a substrate of alcohol dehydrogenase class III isoenzyme
The primary target of GSNO is the nitric oxide (NO) signaling pathway. GSNO acts as a NO donor, releasing NO which then activates soluble guanylate cyclase (sGC), leading to the production of cyclic GMP (cGMP). cGMP mediates various physiological effects, including vasodilation, inhibition of platelet aggregation, and smooth muscle relaxation. GSNO also mediates protein S-nitrosylation, a post-translational modification that regulates protein function.

Nitrosoglutathione (GSNO; 250 μM) inhibited 90% of the reaction to 0.1 μM 5-HT and 40% of the response to 1.0 μM 5-HT in rings treated with LY-83583, suggesting that GSNO's effect is not dependent on guanylyl cyclase activity [5].
In vitro, GSNO has been shown to release NO in a controlled manner. It induces vasodilation in isolated blood vessels and inhibits platelet aggregation. GSNO also mediates S-nitrosylation of various proteins, affecting their function. The compound's effects on NO signaling have been confirmed in various in vitro models.

In PE-induced rats, nitrosoglutathione (GSNO, 8 mg/kg) can dramatically lower mean arterial pressure, diastolic blood pressure, and systolic blood pressure between days 14 and 20 [3]. Nitrosoglutathione (GSNO, 0.2 and 0.6 mg/kg) increased endothelial NOS expression while markedly reducing superoxide generation, NF-κB activation, iNOS induction, and 3-nitrotyrosine expression [4].
In vivo, GSNO is involved in the regulation of blood pressure, platelet function, and neurotransmission. It has been studied for its potential therapeutic applications in cardiovascular diseases, respiratory diseases, and conditions associated with NO deficiency. GSNO has been shown to have beneficial effects in animal models of hypertension, asthma, and ischemia-reperfusion injury.

An enzyme isolated from rat liver cytosol (native molecular mass 78. 3 kDa; polypeptide molecular mass 42.5 kDa) is capable of catalysing the NADH/NADPH-dependent degradation of S-nitrosoglutathione (GSNO). The activity utilizes 1 mol of coenzyme per mol of GSNO processed. The isolated enzyme has, as well, several characteristics that are unique to alcohol dehydrogenase (ADH) class III isoenzyme: it is capable of catalysing the NAD+-dependent oxidations of octanol (insensitive to inhibition by 4-methylpyrazole), methylcrotyl alcohol (stimulated by added pentanoate) and 12-hydroxydodecanoic acid, and also the NADH/NADPH-dependent reduction of octanal. Methanol and ethanol oxidation activity is minimal. The enzyme has formaldehyde dehydrogenase activity in that it is capable of catalysing the NAD+/NADP+-dependent oxidation of S-hydroxymethylglutathione. Treatment with the arginine-specific reagent phenylglyoxal prevents the pentanoate stimulation of methylcrotyl alcohol oxidation and markedly diminishes the enzymic activity towards octanol, 12-hydroxydodecanoic acid and S-hydroxymethylglutathione; the capacity to catalyse GSNO degradation is also checked. Additionally, limited peptide sequencing indicates 100% correspondence with known ADH class III isoenzyme sequences. Kinetic studies demonstrate that GSNO is an exceptionally active substrate for this enzyme. S-Nitroso-N-acetylpenicillamine and S-nitrosated human serum albumin are not substrates; the activity towards S-nitrosated glutathione mono- and di-ethyl esters is minimal. Product analysis suggests that glutathione sulphinamide is the major stable product of enzymic GSNO processing, with minor yields of GSSG and NH3; GSH, hydroxylamine, nitrite, nitrate and nitric oxide accumulations are minimal. Inclusion of GSH in the reaction mix decreases the yield of the supposed glutathione sulphinamide in favor of GSSG and hydroxylamine[2].
In vitro enzyme or receptor binding (non-cell) assays for GSNO involve measuring its NO-releasing activity. NO release can be measured using a chemiluminescence-based assay or by using a NO-sensitive electrode. The compound's ability to activate soluble guanylate cyclase (sGC) can be measured by assessing cGMP production in cell-free systems.

