ROS production; Bacterial; Apoptosis
Ecabet Sodium inhibits urease activity of H. pylori.
It may inhibit the adhesion of H. pylori to gastric epithelial cells.
It may have anti‑ATPase function involved in bactericidal effects in the acidic stomach environment. [1]

Ecabet Sodium can inhibit the adhesion of H. pylori to gastric epithelial cells, which is the initial step in H. pylori infection.
It can inhibit the urease activity of H. pylori. The anti‑H. pylori action induced by ecabet sodium is dependent on protons.
It may have anti‑ATPase function that is considered to be involved in bactericidal effects in the acidic environment of the stomach. [1]

Pre-treatment with ecabet sodium at concentrations of 5, 10, 20, 40, and 80 μg/mL for 1 hour followed by 125 μM neomycin for 1 hour in zebrafish larvae (5-7 dpf) protected against neomycin-induced hair cell loss in a dose-dependent manner. The mean number of viable hair cells in four neuromasts (SO1, SO2, O1, OC1) increased with ecabet sodium pre-treatment. At 40 μg/mL ecabet sodium, the mean hair cell count was 12.98 ± 2.59 cells, which was not significantly different from the negative control group (14.15 ± 1.39 cells, p = 0.72) and significantly higher than the positive control (neomycin only) group (7.45 ± 0.91 cells, p < 0.01) [2].
Ecabet sodium (40 μg/mL) pre-treatment significantly reduced neomycin-induced apoptosis. TUNEL assay showed that neomycin alone gave a mean TUNEL score of 0.48 ± 0.60 (many TUNEL-positive cells), whereas pre-treatment with 40 μg/mL ecabet sodium significantly increased the TUNEL score to 1.58 ± 0.72 (p < 0.01), indicating fewer apoptotic hair cells [2].
Ecabet sodium inhibited neomycin-induced reactive oxygen species (ROS) generation. Neomycin (125 μM) treatment increased ROS production to 183% of the negative control level. Pre-treatment with 40 μg/mL ecabet sodium reduced ROS production to 105% of the negative control level (p = 0.04 compared to neomycin alone; p = 0.99 compared to negative control) [2].
Ecabet sodium protected mitochondria in hair cells. DASPEI staining showed that neomycin alone decreased the mean DASPEI score to 0.33 ± 0.43 (reduced viable mitochondria), while pre-treatment with 40 μg/mL ecabet sodium increased the DASPEI score to 1.30 ± 0.26 (p < 0.01 compared to neomycin alone) [2].
Scanning electron microscopy revealed that ecabet sodium (40 μg/mL) pre-treatment substantially restored the destruction of kinocilium and stereocilia bundles in neuromasts caused by 125 μM neomycin [2].

Wild-type and transgenic zebrafish larvae (Brm3C: EGFP) were produced by mating adult fish reared at 28.5 ± 1°C in a zebrafish facility. Larvae were maintained in embryo medium (15 mM NaCl, 0.5 mM KCl, 1 mM CaCl2, 1 mM MgSO4, 0.15 mM KH2PO4, 0.05 mM NH2PO4, and 0.7 mM NaHCO3) and staged according to days post-fertilization (dpf) [2].
For chemical administration, zebrafish larvae at 5-7 dpf were exposed to ecabet sodium at concentrations of 0, 5, 10, 20, 40, and 80 μg/mL for 1 hour. After washing three times with embryo medium, neomycin was added at 125 μM for 1 hour. A negative control group received no additional chemicals. Larvae were then washed three times and anesthetized using triacaine (3-aminobenzoic acid 0.4 g/ethyl ester; 100 mL; pH 7 adjusted with Tris buffer) for 5 minutes. Larvae were mounted on a depression slide using methylcellulose for fluorescence microscopy assessment. Hair cells in four neuromasts (supraorbital SO1 and SO2, otic O1, and occipital OC1) on one side of each fish were counted under a fluorescence microscope at 40× magnification (n = 10 per group) [2].
For TUNEL assay, zebrafish larvae were exposed to 40 μg/mL ecabet sodium for 1 hour followed by 125 μM neomycin for 1 hour, then washed with phosphate buffer solution and fixed with 4% paraformaldehyde. They were incubated with 50 μL TUNEL reagent (TdT and fluorescein-dUTP) at 37°C for 60 minutes in a humid chamber. Apoptotic hair cells within the four neuromasts were assessed under a fluorescence microscope. TUNEL intensity was scored as: score 2 (almost all hair cells TUNEL-negative), score 1 (some TUNEL-positive), score 0 (many TUNEL-positive). Mean scores were compared (n = 15 per group) [2].
For ROS measurement, wild-type zebrafish larvae at 6 dpf were treated with 40 μg/mL ecabet sodium for 1 hour followed by 125 μM neomycin for 1 hour. After washing twice with PBS, larvae were transferred to 96-well plates and treated with DCFH-DA solution (20 μg/mL), then incubated for 1 hour in the dark at 28.5°C. Fluorescence intensity was quantified using a fluorescent microplate reader (n = 40) [2].
For DASPEI assay, wild-type zebrafish larvae were exposed to 40 μg/mL ecabet sodium for 1 hour followed by 125 μM neomycin for 1 hour. After anesthesia with triacaine, larvae were incubated in embryo medium containing 0.005% DASPEI for 15 minutes. DASPEI intensity of four neuromasts was assessed under a fluorescence microscope and graded as: score 2 (normal staining), score 1 (reduced staining), score 0 (absent staining). Mean scores were compared (n = 10 per group) [2].
For scanning electron microscopy, zebrafish were pre-fixed by immersion in 2% glutaraldehyde and 0.1 M phosphate buffer, then post-fixed for 2 hours in 1% osmotic acid dissolved in PBS. Larvae were then maintained in a graded series of ethanol and t-butyl alcohol, dried in a freeze dryer, and observed under a field emission SEM after platinum coating with an ion coater [2].

