TG100713 is a novel and potent pan-PI3K (phosphatidylinositol 3-kinase) inhibitor with potential anti-inflammatory activity. It inhibits PI3Kγ, PI3Kδ, PI3Kα and PI3Kβ with respective IC50 values of 50 nM, 24 nM, 165 nM, and 215 nM. Pro-inflammatory PI3K isoformsPI3Kγ and PI3Kδ are involved in the inflammation that results in damage to ischemia-damaged tissue after reperfusion. The pro-cell survival functions of phosphoinositide 3-kinases (PI3Ks) are beneficial during tissue iscg the reperfusion phase of hemorrhage. As a result, it's possible that targeting only pro-inflammatory PI3K isoforms with durinase will help to limit the overall tissue damage brought on by ischemia/reperfusion injuries like myocardial infarction.
Physicochemical Properties
| Molecular Formula | C12H10N6O | |
| Molecular Weight | 254.2474 | |
| Exact Mass | 254.091 | |
| Elemental Analysis | C, 56.69; H, 3.96; N, 33.05; O, 6.29 | |
| CAS # | 925705-73-3 | |
| Related CAS # |
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| PubChem CID | 17751063 | |
| Appearance | Light yellow to khaki solid powder | |
| Density | 1.5±0.1 g/cm3 | |
| Boiling Point | 646.4±65.0 °C at 760 mmHg | |
| Flash Point | 344.8±34.3 °C | |
| Vapour Pressure | 0.0±2.0 mmHg at 25°C | |
| Index of Refraction | 1.815 | |
| LogP | 0.5 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 1 | |
| Heavy Atom Count | 19 | |
| Complexity | 317 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | OC1C=C(C2C=NC3N=C(N)N=C(C=3N=2)N)C=CC=1 |
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| InChi Key | UOORQSPLBHUQDQ-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C12H10N6O/c13-10-9-11(18-12(14)17-10)15-5-8(16-9)6-2-1-3-7(19)4-6/h1-5,19H,(H4,13,14,15,17,18) | |
| Chemical Name | 3-(2,4-diaminopteridin-6-yl)phenol | |
| Synonyms | TG100713; TG-100713; TG 100713 | |
| 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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
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| 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 |
PI3Kδ (IC50 = 24 nM); PI3Kγ (IC50 = 50 nM); PI3Kα (IC50 = 165 nM); PI3Kβ (IC50 = 215 nM) 1. Phosphatidylinositol 3-Kinase γ (PI3Kγ) and δ (PI3Kδ) - PI3Kγ (p110γ/p101 complex): IC50 ~10 nM (recombinant human PI3Kγ, HTRF-based kinase activity assay)[1] - PI3Kδ (p110δ/p85α complex): IC50 ~15 nM (recombinant human PI3Kδ, same HTRF assay)[1] 2. Low activity against other PI3K subtypes: - PI3Kα (p110α/p85α): IC50 > 1000 nM (same HTRF assay as PI3Kγ/δ)[1] - PI3Kβ (p110β/p85α): IC50 > 800 nM (same assay)[1] 3. No significant inhibition of 35+ unrelated kinases (e.g., AKT, ERK, JAK, eNOS) at 1 μM concentration[1] [1] |
| ln Vitro |
TG100713 (10 μM; 48 or 72 h) strongly inhibits endothelial cell (EC) proliferation[1]. 1. Inflammatory cell function inhibition (Literature [1]): - Mouse bone marrow-derived neutrophils (BMDNs): - 100 nM TG100713 inhibited fMLP-induced chemotaxis by ~70% (Transwell assay) at 2 hours; 50 nM reduced fMLP-induced actin polymerization by ~65% (phalloidin staining) at 15 minutes, a key step in cell migration. - 200 nM TG100713 reduced LPS-induced TNF-α secretion by ~80% and IL-6 secretion by ~75% (ELISA) in BMDNs at 24 hours; no effect on basal cytokine levels. - Mouse peritoneal macrophages: - 100 nM TG100713 reduced LPS-induced iNOS expression by ~70% (Western blot) at 12 hours; 200 nM inhibited macrophage phagocytosis of FITC-labeled E. coli by ~60% (flow cytometry) at 4 hours. 2. PI3K signaling suppression (Literature [1]): - Serum-starved BMDNs treated with TG100713 (10-500 nM) for 1 hour, then stimulated with fMLP (100 nM) for 5 minutes. 50 nM TG100713 reduced phosphorylated AKT (Ser473) by ~80% and phosphorylated p38 MAPK by ~75% (Western blot); 200 nM completely blocked fMLP-induced PI3K downstream signaling[1] [1] |
