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Purity: >98.00%

实验参考方法

Cell experiment [1]:
Cell lines	HEK293 cells
Preparation Method	No-tagged MS4A12 and GCaMP-MS4A12 plasmids were mixed at a 1:1 ratio and then transfected into HEK293 cells grown on coverslips.Thapsigargin of 5 μM was used for stimulation and store depletion, after which 5 mM Ca2⁺ regular solution was perfused to visualize Ca2⁺ influx-induced fluorescence of GCaMP-MS4A12.
Reaction Conditions	5 μM; 37°C
Applications	When cells were stimulated with a store-depleting condition (5 μM thapsigargin for 5 min in the 5 mM Ca2⁺ extracellular solution), the fluorescence of GCaMP-M12 very rapidly and greatly increased.
Animal experiment [2]:
Animal models	6- to 8-week-old BALB/c mice (female)
Preparation Method	The mice were intranasally infected with 3 MLD50 of PR8/H1N1 virus. Twelve hours post-infection, Thapsigargin (1.5 μg/kg/day), oseltamivir (45 mg/kg/day) or PBS+DMSO (percentage DMSO in PBS-DMSO control equal to other treatments) was orally administered by gavage daily for 5 days. At 3 and 5 days post-infection, the lungs of four mice from each group were collected for viral titration.
Dosage form	1.5 μg/kg/day; p.o.
Applications	The Thapsigargin-treated group showed significantly improved survival, reduced virus shedding at 3 and 5 days post-infection and less severe weight loss.
References: [1]. Han JW, et al. Plasma Membrane Localized GCaMP-MS4A12 by Orai1 Co-Expression Shows Thapsigargin- and Ca2⁺-Dependent Fluorescence Increases. Mol Cells. 2021 Apr 30;44(4):223-232. [2]. Al-Beltagi S, et al. Thapsigargin Is a Broad-Spectrum Inhibitor of Major Human Respiratory Viruses: Coronavirus, Respiratory Syncytial Virus and Influenza A Virus. Viruses. 2021 Feb 3;13(2):234.

产品描述

Thapsigargin is an inhibitor of the sarcoplasmic/endoplasmic reticulum Ca2⁺ ATPase pump.^[1] Thapsigargin can induce a potent host antiviral response that blocks influenza A virus replication at nano-molar non-cytotoxic levels.^[2]

In vitro efficacy test it shown that HEp2 cells primed with 0.5 µM Thapsigargin (as a non-cytotoxic inhibitor) before infection reduced progeny virus production by almost 10,000-fold. A549 cells primed with 0.3 µM Thapsigargin, in turn, was more inhibitory than the RDV-treated cells by 450-fold.^[3] In vitro, treatment with 10 μM of Thapsigargin (a specific blocker SERCAs), can null EMF response by impairing the replenishment of the ER.^[4] In vitro test it indicated that treatment with 0.001 μM - 1 μM of thapsigargin may arrest cell proliferations in MH7A human rheumatoid arthritis synovial cells in a time- and dose-dependent manner.^[5] Treatment with 1 μM of thapsigargin mediated sensitization to TRAIL in ESCC cell lines through the activation of AMPK from the aspects of protein function and existence.^[7]

In vivo efficacy study it demonstrated that treatment with 0.5ug/g/body weight of Thapsigargin was safe and did not elicit any adverse effects on survival in mice.^[6] In vivo, mice were treated with 1 mg/kg thapsigargin and 60 mg/kg hrTRAIL intraperitoneally for five times per week, improved the expression of CHOP, increased AMPK phosphorylation, increased the expression of DR5, increased the expression of Bax, and activated Caspase 9, while suppressing Bcl2 expression.^[7]

他普西加林是一种肌浆网/内质网Ca2+ ATP酶泵的抑制剂。他普西加林可以在纳摩尔非细胞毒性水平下诱导强效宿主抗病毒反应，阻止流感A病毒的复制。

实验室内的有效性测试表明，在感染前使用0.5微米的硫酸沙脂（作为非细胞毒性抑制剂）预处理HEp2细胞可以将后代病毒产量减少近10,000倍。相反，使用0.3微米硫酸沙脂预处理A549细胞比RDV治疗的细胞更具抑制作用，可使后代病毒产量降低450倍。在体外实验中，使用10微米的硫酸沙脂（一种特定的SERCAs阻断剂），可以通过影响内质网再生来消除EMF响应。体外测试表明，以时间和剂量依赖方式处理MH7A人类类风湿关节炎滑液细胞时，使用0.001-1微米硫酸沙脂可能会阻止其增殖。在ESCc细胞系中，以1微米硫酸沙脂处理介导了对TRAIL敏化作用，并通过AMPK激活从蛋白功能和存在方面进行了验证。

