| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
IC50: 4.7 µM (UCK2)[1]
UCK2 Inhibitor-1 specifically targets uridine-cytidine kinase 2 (UCK2), an enzyme in the pyrimidine nucleoside salvage pathway. UCK2 is responsible for the ATP-dependent phosphorylation of uridine and cytidine to UMP and CMP. This salvage pathway is essential for maintaining nucleotide pools, especially in cells with high proliferation rates and in certain cancers where de novo pyrimidine synthesis is insufficient. UCK2 is also involved in the activation of nucleoside analog prodrugs, such as gemcitabine, decitabine, and azacitidine. By non-competitively inhibiting UCK2, the compound blocks the phosphorylation of these nucleosides, thereby inhibiting the synthesis of pyrimidine nucleotides and disrupting DNA/RNA synthesis. This leads to cytotoxic effects in tumor cells. The non-competitive mechanism indicates that UCK2 Inhibitor-1 binds to a site distinct from the uridine or ATP binding sites, possibly an allosteric site. |
|---|---|
| ln Vitro |
In vitro, UCK2 Inhibitor-1 is evaluated for its ability to inhibit UCK2 enzyme activity in a cell-free assay. The IC50 for UCK2 is 4.7 uM. The compound acts as a non-competitive inhibitor with respect to both uridine and ATP. It does not significantly inhibit the related enzyme UCK1 (IC50 >100 uM), demonstrating selectivity for UCK2 over UCK1. In cell-based assays, UCK2 Inhibitor-1 shows antiproliferative activity against cancer cell lines that rely on the salvage pathway for pyrimidine nucleotides. For example, in certain leukemia and lymphoma cell lines, the compound reduces cell viability with IC50 values in the low micromolar range (e.g., 5-20 uM). The compound also induces cytotoxic effects by disrupting nucleotide metabolism. In combination studies, UCK2 Inhibitor-1 can sensitize cancer cells to nucleoside analog chemotherapies (e.g., gemcitabine) by inhibiting their phosphorylation, thereby highlighting the role of UCK2 in drug activation. The compound is non-toxic to normal cells that rely primarily on de novo pyrimidine synthesis.
|
| Enzyme Assay |
A non-cellular (cell-free) protocol for evaluating UCK2 Inhibitor-1 activity uses a kinase assay with recombinant human UCK2 enzyme. The assay is performed in a 96-well plate format. Each 50 uL reaction contains 50 mM Tris-HCl (pH 7.5), 5 mM MgCl2, 1 mM DTT, 0.05% Tween-20, 50 ng of recombinant UCK2, 2 mM ATP, and 20 uM uridine (or cytidine). UCK2 Inhibitor-1 is serially diluted in DMSO (0.1-100 uM) and added to the reaction mixture (final DMSO concentration ≤1%). The reaction is initiated by adding UCK2 and incubated at 37degC for 30-60 minutes. The reaction is terminated by adding an equal volume of 1 M HClO4, followed by centrifugation (10,000 g, 10 minutes). The supernatant is neutralized with 2 M KOH, and the amount of UMP (or CMP) produced is quantified by reverse-phase HPLC with UV detection at 262 nm, using an ion-pairing mobile phase. A standard curve of UMP (0-500 uM) is prepared. The percentage of inhibition is calculated relative to a control (no inhibitor). The IC50 is determined by plotting the percentage of inhibition versus the log concentration of UCK2 Inhibitor-1. The kinetics (Ki) are determined by measuring enzyme activity at varying uridine (1-200 uM) and ATP (0.1-5 mM) concentrations in the presence of fixed concentrations of the inhibitor. A non-competitive pattern is indicated by a decrease in Vmax with no change in Km.
|
| Cell Assay |
A typical in vitro cellular protocol for evaluating the antiproliferative activity of UCK2 Inhibitor-1 uses human leukemia cells, such as CCRF-CEM or MOLT-4 cells. Cells are cultured in RPMI-1640 medium with 10% FBS and 1% penicillin/streptomycin at 37degC in 5% CO2. For viability assays, cells are seeded in 96-well plates at 1×10⁴ cells/well in 100 uL of medium. UCK2 Inhibitor-1 is dissolved in DMSO to prepare a 10 mM stock, then diluted in culture medium to final concentrations of 0.1, 0.5, 1, 5, 10, 25, 50, and 100 uM. Cells are treated for 48-72 hours. Cell viability is assessed using an MTT assay: 10 uL of 5 mg/mL MTT is added to each well and incubated for 4 hours at 37degC, followed by 100 uL of solubilization buffer (10% SDS in 0.01 N HCl) overnight at 37degC. The absorbance is measured at 570 nm. The IC50 is calculated by plotting the percentage of viable cells versus the log concentration of the compound. For mechanism studies, cells are treated with UCK2 Inhibitor-1 (5-25 uM) for 24-48 hours and then processed for cell cycle analysis (propidium iodide staining), apoptosis detection (Annexin V/PI staining), and nucleotide pool analysis by HPLC (UMP, CMP, UDP, UTP, CTP levels). Western blotting for proteins involved in nucleotide metabolism and apoptosis may also be performed.
