Rottlerin
(Synonyms: 粗糠柴苦素,Mallotoxin; NSC 56346; NSC 94525) 目录号 : GC10820
Rottlerin 是从Mallotus Philippinensis中纯化得到的天然产物，是一种特异性蛋白激酶C（PKC）抑制剂，对PKCδ的 IC50 值为3-6μM，对PKCα、β、γ 的IC50值为30-42μM，对PKCε、η、ζ 的IC50值为 80-100μM。
Cas No.:82-08-6
Sample solution is provided at 25 µL, 10mM.
Rottlerin, a natural product purified from Mallotus Philippinensis, is a specific PKC inhibitor, with IC50 values for PKCδ of 3-6μM, PKCα, β, γ of 30-42μM, PKCε, η, ζ of 80-100μM[1]. Rottlerin also acts as a direct mitochondrial uncoupler, and stimulates autophagy by targeting a signaling cascade upstream of mTORC1[2]. Rottlerin is often used in the research of viral infections, neurodegenerative diseases and metabolic disorders[3][4].
In vitro, treatment of human microvascular endothelial cells (HMVECs) with Rottlerin (5 or 20μM; 2, 6, 24h) significantly inhibits the proliferation of cells by reducing thymidine incorporation by 75%–80%, dose-dependently inhibits tube formation on Matrigel with complete inhibition at 20μM after 18h, and reduces cyclin D-1 mRNA to 50%, 40%, and 20% of control levels at 2, 6, and 24h, respectively, with a similar trend observed for protein levels[5]. Additionally, Rottlerin exhibits significant antiproliferative effects on various cell lines, with IC50 values of 12μM for C6 cells (24h), 5μM for C6 cells (48h), 7μM for T98G cells (48h), and 9μM for U138MG cells (48h); reduces the number of S-phase cells in C6, T98G, and U138MG cells at 5μM (24h); and induces DNA laddering in HL-60 cells at 5μM (24h)[6].
In vivo, Rottlerin (20mg/kg/day; p.o.; 6 weeks) significantly inhibited the growth of AsPC-1 pancreatic cancer xenografts in Balb/c nude mice, decreased the expression of cell proliferation markers (Ki67 and PCNA) in tumor tissues, increased the activation of apoptosis markers (caspase-3 and PARP), and suppressed the expression of proteins related to angiogenesis (VEGF, VEGFR, Cox-2, IL-8) and epithelial–mesenchymal transition (Slug, Snail, MMP-2, MMP-9), while upregulating the expression of E-cadherin[7].
References:
[1] Gschwendt M, Müller HJ, Kielbassa K, et al. Rottlerin, a novel protein kinase inhibitor. Biochem Biophys Res Commun. 1994;199(1):93-98.
[2] Balgi AD, Fonseca BD, Donohue E, et al. Screen for chemical modulators of autophagy reveals novel therapeutic inhibitors of mTORC1 signaling. PLoS One. 2009;4(9):e7124.
[3] Lama Z, Gaudin Y, Blondel D, Lagaudrière-Gesbert C. Kinase inhibitors tyrphostin 9 and rottlerin block early steps of rabies virus cycle. Antiviral Res. 2019;168:51-60.
[4] Zhang D, Anantharam V, Kanthasamy A, Kanthasamy AG. Neuroprotective effect of protein kinase C delta inhibitor rottlerin in cell culture and animal models of Parkinson's disease. J Pharmacol Exp Ther. 2007;322(3):913-922.
[5] Valacchi G, Pecorelli A, Sticozzi C, et al. Rottlerin exhibits antiangiogenic effects in vitro. Chem Biol Drug Des. 2011;77(6):460-470.
[6] Parmer TG, Ward MD, Hait WN. Effects of rottlerin, an inhibitor of calmodulin-dependent protein kinase III, on cellular proliferation, viability, and cell cycle distribution in malignant glioma cells. Cell Growth Differ. 1997;8(3):327-334.
[7] Huang M, Tang SN, Upadhyay G, et al. Rottlerin suppresses growth of human pancreatic tumors in nude mice, and pancreatic cancer cells isolated from Kras(G12D) mice. Cancer Lett. 2014;353(1):32-40.
