Mitochondrial-targeting antioxidant

Visomitin (SkQ1) administration of tumor-infiltrating leukocytes does not affect their cytotoxicity against Panc02 cells. At 500 nM concentration, visomitin significantly inhibits the growth of human PDAC cells while having no effect on the viability of the cell lines[1].

In reference to systemic angiogenic factors, KC is shown to be lower in the group receiving continuous therapy with visomitin (SkQ1) in the serum of mice having pancreatic ductal adenocarcinoma (PDAC). Visomitin treatment raises the quantity of VEGF molecules in the mice. Prolactin and MIP1a levels are lowered following the follow-up treatment or in all Visomitin treatment groups, respectively. Furthermore, IL-6 and IL-13 levels are higher in the groups that received visomitin treatment. The pretreatment setting results in a decrease in TGF-b levels. Conversely, every Visomitin treatment plan reduces the percentage of NKT cells. The PDAC-bearing mice's median survival has increased with Visomitin treatment, however the change is not statistically significant[1].

Panc02 cells are treated 48 h with different concentrations of Visomitin (SkQ1). Cell viability after Visomitin treatment is measured with an EZ4U Kit as described by the manufacturers. Briefly, 20,000 cells per well are seeded in 96-wellplates and let grow overnight. Afterwards, cells are treated without the medium exchange. A substrate compound from the kit is added and the cells are further incubated for 5 hr at 37°C to convert the yellow colored tetrazolium to its red formazan derivate by living cells. The absorbance is measured at 450 nm[1].

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Proliferation of cell lines was analyzed with a Bromodeoxyuridine (BrdU) Cell Proliferation Assay kit according manufacturer instructions. Briefly, 20,000 cells were seeded in 96-well plates and let grow overnight. After incubation with SKQ, the BrdU reagent was added, and the cells were incubated further for 12 hr at 37°C to allow the BrdU incorporation into proliferating cells. Afterwards, the cells were fixed, washed, and a detector antibody was added. The plates were incubated for 1 hrs at room temperature and washed. A goat anti-mouse IgG peroxidase conjugate from the kit was added, and the plates were incubated for 30 min at room temperature. After further washing, the cells were incubated for 30 min at room temperature in the dark with the 3,3′,5,5′-tetramethylbenzidine peroxidase substrate. The reaction was stopped by adding the acid stop solution from the kit. The absorbance was measured at 450 nm.[1]

SkQ1 treatment of mice[1]
Drinking water for C57BL/6 mice was supplemented with SkQ1 to yield a dose of 5 nmol SkQ1/kg body weight per day (on average, a mouse drank about 5 mL of water per day).12 The following experimental groups have been used: (1) control group (no SkQ1 in drinking water); (2) pretreatment group (animals treated with SkQ1 3 weeks before operation, no SkQ1 after Panc02 cell injection); (3) treatment group (animals received SkQ1 only after operation, without pretreatment); and (4) continuous treated group consisted of mice treated with SkQ1 both before and after tumor cell transplantation.

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For experiments on chronic pancreatitis, mice received cerulein (50 μg/kg/injection in saline) or saline (control) at five hourly injections of cerulein three times a week over a period of eight weeks. Antioxidative treatment with SkQ1 (10-(6′-plastoquinonyl)decyltriphenylphosphonium) was administered perorally with the drinking water at a dose of 5 nmol/kg body weight per day (on average, a mouse drank about 5 mL of water per day).[2]

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For experiments on both acute and chronic pancreatitis, mice were divided in three groups. Group A (acute pancreatitis (AP) n = 8; chronic pancreatitis (CP) n = 12) was treated with 5 nmol/kg SkQ1, group B (AP n = 8; CP n = 12) was the untreated control, and group C (AP n = 8; CP n = 7) was the sham group, which was injected intraperitoneally with 0.9% NaCl instead of cerulein and was therefore the negative control group without pancreatitis.[2]

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For experiments on acute pancreatitis, mice were pretreated with SkQ1 for 8 weeks prior to induction of pancreatitis. Mice designated for experiments on chronic pancreatitis received SkQ1 at the same concentration for 8 weeks in parallel with induction of pancreatitis.[2]

[1]. The novel mitochondria-targeted antioxidant SkQ1 modulates angiogenesis and inflammatory micromilieu in a murine orthotopic model of pancreatic cancer. Int J Cancer. 2016 Jul 1;139(1):130-9.

[2]. The Analgesic Effect of the Mitochondria-Targeted Antioxidant SkQ1 in Pancreatic Inflammation. Oxid Med Cell Longev. 2016;2016:4650489.

