| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
PKD1 1 nM (IC50) PKD2 2.5 nM (IC50) PKD3 2 nM (IC50) PIM2 135.7 nM (IC50)
PKD1, PKD2, and PKD3 (protein kinase D isoforms), which are serine/threonine kinases involved in diverse cellular processes including cell proliferation, migration, invasion, and vesicular trafficking. |
|---|---|
| ln Vitro |
CRT0066101 (5 µM; 1 h) blocks basal and NT-induced pS916-PKD1/2 (activated PKD1/2) in Panc-1 and Panc-28 cells. CRT0066101 abolishes NT-induced Hsp27 (pS82-Hsp27) phosphorylation, attenuates PKD1-mediated NF-κB activation, and eliminates the expression of NF-κB-dependent proliferative and pro-survival proteins[1]. CRT0066101 significantly inhibits Panc-1 cell proliferation with an IC50 value of 1 µM. CRT0066101 induces apoptosis of Panc-1 cells 6-10-fold. CRT0066101 significantly reduces the proliferation of Colo357, Panc-1, MiaPaCa-2, and AsPC-1 cells, but has some effects on Capan-2 cells[1].
In cell‑free enzymatic assays, CRT0066101 inhibits PKD1 with an IC50 of 1 nM, PKD2 with an IC50 of 2.5 nM, and PKD3 with an IC50 of 2 nM. It inhibits the proliferation of PANC‑1 pancreatic cancer cells and blocks the migration and invasion of U87MG glioblastoma cells. In addition, it also inhibits PIM2 with an IC50 of approximately 135.7 nM. |
| ln Vivo |
CRT0066101 (80 mg/kg/day; oral gavage; once daily; for 21 days) effectively blocked tumor growth in vivo in the Panc-1 orthotopic model [1].
In vivo, CRT0066101 exhibits anti‑tumor activity in xenograft models of pancreatic cancer and glioblastoma. As an orally active compound, it can be administered via oral gavage to mice bearing PANC‑1 or U87MG xenografts, leading to significant inhibition of tumor growth. It also reduces metastasis in orthotopic models. |
| Enzyme Assay |
A general cell‑free protocol for PKD inhibition: A radiometric or time‑resolved fluorescence resonance energy transfer (TR‑FRET) kinase assay is used. Recombinant human PKD1 (10 nM) is incubated with a peptide substrate and 10 uM ATP in kinase buffer. Varying concentrations of CRT0066101 (0.001‑1000 nM) are added. The reaction is incubated at 30degC for 30 minutes. The amount of phosphorylated product is measured, and the IC50 is calculated. Similar assays are performed for PKD2 and PKD3.
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: Panc-1 and Panc-28 cells stimulation with neurotensin (NT) Concentration: 5 µM Incubation Duration: 1 h Experimental Results: Blocked both the basal and NT-induced pS916-PKD1/2 (activated PKD1/2). A general cellular protocol for CRT0066101: PANC‑1 pancreatic cancer cells or U87MG glioblastoma cells are seeded in 96‑well plates. Cells are treated with serial dilutions of CRT0066101 (0.01 nM to 10 uM) for 72 hours. Cell viability is measured using CellTiter‑Glo or MTT assay. For migration and invasion assays, Transwell chambers (8 um pores) are used, with or without Matrigel coating. Cells are treated with CRT0066101 (0.1‑1 uM) for 24 hours, and migrated/invaded cells are quantified. Western blot is performed for PKD downstream targets (e.g., p‑PKD, p‑CREB). |
| Animal Protocol |
Animal/Disease Models:CR-UK nu/nu mice injected with Panc-1 cells[1]
Doses: 80 mg/kg/day Route of Administration: Oral gavage; once daily; for 21 days Experimental Results: Potently blocked tumor growth in vivo. A general animal protocol for CRT0066101: Female NOD/SCID mice are subcutaneously implanted with 5×10⁶ PANC‑1 cells or U87MG cells. When tumors reach ~150 mm3, mice are randomized into treatment groups (n=8/group). CRT0066101 is formulated in a suitable vehicle (e.g., 0.5% methylcellulose or 10% DMSO, 40% PEG300, 5% Tween‑80, 45% saline) and administered via oral gavage at doses of 10, 30, and 100 mg/kg once daily for 21‑28 days. Tumor volume is measured twice weekly. At the end of the study, tumors are harvested for IHC (Ki‑67, cleaved caspase‑3), Western blot (p‑PKD, p‑CREB), and TUNEL staining. |
| ADME/Pharmacokinetics |
General PK protocol for CRT0066101: Male Sprague‑Dawley rats are administered CRT0066101 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg). Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours. Plasma concentrations are quantified by LC‑MS/MS. PK parameters (Cmax, Tmax, AUC, t1/2, clearance, Vd, and oral bioavailability) are calculated.
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| Toxicity/Toxicokinetics |
General toxicity protocol for CRT0066101: A 14‑day repeated‑dose oral toxicity study is performed in ICR mice. CRT0066101 is administered at doses of 10, 30, and 100 mg/kg/day. Parameters include clinical signs, body weight, food consumption, hematology (CBC), serum chemistry (ALT, AST, BUN, creatinine), and histopathology of major organs (liver, kidney, spleen, heart, lung, pancreas).
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| References |
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| Additional Infomation |
Protein kinase D inhibitors with anti-tumor activity
CRT0066101 has a molecular formula of C18H22N6O and a molecular weight of 338.41 g/mol. The IUPAC name is not provided. The compound is a potent and selective PKD inhibitor that has been shown to block the growth of pancreatic cancer and glioblastoma in preclinical models. |
| Molecular Formula |
C18H22N6O
|
|---|---|
| Molecular Weight |
338.41
|
| Exact Mass |
374.162
|
| CAS # |
956123-34-5
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| PubChem CID |
136189563
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| Appearance |
Solid powder
|
| LogP |
2.522
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| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
6
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| Heavy Atom Count |
25
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| Complexity |
411
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
Cl.OC1=CC=C(C2C=NN(C)C=2)C=C1C1=NC=CC(=N1)NC[C@@H](CC)N
|
| InChi Key |
SCJXQZZYGYLKJG-CQSZACIVSA-N
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| InChi Code |
InChI=1S/C18H22N6O/c1-3-14(19)10-21-17-6-7-20-18(23-17)15-8-12(4-5-16(15)25)13-9-22-24(2)11-13/h4-9,11,14,25H,3,10,19H2,1-2H3,(H,20,21,23)/t14-/m1/s1
|
| Chemical Name |
2-[4-[[(2R)-2-aminobutyl]amino]pyrimidin-2-yl]-4-(1-methylpyrazol-4-yl)phenol
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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)
|
| Solubility (In Vitro) |
DMSO : 100 mg/mL (295.50 mM; with sonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.39 mM)(Saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix well; then add 50 μL Tween-80 to the above system and mix well; then add 450 μL saline to make it 1 mL. *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.39 mM)(Saturation unknown) in 10% DMSO 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD in saline and mix well. *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. Solubility in Formulation 3: ≥ 2.5 mg/mL (7.39 mM)(saturation unknown) in 10% DMSO 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9550 mL | 14.7750 mL | 29.5500 mL | |
| 5 mM | 0.5910 mL | 2.9550 mL | 5.9100 mL | |
| 10 mM | 0.2955 mL | 1.4775 mL | 2.9550 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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