BX-320 is a selective, ATP-competitive, orally bioavailable direct PDK1 inhibitor (antagonist) with IC50 of 30 nM. BX-320 also causes apoptosis. has anti-neoplastic activities.

Physicochemical Properties

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
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)

Biological Activity

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].
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].
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.
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.
References	[1]. Novel small molecule inhibitors of 3-phosphoinositide-dependent kinase-1. J Biol Chem. 2005 May 20;280(20):19867-74.

Solubility Data

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	1.8267 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.