ferroportin-mediated iron accumulation in endometriosis. Hum Reprod. 2021 Mar 18;36(4):951-964.

[2]. Erastin Disrupts Mitochondrial Permeability Transition Pore (mPTP) and Induces Apoptotic Death of Colorectal Cancer Cells. PLoS One. 2016 May 12;11(5):e0154605.

[3]. Ferroptosis: process and function. Cell Death Differ. 2016 Mar;23(3):369-79.

[4]. MT1DP loaded by folate-modified liposomes sensitizes erastin-induced ferroptosis via regulating miR-365a-3p/NRF2 axis in non-small cell lung cancer cells. Cell Death Dis. 2020 Sep 14;11(9):751.

[5]. Piperlongumine Inhibits Thioredoxin Reductase 1 by Targeting Selenocysteine Residues and Sensitizes Cancer Cells to Erastin. Antioxidants (Basel). 2022 Apr 4;11(4):710.

[6]. Activation of the reverse transsulfuration pathway through NRF2/CBS confers erastin-induced ferroptosis resistance. Br J Cancer. 2020 Jan;122(2):279-292.

[7]. Mitochondrial fission links ECM mechanotransduction to metabolic redox homeostasis and metastatic chemotherapy resistance. Nat Cell Biol. 2022 Feb;24(2):168-180.

[8]. Mitochondrial transplantation rescues neuronal cells from ferroptosis. Free Radic Biol Med. 2023 Nov 1;208:62-72.

[9]. Ibrutinib facilitates the sensitivity of colorectal cancer cells to ferroptosis through BTK/NRF2 pathway. Cell Death Dis. 2023 Feb 23;14(2):151.

[10]. The Tumor Suppressor p53 Limits Ferroptosis by Blocking DPP4 Activity. Cell Rep. 2017 Aug 15;20(7):1692-1704.

Erastin is a member of the class of quinazolines that is quinazolin-4(3H)-one in which the hydrogens at positions 2 and 3 are replaced by 1-{4-[(4-chlorophenoxy)acetyl]piperazin-1-yl}ethyl and 2-ethoxyphenyl groups, respectively. It is an inhibitor of voltage-dependent anion-selective channels (VDAC2 and VDAC3) and a potent ferroptosis inducer. It has a role as a ferroptosis inducer, an antineoplastic agent and a voltage-dependent anion channel inhibitor. It is a member of quinazolines, a member of monochlorobenzenes, an aromatic ether, a N-acylpiperazine, a N-alkylpiperazine, a diether and a tertiary carboxamide.

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Study question: Could erastin activate ferroptosis to regress endometriotic lesions?
Summary answer: Erastin could induce ferroptosis to regress endometriotic lesions in endometriosis.
What is known already: Ectopic endometrial stromal cells (EESCs) are in an iron overloading microenvironment and tend to be more sensitive to oxidative damage. The feature of erastin-induced ferroptosis is iron-dependent accumulation of lethal lipid reactive oxygen species (ROS).
Study design, size, duration: Eleven patients without endometriosis and 21 patients with endometriosis were recruited in this study. Primary normal and ectopic endometrial stromal cells were isolated, cultured and subjected to various treatments. The in vivo study involved 10 C57BL/6 female mice to establish the model of endometriosis.
Participants/materials, setting, methods: The markers of ferroptosis were assessed by cell viability, lipid peroxidation level and morphological changes. The cell viability was measured by colorimetric method, lipid peroxidation levels were measured by flow cytometry, and morphological changes were observed by transmission electron microscopy. Immunohistochemistry and western blot were used to detect ferroportin (FPN) expression. Prussian blue staining and immunofluorescent microscopy of catalytic ferrous iron were semi-quantified the levels of iron. Adenovirus-mediated overexpression and siRNA-mediated knockdown were used to investigate the role of FPN on erastin-induced ferroptosis in EESCs.
Main results and the role of chance: EESCs were more susceptible to erastin treatment, compared to normal endometrial stromal cells (NESCs) (P<0.05). Treatment of cultured EESCs with erastin dramatically increased the total ROS level (P<0.05, versus control), lipid ROS level (P<0.05, versus NESCs) and intracellular iron level (P<0.05, versus NESCs). The cytotoxicity of erastin could be attenuated by iron chelator, deferoxamine (DFO), and ferroptosis inhibitors, ferrostatin-1 and liproxstatin-1, (P<0.05, versus erastin) in EESCs. In EESCs with erastin treatment, shorter and condensed mitochondria were observed by electron microscopy. These findings together suggest that erastin is capable to induce EESC death by ferroptosis. However, the influence of erastin on NESCs was slight. The process of erastin-induced ferroptosis in EESCs accompanied iron accumulation and decreased FPN expression. The overexpression of FPN ablated erastin-induced ferroptosis in EESCs. In addition, knockdown of FPN accelerated erastin-induced ferroptosis in EESCs. In a mouse model of endometriosis, we found ectopic lesions were regressed after erastin administration.
Large scale data: N/A.
Limitations, reasons for caution: This study was mainly conducted in primary human endometrial stromal cells. Therefore, the function of FPN in vivo need to be further investigated.
Wider implications of the findings: Our findings reveal that erastin may serve as a potential therapeutic treatment for endometriosis.
Study funding/competing interest(s): This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors. The authors declare no conflict of interest.[1]

