ML-323
目录号 : GC17635
ML-323是一种可逆性强、高效的USP1-UAF1的选择性抑制剂，IC50值为76nM。ML-323对游离酶的Ki值为68nM。
Cas No.:1572414-83-5
Sample solution is provided at 25 µL, 10mM.
_1-3.$$$39978408@51.jpg');)
_1-9.$$$38538795@65.jpg');)
_1-9.$$$39112701@51.jpg');)
_1-7.$$$37316672@70.jpg');)
_366-372.$$$36198801@70.jpg');)
.$$$38565142@65.jpg');)
_1867-1883.$$$33248023@67.jpg');)
_eads6055.$$$39977546@45.jpg');)
_eadm9805.$$$40080571@45.jpg');)
_eadj3020.$$$39666821@45.jpg');)
_1-20.$$$39757301@28.jpg');)
_1-24.$$$38898113@28.jpg');)
_273-287.$$$36806353@46.jpg');)
_1-16.$$$37156877@44.jpg');)
_1-19.$$$37460805@44.jpg');)
_1291-1307.$$$34518654@46.jpg');)
ML-323 is a highly reversible and highly effective selective inhibitor of USP1-UAF1 with an IC50 value of 76nM. The Ki value of ML-323 for free enzymes is 68nM. USP1-UAF1 is closely related to the DNA damage response and can be used for the study of anti-tumor drug resistance [1]. ML-323 has anti-cancer activity against various cancers [2-3].
In vitro, in the Ub-Rho experiment, ML-323 (0.08-114μM; 1-2h) exhibited dose-dependent inhibition of USP1-UAF1 with an IC50 value of 76nM [1]. ML-323 (0-200μM; 48h) inhibited the growth of HCCLM3 and SMMC-7721 HCC cells and the formation of colonies in a dose-dependent manner [4]. ML-323 (0-400nM; 0, 4, 8 and 16h) could down-regulate the expression of USP1 in colorectal cancer cells in a dose-dependent and time-dependent manner [5].
In vivo, the injection treatment of ML-323 (10mg/kg; 7 days) significantly increased the body weight of mice with colitis models and decreased the MPO activity in colonic tissues and ulcer areas, and had a significant inhibitory effect on the levels of inflammatory markers (TNF-α, IL-6, IL-17 and IL-1β) in the colonic tissues of colitis mice [6]. ML-323 (25 and 50mg/kg/day; oral; twice a week for 8 weeks) treatment significantly reduced the body weight of mice fed a high-fat diet (HFD), improved insulin and glucose sensitivity, and significantly decreased the fat content and size of adipocytes in white adipose tissue, reduced lipid accumulation, triglycerides, free fatty acids and macrophage infiltration in the liver of the mice [7].
References:
[1] Liang Q, Dexheimer T S, Zhang P, et al. A selective USP1–UAF1 inhibitor links deubiquitination to DNA damage responses[J]. Nature chemical biology, 2014, 10(4): 298-304.
[2] Sun Y, Sha B, Huang W, et al. ML323, a USP1 inhibitor triggers cell cycle arrest, apoptosis and autophagy in esophageal squamous cell carcinoma cells[J]. Apoptosis, 2022, 27(7): 545-560.
[3] Song B, Jiang Y, Jiang Y, et al. ML323 suppresses the progression of ovarian cancer via regulating USP1-mediated cell cycle[J]. Frontiers in Genetics, 2022, 13: 917481.
[4] Wang L, Hu T, Shen Z, et al. Inhibition of USP1 activates ER stress through Ubi-protein aggregation to induce autophagy and apoptosis in HCC[J]. Cell Death & Disease, 2022, 13(11): 951.
[5] Xu X, Mei X, Han K, et al. The deubiquitinating enzyme USP1 is auto-ubiquitinated and destabilized by ML323 in colorectal cancer cells[J]. Eurasian J Med Oncol, 2023, 7: 174-179.
[6] Lai Y, Liu J, Hu X, et al. N6-methyladenosine (m6A)-forming enzyme METTL3 controls UAF1 stability to promote inflammation in a model of colitis by stimulating NLRP3[J]. Scientific Reports, 2025, 15(1): 5876.
[7] Kim M S, Baek J H, Lee J A, et al. Deubiquitinase USP1 enhances CCAAT/enhancer-binding protein beta (C/EBPβ) stability and accelerates adipogenesis and lipid accumulation[J]. Cell Death & Disease, 2023, 14(11): 776.