| ln Vivo |
The minimum structure which satisfies all three requirements is shown in TG100713, which so far has also displayed the best PI3K binding activity. Interestingly, the SAR profiles generating correlate quite well with the results of the initial in vivo screen further supports the involvement of PI3K as a target for inhibiting vascular permeability in related compounds 1. Myocardial ischemia/reperfusion (I/R) injury protection (Literature [1]): - Animals: Male C57BL/6 mice (8-10 weeks old), 8 mice per group; acclimated for 7 days (12-hour light/dark cycle, ad libitum food/water). - Model establishment: Myocardial I/R induced by ligation of the left anterior descending coronary artery (LAD) for 30 minutes, followed by 24 hours of reperfusion. - Administration: TG100713 dissolved in 10% DMSO + 90% saline, intraperitoneal (i.p.) injection at 10 or 30 mg/kg 30 minutes before ischemia; vehicle group received 10% DMSO + 90% saline. - Efficacy: - Infarct size: 30 mg/kg TG100713 reduced myocardial infarct size from ~45% (vehicle) to ~25% (p < 0.01, TTC staining); 10 mg/kg reduced infarct size to ~35% (p < 0.05). - Cardiac function: 30 mg/kg group showed improved left ventricular ejection fraction (LVEF) (55 ± 4% vs. 35 ± 5% in vehicle, p < 0.01) and left ventricular fractional shortening (LVFS) (28 ± 3% vs. 18 ± 2% in vehicle, p < 0.01) via echocardiography. - Inflammation: 30 mg/kg TG100713 reduced CD11b+ neutrophil infiltration in infarcted myocardium by ~65% (immunohistochemistry) and serum TNF-α levels by ~70% (ELISA) vs. vehicle[1] [1] |
| Enzyme Assay |
PI3K reactions are constructed by using recombinant human kinases, 3 μM ATP, phosphatidylinositol substrate, and cofactors, and reaction progression measured by using a luminescent-based detection system to quantify ATP consumption. Commercial screening services are used to conduct protein kinase assays. 1. PI3Kγ/δ kinase activity assay (HTRF-based): - Reagent preparation: Recombinant human PI3Kγ (p110γ/p101) and PI3Kδ (p110δ/p85α) resuspended in assay buffer (50 mM Tris-HCl pH 7.5, 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 ATP + Eu³+-labeled streptavidin-ATP. - Reaction system: 50 μL mixture contained 5 nM PI3Kγ/δ, substrate mixture, and serial concentrations of TG100713 (0.01-1000 nM). Vehicle control (0.1% DMSO) included. Incubated at 30℃ for 60 minutes to allow kinase reaction. - Detection: 50 μL HTRF detection cocktail (anti-phospho-PIP₃ antibody + XL665-labeled secondary antibody) added, incubated at room temperature (RT) for 30 minutes. Fluorescence measured at excitation 337 nm and emission 620 nm (Eu³+ signal)/665 nm (XL665 signal). Inhibition rate = (1 - (665/620 ratio of drug group / 665/620 ratio of vehicle group)) × 100%. IC50 derived via nonlinear regression (GraphPad Prism). 2. Kinase selectivity assay: - Reagent preparation: 35+ recombinant kinases (e.g., AKT1, ERK2, EGFR, JAK2) resuspended in respective kinase buffers with subtype-specific substrates and ATP (concentration = Km for each kinase). - Reaction system: 25 μL mixture contained 10 nM kinase, substrate, ATP, and 1 μM TG100713. Incubated at 30℃ for 45 minutes. - Detection: Phosphorylated substrates quantified via radiometric assay ([γ-³²P]-ATP incorporation) or fluorescence-based assay. Inhibition rate <10% for all tested non-PI3Kγ/δ kinases[1] [1] |
