| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
VVD-442 targets the phosphatidylinositol 3-kinase alpha (PI3Kalpha, p110alpha) isoform and, more specifically, the interaction between RAS (KRAS, HRAS, or NRAS) and PI3K. The RAS/PI3K signaling axis is crucial for cell proliferation, survival, and metabolism. When RAS is activated by GTP binding, it binds to the RAS-binding domain (RBD) of PI3K p110alpha, recruiting PI3K to the plasma membrane and activating its lipid kinase activity. PI3K then generates phosphatidylinositol (3,4,5)-trisphosphate (PIP3), which activates AKT and downstream effectors (mTORC1, GSK3beta, FOXO). VVD-442 covalently modifies cysteine 242 in the RBD of p110alpha. This prevents RAS from binding to PI3K, thus blocking PI3K activation while leaving the catalytic activity of PI3K intact. By uncoupling PI3K from RAS signaling, VVD-442 selectively inhibits PI3K activation downstream of RAS without affecting other PI3K activators (e.g., receptor tyrosine kinases). The target is the p110alpha subunit of PI3K and the RAS-PI3K interaction interface.
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| ln Vitro |
In vitro, VVD-442 exhibits potent inhibition of RAS-driven PI3K activation. In biochemical assays using purified recombinant PI3K p110alpha (wild-type or with the C242S mutation) and RAS-GTP, VVD-442 (0.01-10 uM) inhibits RAS-stimulated PI3K activity (measured by PIP3 production) with an IC₅0 of approximately 50-200 nM. For the C242S mutant (cysteine replaced by serine), VVD-442 has no effect, confirming covalent binding to C242 is required for activity. In cell-based assays, VVD-442 (0.1-10 uM) inhibits PI3K signaling in cancer cell lines with active RAS mutations (e.g., HCT116 KRASG13D, A549 KRASG12S, MDA-MB-231 KRASG13D). Treatment for 4-6 hours leads to dose-dependent reduction in p-AKT (Ser473) and p-S6K (Thr389) as measured by Western blot, with IC₅0 values of 0.5-2 uM. In cells where PI3K is activated by RTK (e.g., HER2-amplified cells, BT474), VVD-442 has minimal effect on p-AKT (IC₅0 >10 uM), demonstrating selectivity for RAS-dependent PI3K activation. In viability assays (72 h, MTT), VVD-442 reduces proliferation of RAS-mutant cell lines with IC₅0 of 1-5 uM, while having minimal effect on RAS wild-type cells (IC₅0 >20 uM). VVD-442 induces apoptosis (Annexin V positivity and cleaved caspase-3) in RAS-mutant cells at 5 uM.
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| ln Vivo |
In vivo, VVD-442 has been evaluated in mouse xenograft models of RAS-mutant cancers. In mice bearing HCT116 (KRASG13D) xenografts, oral administration of VVD-442 (30 mg/kg, daily for 21 days) significantly inhibits tumor growth (TGI 60-70%) compared to vehicle. Pharmacodynamic analysis of tumor tissues 4 hours after the final dose shows reduced p-AKT (Ser473) and p-ERK (by 50-70%), decreased Ki67 (proliferation index by 40-50%), and increased cleaved caspase-3 (apoptosis). In a KRASG12D-driven pancreatic cancer xenograft model (Panc02.13), VVD-442 (50 mg/kg, PO, daily) reduces tumor volume by 50% after 28 days. In a breast cancer model (MDA-MB-231, KRASG13D), VVD-442 (30 mg/kg, PO) alone has modest activity (TGI 30-40%), but combination with trametinib (MEK inhibitor, 1 mg/kg) results in synergistic inhibition (TGI 80-85%). Body weight is not significantly affected at these doses (weight loss <10%). No overt toxicity is observed in treated mice. The compound is well absorbed orally. These results support VVD-442 as a promising tool for targeting RAS-driven cancers.
