ML323

Catalog No.S7529 Batch:S752902

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Technical Data

Formula

C23H24N6
Molecular Weight 384.48 CAS No. 1572414-83-5
Solubility (25°C)* In vitro DMSO 76 mg/mL (197.66 mM)
Ethanol 38 mg/mL (98.83 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description ML323 displays reversible, nanomolar inhibitory activity and excellent selectivity toward USP1/UAF1 with IC50 of 76 nM.
Targets
USP1-UAF1 [1]
(Cell-free assay)
76 nM
In vitro

ML323 inhibits the deubiquitination of PCNA and FANCD2 by inhibiting USP1–UAF1 activity in H596 cells. Moreover, ML323 potentiates cytotoxicity in NSCLC H596 cells and U2OS osteosarcoma cells by targeting two major DNA damage response pathways (TLS and FA). [1]
In vivo

ML323 is a selective USP1–UAF1 inhibitor that inhibit deubiquitinase activity of USP1–UAF1 complex.
Features USP1-UAF1-selective inhibitor.

Protocol (from reference)

Kinase Assay:

[1]
  • High-throughput screening

    For HTS, USP1-UAF1 activity is monitored using ubiquitin-rhodamine 110 as a substrate. The assay is miniaturized to a 4 μL volume in a 1,536-well format and is used to screen approximately 402,701 compounds in quantitative HTS mode, with each compound tested over a range of four to five concentrations. The assay shows robust performance with an average Z’factor of 0.8 throughout the screen.
Cell Assay:

[1]
  • Cell lines

    H596 cells

  • Concentrations

    ~30 μM

  • Incubation Time

    7-12 days

  • Method

    For the colony-forming assay, cells are seeded at a density of 300–500 cells per well in six-well plates and grown overnight. Cells are then treated with ML323 alone at the indicated concentrations. Cells treated with an equal volume of DMSO and saline are used as control. After 48 h of treatment, fresh growth medium is added, and cells are incubated for an additional 5-10 d to allow for colony formation. For UV combination treatment, the cells are treated with ML323 at the indicated concentrations or an equal volume of DMSO. After 48 h, the medium is removed, and cells are irradiated at 254 nm at the indicated dosage. Fresh growth medium is added, and the cells are incubated for an additional 5-10 d to allow for colony formation. The cells without UV irradiation but treated with ML323 or an equal volume of DMSO are used as controls and designated as 100%. After the formation of the colonies, cells are fixed with methanol and stained with 0.5% crystal violet. Colonies consisting of >50 cells are scored. The number of colonies is determined from triplicate plates. The dose-response curves are generated using GraphPad Prism and analyzed by using CalcuSyn to calculate the combination index.
Animal Study:

[2]
  • Animal Models

    Female C57BL/6J mice

  • Dosages

    10 mg/kg

  • Administration

    i.p.

Selleck's ML323 has been cited by 16 publications

Polyubiquitinated PCNA triggers SLX4-mediated break-induced replication in alternative lengthening of telomeres (ALT) cancer cells [ Nucleic Acids Res, 2024, gkae785] PubMed: 39291733
Multi-regulatory potency of USP1 on inflammasome components promotes pyroptosis in thyroid follicular cells and contributes to the progression of Hashimoto's thyroiditis [ Mol Med, 2024, 30(1):121] PubMed: 39134949
USP1 promotes the aerobic glycolysis and progression of T-cell acute lymphoblastic leukemia via PLK1/LDHA axis [ Blood Adv, 2023, 7(13):3099-3112] PubMed: 36912760
PCNA Ser46-Leu47 residues are crucial in preserving genomic integrity [ PLoS One, 2023, 18(5):e0285337] PubMed: 37205694
The Deubiquitinating Enzyme USP1 is Auto-Ubiquitinated and Destabilized by ML323 in Colorectal Cancer Cells [ Eurasian J Med Oncol, 2023, 7(2):174–179] PubMed: none
USP1-dependent RPS16 protein stability drives growth and metastasis of human hepatocellular carcinoma cells [ J Exp Clin Cancer Res, 2021, 40(1):201] PubMed: 34154657
A new role of GRP75-USP1-SIX1 protein complex in driving prostate cancer progression and castration resistance [ Oncogene, 2021, 40(25):4291-4306] PubMed: 34079090
Primaquine Diphosphate, a Known Antimalarial Drug, Blocks Vascular Leakage Acting Through Junction Stabilization [ Front Pharmacol, 2021, 12:695009] PubMed: 34149436
USP1-WDR48 deubiquitinase complex enhances TGF-β induced epithelial-mesenchymal transition of TNBC cells via stabilizing TAK1 [ Cell Cycle, 2021, 1-12] PubMed: 33461373
UAF1 deubiquitinase complexes facilitate NLRP3 inflammasome activation by promoting NLRP3 expression [ Nat Commun, 2020, 11(1):6042] PubMed: 33247121

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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