Sabutoclax (BI-97C1), a apogossypolone derivative, is a novel and potent pan-Bcl-2 inhibitor with IC50 values of 0.31 μM, 0.32 μM, 0.20 μM and 0.62 μM for Bcl-xL, Bcl-2, Mcl-1, and Bfl-1, respectively. It demonstrated strong binding affinity to Bcl-xL in both the NMR binding assay and the ITC assay, with a Kd value of 0.11μM. In comparison to other apogossypolone derivatives, sabutoclax also demonstrated improved cell membrane permeability. Sabutoclax had an EC50 value of 0.13 μM for inhibiting cell growth in PC3 cells. Sabutoclax had an IC50 value of 0.049 μM for inducing cell apoptosis in the human BP3 cell line. Sabutoclax treatment significantly decreased the tumor size in mice receiving cancer xenografts from M2182. Sabutoclax almost completely suppressed tumor growth when administered at a dose of 5 mg/kg.

Physicochemical Properties

Molecular Formula	C42H40N2O8
Molecular Weight	700.78
Exact Mass	700.278
Elemental Analysis	C, 71.98; H, 5.75; N, 4.00; O, 18.26
CAS #	1228108-65-3
Related CAS #	1228108-65-3
PubChem CID	44224066
Appearance	Light brown to brown solid powder
Density	1.4±0.1 g/cm3
Boiling Point	905.9±65.0 °C at 760 mmHg
Flash Point	501.7±34.3 °C
Vapour Pressure	0.0±0.3 mmHg at 25°C
Index of Refraction	1.713
LogP	7.47
Hydrogen Bond Donor Count	8
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	9
Heavy Atom Count	52
Complexity	1100
Defined Atom Stereocenter Count	0
SMILES	O=C(C1=C(O)C(O)=CC2=C(O)C(C3=C(C)C=C4C(C(NCC(C5=CC=CC=C5)C)=O)=C(O)C(O)=CC4=C3O)=C(C)C=C12)NCC(C6=CC=CC=C6)C
InChi Key	RAYNZUHYMMLQQA-ZEQRLZLVSA-N
InChi Code	InChI=1S/C42H40N2O8/c1-21-15-27-29(17-31(45)39(49)35(27)41(51)43-19-23(3)25-11-7-5-8-12-25)37(47)33(21)34-22(2)16-28-30(38(34)48)18-32(46)40(50)36(28)42(52)44-20-24(4)26-13-9-6-10-14-26/h5-18,23-24,45-50H,19-20H2,1-4H3,(H,43,51)(H,44,52)/t23-,24-/m0/s1
Chemical Name	2,3,5-trihydroxy-7-methyl-N-[(2R)-2-phenylpropyl]-6-[1,6,7-trihydroxy-3-methyl-5-[[(2R)-2-phenylpropyl]carbamoyl]naphthalen-2-yl]naphthalene-1-carboxamide
Synonyms	BI-97C1; BI 97C1; BI97C1; Sabutoclax
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

Targets	Bcl-xL (Ki = 0.31 μ); BCL2 (IC50 = 0.32 μM); Mcl-1 (IC50 = 0.2 μM); Bfl-1 (IC50 = 0.62 μM)
ln Vitro	BI-97C1 potently inhibits cell growth of human prostate cancer, lung cancer, and lymphoma cell lines with EC50 values of 0.13, 0.56, and 0.049 μM, respectively, and shows little cytotoxicity against bax-/-bak-/- cells[1]. It is hypothesized that treatment with the combination of mda-7/IL-24 and BI-97C1 induces autophagy that facilitates apoptosis in conjunction with up-regulation of NOXA, accumulation of Bim, and activation of Bax and Bak[2].
ln Vivo	In addition to showing superior single-agent antitumor efficacy in a prostate cancer mouse xenograft model that relies on Mcl-1 for survival, BI-97C1 also exhibits in vivo efficacy in transgenic mice in which Bcl-2 is overexpressed in splenic B-cells[1]. Ad.5/3-mda-7 and BI-97C1 treatment significantly slows the growth of human PC xenografts in nude mice and spontaneously induced PC in Hi-myc transgenic mice. In both PC xenografts and the prostates of Hi-myc mice, tumor growth inhibition correlates with increased TUNEL staining and decreased Ki-67 expression[2].
Enzyme Assay	A Bak BH3 peptide (F-BakBH3) with the amino acid sequence GQVGRQLAIIGDDINR is purified using HPLC after being FITC-labeled at the N-terminus. For competitive binding assays, 47.5 μL of PBS (pH 7.4) and 100 nM GST-Bcl-XL ΔTM protein are preincubated at room temperature for 10 minutes with the tested substance at varying concentrations. Then, 2.5 μL of 100 nM FITC-labeled Bak BH3 peptide is added to make a final volume of 50 μL. Each assay plate contains both the wild-type and mutant Bak BH3 peptides as positive and negative controls, respectively. At excitation/emission wavelengths of 480/535 nm, following a 30-minute incubation period at room temperature, the polarization values in millipolarization units are determined using a multilabel plate reader. The experimental data are fitted to a sigmoidal dose-response nonlinear regression model to calculate the IC50. The results are the mean of three separate experiments. Bcl-2 and Mcl-1FPA function similarly. In a nutshell, 15 nM of FITC-conjugated-Bim BH3 peptide is added to 50 nM of GST-Bcl-2 or -Mcl-1 in PBS buffer after 2 minutes of incubation with various compound concentrations (4 and 11–14). After 10 minutes, the polarization of the fluorescence is measured.
Animal Protocol	1 , 3, 5 mg/kg Ethanol:Cremophor EL:saline = 10:10:80 Bcl-2 transgenic mice, human prostate cancer xenografts
References	[1]. J Med Chem . 2010 May 27;53(10):4166-76. [2]. Proc Natl Acad Sci U S A . 2011 May 24;108(21):8785-90.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~142.7 mM) Water: <1 mg/mL(slightly soluble or insoluble) Ethanol: ~54mg/mL (~77 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.75 mg/mL (3.92 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 27.5 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.4270 mL	7.1349 mL	14.2698 mL
	5 mM	0.2854 mL	1.4270 mL	2.8540 mL
	10 mM	0.1427 mL	0.7135 mL	1.4270 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.