| Size | Price | Stock | Qty |
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| Targets |
The primary targets of KDM4-IN-2 are the KDM4 and KDM5 subfamilies of Jumonji C (JmjC) domain-containing histone lysine demethylases. Specifically, it inhibits KDM4A with a Ki of 4 nM and KDM5B with a Ki of 7 nM. These enzymes are involved in the demethylation of histone H3 lysine 9 (H3K9) and histone H3 lysine 4 (H3K4), respectively. By inhibiting these enzymes, KDM4-IN-2 alters the methylation status of histones, leading to changes in chromatin structure and gene expression. The compound also shows activity against other KDM4 and KDM5 family members. Its high potency and selectivity make it a valuable tool for studying the role of these enzymes in various biological processes.
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| ln Vitro |
In vitro, KDM4-IN-2 is a potent and selective inhibitor of KDM4 and KDM5 histone demethylases. It exhibits Ki values of 4 nM for KDM4A and 7 nM for KDM5B in biochemical assays. The compound's inhibitory activity is concentration-dependent, with effects observed at nanomolar concentrations. It shows selectivity for KDM4 and KDM5 over other histone demethylases and epigenetic enzymes. Its activity can be assessed using cell-free enzyme assays, where it inhibits the demethylation of histone substrates. KDM4-IN-2 is a valuable tool for studying the role of histone demethylases in gene regulation and for validating these enzymes as therapeutic targets.
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| ln Vivo |
In vivo, KDM4-IN-2 has potential applications in studying the role of histone demethylases in disease models. As a potent and selective inhibitor of KDM4 and KDM5, the compound can be used to modulate gene expression in animal models of cancer, neurological disorders, and other diseases. Its in vivo efficacy would depend on its pharmacokinetic properties, including bioavailability, half-life, and tissue distribution. Further studies are needed to evaluate its in vivo activity, efficacy, and safety. The compound is a valuable tool for preclinical studies of histone demethylase biology.
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| Enzyme Assay |
The in vitro enzyme inhibition activity of KDM4-IN-2 can be assessed using cell-free assays with recombinant KDM4A or KDM5B enzymes. A typical protocol involves incubating the enzyme with a histone peptide substrate (e.g., H3K9me3 or H3K4me3), alpha-ketoglutarate, Fe2+, and KDM4-IN-2 at various concentrations in a reaction buffer. The reaction is carried out at 37degC for a specified period, and the amount of demethylated product is measured using a coupled assay or by detecting the release of formaldehyde or succinate. The Ki value is determined by fitting the data to appropriate kinetic models. The selectivity of the compound can be assessed by testing its activity against a panel of related histone demethylases.
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| Cell Assay |
For in vitro cellular experiments, cancer cell lines or other cell types are cultured in appropriate media and treated with KDM4-IN-2 at various concentrations (typically 0.1-100 microM). After treatment, cells are harvested, and histone methylation levels are assessed by Western blot using antibodies specific for methylated histone marks (e.g., H3K9me3, H3K4me3). Gene expression changes are assessed by qRT-PCR or RNA-seq. Cell proliferation is measured using MTT or other cell proliferation assays. The duration of treatment varies depending on the experimental design but typically ranges from 24 to 72 hours. The compound's effects on cell cycle and apoptosis can also be assessed.
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| Animal Protocol |
In vivo animal experiments with KDM4-IN-2 would typically involve oral or intraperitoneal administration in mice or rats. A common dosing regimen would be based on pharmacokinetic studies to determine the optimal dose and route of administration. For efficacy studies, the compound is administered to animal models of cancer or other diseases, and disease progression is monitored. Histone methylation levels and gene expression changes are assessed in tissues. Pharmacokinetic and toxicity studies are also performed to characterize its safety profile. Further preclinical studies are needed to fully evaluate its in vivo activity.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for KDM4-IN-2 are not well-documented in the available literature. As a small molecule inhibitor with a molecular weight of 426.51 g/mol, it is expected to have moderate lipophilicity and may be orally bioavailable. The compound is typically stored lyophilized at -20degC and is stable for 36 months in lyophilized form. In solution, it should be stored at -20degC and used within 1-3 months to prevent loss of potency. Further studies are needed to determine its pharmacokinetic profile, including oral bioavailability, half-life, protein binding, and tissue distribution.
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| Toxicity/Toxicokinetics |
The toxicity profile of KDM4-IN-2 has not been extensively characterized. As a potent inhibitor of histone demethylases, which are involved in regulating gene expression, the compound may have potential toxicity due to off-target effects on gene expression. In vitro cytotoxicity studies would be needed to assess its effects on mammalian cell lines. In vivo toxicity studies in animal models would also be required to determine its safety profile. The compound should be handled with standard laboratory precautions and is intended for research use only.
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| References | |
| Additional Infomation |
KDM4-IN-2 (Compound 19a) is a potent and selective dual inhibitor of KDM4 and KDM5 histone lysine demethylases. It exhibits Ki values of 4 nM for KDM4A and 7 nM for KDM5B. The compound has a molecular weight of 426.51 g/mol and a molecular formula of C25H26N6O. KDM4-IN-2 is a valuable tool for studying the role of histone demethylases in gene regulation, cancer, and other diseases. It is available as a research compound and is not approved for clinical use.
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| Molecular Formula |
C25H26N6O
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| Molecular Weight |
426.513544559479
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| Exact Mass |
426.216
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| CAS # |
2369607-62-3
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| PubChem CID |
138393336
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.1
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
32
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| Complexity |
719
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1NC(=O)C2C=CN=C(N3C=C(CCN4CCC5(CC4)C4=C(C=CC=C4)CC5)C=N3)C=2N=1
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| InChi Key |
AVFIHINZEZRGBL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C25H26N6O/c32-24-20-6-11-26-23(22(20)27-17-28-24)31-16-18(15-29-31)7-12-30-13-9-25(10-14-30)8-5-19-3-1-2-4-21(19)25/h1-4,6,11,15-17H,5,7-10,12-14H2,(H,27,28,32)
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| Chemical Name |
8-[4-(2-spiro[1,2-dihydroindene-3,4'-piperidine]-1'-ylethyl)pyrazol-1-yl]-3H-pyrido[3,4-d]pyrimidin-4-one
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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 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)
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| Solubility (In Vitro) |
DMSO : ~4 mg/mL (~9.38 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 | 2.3446 mL | 11.7231 mL | 23.4461 mL | |
| 5 mM | 0.4689 mL | 2.3446 mL | 4.6892 mL | |
| 10 mM | 0.2345 mL | 1.1723 mL | 2.3446 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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