Physicochemical Properties
| Molecular Formula | C30H34N8O3 |
| Molecular Weight | 554.64 |
| CAS # | 3048537-58-9 |
| Appearance | Typically exists as solids at room temperature |
| 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) |
Biological Activity
| Targets | HPK1 <0.51 nM (IC50) |
| ln Vitro | HPK1-IN-55 (0.5-10000 nM, 5 h) exhibits HPK1 enzyme inhibition in human PBMCs with an IC50 of <0.51 nM and inhibits cellular IL-2 secretion with an EC50 of 43.3 nM[1]. HPK1-IN-55 (0.001-100 nM) stimulates the release of IL-2 and IFN-γ in human PBMCs with EC50 values of 38.8 and 49.2 nM, respectively[1]. HPK1-IN-55 (0.00457-10 μM, 72 h) can promote T cell proliferation at low, medium and high concentrations in human immune T cells[1]. |
| ln Vivo | HPK1-IN-55 (1.5-12 mg/kg, po, bid, 5 weeks) has good antitumor effects as a monotherapy and shows additive effects in the CT26 model and synergistic effects in the MC38 model when used in combination with anti-PD-1[1]. HPK1-IN-55 (1 mg/kg, iv, 2 mg/kg, po) showed moderate clearance (Clp = 11.41 mL/min/kg), good oral exposure (DNAUC (0−24 h) =560.5 h•ng/mL) and bioavailability (F % = 42.0) in monkeys[1]. HPK1-IN-55 (1.5-12 mg/kg, po, bid, 5 weeks) showed good target engagement in the CT26 model[1]. In vivo pharmacokinetic parameters of HPK1-In-55 HPK1-IN-55 beagle dogs IV Clp (mL/min/kg) Vd (L/kg) 11.44 5.58 beagle dogs PO DNAUC (0-24 h) (h・ng/ml) t1/2 (h) F (%) 563 7.65 41.5 cyno monkeys IV Clp (mL/min/kg) Vd (L/kg) 11.41 5.07 cyno monkeys PO DNAUC (0-24 h) (h・ng/ml) t1/2 (h) F (%) 560.5 8.73 42 |
| Cell Assay |
Cell Proliferation Assay [1] Cell Types: human pan T cells Tested Tested Concentrations: 0.00457, 0.01372, 0.04115, 0.12346, 0.37037, 1.11111, 3.33333, 10 μM Incubation Duration: 72 h Experimental Results: Increased T cell proliferation at low, intermediate and high Tested Tested Concentrationss in human pan T cells. |
| Animal Protocol |
Animal/Disease Models: Balb/c mice bearing the CT26 murine colorectal cancer tumor[1] . Doses: 3, 10, or 30 mg/kg Route of Administration: p.o., b.i.d, 24 h Experimental Results: Animals receiving a 10 and 30 mg/kg dose exhibited a free plasma concentration that inhibited pSLP76 > 50% in the spleen for more than 6 h, whereas animals receiving the 3 mg/kg dose did not exhibit this sustained reduction. Animal/Disease Models: Balb/c mice injected with CT26 cells[1] . Doses: 1.5, 12 mg/kg Route of Administration: p.o., b.i.d, 5weeks Experimental Results: Exhibited a good antitumor response with a tumor growth inhibition (TGI) value of 64.3%. Animal/Disease Models: Balb/c mice injected with MC38 cells[1] . Doses: 3, 12 mg/kg Route of Administration: p.o., b.i.d, 5weeks Experimental Results: Demonstrated similar antitumor efficacy in the MC38 model with a TGI value of 34.9% for the 3 mg/kg group and 59.4% for the 12 mg/kg group. |
| References |
[1]. Peng J, et.al. Discovery of Pyridine-2-Carboxamides Derivatives as Potent and Selective HPK1 Inhibitors for the Treatment of Cancer. J Med Chem. 2024 Dec 12;67(23):21520-21544. |
Solubility Data
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8030 mL | 9.0149 mL | 18.0297 mL | |
| 5 mM | 0.3606 mL | 1.8030 mL | 3.6059 mL | |
| 10 mM | 0.1803 mL | 0.9015 mL | 1.8030 mL |