| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
TDAG8/GPR65[1]
T cell death-associated gene 8 (TDAG8/GPR65), a proton-sensing G protein-coupled receptor. |
|---|---|
| ln Vitro |
In HEK293 cells transiently producing hTDAG8 and mTDAG8, BTB09089 (0-18 μM; 30 min) dramatically boosted cAMP buildup, but not in normal HEK293 cells. When HEK293 cells produce hGPR4, BTB09089 does not increase the accumulation of cAMP; similarly, when HEK293 cells express hOGR1, it does not increase the accumulation of inositol phosphate [1]. At pH 7.0–7.9, but not at pH 6.5, BTB09089 (0–18 μM; 30 min) dramatically increased cAMP buildup in a dose-dependent manner [1]. BTB09089 (1–5 μM; 20 h) does not impact cell viability and dose-dependently suppresses the generation of IL-2 in the splenocytes of WT mice but not TDAG8 KO mice [1]. In LPS-stimulated ethyl sulfonate-induced peritoneal effusion cells (TG-PEC), BTB09089 (1–5 μM; 18 h) increases IL-10 production and suppresses TNF-α and IL-6 production [1].
BTB09089 is a specific agonist of TDAG8/GPR65. It induces the expression of TDAG8 and regulates the production of cytokines (e.g., IL-10, TNF-α) in T cells and macrophages. Its activity can be assessed by measuring the receptor's downstream signaling pathways, such as cAMP accumulation or MAPK activation. |
| ln Vivo |
Six hours before MCAO, rats treated with BTB09089 (5–20 μM, 8 μL; icv) have less cerebral ischemia-reperfusion injury[2].
In vivo, BTB09089 has been shown to promote neurological recovery in animal models of stroke. By modulating immune cell function, it may exert protective effects in models of neuroinflammation and tissue damage. Its role in regulating cytokine production suggests it could modulate inflammatory responses. |
| Enzyme Assay |
The agonist activity of BTB09089 on GPR65 can be assessed using cell-based functional assays. Cells expressing GPR65 are treated with the compound, and a downstream signaling event, such as the inhibition of forskolin-stimulated cAMP production, is measured. The EC50 for receptor activation can be determined from a dose-response curve.
|
| Cell Assay |
To study its effects on immune cells, primary T cells or macrophages are cultured with BTB09089. The expression of TDAG8 is measured by qPCR or Western blot. The levels of cytokines (e.g., TNF-α, IL-10, IL-6) in the culture supernatant are measured by ELISA to assess the compound's immunomodulatory effects.
|
| Animal Protocol |
Animal/Disease Models: Adult male SD rats (270-280 g), middle cerebral artery occlusion (MCAO) model[2]
Doses: 5, 10 and 20 μM, 8 μL Route of Administration: Intracerebroventricular injection, six hrs (hours) prior to MCAO Experimental Results: Up- regulated TDAG8 and Bcl-2 expression and down-regulated cleaved caspase-3 expression, while the infarction volume was decreased, and neurological deficits were ameliorated 24 and 72 h after MCAO. The in vivo efficacy of BTB09089 is often evaluated in mouse models of disease. For instance, in a model of stroke, animals are administered the compound (route and dose vary), and neurological outcomes are assessed using behavioral tests and histological analysis of brain tissue. Immune cell infiltration and cytokine levels in the brain are also measured. |
| ADME/Pharmacokinetics |
Specific ADME data for BTB09089 are not widely published. It is a small molecule (MW 387.31) that is typically dissolved in DMSO for in vitro studies. As a powder, it is stable at -20°C for up to 3 years. Its solubility and stability in vivo would require further investigation.
|
| Toxicity/Toxicokinetics |
Specific toxicological data for BTB09089 are not available in general references. As a research compound, it is not intended for human use. Standard laboratory safety precautions apply. Its safety profile would be determined through dedicated toxicology studies.
|
| References |
[1]. Onozawa Y, et al. Activation of T cell death-associated gene 8 regulates the cytokine production of T cells and macrophages in vitro. Eur J Pharmacol. 2012 May 15;683(1-3):325-31.
[2]. Ma XD, et al. TDAG8 activation attenuates cerebral ischaemia-reperfusion injury via Akt signalling in rats. Exp Neurol. 2017 Jul;293:115-123. |
| Additional Infomation |
BTB09089 is a research tool used to investigate the biological functions of the GPR65 receptor. It has been used to dissect the role of GPR65 in immune cell function, neuroinflammation, and tissue repair, with studies demonstrating its ability to modulate cytokine production in T cells and macrophages and to promote neurological recovery in animal models of stroke.
|
| Molecular Formula |
C14H12CL2N4OS2
|
|---|---|
| Molecular Weight |
387.31
|
| Exact Mass |
385.982
|
| CAS # |
245728-44-3
|
| PubChem CID |
2801217
|
| Appearance |
Yellow to orange solid powder
|
| Density |
1.6±0.1 g/cm3
|
| Boiling Point |
500.6±60.0 °C at 760 mmHg
|
| Flash Point |
256.6±32.9 °C
|
| Vapour Pressure |
0.0±1.3 mmHg at 25°C
|
| Index of Refraction |
1.733
|
| LogP |
1.82
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
23
|
| Complexity |
595
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClC1C=C(C=CC=1CSC1=NN(C)C2C=NN(C)C(C=2S1)=O)Cl
|
| InChi Key |
PIAWQVCBQLTEIY-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H12Cl2N4OS2/c1-19-11-6-17-20(2)13(21)12(11)23-14(18-19)22-7-8-3-4-9(15)5-10(8)16/h3-6H,7H2,1-2H3
|
| Chemical Name |
3-[(2,4-dichlorophenyl)methylsulfanyl]-1,6-dimethylpyridazino[4,5-e][1,3,4]thiadiazin-5-one
|
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5819 mL | 12.9096 mL | 25.8191 mL | |
| 5 mM | 0.5164 mL | 2.5819 mL | 5.1638 mL | |
| 10 mM | 0.2582 mL | 1.2910 mL | 2.5819 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
