Pironetin
(Synonyms: NK 10958, NL 9C, PA 48153c, (–)-Pironetin) 目录号 : GC38456
An inhibitor of microtubule assembly
Cas No.:151519-02-7
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Pironetin is a bacterial metabolite originally isolated from Streptomyces that has diverse biological activities, including anti-proliferative, immunosuppressive, and plant growth regulatory properties.1,2,3,4 It binds to tubulin with a Kd value of 0.33 μM and increases the critical concentration (CrC) for tubulin assembly in glycerol-assembling buffer (GAB) at a concentration of 25 μM.2,3 It also induces G2/M phase cell cycle arrest in 3Y1 rat fibroblasts and apoptosis in HL-60 human leukemia cells when used at concentrations of 50 ng/ml and 33 nM, respectively.2,5 It inhibits the growth of HT-29 human colorectal and MCF-7 human breast cancer cells (IC50s = 6.4 and 6 nM, respectively) but also of non-cancerous human HEK293 cells (IC50 = 17 nM). It also inhibits the growth of A2780 human ovarian carcinoma cells, as well as of the drug-resistant, P-glycoprotein-expressing A2780AD subline (IC50s = 8 and 25 nM, respectively). Pironetin (5 mg/kg) decreases the generation of cytotoxic T lymphocytes in mice in response to immunization by EL4 allogeneic mouse T lymphocytes.4 It also inhibits rice plant growth by 23% when applied nine days prior to heading.1
1.Kobayashi, S., Tsuchiya, K., Harada, T., et al.Pironetin, a novel plant growth regulator produced by Streptomyces sp. NK10958. I. Taxonomy, production, isolation and preliminary characterizationJ. Antibiot. (Tokyo)47(6)697-702(1994) 2.Kondoh, M., Usui, T., Nishikiori, T., et al.Apoptosis induction via microtubule disassembly by an antitumour compound, pironetinBiochem. J.340(Pt 2)411-416(1999) 3.Vilanova, C., Díaz-Oltra, S., Murga, J., et al.Design and synthesis of pironetin analogue/colchicine hybrids and study of their cytotoxic activity and mechanisms of interaction with tubulinJ. Med. Chem.57(24)10391-10403(2014) 4.Yasui, K., Tamura, Y., Nakatani, T., et al.Chemical modification of PA-48153C, a novel immunosuppressant isolated from Streptomyces prunicolor PA-48153J. Antibiot. (Tokyo)49(2)173-180(1996) 5.Kondoh, M., Usui, T., Kobayashi, S., et al.Cell cycle arrest and antitumor activity of pironetin and its derivativesCancer Lett.126(1)29-32(1998)
| Cas No. | 151519-02-7 | SDF | |
| 别名 | NK 10958, NL 9C, PA 48153c, (–)-Pironetin | ||
| Canonical SMILES | O=C1C=C[C@H]([C@H](O1)C[C@H]([C@@H]([C@@H]([C@H](C/C=C/C)C)OC)C)O)CC | ||
| 分子式 | C19H32O4 | 分子量 | 324.45 |
| 溶解度 | Dichloromethane: soluble,DMSO: soluble,Ethanol: soluble | 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。 |
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| 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。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.0821 mL | 15.4107 mL | 30.8214 mL |
| 5 mM | 0.6164 mL | 3.0821 mL | 6.1643 mL |
| 10 mM | 0.3082 mL | 1.5411 mL | 3.0821 mL |
| 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 |
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| % 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，
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。
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。
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Cytotoxicity of phenylpironetin analogs and the metabolic fate of Pironetin and phenylpironetin
Bioorg Chem 2022 Aug;125:105915.PMID:35660840DOI:10.1016/j.bioorg.2022.105915.
To improve Pironetin's metabolic stability we prepared four analogs by replacing its C12-14 segment with an aryl group. The antiproliferative activity of phenyl analog 4 was reduced two-fold and dihydroxy-4-fluorophenyl analog 5 was slightly more effective against OVCAR5 and A2780 ovarian cancer cell lines compared with the parent compound Pironetin (1). The activity of 4-fluorophenyl analog 6 was reduced 3-fold in both cell lines. The activity of 7-O-methyl analog 7 was reduced 36-fold in OVCAR5 cells and 47-fold and A2780 cells, compared with Pironetin. Phenylpironetin (4) was rapidly metabolized by mouse and human liver microsomes. We identified 17 human metabolites for phenyl analog 4 and 14 human metabolites for Pironetin. Metabolism occurred at the C12-13 moiety, the α,β-unsaturated lactone and the side chains of the molecules (C6-C11 segments). The significant extent of oxidative metabolism suggests that it may not be possible to attain a metabolically stable Pironetin analog by structural modifications of the parent compound.
