HET0016
(Synonyms: N-hydroxy-N'-(4-n-butyl-2-methylphenyl)Formamidine) 目录号 : GC11540
HET0016 是一种高选择性的20-羟基花生四烯酸（20-HETE）合成酶抑制剂，对重组CYP4A1、CYP4A2和CYP4A3催化20-HETE合成作用的IC50值分别为17.7nM, 12.1nM和20.6nM。
Cas No.:339068-25-6
Sample solution is provided at 25 µL, 10mM.
HET0016 is a highly selective inhibitor of 20-hydroxy eicosatrienoic acid (20-HETE) synthase. The IC50 values for the catalytic synthesis of 20-HETE by recombinant CYP4A1, CYP4A2, and CYP4A3 are 17.7nM, 12.1nM, and 20.6nM, respectively [1-2]. 20-HETE is an effective vasoconstrictor that can inhibit Na+ transport in the proximal tubule and thick ascending limb of the loop of Henle (TALH) [3]. HET0016 can be used to inhibit angiogenesis and tumor growth [4].
In vitro, treatment with HET0016 (100μM; 24, 48h) significantly reduced the motility of 4T1 and MDA-MB-231 cells, the total area of cell invasion decreased, and cell migration also showed a similar reduction [4]. Treatment with HET0016 (1, 10μM; 48h) dose-dependently inhibited the cell proliferation of the 9L gliosarcoma cell line. HET0016 could inhibit proliferation induced by epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) and reduce the phosphorylation of PDGF receptors [5].
In vivo, HET0016 (10mg/kg/day; 5d; i.p.) Treatment reduced tumor growth in breast cancer xenograft model mice, and decreased tumor volume in both the early treatment group and the delayed treatment group [6]. HET0016 (10mg/kg/day; once every 12 hours for three days; i.p.) treatment significantly reduced the brain lesion volume and neurological deficits in collagenase-induced Intracerebral Hemorrhage (ICH) model mice, and decreased neuronal death, ROS production, gelatin dissolution activity and inflammatory response 3 days after ICH [7].
References:
[1] Seki T, et al. Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N'-(4-butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis. Biol Pharm Bull. 2005 Sep;28(9):1651-4.
[2] Borin T F, Zuccari D A P C, Jardim-Perassi B V, et al. HET0016, a selective inhibitor of 20-HETE synthesis, decreases pro-angiogenic factors and inhibits growth of triple negative breast cancer in mice[J]. PLoS One, 2014, 9(12): e116247.
[3] Hoagland K M, Flasch A K, Roman R J. Inhibitors of 20-HETE formation promote salt-sensitive hypertension in rats[J]. Hypertension, 2003, 42(4): 669-673.
[4] Borin TF, et al. HET0016 decreases lung metastasis from breast cancer in immune-competent mouse model. PLoS One. 2017 Jun 13;12(6): e0178830.
[5] Guo M, Roman R J, Fenstermacher J D, et al. 9L gliosarcoma cell proliferation and tumor growth in rats are suppressed by N-hydroxy-N′-(4-butyl-2-methylphenol) formamidine (HET0016), a selective inhibitor of CYP4A[J]. The Journal of pharmacology and experimental therapeutics, 2006, 317(1): 97-108.
[6] Borin T F, Zuccari D A P C, Jardim-Perassi B V, et al. HET0016, a selective inhibitor of 20-HETE synthesis, decreases pro-angiogenic factors and inhibits growth of triple negative breast cancer in mice[J]. PLoS One, 2014, 9(12): e116247.
[7] Han X, Zhao X, Lan X, et al. 20-HETE synthesis inhibition promotes cerebral protection after intracerebral hemorrhage without inhibiting angiogenesis. J Cereb Blood Flow Metab. 2019;39(8):1531-1543.
