| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
PDK1 (3-phosphoinositide-dependent protein kinase 1). BX-320 is a selective, ATP-competitive, orally active inhibitor of PDK1 with an IC50 of 30 nM. It is selective for PDK1 over a panel of 10 additional kinases, where it shows IC50s >820 nM. PDK1 is a key activator of AKT (via phosphorylation of Thr308) and is central to the PI3K/AKT/mTOR signaling pathway.
|
|---|---|
| ln Vitro |
BX-320 attaches itself to PDK1's ATP binding site. Additionally, BX-320 inhibits CDK2b/cyclin E, Chck1, c-Kit, KDR, PKA, GSK3β, and PKC with IC50 values of 0.82, 0.89, 1.4, 1.5, 4.0, and 5.7 μM, in that order[1]. In addition to inhibiting the anchorage-dependent development of several tumor cell lines in culture or inducing apoptosis, BX-320 disrupts PDK1/Akt signaling in tumor cells[1]. BX-320 causes apoptosis and suppresses the development of MDA-468 breast cancer cells (IC50=0.6 μM). After 48 hours of treatment, BX-320 causes a 12-fold increase of caspase-3/7 activity (IC50=0.5 μm), suggesting a potent proapoptotic response[1]. For 18 hours, BX-320 (0.3–10 μM) significantly lowers the levels of p-Thr308-Akt and p-Thr386-S6K1[1].
In vitro, BX-320 (0.1-10 uM) inhibits PDK1 kinase activity, leading to a reduction in the activating phosphorylation of AKT on Thr308. This results in the suppression of downstream anti-apoptotic signals and the induction of apoptosis (programmed cell death). The compound has an IC50 of 30 nM for PDK1 inhibition and demonstrates selectivity over other related kinases. |
| ln Vivo |
Blood-borne metastases model: BX-320 (oral dose of 200 mg/kg, twice day for 21 days) is effective. Following the injection of tumor cells into the tail vein, BX-320 suppresses the formation of LOX melanoma tumors in the lungs of naked mice. BX-320 is effective in an in vivo tumor model, which may indicate that it inhibits the growth of the tumor once it has been implanted in the lung or the productive implantation of tumor cells[1].
In vivo, BX-320 inhibits the growth of lung tumors in a LOX melanoma mouse model of blood-borne metastasis when administered at 200 mg/kg. It is orally active and has been shown to induce apoptosis and reduce tumor burden in preclinical models. This demonstrates its potential as a cancer therapeutic by targeting the master PDK1/AKT signaling node. |
| Enzyme Assay |
Standard cell‑free assays for BX-320 are performed to measure its inhibition of PDK1 kinase activity. Recombinant human PDK1 enzyme (5-10 ng) is incubated with an AKT peptide substrate and varying concentrations of BX-320 (0.01-10000 nM) in a reaction buffer (e.g., 50 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT) containing 10 uM ATP. The reaction proceeds for 30-60 minutes at 22degC. The amount of phosphorylated peptide product is measured using a luminescence-based ADP-Glo™ assay, an ELISA kit, or a time-resolved FRET (TR-FRET) assay. The IC50 is calculated from the resulting inhibition curve and should be approximately 30 nM. For selectivity assays, the compound is screened against a panel of other AGC kinases (e.g., PKA, PKC, SGK, AKT) using the same assay format.
