| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 50mg | |||
| Other Sizes |
| Targets |
Allosteric inhibitor of inactive AKT kinase (also known as protein kinase B or PKB). It binds to and stabilizes the inactive form of the enzyme, preventing its activation. [1]
|
|---|---|
| ln Vitro |
AKT-IN-1 has an IC50 of 0.422 and 0.322 μM, respectively, which allows it to effectively block the phosphorylation of AKT Thr308 and Ser473 sites in cells. Because ribosomal protein S6 is a downstream effector of the PI3K-AKT pathway, AKT-IN-1 prevents it from becoming phosphorylated. PRAS40 phosphorylation is efficiently inhibited by AKT-IN-1 [1].
Compound 26 demonstrated potent inhibition of AKT enzyme activity with an IC50 of 1.04 μM. [1] In BT474c breast adenocarcinoma cells, compound 26 potently inhibited the phosphorylation of AKT at both its regulatory sites, Thr308 (IC50 = 0.422 μM) and Ser473 (IC50 = 0.322 μM). This is consistent with its mechanism as a prevention of activation inhibitor. [1] It also inhibited the phosphorylation of PRAS40, a downstream substrate of AKT, in a concentration-dependent manner. [1] Furthermore, compound 26 inhibited the phosphorylation of ribosomal protein S6, a downstream effector of the PI3K-AKT pathway, confirming its pathway modulation in cells. [1] It exhibited a logD of 1.3 and showed exceptional free drug levels (41.9% free in rat plasma protein binding assay). [1] Its aqueous solubility at pH 7.4 was measured at 270 μM. [1] |
| ln Vivo |
Compound 26 (AKT-IN-1) in vivo effects were evaluated by assessing the compound's pharmacodynamic efficacy in the BT474c breast cancer xenograft model. AKT-IN-1 efficiently prevented the phosphorylation of AKT (Ser473) and its downstream substrate GSK3β after acute dosages of 100 and 200 mg/kg. Its effectiveness was in line with its pharmacokinetic properties. By assessing the impact on the growth of tumor cell xenografts, the in vivo activity of AKT-IN-1 was further described. AKT-IN-1 (100 and 200 mg/kg) given orally, continuously, and daily to nude mice containing BT474c mammary adenocarcinoma xenografts suppresses tumor growth in a dose-dependent manner. At a daily dosage of 200 mg/kg, AKT-IN-1 significantly inhibits the growth of tumors [1].
In a BT474c breast adenocarcinoma xenograft model in nude mice, oral dosing of compound 26 resulted in inhibition of tumor growth in a dose-dependent manner. At a dose of 200 mg/kg daily, it caused significant tumor growth inhibition. [1] Following acute oral doses of 100 and 200 mg/kg, compound 26 potently inhibited the phosphorylation of its downstream substrate GSK3β, as well as the phosphorylation of AKT at Ser473, in a manner consistent with its pharmacokinetic profile. [1] |
| Enzyme Assay |
The enzymatic activity of AKT1 was assayed for all analogues synthesized. The assay utilized active enzyme and was designed specifically for testing inhibitors of catalysis. However, the text notes that this assay can detect activity via both modes of inhibition (ATP-competitive and allosteric), and the quoted activity (IC50 = 1.04 μM for compound 26) is therefore an underestimate of the true potency for an allosteric inhibitor like compound 26. Cell activity was generally used to monitor chemistry progress. [1]
|
| Cell Assay |
Cell Assay for p-AKT and p-PRAS40: The biological activity and pharmacology of compound 26 were characterized in the BT474c breast adenocarcinoma cell line. Cells were treated with varying concentrations of compound 26 for 24 hours. The phosphorylation status of AKT pathway proteins, including AKT (Ser473 and Thr308), its downstream substrate PRAS40, and ribosomal protein S6, was measured, likely by Western blot or similar immunoassay. Compound 26 potently inhibited the phosphorylation of all these proteins. [1]
Cell Assay for p-AKT T308 and S473 IC50: The IC50 values for inhibition of phosphorylation of Thr308 and Ser473 on AKT were determined in specific cell lines. p-AKT T308 IC50 was measured in BT474 cells and found to be 0.422 μM. p-AKT S473 IC50 was measured in MDAMB468 cells and found to be 0.322 μM. [1] |
| Animal Protocol |
Pharmacodynamic (PD) Study:** The in vivo pharmacodynamic activity of compound 26 was characterized in a BT474c breast adenocarcinoma xenograft model. Tumor-bearing nude mice received acute oral doses of compound 26 at 100 and 200 mg/kg. Following dosing, tumors were harvested and analyzed for the inhibition of phosphorylation of downstream substrates (GSK3β) and AKT itself (Ser473). The effects were correlated with the compound's pharmacokinetic profile. [1]
