S3226 can be used as a selective inhibitor of NHE3 in mouse fibroblast L cells, opossum kidney cells and pig kidney brush border membrane vesicles[2]. S3226 can effectively inhibit rat NHE3 in rat fibroblasts transfected with rat NHE3, with an IC50 of 0.2 μM[3].

S3226 (4-30 μM; intratubular microperfusion; single dose per nephron; equilibration 5 minutes before measurement) inhibits proximal tubular fluid and Na+ reabsorption, with an IC50 of 4-5 μM and a maximum inhibition rate of approximately 30%, but does not affect loop of Henry reabsorption or tubuloglomerular feedback in anesthetized rats [2]. S3226 (20 mg/kg; intravenous injection; within 30 minutes) improves glomerular filtration rate, renal electrolyte handling and renal histology, and increases the survival rate of ischemia-induced acute renal failure rats to 100% [3]. S3226 (20 mg/kg; intravenous injection; within 30 minutes; two doses) significantly reduces intracellular renal pH by 0.15 units during reperfusion in rats with ischemic acute renal failure after unilateral nephrectomy [3].

Animal/Disease Models:Munich-Vista-Fromt mice (adult males, 270-350 g, anesthetized, superficial nephrons) [2]
Doses: 4-5 µM; 10 µM; 30 µM
Route of Administration: Intratubular microperfusion; single dose per nephron; equilibration for 5 minutes before measurement
Experimental Results: Dose-dependent inhibition of reabsorption of fluid, Na+, and Cl- in the proximal tubules, IC50 of 4-5 µM. The maximum inhibition rate of fluid and Na+ reabsorption was approximately 30%, and the maximum inhibition rate of Cl- reabsorption was approximately 10%. At concentrations of 10 µM or 30 µM, the reduction in Na+ reabsorption was significantly greater than the reduction in Cl- reabsorption. At concentrations of 10 µM and 30 µM, there was no effect on the reabsorption of fluid, Na+, or K+ in the loop of Henle. At a concentration of 30 µM, the tubuloglomerular feedback response was not altered, including the amplitude of the cessation pressure, the slope of the curve, or the shift in the operating point.

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Animal/Disease Models:Wistar (male, average weight 285 g, bilateral renal artery occlusion for 40 minutes) [3]
Doses: 20 mg/kg
Route of Administration: Intravenous injection; within 30 minutes; single dose (before occlusion or after reperfusion)
Experimental Results: Compared with the control group of 0.30 mL/min/kg, creatinine clearance increased to 0.90 mL/min/kg (before occlusion) and 0.83 mL/min/kg (after reperfusion) on day 1 after ischemia. On day 1, sodium excretion fraction decreased from 8.17% in the control group to 1.42% in the pre-occlusion treatment group and 1.88% in the post-reperfusion treatment group. On day 1, urinary sodium excretion decreased to 0.31 mmol/kg in the pre-occlusion treatment group and 0.31 mmol/kg in the post-reperfusion treatment group, compared to 0.98 mmol/kg in the control group. Compared to the control group, the increase in plasma creatinine on days 1, 2, and 3 after infusion was reduced. Plasma potassium concentration decreased on days 2 and 3 compared to the control group. The fractional potassium excretion on day 1 decreased compared to the control group. Kidney tissue collected on day 7 showed significantly reduced tubular necrosis, dilation, protein casts, and cellular infiltration. The survival rate reached 100% within 7 days, compared to 75% in the control group.

[1]. Na+ / H+ exchanger-3 is involved in mouse blastocyst formation. J Exp Zool A Comp Exp Biol. 2004;301(9):767-775.

[2]. Role of Na+/H+ exchanger NHE3 in nephron function: micropuncture studies with S3226, an inhibitor of NHE3[J]. American Journal of Physiology-Renal Physiology, 2000, 278(3): F375-F379.

[3]. S3226, a novel NHE3 inhibitor, attenuates ischemia-induced acute renal failure in rats. Kidney Int. 2001;60(6):2283-2289.

C17H24CL2N6O2

415.32

215183-03-2

White to off-white solid

[H]Cl.[H]Cl.C/C(C(NC(N)=N)=O)=C\C(C=CC(C)=C1)=C1/C=C(C)/C(NC(N)=N)=O

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: 请将本产品存放在密封且受保护的环境中（例如氮气下），避免暴露在潮湿环境中。

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

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

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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