Ecabet sodium itself at a concentration of 80 μg/mL was toxic to hair cells compared to the negative control (p < 0.01). However, the 40 μg/mL concentration of ecabet sodium did not show toxicity to hair cells compared to the negative control (p = 0.21) [2].

[1]. Efficacy and safety of ecabet sodium as an adjuvant therapy for Helicobacter pylori eradication: a systematic review and meta-analysis. Helicobacter. 2014 Oct;19(5):372-81.

[2]. Ecabet sodium alleviates neomycin-induced hair cell damage. Free Radic Biol Med. 2015 Dec;89:1176-83.

[3]. Ecabet sodium inhibits Helicobacter pylori lipopolysaccharide-induced activation of NADPH oxidase 1 or apoptosis of guinea pig gastric mucosal cells. Am J Physiol Gastrointest Liver Physiol.

Ecabet sodium is a dehydroabietic acid derivative purified from pine resin. It is currently administered for the treatment of gastritis and gastric ulcer. The mechanisms include anti-bacterial activity, enhanced mucosal protection, and anti-peptic activity. Studies have shown that ecabet sodium inhibits reactive oxygen species (ROS) production. This study demonstrated that ecabet sodium protects against neomycin-induced hair cell loss in zebrafish, possibly by reducing apoptosis, mitochondrial damage, and ROS generation. To the authors' knowledge, this was the first study investigating the protective effects of ecabet sodium against neomycin-induced ototoxicity in zebrafish [2].

C20H26NA2O5S

424.46

402.147

C, 56.59; H, 6.17; Na, 10.83; O, 18.85; S, 7.55

86408-72-2

Ecabet;33159-27-2

23663982

Solid powder

1.241 g/cm3

1.562

4.889

1

5

3

27

672

3

C[C@]12CCC[C@@](C)(C(=O)O)[C@@H]1CCC1=CC(=C(C=C21)S(O)(=O)=O)C(C)C.[Na]

NQQLQPVASUVBKM-MUMAJMNHSA-L

InChI=1S/C20H28O5S.2Na/c1-12(2)14-10-13-6-7-17-19(3,8-5-9-20(17,4)18(21)22)15(13)11-16(14)26(23,24)25;;/h10-12,17H,5-9H2,1-4H3,(H,21,22)(H,23,24,25);;/q;2*+1/p-2/t17-,19-,20-;;/m1../s1

disodium;(1R,4aS,10aR)-1,4a-dimethyl-7-propan-2-yl-6-sulfonato-2,3,4,9,10,10a-hexahydrophenanthrene-1-carboxylate

Ecabet Sodium; TA-2711; TA-2711E; TA-2711; TA 2711

2934.99.9001

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, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 80~100 mg/mL (198.8~248.5 mM)
Ethanol: ~3 mg/mL (~7.5 mM) Water: ~16 mg/mL (~39.8 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.21 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (6.21 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.21 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)