| 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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CNX-1351 (CNX1351; CNX 1351) is a novel, potent, covalent and selective inhibitor of PI3Kα with anticancer activity. With an IC50 of 6.8 nM, it inhibits PI3Kα . Since PI3Kβ, -γ, and -δ are not covalently modified, CNX-1351 works by covalently altering PI3Kα on cysteine 862 (C862), an amino acid specific to the α isoform. It has a strong antiproliferative effect (GI(50) < 100 nM) and is able to potently (EC(50) < 100 nM) and specifically inhibit signaling in PI3Kα-dependent cancer cell lines.
| Targets |
PI3Kα (IC50 = 6.8 nM); PI3Kβ (IC50 = 166 nM); PI3Kδ (IC50 = 240.3 nM); PI3Kγ (IC50 = 3020 nM)
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| ln Vitro |
CNX-1351 exhibits a strong anti-proliferative action (GI50<100 nM) by efficiently (EC50<100 nM) inhibiting signaling in the PI3Kα support system. CNX-1351 suppresses PI3K signaling in SKOV3 cells CNX-1351 was utilized, and cell growth was tracked, to investigate the functional effects of suppressing PI3Kα. When CNX-1351 was exposed to both PIK3CA-driven cell lines for 96 hours (GI50 78 and 55 nM, respectively), the cell lines' proliferation was restrained [1].
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| ln Vivo |
In mouse spleens, CNX-1351 binds to PI3Kα and suspends p-AktSer473. For five days in a row, CNX-1351 was hung into the abdomen cavity of three naked mice per group at a concentration of 100 mg/kg once daily. Mice's spleens were removed at predetermined intervals (1–24 hours) following the final session, and P-AktSer473 or PI3Kα occupancy was measured by immunoblotting. The last indication of PI3K arm signaling inhibition was P-AktSer473). After one hour and four hours, decrease [1].
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| Enzyme Assay |
CNX-1351 is tested in a panel of 10 lipid kinases. CNX-1351s tested in a 10-concentration IC50 curve with 3-fold serial dilution starting at 1 μM. At 10 μM ATP, reactions are carried out. For PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ, an HTRF assay format is utilized; for other kinases, an ADP-GLO assay format is utilized. Phosphatidylinositol 4,5-bisphosphate is the substrate for HTRF, sphingosine is the substrate for SPHK1 and SPHK2, and phosphatidylinositol is the substrate for all other ADP-GLO enzymes. CNX-1351 is run in a kinase selectivity panel using HotSpot technology and radioisotope-based P81 filtration for general kinase selectivity. Pure DMSO is used to dissolve CNX-1351 to a final test concentration of 1 μM. The various kinases tested against CNX-1351's substrates are given freshly prepared reaction buffers each day. Following the addition of the kinase, any necessary cofactors are added to the substrate solution, which is then preincubated for 30 minutes at room temperature. The reaction is started by adding 10 μM of 33P-ATP to the reaction mixture. The reaction then proceeds for the next two hours at room temperature. The reaction is stopped, and any phosphate that hasn't already reacted is removed using 0.1% phosphoric acid before being detected with a special technology. A non-selective, ATP-competitive kinase inhibitor called 10 M staurosporine is used as the positive control in the study, which is carried out in duplicate[1].
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| Cell Assay |
The SKOV3 proliferation assay medium (DMEM supplemented with 5-10% FBS and pen/strep) is plated with SKOV3 cells or MCF-7 cells at a density of 5000 cells in 180 μL volume per well in Costar no. 3610 white 96-well clear flat-bottom plates. The plates are then incubated overnight at 37°C in a humid incubator. After setting up a standard curve with a cell count ranging from 10,000 to 50,000 in a different plate and allowing it to adhere to the plate for 4-6 hours, Cell Titer-Glo is used to develop the plate. The following morning, 3-fold compound dilutions between 10,000 and 40 nM are made in proliferation medium with 1% DMSO. In order to create a dose-response curve from 1000 to 4 nM, 20 μL of each dilution is then added to SKOV3 or MCF-7 cells that were plated the day before. Cell Titer Glo is used to develop the cells after 96 hours of incubation. Utilizing the standard curve produced at the assay's beginning, the cell counts at the assay's conclusion are calculated. The following formulas are used to determine the GI50 values and calculate growth inhibition[1].
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| Animal Protocol |
Mice：Compound (GDC-0941 or CNX-1351) delivered ip at 100 mg/kg once daily for 5 straight days is given to female nu/nu (n=3/group). Spleens from treated animals are removed at 1, 4, and 24 hours after the last dose has been administered. Spleens are instantly put in liquid nitrogen to freeze. Samples are kept at -80°C until homogenates processing. To create homogenates, mix approximately 100 μL of spleen sample with 300 μL of cell extraction buffer, Complete protease inhibitor, and PhosStop phosphatase inhibitor in a Precellys homogenizing tube that has been kept on ice. The sample is first homogenized for 15 seconds in a Precellys 24 homogenizer, then centrifuged at 16000 g for 20 minutes at 4 °C. The supernatant is transferred to a new tube, and the BCA Assay is used to calculate the protein concentration.
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| References |
[1]. Nacht M, et al. Discovery of a potent and isoform-selective targeted covalent inhibitor of the lipid kinase PI3Kα. J Med Chem. 2013 Feb 14;56(3):712-21
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| Molecular Formula |
C30H35N7O3S
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|---|---|
| Molecular Weight |
573.71
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| Exact Mass |
573.25221
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| Elemental Analysis |
C, 62.81; H, 6.15; N, 17.09; O, 8.37; S, 5.59
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| CAS # |
1276105-89-5
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| Related CAS # |
1276105-89-5
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| Appearance |
solid powder
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| SMILES |
C/C(C)=C/C(CCC(N1CCN(CC2=CC3=NC(C4=CC=CC5=C4C=NN5)=NC(N6CCOCC6)=C3S2)CC1)=O)=O
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| InChi Key |
DLNUPKDFXMWRFP-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C30H35N7O3S/c1-20(2)16-21(38)6-7-27(39)36-10-8-35(9-11-36)19-22-17-26-28(41-22)30(37-12-14-40-15-13-37)33-29(32-26)23-4-3-5-25-24(23)18-31-34-25/h3-5,16-18H,6-15,19H2,1-2H3,(H,31,34)
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| Chemical Name |
1-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-6-methylhept-5-ene-1,4-dione
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| Synonyms |
CNX-1351; CNX 1351; CNX1351
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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: ~100 mg/mL (174.3 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.36 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 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 (4.36 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 is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7430 mL | 8.7152 mL | 17.4304 mL | |
| 5 mM | 0.3486 mL | 1.7430 mL | 3.4861 mL | |
| 10 mM | 0.1743 mL | 0.8715 mL | 1.7430 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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