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We here evaluated the potential anti-colorectal cancer activity by erastin, a voltage-dependent anion channel (VDAC)-binding compound. Our in vitro studies showed that erastin exerted potent cytotoxic effects against multiple human colorectal cancer cell lines, possibly via inducing oxidative stress and caspase-9 dependent cell apoptosis. Further, mitochondrial permeability transition pore (mPTP) opening was observed in erastin-treated cancer cells, which was evidenced by VDAC-1 and cyclophilin-D (Cyp-D) association, mitochondrial depolarization, and cytochrome C release. Caspase inhibitors, the ROS scavenger MnTBAP, and mPTP blockers (sanglifehrin A, cyclosporin A and bongkrekic acid), as well as shRNA-mediated knockdown of VDAC-1, all significantly attenuated erastin-induced cytotoxicity and apoptosis in colorectal cancer cells. On the other hand, over-expression of VDAC-1 augmented erastin-induced ROS production, mPTP opening, and colorectal cancer cell apoptosis. In vivo studies showed that intraperitoneal injection of erastin at well-tolerated doses dramatically inhibited HT-29 xenograft growth in severe combined immunodeficient (SCID) mice. Together, these results demonstrate that erastin is cytotoxic and pro-apoptotic to colorectal cancer cells. Erastin may be further investigated as a novel anti-colorectal cancer agent. [2]

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Piperlongumine, a natural alkaloid substance extracted from the fruit of the long pepper (Piper longum Linn.), is known to inhibit the cytosolic thioredoxin reductase (TXNRD1 or TrxR1) and selectively kill cancer cells. However, the details and mechanism of the inhibition by piperlongumine against TXNRD1 remain unclear. In this study, based on the classical DTNB reducing assay, irreversible inhibition of recombinant TXNRD1 by piperlongumine was found and showed an apparent kinact value of 0.206 × 10-3 µM-1 min-1. Meanwhile, compared with the wild-type TXNRD1 (-GCUG), the UGA-truncated form (-GC) of TXNRD1 was resistant to piperlongumine, suggesting the preferential target of piperlongumine is the selenol (-SeH) at the C-terminal redox motif of the enzyme. Interestingly, the high concentration of piperlongumine-inhibited TXNRD1 showed that its Sec-dependent activity is decayed but its intrinsic NADPH oxidase activity is retained. Furthermore, piperlongumine did not induce ferroptosis in HCT116 cells at 10 µM, whereas significantly promoted erastin-induced lipid oxidation, which could be alleviated by supplying glutathione (GSH) or N-acetyl L-cysteine (NAC). However, restricting GSH synthesis by inhibiting glutaminase (GLS) using the small molecule inhibitor CB-839 only slightly enhanced erastin-induced cell death. Taken together, this study elucidates the molecular mechanism of the antitumor capacity of piperlongumine by targeting TXNRD1 and reveals the potential possibility of inhibiting TXNRD1 to strengthen cancer cells' ferroptosis. [5]

C30H31CLN4O4

547.04

546.203

C, 65.87; H, 5.71; Cl, 6.48; N, 10.24; O, 11.70

571203-78-6

11214940

White to off-white solid

1.3±0.1 g/cm3

721.9±70.0 °C at 760 mmHg

390.4±35.7 °C

0.0±2.3 mmHg at 25°C

1.634

4.75

76.9

0

6

8

39

871

0

ClC1C([H])=C([H])C(=C([H])C=1[H])OC([H])([H])C(N1C([H])([H])C([H])([H])N(C([H])([H])C1([H])[H])C([H])(C([H])([H])[H])C1=NC2=C([H])C([H])=C([H])C([H])=C2C(N1C1=C([H])C([H])=C([H])C([H])=C1OC([H])([H])C([H])([H])[H])=O)=O

BKQFRNYHFIQEKN-UHFFFAOYSA-N

InChI=1S/C30H31ClN4O4/c1-3-38-27-11-7-6-10-26(27)35-29(32-25-9-5-4-8-24(25)30(35)37)21(2)33-16-18-34(19-17-33)28(36)20-39-23-14-12-22(31)13-15-23/h4-15,21H,3,16-20H2,1-2H3

2-(1-(4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)ethyl)-3-(2-ethoxyphenyl)quinazolin-4(3H)-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 该产品在溶液状态不稳定，请现配现用。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 1.25 mg/mL (2.29 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 12.5 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 2 中的溶解度: ≥ 1 mg/mL (1.83 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 10.0 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 3 中的溶解度: 5% DMSO+corn oil: 2.5mg/mL

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配方 4 中的溶解度: 5 mg/mL (9.14 mM) in 50% PEG300 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
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10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
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