BAY-3827 is a potent and specific AMPK inhibitor (antagonist) with IC50s of 1.4 nM (10 µM ATP) and 15 nM (2 mM ATP). BAY-3827 is more than 500-fold selective compared to AMPK for 331 other tested kinases. AMPK inhibits the phosphorylation of acetyl-CoA carboxylase 1 and displays potent antiproliferation activity in androgen-dependent prostate cancer/tumor cell lines.

Physicochemical Properties

Molecular Formula	C27H25FN6O
Molecular Weight	468.53
Exact Mass	468.207
CAS #	2377576-35-5
PubChem CID	134817183
Appearance	White to off-white solid powder
LogP	4.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	35
Complexity	977
Defined Atom Stereocenter Count	0
SMILES	C(NC1C2=C(NN=1)C(C)=C(F)C(C1C(C#N)=C(C)N(C)C(C)=C1C#N)=C2)(=O)C1=CC=CC=C1CC
InChi Key	OZFFKOSQNBBYCA-UHFFFAOYSA-N
InChi Code	InChI=1S/C27H25FN6O/c1-6-17-9-7-8-10-18(17)27(35)31-26-20-11-19(24(28)14(2)25(20)32-33-26)23-21(12-29)15(3)34(5)16(4)22(23)13-30/h7-11,23H,6H2,1-5H3,(H2,31,32,33,35)
Chemical Name	N-(5-(3,5-dicyano-1,2,6-trimethyl-1,4-dihydropyridin-4-yl)-6-fluoro-7-methyl-1H-indazol-3-yl)-2-ethylbenzamide
Synonyms	BAY3827 BAY-3827 BAY3827
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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	BAY-3827 (0-200 μM) has been shown to suppress AMPK kinase activity, with an IC50 value of 1.4 nM at low concentrations of 10 μM ATP and 15 nM at high concentrations of 2 mM ATP [1]. At 10 μM ATP concentration, BAY-3827 (0-200 μM) suppresses Flt3, c-Met, Rsk4, Aurora A, and IC50 values of 1324, 124, 788, and 36 nM, respectively [1]. Overnight administration of BAY-3827 significantly lowers the phosphorylation of ACC1 Ser79 in LNCaP and VCaP cells, and to a lesser degree in IMR-32, particularly Colo320 cells [1]. LNCaP and VCaP cells exhibit potent inhibitory effects when exposed to BAY-3827 (0–10 nM; 6 d) [1]. In cells implicated in the production of acylcarnitine in VCaP, BAY-3827 (1 and 5 μM; 24 and 48 hours) suppresses the expression of the LIPE gene, decreases the level of serine/threonine kinase AKT3, and limits the expression of numerous genes of the mitochondrial carnitine palmitoyltransferase (CPT) family [1]. In comparison to androgen-only treatment, BAY-3827 (5 μM; 2-4 d) significantly increases lipid droplet production [1].
Cell Assay	Cell proliferation assay[1] Cell Types: LNCaP, VCaP, 22Rv1, C4-2B, PC-3 and DU-145 prostate cancer cell lines Tested Concentrations: 0-10 nM Incubation Duration: 6 d Experimental Results: For LNCaP and DU-145 prostate Cancer cell lines demonstrated strong inhibitory effects on VCaP cells, two prostate cancer cell lines, with IC50 values of 0.28 and 1.71 nM respectively. Inhibits 22Rv1 cell proliferation with an IC50 value of 5.55 nM.
References	[1]. The potent AMPK inhibitor BAY-3827 shows strong efficacy in androgen-dependent prostate cancer models. Cell Oncol (Dordr). 2021 Jun;44(3):581-594.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~53.36 mM)

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	2.1343 mL	10.6717 mL	21.3434 mL
	5 mM	0.4269 mL	2.1343 mL	4.2687 mL
	10 mM	0.2134 mL	1.0672 mL	2.1343 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.