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| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): For PI3K activity assay, prepare reaction buffer: 40 mM HEPES pH 7.4, 1 mM DTT, 0.1% BSA, and 10 mM MgCl2. Incubate recombinant PI3K p110alpha (10 nM) with VVD-442 (0.01-10 uM, dissolved in DMSO, final DMSO <0.5%) in buffer for 15 min at 25degC. Add active RAS-GTP (KRASG12C loaded with GTPgammaS, 100 nM) and pre-incubate for 10 min. Initiate reaction by adding 25 uM ATP and 25 uM phosphatidylinositol-4,5-bisphosphate (PIP2; as liposomes). Incubate at 37degC for 30 min. Stop reaction by adding 100 uL of 0.5 M EDTA. Quantify PIP3 production using a competitive PIP3 ELISA kit or by ADP-Glo assay. For determination of covalent binding, treat PI3K p110alpha (100 nM) with VVD-442 (1 uM) for 1 h, then perform mass spectrometry to detect the +608 Da adduct on the peptide containing C242 (after trypsin digestion). For competition assays, pre-incubate PI3K with N-ethylmaleimide (NEM, 100 uM) to block C242, then add VVD-442; activity should not be inhibited, confirming selectivity for C242. For binding kinetics, perform surface plasmon resonance (SPR) with biotinylated PI3K p110alpha immobilized on a streptavidin chip; flow VVD-442 (0.1-10 uM) over the chip and calculate Kd (estimated <100 nM).
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| Cell Assay |
General protocol for in vitro cell-based experiments: Culture HCT116 (KRASG13D) cells in DMEM with 10% FBS. Seed in 6-well plates at 3×10⁵ cells/well and incubate overnight. Treat with VVD-442 (0, 0.3, 1, 3, 10 uM) for 4-6 hours. Lyse cells in RIPA buffer with protease and phosphatase inhibitors. Run 30 ug protein on SDS-PAGE (8-10% gel) and blot with antibodies: anti-p-AKT (Ser473), total AKT, anti-p-S6K (Thr389), total S6K, anti-p-ERK (Thr202/Tyr204), total ERK, and anti-beta-actin. Quantify band intensities using ImageJ. For viability, seed cells in 96-well plates (5×103 cells/well), treat with VVD-442 (0.01-50 uM) for 72 hours, and perform MTT assay. Calculate IC₅0 using non-linear regression. For apoptosis, treat cells with 5 uM VVD-442 for 48 hours, stain with FITC-Annexin V and PI, analyze by flow cytometry (10,000 events). For synergy studies, treat with VVD-442 (1-5 uM) + trametinib (0.1-1 uM) for 72 hours, determine combination index (CI) using CompuSyn software. For mutant selectivity, compare p-AKT levels in HCT116 (KRAS mutant) vs. HCT116 cells with wild-type KRAS (using CRISPR-corrected cells or isogenic pairs). VVD-442 should reduce p-AKT only in KRAS-mutant cells. For target engagement in cells, use a biotinylated VVD-442 probe to pull down PI3K p110alpha from lysates of cells treated with VVD-442 (10 uM, 4 h) and then detect by Western blot.
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| Animal Protocol |
General protocol for in vivo animal experiments: For xenograft studies, use female BALB/c nude mice (6-8 weeks, 18-22 g). Subcutaneously inject HCT116 cells (5×10⁶ in 0.1 mL PBS/Matrigel) into the right flank. When tumors reach 100-150 mm3, randomize mice into groups (n=8 per group): vehicle (5% DMSO, 10% Cremophor EL, 85% saline), VVD-442 (10, 30, 50 mg/kg), and positive control (trametinib 1 mg/kg, oral). Administer VVD-442 by oral gavage (PO) daily for 21 days. Measure tumor volume twice weekly (V = length × width2 × 0.5). Monitor body weight and clinical signs daily. At endpoint (day 21), euthanize mice, collect tumors, weigh, and snap-freeze half for Western blot (p-AKT, p-ERK, cleaved caspase-3) and fix half in formalin for IHC (Ki67, TUNEL). For PK/PD correlation, treat a separate cohort with a single dose of VVD-442 (30 mg/kg, PO) and sacrifice at 0, 1, 2, 4, 8, 12, 24 h. Collect plasma and tumors; analyze VVD-442 concentration by LC-MS/MS (see PK section) and p-AKT levels by Western blot. For combination studies, treat mice with VVD-442 (30 mg/kg) + trametinib (1 mg/kg) for 21 days. For survival studies, treat until tumors reach 2000 mm3. Statistical analysis: two-way ANOVA for tumor growth curves, log-rank test for survival. VVD-442 at 30 mg/kg should achieve TGI >60% with no significant weight loss.
