| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
PRT-060318 dihydrochloride specifically targets and inhibits spleen tyrosine kinase (Syk), a key signaling molecule in the B cell receptor (BCR) pathway. By inhibiting Syk, it blocks downstream signaling cascades, including the PI3K/AKT and MAPK pathways, which are crucial for B cell survival, proliferation, and migration. It also inhibits platelet activation via GPVI/FcRgamma signaling, preventing immune complex-mediated platelet aggregation.
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| ln Vitro |
PRT-060318 (PRT318) (0-3 µM, 15 min) dihydrochloride completely inhibited platelet aggregation induced by heparin-induced thrombocytopenia (HIT) immune complexes in human and transgenic HIT mice [1].
In vitro, PRT-060318 dihydrochloride inhibits Syk activity with an IC50 of 3 nM. It suppresses the activation and migration of chronic lymphocytic leukemia (CLL) B cells and induces apoptosis. It completely inhibits heparin-induced immune complex (HIT) antibody-mediated aggregation of human platelets and transgenic HIT mouse platelets. The compound also inhibits B cell receptor signaling, reducing the expression of activation markers (e.g., CD69) and pro-survival factors (e.g., Bcl-2, Mcl-1). |
| ln Vivo |
PRT-060318 (PRT318) (10 and 30 mg/kg, by gavage, twice daily for 7 days) dihydrochloride can prevent heparin-induced acute symptoms and thrombosis in an injection model [1].
In vivo, PRT-060318 dihydrochloride prevents heparin-induced thrombocytopenia and thrombosis in a transgenic mouse model expressing human platelet factor 4. It is used to study the role of Syk in B cell malignancies, including chronic lymphocytic leukemia (CLL). The compound is orally bioavailable and shows excellent efficacy in reducing CLL tumor burden and preventing thrombosis at doses that are well tolerated. |
| Enzyme Assay |
For Syk kinase assays, recombinant Syk (10-50 ng) is incubated with PRT-060318 dihydrochloride (0.001-10 uM) in a kinase buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.01% Triton X-100) at 30degC for 15 minutes. The substrate (e.g., biotinylated peptide) and ATP (10 uM, with gamma-32P-ATP) are added. After 30 minutes, the reaction is spotted onto streptavidin-coated membranes or P81 filter paper, washed, and counted. IC₅0 is calculated as 3 nM. For platelet aggregation assays, human platelets are isolated from blood and incubated with PRT-060318 (0.01-10 uM) for 15 minutes. Aggregation is induced by HIT immune complexes or by GPVI agonists (e.g., convulxin). Aggregation is measured using a light transmission aggregometer.
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| Cell Assay |
For CLL B cell assays, primary CLL B cells are isolated from patient blood and cultured in RPMI-1640 with 10% FBS. Cells are treated with PRT-060318 dihydrochloride (0.01-10 uM) for 24-72 hours. Cell viability is measured by MTT or by staining with Annexin V/PI and flow cytometry. B cell migration is assessed using a Transwell assay with CXCL12 as the chemoattractant. B cell receptor (BCR) signaling is measured by Western blotting for phospho-Syk, phospho-AKT, and phospho-ERK. Surface activation markers (e.g., CD69) are measured by flow cytometry. For platelet assays, human platelets are isolated as described above.
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| Animal Protocol |
Animal/Disease Models: C57Bl/6 mice treated with KKO (20 mg/kg, i.p.)[1]
Doses: 10 and 30 mg/kg Route of Administration: i.g., b.i.d. for 7 days Experimental Results: Significantly inhibited platelet deposition without affecting bleeding time. Markedly reduced HIT immune complex-induced thrombosis in the lungs. Significantly reduced the thrombosis score compared to vehicle. For CLL efficacy studies, immunodeficient mice (e.g., NSG or Rag2-/-gammac-/-) are engrafted with primary human CLL B cells (5-10×10⁶ cells/mouse) via tail vein injection. After engraftment is confirmed (2-4 weeks), PRT-060318 dihydrochloride is administered orally at doses of 10-50 mg/kg in a vehicle (e.g., 0.5% methylcellulose or 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline) once or twice daily for 4-8 weeks. CLL burden is monitored by measuring human CD19+CD5+ cells in peripheral blood by flow cytometry. Splenomegaly and lymphadenopathy are assessed by palpation or by MRI. For thrombosis studies, transgenic mice expressing human PF4 are injected with heparin (e.g., 100 U/kg i.v.) followed by anti-PF4/heparin antibodies. PRT-060318 is administered orally 1 hour before heparin injection. Platelet count is measured, and thrombosis is assessed by histology of the lungs. |
| ADME/Pharmacokinetics |
PRT-060318 dihydrochloride is orally bioavailable (F% ~50-80%). After oral administration, peak plasma concentrations (Cmax) are achieved within 0.5-2 hours. The terminal elimination half-life (t1/2) is approximately 3-6 hours in rodents. The compound undergoes hepatic metabolism, primarily via CYP3A4/5, and is excreted in feces and urine. It is highly protein bound (>90%). The PK profile supports once- or twice-daily dosing in animal models and potential clinical use.
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| Toxicity/Toxicokinetics |
In preclinical studies, PRT-060318 dihydrochloride is well tolerated at therapeutic doses (10-30 mg/kg daily). No significant body weight loss or gross toxicity is observed. At higher doses (>100 mg/kg), mild hepatotoxicity (elevated ALT/AST) and gastrointestinal disturbances may occur. It is not mutagenic in the Ames test. It does not significantly inhibit hERG channels (IC50 > 10 uM), suggesting a low risk of cardiac arrhythmias. Comprehensive toxicology studies are ongoing.
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| References |
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| Additional Infomation |
PRT-060318 dihydrochloride is a potent, selective, and orally active Syk inhibitor developed for the treatment of B cell malignancies, such as chronic lymphocytic leukemia (CLL), and for preventing heparin-induced thrombocytopenia and thrombosis (HIT). It has completed preclinical studies and has been evaluated in clinical trials for the treatment of immune thrombocytopenia and rheumatoid arthritis. It is a research compound and is not yet approved for general clinical use.
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| Molecular Formula |
C18H26CL2N6O
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| Molecular Weight |
413.34
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| CAS # |
3032567-93-1
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| Related CAS # |
PRT-060318; 1194961-19-7
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| Appearance |
Light yellow to brown solid powder
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| Synonyms |
PRT318 dihydrochloride
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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, avoid exposure to moisture. |
| 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 | 2.4193 mL | 12.0966 mL | 24.1932 mL | |
| 5 mM | 0.4839 mL | 2.4193 mL | 4.8386 mL | |
| 10 mM | 0.2419 mL | 1.2097 mL | 2.4193 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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