| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| Other Sizes |
| Targets |
BIX primarily targets the ER chaperone protein GRP78 (BiP, HSPA5), a key regulator of the unfolded protein response (UPR). Unlike broad ER stress modulators that globally activate multiple UPR branches, BIX selectively activates the ATF6 pathway to upregulate BiP/GRP78 expression. The induction of BiP by BIX is mediated by the activation of ER stress response elements (ERSEs) through the ATF6 pathway. This selective mechanism allows BIX to induce BiP in a dose-dependent manner without inducing other molecules involved in the ER stress response, such as GRP94, calreticulin, or C/EBP homologous protein (CHOP). By specifically upregulating BiP, BIX enhances the protein folding capacity of the ER, thereby attenuating ER stress and protecting cells from ER stress-induced apoptosis.
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| ln Vitro |
BiP protein is increased by BiP inducer X (5 μM; 0–12 hours; SK–N–SH cells)[1]. BiP induces the ATF6 pathway, which in turn induces BiP[1]. Cell death caused by ER stress is decreased by BiP inducer X (5 μM; 12 hours; pretreatment with SK-N-SH cells) through suppressed activation of caspases 3/7 and 4[1].
In vitro, BIX has been extensively characterized for its ability to induce BiP expression and protect cells from ER stress. In SK-N-SH neuroblastoma cells, treatment with BIX at 5 μM for 0-12 hours significantly increases BiP protein levels. The induction of BiP is mediated by the activation of ER stress response elements (ERSEs) through the ATF6 pathway. Pretreatment with BIX at 5 μM for 12 hours inhibits cell death induced by ER stress, and this protective effect involves the inhibition of caspase 3/7 and caspase 4 activation. BIX at concentrations of 1 μM and 5 μM significantly reduces tunicamycin-induced cell death, and at 5 μM, it significantly reduces tunicamycin-induced CHOP protein expression, a marker of ER stress-induced apoptosis. The level of BiP protein significantly increases 24 hours following the dose, indicating that BIX treatment produces BiP protein. BIX has also been shown to preferentially activate BiP/GRP78, with a rapid and detrimental effect on microglia and oligodendrocytes, while astrocytes and neurons are spared. |
| ln Vivo |
The use of BiP inducer X lessens the damage caused by cerebral infarction[1]. A 20 μg dose of BiP inducer -34 g (Japan SLC)[1] was administered intracerebroventricularly. The level of BiP protein significantly increased 24 hours following the dose, indicating that BIX treatment produces BiP protein in vivo.
In vivo, BIX has demonstrated significant neuroprotective effects in various animal models. In male adult ddY mice, intracerebroventricular (i.c.v.) injection of BIX at 20 μg/2 μl reduces ER stress-induced apoptosis in the penumbra region following middle cerebral artery occlusion (MCAO). BIX reduces the insults due to cerebral infarction and decreases infarct volume and brain swelling in a mouse model of cerebral ischemia. In a gerbil model of transient forebrain ischemia, BIX protects against ischemia-induced hippocampal cell death. In the mouse retina, intravitreal injection of BIX at 5 nmol significantly induces BiP protein expression. BIX also increases renal GRP78 protein levels and attenuates ischemia and reperfusion-induced renal injury in mice. In a rat model of diabetic cardiomyopathy, BIX administration for one, two, and three weeks suppressed the activation of ER stress markers and cleavage of procaspase-3. These in vivo findings suggest that BIX has the potential to be a therapeutic agent for ER stress-induced diseases, including neurodegenerative disorders and ischemic injury. |
| Enzyme Assay |
In vitro enzyme/receptor binding assays are not typically performed for BIX, as its primary mechanism of action involves the selective induction of BiP expression through the ATF6 pathway rather than direct enzyme inhibition or receptor binding. However, the compound's activity can be assessed by measuring its ability to activate ER stress response elements (ERSEs). In these assays, cells are transfected with a reporter construct containing ERSE sequences linked to a luciferase gene, and the activation of the ERSE is measured by luciferase activity following BIX treatment. Additionally, the compound's effects on the ATF6 pathway can be evaluated by assessing the cleavage and nuclear translocation of ATF6, which is a key step in the activation of this pathway. The specificity of BIX for BiP induction can be confirmed by measuring the expression of other UPR markers, such as GRP94, calreticulin, and CHOP, which are not significantly induced by BIX.
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| Cell Assay |
In vitro cell-based assays for BIX are primarily conducted using neuronal and retinal cell lines to evaluate its protective effects against ER stress-induced cell death. In these assays, cells such as SK-N-SH neuroblastoma cells are cultured in appropriate media and treated with BIX at concentrations ranging from 1 μM to 5 μM. The induction of BiP protein expression is assessed by Western blot analysis after 0-12 hours of treatment. To evaluate the protective effects of BIX, cells are pretreated with BIX at 5 μM for 12 hours, followed by exposure to ER stress inducers such as tunicamycin (2 μg/mL) or thapsigargin. Cell death is assessed using cell viability assays such as MTT or LDH release assays, and apoptosis is evaluated by measuring caspase 3/7 and caspase 4 activation. CHOP protein expression is measured by Western blot as a marker of ER stress-induced apoptosis. The effects of BIX on microglia and oligodendrocytes have also been studied in vitro, showing a rapid and detrimental effect on these cell types.