Our understanding in the last few years about reactive oxygen species (ROS) has changed from being harmful substances to crucial intra- and extracellular messengers as well as important regulators controlling a wide spectrum of signaling pathways, including those in cancer immunology. Therefore, these multiple essential roles of ROS and especially of mitochondria-derived ROS in malignant transformation and cancer progression make them a promising target for anticancer therapy. Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers in the world. A link between ROS, antioxidants and the PDAC development and progression has been recently established. Therefore, usage of specific highly efficient antioxidants could bring an option for treatment and/or prevention of PDAC. 10-(6'-plastoquinonyl) decyltriphenylphosphonium (SkQ1) is a new antioxidant with the highest mitochondrion membrane penetrating ability and potent antioxidant capability. In this work, we investigated an impact of SkQ1 on tumor angiogenesis, immune micromilieu, and oncological parameters in the orthotopic Panc02 murine model of PDAC. We showed that in this model SkQ1 treatment leads to the elevation of pro-angiogenic factors and to building of mainly an anti-inflammatory cytokine milieu. On the cellular level we showed an increase in a percentage of memory T cells and a decrease in frequency on natural killer T (NKT) cells. At the same time, SkQ1 was ineffective in the improvement of oncological parameters of PDAC-bearing mice. New studies are needed to clarify the absence of therapeutic and/or prophylactic benefits of the antioxidant.[1]

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Background. Chronic pancreatitis is one of the main risk factors for pancreatic cancer. In acute and chronic pancreatitis, oxidative stress is thought to play a key role. In this respect, the recently described mitochondria-targeted antioxidant SkQ1 effectively scavenges reactive oxygen species at nanomolar concentrations. Therefore, we aimed to characterize the influence of SkQ1 on tissue injury and pain in acute and chronic pancreatitis. Methods. Both acute and chronic pancreatitis were induced in C57BL/6 mice by intraperitoneal cerulein injections and treatment with SkQ1 was carried out by peroral applications. Hyperalgesia was assessed by behavioral observation and measurement of abdominal mechanical sensitivity. Blood serum and pancreatic tissue were harvested for analysis of lipase and histology. Results. SkQ1 did not influence pain, serological, or histological parameters of tissue injury in acute pancreatitis. In chronic pancreatitis, a highly significant reduction of pain-related behavior (p < 0.0001) was evident, but histological grading revealed increased tissue injury in SkQ1-treated animals (p = 0.03). Conclusion. After SkQ1 treatment, tissue injury is not ameliorated in acute pancreatitis and increased in chronic pancreatitis. However, we show an analgesic effect in chronic pancreatitis. Further studies will need to elucidate the risks and benefits of mitochondria-targeted antioxidants as an analgesic.[2]

C36H42BRO2P

617.61

616.21

C, 70.01; H, 6.85; Br, 12.94; O, 5.18; P, 5.02

934826-68-3

934826-68-3 (bromide);934960-96-0 (cation);1372443-45-2 (chloride);1372443-48-5 (sulfate); 714085-40-1 (iodide);

16679091

Yellow to brown solid powder

8.864

0

3

14

40

804

0

WYHFWTRUGAFNKW-UHFFFAOYSA-M

InChI=1S/C36H42O2P.BrH/c1-29-30(2)36(38)31(28-35(29)37)20-12-7-5-3-4-6-8-19-27-39(32-21-13-9-14-22-32,33-23-15-10-16-24-33)34-25-17-11-18-26-34/h9-11,13-18,21-26,28H,3-8,12,19-20,27H2,1-2H31H/q+1/p-1

(10-(4,5-dimethyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)triphenylphosphonium bromide

SKQ1; SKQ-1; SKQ 1; 934826-68-3; SKQ1 bromide; SKQ-1 bromide; SKQ1; 934826-68-3 (bromide); (10-(4,5-dimethyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)triphenylphosphonium bromide; 7B14500J3E; PDTP; Plastoquinonyl decyltriphenyl phosphonium bromide; Visomitin

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 (e.g. under nitrogen), 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 : ~100 mg/mL (~161.92 mM)
Ethanol : ~50 mg/mL (~80.96 mM)
H2O : ~3.33 mg/mL (~5.39 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.05 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 (4.05 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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 (4.05 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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 4: ≥ 2.5 mg/mL (4.05 mM) (saturation unknown) in 10% EtOH + 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 EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix well.
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 5: ≥ 2.5 mg/mL (4.05 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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.)