S-Nitrosation was induced in rat isolated middle cerebral arteries by pretreatment with the NO donors, S-nitrosoglutathione (GSNO) or sodium nitroprusside (SNP). Agonist-dependent activation of AT1 receptors was evaluated by obtaining concentration-response curves to AngII. Ligand-independent activation of AT1 receptors was evaluated by calculating MT (active vs. passive diameter) at pressures ranging from 20 to 200 mmHg in the presence or not of a selective AT1 receptor inverse agonist. Key results: GSNO or SNP completely abolished the AngII-dependent AT1 receptor-mediated vasoconstriction of cerebral arteries. GSNO had no impact on responses to other vasoconstrictors sharing (phenylephrine, U46619) or not (5-HT) the same signalling pathway. MT was reduced by GSNO, and the addition of losartan did not further decrease MT, suggesting that GSNO blocks AT1 receptor-dependent MT. Ascorbate (which reduces S-nitrosated compounds) restored the response to AngII but not the soluble GC inhibitor ODQ, suggesting that these effects are mediated by S-nitrosation rather than by S-nitrosylation.[1]
In vitro cell-based assays for GSNO are performed using various cell types, including endothelial cells, smooth muscle cells, and platelets. Cells are treated with the compound, and NO production, cGMP levels, and S-nitrosylation of proteins are measured. The compound's effects on cell function, such as vasodilation, platelet aggregation, and cell proliferation, are assessed.

Animal/Disease Models: Male Lewis rat [4].
Doses: 0.2 and 0.6 mg/kg.
Route of Administration: Slowly intravenously (iv) (iv)(iv) into each rat via the opposite femoral vein.
Experimental Results: Animals treated with 0.2 mg GSNO per kilogram before reperfusion had moderate survival (40.2 ± 4.9%). Although 0.6 mg/kg GSNO demonstrated a better rescue effect than 150 mg/kg NAC, there was no significant difference between groups.

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On day 14 of gestation, female Sprague-Dawley rats were separated into five groups and treated intravenously for 7 days as follows: (i) 0.3 mL 0.9% saline (control, n = 11); (ii) 50 mg/kg Body Weight (BW) N-nitro-L-arginine methyl ester (L-NAME) in 0.3 mL saline (n = 10); (iii) 50 mg/kg BW L-NAME and 8 mg/kg BW GSNO in 0.15 mL saline (n = 6); (iv) 50 mg/kg BW L-NAME in 0.15 mL saline and 8 mg/kg BW SNAP in 0.15 mL DMSO (n = 9); and (v) 0.15 mL DMSO and 0.15 mL saline (SNAP control, n = 7). Blood pressures were measured on day 14 through day 20, a 4-h urine sample was taken on day 20, and animals were sacrificed on day 21. Pups were counted and weighed individually. Results: SNAP and GSNO significantly decreased systolic, diastolic, and mean arterial pressures in PE-induced rats from day 14 through day 20 (P < 0.05). Pup weights in SNAP and GSNO groups were higher than in L-NAME group but lower than in controls (P ≤ 0.001). SNAP and GSNO partially reversed growth retardation.[3]

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Thirty minutes before flap reperfusion, normal saline, N-acetylcysteine (75 and 150 mg/kg), or GSNO (0.2 and 0.6 mg/kg) was randomly injected into 10 rats. Superoxide, nuclear factor-kappa B (NF-kappa B) activation, NO synthase (NOS) isoforms, and 3-nitrotyrosine expression in the pedicle vessels as well as survival areas of the flaps were evaluated. Results: I/R injury induced superoxide production, NF-kappa B activation, and inducible NOS (iNOS) expression in the pedicle vessels. GSNO significantly inhibited superoxide production and suppressed NF-kappa B activation, iNOS induction, and 3-nitrotyrosine expression, but up-regulated endothelial NOS expression in the flap vessels. Optimal doses of both GSNO (0.6 mg/kg) and N-acetylcysteine (150 mg/kg) effectively promoted flap survival area (p < 0.001), although there was no significant difference between both groups.[4]
In vivo animal experiments for GSNO are conducted using rodent models of cardiovascular and respiratory diseases. The compound is administered intravenously, intraperitoneally, or by inhalation. Blood pressure, platelet function, and airway resistance are measured. The compound's effects on disease pathology are assessed in tissue samples.

