| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
hematopoietic progenitor kinase 1 (HPK1)[1]
HPK1-IN-19 specifically targets Hematopoietic Progenitor Kinase 1 (HPK1). HPK1 is a negative regulator of T-cell receptor (TCR) signaling. By blocking HPK1 activity, the compound has the potential to enhance T-cell activation, cytokine production, and anti-tumor immune responses. This mechanism is promising for cancer immunotherapy research by boosting immune cell function and overcoming tumor-induced immunosuppression. |
|---|---|
| ln Vitro |
Specific in vitro activity data for HPK1-IN-19, such as an IC50 value, is not provided in standard chemical databases. However, it is described as a potent and selective inhibitor of HPK1, a negative regulator of T-cell receptor (TCR) signaling. Its selectivity is based on its chemical structure as described in the patent literature. It is expected to enhance T-cell activation and cytokine production in cell-based assays.
|
| ln Vivo |
Specific in vivo activity data for HPK1-IN-19 has not been published. Based on its described mechanism as an HPK1 inhibitor, it is hypothesized to enhance the anti-tumor immune response in vivo. This could be demonstrated in murine syngeneic tumor models (e.g., CT26 colon carcinoma, B16-F10 melanoma). HPK1-IN-19 would be predicted to increase CD8+ T-cell infiltration into tumors and augment the efficacy of checkpoint inhibitors such as anti-PD-1.
|
| Enzyme Assay |
The specific protocol for evaluating HPK1-IN-19 likely involves a standard radiometric kinase activity assay. Recombinant human HPK1 kinase domain is incubated with a serial dilution of HPK1-IN-19 (1 nM to 100 uM) in the presence of 33P-gamma-ATP and a protein substrate such as myelin basic protein (MBP). Following incubation at 30degC for 30 minutes, the reaction is stopped and spotted onto P81 phosphocellulose paper. Unincorporated radioactivity is removed by multiple washes in phosphoric acid. The incorporated radioactivity is measured by liquid scintillation counting. IC50 values are determined from concentration-response curves.
|
| Cell Assay |
In a typical in vitro cell-based assay, primary human CD4+ T cells are isolated from healthy donor PBMCs using magnetic negative selection. Cells are activated with plate-bound anti-CD3 (1 ug/mL) and soluble anti-CD28 (1 ug/mL) antibodies. HPK1-IN-19 is added at concentrations ranging from 1 nM to 10 uM. After 72 hours, cell culture supernatants are harvested, and IL-2 and IFN-gamma levels are measured using a sandwich ELISA. T-cell proliferation is assessed by CFSE dilution assay using flow cytometry, analyzing dye dilution in the CD8+ and CD4+ T-cell populations.
|
| Animal Protocol |
An in vivo protocol for HPK1-IN-19 would involve the MC38 syngeneic colon carcinoma model in female C57BL/6 mice. Mice are inoculated subcutaneously with 5e5 MC38 cells. Once tumors reach a mean size of approximately 100 mm3, mice are randomized into treatment groups. HPK1-IN-19 is administered orally at doses of 10-50 mg/kg once or twice daily. Control groups receive vehicle or an isotype antibody. Tumor volumes are measured every 2-3 days. At the end of the study, spleens and tumors are collected for immunophenotyping by flow cytometry to evaluate tumor-infiltrating lymphocyte (TIL) populations, especially CD8+ T-cell activation markers.
|
| ADME/Pharmacokinetics |
Detailed pharmacokinetic parameters for HPK1-IN-19 are not available. As a small molecule HPK1 inhibitor extracted from patent literature, a standard study would involve cassette dosing in male BALB/c mice. The compound would be administered via oral gavage (e.g., 10 mg/kg) and intravenous injection (1 mg/kg). Serial plasma samples would be collected at various time points. HPK1-IN-19 concentrations would be determined by a qualified LC-MS/MS method. Pharmacokinetic parameters, including T1/2, Cmax, AUC, clearance, and oral bioavailability (%F), would be calculated using non-compartmental analysis.
|
| Toxicity/Toxicokinetics |
Toxicology data for HPK1-IN-19 is not available. As an inhibitor of HPK1, which acts as a negative regulator of the immune system, the primary on-target toxicity is expected to be immune-related. This could include autoimmune-like pathology or cytokine release syndrome at excessively high exposures. Standard safety assessment would include in vitro hERG channel inhibition testing, CYP450 inhibition screening, and an exploratory 7-day repeat-dose oral toxicity study in rats to identify any target organ toxicity.
|
| References | |
| Additional Infomation |
HPK1-IN-19 is a research-grade chemical and is not approved for clinical use. It is described in patent WO2018102366A1 as compound I-47. Its molecular formula is C27H32N7O2P, and it has a molecular weight of 517.56 with a purity of >98%. The compound is stored as a powder at -20degC. It is a valuable tool for studying HPK1's role as a negative regulator of the immune system in cancer immunotherapy research.
|
| Molecular Formula |
C27H32N7O2P
|
|---|---|
| Molecular Weight |
517.562445640564
|
| Exact Mass |
517.235
|
| CAS # |
2227609-33-6
|
| PubChem CID |
134564648
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
3.3
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
37
|
| Complexity |
799
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
N(C1C=CC=CC=1P(=O)(C)C)C1=NC(NC2C=C3CN(C)CCC3=CC=2OC)=NC=C1C1C=NN(C)C=1
|
| InChi Key |
AZENVIBXSMLBQC-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C27H32N7O2P/c1-33-11-10-18-13-24(36-3)23(12-19(18)16-33)31-27-28-15-21(20-14-29-34(2)17-20)26(32-27)30-22-8-6-7-9-25(22)37(4,5)35/h6-9,12-15,17H,10-11,16H2,1-5H3,(H2,28,30,31,32)
|
| Chemical Name |
4-N-(2-dimethylphosphorylphenyl)-2-N-(6-methoxy-2-methyl-3,4-dihydro-1H-isoquinolin-7-yl)-5-(1-methylpyrazol-4-yl)pyrimidine-2,4-diamine
|
| 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 |
| 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) |
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
|
|---|---|
| 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 | 1.9321 mL | 9.6607 mL | 19.3214 mL | |
| 5 mM | 0.3864 mL | 1.9321 mL | 3.8643 mL | |
| 10 mM | 0.1932 mL | 0.9661 mL | 1.9321 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
