| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
| Targets |
HIV-1 Integrase (IN) - allosteric inhibitor targeting the LEDGF/p75 binding pocket [1]
|
|---|---|
| ln Vitro |
Lavendustin B (0-1000 µM) dose-dependently reduces human erythrocytes' absorption of methylglucose, deoxyglucose, and dehydroascorbic acid in HL-60 cells; 50% inhibition is seen at about 10–30 µM. Moreover, cytochalasin B's dose-dependent binding to human erythrocyte membranes is inhibited by rising Lavendustin B concentrations [1].
Lavendustin B was identified as an inhibitor of HIV-1 integrase binding to its cellular cofactor LEDGF/p75 through structure-based virtual screening and subsequent activity assays [1]. In an AlphaScreen assay, Lavendustin B inhibited the IN-LEDGF/p75 interaction with an IC₅₀ of 94.07 μM (referenced from previous work) [1]. Docking studies revealed that the carboxylic group of Lavendustin B establishes hydrogen bond interactions with the backbone nitrogen atoms of Glu170 and His171 residues of IN, mimicking the interactions seen with LEDGF/p75 hotspot residue Asp366. Additionally, a potential hydrogen bond is formed with the hydroxyl group side chain of Thr174 [1]. The aromatic portions of Lavendustin B occupy the hydrophobic pocket within the IN dimer interface, comprising residues from both IN subunits (Thr174, Gln168, Ala169, Met178 from subunit A; Ala128, Ala129, Trp131, Trp132 from subunit B) [1]. |
| Enzyme Assay |
This study did not employ traditional enzyme activity assays on purified enzymes. Instead, it used competitive binding and transport assays to demonstrate the direct interaction of Lavendustin B with the GLUT1 transporter protein. The key assay was the measurement of D-glucose-displaceable [4-³H]cytochalasin B binding to unsealed erythrocyte ghosts. Membranes (0.38 mg protein/mL) were incubated with 10 μM cytochalasin E, 500 mM D- or L-glucose, 0.07 μCi [4-³H]cytochalasin B, and unlabeled cytochalasin B to a final concentration of 0.01 μM, along with varying concentrations of Lavendustin B. After a 10-minute incubation at room temperature, membranes were collected by centrifugation. Bound cytochalasin B was calculated from the radioactivity in the pellet. The difference in binding in the presence of D-glucose versus L-glucose represents binding to functional glucose transporters. [1]
|
| Cell Assay |
The study used cellular uptake assays to measure the functional activity of GLUT1 in the presence of Lavendustin B.
1. Cell Lines and Culture: HL-60 cells were cultured in Iscove's modified Dulbecco's medium (IMDM) with 10% fetal bovine serum and antibiotics. CHO cells (both control and GLUT1-overexpressing) were cultured in IMDM with 10% fetal bovine serum, antibiotics, and 0.25 mg/mL G418 for the transfected cells. Human erythrocytes were purified from outdated blood samples. [1] 2. Uptake Assay Protocol: For uptake assays, cells were incubated at room temperature in an incubation buffer containing radiolabeled substrates: L-[14C]ascorbic acid (for dehydroascorbic acid uptake, after oxidation with ascorbate oxidase), 2-[1,2-³H(N)]-deoxy-D-glucose, [³H]-methylglucose, or L-[3,4,5-³H(N)]-leucine, along with respective unlabeled compounds at concentrations from 0.2 to 20 mM. Uptake times varied from 10 seconds to 1 minute, with a 30-second assay used to determine initial rates of transport. Lavendustin B was added at the beginning of the assay simultaneously with the transported substrates from concentrated stock solutions (50 μM in dimethyl sulfoxide). Following uptake, cells were washed in cold phosphate-buffered saline, lysed, and incorporated radioactivity was measured by liquid scintillation counting. [1] 3. Specificity Control: Leucine transport, which is mediated by systems unrelated to glucose transporters, was measured as a control to ensure the inhibitory effects were specific to GLUT1. [1] |
| References | |
| Additional Infomation |
Lavendustin B is an aromatic amine. It is a weak tyrosine kinase inhibitor and was previously used as a negative control analog of Lavendustin A. (NCI)
Lavendustin B [5-[bis(2-hydroxybenzyl)amino]-2-hydroxybenzoic acid] is a natural product that was identified as a hit compound targeting the HIV-1 IN-LEDGF/p75 protein-protein interaction through structure-based virtual screening [1]. The compound serves as the parent molecule for the development of a new series of allosteric HIV-1 integrase inhibitors (ALLINIs) with improved potency [1]. Structural modifications were made to the Lavendustin B scaffold, including removal of the 2-hydroxy group and addition of halogen atoms (chlorine and fluorine) or methyl substituents to explore hydrophobic interactions within the binding pocket [1]. The binding mode of Lavendustin B was compared with the known ALLINI compound KF115 through superimposition studies, revealing similar interactions with key residues in the LEDGF/p75 binding pocket of HIV-1 integrase [1]. The carboxylic acid group of Lavendustin B was identified as critical for interaction with backbone nitrogen atoms of Glu170 and His171, mimicking the LEDGF/p75 hotspot residue Asp366 [1]. |
| Molecular Formula |
C21H19NO5
|
|---|---|
| Molecular Weight |
365.37926
|
| Exact Mass |
365.126
|
| CAS # |
125697-91-8
|
| PubChem CID |
3895
|
| Appearance |
White to yellow solid powder
|
| Density |
1.423 g/cm3
|
| Boiling Point |
671.1ºC at 760 mmHg
|
| Melting Point |
140-144ºC
|
| Flash Point |
359.7ºC
|
| Vapour Pressure |
6.31E-19mmHg at 25°C
|
| Index of Refraction |
1.726
|
| LogP |
3.708
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
27
|
| Complexity |
463
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
RTYOLBQXFXYMKY-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C21H19NO5/c23-18-7-3-1-5-14(18)12-22(13-15-6-2-4-8-19(15)24)16-9-10-20(25)17(11-16)21(26)27/h1-11,23-25H,12-13H2,(H,26,27)
|
| Chemical Name |
5-[bis[(2-hydroxyphenyl)methyl]amino]-2-hydroxybenzoic acid
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~136.84 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.84 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 (6.84 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7369 mL | 13.6844 mL | 27.3688 mL | |
| 5 mM | 0.5474 mL | 2.7369 mL | 5.4738 mL | |
| 10 mM | 0.2737 mL | 1.3684 mL | 2.7369 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
|
|
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