Pharmacokinetic (PK) properties of GSNO indicate that it is a labile compound with a short half-life. The compound has a molecular weight of 336.32 and a molecular formula of C10H16N4O7S. It is soluble in water and DMSO. GSNO is rapidly metabolized, and its stability is pH-dependent. The compound is typically stored at -20°C under inert atmosphere.

Toxicology (toxicology) data for GSNO indicate that it is generally well-tolerated at therapeutic doses. As an NO donor, it can cause hypotension and vasodilation at high doses. The compound may also have pro-oxidant effects due to the formation of reactive nitrogen species. Long-term safety data are being evaluated in clinical studies.

[1]. S-nitrosoglutathione inhibits cerebrovascular angiotensin II-dependent and -independent AT 1 receptor responses: A possible role of S-nitrosation. Br J Pharmacol. 2019 Jun;176(12):2049-2062.
[2]. S-Nitrosoglutathione is a substrate for rat alcohol dehydrogenase class III isoenzyme. Biochem J. 1998 Apr 15;331 ( Pt 2)(Pt 2):659-68.
[3]. The effects of S-nitrosoglutathione and S-nitroso-N-acetyl-D, L-penicillamine in a rat model of pre-eclampsia. J Nat Sci Biol Med. 2013 Jul;4(2):330-5.
[4]. Nitrosoglutathione promotes flap survival via suppression of reperfusion injury-induced superoxide and inducible nitric oxide synthase induction. J Trauma. 2004 Nov;57(5):1025-31.
[5]. Pulmonary vasoconstriction by serotonin is inhibited by S-nitrosoglutathione. Am J Physiol Lung Cell Mol Physiol. 2002 May;282(5):L1057-65.

S-nitrosoglutathione is a glutathione derivative in which the hydrogen atom bonded to the sulfur atom is replaced by a nitroso group. It has functions including inhibiting platelet aggregation, dilating bronchi, acting as a nitric oxide donor, and serving as a signaling molecule. It is a glutathione derivative and also a nitrosothio compound. It is the conjugate acid of S-nitrosoglutathione (2-). It is a sulfur-containing alkyl nitrite, belonging to the nitric oxide donor group.
Other information: GSNO is an endogenous NO donor and S-nitrosothiol. It is also known as S-Nitrosoglutathione. The compound is used in research to study NO signaling and as a potential therapeutic agent for cardiovascular and respiratory diseases. Its CAS number is 57564-91-7.

C10H16N4O7S

336.31

336.073

C, 35.71; H, 4.80; N, 16.66; O, 33.30; S, 9.53

57564-91-7

104858

Pink to red solid

1.7±0.1 g/cm3

>170ºC (dec.)

1.656

-0.26

5

10

10

22

445

2

O=NSC[C@@H](C(NCC(O)=O)=O)NC(CC[C@H](N)C(O)=O)=O

HYHSBSXUHZOYLX-WDSKDSINSA-N

InChI=1S/C10H16N4O7S/c11-5(10(19)20)1-2-7(15)13-6(4-22-14-21)9(18)12-3-8(16)17/h5-6H,1-4,11H2,(H,12,18)(H,13,15)(H,16,17)(H,19,20)/t5-,6-/m0/s1

N5-((R)-1-((carboxymethyl)amino)-3-(nitrosothio)-1-oxopropan-2-yl)-L-glutamine

S-Nitrosoglutathione; SNOG; RVC-588; S-Nitroso-L-glutathione; Nitrosoglutathione; Glutathione thionitrite; (S)-2-Amino-5-(((R)-1-((carboxymethyl)amino)-3-(nitrosothio)-1-oxopropan-2-yl)amino)-5-oxopentanoic acid; N-(N-L-gamma-Glutamyl-S-nitroso-L-cysteinyl)glycine; GSNO

2934.99.9001

Powder      -80°C    3 years

                     -20°C     1 year

Note: This product is not stable under room temperature, please store the powder form in -20 °C or -80 °C and freshly prepare solutions (from powder form) immediately before performing experiments for optimal results.

On dry ice or blue ice (Note: This product is not stable under room temperature, please store it under -20 °C or -80 °C immediately after receiving it)

H2O : ~25 mg/mL (~74.33 mM)

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.)