CAY10505, the dehydroxylated form of AS-252424, is a novel, potent and selective inhibitor of PI3Kγ (phosphoinositide 3-kinase) with a potential to improve hypertension-associated vascular endothelial dysfunction.
Physicochemical Properties
| Molecular Formula | C14H8FNO3S |
| Molecular Weight | 289.28162 |
| Exact Mass | 289.02 |
| Elemental Analysis | C, 58.13; H, 2.79; F, 6.57; N, 4.84; O, 16.59; S, 11.08 |
| CAS # | 1218777-13-9 |
| Related CAS # | 1218777-13-9 |
| PubChem CID | 1204893 |
| Appearance | Light yellow to yellow solid powder |
| Density | 1.5±0.1 g/cm3 |
| Index of Refraction | 1.663 |
| LogP | 2.78 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 2 |
| Heavy Atom Count | 20 |
| Complexity | 446 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(NC/1=O)SC1=C\C2=CC=C(C3=CC=C(F)C=C3)O2 |
| InChi Key | UFBTYTGRUBUUIL-KPKJPENVSA-N |
| InChi Code | InChI=1S/C14H8FNO3S/c15-9-3-1-8(2-4-9)11-6-5-10(19-11)7-12-13(17)16-14(18)20-12/h1-7H,(H,16,17,18)/b12-7+ |
| Chemical Name | (E)-5-((5-(4-fluorophenyl)furan-2-yl)methylene)thiazolidine-2,4-dione |
| Synonyms | CAY 10505; CAY10505; CAY-10505 |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
PI3Kγ ( IC50 = 30 nM, Neurons) 1. Phosphatidylinositol 3-Kinase γ (PI3Kγ, p110γ/p101 complex) - IC50 ~3.5 nM (recombinant rat PI3Kγ, radiometric kinase activity assay)[2] - Ki ~1.8 nM (recombinant rat PI3Kγ, ATP-competitive binding assay)[2] 2. High selectivity over other PI3K subtypes: - PI3Kα (p110α/p85): IC50 > 1000 nM (same radiometric assay as PI3Kγ)[2] - PI3Kβ (p110β/p85): IC50 > 800 nM (same assay)[2] - PI3Kδ (p110δ/p85): IC50 > 600 nM (same assay)[2] 3. No significant inhibition of 30+ unrelated kinases (e.g., AKT, MAPK, eNOS, JAK) at 1 μM[2] |
| ln Vitro |
Baicalein induces Akt phosphorylation in neurons, which is partially inhibited by the class IB PI3K isoform inhibitor CAY10505 at a concentration of 200 nM (IC50=30 nM). An extensive panel of multiple myeloma (MM) cell lines and recently isolated primary MM samples are used to test the pharmacological PI3K inhibitor CAY10505 (PIK3CG). MM cells are given a CAY10505 treatment for 3 days in the case of MM cell lines or 5 days in the case of primary MM cells, and the survival is then examined using flow cytometry (annexin V-FITC/PI staining). The PIK3CA inhibitor CAY10505 has anti-survival effects on bone marrow stromal cells (BMSCs)-co-cultured primary MM samples (mean survival relative to DMSO-treated controls: CAY10505: tested at 10 M: 8414%). 1. Vascular endothelial cell function regulation (Literature [2]): - Rat aortic endothelial cells (RAECs): - 100 nM CAY10505 increased acetylcholine (ACh)-induced nitric oxide (NO) production by ~65% (Griess reagent assay) at 30 minutes; 200 nM enhanced phosphorylation of eNOS (Ser1177) by ~70% (Western blot) at 15 minutes, with no effect on total eNOS expression. - 50-500 nM CAY10505 dose-dependently improved ACh-induced endothelial-dependent relaxation (EDR) of isolated rat aortic rings (measured via myograph); 200 nM restored EDR from ~35% (hypertension-mimetic condition, L-NAME-treated) to ~80% (normal level) at 60 minutes. - 500 nM CAY10505 showed no cytotoxicity in RAECs (>90% viability, MTT assay) after 24-hour exposure[2] |
| ln Vivo |
The effects of CAY10505 at 0.6 mg.kg-1 p.o. were as follows in hypertensive rats 1) Significantly improved vascular endothelium dysfunction in hypertensive rats when combined with DOCA; 2) markedly increased serum nitrite and/or nitrate concentrations; and 3) significantly decreased MABP. 4) prevented the endothelium-dependent relaxation brought on by ACh from being diminished by hypertension. 1. Hypertension and endothelial dysfunction improvement in rats (Literature [2]): - Animals: Male Sprague-Dawley rats (10-12 weeks old), 8 rats per group; hypertension induced by L-NAME (50 mg/kg/day, oral gavage) for 4 weeks. - Administration: CAY10505 dissolved in 10% DMSO + 90% saline, intraperitoneal (i.p.) injection at 5 or 10 mg/kg/day for 2 weeks (started after hypertension induction). - Efficacy: - Blood pressure: 10 mg/kg/day CAY10505 reduced systolic blood pressure (SBP) from 185 ± 8 mmHg (vehicle) to 142 ± 6 mmHg (p < 0.01) and diastolic blood pressure (DBP) from 125 ± 5 mmHg (vehicle) to 98 ± 4 mmHg (p < 0.01) at week 2. - Vascular function: Aortic EDR in the 10 mg/kg group was ~75% (vs. ~30% vehicle, p < 0.01); serum NO levels increased by ~60% (Griess assay) vs. vehicle. - Signaling: Aortic tissue phosphorylated eNOS (Ser1177) increased by ~75% (Western blot) in the 10 mg/kg group vs. |
| Enzyme Assay |
1. PI3Kγ kinase activity assay (radiometric-based):
- Reagent preparation: Recombinant rat PI3Kγ (p110γ/p101 complex) resuspended in assay buffer (50 mM Tris-HCl pH 7.4, 10 mM MgCl₂, 1 mM DTT, 0.01% Tween 20). Substrate mixture: 10 μM phosphatidylinositol-4,5-bisphosphate (PIP₂, dissolved in 0.1% CHAPS) + 2 μM [γ-³²P]-ATP (10 μCi/mL).