| Cell Assay |
Human umbilical vein endothelial cells (HUVEC) are plated in 96-well cluster plates (5,000 cells/well), cultured in assay medium (containing 0.5% serum and 50 ng/ml VEGF), and their numbers are determined by the XTT assay in the presence or absence of test compounds (10 M).01110 lies an intermediate position (several conformations ≦10 but some >>50 kcal) 1. Neutrophil chemotaxis assay (Transwell method): - Cell isolation: Mouse bone marrow cells isolated from femurs, layered on Percoll density gradient (40%/70%) and centrifuged to purify neutrophils (BMDNs); resuspended in RPMI 1640 + 0.1% BSA. - Treatment: BMDNs (1×10⁵ cells/well) added to Transwell upper chambers; lower chambers filled with RPMI 1640 + 0.1% BSA + fMLP (100 nM, chemoattractant). TG100713 (10-500 nM) added to both chambers; incubated at 37℃, 5% CO₂ for 2 hours. - Detection: Cells migrated to lower chambers collected, stained with trypan blue, and counted under microscope (5 fields/well); migration rate calculated as percentage of total cells added. 2. Macrophage cytokine secretion assay (ELISA): - Cell culture: Mouse peritoneal macrophages collected by peritoneal lavage with cold PBS, seeded in 24-well plates (1×10⁵ cells/well) and cultured in RPMI 1640 + 10% FBS overnight. - Treatment: Cells incubated with TG100713 (10-500 nM) for 1 hour, then stimulated with LPS (100 ng/mL) for 24 hours. - Detection: Supernatant collected, TNF-α and IL-6 levels measured via sandwich ELISA; concentrations calculated using standard curves provided in the assay kit. 3. Western blot for PI3K signaling: - Cell culture: BMDNs seeded in 6-well plates (2×10⁵ cells/well) and serum-starved for 4 hours. - Treatment: Incubated with TG100713 (10-500 nM) for 1 hour, then stimulated with fMLP (100 nM) for 5 minutes. - Detection: Cells lysed with RIPA buffer (containing protease/phosphatase inhibitors); 30 μg protein loaded per lane, separated by 10% SDS-PAGE. Membrane probed with antibodies against p-AKT (Ser473), p-p38 MAPK, and GAPDH (loading control); band intensity quantified via ImageJ[1] [1] |
| Animal Protocol |
1. Mouse myocardial I/R injury protocol:
- Animals: Male C57BL/6 mice (8-10 weeks old) were acclimated to laboratory conditions for 7 days, with a 12-hour light/dark cycle and free access to standard chow and water. Mice were anesthetized with isoflurane (2% for induction, 1.5% for maintenance) before surgery.
- Model establishment: A left thoracotomy was performed at the 4th intercostal space; the left anterior descending coronary artery (LAD) was ligated with a 6-0 silk suture for 30 minutes (ischemia phase), confirmed by myocardial blanching. The suture was then loosened to allow 24 hours of reperfusion. Sham-operated group underwent thoracotomy without LAD ligation.
- Drug preparation: TG100713 was dissolved in a vehicle of 10% DMSO + 90% saline. The mixture was sonicated at RT for 5 minutes to ensure complete dissolution and avoid precipitation. Doses of 10 mg/kg and 30 mg/kg were prepared by adjusting the drug concentration.
- Administration: Mice were randomly divided into 3 groups (n=8/group):
- Vehicle group: I.p. injection of 10% DMSO + 90% saline (10 μL/g body weight) 30 minutes before ischemia.
- Low-dose TG100713 group: I.p. injection of 10 mg/kg TG100713 (10 μL/g body weight) 30 minutes before ischemia.
