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Purity: >99.50%

实验参考方法

Kinase experiment [1]:
Preparation Method	For studies of the kinetics of inhibition of PtdIns-3-kinase by wortmannin, bovine brain enzyme was incubated with 0 to 14 nM wortmannin with varying ATP concentrations from 2.5 to 20 μM. For studies of the time course of PtdIns-3-kinase inhibition, wortmannin 0 to 14 nM, enzyme, and PtdIns were incubated for various times at room temperature before adding [γ-32P]ATP to start the reaction.
Reaction Conditions	0-14 nM wortmannin
Applications	Kinetic analysis demonstrates that wortmannin is a noncompetitive, irreversible inhibitor of phosphatidylinositol-3-kinase, with inactivation being both time- and concentration-dependent.
Cell experiment [2]:
Cell lines	Jurkat cells
Preparation Method	Jurkat-soluble fractions were treated with AX7503 (50 nM) for 60 min with or without preincubation with various concentrations of wortmannin (0.25‿0 nM) or LY294002 (10‿0,000 nM) for 10 min.
Reaction Conditions	Wortmannin (0.25‿0 nM) for 10 min
Applications	PLK1 activity was inhibited by wortmannin with an IC 50 value of 24 nM.
Animal experiment [3]:
Animal models	Scid mice
Preparation Method	Oral gavage; daily; in Scid mice; one group of eight mice is dosed with Wortmannin 1 mg/kg for all 14 days. The second group of eight mice is dosed with Wortmannin 1.5 mg/kg for the first 5 days and the dose is decreased to 1 mg/kg for the remaining treatment period.
Dosage form	1 mg/kg for 14 days; 1.5 mg/kg for 5 days + 1.0 mg/kg for 9 days.
Applications	Wortmannin treatment significantly slower the growth rate of murine C3H mammary tumor and human MCF-7 breast cancer xenograft. A dose of 1 mg/kg Wortmannin for 7 days decrease the tumor burdens in mice with established murine C3H mammary tumors by 54% relative to controls. Human MCF-7 breast cancer xenograft burdens are decreased by 97% relative to controls after 14 days of 1 mg/kg Wortmannin beginning 1 day after tumor implantation.
References: [1]. Powis G, Bonjouklian R, et al. Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3-kinase. Cancer Res. 1994 May 1;54(9):2419-23. PMID: 8162590. [2]. Liu Y, Shreder KR, et al. Wortmannin, a widely used phosphoinositide 3-kinase inhibitor, also potently inhibits mammalian polo-like kinase. Chem Biol. 2005 Jan;12(1):99-107. doi: 10.1016/j.chembiol.2004.11.009. PMID: 15664519. [3]. Lemke LE, Paine-Murrieta GD, et al. Wortmannin inhibits the growth of mammary tumors despite the existence of a novel wortmannin-insensitive phosphatidylinositol-3-kinase. Cancer Chemother Pharmacol. 1999;44(6):491-7. doi: 10.1007/s002800051123. PMID: 10550570.

产品描述

Wortmannin is a highly potent direct inhibitor of PI3-kinase specificity originally derived from fungi (1,2). This inhibition is irreversible and non-competitive with an IC50 of 3 nM^[2]^[3]. Wortmannin does not inhibit PI4 kinase, protein kinase C, or protein tyrosine kinases^[4].

In Jurkat cell, at the concentrations of wortmannin commonly used to inhibit PI 3-kinases, PLK1 is also significantly inhibited^[5]. In Swiss 3T3 cells, Wortmannin is a selective and reversible phosphatidylinositol 3-kinase inhibitor with an IC50 value of 1.9 nM^[4].PI3-kinase is involved in the signal transduction pathway responsible for histamine secretion following stimulation of Fc epsilon RI and that wortmannin blocks these responses through direct interaction with the catalytic subunit of this enzyme^[1].

