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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
TREK-1 channel (TWIK-related potassium channel-1). Spadin TFA is a potent and selective TREK-1 channel blocker. By inhibiting this leak potassium channel, it prevents potassium efflux, leading to the depolarization of serotonergic neurons in the dorsal raphe nucleus. This enhances 5-HT neurotransmission, mimicking the effects of selective serotonin reuptake inhibitors (SSRIs).
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| ln Vitro |
The spadin inhibitory effect on TREK-1 channels is exhibited by spadin TFA (100 nM; COS-7 cells), which also blocks 63% of TREK-1 currents induced by arachidonic acid [1]. In wild-type mouse brain slices, spadin TFA (100 nM) inhibits TREK-1 channel activity in CA3 hippocampal neurons [1].
In vitro, Spadin TFA (100 nM) has an inhibitory effect on the TREK-1 channel expressed in COS-7 cells, blocking 63% of the TREK-1 current stimulated by arachidonic acid. This blockade is specific, and the compound has no significant effect on other potassium channels, making it a highly selective pharmacological tool for studying TREK-1 function in isolated cells and brain slices. |
| ln Vivo |
In the dorsal raphe nucleus (DRN), spadin TFA (10 μM, 100 μL; i.p.; 30 min; male C57Bl/6J and TREK-1-deficient mice) enhances the rate at which 5-HT neurons fire [2]. The antidepressant effects of spadin TFA (0.01-100 μM, 100 μL; intracerebroventricular, intraperitoneal, and ICV; daily for 7 days; male C57Bl/6J and TREK-1-deficient mice) have been observed in mice [2].
In vivo, Spadin TFA demonstrates potent antidepressant-like activity in animal models. It enhances dorsal raphe nucleus 5-HT neurotransmission and induces hippocampal CREB activation and neurogenesis. These effects are consistent with known antidepressant mechanisms. Spadin TFA produces a robust and fast behavioral response in forced swim and tail suspension tests, comparable to classical antidepressants but with a distinct mechanism of action. |
| Enzyme Assay |
A non-cell-based electrophysiological assay is used. TREK-1 channels are expressed in Xenopus oocytes or CHO cells. Whole-cell patch-clamp recordings are performed. Cells are held at a potential (e.g., -80 mV), and currents are evoked by voltage steps. Spadin TFA (1-1000 nM) is applied to the bath solution. The percentage of channel current blocked is calculated. The IC50 is determined by fitting a Hill equation to the inhibition curve.
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| Cell Assay |
Cell lines (e.g., COS-7) are transfected with a TREK-1 expressing plasmid. After 48 hours, cells are loaded with the voltage-sensitive dye DiBAC4(3). Cells are treated with Spadin TFA (0-1000 nM) in the presence of arachidonic acid (to activate TREK-1). The change in fluorescence intensity, which correlates with membrane depolarization, is measured by a plate reader. This fluorescence-based assay is a higher-throughput alternative to patch clamping.
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| Animal Protocol |
Animal/Disease Models: Male C57Bl/6J and TREK-1 deficient mice[2]
Doses: 10 μM Route of Administration: intraperitoneal (ip)injection; for 30 min Experimental Results: Increased of the 5-HT neuron firing rate in the dorsal raphe nucleus (DRN). Animal/Disease Models: Male C57Bl/6J and TREK-1 deficient mice[2] Doses: 0.01-100 μM Route of Administration: Intracerebroventrical injection (100 nM), intraperitoneal (ip)injection (1-100 μM) and intravenous (iv) injection (0.01-1 μM); daily, for 7 days Experimental Results: Had any effect on mouse locomotion analyzed in short- or long-time after the drug injection. For the forced swim test, male Swiss mice (25-30 g) are treated with Spadin TFA (0.1-10 mg/kg, i.p.). 30 min later, they are placed individually in a cylinder of water for 6 min. The duration of immobility is recorded during the final 4 min of the test. A reduction in immobility time indicates an antidepressant-like effect. For a neurogenesis study, mice are treated daily for 14 days, and BrdU is injected to label dividing cells in the hippocampus. |
| ADME/Pharmacokinetics |
PK data for Spadin TFA is limited. As a peptide (MW 2126.36 kDa), it has a short plasma half-life and is typically administered via intraperitoneal or intravenous injection. It has poor oral bioavailability. Systemic administration in mice (0.1-10 mg/kg, i.p.) leads to rapid absorption and CNS penetration, with behavioral effects observed within 30-60 minutes post-injection.
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| Toxicity/Toxicokinetics |
Spadin TFA is a research reagent and not for human therapeutic use. In acute toxicity studies in rodents, Spadin TFA is well-tolerated at doses up to 10 mg/kg (i.p.) with no gross behavioral or neurological deficits reported. No significant effects on body weight or general health were observed. High doses may cause off-target effects due to its membrane depolarizing activity. Standard safety precautions for peptide handling should be used.
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| References |
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| Additional Infomation |
Spadin TFA is a potent research tool for investigating the role of TREK-1 channels in depression and neurophysiology. It demonstrates that blocking TREK-1 is a viable strategy for achieving antidepressant effects, distinct from traditional SSRIs. This compound is not FDA-approved and has not completed clinical trials. It should be stored at -20degC to -80degC, protected from light, and freeze-thaw cycles should be avoided.
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| Molecular Formula |
C92H143F3N26O24
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| Molecular Weight |
2126.36
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| Related CAS # |
Spadin;1270083-24-3
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| Appearance |
White to off-white solid powder
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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) |
H2O :~25 mg/mL (~11.76 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (1.18 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 (1.18 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (1.18 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 50 mg/mL (23.51 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.4703 mL | 2.3514 mL | 4.7029 mL | |
| 5 mM | 0.0941 mL | 0.4703 mL | 0.9406 mL | |
| 10 mM | 0.0470 mL | 0.2351 mL | 0.4703 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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