| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
The compound modulated the expression and activity of several proteins involved in metastasis: matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), urokinase plasminogen activator (uPA), tissue inhibitor of metalloproteinase-1 (TIMP-1), tissue inhibitor of metalloproteinase-2 (TIMP-2), non-metastatic protein 23 homologue 1 (nm23-H1), plasminogen activator inhibitor-1 (PAI-1), and proliferating cell nuclear antigen (PCNA).
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| ln Vivo |
vit CK3 significantly and dose-dependently inhibited tumor growth and lung metastasis in LLC-bearing mice. Low-dose vit CK3 (100 mg vit C/kg + 1 mg vit K3/kg) and high-dose vit CK3 (1000 mg vit C/kg + 10 mg vit K3/kg) reduced tumor volumes starting from 2 weeks after tumor cell implantation. Tumor weights were significantly reduced by both doses (low-dose: 4.56 ± 1.33 g; high-dose: 4.32 ± 0.76 g) compared to tumor control (8.90 ± 2.48 g). vit CK3 restored body weight of tumor-bearing mice to the level of tumor-free mice. vit CK3 significantly decreased plasma activities of MMP-2, MMP-9, and uPA. In lung tissues, vit CK3 increased protein expression of TIMP-1, TIMP-2, nm23-H1, and PAI-1; reduced protein expression of MMP-2 and MMP-9; and inhibited PCNA expression. High-dose vit CK3 significantly reduced relative spleen and lung weights compared to tumor control. Lung metastasis incidence: tumor control 10/10 (100%), low-dose vit CK3 9/10 (90%), high-dose vit CK3 8/10 (80%). Tumor area in lung: tumor control 28.3 ± 2.2%, low-dose vit CK3 14.6 ± 4.9%, high-dose vit CK3 8.7 ± 2.3%. Number of lung metastases: tumor control 13.4 ± 4.0, low-dose vit CK3 7.1 ± 2.3, high-dose vit CK3 3.8 ± 2.8. [1]
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| Enzyme Assay |
MMP-2, MMP-9, and u-PA activities in plasma were measured by gelatin and casein plasminogen zymography protease assays. Plasma samples were diluted 1:20 with PBS immediately before assay. After electrophoresis, gels were washed with 2.5% Triton X-100 and then incubated in reaction buffer (40 mM Tris-HCl, pH 8.0; 10 mM CaCl2; 0.01% NaN3) for 12-15 h at 37°C. Gels were stained with Coomassie brilliant blue R-250 for 30 min and then destained in 10% acetic acid (v/v) and 50% methanol (v/v). Relative MMP and u-PA activities were quantified by software. [1]
Protein expression of TIMP-1, TIMP-2, MMP-2, MMP-9, nm23-H1, and PAI-1 in lung tissues was measured by Western blotting. Lung tissues were homogenized in cold RIPA buffer containing protease inhibitors and centrifuged (10,000 × g; 5 min) at 4°C. Supernatants were frozen at -80°C. Protein (80 μg) from supernatant was resolved by SDS-PAGE and transferred onto a PVDF membrane. The membrane was blocked with 5% nonfat milk in TBS buffer (20 mmol/l Tris-HCl, 150 mmol/l NaCl, pH 7.4) for 1 h, then incubated with monoclonal antibody followed by horseradish peroxidase-conjugated anti-mouse IgG, and visualized using an ECL chemiluminescent detection kit. [1] |
| Cell Assay |
The mouse Lewis lung carcinoma (LLC) cell line was cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% (v:v) fetal bovine serum (FBS), 0.37% (w/v) NaHCO3, penicillin (100 kU/l), and streptomycin (100 kU/l) in a humidified incubator under 5% CO2 and 95% air at 37°C. Approximately 2 × 10^6 LLC cells (0.1 mL/mouse) were injected subcutaneously into the right flank of C57BL/6 male mice for tumor implantation. [1]
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| Animal Protocol |
C57BL/6 male mice (5-week-old; 25-30 g) were housed individually in cages with controlled temperature (25 ± 2°C), humidity (65 ± 5%) with 12-h light/dark cycles. Approximately 2 × 10^6 LLC cells (0.1 mL/mouse) were injected subcutaneously into the right flank. Nine days after implantation, mice were randomly divided into 5 groups (N=8-10 per group): blank control, tumor control, low-dose vit CK3 (100 mg vit C/kg + 1 mg vit K3/kg, i.p.), high-dose vit CK3 (1000 mg vit C/kg + 10 mg vit K3/kg, i.p.), and positive control cisplatin (6 mg/kg, i.p.). Mice were treated via intraperitoneal injection twice a week for additional 28 days. During accommodation and experimental periods, mice were supplied a standard rodent diet (containing 59.8% carbohydrate, 23.4% protein, 4.5% crude fat without vit C and vit K3) and water ad libitum. Tumor growth was measured by caliper twice a week for 28 days. Tumor volume formula: Length × Width × Width/2. At the end, mice were euthanized with CO2 asphyxiation, primary tumors isolated and weighed. Blood samples collected from retroorbital plexus into K3EDTA tubes, centrifuged (4000 × g; 10 min) to obtain plasma. All lung lobes were microscopically examined for metastasis. [1]
