Glutathione trisulfide TFA (GSSSG TFA) is an orally active, blood-brain barrier permeable neuroprotective agent. Glutathione trisulfide TFA inactivates intracellular tyrosinase, regulates the expression of Cars2, Cbs, MITF and TYR, inhibits α-MSH -induced melanogenesis, and restores intracellular persulfide levels reduced by α-MSH. Glutathione trisulfide TFA scavenges free radicals, quenches ROS, reduces Paclitaxel -induced superoxide production, upregulates the expression of antioxidant protein genes, and inhibits oxidative stress-induced cell death. Glutathione trisulfide TFA promotes ERK1/2 activation, prevents NF-κB p65 activation, inhibits TAK1 phosphorylation, reduces pro-inflammatory cytokine expression, and blocks microglial activation. Glutathione trisulfide TFA can be used in research related to dry age-related macular degeneration, inflammation-associated eye diseases, Alzheimer's disease, Parkinson's disease, etc. [1] [2] [3] [4] [5]
In Vivo, Glutathione trisulfide (15-60 nmol/eye; intravitreal; single co-administration with LPS) TFA (GSSSG TFA) exerts dose-dependent anti-inflammatory activity in LPS-induced rat retinal inflammation, with 60 nmol/eye significantly reducing proinflammatory cytokine mRNA upregulation and microglial accumulation[2]. Glutathione trisulfide (200 mg/kg; i.p.; daily; 4 days) TFA prevents post-ischemic spinal cord injury-induced paraplegia in mice, while maintaining a 100% survival rate[3]. Glutathione trisulfide (50 mg/kg/day; oral gavage; daily; 28 days) TFA ameliorates Paclitaxel-induced mechanical allodynia in male C57BL/6J mice by preserving unmyelinated axon integrity, reducing mitochondrial swelling, and upregulating antioxidant signaling in peripheral sensory neurons[5].
In Vitro, Glutathione trisulfide (25-100 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA (GSSSG TFA) is non-cytotoxic to B16-F0 cells at 25 μM and 50 μM following 30 min pretreatment and 24 h α-MSH incubation, while 100 μM causes a modest non-significant viability reduction[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA potently suppresses α-MSH-induced melanin production in B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA dose-dependently downregulates Mitf and TYR gene expression in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly upregulates Cars2 and Cbs gene expression in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly restores α-MSH-reduced intracellular persulphide levels in B16-F0 cells[1]. Glutathione trisulfide (50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly inactivates intracellular tyrosinase activity in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (0.1-100 μM; 24 h) TFA protects SH-SY5Y cells from MPP+-induced cell death[3]. Glutathione trisulfide (0.1-2 mM; 30 min) TFA exhibits dose-dependent antioxidant radical-scavenging activity in a cell-free DPPH assay, with up to 65% scavenging activity at 2 mM[1]. Glutathione trisulfide (10-30 μM; 30 minutes) TFA attenuates Paclitaxel-induced superoxide production in cultured murine primary cortical neurons[5]. Glutathione trisulfide (25-200 µM; 6 h) TFA is non-toxic to mouse primary Müller cells and BV-2 microglial cells at concentrations up to 100 µM, while 200 µM reduces Müller cell viability[2]. Glutathione trisulfide (25-200 µM; 1 h pre-incubation, 6 h or 24 h LPS stimulation) TFA dose-dependently inhibits LPS-induced upregulation of IL-6 and Ccl2 mRNA expression and protein secretion in mouse primary Müller cells, with significant effects at concentrations of 100 and 200 µM[2]. Glutathione trisulfide (200 µM; 1 h pre-incubation, 30 or 60 min LPS stimulation) TFA inhibits LPS-induced phosphorylation of TAK1 (Ser412) in mouse BV-2 microglial cells at 30 and 60 minutes post-LPS stimulation[2]. Glutathione trisulfide (25-200 µM; 1 h pre-incubation, 6 h or 24 h LPS stimulation) TFA dose-dependently inhibits LPS-induced upregulation of TNF-α, Ccl2, IL-6, and IL-1β mRNA expression, as well as TNF-α, Ccl2, and IL-6 protein secretion in mouse BV-2 microglial cells, with significant effects starting at 25 µM for some targets[2]. Glutathione trisulfide (500 nM; 1 hour) TFA prevents Paclitaxel -induced axonal degeneration and mitochondrial fragmentation in cultured murine primary DRG neurons by restoring axonal elongation and preserving axonal mitochondrial integrity[5]. Glutathione trisulfide (10 μM; 24 hours) TFA does not alter the anti-tumor cytotoxic effect of 2 μM Paclitaxel on human breast cancer MDA-MB-231 cells[5].
References:
[1]. Uchida Y, et al. Glutathione Trisulphide Improves Skin Brightness with Anti‑Melanogenesis Effects and Maintains Intracellular Persulphide Levels. Cosmetics. 2025 Oct 21;12(5):234.
[2]. Tawarayama H, et al. Glutathione trisulfide prevents lipopolysaccharide-induced retinal inflammation via inhibition of proinflammatory cytokine production in glial cells. Scientific reports. 2023 Jul 17;13(1):11513.
[3]. Ichinose F, et al. Glutathione trisulfide (GSSSG) in neuroprotection. WO2021231476 A1. 2021 Nov 18.
[4]. Brown EM, et al. Stabilities of Three Key Biological Trisulfides with Implications for Their Roles in the Release of Hydrogen Sulfide and Bioaccumulation of Sulfane Sulfur. ACS omega. 2022 Apr 05;7(13):11440-11451.