Wortmannin treatment significantly slower the growth rate of murine C3H mammary tumor and human MCF-7 breast cancer xenograft. A dose of 1 mg/kg Wortmannin for 7 days decrease the tumor burdens in mice with established murine C3H mammary tumors by 54% relative to controls. Human MCF-7 breast cancer xenograft burdens are decreased by 97% relative to controls after 14 days of 1 mg/kg Wortmannin beginning 1 day after tumor implantation^[6]. Wortmannin inhibits myosin light chain phosphorylation and contraction in rat aorta. As an inhibitor of MLCK, wortmannin can be used as a vasodilator and anti-inflammatory agent^[7].

References:
[1]: Yano H, Nakanishi S, et,al. Inhibition of histamine secretion by wortmannin through the blockade of phosphatidylinositol 3-kinase in RBL-2H3 cells. J Biol Chem. 1993 Dec 5;268(34):25846-56. PMID: 7503989.
[2]: Moon EK, Kim SH, et,al. Autophagy inhibitors as a potential antiamoebic treatment for Acanthamoeba keratitis. Antimicrob Agents Chemother. 2015 Jul;59(7):4020-5. doi: 10.1128/AAC.05165-14. Epub 2015 Apr 20. PMID: 25896709; PMCID: PMC4468686.
[3]: Liu Y, Jiang N, et,al. Polo-like kinases inhibited by wortmannin. Labeling site and downstream effects. J Biol Chem. 2007 Jan 26;282(4):2505-11. doi: 10.1074/jbc.M609603200. Epub 2006 Nov 29. PMID: 17135248.
[4]: Powis G, Bonjouklian R, et,al. Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3-kinase. Cancer Res. 1994 May 1;54(9):2419-23. PMID: 8162590.
[5]: Liu Y, Shreder KR, et,al. Wortmannin, a widely used phosphoinositide 3-kinase inhibitor, also potently inhibits mammalian polo-like kinase. Chem Biol. 2005 Jan;12(1):99-107. doi: 10.1016/j.chembiol.2004.11.009. PMID: 15664519.
[6]: Lemke LE, Paine-Murrieta GD, et,al. Wortmannin inhibits the growth of mammary tumors despite the existence of a novel wortmannin-insensitive phosphatidylinositol-3-kinase. Cancer Chemother Pharmacol. 1999;44(6):491-7. doi: 10.1007/s002800051123. PMID: 10550570.
[7]: Nakanishi S, Kakita S, et,al.Wortmannin, a microbial product inhibitor of myosin light chain kinase. J Biol Chem. 1992 Feb 5;267(4):2157-63. PMID: 1733924.

渥曼青霉素是一种高效的 PI3 激酶特异性直接抑制剂，最初源自真菌 (1,2)。这种抑制是不可逆的和非竞争性的，IC50 为 3 nM^[2]^[3]。渥曼青霉素不抑制 PI4 激酶、蛋白激酶 C 或蛋白酪氨酸激酶^[4]。

在 Jurkat 细胞中，在通常用于抑制 PI 3-激酶的渥曼青霉素浓度下, PLK1 也被显着抑制^[5]。在 Swiss 3T3 细胞中，Wortmannin 是一种选择性和可逆的磷脂酰肌醇 3-激酶抑制剂，IC50 值为 1.9 nM^[4]。PI3-激酶参与刺激后组胺分泌的信号转导通路Fc epsilon RI 和渥曼青霉素通过与该酶的催化亚基直接相互作用来阻断这些反应^[1]。

渥曼青霉素治疗显着减缓了小鼠 C3H 乳腺肿瘤的生长速度和人类 MCF-7 乳腺癌异种移植物。相对于对照组，以 1 mg/kg 的剂量服用 Wortmannin 7 天可使患有小鼠 C3H 乳腺肿瘤的小鼠的肿瘤负荷降低 54%。在肿瘤植入后 1 天开始使用 1 mg/kg 渥曼青霉素 14 天后，人类 MCF-7 乳腺癌异种移植物负荷相对于对照组降低了 97%^[6]。 Wortmannin 抑制大鼠主动脉肌球蛋白轻链磷酸化和收缩。作为MLCK抑制剂，渥曼青霉素可作为血管扩张剂和抗炎剂使用^[7]。