For immunohistochemical assay of PCNA, lung tissues were formalin-fixed, embedded in paraffin, 2-μm sectioned, and subjected to H&E or IHC staining. IHC staining for PCNA was performed using the streptavidin-peroxidase technique. Positive rates (%) of PCNA stained cells were quantified by software. [1] |
| Toxicity/Toxicokinetics |
vit CK3 supplementation produced no overt toxicity as evidenced by lack of decreased body weights during the entire experimental period compared to tumor-free mice. Final body weights: low-dose vit CK3 24.6 ± 4.2 g, high-dose vit CK3 24.5 ± 2.9 g, not significantly different from blank control (25.2 ± 1.2 g) or tumor control (23.5 ± 4.8 g). In contrast, cisplatin-treated mice showed significantly lower final body weight (14.6 ± 1.6 g) compared to blank control, indicating marked side effects. High-dose vit CK3 significantly lowered relative spleen weight (0.83 ± 0.12%) and lung weight (1.07 ± 0.10%) compared to tumor control (spleen: 1.25 ± 0.33%; lung: 1.35 ± 0.30%). [1]
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| References | |
| Additional Infomation |
vit CK3 has been shown to inhibit tumor development and lung metastasis in several models of tumor-bearing mice. The antimetastatic effects may involve inhibition of MMP activities and protein expression, promotion of TIMPs (TIMP-1, TIMP-2) protein expression, increased protein expression of nm23-H1 and PAI-1, and inhibition of cell proliferation (decreased PCNA expression). The high dose used in mice (286 mg vit C/kg/day and 2.86 mg vit K3/kg/day) translates to a human equivalent dose (HED) of 1.6 g vit C and 16 mg vit K3 per 60-kg person per day. The upper intake level (UL) of vit C is 2 g/day for a healthy person; the calculated vit C dose (1.6 g/day) is safe for human supplementation, but safety of the calculated vit K3 dose (16 mg/day) is unclear. vit CK3 was administered intraperitoneally twice a week without causing obvious side effects. [1]
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| Molecular Formula |
C17H18NA2O11S
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|---|---|
| Molecular Weight |
476.362766742706
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| Exact Mass |
476.036
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| CAS # |
1085703-32-7
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| PubChem CID |
134128284
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
31
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| Complexity |
693
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CC1(CC(=O)C2=CC=CC=C2C1=O)S(=O)(=O)O.C([C@@H]([C@@H]1C(=C(C(=O)O1)O)O)O)O.[Na].[Na]
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| InChi Key |
RXNIARIYWPFYPK-NFAFUWKUSA-N
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| InChi Code |
InChI=1S/C11H10O5S.C6H8O6.2Na/c1-11(17(14,15)16)6-9(12)7-4-2-3-5-8(7)10(11)13;7-1-2(8)5-3(9)4(10)6(11)12-5;;/h2-5H,6H2,1H3,(H,14,15,16);2,5,7-10H,1H2;;/t;2-,5+;;/m.0../s1
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| HS Tariff Code |
2934.99.9001
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| Storage |
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. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~10 mg/mL (~21.08 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.11 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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. Solubility in Formulation 2: ≥ 1 mg/mL (2.11 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 10.0 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. View More
Solubility in Formulation 3: ≥ 1 mg/mL (2.11 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0993 mL | 10.4963 mL | 20.9925 mL | |
| 5 mM | 0.4199 mL | 2.0993 mL | 4.1985 mL | |
| 10 mM | 0.2099 mL | 1.0496 mL | 2.0993 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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