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| Animal Protocol |
In vivo animal experiments for BIX are conducted in various rodent models to evaluate its neuroprotective and renoprotective effects. In the mouse model of cerebral ischemia induced by middle cerebral artery occlusion (MCAO), BIX is administered via intracerebroventricular (i.c.v.) injection at a dose of 20 μg/2 μl. The compound's effects on infarct volume and brain swelling are assessed, and ER stress-induced apoptosis in the penumbra region is evaluated. In the gerbil model of transient forebrain ischemia, BIX is administered to evaluate its protective effects against hippocampal cell death. In the mouse retina, BIX is administered via intravitreal injection at 5 nmol, and BiP protein expression is assessed by Western blot or immunohistochemistry. In a rat model of diabetic cardiomyopathy, BIX is administered for one, two, and three weeks, and the expression of ER stress markers and apoptotic markers is assessed by immunoblotting. In a mouse model of renal ischemia-reperfusion injury, BIX administration increases renal GRP78 protein levels and attenuates renal injury. These in vivo studies demonstrate the potential of BIX as a therapeutic agent for ER stress-related diseases.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic (PK) data for BIX are limited, as the compound is primarily a research tool and has not undergone extensive preclinical or clinical development. However, the compound's physicochemical properties provide some insights into its potential PK profile. BIX has a molecular weight of 209.22 g/mol and is soluble in DMSO. For in vivo formulations, BIX can be prepared in 10% DMSO + 90% saline at a concentration of 2 mg/mL (9.56 mM). The compound is typically administered via intracerebroventricular (i.c.v.) injection or intravitreal injection in animal studies, which bypasses first-pass metabolism and allows direct delivery to the target tissue. The compound's stability in biological fluids and its potential for protein binding are unknown. The lack of comprehensive PK data underscores the need for further studies to characterize BIX's absorption, distribution, metabolism, and excretion (ADME) properties if it is to be considered for clinical development.
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| Toxicity/Toxicokinetics |
The toxicity profile of BIX is primarily derived from its hazard classification for laboratory handling. BIX is classified as very toxic if swallowed (R28), irritating to skin (R38), with a risk of serious damage to eyes (R41), and toxic with danger of serious damage to health by prolonged exposure (R48). These classifications indicate that BIX should be handled with extreme care in the laboratory, using appropriate personal protective equipment and working in a well-ventilated area. In vitro studies have shown that BIX has a rapid and detrimental effect on microglia and oligodendrocytes, while astrocytes and neurons are spared. This selective toxicity suggests that BIX may have cell type-specific effects that could be relevant for its therapeutic potential. The compound's safety in humans has not been studied, and it is not intended for human use. Further toxicological studies are needed to fully characterize the safety profile of BIX.
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| References |
[1]. Kudo T, et al. A molecular chaperone inducer protects neurons from ER stress. Cell Death Differ. 2008;15(2):364-375.
[2]. Yoo SA, et al. A novel pathogenic role of the ER chaperone GRP78/BiP in rheumatoid arthritis. J Exp Med. 2012;209(4):871-886. |
| Additional Infomation |
2-(3,4-Dihydroxyphenyl)-2-oxoethyl thiocyanate belongs to the thiocyanate class of compounds. Its structure is similar to 3,4-dihydroxyacetophenone, except that one methyl hydrogen atom is replaced by a sulfur atom in a cyanothiodiyl (-SC#N) group. Studies have found that this compound can induce the expression of the endoplasmic reticulum molecular chaperone protein GRP78 (a 78 kDa glucose-regulated protein, BiP, a highly conserved member of the 70 kDa heat shock protein family), thereby attenuating the unfolded protein response. It can protect neurons and retinal cells from endoplasmic reticulum stress-induced cell death. This compound belongs to the thiocyanate, aromatic ketone, and catechol classes of compounds.
BIX (BiP Inducer X) is a selective inducer of immunoglobulin heavy chain binding protein (BiP)/GRP78 and an ER chaperone inducer. It was identified through a high-throughput screening (HTS) of a library of 10,000 compounds, with BIX being the compound with the highest BiP-inducing activity. BIX induces BiP expression in a dose-dependent manner without inducing other molecules involved in the ER stress response. The induction of BiP by BIX is mediated by activation of ER stress response elements (ERSEs) through the ATF6 pathway. BIX protects against ER-stress induced cell death in neuronal and retinal cell lines and protects against ischemia-induced hippocampal cell death in vivo. BIX has been shown to extend recombinant CHO cell culture longevity and increase monoclonal antibody productivity when co-supplemented with DMSO. The compound is supplied as a powder with purity ≥98% (HPLC) and is soluble in DMSO. BIX is for research use only and is not for human use. It has not yet progressed to clinical trials or received regulatory approval for any indication. The compound remains a valuable research tool for studying ER stress, the unfolded protein response, and the potential therapeutic applications of BiP induction in neurodegenerative diseases, ischemic injury, and other ER stress-related disorders. |
| Molecular Formula |
C9H7NO3S
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| Molecular Weight |
209.22178
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| Exact Mass |
209.014
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| CAS # |
101714-41-4
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| PubChem CID |
16656807
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| Appearance |
Brown to gray solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
517.1±45.0 °C at 760 mmHg
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| Melting Point |
127 °C
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| Flash Point |
266.5±28.7 °C
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| Vapour Pressure |
0.0±1.4 mmHg at 25°C
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| Index of Refraction |
1.665
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| LogP |
1.53
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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 |
3
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| Heavy Atom Count |
14
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| Complexity |
260
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N#CSCC(C1C=CC(O)=C(O)C=1)=O
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| InChi Key |
SVFLBLCWKKQKDW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H7NO3S/c10-5-14-4-9(13)6-1-2-7(11)8(12)3-6/h1-3,11-12H,4H2
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| Chemical Name |
[2-(3,4-dihydroxyphenyl)-2-oxoethyl] thiocyanate
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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 |
| 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 : ~50 mg/mL (~238.98 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (11.95 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 | 4.7797 mL | 23.8983 mL | 47.7966 mL | |
| 5 mM | 0.9559 mL | 4.7797 mL | 9.5593 mL | |
| 10 mM | 0.4780 mL | 2.3898 mL | 4.7797 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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