| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Akt1 (IC₅₀ = 5 nM); Akt2 (IC₅₀ = 18 nM); Akt3 (IC₅₀ = 8 nM); Highly selective against 339 kinases (only ROCK2, PRKGB, and Aurora A showed >50% inhibition at 1 μM)
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|---|---|
| ln Vitro |
Ipatasertib tosylate (10 µM; 12, 24 h) inhibits colon cancer cell proliferation through p53-independent PUMA activation in cell-based experiments[1]. Ipatasertib tosylate (1, 5, 10, 20 μM; 24 h/10 μM; 3, 6, 12, 24 h) upregulates PUMA expression in HCT116 cells in a time- and concentration-dependent manner[1]. Ipatasertib tosylate increases PUMA mRNA levels in wild-type, p53−/−, and DLD1 (p53 mutant) HCT116 cells[1]. Ipatasertib tosylate (10 µM; 24 h) induces apoptosis of HCT116 cells via the PUMA/Bax pathway[1].
Ipatasertib dose-dependently inhibits colon cancer cell proliferation (HCT116 IC₅₀ ≈10 μM) and induces apoptosis via PUMA upregulation, independent of p53 status. It activates FoxO3a and NF-κB, both binding to the PUMA promoter to trigger Bax-mediated mitochondrial apoptosis. FoxO3a is the primary regulator (ChIP assay shows stronger binding), while NF-κB is secondary. Synergistic effects observed with 5-FU, cisplatin, and regorafenib, further enhancing PUMA-dependent apoptosis. |
| ln Vivo |
Ipatasertib tosylate (30 mg/kg; po; single daily for 15 consecutive days) showed PUMA-dependent antitumor activity in wild-type and PUMA−/− HCT116 xenograft mouse models [1].
Ipatasertib (30 mg/kg/day oral gavage) significantly suppresses HCT116 WT xenograft tumor growth in mice (tumor volume/weight reduction), but efficacy is diminished in PUMA-KO tumors. Immunohistochemistry confirms reduced Ki67 (proliferation marker) and increased cleaved caspase-3 in WT tumors only. |
| Enzyme Assay |
Akt inhibition measured by TR-FRET assay using recombinant Akt enzymes and biotinylated Crosstide substrate.
Western blotting detects Akt inhibition (↓p-Akt S473), FoxO3a activation (↓p-FoxO3a S253), NF-κB activation (↑p-p65 S536), and PUMA/Bax expression. ChIP assay confirms FoxO3a/NF-κB binding to PUMA promoter. |
| Cell Assay |
Cell Viability Assay[1]
Cell Types: HCT116 WT, p53−/−, and DLD1 (p53 mutant) Tested Concentrations: 10 µM Incubation Duration: 12, 24 h Experimental Results: Decreased all the three cell lines viability. Apoptosis Analysis[1] Cell Types: HCT116 Tested Concentrations: 10 µM Incubation Duration: 24 h Experimental Results: Induced apoptosis through PUMA/Bax pathway. Western Blot Analysis[1] Cell Types: HCT116 WT, p53−/−, and DLD1 (p53 mutant) Tested Concentrations: 1, 5, 10, 20 μM for 24 h/10 μM for 3, 6, 12, 24 h Incubation Duration: 24 h; 3, 6, 12, 24 h Experimental Results: Increased the level of PUMA in a concentration and time dependent manner. Proliferation: CCK-8 assay after 24–72h treatment; IC₅₀ calculated in HCT116, DLD1, and p53-KO cells. Apoptosis: Hoechst 33258 staining for nuclear condensation; flow cytometry (Annexin V/PI) for apoptosis quantification. PUMA dependency: PUMA/Bax-KO cells abolish apoptosis; colony formation assays confirm growth inhibition requires PUMA/Bax. |
| Animal Protocol |
Animal/Disease Models: HCT116 WT and PUMA−/− cells xenograft nude mice model[1].