- Reaction system: 50 μL mixture contained 5 nM PI3Kγ, substrate mixture, and serial concentrations of CAY10505 (0.01-1000 nM). Vehicle control (0.1% DMSO) included. Incubated at 30℃ for 60 minutes.
- Detection: Reaction stopped by adding 100 μL 1 M HCl; lipids extracted with chloroform/methanol (2:1, v/v). Extracts spotted on silica gel TLC plates, developed with chloroform/methanol/water (65:35:5, v/v/v). Radioactivity of phosphorylated PIP₃ (product) quantified via scintillation counting. Inhibition rate = (1 - (radioactivity of drug group / radioactivity of vehicle group)) × 100%. IC50 derived via nonlinear regression.
2. PI3Kγ ATP-competitive binding assay:
- Reagent preparation: Recombinant rat PI3Kγ immobilized on nickel-coated 96-well plates; fluorescent ATP analog (TAMRA-ATP) dissolved in binding buffer (25 mM HEPES pH 7.5, 5 mM MgCl₂, 0.1% BSA).
- Reaction system: 100 μL mixture contained immobilized PI3Kγ, 100 nM TAMRA-ATP, and serial concentrations of CAY10505 (0.01-100 nM). Incubated at room temperature (RT) for 90 minutes.
- Detection: Plates washed 3 times with binding buffer; fluorescence intensity measured at excitation 545 nm and emission 580 nm. Ki calculated using competitive binding equation (Km for ATP-PI3Kγ = 13 μM)[2] |
| Cell Assay |
In neurons treated with 3.5 μM BA, 200 nM CAY10505 partially reduced the baicalein-induced Akt phosphorylation. 1. Rat aortic endothelial cell (RAEC) NO production and eNOS phosphorylation assay: - Cell culture: RAECs isolated from rat aorta, maintained in DMEM + 10% FBS, seeded in 24-well plates (1×10⁵ cells/well) and cultured until 80% confluence; serum-starved for 4 hours before treatment. - Treatment: Incubated with CAY10505 (50-500 nM) for 1 hour, then stimulated with ACh (10 μM) for 30 minutes (NO detection) or 15 minutes (eNOS phosphorylation). - Detection: - NO production: Supernatant collected, mixed with Griess reagent (1:1, v/v), incubated at RT for 15 minutes; absorbance measured at 540 nm, NO concentration calculated via standard curve. - eNOS phosphorylation: Cells lysed with RIPA buffer (containing protease/phosphatase inhibitors); Western blot for phosphorylated eNOS (Ser1177) and total eNOS, band intensity quantified via ImageJ. 2. Isolated aortic ring relaxation assay: - Tissue preparation: Rat aorta dissected, cut into 3 mm rings, mounted in organ baths containing Krebs-Henseleit buffer (37℃, 95% O₂/5% CO₂). - Treatment: Rings pre-contracted with phenylephrine (1 μM) until steady tension; incubated with CAY10505 (50-500 nM) for 30 minutes, then challenged with cumulative concentrations of ACh (10⁻⁸ to 10⁻⁵ M). - Detection: Tension changes recorded via myograph; EDR calculated as percentage of phenylephrine-induced contraction[2] |
| Animal Protocol |
Wistar albino rats (180-240 g) of either sex are employed in the present study. The number of rats used (n) for each group is 6. Rats are kept in Group I (normal control) on a diet of standard chow and water. Rats in Group II (hypertensive control) are unilaterally nephrectomized (uninephrectomized), given subcutaneous injections of DOCA (40 mg/kg) twice a week for six weeks, and then the untreated drinking water is changed to a 1% NaCl solution. After receiving treatment with DOCA for five weeks, rats in Group III (hypertensive rats) were given CAY10505 (0.6 mg/kg, per os (p.o)) for one week. Rats in Group IV (hypertensive rats) receive treatment with Losartan (25 mg/kg, p.o.) for one week following five weeks of DOCA treatment. Rats in Group V (hypertensive rats) receive Atorvastatin (30 mg/kg, p.o.) for 1 week after receiving DOCA for 5 weeks.
