Bractoppin is a potent and selective inhibitor of phosphopeptide recognition by the BRCA1 tBRCT domain, which selectively interrupts BRCA1 tBRCT-dependent signals evoked by DNA damage.

Physicochemical Properties

Molecular Formula	C25H23FN4O
Molecular Weight	414.4747
Exact Mass	414.19
Elemental Analysis	C, 72.45; H, 5.59; F, 4.58; N, 13.52; O, 3.86
CAS #	2290527-07-8
PubChem CID	137519541
Appearance	White to off-white solid powder
Density	1.299±0.06 g/cm3
Boiling Point	625.7±65.0 °C
LogP	4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	31
Complexity	604
Defined Atom Stereocenter Count	0
SMILES	FC1=C([H])C([H])=C([H])C([H])=C1C([H])([H])N1C([H])([H])C([H])([H])N(C(C2C([H])=C([H])C3=C(C=2[H])N([H])C(C2C([H])=C([H])C([H])=C([H])C=2[H])=N3)=O)C([H])([H])C1([H])[H]
InChi Key	UHDGSNOZRAAGIZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H23FN4O/c26-21-9-5-4-8-20(21)17-29-12-14-30(15-13-29)25(31)19-10-11-22-23(16-19)28-24(27-22)18-6-2-1-3-7-18/h1-11,16H,12-15,17H2,(H,27,28)
Chemical Name	[4-[(2-fluorophenyl)methyl]piperazin-1-yl]-(2-phenyl-3H-benzimidazol-5-yl)methanone
Synonyms	Bractoppin
HS Tariff Code	2934.99.03.00
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

Bractoppin showed nanomolar effectiveness in the MST experiment, with a binding IC50 value of 74 nM, in distancing the homologous BACH1 phosphopeptide substrate of BRCA1 tBRCT [1]. ECT2, TOPBP1 1/2, TOBP1 7/8, and Bractoppin (0-100 μM) did not bind to fluorescently tagged tBRCT domains in the antagonistic profile of the competitive MST assay[1]. Whereas bractoppin (100 μM) has minimal influence on the accumulation of irradiation MDC1 at sites of DNA damage, it interferes locally with the development of irradiated BRCA1-containing foci. Similarly, on irradiated TOPBP1 (a numerous physically similar tBRCT), bractoppin prevents G2 arrest generated by 4 Gy radiation in a dose-dependent manner. Recruitment is not significantly affected by bractoppin (10–100 μM; 0.5 h prior to irradiation (32)) or domain-specific proteins (10 μM, 30 μM) [1]. The percentages of cells arrested in the G2 phase were 64%, 42%, and 25% at 100 μM, respectively[1].

References

[1]. Targeting Phosphopeptide Recognition by the Human BRCA1 Tandem BRCT Domain to Interrupt BRCA1-Dependent Signaling. Cell Chem Biol. 2018 Jun 21;25(6):677-690.e12.

Solubility Data

Solubility (In Vitro)

DMSO: 83~250 mg/mL (200.3~603.2 mM) Ethanol: ~83 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.02 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (5.02 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (5.02 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4127 mL	12.0636 mL	24.1272 mL
	5 mM	0.4825 mL	2.4127 mL	4.8254 mL
	10 mM	0.2413 mL	1.2064 mL	2.4127 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.