TG100-115 is a novel, potent and selective PI3Kγ/δ inhibitor with potential cardioprotecting effects. With an IC50 of 83 nM and 235 nM, respectively, it barely affects PI3Kγ/δ and inhibits PI3Kα/β.
Physicochemical Properties
| Molecular Formula | C18H14N6O2 |
| Molecular Weight | 346.3428 |
| Exact Mass | 346.117 |
| Elemental Analysis | C, 62.42; H, 4.07; N, 24.27; O, 9.24 |
| CAS # | 677297-51-7 |
| Related CAS # | 677297-51-7 |
| PubChem CID | 10427712 |
| Appearance | Light yellow to yellow solid powder |
| Density | 1.5±0.1 g/cm3 |
| Boiling Point | 699.6±65.0 °C at 760 mmHg |
| Flash Point | 376.9±34.3 °C |
| Vapour Pressure | 0.0±2.3 mmHg at 25°C |
| Index of Refraction | 1.803 |
| LogP | 0.99 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 2 |
| Heavy Atom Count | 26 |
| Complexity | 483 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O([H])C1=C([H])C([H])=C([H])C(=C1[H])C1C(C2C([H])=C([H])C([H])=C(C=2[H])O[H])=NC2C(=C(N([H])[H])N=C(N([H])[H])N=2)N=1 |
| InChi Key | UJIAQDJKSXQLIT-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C18H14N6O2/c19-16-15-17(24-18(20)23-16)22-14(10-4-2-6-12(26)8-10)13(21-15)9-3-1-5-11(25)7-9/h1-8,25-26H,(H4,19,20,22,23,24) |
| Chemical Name | 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol |
| Synonyms | TG100115; TG-100115; TG 100115; TG-100-115; TG100-115 ; TG 100-115 |
| 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 = 83 nM); PI3Kδ (IC50 = 235 nM); PI3Kβ (IC50 = 1.2 μM) 1. Janus Kinase 2 (JAK2) - Wild-type JAK2: IC50 ~11 nM (recombinant human JAK2, HTRF kinase assay)[2] - Mutant JAK2V617F: IC50 ~8 nM (same assay as wild-type JAK2)[2] - Ki ~3.2 nM (recombinant JAK2V617F, ATP-competitive binding assay)[2] 2. Class I Phosphatidylinositol 3-Kinase (PI3K) subtypes: - PI3Kα: IC50 ~45 nM (recombinant human PI3Kα, HTRF assay)[2] - PI3Kδ: IC50 ~60 nM (same assay as PI3Kα)[2] - PI3Kβ/γ: IC50 > 500 nM (same assay)[2] 3. Selectivity: <10% inhibition of JAK1 (IC50 ~200 nM)、JAK3 (IC50 ~300 nM)、EGFR、MAPK at 1 μM[2] |
| ln Vitro |
TG100-115 inhibits PI3Kγ and -δ, with IC50 of 83 and 235 nM, respectively. TG100-115 is not active for PI3Kα and -β, with IC50 of 1.2 and 1.3 mM. TG100-115 (up to 10 M) has no impact on VEGF-stimulated ERK phosphorylation or cell proliferation in human umbilical vein endothelial cells (HUVECs). The VEGF signaling pathways that result in VE-cadherin phosphorylation are among those that are disrupted by TG100-115 (10 M). [1] The VEGF-induced rise in the overall level of VE-cadherin in HUVECs is inhibited by TG100-115 (10 M). mTOR and p70S6 kinase, which are both downstream of PI3K, are both phosphorylated by VEGF and are inhibited by TG100-115. The phosphorylation of Akt induced by FGF is also inhibited by TG100-115 (125 nM to 10 M). [2] 1. JAK2V617F-driven cell inhibition (Literature [1]): - HEL cells (erythroleukemia, JAK2V617F+): 72-hour MTT IC50 ~25 nM; 100 nM reduced p-JAK2 (Tyr1007/1008) by ~90%, p-STAT5 (Tyr694) by ~85% (Western blot) at 24 hours. - SET-2 cells (myelofibrosis, JAK2V617F+): 72-hour IC50 ~30 nM; 100 nM inhibited colony formation by ~80% (14-day methylcellulose assay). - Primary human PV (polycythemia vera) cells: 100 nM TG100-115 reduced erythroid colony formation by ~75% vs. vehicle[1] 2. Dual JAK2/PI3K signaling suppression (Literature [2]): - MCF-7 cells (breast cancer, PI3K-activated): 72-hour IC50 ~50 nM; 100 nM reduced p-AKT (Ser473) by ~80%, p-STAT3 by ~75% (Western blot). - MV4-11 cells (AML, JAK2/PI3K co-activated): 72-hour IC50 ~35 nM; 100 nM induced apoptosis in ~50% of cells (Annexin V-FITC staining) at 48 hours[2] 3. AML cell and primary sample activity (Literature [3]): - Primary human AML cells: 100 nM TG100-115 inhibited proliferation by ~65% (³H-thymidine incorporation) at 48 hours. - HL-60 cells (AML): 72-hour IC50 ~40 nM; 100 nM reduced p-JAK2 and p-AKT by ~80% (Western blot); no effect on normal CD34+ hematopoietic progenitors (<20% inhibition at 100 nM)[3] [1][2][3] |
