BLU2864 is an orally bioactive, selective, ATP-competitive PRKACA inhibitor (IC50=0.3 nM). BLU2864 has anti-tumor activity and may be utilized in cancer and polycystic kidney disease study.

Physicochemical Properties

Molecular Formula	C24H19F3N4O2
Molecular Weight	452.428475618362
Exact Mass	452.146
CAS #	2810747-89-6
PubChem CID	164603354
Appearance	Off-white to light yellow solid powder
LogP	3.2
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	33
Complexity	715
Defined Atom Stereocenter Count	2
SMILES	CC1=CNC2=NC=CC(=C12)C3=NC=C(C=C3)C(=O)N[C@H]4[C@H](CC5=C4C(=CC(=C5)C(F)F)F)O
InChi Key	VRPOVDLLZCQZEG-RXVVDRJESA-N
InChi Code	InChI=1S/C24H19F3N4O2/c1-11-9-30-23-19(11)15(4-5-28-23)17-3-2-12(10-29-17)24(33)31-21-18(32)8-13-6-14(22(26)27)7-16(25)20(13)21/h2-7,9-10,18,21-22,32H,8H2,1H3,(H,28,30)(H,31,33)/t18-,21-/m0/s1
Chemical Name	N-[(1R,2S)-5-(difluoromethyl)-7-fluoro-2-hydroxy-2,3-dihydro-1H-inden-1-yl]-6-(3-methyl-1H-pyrrolo[2,3-b]pyridin-4-yl)pyridine-3-carboxamide
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)

Biological Activity

ln Vitro	In vitro, BLU2864 (40 nM and 200 nM; 5 d) suppresses cystogenesis caused by forskolin [1].
ln Vivo	In Pkd1RC/RC mice, BLU2864 (oral gavage; 45 mg/kg; once daily; 5 days) inhibits renal PKA activity [1]. In Pkd1RC/RC mice, BLU2864 (oral gavage; 30 mg/kg; once daily; 5 days) reduces PKA activity and enhances PKD [1]. FLC tumor growth is inhibited in vivo by BLU2864 (oral gavage; 30 mg/kg and 75 mg/kg; once daily; 34 days) [2].
Cell Assay	Cell Viability Assay [1] Cell Types: mIMCD3 Cell Tested Concentrations: 40 nM and 200 nM Incubation Duration: 5 days Experimental Results: Forskolin-induced in vitro cyst formation of mIMCD3 cells cultured in Matrigel was inhibited at 40 and 200 nM concentrations, respectively. 72% and 100%, respectively, relative to the control.
Animal Protocol	Animal/Disease Models: Pkd1RC/RC mice [1] Doses: 45 mg/kg Route of Administration: po (oral gavage); 45 mg/kg; one time/day; 5 days Experimental Results: Kidney basis of BLU2864-treated mice compared with control group and total PKA activity were inhibited by 74% and 87%, respectively, at 3 h, and by 46% and 56%, respectively, at 15 h. Animal/Disease Models: Pkd1RC/RC mice [1] Doses: 30 mg/kg Doses: po (oral gavage); 30 mg/kg; one time/day; 5 days Experimental Results: At 15 weeks, BLU2864-treated mice had higher urine output than Control group. demonstrated lower kidney weight, kidney volume as a percentage of body weight, and cyst index. Compared with the control group, renal basal and total PKA activities were diminished by 69% and 84%, respectively, in BLU2864-treated mice. Animal/Disease Models: FLC PDX tumor-bearing mice [2] Doses: 30 mg/kg and 75 mg/kg Route of Administration: po (oral gavage); 30 mg/kg and 75 mg/kg; one time/day; 34-day Experimental Results: Day 3 At 34 days, tumor growth was inhibited by 48.5% (P=0.003) and 45.3% (P=0.0005), respectively.
References	[1]. Protein Kinase A Downregulation Delays the Development and Progression of Polycystic Kidney Disease. J Am Soc Nephrol. 2022 Jun;33(6):1087-1104. [2]. Evaluation of PRKACA as a Therapeutic Target for Fibrolamellar Carcinoma. bioRxiv 2022.01.31.477690.

Solubility Data

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

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	2.2103 mL	11.0514 mL	22.1029 mL
	5 mM	0.4421 mL	2.2103 mL	4.4206 mL
	10 mM	0.2210 mL	1.1051 mL	2.2103 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.