ML-323是一种可逆性强、高效的USP1-UAF1的选择性抑制剂，IC50值为76nM。ML-323对游离酶的Ki值为68nM。USP1-UAF1与DNA损伤反应密切相关，可用于抗肿瘤耐药性的研究 [1]。ML-323对多种癌症具有抗癌活性 [2-3]。
在体外，在Ub-Rho实验中，ML-323（0.08-114μM; 1-2h）对USP1-UAF1剂量依赖性抑制，其IC50值为76nM [1]。ML-323（0-200μM; 48h）以剂量依赖性方式抑制HCCLM3和SMMC-7721 HCC细胞的生长和菌落形成 [4]。ML-323（0-400nM; 0, 4, 8和16h）能够以剂量依赖性和时间依赖性方式下调结直肠癌细胞中USP1的表达 [5]。
在体内，ML-323（10mg/kg; 7d）的注射治疗显著增加了结肠炎模型小鼠的体重并降低结肠组织和溃疡区域的MPO活性，对结肠炎小鼠结肠组织中炎症标志物（TNF-α、IL-6、IL-17和IL-1β）水平有显著抑制作用[6]。ML-323（25 and 50mg/kg/day; oral; twice a week, 8 weeks）治疗显著减轻高脂饮食（HFD）小鼠体重以及改善胰岛素和葡萄糖敏感性，并且显著降低白色脂肪组织中的脂肪量和脂肪细胞大小，减少了小鼠肝脏中的脂质积累、甘油三酯、游离脂肪酸和巨噬细胞浸润 [7]。
| Kinase experiment [1]: | |
Preparation Method | To determine the IC50 of ML323 in inhibiting USP1-UAF1, the inhibitor was added at seven different concentrations to the assay containing 150nM USP1-UAF1 and 3μM K63-linked diubiquitin or 300nM USP1-UAF1 and 3μM Ub-PCNA in a buffer containing 50mM HEPES (pH 7.8), 0.1mg ml−1 BSA, 0.5mM EDTA and 1mM or 5mM DTT for 1–2 h at 37°C. The reaction was quenched by the addition of Laemmli sample buffer. In the in vitro ML-323 selectivity test, six different concentrations (0.08–114μM) of ML-323 and 3μM K63-linked diubiquitin were incubated individually with 7.5nM USP7, 30nM USP2, 15nM USP5, 255nM USP8, 100nM USP11, 600nM USP21 and 600nM USP46-UAF1. |
Reaction Conditions | 0.08-114μM; 1-2h |
Applications | In the Ub-Rho assay, ML-323 exhibited dose-dependent inhibition of USP1-UAF1, with its IC50 value being 76nM. ML323 showed no significant inhibitory effect on other proteases and kinases. |
| Cell experiment [2]: | |
Cell lines | HCC cell lines |
Preparation Method | HCC cell lines HCCLM3, HepG2, Huh7, and SMMC-7721 were uniformly seeded in a 96-well plate (3 × 103 cells/well) and treated with ML-323 or DMSO (0.1%) for 48h. Cell viability was determined using a cell-counting-kit (CCK)-8 according to the manufacturer’s protocol. For the colony formation assay, 500 cells were seeded into a 6-well plate in triplicate, treated with DMSO or ML-323, and incubated for 10d. The colonies were treated with 4% paraformaldehyde and crystal violet, then counted. |
Reaction Conditions | 0-200μM; 48h |
Applications | ML-323 inhibited the growth of HCCLM3 and SMMC-7721 HCC cells in a dose-dependent manner, and also inhibited colony formation. |
| Animal experiment [3]: | |
Animal models | C57BL/6 mice (colitis models) |
Preparation Method | Male C57BL/6 mice (4-5 weeks, 17–19g) were purchased from Animal Experimental Center of Guangdong Medical University. Colitis was induced by administering 2.0% dextran sulfate sodium (DSS) in the drinking water for 7 days. Mice were provided with a standard diet and water ad libitum. UAF1 inhibitor (10mg/kg/day for 7 days of ML-323) was injected. Anesthesia was induced with 50mg/kg pentobarbital sodium, and mice were euthanized by cervical dislocation. Colon tissues, RAW264.7 macrophages, and serum samples were extracted. |
Dosage form | 10mg/kg/day for 7 days; inject |
Applications | Treatment with ML-323 significantly increased the body weight of mice and reduced the MPO activity in colonic tissues and ulcer areas. It also had a significant inhibitory effect on the levels of inflammatory markers (TNF-α, IL-6, IL-17 and IL-1β) in the colonic tissues of colitis mice. |
References: | |
| Cas No. | 1572414-83-5 | SDF | |
| 化学名 | (E)-1-(4-(1H-1,2,3-triazol-1-yl)phenyl)-N-(2-(2-isopropylphenyl)-5-methylpyrimidin-4(3H)-ylidene)methanamine | ||
| Canonical SMILES | CC(C1=CC=CC=C1C2=NC=C(/C(N2)=N\CC3=CC=C(N4C=CN=N4)C=C3)C)C | ||
| 分子式 | C23H24N6 | 分子量 | 384.48 |
| 溶解度 | ≥ 15.05mg/mL in DMSO | 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。 |
||
| 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 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.6009 mL | 13.0046 mL | 26.0092 mL |
| 5 mM | 0.5202 mL | 2.6009 mL | 5.2018 mL |
| 10 mM | 0.2601 mL | 1.3005 mL | 2.6009 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。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet