(±)-BI-D is a novel and potent allosteric/noncatalytic site integrase inhibitor that binds integrase at the LEDGF/p75 binding site. It binds with higher affinity to a tetrameric form compared to a dimeric form of integrase. The binding of integrase (IN) to lens epithelium-derived growth factor (LEDGF)/p75 in large part determines the efficiency and specificity of HIV-1 integration. However, a significant residual preference for integration into active genes persists in Psip1 (the gene that encodes for LEDGF/p75) knockout (KO) cells.

Physicochemical Properties

Molecular Formula	C25H27NO4
Molecular Weight	405.4862
Exact Mass	405.194
CAS #	1416258-16-6
PubChem CID	71592850
Appearance	Light yellow to yellow solid powder
Density	1.2±0.1 g/cm3
Boiling Point	587.3±50.0 °C at 760 mmHg
Flash Point	309.0±30.1 °C
Vapour Pressure	0.0±1.7 mmHg at 25°C
Index of Refraction	1.610
LogP	5.28
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	30
Complexity	602
Defined Atom Stereocenter Count	0
InChi Key	ZFERZAMPQIXCPM-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H27NO4/c1-15-21(23(24(27)28)30-25(2,3)4)22(18-9-5-6-10-19(18)26-15)17-11-12-20-16(14-17)8-7-13-29-20/h5-6,9-12,14,23H,7-8,13H2,1-4H3,(H,27,28)
Chemical Name	2-[4-(3,4-dihydro-2H-chromen-6-yl)-2-methylquinolin-3-yl]-2-[(2-methylpropan-2-yl)oxy]acetic acid
Synonyms	(±)-BI-D; (±)-BI D
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

BI-D, a potent ALLINI that binds IN at the LEDGF/p75 binding site (28), was synthesized as described in Supplementary Methods. E9 MEF cells (5 x 103) seeded in wells of a 96-well plate were infected in quadruplicate with HIV-Luc (5 x 105 RT-cpm) in the presence of serial dilutions of BI-D or dimethyl sulfoxide solvent control to determine half-maximal inhibitory concentration (IC50) values of the drug. [1] The IC50 of the BI-D inhibitor was determined during the acute phase of HIV-1 infection, at the time LEDGF/p75 and HRP2 are known to function. Approximately 2.4–2.9 µM of BI-D was required to inhibit 50% of HIV-Luc infection of WT and Hdgfrp2 KO cells, while the IC50 decreased dramatically, to 160–200 nM, in Psip1 and double-KO cells. In contrast, the IC50 value for the IN active site inhibitor raltegravir was similar in each cell type.[1]

References

[1]. HRP2 determines the efficiency and specificity of HIV-1 integration in LEDGF/p75 knockout cells but does not contribute to the antiviral activity of a potent LEDGF/p75-binding site integrase inhibitor. Nucleic Acids Res. 2012 Dec;40(22):11518-30. [2]. Minimizing the Contribution of Enterohepatic Recirculation to Clearance in Rat for the NCINI Class of Inhibitors of HIV. ACS Med Chem Lett. 2014 Apr 16;5(6):711-6.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 100 mg/mL (~246.62 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.17 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 25.0 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: ≥ 2.5 mg/mL (6.17 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 25.0 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: ≥ 2.5 mg/mL (6.17 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 25.0 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	2.4662 mL	12.3308 mL	24.6615 mL
	5 mM	0.4932 mL	2.4662 mL	4.9323 mL
	10 mM	0.2466 mL	1.2331 mL	2.4662 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.