Chemical Properties

Cas No.	19545-26-7	SDF
别名	渥曼青霉素; SL-2052; KY-12420
化学名	(1S,6bR,9aS,11R,11bR)-1-(methoxymethyl)-9a,11b-dimethyl-3,6,9-trioxo-3,6,6b,7,8,9,9a,10,11,11b-decahydro-1H-furo[4,3,2-de]indeno[4,5-h]isochromen-11-yl acetate
Canonical SMILES	O=C1[C@](C([H])([H])[C@@]2([H])OC(C([H])([H])[H])=O)(C([H])([H])[H])[C@](C([H])([H])C1([H])[H])([H])C(C3=O)=C2[C@]4(C([H])([H])[H])C5=C3OC([H])=C5C(O[C@]4([H])C([H])([H])OC([H])([H])[H])=O
分子式	C₂₃H₂₄O₈	分子量	428.43
溶解度	≥ 21.42mg/mL in DMSO	储存条件	Store at -20°C
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	2.3341 mL	11.6705 mL	23.341 mL
	5 mM	0.4668 mL	2.3341 mL	4.6682 mL
	10 mM	0.2334 mL	1.1671 mL	2.3341 mL

摩尔浓度计算器
稀释计算器
分子量计算器

计算

动物体内配方计算器 (澄清溶液)

第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量）

给药剂量

mg/kg

动物平均体重

g

每只动物给药体积

ul

动物数量

只

第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方）

% DMSO+ % + % Tween 80+ % saline

计算重置

计算结果:

工作液浓度： mg/ml；

DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，注：如该浓度超过该批次药物DMSO溶解度，请先联系GLPBIO）；

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。

注意：1. 首先保证母液是澄清的；
2. 一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。

Research Update

Wortmannin, a specific inhibitor of phosphatidylinositol-3-kinase, induces accumulation of DNA double-strand breaks

Wortmannin, a fungal metabolite, is a specific inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, which includes double-stranded DNA dependent protein kinase (DNA-PK) and ataxia telangiectasia mutated kinase (ATM). We investigated the effects of wortmannin on DNA damage in DNA-PK-deficient cells obtained from severe combined immunodeficient mice (SCID cells). Survival of wortmannin-treated cells decreased in a concentration-dependent manner. After treatment with 50 μM wortmannin, survival decreased to 60% of that of untreated cells. We observed that treatment with 20 and 50 μM wortmannin induced DNA damage equivalent to that by 0.37 and 0.69 Gy, respectively, of γ-ray radiation. The accumulation of DNA double-strand breaks (DSBs) in wortmannin-treated SCID cells was assessed using pulsed-field gel electrophoresis. The maximal accumulation was observed 4 h after treatment. Moreover, the presence of DSBs was confirmed by the ability of nuclear extracts from γ-ray-irradiated SCID cells to produce in vitro phosphorylation of histone H2AX. These results suggest that wortmannin induces cellular toxicity by accumulation of spontaneous DSBs through inhibition of ATM.

Wortmannin as a unique probe for an intracellular signalling protein, phosphoinositide 3-kinase

Wortmannin is a fungal metabolite that so far has been shown to act as a selective inhibitor of phosphoinositide 3-kinase. It can therefore be used to investigate the convergence between two major cellular signalling systems: those involving G-protein-coupled receptors and those involving receptor tyrosine kinases. Importantly, wortmannin can enter intact cells, making whole-cell studies of the above signalling pathways possible.

Investigating the Inhibitory Effect of Wortmannin in the Hotspot Mutation at Codon 1047 of PIK3CA Kinase Domain: A Molecular Docking and Molecular Dynamics Approach

