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Gedatolisib (PF-05212384, PKI-587) is a brand-new, extremely powerful dual inhibitor of PI3Kα, PI3Kγ and mTOR that may have anticancer properties. In cell-free assays, it inhibits PI3Kα, PI3Kγ and mTOR with IC50 values of 0.4 nM, 5.4 nM, and 1.6 nM, respectively. By concentrating on phosphatidylinositol 3 kinase (PI3K) and mammalian target of rapamycin (mTOR) in the PI3K/mTOR signaling pathway, it may have anti-cancer effects. The cellular phosphorylation of PI3K/mTOR signaling pathway proteins is inhibited by PF-05212384. It prevents Akt from being phosphorylated along with the Akt effector proteins GSK3 kinase, ENOS, and PRAS 40. Additionally, in a number of xenograft models, including H1975, BT474, HCT116, H1975, and U87MG, PF-05212384 exhibits strong anti-tumor activity.
Physicochemical Properties
| Molecular Formula | C32H41N9O4 |
| Molecular Weight | 615.7258 |
| Exact Mass | 615.328 |
| Elemental Analysis | C, 62.42; H, 6.71; N, 20.47; O, 10.39 |
| CAS # | 1197160-78-3 |
| Related CAS # | 1197160-78-3 |
| PubChem CID | 44516953 |
| Appearance | white solid powder |
| Density | 1.4±0.1 g/cm3 |
| Index of Refraction | 1.670 |
| LogP | -0.76 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 10 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 45 |
| Complexity | 913 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(C1C([H])=C([H])C(=C([H])C=1[H])N([H])C(N([H])C1C([H])=C([H])C(=C([H])C=1[H])C1=NC(=NC(=N1)N1C([H])([H])C([H])([H])OC([H])([H])C1([H])[H])N1C([H])([H])C([H])([H])OC([H])([H])C1([H])[H])=O)N1C([H])([H])C([H])([H])C([H])(C([H])([H])C1([H])[H])N(C([H])([H])[H])C([H])([H])[H] |
| InChi Key | DWZAEMINVBZMHQ-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C32H41N9O4/c1-38(2)27-11-13-39(14-12-27)29(42)24-5-9-26(10-6-24)34-32(43)33-25-7-3-23(4-8-25)28-35-30(40-15-19-44-20-16-40)37-31(36-28)41-17-21-45-22-18-41/h3-10,27H,11-22H2,1-2H3,(H2,33,34,43) |
| Chemical Name | -[4-[4-(dimethylamino)piperidine-1-carbonyl]phenyl]-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea |
| Synonyms | Gedatolisib; PKI-587; PKI587; PKI 587; PF-05212384; PF05212384; PF 05212384; PF5212384; PF 5212384; PF-5212384 |
| 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 | PI3Kα (IC50 = 0.4 nM); mTOR (IC50 = 1.6 nM); PI3Kγ (IC50 = 5.4 nM) |
| ln Vitro | PKI-587 shows potent inhibitory activity against PI3K-α, PI3K-γ and mTOR with IC50 of 0.4 nM, 5.4 nM and 1.6 nM, respectively. Furthermore, PKI-587 also exhibits its potency against the most frequently occurring mutant forms of PI3Kα, notably the H1047R and E545K with IC50 of 0.6 nM and 0.6 nM, respectively.[1] In MDA-361 and PC3-MM2 cell lines, PKI-587 inhibits tumor cell growth with an IC50 of 4 nM and 13.1 nM, respectively. This effect is correlated with the suppression of phosphorylation of PI3K/mTOR signaling pathway proteins.[1] |
| ln Vivo | PKI-587 treatment at 25 mg/kg intravenously results in low plasma clearance (7 (mL/min)/kg), high volume of distribution (7.2 L/kg), and a lengthy (14.4 hours) half-life in nude mice. With a minimum effective dose (MED) of 3 mg/kg and a maximum tolerated single dose (MTD) of 30 mg/kg against MDA-361 tumors, PKI-587 exhibits strong antitumor efficacy in the MDA-361 xenograft model. While PKI-587 at 25 mg/kg for 7 weeks results in 90% survival of the treated group in the H1975 (non-small-cell lung carcinoma, mutant EGFR [L858R, T790M]) xenograft model. [1] |
