B026 is a selective and orally bioactive p300/CBP histone acetyltransferase (p300/CBP HAT) inhibitor (antagonist) with IC50s of 1.8 nM and 9.5 nM for p300 and CBP. B026 has anti-cancer effect against androgen receptor-positive (AR+) prostate cancer/tumor cells.

Physicochemical Properties

Molecular Formula	C27H23F4N5O4
Molecular Weight	557.496239900589
Exact Mass	557.168
CAS #	2379416-48-3
PubChem CID	146205579
Appearance	Typically exists as solid at room temperature
LogP	3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	3
Heavy Atom Count	40
Complexity	1030
Defined Atom Stereocenter Count	2
SMILES	CN1C=C(C=N1)C2=CC3=C(C=C2)[C@@]4(CC3)C(=O)N(C(=O)N4)CC(=O)N5CC6=C(C=CC(=C6)F)OC[C@H]5C(F)(F)F
InChi Key	VDEZYUOBIXKTHI-NVQXNPDNSA-N
InChi Code	InChI=1S/C27H23F4N5O4/c1-34-11-18(10-32-34)15-2-4-20-16(8-15)6-7-26(20)24(38)36(25(39)33-26)13-23(37)35-12-17-9-19(28)3-5-21(17)40-14-22(35)27(29,30)31/h2-5,8-11,22H,6-7,12-14H2,1H3,(H,33,39)/t22-,26-/m0/s1
Chemical Name	(3S)-3'-[2-[(3S)-7-fluoro-3-(trifluoromethyl)-3,5-dihydro-2H-1,4-benzoxazepin-4-yl]-2-oxoethyl]-6-(1-methylpyrazol-4-yl)spiro[1,2-dihydroindene-3,5'-imidazolidine]-2',4'-dione
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	B026 exhibits antiproliferative action against cancer cell lines (0-5 μM; 12 hours) [1]. B026 (0–5 μM; 6 hours; MV-4–11 cells) suppresses the expression of H3K27Ac [1]. B026 (0-5 μM; 12 hours; MV-4-11 cells) acts on the cellular p300 protein and, in a dose-dependent way, raises the protein's thermal stability [1]. The expression of MYC, a crucial oncogenic transcription factor controlled by a super-enhancer region containing p300/CBP, is decreased by B026 (0-1 μM; 24 hours; MV-4-11 cells) [1].
ln Vivo	B026 (1-3 mg/kg; IV and PO; 0-24 hours; male SD rats) has good oral exposure (AUC=3.71 μM·h) and good oral bioavailability (F=56%) in rats, but low clearance (13.4 mL/min/kg) [1]. In a dose-dependent manner, B026 (50-100 mg/kg; oral; daily for 28 days; balb/c female mice) inhibits the growth of tumors [1].
Cell Assay	Cell Viability Assay[1] Cell Types: Maver-1, MV-4-11, K562, Kasumi-1, LnCaP-FGC and 22Rv1 Cell Tested Concentrations: 0-5 μM Incubation Duration: 12 hrs (hours) Experimental Results: Inhibition of cell growth, IC50 value 2.6, 4.2, 4.4, 9.8, 40.5 and 104.4 nM for Maver-1, MV-4-11, 22Rv1, LnCaP-FGC, Kasumi-1 and K562 cells, respectively. Western Blot Analysis [1] Cell Types: MV-4-11 cells Tested Concentrations: 0, 0.313, 0.625, 1.25, 2.5 and 5 μM Incubation Duration: 6 hrs (hours) Experimental Results: The expression of H3K27Ac diminished in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: Balb/c female mice with MV-4-11 xenografts [1] Doses: 50 and 100 mg/kg Route of Administration: Oral; Route of Administration: Oral. one time/day for 28 days. Experimental Results: Inhibition of tumor growth, TGI was 75.0% at 50 mg/kg and 85.7% at 100 mg/kg. Animal/Disease Models: Male SD rat [1] Doses: 1 and 3 mg/kg Route of Administration: intravenous (iv) (iv)injection and oral administration; 0.25, 0.5, 1, 2, 4, 8 and 24 hrs (hrs (hours)) Experimental Results: 1.19 intravenous (iv) (iv)injection (1 mg/ kg) Oral (3 mg/kg) T1/2 (h) 1.5 1.02 Tmax (h) 0.83 Cmax (μM) 1.12 AUClast (μM·h) 2.22 3.71 Vdss (L/kg) 1.5 CL_obs (mL/min/kg) 13.4 PPB % 98.2 F % 56
References	[1]. Discovery of Highly Potent, Selective, and Orally Efficacious p300/CBP Histone Acetyltransferases Inhibitors. J Med Chem. 2020 Feb 13;63(3):1337-1360.

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	1.7937 mL	8.9686 mL	17.9372 mL
	5 mM	0.3587 mL	1.7937 mL	3.5874 mL
	10 mM	0.1794 mL	0.8969 mL	1.7937 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.