Doses: 30 mg/kg Route of Administration: Oral gavage; single daily for 15 consecutive days. Experimental Results: Inhibited growth of tumors in a PUMA-dependent manner. Xenograft: Nude mice implanted with HCT116 WT or PUMA-KO cells; treated with ipatasertib (30 mg/kg/day oral gavage) for 15 days. Endpoints: Tumor volume/weight measurement; IHC for P-Akt, Ki67, cleaved caspase-3. |
| ADME/Pharmacokinetics |
Bioavailability: Mice 31%, Rats 80%, Dogs 44%, Humans ~60%
Half-life: Mice 2.3 hours, Rats 3.1 hours, Dogs 7.8 hours, Humans ~50 hours Metabolism: Primarily via CYP3A4-mediated reduction of ketone bodies to alcohol metabolites (inactive); renal excretion <5%. Plasma protein binding: >95% for all species; Volume of distribution (Vd) = 1.1-1.7 L/kg. |
| Toxicity/Toxicokinetics |
At doses ≥100 mg/kg/day, reversible increases in ALT/AST were observed in rats/dogs.
No QTc interval prolongation was observed in canine telemetry (30 mg/kg). Maximum tolerated clinical dose: 600 mg/day; dose-limiting toxicities include diarrhea and hyperglycemia. Drug interactions: Potent CYP3A4 inhibitors increased AUC by 5.7-fold; inducers reduced exposure by 12-fold. At therapeutic doses, toxicity to normal colon cells (NCM460) is extremely low. |
| References | |
| Additional Infomation |
Mechanism: An ATP-competitive pan-Akt inhibitor that blocks downstream signaling pathways. Clinical Indication: Phase II/III clinical trials for triple-negative breast/prostate cancer (e.g., IPATUnity130). Tosylate was selected for optimal solubility (free base 0.5 mg/mL, tosylate 8.7 mg/mL). Key Differentiating Factor: In vivo target binding studies can be conducted using pPRAS40 inhibition as a pharmacodynamic biomarker. Colorectal cancer is one of the three most common malignancies with low survival rates. Ipatasertib is a novel, highly selective ATP-competitive pan-Akt inhibitor that has shown potent antitumor activity in various cancers, including colorectal cancer. However, the exact underlying mechanism for its clinical treatment of colorectal cancer remains unclear. This study aims to determine whether ipatasertib inhibits colorectal cancer growth through PUMA-dependent apoptosis. Ipatasenet inhibits Akt, leading to p53-independent PUMA activation, which in turn simultaneously activates FoxO3a and NF-κB. The latter directly binds to the PUMA promoter, upregulating PUMA transcription and inducing Bax-mediated mitochondrial endogenous apoptosis. Notably, the Akt/FoxO3a/PUMA pathway is the primary pathway for ipatazepet-induced PUMA activation in colorectal cancer, while the Akt/NF-κB/PUMA pathway is secondary. PUMA knockout eliminates ipatazepet-induced apoptosis both in vitro and in vivo (xenograft tumors). Furthermore, PUMA is indispensable in the combination therapy of ipatazepet with certain traditional or novel drugs. In conclusion, our study demonstrates that FoxO3a and NF-κB-induced PUMA activation is a key step in the inhibition of colorectal cancer growth by ipatazepet treatment, providing a theoretical basis for clinical evaluation.
|
| Molecular Formula |
C31H40CLN5O5S
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|---|---|
| Molecular Weight |
630.20
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| CAS # |
1491138-24-9
|
| Appearance |
Typically exists as solids at room temperature
|
| SMILES |
C[C@@H]1C[C@H](C2=C1C(=NC=N2)N3CCN(CC3)C(=O)[C@H](CNC(C)C)C4=CC=C(C=C4)Cl)O.CC1=CC=C(C=C1)S(=O)(=O)O
|
| Synonyms |
GDC-0068 tosylate; Ipatasertib (tosylate); SCHEMBL16617584; UTUKVTLAXHISRJ-GJYOXNSLSA-N; (S)-2-(4-chlorophenyl)-1-(4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperazin-1-yl)-3-(isopropylamino)propan-1-one p-toluene Sulfonic Acid Salt; 1491138-24-9; RG7440 tosylate
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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 | 1.5868 mL | 7.9340 mL | 15.8680 mL | |
| 5 mM | 0.3174 mL | 1.5868 mL | 3.1736 mL | |
| 10 mM | 0.1587 mL | 0.7934 mL | 1.5868 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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