在体内疗效研究中，使用0.5ug/g/体重的Thapsigargin治疗是安全的，并且不会对小鼠的生存产生任何不良影响。在活体实验中，小鼠每周接受1mg/kg Thapsigargin和60mg/kg hrTRAIL腹腔注射五次，在表达CHOP、增加AMPK磷酸化、增加DR5表达、增加Bax表达和激活Caspase 9的同时，抑制了Bcl2的表达。

References:
[1].Lytton J., Westlin M., Hanley M.R. Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. J. Biol. Chem. 1991;266:17067–17071.
[2].Goulding L.V., Yang J., Jiang Z., Zhang H., Lea D., Emes R.D., Dottorini T., Pu J., Liu J., Chang K.C. Thapsigargin at non-cytotoxic levels induces a potent host antiviral response that blocks influenza A virus replication. Viruses. 2020;12:1093.
[3].Al-Beltagi S, et al. Thapsigargin Is a Broad-Spectrum Inhibitor of Major Human Respiratory Viruses: Coronavirus, Respiratory Syncytial Virus and Influenza A Virus. Viruses. 2021 Feb 3;13(2):234.
[4].Bertagna F, et al. Thapsigargin blocks electromagnetic field-elicited intracellular Ca2⁺ increase in HEK 293 cells. Physiol Rep. 2022 May;10(9):e15189.
[5].Wang H, et al. Effects of thapsigargin on the proliferation and survival of human rheumatoid arthritis synovial cells. ScientificWorldJournal. 2014 Feb 9;2014:605416.
[6].Abdullahi A, et al. Modeling Acute ER Stress in Vivo and in Vitro. Shock. 2017 Apr;47(4):506-513.
[7].Ma Z, et al. Thapsigargin sensitizes human esophageal cancer to TRAIL-induced apoptosis via AMPK activation. Sci Rep. 2016 Oct 12;6:35196.

Chemical Properties

Cas No.	67526-95-8	SDF
别名	毒胡萝卜素
化学名	(3S,3aR,4S,6S,6aR,7S,8S,9bS)-6-acetoxy-4-(butyryloxy)-3,3a-dihydroxy-3,6,9-trimethyl-8-(((Z)-2-methylbut-2-enoyl)oxy)-2-oxo-2,3,3a,4,5,6,6a,7,8,9b-decahydroazuleno[4,5-b]furan-7-yl octanoate
Canonical SMILES	O[C@@]([C@H]1OC(CCC)=O)([C@]2(C)O)[C@H](C([C@H]([C@@H]3OC(CCCCCCC)=O)[C@@](C1)(C)OC(C)=O)=C(C)[C@@H]3OC(/C(C)=C\C)=O)OC2=O
分子式	C₃₄H₅₀O₁₂	分子量	650.76
溶解度	30mg/ml in ethanol, DMSO, DMF	储存条件	Store at -20°C, sealed storage, away from moisture and light
General tips	请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。
Shipping Condition	评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。

溶解性数据

制备储备液
		1 mg	5 mg	10 mg
	1 mM	1.5367 mL	7.6833 mL	15.3666 mL
	5 mM	0.3073 mL	1.5367 mL	3.0733 mL
	10 mM	0.1537 mL	0.7683 mL	1.5367 mL

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动物体内配方计算器 (澄清溶液)

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每只动物给药体积

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工作液浓度： mg/ml；

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体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。

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Research Update

Thapsigargin-From Traditional Medicine to Anticancer Drug

A sesquiterpene lactone, thapsigargin, is a phytochemical found in the roots and fruits of Mediterranean plants from Thapsia L. species that have been used for centuries in folk medicine to treat rheumatic pain, lung diseases, and female infertility. More recently thapsigargin was found to be a potent cytotoxin that induces apoptosis by inhibiting the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA) pump, which is necessary for cellular viability. This biological activity encouraged studies on the use of thapsigargin as a novel antineoplastic agent, which were, however, hampered due to high toxicity of this compound to normal cells. In this review, we summarized the recent knowledge on the biological activity and molecular mechanisms of thapsigargin action and advances in the synthesis of less-toxic thapsigargin derivatives that are being developed as novel anticancer drugs.

