NAT10 (N-acetyltransferase 10)

当暴露于重塑蛋白 (10–40 μM) 1–7 天时，AR 阳性和 AR 阴性前列腺癌细胞均表现出对 NAT10 功能和细胞增殖的剂量依赖性抑制 [2]。在前列腺癌细胞中，remodelin（20 μM，24 小时）抑制 NAT10 并减慢 DNA 复制 [2]。在 LmnaG609G/G609G 成纤维细胞中，remodelin（1 μM，7 天）可减少核形态异常并促进基因组稳定性 [3]。

在肿瘤异种移植裸鼠模型中，remodelin（2 或 20 mg/kg，腹腔注射，每两天一次，持续 4 周）可显着减缓 AR 阴性前列腺癌的生长 [2]。 Remodelin（100 mg/kg，口服）可显着延长早衰综合征 (HGPS) LmnaG609G/G609G 小鼠模型的健康寿命并抑制 NAT10 [3]。在小鼠中，Remodelin（5 mg/kg，口服）的口服生物利用度 (F%) 为 43.5%，T1/2 为 1.81 小时 [3]。小鼠重塑蛋白药代动力学参数：途径剂量 (mg/kg)、T1/2 (h)、Tmax (h)、Cmax (ng/mL)、AUC0-t (ng/h/mL)、AUC0-Ɲ (ng/h) /mL), MRT_last (h), F(%) po 5 1.81 0.25 409 235 259 0.84 43.5%

赖氨酸乙酰转移酶（KAT）测定。[1]
使用荧光HAT测定试剂盒进行KAT测定，使用从HEK293细胞纯化的NAT10和5μg纯化的富含MAP的猪Tubulin作为底物。重塑和可点击分子2在50μM下使用。[1]

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圆二色光谱法。[1]
CD实验使用Chirascan圆二色分光光度计进行。将200μl最终浓度为10μM的纯化FLAG-NAT10（TBS+0.1%NP-40）置于光路长度为1mm的石英试管中，转移至分光光度计。CD扫描在25°C下记录，波长范围为180至350 nm，响应时间为1s，间距为1 nm，带宽为1.5 nm。加入溶解在DMSO中的化合物1，并在记录扫描之前将溶液温育5分钟。缓冲液减去CD光谱，在300 nm处校正零并归一化（摩尔椭圆度θ以105°cm2 dmol−1为单位）。

细胞增殖测定[2]
细胞类型：前列腺癌细胞系 VCaP、PC3 和 DU145
测试浓度： 0、10、20、40 μM
孵育时间：1、2、7天
实验结果：抑制NAT10并抑制AR阳性和AR阴性前列腺癌细胞的生长。与对照组相比，随着时间的推移，细胞增殖活性显着降低。以剂量依赖性方式降低集落形成能力。

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免疫荧光[2]
细胞类型：前列腺癌细胞系 VCaP、PC3 和 DU145
测试浓度： 0,10， 20,40 μM
孵育时间：24小时
实验结果：与相比，阳性标记率和荧光强度均显着下降到对照组。与对照组相比，IdU 的染色焦点和 CldU 的染色焦点均显着减少。

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蛋白质印迹分析[3]
细胞类型： 来自 LmnaG609G/G609G 和野生型 (WT) 同窝小鼠的皮肤成纤维细胞
测试浓度： 1 μM
孵育时间： 7 天
实验结果： 降低了 DNA dou 的较高水平

Animal/Disease Models: PC-3 cells tumor xenograft model in nude athymic BALB/c nu/nu (nude) mice[2]
Doses: 2 or 20 mg/ kg
Route of Administration: intraperitoneal (ip)injection, once every two days for 4 weeks
Experimental Results: Dramatically decreased AR-negative prostate cancer tumor growth. In the high-dose group, xenograft tumor weight at the endpoint was much smaller than that in the low- dose group.

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Animal/Disease Models: LmnaG609G/G609G hutchinson-gilford progeria syndrome (HGPS) mouse model[3]
Doses: 100 mg/kg
Route of Administration: po (oral gavage), daily schedule for 3 weeks of age onward, until the end-point Ameliorated age-dependent weight loss.
Experimental Results: Ameliorated cardiac pathology. Led to the dramatic amelioration of HGPS cardiac pathologies, including reduction of adventitial fibrosis of the aorta, rescue of vascular smooth muscle cell loss, and salvage of smooth muscle actin (SMA) loss , both in the aorta and the coronary arteries.

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Animal/Disease Models: WT Mice (pharmacokinetic/PK assay)[3]
Doses: 5 mg/kg
Route of Administration: Oral gav

[1]. Larrieu D, et al. Chemical inhibition of NAT10 corrects defects of laminopathic cells. Science. 2014 May 2;344(6183):527-32.
[2]. Ma N, et.al. Inhibition of N-Acetyltransferase 10 Suppresses the Progression of Prostate Cancer through Regulation of DNA Replication. Int J Mol Sci. 2022 Jun 12;23(12):6573.
[3]. Balmus G, et.al. Targeting of NAT10 enhances healthspan in a mouse model of human accelerated aging syndrome. Nat Commun. 2018 Apr 27;9(1):1700.
[4]. Zhang X, et.al. N-Acetyltransferase 10 Enhances Doxorubicin Resistance in Human Hepatocellular Carcinoma Cell Lines by Promoting the Epithelial-to-Mesenchymal Transition. Oxid Med Cell Longev. 2019 Jul 1;2019:7561879.

C15H14N4S

282.36

282.09391

C, 63.81; H, 5.00; N, 19.84; S, 11.35

949912-58-7

Remodelin hydrobromide;1622921-15-6

Off-white to light yellow solid

3.3

89.3Ų

N#CC1=CC=C(C2=CSC(N/N=C3CCCC/3)=N2)C=C1

XAEJIFARBQJLML-UHFFFAOYSA-N

InChI=1S/C15H14N4S/c16-9-11-5-7-12(8-6-11)14-10-20-15(17-14)19-18-13-3-1-2-4-13/h5-8,10H,1-4H2,(H,17,19)

4-[2-(2-cyclopentylidenehydrazinyl)-4-thiazolyl]-benzonitrile

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。