| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| Targets |
TRPC5 (transient receptor potential cation channel subfamily C member 5). BTD is a positive modulator/activator of the TRPC5 channel, a non-selective cation channel (primarily permeable to Na+ and Ca2+). Activation of TRPC5 leads to membrane depolarization and an increase in intracellular calcium. BTD selectively activates TRPC5 over other TRP channels, making it a valuable tool for studying TRPC5-mediated functions. The EC50 for BTD on TRPC5 is 1.4 uM in a calcium flux assay. BTD also activates heteromeric channel complexes formed by TRPC5 with its relatives TRPC1 or TRPC4.
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| ln Vitro |
With EC50 values of 1.4 μM in the fluorescent microwell Ca2+ test and 1.3 μM in whole-cell patch clamp assays, BTD can activate TRPC5 [1]. With an EC50 value of 20.6 μM, BTD can activate HEK293 cells expressing TRPM8 [1]. BTD can also activate a heteromeric channel complex made up of TRPC5, TRPC1, and TRPC4, which are near cousins of TRPC5.
In cell-free assays using inside-out patches of membrane, BTD activates TRPC5 channels reconstituted in lipid bilayers, increasing the open probability (Po). The binding site is likely at the intracellular C-terminus or the voltage-sensor-like domain (VSLD). In biochemical pull-down assays, BTD can be shown to bind to the TRPC5 protein, though the exact binding pocket is not fully characterized. It has an EC50 of 1.3 uM in whole-cell patch clamp assays, confirming direct channel activation. It also has a weaker effect on TRPM8, with an EC50 of 20.6 uM in HEK293 cells expressing TRPM8, showing ~15-fold selectivity for TRPC5. |
| ln Vivo |
In cultured cells (e.g., HEK293 transfected with TRPC5), BTD (EC50 = 1.4 uM) induces a rapid influx of calcium, which can be measured with fluorescent calcium indicators. This increase in intracellular calcium triggers downstream signaling events, such as the activation of calcineurin and the translocation of nuclear factor of activated T-cells (NFAT) to the nucleus. In neurons, BTD can depolarize the cell membrane, increasing neuronal excitability. In primary dorsal root ganglion (DRG) neurons, BTD application leads to action potential firing. Its effects are blocked by TRPC5 inhibitors (e.g., HC-070 or ML204), confirming target specificity.
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| Enzyme Assay |
The activity of BTD on TRPC5 is assessed using a calcium flux assay. HEK293 cells stably or transiently expressing human TRPC5 are seeded in black-wall, clear-bottom 96-well plates. The cells are loaded with a calcium-sensitive dye, such as Fluo-4 AM, in a HEPES-buffered salt solution for 1 hour at 37degC. After washing, varying concentrations of BTD (0.01-100 uM) are added to the wells by an automated injector. Fluorescence (excitation 485 nm, emission 525 nm) is measured in real-time (1-5 minutes) using a fluorescence plate reader. The maximum fluorescence signal (peak height) is plotted against the BTD concentration to generate a concentration-response curve, and the EC50 value is calculated by nonlinear regression. For selectivity, similar assays are performed on cells expressing other TRP channels (e.g., TRPC1, TRPC4, TRPC6, TRPM8, TRPV1).
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| Cell Assay |
The standard cellular assay for BTD is the calcium flux assay described above, using HEK293 cells expressing TRPC5. Alternatively, whole-cell patch clamp electrophysiology is used to directly measure ionic currents. HEK293 cells expressing TRPC5 are voltage-clamped at -60 mV. BTD (0.1-100 uM) is applied to the extracellular solution via a rapid perfusion system. The current-voltage relationship (I-V curve) and the effect of BTD on channel gating are recorded. The EC50 for current activation is determined by measuring the current amplitude at varying concentrations. The Hill coefficient is calculated. BTD-induced currents are blocked by the TRPC5 antagonist ML204 (10 uM). For cytotoxicity, cell viability is assessed by MTT assay after 24-48 hours of BTD exposure to ensure effects are not due to toxicity.