HET0016 是一种高选择性的20-羟基花生四烯酸（20-HETE）合成酶抑制剂，对重组CYP4A1、CYP4A2和CYP4A3催化20-HETE合成作用的IC50值分别为17.7nM, 12.1nM和20.6nM [1-2]。20-HETE是一种有效的血管收缩剂，能够抑制近端小管和亨利氏袢升支粗管(TALH)中的Na+转运[3]。HET0016可用于抑制血管生成和肿瘤生长 [4]。
在体外，HET0016（100μM; 24,48h）处理显著降低了4T1和MDA-MB-231细胞的运动性，细胞侵袭的总面积减少，细胞迁移也显示出类似的降低 [4]。HET0016（1,10μM; 48h）处理剂量依赖性地抑制了9L胶质肉瘤细胞系的细胞增殖。HET0016能够抑制表皮生长因子（EGF）和血小板衍生生长因子（PDGF）诱导的增殖并减少PDGF受体的磷酸化 [5]。
在体内，HET0016（10mg/kg/day; 5d; i.p.）治疗减少了乳腺癌异种移植模型小鼠的肿瘤生长，在早期治疗组和延迟治疗组中均降低了肿瘤体积 [6]。HET0016（10mg/kg/day; 每12h一次, 持续三天; i.p.）治疗显著减少了胶原酶诱导的脑出血（ICH）模型小鼠脑病变体积和神经功能缺损，并减少了ICH后3天的神经元死亡、ROS 产生、明胶溶解活性和炎症反应 [7]。
| Cell experiment [1]: | |
Cell lines | 4T1 luciferase positive cells and MDA-MB-231 cells |
Preparation Method | Wound healing assay was performed to detect the potential of HET0016 treatment to decrease migration malignant cells. 4T1 luciferase positive cells and MDA-MB-231 cells achieving 80-90% of confluency in 6 well plates were starved overnight with 0.5% FBS for cell cycle synchronization. Then, cells were treated with 100μM of HET0016 for 24 and 48 hours in 2% FBS media, and microphotographed every 24 hours. The wound size was measured using Image J software (NIH) by drawing a rectangular region of interest to quantify the visible area of wound. |
Reaction Conditions | 100μM; 24 and 48h |
Applications | HET0016 significantly reduced the motility of 4T1 luciferase positive cells and MDA-MB-231 cells, the total area of cell invasion decreased, and cell migration also showed a similar reduction. |
| Animal experiment [2]: | |
Animal models | Athymic nude mice |
Preparation Method | Athymic nude female mice (n = 28) 6–8 week-old and weighing 20–25 grams obtained from Charles River laboratory (Frederick, MD) were used in all experiments. After tumor implantation the animals received treatment with HET0016 (10mg/kg per day) or phosphate buffered saline (PBS) containing 10% of dimethyl sulfoxide (DMSO) and 10% cremophor as a vehicle control intraperitoneally (IP) for five days during the week (Monday through Friday). To better evaluate the effect of treatment, the animals were divided into four treatment groups receiving early treatment (beginning on the day of tumor inoculation) or delayed (on day 8 after inoculation of the tumor) and maintained for 21 or 28 days. Group 1 (n = 6) starting treatment early on day 0 until day 21; Group 2 (n = 4) also starting treatment early on day 0, however, was maintained for 28 days; Group 3 (n = 6) starting treatment delayed, after tumor establishment on day 8 until day 21 and Group 4 (n = 4) also starting treatment delayed on day 8, however, was maintained for 28 days. Eight animals were used as control receiving vehicle, four were maintained for 21 days and others four were maintained for 28 days. |
Dosage form | 10mg/kg/day for 5 days; i.p. |
Applications | HET0016 treatment decreased tumor volume in all animals in both early and delayed treatment groups. |
References: | |
| Cas No. | 339068-25-6 | SDF | |
| 别名 | N-hydroxy-N'-(4-n-butyl-2-methylphenyl)Formamidine | ||
| 化学名 | N-(4-butyl-2-methylphenyl)-N'-hydroxy-methanimidamide | ||
| Canonical SMILES | CCCCC1=CC=C(/N=C/NO)C(C)=C1 | ||
| 分子式 | C12H18N2O | 分子量 | 206.3 |
| 溶解度 | ≤14mg/ml in ethanol;14mg/ml in DMSO;14mg/ml in dimethyl formamide | 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 | 4.8473 mL | 24.2365 mL | 48.4731 mL |
| 5 mM | 0.9695 mL | 4.8473 mL | 9.6946 mL |
| 10 mM | 0.4847 mL | 2.4237 mL | 4.8473 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 |
| 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，
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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Quality Control & SDS
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- Purity: >98.50%
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