Revisiting microtubule targeting agents: α-Tubulin and the Pironetin binding site as unexplored targets for cancer therapeutics
Bioorg Med Chem Lett 2019 Aug 1;29(15):1865-1873.PMID:31130264DOI:10.1016/j.bmcl.2019.05.042.
Molecules that bind to tubulin and disrupt tubulin dynamics are known as microtubule targeting agents. Treatment with a microtubule targeting agent leads to cell cycle arrest followed by apoptosis. Tubulin inhibitors have been highly effective in the clinical treatment of a variety of tumors and are being investigated for treatment of several other diseases. Currently, all FDA approved microtubule inhibitors bind to β-tubulin. Given the overall success of tubulin-binding agents in anticancer chemotherapy, α-tubulin is an attractive and unexplored target. Herein, we will discuss Pironetin, the only compound known to bind α-tubulin, with particular focus on the known biological properties, the total syntheses, exploration of its structure-activity relationship, and future directions.
Pironetin Binds Covalently to αCys316 and Perturbs a Major Loop and Helix of α-Tubulin to Inhibit Microtubule Formation
J Mol Biol 2016 Jul 31;428(15):2981-8.PMID:27395016DOI:10.1016/j.jmb.2016.06.023.
Microtubule-targeting agents are among the most powerful drugs used in chemotherapy to treat cancer patients. Pironetin is a natural product that displays promising anticancer properties by binding to and potently inhibiting tubulin assembly into microtubules; however, its molecular mechanism of action remained obscure. Here, we solved the crystal structure of the tubulin-pironetin complex and found that the compound covalently binds to Cys316 of α-tubulin. The structure further revealed that Pironetin perturbs the T7 loop and helix H8 of α-tubulin. Since both these elements are essential for establishing longitudinal tubulin contacts in microtubules, this result explains how Pironetin inhibits the formation of microtubules. Together, our data define the molecular details of the Pironetin binding site on α-tubulin and thus offer a promising basis for the rational design of Pironetin variants with improved activity profiles. They further extend our knowledge on strategies evolved by natural products to target and perturb the microtubule cytoskeleton.
Pironetin reacts covalently with cysteine-316 of α-tubulin to destabilize microtubule
Nat Commun 2016 Jun 30;7:12103.PMID:27357539DOI:10.1038/ncomms12103.
Molecules that alter the normal dynamics of microtubule assembly and disassembly include many anticancer drugs in clinical use. So far all such therapeutics target β-tubulin, and structural biology has explained the basis of their action and permitted design of new drugs. However, by shifting the profile of β-tubulin isoforms, cancer cells become resistant to treatment. Compounds that bind to α-tubulin are less well characterized and unexploited. The natural product Pironetin is known to bind to α-tubulin and is a potent inhibitor of microtubule polymerization. Previous reports had identified that Pironetin reacts with lysine-352 residue however analogues designed on this model had much lower potency, which was difficult to explain, hindering further development. We report crystallographic and mass spectrometric data that reveal that Pironetin forms a covalent bond to cysteine-316 in α-tubulin via a Michael addition reaction. These data provide a basis for the rational design of α-tubulin targeting chemotherapeutics.
The anticancer natural product Pironetin selectively targets Lys352 of alpha-tubulin
Chem Biol 2004 Jun;11(6):799-806.PMID:15217613DOI:10.1016/j.chembiol.2004.03.028.
Pironetin is a potent inhibitor of tubulin assembly and arrests cell cycle progression in M phase. Analyses of its structure-activity relationships suggested that Pironetin covalently binds tubulin. To determine the binding site of Pironetin, we synthesized biotinylated Pironetin, which inhibited tubulin assembly both in vitro and in situ. The biotinylated Pironetin selectively and covalently bound with tubulin. Partial digestion of biotinylated pironetin-treated tubulin by several proteases revealed that the binding site is the C-terminal portion of alpha-tubulin. By systematic alanine scanning, the Pironetin binding site was determined to be Lys352 of alpha-tubulin. Lys352 is located at the entrance of a small pocket of alpha-tubulin, and this pocket faces the beta-tubulin of the next dimer. This is the first compound that covalently binds to the alpha subunit of tubulin and Lys352 of alpha-tubulin and inhibits the interaction of tubulin heterodimers.