[5]. Ezaka M, et al. Oral Administration of Glutathione Trisulfide Increases Reactive Sulfur Levels in Dorsal Root Ganglion and Ameliorates Paclitaxel-Induced Peripheral Neuropathy in Mice. Antioxidants (Basel, Switzerland). 2022 Oct 27;11(11):2122.
In Vivo, Glutathione trisulfide (15-60 nmol/eye; intravitreal; single co-administration with LPS) TFA (GSSSG TFA) exerts dose-dependent anti-inflammatory activity in LPS-induced rat retinal inflammation, with 60 nmol/eye significantly reducing proinflammatory cytokine mRNA upregulation and microglial accumulation[2]. Glutathione trisulfide (200 mg/kg; i.p.; daily; 4 days) TFA prevents post-ischemic spinal cord injury-induced paraplegia in mice, while maintaining a 100% survival rate[3]. Glutathione trisulfide (50 mg/kg/day; oral gavage; daily; 28 days) TFA ameliorates Paclitaxel-induced mechanical allodynia in male C57BL/6J mice by preserving unmyelinated axon integrity, reducing mitochondrial swelling, and upregulating antioxidant signaling in peripheral sensory neurons[5].
In Vitro, Glutathione trisulfide (25-100 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA (GSSSG TFA) is non-cytotoxic to B16-F0 cells at 25 μM and 50 μM following 30 min pretreatment and 24 h α-MSH incubation, while 100 μM causes a modest non-significant viability reduction[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA potently suppresses α-MSH-induced melanin production in B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA dose-dependently downregulates Mitf and TYR gene expression in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly upregulates Cars2 and Cbs gene expression in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (25-50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly restores α-MSH-reduced intracellular persulphide levels in B16-F0 cells[1]. Glutathione trisulfide (50 μM; 30 min pretreatment, 24 h α-MSH incubation) TFA significantly inactivates intracellular tyrosinase activity in α-MSH-induced B16-F0 cells[1]. Glutathione trisulfide (0.1-100 μM; 24 h) TFA protects SH-SY5Y cells from MPP+-induced cell death[3]. Glutathione trisulfide (0.1-2 mM; 30 min) TFA exhibits dose-dependent antioxidant radical-scavenging activity in a cell-free DPPH assay, with up to 65% scavenging activity at 2 mM[1]. Glutathione trisulfide (10-30 μM; 30 minutes) TFA attenuates Paclitaxel-induced superoxide production in cultured murine primary cortical neurons[5]. Glutathione trisulfide (25-200 µM; 6 h) TFA is non-toxic to mouse primary Müller cells and BV-2 microglial cells at concentrations up to 100 µM, while 200 µM reduces Müller cell viability[2]. Glutathione trisulfide (25-200 µM; 1 h pre-incubation, 6 h or 24 h LPS stimulation) TFA dose-dependently inhibits LPS-induced upregulation of IL-6 and Ccl2 mRNA expression and protein secretion in mouse primary Müller cells, with significant effects at concentrations of 100 and 200 µM[2]. Glutathione trisulfide (200 µM; 1 h pre-incubation, 30 or 60 min LPS stimulation) TFA inhibits LPS-induced phosphorylation of TAK1 (Ser412) in mouse BV-2 microglial cells at 30 and 60 minutes post-LPS stimulation[2]. Glutathione trisulfide (25-200 µM; 1 h pre-incubation, 6 h or 24 h LPS stimulation) TFA dose-dependently inhibits LPS-induced upregulation of TNF-α, Ccl2, IL-6, and IL-1β mRNA expression, as well as TNF-α, Ccl2, and IL-6 protein secretion in mouse BV-2 microglial cells, with significant effects starting at 25 µM for some targets[2]. Glutathione trisulfide (500 nM; 1 hour) TFA prevents Paclitaxel -induced axonal degeneration and mitochondrial fragmentation in cultured murine primary DRG neurons by restoring axonal elongation and preserving axonal mitochondrial integrity[5]. Glutathione trisulfide (10 μM; 24 hours) TFA does not alter the anti-tumor cytotoxic effect of 2 μM Paclitaxel on human breast cancer MDA-MB-231 cells[5].
References:
[1]. Uchida Y, et al. Glutathione Trisulphide Improves Skin Brightness with Anti‑Melanogenesis Effects and Maintains Intracellular Persulphide Levels. Cosmetics. 2025 Oct 21;12(5):234.
[2]. Tawarayama H, et al. Glutathione trisulfide prevents lipopolysaccharide-induced retinal inflammation via inhibition of proinflammatory cytokine production in glial cells. Scientific reports. 2023 Jul 17;13(1):11513.
[3]. Ichinose F, et al. Glutathione trisulfide (GSSSG) in neuroprotection. WO2021231476 A1. 2021 Nov 18.
[4]. Brown EM, et al. Stabilities of Three Key Biological Trisulfides with Implications for Their Roles in the Release of Hydrogen Sulfide and Bioaccumulation of Sulfane Sulfur. ACS omega. 2022 Apr 05;7(13):11440-11451.
[5]. Ezaka M, et al. Oral Administration of Glutathione Trisulfide Increases Reactive Sulfur Levels in Dorsal Root Ganglion and Ameliorates Paclitaxel-Induced Peripheral Neuropathy in Mice. Antioxidants (Basel, Switzerland). 2022 Oct 27;11(11):2122.