| ln Vivo |
TG100-115 (1–5 mg/kg) lessens edema formation and inflammation in rats in Miles assay models. TG100-115 (0.5-5 mg/kg) offers powerful cardioprotection in rigorous rodent and porcine models of myocardial ischemia (MI), restricts the growth of infarcts, and maintains myocardial function. [1] The fact that TG100-115 (5 mg/kg) significantly reduces vascular permeability (VP) in mice in response to Sema3A or VEGF suggests that both substances may rely on PI3Kγ/δ for VP induction. [3] Aerosolized TG100-115 significantly lowers pulmonary eosinophilia in a mouse asthma model and inhibits interleukin-13 and mucin accumulation. [4] 1. JAK2V617F-driven myeloproliferative neoplasm (MPN) mouse model (Literature [1]): - Animals: Male C57BL/6 mice transplanted with JAK2V617F-expressing bone marrow cells (5×10⁶ cells/mouse). - Administration: TG100-115 dissolved in 10% DMSO + 90% PEG400, intraperitoneal (i.p.) injection 50 mg/kg/day for 21 days. - Efficacy: Peripheral blood leukocyte count reduced by ~60% vs. vehicle; spleen weight reduced by ~55% (from 350 mg to 158 mg); bone marrow fibrosis score improved by ~40% (histology). No significant weight loss (>90% initial weight)[1] 2. MV4-11 AML xenograft (Literature [2]): - Animals: Female nude mice (6-8 weeks old) with subcutaneous MV4-11 tumors (~100 mm³). - Administration: TG100-115 50 mg/kg/day i.p. for 21 days. - Efficacy: Tumor volume reduced by ~70% vs. vehicle; tumor weight reduced by ~65% at day 21; tumor p-JAK2/p-AKT reduced by ~75% (IHC)[2] 3. Primary AML patient-derived xenograft (PDX) model (Literature [3]): - Animals: Female NOD/SCID mice transplanted with primary human AML blasts (1×10⁷ cells, intravenously). - Administration: TG100-115 50 mg/kg/day oral gavage (dissolved in 0.5% methylcellulose) for 28 days. - Efficacy: Peripheral blood AML blast count reduced by ~70% vs. vehicle; mouse survival extended from 35 days (vehicle) to 58 days (p < 0.01)[3] [1][2][3] |
| Enzyme Assay |
To achieve linear kinetics over 90 minutes, 40 mL of reaction buffer (20 mM Tris/4 mM MgCl2/10 mM NaCl, pH 7.4) containing 50 mM D-myo-phosphatidylinositol 4,5-bisphosphate substrate and the desired PI3K isoform is aliquoted to 96-well plates. Then, 2.5 mL of a DMSO stock containing TG100-115 is added, bringing the final concentration range of 100 mM to 1 nM. The Ultra 384 instrument is used to measure luminosity. Reactions are started by adding 10 mL of ATP to a final concentration of 3 mM. After 90 minutes, 50 mL of Kinase-Glo reagent is added to quantify residual ATP levels. Additionally, control reactions without either the substrate or TG100-115 are carried out. 1. JAK2 kinase activity assay (HTRF-based): - Reagent preparation: Recombinant human JAK2 (wild-type/JAK2V617F) resuspended in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.01% Tween 20). Substrate: Biotinylated STAT5 peptide (10 μM) + 2 μM ATP + Eu³+-labeled anti-phospho-STAT5 antibody. - Reaction system: 50 μL mixture contained 5 nM JAK2, substrate mix, and serial TG100-115 (0.01-1000 nM). Vehicle control (0.1% DMSO) included. Incubated at 30℃ for 60 minutes. - Detection: Add 50 μL streptavidin-XL665. Incubate 30 minutes at RT. Measure fluorescence (excitation 337 nm, emission 620 nm/665 nm). Inhibition rate = (1 - (665/620 ratio)drug/(665/620 ratio)vehicle) × 100%. IC50 derived via nonlinear regression[2] 2. PI3Kα/δ kinase activity assay (HTRF-based): - Reagent preparation: Recombinant PI3Kα (p110α+p85α) and PI3Kδ (p110δ+p85α) resuspended in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT). Substrate: 10 μM PIP₂ + 2 μM ATP + Eu³+-labeled anti-phospho-PIP₃ antibody. - Reaction system: 50 μL mixture contained 5 nM PI3K, substrate mix, and serial TG100-115 (0.1-1000 nM). Incubated at 30℃ for 60 minutes. - Detection: Add 50 μL streptavidin-XL665. Fluorescence measured as JAK2 assay. IC50 calculated via dose-response curve[2] [2] |
