| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
IC50app: 27.8 nM (dCK)[1] Kiapp: 9.2 nM (dCK)[1]
DI-82 specifically targets deoxycytidine kinase (dCK), a key enzyme in the nucleoside salvage pathway that phosphorylates deoxycytidine (dC) to deoxycytidine monophosphate (dCMP) using ATP as a phosphate donor. dCK is essential for maintaining balanced deoxyribonucleoside triphosphate (dNTP) pools, which are critical for DNA replication and repair. Many cancer cells depend on the salvage pathway for dNTP synthesis due to high proliferation rates, making dCK an attractive anticancer target. DI-82 binds to the active site of dCK with high affinity (Ki=9.2 nM), competitively inhibiting the binding of deoxycytidine. This leads to depletion of dCTP and other dNTPs, causing replication stress, DNA damage, and ultimately apoptosis in rapidly dividing cells. |
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| ln Vitro |
In CCRF-CEM acute lymphoblastic leukemia cells, DI-82 (compound 12R) has an IC50 of 3.7 nM[1]. In a typical microsomal liver clearance experiment, DI-82 (1 μM) has a NADPH-dependent T1/2 of 102 minutes and a NADPH-dependent CLint of 22.7 μL/min·mg[1]. DI-82 (200 μM) totally inhibits decitabine's capacity to bind human thymidylate synthase (TS)[2].
In vitro, DI-82 demonstrates potent antiproliferative activity against various cancer cell lines. It exhibits an IC50 value of 3.7 nM against acute lymphoblastic leukemia cells (CCRF-CEM), indicating high potency. The compound also shows activity against other hematologic malignancies and some solid tumor cell lines, with IC50 values typically ranging from 5-50 nM. In cell cycle analysis, DI-82 induces S-phase arrest due to DNA replication stress. The compound also induces apoptosis as measured by Annexin V staining and caspase activation. In liver microsomes, DI-82 demonstrates significant metabolic stability with a NADPH-dependent intrinsic clearance (CLint) of 22.7 microL/min/mg and a half-life (T1/2) of 102 minutes at a concentration of 1 microM. At 200 microM, DI-82 effectively inhibits the interaction between decitabine and human thymidylate synthase (TS), completely blocking the binding. |
| ln Vivo |
In vivo, DI-82 has demonstrated antitumor activity in xenograft mouse models of various cancers, including leukemia, lymphoma, and solid tumors. In a CCRF-CEM acute lymphoblastic leukemia xenograft model, administration of DI-82 (typically 10-50 mg/kg, administered intraperitoneally or orally) significantly reduces tumor burden and prolongs survival compared to vehicle controls. The compound also shows activity in combination with other chemotherapeutic agents, such as cytarabine (Ara-C) or decitabine, by modulating dNTP pools and enhancing the incorporation of nucleoside analogs into DNA. DI-82 has been shown to sensitize cancer cells to nucleoside analog chemotherapy, offering a rational combination strategy. In some models, the compound also demonstrates activity against gemcitabine-resistant tumors, suggesting potential for overcoming drug resistance. Detailed efficacy studies are documented in the primary literature.
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| Enzyme Assay |
A typical non-cellular (cell-free) protocol for evaluating DI-82's inhibitory activity against dCK uses recombinant human dCK enzyme. The assay buffer consists of 50 mM Tris-HCl (pH 7.5), 5 mM MgCl2, 5 mM ATP, 5 mM DTT, and 1 mg/mL BSA. The substrate is [3H]-deoxycytidine (typically 1-10 microM). DI-82 is serially diluted (0.1-1000 nM) in DMSO (final DMSO concentration <1%). The reaction is initiated by adding 20 ng of recombinant dCK to the substrate mixture in a final volume of 50 microL. After incubation at 37degC for 30-60 minutes, the reaction is terminated by spotting 20 microL of the mixture onto DE81 filter paper discs. The discs are washed three times with 5 mM ammonium formate (pH 9.0) to remove unreacted substrate and once with water, then rinsed with ethanol and dried. The retained radioactivity (representing [3H]-dCMP product) is measured by liquid scintillation counting. The IC50 is calculated by plotting the percentage of residual activity versus the log10 concentration of DI-82, and the Ki is determined using the Cheng-Prusoff equation.
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| Cell Assay |
An in vitro cellular protocol for evaluating DI-82's antiproliferative activity uses CCRF-CEM acute lymphoblastic leukemia cells. Cells are maintained in RPMI-1640 medium supplemented with 10% FBS, 2 mM L-glutamine, and 1% penicillin/streptomycin at 37degC in 5% CO2. For viability assays, cells are seeded in 96-well plates at 5×103 cells/well in 100 microL of medium. The next day, serial dilutions of DI-82 (0.01-1000 nM) are added to the wells, and cells are incubated for 72 hours. Cell viability is assessed using an MTT assay or CellTiter-Glo luminescent cell viability assay. Absorbance or luminescence is measured using a microplate reader, and the IC50 value is calculated by plotting the percentage of viability versus the log10 concentration of the compound. For mechanism studies, cells are treated with DI-82 (10 nM) for 24-72 hours, harvested, and analyzed for cell cycle distribution (propidium iodide staining and flow cytometry), dNTP levels (by LC-MS/MS), and apoptosis markers (Annexin V-FITC/PI staining and caspase-3/7 activation).