Thapsigargin: key to new host-directed coronavirus antivirals?

Despite the great success of vaccines that protect against RNA virus infections, and the development and clinical use of a limited number of RNA virus-specific drugs, there is still an urgent need for new classes of antiviral drugs against circulating or emerging RNA viruses. To date, it has proved difficult to efficiently suppress RNA virus replication by targeting host cell functions, and there are no approved drugs of this type. This opinion article discusses the recent discovery of a pronounced and sustained antiviral activity of the plant-derived natural compound thapsigargin against enveloped RNA viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Middle East respiratory syndrome coronavirus (MERS-CoV), and influenza A virus. Based on its mechanisms of action, thapsigargin represents a new prototype of compounds with multimodal host-directed antiviral activity.

Targeting thapsigargin towards tumors

The skin irritating principle from Thapsia garganica was isolated, named thapsigargin and the structure elucidated. By inhibiting the sarco/endoplasmic reticulum Ca(2+) ATPase (SERCA) thapsigargin provokes apoptosis in almost all cells. By conjugating thapsigargin to peptides, which are only substrates for either prostate specific antigen (PSA) or prostate specific membrane antigen (PSMA) prodrugs were created, which selectively affect prostate cancer cells or neovascular tissue in tumors. One of the prodrug is currently tested in clinical phase II. The prodrug under clinical trial has been named mipsagargin.

Endoplasmic reticulum stress induced by tunicamycin and thapsigargin protects against transient ischemic brain injury: Involvement of PARK2-dependent mitophagy

Transient cerebral ischemia leads to endoplasmic reticulum (ER) stress. However, the contributions of ER stress to cerebral ischemia are not clear. To address this issue, the ER stress activators tunicamycin (TM) and thapsigargin (TG) were administered to transient middle cerebral artery occluded (tMCAO) mice and oxygen-glucose deprivation-reperfusion (OGD-Rep.)-treated neurons. Both TM and TG showed significant protection against ischemia-induced brain injury, as revealed by reduced brain infarct volume and increased glucose uptake rate in ischemic tissue. In OGD-Rep.-treated neurons, 4-PBA, the ER stress releasing mechanism, counteracted the neuronal protection of TM and TG, which also supports a protective role of ER stress in transient brain ischemia. Knocking down the ER stress sensor Eif2s1, which is further activated by TM and TG, reduced the OGD-Rep.-induced neuronal cell death. In addition, both TM and TG prevented PARK2 loss, promoted its recruitment to mitochondria, and activated mitophagy during reperfusion after ischemia. The neuroprotection of TM and TG was reversed by autophagy inhibition (3-methyladenine and Atg7 knockdown) as well as Park2 silencing. The neuroprotection was also diminished in Park2(+/-) mice. Moreover, Eif2s1 and downstream Atf4 silencing reduced PARK2 expression, impaired mitophagy induction, and counteracted the neuroprotection. Taken together, the present investigation demonstrates that the ER stress induced by TM and TG protects against the transient ischemic brain injury. The PARK2-mediated mitophagy may be underlying the protection of ER stress. These findings may provide a new strategy to rescue ischemic brains by inducing mitophagy through ER stress activation.

Thapsigargin--from Thapsia L. to mipsagargin

The sesquiterpene lactone thapsigargin is found in the plant Thapsia garganica L., and is one of the major constituents of the roots and fruits of this Mediterranean species. In 1978, the first pharmacological effects of thapsigargin were established and the full structure was elucidated in 1985. Shortly after, the overall mechanism of the Sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) inhibition that leads to apoptosis was discovered. Thapsigargin has a potent antagonistic effect on the SERCA and is widely used to study Ca2+-signaling. The effect on SERCA has also been utilized in the treatment of solid tumors. A prodrug has been designed to target the blood vessels of cancer cells; the death of these blood vessels then leads to tumor necrosis. The first clinical trials of this drug were initiated in 2008, and the potent drug is expected to enter the market in the near future under the generic name Mipsagargin (G-202). This review will describe the discovery of the new drug, the on-going elucidation of the biosynthesis of thapsigargin in the plant and attempts to supply the global market with a novel potent anti-cancer drug.

Thapsigargin

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