| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
IC50: 142 nM (Btk)[1]
BTK inhibitor 18 targets Bruton's tyrosine kinase (Btk), a Tec family non-receptor kinase that plays a critical role in B cell receptor (BCR) signaling. It acts as an irreversible covalent inhibitor, binding to a noncatalytic cysteine residue (Cys481) in the active site of Btk [2L12-L14]. By forming a permanent bond, it provides sustained inhibition of Btk-mediated signaling pathways, which include NF-kappaB, MAP kinase, and PI3K/AKT pathways. |
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| ln Vitro |
Compound 27, also known as BTK inhibitor 18, targets a noncatalytic cysteine residue (Cys481) for the creation of covalent bonds, therefore irreversibly inhibiting BTK[1]. With an IC50 of 84 nM, BTK inhibitor 18 (Compound 27) prevents anti-IgM-induced B cell activation in human whole blood[1]. Additionally, BMX, LCK, ErbB4, TEC, and TXK kinases are inhibited by BTK inhibitor 18 (Compound 27), with IC50 values of 129 nM, 130 nM, 377 nM, 409 nM, and 1770 nM, respectively[1].
In vitro, BTK inhibitor 18 is a potent Btk inhibitor with an IC₅0 of 142 nM [12L7]. It has been shown to inhibit anti-IgM-induced activation of B cells, effectively blocking downstream signaling events such as calcium flux and phosphorylation of PLCgamma2 [2L33-L34]. It also possesses anti-inflammatory activities, indicating its potential to suppress the function of inflammatory cells like macrophages and mast cells. |
| ln Vivo |
In a rat collagen-induced arthritis model, BTK inhibitor 18 (Compound 27; 1-30 mg/kg; oral administration; once daily; for 7 days) therapy demonstrates dose-dependent efficacy at reducing joint inflammation[1]. BTK inhibitor 18 (Compound 27) pharmacokinetics in IV and PO are studied in fasted dogs (0.5 and 2.5 mg/kg IV and PO) and nonfasted rats (1 and 5 mg/kg IV and PO). Rat and dog IV pharmacokinetics are characterized by low clearance in the former and moderate clearance in the latter, as well as a moderate volume of distribution and short plasma half-lives (T1/2 of 1.9 h for dog and 0.3 h for rat, respectively). In rats and dogs, the oral bioavailability is 30% and 68%, respectively[1].
In vivo, BTK inhibitor 18 has demonstrated anti-inflammatory effects. A study using a rat collagen-induced arthritis (CIA) model evaluated the compound. Animals were administered the inhibitor orally at doses of 1 mg/kg, 3 mg/kg, 10 mg/kg, and 30 mg/kg once daily for 7 days [2L4-L6]. The results indicated a dose-dependent improvement in arthritis scores, confirming the compound's ability to target Btk-mediated inflammation in a whole-body system. |
| Enzyme Assay |
BTK inhibitor 18's irreversible binding is typically assessed using biochemical kinase assays. Purified recombinant human Btk enzyme is incubated with various concentrations of the inhibitor in the presence of ATP and a specific peptide substrate. The reaction is initiated, and the remaining kinase activity is measured. The IC₅0 is calculated from the inhibition curve. Irreversible binding can be confirmed via a dilution or dialysis experiment, showing that the effect is long-lasting after the inhibitor is removed.
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| Cell Assay |
For cell-based assays, a human B cell lymphoma cell line, such as Ramos or Raji cells, is used. Cells are pre-treated with varying concentrations of BTK inhibitor 18 for 1-2 hours, then stimulated via the BCR (e.g., with anti-IgM antibody) to activate Btk. Endpoints include measuring phosphorylation of Btk (Y223) and its downstream substrate PLCgamma2 (Y759) by Western blot. Additionally, the inhibition of B cell activation can be measured by flow cytometry to assess the downregulation of activation markers like CD86.
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| Animal Protocol |
Animal/Disease Models: A rat collagen-induced arthritis (CIA) model[1]
Doses: 1 mg/kg, 3 mg/kg, 10 mg/kg, 30 mg/kg Route of Administration: Oral administration ; one time/day; for 7 days Experimental Results: Attenuated hind paw inflammation in a dose-dependent manner. In vivo efficacy is commonly assessed using a rat collagen-induced arthritis (CIA) model. Female Lewis rats are immunized with type II collagen to induce arthritis. Once arthritis is established, BTK inhibitor 18 is administered orally at doses such as 1, 3, 10, and 30 mg/kg once daily for 7 days [2L4-L6]. Arthritis severity is scored visually every 2-3 days. At the end of the study, paw swelling is measured, and joint tissue is collected for histopathological analysis of inflammation and bone erosion. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of BTK inhibitor 18 have been studied. It is characterized as an orally active compound, indicating good oral bioavailability [12L6]. In pharmacokinetic studies, the compound was administered to fasted dogs via IV (0.5 mg/kg) and PO (2.5 mg/kg) routes to determine its absorption, clearance, and half-life [2L15-L16]. These studies confirmed that it is well-absorbed after oral administration and has a PK profile suitable for a once-daily oral drug.
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| Toxicity/Toxicokinetics |
Comprehensive toxicological data for BTK inhibitor 18 are not detailed in standard product literature. As a research-use compound that forms an irreversible covalent bond, it has potential for off-target toxicity if it reacts with other cysteine-containing proteins. Standard safety assessments for acute toxicity, genotoxicity, and organ-specific toxicity would be required for clinical development. For research use, standard chemical safety precautions for kinase inhibitors should be followed.
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| References | |
| Additional Infomation |
BTK inhibitor 18 (Compound 27, also known as CHMFL-BTK-11) has the molecular formula C2₉H2₅N₅O4S2 and a molecular weight of 571.67 g/mol [13L6]. The CAS number is not specified in the datasheet. It has a purity of ≥98%. The powder can be stored at -20degC for up to 3 years or at 4degC for 2 years; in solution, it can be stored at -80degC for 6 months or at -20degC for 1 month [13L8-L9].
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| Molecular Formula |
C29H25N5O4S2
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| Molecular Weight |
571.67
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| Appearance |
Typically exists as solid at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 1.7493 mL | 8.7463 mL | 17.4926 mL | |
| 5 mM | 0.3499 mL | 1.7493 mL | 3.4985 mL | |
| 10 mM | 0.1749 mL | 0.8746 mL | 1.7493 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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