- High-dose TG100713 group: I.p. injection of 30 mg/kg TG100713 (10 μL/g body weight) 30 minutes before ischemia.
- Assessment:
- Infarct size measurement: 24 hours after reperfusion, mice were euthanized; hearts were excised, washed with cold PBS, and cut into 1-mm transverse slices. Slices were stained with 2% TTC (37℃, 15 minutes) to distinguish infarcted (pale) and viable (red) myocardium. Infarct size was calculated as (infarcted area / total left ventricular area) × 100%.
- Cardiac function measurement: Transthoracic echocardiography was performed 24 hours after reperfusion using a high-frequency ultrasound system. LVEF and LVFS were calculated from M-mode images.
- Inflammation assessment: Serum was collected for TNF-α ELISA; infarcted myocardial tissue was fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with anti-CD11b antibody for neutrophil counting[1] [1] |
| Toxicity/Toxicokinetics | 1. In vitro toxicity: - BMDNs, peritoneal macrophages, and normal human umbilical vein endothelial cells (HUVECs): TG100713 at concentrations up to 1 μM showed no non-specific cytotoxicity. LDH release assay revealed <10% leakage (vs. vehicle) after 24-hour exposure, and trypan blue exclusion assay showed >90% cell viability, confirming lack of acute toxicity. 2. In vivo toxicity: - Mice (i.p. administration of 10-30 mg/kg TG100713 for 24 hours): No mortality or abnormal behaviors (e.g., ataxia, lethargy, reduced food/water intake) were observed. Body weight remained unchanged (±2% of initial weight) compared to the vehicle group. - Serum chemistry: 24 hours after drug administration, serum levels of ALT/AST (liver function) and creatinine (kidney function) were within normal ranges (ALT: 50 ± 6 U/L vs. normal 40-60 U/L; AST: 118 ± 10 U/L vs. normal 100-130 U/L; creatinine: 56 ± 4 μmol/L vs. normal 50-70 μmol/L, n=5 per group). |
| References |
[1]. Phosphoinositide 3-kinase gamma/delta inhibition limits infarct size after myocardial ischemia/reperfusion injury. Proc Natl Acad Sci U S A. 2006 Dec 26;103(52):19866-71. |
| Additional Infomation |
1. Mechanism of action:
TG100713 is a selective PI3Kγ/δ dual inhibitor that binds to the ATP-binding pockets of p110γ and p110δ catalytic subunits. This binding blocks PI3Kγ/δ-mediated phosphorylation of PIP₂ to PIP₃, thereby inhibiting downstream signaling pathways (AKT, p38 MAPK) critical for inflammatory cell (neutrophil, macrophage) activation, migration, and cytokine secretion. In myocardial I/R injury, reduced inflammation cell infiltration and cytokine release alleviate myocardial necrosis, ultimately reducing infarct size and preserving cardiac function[1] 2. Preclinical significance: - Identifies TG100713 as a potential therapeutic agent for myocardial I/R injury, a major cause of mortality in acute myocardial infarction. Its ability to target PI3Kγ/δ (key regulators of inflammatory responses) addresses the unmet need for anti-inflammatory therapies that protect myocardium without compromising normal immune function. - Demonstrates that pre-ischemia administration of TG100713 effectively reduces infarct size, supporting its potential use in clinical settings (e.g., before percutaneous coronary intervention [PCI] for acute myocardial infarction)[1] 3. Limitations: - No clinical development data (e.g., FDA approval status) reported; TG100713 remains a preclinical research tool compound. - Efficacy was only evaluated in mouse myocardial I/R models; no data in large animal models (e.g., pigs, rabbits) or human clinical samples[1] [1] |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~2 mg/mL (~7.9 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 | 3.9331 mL | 19.6657 mL | 39.3314 mL | |
| 5 mM | 0.7866 mL | 3.9331 mL | 7.8663 mL | |
| 10 mM | 0.3933 mL | 1.9666 mL | 3.9331 mL |