Oncogenic mutations in phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) are the most frequently reported in association with various forms of cancer. Several studies have reported the significance of hotspot mutations in a catalytic subunit of PIK3CA in association with breast cancer. Mutations are frequently observed in the highly conserved region of the kinase domain (797-1068 amino acids) of PIK3CA are activating or gain-of-function mutations. Mutation in codon 1047 occurs in the C-terminal region of the kinase domain with histidine (H) replaced by arginine (R), lysine (L), and tyrosine (Y). Pathogenicity and protein stability predictors PhD-SNP, Align GVGD, HANSA, iStable, and MUpro classified H1047R as highly deleterious when compared to H1047L and H1047Y. To explore the inhibitory activity of Wortmannin toward PIK3CA, the three-dimensional structure of the mutant protein was determined using homology modeling followed by molecular docking and molecular dynamics analysis. Docking studies were performed for the three mutants and native with Wortmannin to measure the differences in their binding pattern. Comparative docking study revealed that H1047R-Wortmannin complex has a higher number of hydrogen bonds as well as the best binding affinity next to the native protein. Furthermore, 100 ns molecular dynamics simulation was initiated with the docked complexes to understand the various changes induced by the mutation. Though Wortmannin was found to nullify the effect of H1047R over the protein, further studies are required for designing a better compound. As SNPs are major genetic variations observed in disease condition, personalized medicine would provide enhanced drug therapy.

Wortmannin targeting phosphatidylinositol 3-kinase suppresses angiogenic factors in shear-stressed endothelial cells

Modifications on shear stress-based mechanical forces are associated with pathophysiological susceptibility and their effect on endothelial cells (EC) needs to be better addressed looking for comprehending the cellular and molecular mechanisms. This prompted us to better evaluate the effects of shear stress in human primary venous EC obtained from the umbilical cord, using an in vitro model to mimic the laminar blood flow, reaching an intensity 1-4 Pa. First, our data shows there is a significant up-expression of phosphatidylinositol 3-kinase (PI3K) in shear-stressed cells culminating downstream with an up-phosphorylation of AKT and up-expression of MAPK-ERK, concomitant to a dynamic cytoskeleton rearrangement upon integrin subunits (α4 and ? 3) requirements. Importantly, the results show there is significant involvement of nitric oxide synthase (eNOS), nNOS, and vascular endothelial growth factors receptor 2 (VEGFR2) in shear-stressed EC, while cell cycle-related events seem to being changed. Additionally, although diminution of 5-hydroxymethylcytosine in shear-stressed EC, suggesting a global repression of genes transcription, the promoters of PI3K and eNOS genes were significantly hydroxymethylated corroborating with their respective transcriptional profiles. Finally, to better address, the pivotal role of PI3K in shear-stressed EC we have revisited these biological issues by wortmannin targeting PI3K signaling and the data shows a dependency of PI3K signaling in controlling the expression of VGFR1, VGFR2, VEGF, and eNOS, once these genes were significantly suppressed in the presence of the inhibitor, as well as transcripts from Ki67 and CDK2 genes. Finally, our data still shows a coupling between PI3K and the epigenetic landscape of shear-stressed cells, once wortmannin promotes a significant suppression of ten-11 translocation 1 (TET1), TET2, and TET3 genes, evidencing that PI3K signaling is a necessary upstream pathway to modulate TET-related genes. In this study we determined the major mechanotransduction pathway by which blood flow driven shear stress activates PI3K which plays a pivotal role on guaranteeing endothelial cell phenotype and vascular homeostasis, opening novel perspectives to understand the molecular basis of pathophysiological disorders related with the vascular system.

Wortmannin and Wortmannine Analogues from an Undescribed Niesslia sp

In the course of our studies of coprophilous fungi as sources of antifungal agents, a strain of an undescribed species in the genus Niesslia (TTI-0426) was isolated from horse dung collected in Texas. An extract from fermentation cultures of this strain afforded two new antifungal wortmannin derivatives, wortmannins C and D (1 and 2), as well as four additional new related compounds, wortmannines B1-B4 (3-6), containing an unusual ring system. The structures of these metabolites were established mainly by analysis of HRESIMS and 2D NMR data. Relative configurations were assigned using NOESY data, and the structure assignments were supported by NMR comparison with similar compounds. Wortmannins C and D showed activity against Cryptococcus neoformans and Candida albicans in disk assays, but low MIC potency observed for 1 was suggested to be due in part to efflux processes on the basis of assay results for a Schizosaccharomyces pombe efflux mutant in comparison to wild-type.

Wortmannin

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