|
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: MDA-468 breast cancer cells Tested Concentrations: 31.6 nM, 100 nM, 316.22 nM, 1 μM, 3.162 μM, 10 μM, and 31.6 μM Incubation Duration: 72 hrs (hours) Experimental Results: Blocked the growth of MDA-468 cells (IC50=0.6 μM), which are PTEN -negative breast tumor cells expressing high levels of activated Akt. Western Blot Analysis[1] Cell Types: PC-3 cells Tested Concentrations: 0, 0.3, 1, 3, 10 μM Incubation Duration: 18 hrs (hours) Experimental Results: diminished the amount of both phospho -Thr308-Akt and phospho-Thr386-S6K1. For cellular assays, cancer cell lines (e.g., LOX melanoma or A549 lung cancer cells) are seeded in 96‑well plates (5,000 cells/well) in DMEM/10% FBS. After 24 hours, cells are treated with varying concentrations of BX-320 (0.1-10000 nM) for 48-72 hours. Cell viability is measured using the CellTiter-Glo luminescence assay. To assess pathway inhibition, cells are treated with BX-320 for 2-6 hours, and cell lysates are analyzed by Western blot. Blots are probed with antibodies against p-AKT (Thr308, direct substrate of PDK1) and p-AKT (Ser473, downstream of mTORC2). A selective reduction in p-AKT (Thr308) indicates on‑target PDK1 inhibition. Apoptosis is assessed by flow cytometry using Annexin V/PI staining after 24 hours of treatment. |
| Animal Protocol |
Animal/Disease Models: Athymic (nu/nu ) female mice, 6-8 weeks old[1]
Doses: 200 mg/kg; dose volume was 10 mL/kg Route of Administration: po (oral gavage) twice (two times) daily (12 h apart) Experimental Results: Dramatically inhibited the growth of lung tumors in this model. In vivo efficacy studies are performed in a mouse model of metastatic melanoma. Female athymic nude mice (6-8 weeks, 20-25 g) are injected intravenously with 5×10⁵ LOX melanoma cells to induce lung metastases. From day 1 to day 21 post-injection, mice are treated orally (by gavage) with BX-320 at a dose of 200 mg/kg suspended in a vehicle such as 0.5% methylcellulose. The dosing frequency is once daily. Control mice receive vehicle. At the end of the study (day 21), mice are euthanized, and their lungs are harvested. Lung tumor burden (number and size of metastatic nodules on the lung surface) is assessed under a dissecting microscope. For a subcutaneous xenograft model, mice are injected subcutaneously with A549 cells. When tumors reach 100 mm3, mice are treated orally with BX-320 (100-200 mg/kg daily) for 2-3 weeks. Tumor volume is measured by calipers. Body weight is monitored to assess tolerability. |
| ADME/Pharmacokinetics |
Toxicity data are limited. BX-320 is an oral inhibitor of PDK1. At the effective dose of 200 mg/kg in mice, it was well-tolerated. No specific off-target toxicity has been widely reported, but standard side effects of the PDK1/AKT pathway inhibition (e.g., metabolic effects) could occur.
|
| Toxicity/Toxicokinetics |
BX-320 is a selective, ATP-competitive, oral inhibitor of PDK1 with an IC50 of 30 nM. It has been used to study the role of the PI3K/PDK1/AKT pathway in cancer and other diseases. CAS: 702676-93-5.
|
| References |
| Molecular Formula |
C23H31BRN8O3
|
|---|---|
| Molecular Weight |
547.45
|
| Exact Mass |
546.17
|
| CAS # |
702676-93-5
|
| PubChem CID |
657138
|
| Appearance |
White to off-white solid powder
|
| Density |
1.484g/cm3
|
| Index of Refraction |
1.673
|
| LogP |
4.476
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
35
|
| Complexity |
734
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(C)(C(=O)N)C(=O)NCCCNC1=NC(=NC=C1Br)NC2=CC(=CC=C2)NC(=O)N3CCCC3
|
| InChi Key |
ZNSULAZTNWFKEW-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C23H31BrN8O3/c1-23(2,19(25)33)20(34)27-10-6-9-26-18-17(24)14-28-21(31-18)29-15-7-5-8-16(13-15)30-22(35)32-11-3-4-12-32/h5,7-8,13-14H,3-4,6,9-12H2,1-2H3,(H2,25,33)(H,27,34)(H,30,35)(H2,26,28,29,31)
|
| Chemical Name |
N'-[3-[[5-bromo-2-[3-(pyrrolidine-1-carbonylamino)anilino]pyrimidin-4-yl]amino]propyl]-2,2-dimethylpropanediamide
|
| 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: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 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 | 1.8267 mL | 9.1333 mL | 18.2665 mL | |
| 5 mM | 0.3653 mL | 1.8267 mL | 3.6533 mL | |
| 10 mM | 0.1827 mL | 0.9133 mL | 1.8267 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
