| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg | |||
| Other Sizes |
| Targets |
PAK1 (Ki = 23.3 nM); PAK2 (Ki = 72.4 nM); PAK4 (Ki = 2.4 μM)
FRAX1036 targets Group I p21-activated kinases (PAKs), specifically PAK1, PAK2, and PAK3. PAKs are serine/threonine kinases that are downstream effectors of the small GTPases Rac and Cdc42. They play important roles in cell proliferation, migration, and survival. By inhibiting PAK1 and PAK2 with high potency, FRAX1036 blocks signaling pathways involved in tumorigenesis. |
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| ln Vitro |
FRAX1036 is a PAK inhibitor that has corresponding Kis values of 23.3 nM, 72.4 nM, and 2.4 μM for PAK1, PAK2, and PAK4. When FRAX1036 (2.5 μM) and docetaxel are combined, it changes the phosphorylation of stathmin, causes the apoptotic marker cleaved PARP, and accelerates the apoptotic process in MDA-MB-175 and HCC2911 cells.It also modifies the organization of microtubules, mitosis, and cell fate in U2OS cells. Additionally, FRAX1036 exhibits notably efficient inhibition on U2OS cells [1]. When combined with KRAS prenylation inhibitors, FRAX1036 (10 μM) inhibits the growth of non-small cell lung cancer (NSCLC) cells[2]. |
| ln Vivo |
Frax1036 treatment slows the growth of KT21 tumors and is unlikely to cause significant blood brain barrier permeability in mice.
In vivo, FRAX1036 has been shown to deter proliferation in ovarian cancer cell models. Its ability to inhibit PAK1 and PAK2 activity makes it a promising candidate for cancer therapy. It is being investigated in preclinical studies for its potential in treating cancers with PAK dysregulation. |
| Enzyme Assay |
The phosphorylation of a FRET peptide substrate (Ser/Thr19) labeled with coumarin and fluorescein is measured using the Z'-LYTETM assay to determine the activity/inhibition of human recombinant PAK1 (kinase domain), PAK2 (full length), or PAK4 (kinase domain). The samples used in the 10 μL assay mixtures include 20 pM PAK1; 50 pM PAK2; 90 pM PAK4), 0.01% Brij-35, 10 mM MgCl2, 1 mM EGTA, and 50 mM HEPES (pH 7.5). In black polypropylene 384-well plates, incubations are conducted at 22°C. The assay is started by adding 2.5 μL of assay buffer containing 4× ATP (160 μM PAK1; 480 μM PAK2; 16 μM PAK4). The enzyme, FRET peptide substrate, and serially diluted test compounds (FRAX1036, etc.) are preincubated together in assay buffer (7.5 μL) for 10 minutes prior to the assay. After the assay mixtures are incubated for 60 minutes, 5 μL of Z'-LYTETM development reagent is added to quench them, and an hour later, the emissions of fluorescein (520 nm) and coumarin (445 nm) are measured following excitation at 400 nm. A measurement is made of the emission ratio (445 nm/520 nm) to determine the extent of substrate phosphorylation[1].
In vitro kinase assays for FRAX1036 measure its inhibition of PAK1, PAK2, and other PAK isoforms. The compound is incubated with each kinase and a peptide substrate in the presence of ATP. The Ki values are determined from kinetic studies. |
| Cell Assay |
Prior to being treated with DMSO, FRAX1036, and/or docetaxel, cells are plated at 10,000 cells/well in 96-well plates for 24 hours in order to perform caspase 3/7 activation apoptosis assays. The reagent caspase 3/7 is added at a dilution of 1:1000. Using an IncuCyte Zoom Live-content imaging system, cells are imaged at 10× magnification at 37°C and 5% CO2. During the course of 36 to 72 hours, two images are acquired per well, every two or four hours. IncuCyte analysis software is used to identify and measure green (apoptotic) cells/images in data. Every condition is carried out three times. Excel is used to plot averages with SEM at each time point. When comparing the combination of FRAX1036 and docetaxel with each individual agent in Prism at the final time point, a t-test is run. The final apoptotic cell count is divided by the total cell count to determine the apoptotic index based on the apoptosis assays. Excel is used to plot averages with SEM, and a t-test is used to compare the efficacy of FRAX1036 and docetaxel in combination with each agent used alone in Prism[1]. |
| Animal Protocol |
Pak2-deficient mice
30 mg/kg by oral gavage In vivo animal experiments for FRAX1036 have been conducted in ovarian cancer models. Tumor-bearing mice are treated with the compound, and tumor growth inhibition is monitored. Its effects on PAK signaling and tumor cell proliferation are assessed. |
| ADME/Pharmacokinetics |
FRAX1036 has a molecular weight of 518.10 and a molecular formula of C28H32ClN7O. It is soluble in DMSO and ethanol but insoluble in water. Its purity is ≥98%. It is typically stored at -20°C.
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| Toxicity/Toxicokinetics |
FRAX1036 is for research use only and is not intended for human use. Its safety profile is being evaluated in preclinical studies. As a potent PAK inhibitor, its toxicity is related to its mechanism of action. Standard safety precautions should be taken when handling.
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| References | |
| Additional Infomation |
FRAX1036 is a potent and selective Group I PAK inhibitor. It has a Ki of 23.3 nM for PAK1 and 72.4 nM for PAK2. It induces apoptosis in PAK1-amplified breast cancer cells. It has been studied in ovarian cancer models. It is a research compound for studying PAK signaling in cancer.
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| Molecular Formula |
C28H32CLN7O
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|---|---|
| Molecular Weight |
518.062
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| Exact Mass |
517.235
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| Elemental Analysis |
C, 64.92; H, 6.23; Cl, 6.84; N, 18.93; O, 3.09
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| CAS # |
1432908-05-8
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| Related CAS # |
1432908-05-8
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| PubChem CID |
71557891
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| Appearance |
White to light yellow solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
669.8±65.0 °C at 760 mmHg
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| Flash Point |
358.9±34.3 °C
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| Vapour Pressure |
0.0±2.0 mmHg at 25°C
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| Index of Refraction |
1.624
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| LogP |
3.73
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
37
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| Complexity |
803
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1C(C2=CC=C(C3=NC(C)=CN=C3)C=C2Cl)=CC4=CN=C(NCCC5CCN(C)CC5)N=C4N1CC
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| InChi Key |
RYCBSFIKWACFBY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C28H32ClN7O/c1-4-36-26-21(16-32-28(34-26)31-10-7-19-8-11-35(3)12-9-19)13-23(27(36)37)22-6-5-20(14-24(22)29)25-17-30-15-18(2)33-25/h5-6,13-17,19H,4,7-12H2,1-3H3,(H,31,32,34)
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| Chemical Name |
6-[2-chloro-4-(6-methylpyrazin-2-yl)phenyl]-8-ethyl-2-[2-(1-methylpiperidin-4-yl)ethylamino]pyrido[2,3-d]pyrimidin-7-one
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| Synonyms |
FRAX 1036; FRAX-1036; FRAX1036
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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) |
DMSO: ~5.3 mg/mL (~10.2 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 10 mg/mL (19.30 mM) in 50% PEG300 +50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9303 mL | 9.6514 mL | 19.3028 mL | |
| 5 mM | 0.3861 mL | 1.9303 mL | 3.8606 mL | |
| 10 mM | 0.1930 mL | 0.9651 mL | 1.9303 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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