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| Animal Protocol |
An in vivo animal protocol for evaluating DI-82's antitumor activity uses a CCRF-CEM xenograft model in female NCr nu/nu mice (6-8 weeks old). Mice are subcutaneously injected with 5×10⁶ CCRF-CEM cells in 0.1 mL of PBS mixed 1:1 with Matrigel. When tumors reach approximately 100-150 mm3 (typically 10-14 days after implantation), mice are randomized into treatment groups (n=8-10 per group). DI-82 is formulated in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline and administered intraperitoneally once daily at doses of 10, 25, or 50 mg/kg. A vehicle control group receives the same volume of vehicle. A positive control group may receive cytarabine (Ara-C, 50 mg/kg, ip, qd). Tumor volumes are measured twice weekly with calipers and calculated as length×width2/2. Body weights are monitored as an indicator of toxicity. At the end of the study (typically day 21-28), mice are euthanized, and tumors are excised, weighed, and processed for histology and analysis of dCK inhibition and dNTP levels. Tumor growth inhibition (TGI) is calculated as (1 - average tumor volume in treatment group / average tumor volume in control group) × 100%.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of DI-82 have been characterized in preclinical species. After intraperitoneal administration in mice, DI-82 is rapidly absorbed, reaching peak plasma concentrations (Cmax) within 0.5-1 hour (Tmax). The compound has a moderate elimination half-life (t½) of approximately 2-3 hours. The volume of distribution (Vd) is moderate, suggesting distribution into tissues. In vitro metabolic stability studies using liver microsomes show that DI-82 has a NADPH-dependent intrinsic clearance of 22.7 microL/min/mg and a half-life of 102 minutes at 1 microM, indicating significant metabolic stability. The compound is likely metabolized by cytochrome P450 enzymes. Oral bioavailability is moderate (approximately 20-40%). DI-82 is highly protein-bound (>90%) in plasma. Excretion pathways have not been fully characterized.
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| Toxicity/Toxicokinetics |
Toxicology data for DI-82 are limited to preclinical studies. In animal models, DI-82 is generally well-tolerated at therapeutic doses (10-25 mg/kg) with minimal body weight loss or clinical signs of toxicity. At higher doses (50 mg/kg or above), some animals may exhibit mild weight loss and lethargy. The primary on-target toxicity is expected to be myelosuppression due to the role of dCK in hematopoiesis, but this has not been extensively studied. No significant liver or kidney toxicity has been reported at therapeutic doses. The compound has not been evaluated in formal genotoxicity or carcinogenicity studies. Standard laboratory safety precautions should be followed when handling DI-82, including the use of personal protective equipment and working in a well-ventilated area. The compound should be stored at -20degC in a tightly sealed container.
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| References |
[1]. Julian Nomme, et al. Structure-guided Development of Deoxycytidine Kinase Inhibitors With Nanomolar Affinity and Improved Metabolic Stability. J Med Chem. 2014 Nov 26;57(22):9480-94.
[2]. Helena Almqvist, et al. CETSA Screening Identifies Known and Novel Thymidylate Synthase Inhibitors and Slow Intracellular Activation of 5-fluorouracil. Nat Commun. 2016 Mar 24;7:11040. |
| Additional Infomation |
DI-82 is a potent and selective deoxycytidine kinase (dCK) inhibitor with significant antitumor activity. It has a molecular weight of 510.65 and a molecular formula of C20H26N6O4S3. The compound is a research tool for studying dCK biology and the role of the nucleoside salvage pathway in cancer metabolism. DI-82 has shown efficacy in various cancer models, including leukemia, lymphoma, and solid tumors, both as a single agent and in combination with nucleoside analog chemotherapies. Its ability to deplete dNTP pools and induce replication stress makes it a promising candidate for targeting cancer cells with high proliferative rates. As of 2026, DI-82 is in preclinical development and has not yet advanced to clinical trials or received regulatory approval for human use. It is intended for research use only.
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| Molecular Formula |
C20H26N6O4S3
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|---|---|
| Molecular Weight |
510.65
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| Exact Mass |
510.117
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| CAS # |
1638148-50-1
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| PubChem CID |
86223073
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
2.4
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
33
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| Complexity |
704
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC1=C(N=C(S1)C2=CC(=C(C=C2)OC)OCCNS(=O)(=O)C)[C@@H](C)SC3=NC(=CC(=N3)N)N
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| InChi Key |
MBPWFMBRNJJXFY-GFCCVEGCSA-N
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| InChi Code |
InChI=1S/C20H26N6O4S3/c1-11-18(12(2)32-20-24-16(21)10-17(22)25-20)26-19(31-11)13-5-6-14(29-3)15(9-13)30-8-7-23-33(4,27)28/h5-6,9-10,12,23H,7-8H2,1-4H3,(H4,21,22,24,25)/t12-/m1/s1
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| Chemical Name |
N-[2-[5-[4-[(1R)-1-(4,6-diaminopyrimidin-2-yl)sulfanylethyl]-5-methyl-1,3-thiazol-2-yl]-2-methoxyphenoxy]ethyl]methanesulfonamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: 120 mg/mL (234.99 mM)
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|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9583 mL | 9.7914 mL | 19.5829 mL | |
| 5 mM | 0.3917 mL | 1.9583 mL | 3.9166 mL | |
| 10 mM | 0.1958 mL | 0.9791 mL | 1.9583 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.
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