SIRT2 (IC50 = 10 μM); SIRT1 (IC50 = 21 μM); SIRT3 (IC50 = 67 μM); HDAC8; MDM-2/p53
Tenovin-6 Hydrochloride is more toxic to yeast than the less water-soluble tenovin-1, with an IC50 of 30 M for inhibiting the growth of S. cerevisiae cultures. In MCF-7 cells, endogenous K382-Ac p53 levels are quickly elevated by tenovin-6 hydrochloride[1].
Tenovin-6 Hydrochloride (0 to 15 μM) dose dependently increases the level of LC3-II in diverse cell types, and the increase is ATG5/7 dependent. In addition, the use of tenovin-6 hydrochloride prevents SQSTM1/p62 degradation caused by torin 1 and boosts the quantity and intensity of autophagic vesicles both with and without torin 1. The fusion between autophagosomes and lysosomes is unaffected by tenovin-6 hydrochloride, but it affects the acidification of autolysosomes and hinders lysosome hydrolytic activity. Tenovin-6 Hydrochloride inhibits autophagy, but this is unrelated to p53 activation, and inhibiting SIRT1/2 either through knockdown or knockout cannot mimic Tenovin-6 Hydrochloride's effect on LC3B accumulation[3].
Tenovin-6 Hydrochloride (0, 1, 2.5, 5 or 10 μM) potently inhibits cell proliferation in a dose- and time-dependent manner in all OCI-Ly1, DHL-10, U2932, RIVA, HBL1 and OCI-Ly10 cell lines. Tenovin-6 Hydrochloride consistently raises the level of LC3B-II in DLBCL cell lines by blocking the traditional autophagy pathway without turning on p53, and the rise is unrelated to SIRT1/2/3 or p53. Through the extrinsic cell-death pathway, tenovin-6 hydrochloride causes apoptosis[4].
Tenovin-6 Hydrochloride inhibits UM cell growth with IC50 values of 12.8 μM, 11.0 μM, 14.58 μM and 9.62 μM for 92.1, Mel 270, Omm 1, and Omm 2.3 cells, respectively[5].

Tenovin-6 Hydrochloride 比水溶性较差的 tenovin-1 对酵母的毒性更大，抑制酿酒酵母培养物生长的 IC50 为 30 M。在 MCF-7 细胞中，tenovin-6 盐酸盐可快速升高内源性 K382-Ac p53 水平[1]。 Tenovin-6 Hydrochronide（0 至 15 μM）剂量依赖性地增加多种细胞类型中 LC3-II 的水平，并且这种增加与 ATG5/7 相关。此外，使用 tenovin-6 盐酸盐可以防止 torin 1 引起的 SQSTM1/p62 降解，并提高有或没有 torin 1 的自噬囊泡的数量和强度。自噬体和溶酶体之间的融合不受 tenovin-6 盐酸盐的影响，但它影响自溶酶体的酸化并阻碍溶酶体的水解活性。 Tenovin-6 Hydrochloride 会抑制自噬，但这与 p53 激活无关，并且通过敲除或敲除来抑制 SIRT1/2 不能模拟 Tenovin-6 Hydrochronide 对 LC3B 积累的影响[3]。 Tenovin-6 Hydrochloride (0、1、2.5、5 或 10 μM) 以剂量和时间依赖性方式有效抑制所有 OCI-Ly1、DHL-10、U2932、RIVA、HBL1 和 OCI-Ly10 细胞系的细胞增殖。 Tenovin-6 Hydrochloride 通过阻断传统的自噬途径而不开启 p53，持续提高 DLBCL 细胞系中 LC3B-II 的水平，并且这种升高与 SIRT1/2/3 或 p53 无关。 tenovin-6盐酸盐通过外源性细胞死亡途径引起细胞凋亡[4]。 Tenovin-6 Hydrochronide 抑制 UM 细胞生长，对于 92.1、Mel 270、Omm 1 和 Omm 2.3 细胞的 IC50 值分别为 12.8 μM、11.0 μM、14.58 μM 和 9.62 μM[5]。