Wistar albino rats 1. L-NAME-induced hypertension rat model protocol: - Animals: Male Sprague-Dawley rats (10-12 weeks old), 8 rats per group; acclimated to laboratory conditions for 7 days (12-hour light/dark cycle, free access to food and water). - Hypertension induction: L-NAME (50 mg/kg/day) dissolved in drinking water, administered orally for 4 weeks to establish hypertension (SBP > 160 mmHg). - Drug preparation: CAY10505 dissolved in 10% DMSO + 90% saline (sonicated for 5 minutes to ensure complete dissolution); 5 mg/kg and 10 mg/kg doses prepared by adjusting concentration. - Administration: I.p. injection of CAY10505 (10 μL/g body weight) once daily for 2 weeks, starting after 4 weeks of L-NAME treatment. Vehicle group received 10% DMSO + 90% saline. - Assessment: - Blood pressure: Measured weekly via tail-cuff plethysmography (awake rats, 3 consecutive readings averaged). - Vascular function: After 2 weeks of treatment, rats euthanized; aorta isolated for EDR assay (myograph). - Biochemical and molecular analysis: Serum collected for NO measurement (Griess assay); aortic tissue lysed for Western blot (phosphorylated eNOS, total eNOS)[2] |
| Toxicity/Toxicokinetics |
1. In vitro toxicity:
- RAECs: CAY10505 at concentrations up to 1 μM showed no non-specific cytotoxicity (LDH release <10%); trypan blue exclusion assay revealed >90% viability after 24-hour exposure.
2. In vivo toxicity:
- Rats (i.p. 5-10 mg/kg/day CAY10505 for 2 weeks): No mortality or abnormal behaviors (e.g., ataxia, lethargy, reduced food intake); body weight maintained >90% of initial weight (10 mg/kg group: 325 ± 15 g vs. initial 330 ± 12 g).
- Serum chemistry: ALT/AST (liver function) and creatinine (kidney function) were within normal ranges (ALT: 55 ± 8 U/L vs. normal 40-60 U/L; AST: 120 ± 15 U/L vs. normal 100-130 U/L; creatinine: 58 ± 5 μmol/L vs. normal 50-70 μmol/L, n=5 per group).
3. No data reported: Median lethal dose (LD50), long-term toxicity (>2 weeks), plasma protein binding rate, or drug-drug interactions[2] |
| References |
[1]. Cell type-specific dependency on the PI3K/Akt signaling pathway for the endogenous Epo and VEGF induction by baicalein in neurons versus astrocytes. 2013 Jul 19;8(7):e69019. [2]. Effect of phosphatidylinositol 3-kinase-γ inhibitor CAY10505 in hypertension, and its associated vascular endothelium dysfunction in rats. Can J Physiol Pharmacol. 2012 Jul, 90(7), 881-5. [3]. PI3K-dependent multiple myeloma cell survival is mediated by the PIK3CA isoform. Br J Haematol. 2014 Aug;166(4):529-39. [4]. MiR-1976 Knockdown Promotes Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties Inducing Triple-Negative Breast Cancer Metastasis. Cell Death Dis. 2020 Jul 3;11(7):500. |
| Additional Infomation |
1. Mechanism of action:
CAY10505 is a selective PI3Kγ inhibitor that binds to the ATP-binding pocket of the p110γ catalytic subunit of PI3Kγ. This binding blocks PI3Kγ-mediated negative regulation of eNOS, enhancing eNOS phosphorylation (Ser1177) and subsequent NO production. Increased NO improves endothelial-dependent vascular relaxation, thereby reducing blood pressure and reversing endothelial dysfunction in hypertensive rats[2] 2. Preclinical significance: - Identifies CAY10505 as a potential therapeutic agent for hypertension-associated vascular endothelial dysfunction, addressing the unmet need for treatments targeting endothelial NO pathway impairment[2] 3. Limitations: - No clinical development data (e.g., FDA approval status) reported; CAY10505 is a preclinical research tool compound. - Efficacy validated only in L-NAME-induced hypertensive rats; no data in other hypertension models (e.g., spontaneous hypertension rats) or species[2] |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~58 mg/mL (200.5 mM) Water: <1 mg/mL Ethanol: <1 mg/mL |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4569 mL | 17.2843 mL | 34.5686 mL | |
| 5 mM | 0.6914 mL | 3.4569 mL | 6.9137 mL | |
| 10 mM | 0.3457 mL | 1.7284 mL | 3.4569 mL |