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| Animal Protocol |
In vivo studies demonstrate that BTD can modulate pain pathways and neurodegeneration. In a rat model of painful diabetic neuropathy (induced by streptozotocin), BTD is administered intraperitoneally (i.p.) at doses of 1 and 3 mg/kg daily for 14 days. Pain behavior is assessed by measuring mechanical allodynia using von Frey filaments. BTD treatment (1 mg/kg, i.p.) significantly improves mechanical allodynia in diabetic neuropathic rats, with no effect on thermal hyperalgesia or neurological deficits. In mouse models of anxiety, TRPC5 activators like BTD are studied for their potential to reduce anxiety-like behavior. A typical study might involve administration of BTD (0.5-5 mg/kg, i.p.) 30 minutes before testing in the elevated plus maze (EPM) or open field test (OFT). BTD is not used in cancer models.
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| ADME/Pharmacokinetics |
Formal pharmacokinetic studies for BTD have not been extensively published. The compound has a molecular weight of 459.6, a LogP of 2.63, and is soluble in DMSO at up to 100 mg/mL. For in vivo studies, BTD is formulated using a vehicle such as 10% DMSO + 90% (20% SBE-beta-CD in saline), achieving a clear solution at 2.5 mg/mL (5.44 mM). In the diabetic neuropathy study, the dosing regimen was 1-3 mg/kg i.p. daily. It is not known whether BTD crosses the blood-brain barrier (BBB); however, its effects on pain behavior suggest at least peripheral activity. Its half-life in vivo is not reported. For research use, stock solutions are stored at -20degC.
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| Toxicity/Toxicokinetics |
Preclinical toxicology data for BTD are limited. In the rat diabetic neuropathy study, BTD was administered at 1 and 3 mg/kg i.p. daily for 14 days, and no significant adverse effects on body weight, general behavior, or neurological function were reported. It is not considered genotoxic or cytotoxic at low micromolar concentrations in vitro. As a TRPC5 activator, potential on-target toxicities include effects on kidney function (TRPC5 is expressed in podocytes) and the cardiovascular system. However, no specific organ toxicity has been reported. For research use, it should be handled as a potentially hazardous chemical with appropriate PPE (gloves, lab coat, goggles). It is not approved for human use.
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| References | |
| Additional Infomation |
BTD is a research-grade activator of TRPC5 channels, first described in a 2019 paper by Sadowski et al. Its chemical name is N-(3-((3r,5r,7r)-adamantan-1-yloxy)propyl)-3-(6-methyl-1,1-dioxido-2H-1,2,4-benzothiadiazin-3-yl)propanamide. The compound is an important tool for studying the physiology and pathology of TRPC5, which is implicated in brain development, pain perception, and depression. A TRPC5 activator like BTD is the opposite of a TRPC5 inhibitor (e.g., HC-070), allowing researchers to perform bidirectional modulation. BTD is not approved for therapeutic use. It is intended for laboratory research only and is typically provided as a solid powder with a purity of ≥98%. It should be stored at -20degC.
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| Molecular Formula |
C24H33N3O4S
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|---|---|
| Molecular Weight |
459.601525068283
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| Exact Mass |
459.219
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| CAS # |
896684-04-1
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| PubChem CID |
46369355
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.691
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| LogP |
2.63
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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 |
8
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| Heavy Atom Count |
32
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| Complexity |
809
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC2=C(C=C1)S(=O)(=O)N=C(N2)CCC(=O)NCCCOC34CC5CC(C3)CC(C5)C4
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| InChi Key |
YXADKMPRWFBOHW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H33N3O4S/c1-16-3-4-21-20(9-16)26-22(27-32(21,29)30)5-6-23(28)25-7-2-8-31-24-13-17-10-18(14-24)12-19(11-17)15-24/h3-4,9,17-19H,2,5-8,10-15H2,1H3,(H,25,28)(H,26,27)
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| Chemical Name |
N-[3-(1-adamantyloxy)propyl]-3-(6-methyl-1,1-dioxo-4H-1λ6,2,4-benzothiadiazin-3-yl)propanamide
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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 : ~100 mg/mL (~217.58 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.44 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 2: ≥ 2.5 mg/mL (5.44 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 | 2.1758 mL | 10.8790 mL | 21.7581 mL | |
| 5 mM | 0.4352 mL | 2.1758 mL | 4.3516 mL | |
| 10 mM | 0.2176 mL | 1.0879 mL | 2.1758 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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