| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
EC50: 0.96 µM (glucose-regulated protein 94, Grp94)[1][2]
Grp94 (glucose-regulated protein 94, a paralog of Hsp90). 4-Br-Bnlm selectively binds to the ATP-binding pocket of Grp94, disrupting its chaperone function. This leads to reduced misfolding and enhanced degradation of myocilin, a protein linked to hereditary open-angle glaucoma. |
|---|---|
| ln Vitro |
In human embryonic kidney (HEK) cells, 4-Br-Bnlm (0-100 µM; 24 h) suppresses the production of the 1477N mutant cardiac protein [1]. The toxicity and levels of mutant myocardin in primary human trabecular meshwork (HTM) cells are reduced by 4-Br-Bnlm (30 µM; 24 h) [1]. In a dose-dependent manner, 4-Br-Bnlm (3, 10 and 30 µM; 24 h) suppresses myocardin production in myosin cells [1]. In HTM cells, 4-Br-Bnlm (30 µM and 100 µM; 24 h) does not cause a heat shock response [2].
4-Br-Bnlm (0-100 uM; 24 h) inhibits the expression of 1477N mutant myocilin in HEK cells. At 30 uM for 24 hours, it reduces mutant myocilin levels and toxicity in primary human trabecular meshwork (HTM) cells. It also inhibits myocilin secretion in a dose-dependent manner (3, 10, and 30 uM; 24 h). |
| ln Vivo |
In the Tg-MYOCY437H mouse model, 4-Br-Bnlm (300 µM; eye drops; once daily for 12 weeks) decreased intraocular pressure and removed mutant cardiomyopathy within trabecular meshwork (TM) cells. The photonegative reaction (PhNR) defect is improved by 4-Br-Bnlm[2].
In Tg-MYOCY437H mouse glaucoma model, 4-Br-Bnlm (300 uM; eye drops; once daily for 12 weeks) clears mutant myocilin in trabecular meshwork cells and decreases intraocular pressure (IOP). It also improves photopic negative response deficits, indicating protection of retinal ganglion cell function. |
| Enzyme Assay |
The compound is dissolved in DMSO to prepare a range of concentrations (0.1 nM to 100 uM). The binding affinity to Grp94 is assessed using a fluorescence polarization (FP) competition assay with a fluorescently labeled ATP-competitive probe. Recombinant human Grp94 (25 nM) is incubated with various concentrations of 4-Br-Bnlm and 2 nM probe at room temperature for 60 minutes. FP values are measured using a microplate reader, and IC50 values are calculated by fitting to a sigmoidal dose-response curve.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: Primary human HTM cells. Tested Concentrations: 30 µM. Incubation Duration: 24 h. Experimental Results: Dramatically diminished the level of Y437H mutant myocilin without affecting WT myocilin and the RFP vector in HEK cells. Western Blot Analysis [1] Cell Types: HEK cells. Tested Concentrations: 0, 1, 3, 10, 30 and 100 µM. Incubation Duration: 24 h. Experimental Results: Dramatically diminished 1477N mutant myocilin in HEK cells expression without causing the degradation of other Hsp90-dependent clients Akt and Ras. Human embryonic kidney (HEK) cells or primary HTM cells are cultured in DMEM/10% FBS. Cells are seeded in 6-well plates at 1×10⁵ cells/well, treated with 4-Br-Bnlm (0-100 uM) for 24-48 hours, then lysed. Myocilin levels are quantified by Western blotting using anti-myocilin antibody. Cellular toxicity is assessed by LDH release assay or MTT assay. For secretion studies, culture media are collected and analyzed by ELISA. |
| Animal Protocol |
Animal/Disease Models: Tg-MYOCY437H mice[2].
Doses: 300 µM. Route of Administration: Eye drops; one time/day for 12 weeks. Experimental Results: Inhibited mutant myocilin expression and restored the PhNR deficits. Tg-MYOCY437H mice (6-8 weeks old) receive 4-Br-Bnlm (300 uM, eye drops) in the right eye once daily for 12 weeks. The left eye receives vehicle (PBS/DMSO). IOP is measured weekly using a tonometer. After the treatment period, mice are sacrificed; eyes are enucleated and sectioned for immunohistochemistry to detect myocilin clearance. Retinal function is assessed by photopic negative response (PhNR) electroretinography. |
| ADME/Pharmacokinetics |
Cannot be extracted; information not available. 4-Br-Bnlm is a research chemical; no ADME (absorption, distribution, metabolism, excretion) data has been reported. For ocular administration (eye drops), local bioavailability is the key parameter. It is expected to have low systemic exposure when administered topically. Researchers should design PK studies to characterize its half-life and tissue distribution.
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| Toxicity/Toxicokinetics |
4-Br-Bnlm has not undergone formal toxicological evaluation. In cell-based assays, it does not induce heat shock response in HTM cells at 30 uM. In the 12-week mouse study, no overt signs of ocular or systemic toxicity were reported. Standard preclinical safety assessments are not available for this research compound.
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| References |
[1]. Stothert AR, et al, Fontaine SN, Crowley VM, Mishra S, Blagg BS, Lieberman RL, Dickey CA. Exploiting the interaction between Grp94 and aggregated myocilin to treat glaucoma. Hum Mol Genet. 2014 Dec 15;23(24):6470-80.
[2]. Stothert AR, et al. Isoform-selective Hsp90 inhibition rescues model of hereditary open-angle glaucoma. Sci Rep. 2017 Dec 20;7(1):17951. |
| Additional Infomation |
4-Br-Bnlm is a Grp94-selective inhibitor with potential therapeutic application for glaucoma by reducing misfolded myocilin accumulation. It is supplied for research use only; not approved for clinical therapy. Synonyms: methyl 2-[2-[1-[(4-bromophenyl)methyl]imidazol-2-yl]ethyl]-3-chloro-4,6-dihydroxybenzoate. Molecular formula: C20H18BrClN2O4; molecular weight: 465.72.
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| Molecular Formula |
C20H18BRCLN2O4
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|---|---|
| Molecular Weight |
465.72
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| Exact Mass |
464.013
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| CAS # |
1654775-71-9
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| PubChem CID |
127049159
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
4.9
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
28
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| Complexity |
520
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC(=O)C1=C(C(=C(C=C1O)O)Cl)CCC2=NC=CN2CC3=CC=C(C=C3)Br
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| InChi Key |
FHDUIAYTJZAXNW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H18BrClN2O4/c1-28-20(27)18-14(19(22)16(26)10-15(18)25)6-7-17-23-8-9-24(17)11-12-2-4-13(21)5-3-12/h2-5,8-10,25-26H,6-7,11H2,1H3
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| Chemical Name |
methyl 2-[2-[1-[(4-bromophenyl)methyl]imidazol-2-yl]ethyl]-3-chloro-4,6-dihydroxybenzoate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: 33.33 mg/mL (71.57 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.37 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 (5.37 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 | 2.1472 mL | 10.7361 mL | 21.4721 mL | |
| 5 mM | 0.4294 mL | 2.1472 mL | 4.2944 mL | |
| 10 mM | 0.2147 mL | 1.0736 mL | 2.1472 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.
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