* **Efficacy (Tumor Growth Inhibition) Study:** The anti-tumor efficacy of compound 26 was evaluated in nude mice bearing BT474c breast adenocarcinoma xenografts. Animals were dosed orally on a continuous (daily) schedule with compound 26 at 100 and 200 mg/kg. Tumor growth was measured over time to assess inhibition. At 200 mg/kg daily, compound 26 caused significant tumor growth inhibition. [1] Pharmacodynamic (PD) Study: The in vivo pharmacodynamic activity of compound 26 was characterized in a BT474c breast adenocarcinoma xenograft model. Tumor-bearing nude mice received acute oral doses of compound 26 at 100 and 200 mg/kg. Following dosing, tumors were harvested and analyzed for the inhibition of phosphorylation of downstream substrates (GSK3β) and AKT itself (Ser473). The effects were correlated with the compound's pharmacokinetic profile. [1] Efficacy (Tumor Growth Inhibition) Study: The anti-tumor efficacy of compound 26 was evaluated in nude mice bearing BT474c breast adenocarcinoma xenografts. Animals were dosed orally on a continuous (daily) schedule with compound 26 at 100 and 200 mg/kg. Tumor growth was measured over time to assess inhibition. At 200 mg/kg daily, compound 26 caused significant tumor growth inhibition. [1] |
| ADME/Pharmacokinetics |
Compound 26 showed favorable DMPK properties. [1]
It exhibited very low intrinsic clearance in both rat and human in vitro hepatocyte assays (Clint = 1 μL/min/10⁶ cells for rat hepatocytes; data for human hepatocytes is not individually listed in Table 4 but the text describes it as having the lowest clearance in both assays). [1] In rat in vivo studies, it demonstrated low blood clearance (Clb = 32 mL/min/kg), a moderate volume of distribution (Vdss = 1.1 L/kg), and good oral bioavailability of 48%. [1] It has a logD of 1.3 and a high free fraction in rat plasma (41.9% free). [1] |
| References | |
| Additional Infomation |
Compound 26 is a pyridine amide derivative synthesized as part of a medicinal chemistry campaign to discover novel allosteric AKT inhibitors with improved potency and physicochemical properties. [1]
Its design was based on a strategy to explore alternative core scaffolds to the known quinoline-based allosteric AKT inhibitors. [1] The combination of good potency, very high free drug levels, and excellent oral bioavailability made it a candidate for further in vivo efficacy profiling. [1] It was shown to be an allosteric inhibitor that prevents the activation of AKT, leading to decreased phosphorylation of AKT and its downstream targets (PRAS40, GSK3β, S6). [1] The study concludes that compound 26 represents a successful example of a novel scaffold with a good balance of in vitro potency, physicochemical properties, and in vivo efficacy. [1] |
| Molecular Formula |
C22H21N3O
|
|---|---|
| Molecular Weight |
343.43
|
| Exact Mass |
343.168
|
| CAS # |
1357158-81-6
|
| PubChem CID |
56953649
|
| Appearance |
White to off-white solid powder
|
| Density |
1.2±0.1 g/cm3
|
| Boiling Point |
496.4±45.0 °C at 760 mmHg
|
| Flash Point |
254.0±28.7 °C
|
| Vapour Pressure |
0.0±1.3 mmHg at 25°C
|
| Index of Refraction |
1.643
|
| LogP |
2.87
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
26
|
| Complexity |
490
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
GIRZDHCBMNHMEH-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C22H21N3O/c23-21(26)17-13-19(15-5-2-1-3-6-15)20(25-14-17)16-7-9-18(10-8-16)22(24)11-4-12-22/h1-3,5-10,13-14H,4,11-12,24H2,(H2,23,26)
|
| Chemical Name |
6-[4-(1-aminocyclobutyl)phenyl]-5-phenylpyridine-3-carboxamide
|
| Synonyms |
AZD-26 AZD26 AZD 26
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~75 mg/mL (~218.39 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.28 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
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 (7.28 mM) (saturation unknown) 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. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.28 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9118 mL | 14.5590 mL | 29.1180 mL | |
| 5 mM | 0.5824 mL | 2.9118 mL | 5.8236 mL | |
| 10 mM | 0.2912 mL | 1.4559 mL | 2.9118 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