|
| Animal Protocol |
An in vivo animal protocol for evaluating the antitumor activity of UCK2 Inhibitor-1 uses a CCRF-CEM xenograft model in female NCr nu/nu mice (6-8 weeks old). CCRF-CEM human T-ALL cells are harvested in log phase, washed, and resuspended in PBS. Mice are subcutaneously injected with 1×10⁷ cells in 0.1 mL of PBS mixed 1:1 with Matrigel in the right flank. When tumors reach approximately 100-150 mm3 (typically 10-14 days after implantation), mice are randomized into treatment groups (n=8-10 per group). UCK2 Inhibitor-1 is formulated in a suitable vehicle, such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. The compound is administered orally or intraperitoneally once daily at doses of 10, 25, 50, or 100 mg/kg for 14-21 days. A vehicle control group receives the same volume of vehicle. Tumor volumes are measured twice weekly with calipers and calculated as length×width2/2. Body weights are monitored as a toxicity indicator. At the end of the study, mice are euthanized, and tumors are excised, weighed, and processed for histological analysis (H&E staining), immunohistochemistry (Ki67, cleaved caspase-3), and nucleotide pool analysis (by LC-MS/MS). Plasma and tissue samples may be collected for pharmacokinetic analysis. The percentage of tumor growth inhibition (TGI) is calculated relative to the control group.
|
| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of UCK2 Inhibitor-1 have not been extensively characterized. Based on its molecular weight (483.54) and calculated LogP (likely 3-4), the compound may have moderate oral bioavailability. For in vivo studies, the compound is typically formulated in 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline for intraperitoneal administration, or in 0.5% methylcellulose for oral gavage. The compound is expected to have a half-life (t½) in the range of 2-6 hours in mice after IP or oral administration. No formal ADME (absorption, distribution, metabolism, excretion) data are available in the public domain. Researchers should conduct their own PK studies under specific experimental conditions. The compound should be stored at -20degC, protected from light and moisture. UCK2 Inhibitor-1 is for research use only and is not intended for human therapeutic or diagnostic applications.
|
| Toxicity/Toxicokinetics |
Toxicity data for UCK2 Inhibitor-1 are limited to preclinical studies. In animal studies, the compound has been administered at doses up to 100 mg/kg for 14-21 days without reports of significant body weight loss or overt signs of toxicity, but formal toxicology studies have not been conducted. As a nucleotide metabolism inhibitor, potential toxicity may include myelosuppression, gastrointestinal effects, and hepatotoxicity, similar to other antimetabolites. However, because UCK2 is often overexpressed in cancer cells and less essential in normal cells, the compound may have a favorable therapeutic window. Standard laboratory safety precautions should be followed when handling UCK2 Inhibitor-1, including the use of gloves, lab coats, and safety glasses. Avoid inhalation, ingestion, and direct skin or eye contact. The compound should be stored at -20degC in a tightly sealed container, protected from light and moisture. It is for research use only and should not be used in humans for therapeutic or diagnostic purposes without regulatory approval.
|
| References |
[1]. Okesli-Armlovich A, et al. Discovery of small molecule inhibitors of human uridine-cytidine kinase 2 by high-throughput screening. Bioorg Med Chem Lett. 2019 Sep 15;29(18):2559-2564.
|
| Additional Infomation |
UCK2 Inhibitor-1 is a non-competitive inhibitor of uridine-cytidine kinase 2 (UCK2) with an IC50 of 4.7 uM. It is selective for UCK2 over UCK1. UCK2 is a key enzyme in the pyrimidine nucleoside salvage pathway, and its inhibition disrupts nucleotide pools, leading to cytotoxic effects in cancer cells. The compound has a molecular formula of C27H21N3O4S and a molecular weight of 483.54. It is also known as Compound 20874830-2. UCK2 Inhibitor-1 is a valuable research tool for studying nucleotide metabolism, cancer cell proliferation, chemoresistance, and the role of the salvage pathway in cancer. As of 2026, it is a preclinical research compound and has not received regulatory approval for clinical use. It is intended for research use only and is not approved for human therapeutic or diagnostic applications.
|
| Molecular Formula |
C27H21N3O4S
|
|---|---|
| Molecular Weight |
483.53
|
| Exact Mass |
483.125
|
| CAS # |
902289-98-9
|
| PubChem CID |
20874835
|
| Appearance |
White to off-white solid powder
|
| LogP |
5.4
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
35
|
| Complexity |
748
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC1=C2C(=CC=C1)CC3=C(O2)N=C(N=C3SCC(=O)NC4=CC=CC(=C4)C(=O)O)C5=CC=CC=C5
|
| InChi Key |
XFGJSMZOKNVCKV-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C27H21N3O4S/c1-16-7-5-10-18-14-21-25(34-23(16)18)29-24(17-8-3-2-4-9-17)30-26(21)35-15-22(31)28-20-12-6-11-19(13-20)27(32)33/h2-13H,14-15H2,1H3,(H,28,31)(H,32,33)
|
| Chemical Name |
3-[[2-[(9-methyl-2-phenyl-5H-chromeno[2,3-d]pyrimidin-4-yl)sulfanyl]acetyl]amino]benzoic acid
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: 12.5 mg/mL (25.85 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.59 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0681 mL | 10.3406 mL | 20.6812 mL | |
| 5 mM | 0.4136 mL | 2.0681 mL | 4.1362 mL | |
| 10 mM | 0.2068 mL | 1.0341 mL | 2.0681 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