Rottlerin 是从Mallotus Philippinensis中纯化得到的天然产物，是一种特异性蛋白激酶C（PKC）抑制剂，对PKCδ的 IC50 值为3-6μM，对PKCα、β、γ 的IC50值为30-42μM，对PKCε、η、ζ 的IC50值为 80-100μM[1]。Rottlerin还作为一种直接的线粒体解偶联剂，通过靶向mTORC1上游的信号级联来刺激自噬[2]。Rottlerin常用于病毒感染、神经退行性疾病和代谢紊乱的研究[3][4]。
在体外，用Rottlerin（5或20μM；2、6、24小时）处理人微血管内皮细胞（HMVEC）显著抑制细胞增殖，使胸腺嘧啶掺入减少75%–80%，呈剂量依赖性地抑制Matrigel上的管状结构形成，20μM时在18小时后完全抑制，并在2、6和24小时时分别使cyclin D-1 mRNA 降低至对照组的 50%、40%和20%，蛋白水平也呈现类似趋势[5]。此外，Rottlerin对多种细胞系表现出显著的抗增殖效果，C6细胞（24小时）的 IC50 值为 12μM，C6细胞（48 小时）的IC50值为5μM，T98G细胞（48 小时）的IC50值为7μM，U138MG细胞（48小时）的IC50值为9μM；在5μM（24小时）时减少C6、T98G和U138MG细胞中S期细胞的数量并诱导HL-60细胞中DNA梯状条带的形成[6]。
在体内，Rottlerin（20mg/kg/天；口服；6周）显著抑制了Balb/c裸鼠中AsPC-1胰腺癌异种移植瘤的生长，降低了肿瘤组织中细胞增殖标志物（Ki67和PCNA）的表达，增加了凋亡标志物（caspase-3 和 PARP）的激活，并抑制了与血管生成（VEGF、VEGFR、Cox-2、IL-8）和上皮 - 间充质转化（Slug、Snail、MMP-2、MMP-9）相关的蛋白表达，同时上调了E-cadherin的表达[7]。
| Cell experiment [1]: | |
Cell lines | HMVEC cells |
Preparation Method | Primary HMVEC cells were cultured in EBM-2 medium supplemented with EGM-2 Single Quots. All experiments were performed on near-confluent cultures from passages 3–9 made quiescent by incubation for 18–24h in serumfree EBM-2 medium. Cells were treated with 5 or 20μM Rottlerin for different times(2, 6, 24h) and then with TNFa (10ng⁄mL) for 4h. After treatment, cells were lysed in RIPA buffer (50mM Hepes–KOH, pH 7.4, 250mM NaCl, 1% NP-40, 5mM DTT, protease inhibitors). Total protein concentrations were determined using Bio-Rad protein assay reagent according to the manufacturer's instructions. Equal amounts of proteins (30μg) were separated by SDS–PAGE using NOVEX 4–20% Tris-Glycine Pre-Cast Gel and transferred to a nitrocellulose membrane. The membranes were blocked for 1h with 5% non-fat dry milk in PBS, 0.1% Tween 20 and incubated with the primary antibodies for cyclin D-1, ECE-1, and β-actin. Then, the membranes were incubated with the appropriate secondary antibodies, and after washing, bound antibodies were visualized by enhanced chemiluminescence. Chemiluminescence was captured on X-ray film, and resulting band densities were scanned and quantified using NIH IMAGE software. |
Reaction Conditions | 5 or 20μM; 2, 6, 24h |
Applications | Rottlerin reduces cyclin D-1 to 50%, 40%, and 20% of control levels at 2, 6, and 24h, respectively. |
| Animal experiment [2]: | |
Animal models | Balb/c nude mice |
Preparation Method | AsPC-1 cells (2×106 cells mixed with Matrigel, 50:50 ratio) were injected subcutaneously into the flanks of Balb/c nu/nu mice (4–6 weeks old, n=7). Rottlerin (0 or 20mg/kg) was administered to tumor-bearing mice through gavage (5 days per week, once daily) for 6 weeks. At the end of the experiment, all the mice from control and rottlerin-treated groups (seven mice in each group) were euthanized. Tumors from each mouse were isolated and divided into two groups, the first group for the Western blot analysis and the second group for immunohistochemistry. |
Dosage form | 20mg/kg/day for 6 weeks; p.o. |
Applications | Rottlerin significantly inhibited the growth of AsPC-1 pancreatic cancer xenografts in Balb/c nude mice, decreased the expression of cell proliferation markers (Ki67 and PCNA) in tumor tissues, increased the activation of apoptosis markers (caspase-3 and PARP), and suppressed the expression of proteins related to angiogenesis (VEGF, VEGFR, Cox-2, IL-8) and epithelial–mesenchymal transition (Slug, Snail, MMP-2, MMP-9), while upregulating the expression of E-cadherin. |
References: | |
| Cas No. | 82-08-6 | SDF | |
| 别名 | 粗糠柴苦素,Mallotoxin; NSC 56346; NSC 94525 | ||
| 化学名 | (Z)-1-(6-(3-acetyl-2,4,6-trihydroxy-5-methylbenzyl)-5,7-dihydroxy-2,2-dimethyl-2H-chromen-8-yl)-3-phenylprop-2-en-1-one | ||
| Canonical SMILES | OC1=C2C(OC(C)(C)C=C2)=C(C(/C=C\C3=CC=CC=C3)=O)C(O)=C1CC(C(O)=C(C)C(O)=C4C(C)=O)=C4O | ||
| 分子式 | C30H28O8 | 分子量 | 516.55 |
| 溶解度 | >12.5mg/mL in DMSO (ultrasonic and warming and heat to 60°C) | 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。 | ||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.9359 mL | 9.6796 mL | 19.3592 mL |
| 5 mM | 0.3872 mL | 1.9359 mL | 3.8718 mL |
| 10 mM | 0.1936 mL | 0.968 mL | 1.9359 mL |
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| 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 | ||||||||||
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2.
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