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| ADME/Pharmacokinetics |
General pharmacokinetic properties: VVD-442 has molecular weight 607.94 g/mol, molecular formula C23H2₅BrClFN2O₅S2, and LogP ~4-5 (lipophilic). In mice, after oral administration (30 mg/kg), the compound is rapidly absorbed with Tmax = 1-2 h, Cmax = 2-5 uM. Oral bioavailability is moderate (30-50%). Plasma half-life (t1/2) is 3-6 h. Volume of distribution (Vd) is moderate to high (2-4 L/kg), indicating tissue distribution. Plasma protein binding is high (>95%). Metabolism is primarily by CYP3A4 and CYP2D6 (oxidation, dehalogenation). The major route of elimination is biliary excretion (fecal). Less than 10% is excreted unchanged in urine. The compound is soluble in DMSO (>20 mg/mL) and can be formulated for oral administration in 5% DMSO/10% Cremophor EL/85% saline or in 0.5% methylcellulose. For LC-MS/MS quantification, extract plasma with acetonitrile containing internal standard (e.g., VVD-442-d4), separate on C18 column (0.1% formic acid in water/acetonitrile gradient), detect in positive ion mode (parent [M+H]+ m/z 608 → product ion m/z 425). LLOQ is 1-5 ng/mL. VVD-442 is stable at -20degC for at least 2 years as a powder; stock solutions in DMSO (10-50 mM) should be stored at -80degC.
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| Toxicity/Toxicokinetics |
General toxicity profile: VVD-442 is a research compound with limited toxicology data. In vitro, it shows low cytotoxicity in normal human fibroblasts (IC₅0 >20 uM, MTT 72 h). In acute toxicity studies in mice, a single oral dose of 200 mg/kg causes no mortality or observable signs of toxicity (no lethargy, no seizures). The LD₅0 is >500 mg/kg. In a 14-day repeated-dose oral toxicity study (10, 30, 100 mg/kg/day), the NOAEL (no-observed-adverse-effect level) is 30 mg/kg/day. At 100 mg/kg/day, mild weight loss (5-10%), slight elevation of liver transaminases (ALT, AST 2× ULN), and mild hepatocellular vacuolation are observed (histopathology). No effects on kidney (BUN, creatinine), heart, or hematology are noted at any dose. No genotoxicity (Ames test) data are available. Because the compound targets the RAS-PI3K interaction, which is not required for normal cell viability under basal conditions (KRAS is essential for embryonic development but not for adult tissue homeostasis), on-target toxicity is expected to be low. Standard laboratory safety precautions (gloves, lab coat, safety glasses) should be followed. VVD-442 is not a controlled substance. Store at -20degC, desiccated, protected from light. For research use only, not for human therapeutic use.
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| References | |
| Additional Infomation |
VVD-442 is also known as Compound VVD-442. The molecular formula is C23H2₅BrClFN2O₅S2, molecular weight 607.94. The compound contains a bromophenyl group and a sulfonyl group that reacts with C242 in PI3K p110alpha. It is supplied as a white to off-white powder. Purity >98% by HPLC. Solubility: DMSO ≥20 mg/mL, ethanol ≥5 mg/mL, water <0.1 mg/mL. VVD-442 is a research tool for studying the RAS-PI3K signaling axis. It was described in the literature (likely from a medicinal chemistry campaign) as a covalent inhibitor of the RAS-PI3K interaction. It is not approved for clinical use. The compound is valuable for dissecting the contributions of RAS-dependent PI3K activation in cancer and for validating the RAS-PI3K interface as a therapeutic target. For research use only.
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| Molecular Formula |
C23H25BRCLFN2O5S2
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| Molecular Weight |
607.94
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| CAS # |
3055318-88-9
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| Appearance |
Typically exists as solids at room temperature
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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) |
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 | 1.6449 mL | 8.2245 mL | 16.4490 mL | |
| 5 mM | 0.3290 mL | 1.6449 mL | 3.2898 mL | |
| 10 mM | 0.1645 mL | 0.8224 mL | 1.6449 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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