| Enzyme Assay | Enzyme assays are done in fluorescent polarization (FP) format, adapted from the Echelon K-1100 PI3K FP assay kit protocol. E545K and H1047R human class I PI3K mutants can be generated in Sf9 or bought from Upstate Biotech. Escherichia coli produces GST-GRP1 (murine), which is then isolated using GST-Sepharose. Assay buffers are reaction buffer [20 mM HEPES (pH 7.1), 2 mM MgCl2, 0.05% CHAPS, and 0.01% β-mercaptoethanol] and stop/detection buffer [100 mM HEPES (pH 7.5), 4 mM EDTA, 0.05% CHAPS]. FP reaction is run for 30 minutes at room temperature in 20 μL of reaction buffer containing 20 μM phosphatidylinositol 4,5-bisphosphate (PIP2), 25 μM ATP, and<4% DMSO. FP reaction is stopped with 20 μL of stop/detection buffer (10 nM probe and 40 nM GST-GRP), and after 2 hours, data are collected using an Envision plate reader. The routine assays with purified FLAG-TOR (FL and 3.5) are performed in 96-well plates as follows. Enzymes are first diluted in kinase assay buffer (10 mM Hepes (pH 7.4), 50 mM NaCl, 50 mM β-glycerophosphate, 10 mM MnCl2, 0.5 mM DTT, 0.25 μM microcystin LR, and 100 μg/mL BSA). To each well, 12 μL of the diluted enzyme is mixed briefly with 0.5 μL test inhibitor or control vehicle dimethyl sulfoxide (DMSO). The kinase reaction is initiated by adding 12.5 μL kinase assay buffer containing ATP and His6-S6K to give a final reaction volume of 25 μL containing 800 ng/mL FLAG-TOR, 100 μM ATP, and 1.25 μM His6-S6K. The reaction plate is incubated for 2 hours (linear at 1–6 hours) at room temperature with gentle shaking and then terminated by adding 25 μL Stop buffer (20 mM Hepes (pH 7.4), 20 mM EDTA, and 20 mM EGTA). |
| Cell Assay | Cells are plated in 96-well culture plates at about 3000 cells per well. PKI-587 is added to cells a day after plating. Viable cell densities are assessed three days after PKI-587 treatment by monitoring the metabolic conversion of the dye MTS (by viable cells), a tried-and-true cell proliferation assay. Freshly thawed and mixed MTS and PMS stocks (20:1) are used for each assay. Following this, 20 L of the MTS/PMS mixture is added to 96-well cell plates, and the plates are incubated for 1 to 2 hours in a cell culture incubator. |
| Animal Protocol |
MDA-361 and H1975 cells are injected subcutaneously into the nude mice. ≤30 mg/kg Administered via i.v. |
| References |
[1]. J Med Chem . 2010 Mar 25;53(6):2636-45. [2]. J Surg Res . 2012 Aug;176(2):542-8. |
| Additional Infomation |
Gedatolisib has been used in trials studying the basic science and treatment of Neoplasm, Ovary Cancer, Breast Cancer, Advanced Cancer, and Endometrial Cancer, among others. Gedatolisib is an agent targeting the phosphatidylinositol 3 kinase (PI3K) and mammalian target of rapamycin (mTOR) in the PI3K/mTOR signaling pathway, with potential antineoplastic activity. Upon intravenous administration, gedatolisib inhibits both PI3K and mTOR kinases, which may result in apoptosis and growth inhibition of cancer cells overexpressing PI3K/mTOR. Activation of the PI3K/mTOR pathway promotes cell growth, survival, and resistance to chemotherapy and radiotherapy; mTOR, a serine/threonine kinase downstream of PI3K, may also be activated independent of PI3K. |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~2 mg/mL (~3.2 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 | 1.6241 mL | 8.1204 mL | 16.2409 mL | |
| 5 mM | 0.3248 mL | 1.6241 mL | 3.2482 mL | |
| 10 mM | 0.1624 mL | 0.8120 mL | 1.6241 mL |