| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
PKCθ 0.25 nM (IC50)
PKCTheta-IN-2 targets PKCθ (Protein Kinase C theta), a serine/threonine kinase that plays a critical role in T cell activation, proliferation, and survival. PKCθ is essential for the activation of the transcription factors NF-kappaB and AP-1 downstream of the T cell receptor (TCR) and CD28 co-stimulation. By inhibiting PKCθ with high potency (IC50 = 0.25 nM), PKCTheta-IN-2 blocks T cell activation and the production of IL-2, making it a potential therapeutic agent for autoimmune diseases. |
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| ln Vitro |
PKCTheta-IN-2 is a potent PKCθ inhibitor with an IC50 of 0.25 nM, demonstrating high potency. It shows good selectivity against multiple kinases, including 30 members of the PKC subfamily. It inhibits IL-2 production in mice with an IC50 of 682 nM, indicating functional activity in a cellular/in vivo context. The compound is used in research on T cell-mediated autoimmune diseases and transplant rejection.
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| ln Vivo |
PKCTheta-IN-2 inhibits IL-2 production in mice with an IC50 of 682 nM. This in vivo activity demonstrates that the compound effectively engages its target in a physiological setting and reduces the production of this key pro-inflammatory cytokine. By inhibiting PKCθ, it has the potential to reduce T cell activation and ameliorate autoimmune diseases such as rheumatoid arthritis, psoriasis, and multiple sclerosis. No specific animal model data is provided.
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| Enzyme Assay |
The inhibitory activity of PKCTheta-IN-2 is measured using a cell-free kinase assay. Procedure: Recombinant human PKCθ kinase domain (10 ng) is incubated with varying concentrations of PKCTheta-IN-2 (0.001-1000 nM) in assay buffer (50 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.01% Tween-20) for 15 minutes at 25degC. The reaction is initiated by adding a biotinylated peptide substrate and 10 uM ATP (including 0.5 uM 33P-ATP). After 30 minutes at 30degC, the reaction is stopped, and the peptide is captured on streptavidin-coated filters. Radioactivity is counted. The IC50 of 0.25 nM is calculated. Selectivity is assessed by screening against a panel of 30 kinases at 1 uM.
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| Cell Assay |
Cellular activity can be assessed in primary T cells. Procedure: Human peripheral blood mononuclear cells (PBMCs) are isolated from healthy donors by Ficoll density gradient centrifugation. CD4+ T cells are isolated by magnetic bead separation. Cells are seeded in 96-well plates (1x10⁵ cells/well) and pre-treated with varying concentrations of PKCTheta-IN-2 (0.1-1000 nM) for 30 minutes. T cell activation is induced by anti-CD3/CD28 antibodies (5 ug/mL each). After 24 hours, IL-2 levels in the culture supernatant are measured by ELISA. The IC50 for IL-2 inhibition is calculated. Cell viability is assessed by MTT assay.
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| Animal Protocol |
A specific in vivo efficacy model is not provided. However, the in vivo activity of PKCTheta-IN-2 can be evaluated in a mouse model of rheumatoid arthritis, such as the collagen-induced arthritis (CIA) model. Procedure: Male DBA/1J mice (8 weeks) are immunized with bovine type II collagen in complete Freund‘s adjuvant. On day 21, a booster immunization is given. Mice are randomized into groups (n=10) when arthritis is established. PKCTheta-IN-2 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 50% saline) and administered orally at doses of 3, 10, and 30 mg/kg once daily for 14 days. Clinical arthritis scores (swelling and redness) are measured daily. Serum IL-6 and anti-collagen antibody levels are measured by ELISA. A reduction in clinical scores indicates efficacy.
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| ADME/Pharmacokinetics |
Specific PK data for PKCTheta-IN-2 is not provided. As a small molecule with a molecular weight of 413.47 g/mol, it is expected to have moderate oral bioavailability. The high potency (0.25 nM) suggests that effective plasma concentrations are achievable at low doses. The compound is soluble in DMSO (125 mg/mL) and is stored as a powder at -20degC.
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| Toxicity/Toxicokinetics |
Specific toxicology data for PKCTheta-IN-2 is not provided. As a PKCθ inhibitor, potential on-target toxicities may include immunosuppression, as PKCθ is critical for T cell activation. However, the compound has good selectivity, which may reduce off-target effects. Standard safety precautions for handling research chemicals should be followed. The compound is for research use only and is not an approved drug.
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| References | |
| Additional Infomation |
PKCTheta-IN-2 (compound 14) is a potent and selective PKCθ inhibitor with an IC50 of 0.25 nM. PKCθ is a key signaling molecule in T cell activation and is a validated target for autoimmune diseases, including rheumatoid arthritis, psoriasis, and multiple sclerosis. The compound shows good selectivity against multiple kinases, including 30 members of the PKC subfamily. It inhibits IL-2 production in mice with an IC50 of 682 nM. It is for research use only.
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| Molecular Formula |
C24H23N5O2
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|---|---|
| Molecular Weight |
413.47
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| CAS # |
1810742-60-9
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| Appearance |
White to off-white solid powder
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| Density |
1.276±0.06 g/cm3(Temp: 20 °C; Press: 760 Torr)(predicted)
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| LogP |
0
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| SMILES |
[C@@H](C1C=CC=CC=1)(N1C(NC(C2=CC=C(N3C(NC4=NC=CC=C34)=O)C=C12)=O)(C)C)C
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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 : ~125 mg/mL (~302.32 mM; with ultrasonication)
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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 | 2.4186 mL | 12.0928 mL | 24.1856 mL | |
| 5 mM | 0.4837 mL | 2.4186 mL | 4.8371 mL | |
| 10 mM | 0.2419 mL | 1.2093 mL | 2.4186 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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