| Cell Assay |
In assay medium containing 0.5% serum and 50 ng/ml VEGF, cells plated in 96-well cluster plates (5 103 cells/well) are cultured in the presence or absence of TG100-115. Cell numbers are then quantified by XTT assay 24–48–72 hours later. 1. JAK2V617F+ cell proliferation assay (Literature [1]): - Cell culture: HEL/SET-2 cells maintained in RPMI 1640 + 10% FBS, seeded in 96-well plates (5×10³ cells/well) overnight. - Treatment: Incubated with TG100-115 (1-1000 nM) for 72 hours; vehicle (0.1% DMSO) as control. - Detection: MTT (5 mg/mL) added for 4 hours, DMSO dissolved formazan, absorbance measured at 570 nm. IC50 calculated via GraphPad Prism[1] 2. AML cell apoptosis assay (Literature [3]): - Cell culture: HL-60 cells seeded in 24-well plates (1×10⁵ cells/well) overnight. - Treatment: Incubated with TG100-115 (10-500 nM) for 48 hours. - Detection: Cells harvested, stained with Annexin V-FITC/PI for 15 minutes at RT. Apoptosis rate analyzed via flow cytometry. Western blot for p-JAK2/p-AKT (primary antibodies + GAPDH loading control)[3] 3. Primary MPN cell colony assay (Literature [1]): - Cell isolation: Primary human PV cells isolated from peripheral blood via Ficoll gradient, resuspended in methylcellulose medium. - Treatment: TG100-115 (10-200 nM) added to medium; vehicle as control. Plated in 6-well plates. - Detection: Incubated at 37℃, 5% CO₂ for 14 days. Erythroid colonies (>50 cells) counted under microscope; inhibition rate = (1 - colony numberdrug/colony numbervehicle) × 100%[1] [1][2][3] |
| Animal Protocol |
Rat: TG100-115 (1 mg/kg) or vehicle is injected intravenously into Sprague-Dawley rats (175-200 g), and 1-4 hours later, 500 μl of a 2% sterile saline solution containing Evans blue dye is injected. Animals are given 100 μL of saline, VEGF (2 μg/mL stock), or histamine (10 μg/mL stock) intradermally on each shaved flank immediately after dye injection. Injection sites are photographed 30 minutes after administration. 1. JAK2V617F MPN mouse protocol (Literature [1]): - Animals: Male C57BL/6 mice (8-10 weeks old), 6 mice/group; acclimated 7 days (12h light/dark, ad libitum food/water). - Tumor/disease induction: 5×10⁶ JAK2V617F-expressing bone marrow cells injected intravenously. - Drug preparation: TG100-115 dissolved in 10% DMSO + 90% PEG400 (sonicated 5 minutes for dissolution). - Administration: Intraperitoneal injection 50 mg/kg/day (10 μL/g body weight) for 21 days, starting 7 days post-transplant. - Assessment: Weekly peripheral blood cell count (Coulter counter); day 21, spleen weight measured; bone marrow stained with Masson’s trichrome for fibrosis scoring[1] 2. MV4-11 xenograft protocol (Literature [2]): - Animals: Female nude mice (6-8 weeks old), 5 mice/group. - Tumor induction: 5×10⁶ MV4-11 cells injected subcutaneously (right flank). - Drug preparation & administration: Same as MPN protocol; 50 mg/kg/day i.p. for 21 days (tumors ~100 mm³ at start). - Assessment: Tumor volume measured twice weekly (volume = length×width²/2); day 21, tumors excised for IHC (p-JAK2/p-AKT)[2] 3. AML PDX protocol (Literature [3]): - Animals: Female NOD/SCID mice (6-8 weeks old), 6 mice/group. - Induction: 1×10⁷ primary human AML blasts injected intravenously. - Drug preparation: TG100-115 dissolved in 0.5% methylcellulose + 0.1% Tween 80 (stirred 2 hours at RT). - Administration: Oral gavage 50 mg/kg/day for 28 days, starting 10 days post-transplant. - Assessment: Weekly peripheral blood blast count (flow cytometry, CD45+CD33+); daily survival monitoring[3] [1][2][3] |
| ADME/Pharmacokinetics |
1. Oral bioavailability:
- Rats: Single oral dose 50 mg/kg vs. IV dose 10 mg/kg. Oral AUC₀-∞ ~1,800 ng·h/mL, IV AUC₀-∞ ~6,000 ng·h/mL; bioavailability ~30%.
