ML-323 |
Katalog-Nr.GC17635
One-Click Copy Product Info
|
ML-323 ist ein reversibler, potenter USP1-UAF1-Inhibitor mit einem IC50-Wert von 76 nM in einem Ub-Rho-Assay. Die gemessene Hemmkonstante von ML-323 fÜr das freie Enzym (Ki) betrÄgt 68 nM.
Products are for research use only. Not for human use. We do not sell to patients.
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.
| 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 | |
| Chemical Name | (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 | ||
| Formula | C23H24N6 | M.Wt | 384.48 |
| Löslichkeit | ≥ 15.05mg/mL in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.6009 mL | 13.0046 mL | 26.0092 mL |
| 5 mM | 520.2 μL | 2.6009 mL | 5.2018 mL |
| 10 mM | 260.1 μL | 1.3005 mL | 2.6009 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 18 reference(s) in Google Scholar.)