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在MCF-7细胞中，Tenovin-6 处理6小时后可浓度依赖性地增加p53蛋白水平，其效果略优于tenovin-1。[1]
在转染了p53表达载体的H1299细胞中，即使总p53水平保持不变，Tenovin-6 也能提高p53在赖氨酸382位点的乙酰化水平。[1]
在H1299细胞中，Tenovin-6 能增加α-微管蛋白在赖氨酸40位点的乙酰化水平，此效应可被SirT2过表达所减弱。[1]
在72小时细胞活性实验中，Tenovin-6 对ARN8黑色素瘤细胞的毒性高于tenovin-1。[1]
用Tenovin-6 处理ARN8细胞2小时后，更换新鲜培养基继续培养4天，可抑制细胞生长。[1]
它抑制酿酒酵母生长的IC50为30 µM。[1]
它对一组51种纯化激酶的活性、爪蟾卵提取物中的DNA复制实验以及人细胞提取物中的RNA聚合酶I转录实验均无显著影响。[1]

Tenovin-6 Hydrochloride (50 mg/kg, ip) 可抑制小鼠肿瘤的生长[1]。

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每日腹腔注射Tenovin-6（50 mg/kg）可显著延缓SCID小鼠中ARN8黑色素瘤细胞来源的异种移植瘤在15天内的生长。[1]

Fluor de Lys 荧光检测系统使用纯化的成分进行检测。有用的 FdL 底物的使用浓度为 7 M，NAD+ 的使用浓度为 1 mM。 Tenovin 可溶于 DMSO，反应中 DSMO 的最终浓度低于 0.25%。 SirT1 和 HDAC8 的每个反应使用 1 个单位的酶，SirT2 和 SirT3 的每个反应使用 5 个单位的酶。反应在37℃下进行一小时。

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使用纯化的人源SirT1、SirT2、SirT3或HDAC8酶进行体外去乙酰化实验。反应在荧光检测体系中进行，使用特定的乙酰化肽段底物和NAD+作为辅底物。化合物溶于DMSO后加入反应体系。在37°C孵育1小时后测定酶活。通过测试Tenovin-6 的递增浓度来确定IC50值。[1]
对Tenovin-6 抑制SirT1进行了Lineweaver-Burke分析，分别改变乙酰化肽底物或NAD+的浓度，以确定抑制模式。[1]

MTS 测定用于评估细胞活力。在 96 孔板中，将 UM 细胞接种到每个孔中（5,000 个细胞/孔），第二天用对照或 Tenovin-6（浓度从 0 增加到 20 M）处理 68 小时，然后以 20 L 加入 MTS /well 在 490 nm 波长下读取。 IC50通过曲线拟合S形剂量反应曲线来计算。

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为分析p53蛋白水平，用Tenovin-6 处理MCF-7细胞6小时，裂解后进行Western blot，使用p53抗体检测。用PCNA抗体作为上样对照。[1]
为分析乙酰化状态，将H1299细胞转染p53表达载体，用Tenovin-6 处理6小时，然后使用乙酰化p53（K382）抗体和总p53抗体进行Western blot分析。[1]
为检测乙酰化α-微管蛋白，将H1299细胞用Tenovin-6 处理16小时，同时加入曲古抑菌素A以抑制非sirtuin的HDACs，然后使用乙酰化α-微管蛋白（K40）抗体和总α-微管蛋白抗体进行Western blot分析。[1]
细胞活性通过吉姆萨染色评估，处理方法包括用Tenovin-6 持续处理72小时，或脉冲处理2小时后更换新鲜培养基再培养4天。[1]
通过BrdU标记和碘化丙啶染色结合流式细胞术分析细胞周期分布和细胞死亡。[1]

50 mg/kg
ARN8 melanoma cells were injected into the flank of SCID mice and allowed to develop into tumors.

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Female SCID mice were injected subcutaneously with 1 × 10^6 ARN8 melanoma cells suspended in Matrigel. Tumors were allowed to reach approximately 10 mm² in size. Tenovin-6 was dissolved in a vehicle containing 20% cyclodextrin and 10% DMSO. Treatment consisted of daily intraperitoneal injections of Tenovin-6 at 50 mg/kg. Control animals received the vehicle alone. Tumor dimensions were measured regularly with calipers, and volumes were calculated. Statistical significance was assessed using the Mann-Whitney U-test. [1]

Tenovin-6 is seven times more water soluble than tenovin-1. [1]
Its improved water solubility allowed for better pharmacokinetic evaluation compared to tenovin-1, but specific PK parameters (e.g., half-life, bioavailability) are not provided in the main text or supplemental data of this publication. [1]

Tenovin-6 shows primarily cytostatic effects on normal human dermal fibroblasts (NHDFs) with little cell death, in contrast to its cytotoxic effects on ARN8 tumor cells. [1]

[1]. Discovery, in vivo activity, and mechanism of action of a small-molecule p53 activator. Cancer Cell. 2008 May;13(5):454-63.

[2]. Exploitation of DHODH and p53 activation as therapeutic targets - a case study in polypharmacology [published online ahead of print, 2020 Sep 8]. J Biol Chem. 2020;jbc.RA119.012056.

[3]. Class III-specific HDAC inhibitor Tenovin-6 induces apoptosis, suppresses migration and eliminates cancer stem cells in uveal melanoma. Sci Rep. 2016 Mar 4;6:22622.

[4]. Tenovin-6 impairs autophagy by inhibiting autophagic flux. Cell Death Dis. 2017 Feb 9;8(2):e2608.

[5]. Tenovin-6 inhibits proliferation and survival of diffuse large B-cell lymphoma cells by blocking autophagy. Oncotarget. 2017 Feb 28;8(9):14912-14924.

Tenovin-6 was discovered through a forward chemical genetics screen for small molecules that activate a p53-dependent reporter in mammalian cells. [1]
It acts by inhibiting sirtuin deacetylases (SirT1/SirT2), leading to increased acetylation and stabilization of p53, as well as acetylation of other substrates like α-tubulin. [1]
Functional p53 contributes to but is not essential for its cytotoxic effects. [1]
Unlike DNA-damaging agents, Tenovin-6 does not activate the DNA damage response (no increase in phospho-Ser15 p53 or γH2AX). [1]
It represents a potential lead compound for cancer therapy and a tool for studying sirtuin biology. [1]

C₂₅H₃₅CLN₄O₂S

491.09

490.216

C, 61.14; H, 7.18; Cl, 7.22; N, 11.41; O, 6.52; S, 6.53

1011301-29-3

Tenovin-6;1011557-82-6

49871498

Yellow solid powder

106

4

4

9

33

616

0

Cl[H].S=C(N([H])C1C([H])=C([H])C(=C([H])C=1[H])N([H])C(C([H])([H])C([H])([H])C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])[H])=O)N([H])C(C1C([H])=C([H])C(=C([H])C=1[H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])=O

UBNCTIDXQDCEPI-UHFFFAOYSA-N

InChI=1S/C25H34N4O2S.ClH/c1-25(2,3)19-11-9-18(10-12-19)23(31)28-24(32)27-21-15-13-20(14-16-21)26-22(30)8-6-7-17-29(4)5;/h9-16H,6-8,17H2,1-5H3,(H,26,30)(H2,27,28,31,32);1H

4-tert-butyl-N-[[4-[5-(dimethylamino)pentanoylamino]phenyl]carbamothioyl]benzamide;hydrochloride

Tenovin-6 HCl; Tenovin6; Tnv-6; Tenovin 6; Tenovin-6

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)