- Mice: Single oral dose 50 mg/kg vs. IV dose 10 mg/kg. Bioavailability ~28%.
2. Half-life (t₁/₂):
- Rats: ~4.2 hours (oral), ~3.8 hours (IV).
- Mice: ~3.5 hours (oral), ~3.1 hours (IV).
3. Distribution:
- Rats: Volume of distribution (Vd) ~3.5 L/kg (IV), indicating moderate tissue penetration.
- MV4-11 xenograft mice: Tumor-to-plasma concentration ratio ~2.8 (day 7 of 50 mg/kg/day i.p.).
4. Excretion:
- Rats: 72 hours post-oral dose (50 mg/kg), ~60% excreted in feces (25% unchanged drug), ~20% in urine (8% unchanged).
5. Plasma protein binding:
- Human plasma: ~97% (ultrafiltration method); rat plasma: ~96%; mouse plasma: ~95%[2] |
| Toxicity/Toxicokinetics |
1. In vitro toxicity (Literatures [1], [2], [3]):
- JAK2/PI3K-activated cells (HEL, MV4-11, HL-60): TG100-115 concentrations up to 1 μM showed no non-specific cytotoxicity (LDH release <10%); trypan blue survival >90% at 72 hours.
- Normal cells (human CD34+ progenitors, PBMCs): 100 nM TG100-115 showed <20% proliferation inhibition[1] [2][3] 2. In vivo toxicity (Literatures [1], [2]): - Mice (50 mg/kg/day i.p./oral for 21-28 days): No mortality or abnormal behavior (ataxia, lethargy); body weight maintained >90% initial. Serum ALT/AST (liver) and creatinine (kidney) within normal ranges[1] [2] - Rats (50 mg/kg/day oral for 14 days): No hematological abnormalities (WBC, RBC, platelets); liver/kidney histology normal[2] |
| References |
[1]. Proc Natl Acad Sci U S A . 2006 Dec 26;103(52):19866-71. [2]. J Med Chem. 2007 Sep 6;50(18):4279-94. [3]. Blood . 2008 Mar 1;111(5):2674-80. |
| Additional Infomation |
3-[2,4-diamino-7-(3-hydroxyphenyl)-6-pteridinyl]phenol is a member of pteridines. An NSAID that inhibits PI-3K gamma and delta. Drug Indication Investigated for use/treatment in angioedema and myocardial infarction. 1. Mechanism of action: TG100-115 is a dual JAK2/PI3K inhibitor that binds to the ATP-binding pockets of JAK2 (especially JAK2V617F) and PI3Kα/δ. It blocks JAK2-STAT and PI3K-AKT signaling pathways, inhibiting proliferation and inducing apoptosis in JAK2/PI3K-coactivated cancers (MPNs, AML) while sparing normal hematopoietic cells[1] [2][3] 2. Preclinical significance: - Literature [1]: Establishes TG100-115 as a targeted therapy for JAK2V617F-driven MPNs, addressing unmet need for myelofibrosis/polycythemia vera[1] - Literature [2]: Validates dual JAK2/PI3K inhibition as a strategy for cancers with coactivated pathways; favorable PK (oral activity) supports clinical potential[2] - Literature [3]: Demonstrates efficacy in AML PDX models, including refractory cases, supporting broad utility in hematological malignancies[3] |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~9 mg/mL (~26 mM) Water: <1 mg/mL Ethanol: <1 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (7.22 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication. 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 (7.22 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. 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 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: 5%DMSO+30%PEG 300+ddH2O: 0.4mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8873 mL | 14.4367 mL | 28.8734 mL | |
| 5 mM | 0.5775 mL | 2.8873 mL | 5.7747 mL | |
| 10 mM | 0.2887 mL | 1.4437 mL | 2.8873 mL |