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Purity: ≥98%
NSC-95397 is a novel, potent, and selective CDC25B inhibitor which inhibits influenza A virus replication in dose-dependent fashion. NSC 95397 also inhibits mitogen-activated protein kinase phosphatase-1 (MKP-1) and suppresses proliferation and induces apoptosis in colon cancer cells through MKP-1 and ERK1/2 pathway.NSC95397 prevents the CtBP1-Protein Partner Interaction and CtBP1-Mediated Transcriptional Repression. NSC-95397 triggers apoptosis of tumor cells and is thus considered for the treatment of malignancy. NSC 95397 is a Cdc25 dual specificity phosphatase inhibitor (Ki=32 nM (Cdc25A), 96 nM (Cdc25B), 40 nM (Cdc25C); IC50=22.3 nM (human Cdc25A), 56.9 nM (human Cdc25C), 125 nM (Cdc25B)).
| Targets |
Mitogen-activated protein kinase phosphatase-1 (MKP-1) (inhibitor, activity-based inhibition, not expression level) [2]
Cdc25 dual-specificity phosphatases (initially identified as selective inhibitor, but in colon cancer cells, the anti-proliferative and pro-apoptotic effects were found to be independent of Cdc25 in this study) [2] |
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| ln Vitro |
In a concentration-dependent way, NSC 95397 (0, 10, and 20 μM; 24 h) decreases the cell survival of clear ducts SW480, SW620, and DLD-1 [2]. Within 24 hours, NSC 95397 (10 μM) folds CDK4 and CDK6, upregulates p21, and affects all bladder waveform line SW480, SW620, and DLD-1 cells [2]. At 10 μM, NSC 95397 decreases the amount of collagenoblast tumor protein on Ser795 and Ser807/811; at 20 μM, NSC 95397 dramatically raises the amounts of internal caspase-9, -3, -7, and PARP [2]. After 6 hours at 10 μM, NSC 95397 improved its downstream.
NSC 95397 reduced cell viability of human colon cancer cell lines SW480, SW620, and DLD-1 in a concentration- and time-dependent manner, as assessed by MTT assay. The IC50 values after 24 h treatment were 9.9 µM for SW480, 14.1 µM for SW620, and 18.6 µM for DLD-1 cells. [2] NSC 95397 suppressed anchorage-independent growth (soft agar colony formation) in SW480, SW620, and DLD-1 cells. Treatment with 10 µM significantly decreased colony formation in SW620 and DLD-1, and nearly abolished it in SW480. At 20 µM, no colonies were formed by any of the three cell lines. [2] NSC 95397 inhibited cell proliferation, as shown by reduced Bromodeoxyuridine (BrdU) incorporation in all three cell lines after 24 h treatment with 10 and 20 µM. [2] NSC 95397 upregulated the expression of the cyclin-dependent kinase inhibitor p21 and downregulated CDK4 and CDK6 protein levels. It also reduced the phosphorylation of retinoblastoma protein (Rb) at Ser795 and Ser807/811 sites. [2] NSC 95397 induced apoptosis in colon cancer cells, as demonstrated by increased Annexin V/7-AAD positive staining (both early and late apoptosis) and elevated levels of cleaved caspase-9, caspase-3, caspase-7, and poly(ADP-ribose) polymerase (PARP). [2] NSC 95397 did not reduce Cdc25A protein level or affect phosphorylation of its downstream target Cdk1 (Tyr15). It inhibited MKP-1 activity (not expression), leading to increased phosphorylation of ERK1/2 (Thr202/Tyr204). The MEK/ERK inhibitor U0126 blocked NSC 95397-induced upregulation of p21 and cleaved caspase-3, confirming the role of the MKP-1/ERK1/2 pathway. [2] |
| Cell Assay |
Cell viability assay [2]
Cell Types: human colon cancer cell line, SW480, SW620 and DLD-1 Tested Concentrations: 0, 10 and 20 μM Incubation times ERK1/ Phosphorylation of 2 at Thr202/Tyr 204[2]. Incubation Duration: 24 hour Experimental Results: IC50 values of NSC 95397 for SW480, SW620 and DLD-1 cell growth were 9.9, 14.1 and 18.6 μM respectively. Western Blot Analysis[2] Cells Lines: SW480, SW620 and DLD-1 Cell Tested Concentrations: 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Upregulation of p21, downregulation of CDK4 and CDK6. diminished phosphorylation of Rb on Ser795 and Ser807/811 Cell Viability Assay (MTT): SW480, SW620, and DLD-1 cells were seeded in 96-well plates. After overnight culture, cells were treated with NSC 95397 at indicated concentrations and times. MTT reagent was added, and optical density was measured after 4 hours to assess cell viability. [2] Anchorage-Independent Growth (Soft Agar Assay): A base layer of 0.6% agar in growth medium was prepared in six-well plates. A top layer of 0.35% agar containing cells was overlaid. Growth medium containing NSC 95397 was added on top and refreshed twice weekly. After four weeks, colonies were stained and counted. [2] Cell Proliferation Assay (BrdU): Cells were seeded in 96-well plates, treated with NSC 95397 for 24 h, and then incubated with BrdU. Incorporation was measured according to the kit protocol. [2] Apoptosis Assay (Flow Cytometry): Cells treated with NSC 95397 for 24 h were harvested, stained with PE Annexin V and 7-AAD, and analyzed by flow cytometry to distinguish live, early apoptotic, and late apoptotic/necrotic cells. [2] Western Blot Analysis: Cells were lysed, proteins were separated by electrophoresis and transferred to membranes. Membranes were blocked and incubated with primary antibodies against p21, CDK4, CDK6, phospho-Rb, caspases, PARP, Cdc25A, phospho-Cdk1, MKP-1, ERK1/2, phospho-ERK1/2, and actin (loading control). After incubation with secondary antibodies, signals were detected. [2] |
| References |
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| Additional Infomation |
2,3-bis(2-hydroxyethylthio)naphthyl-1,4-dione belongs to the class of 1,4-naphthoquinones.
NSC 95397 is a cell-permeable naphthoquinone that was initially identified as a selective inhibitor of the Cdc25 bispecific phosphatase. [2] This study reveals that its antiproliferative and pro-apoptotic effects in colon cancer cells (SW480, SW620, DLD-1) are mediated by the inhibition of MKP-1 activity, leading to ERK1/2 phosphorylation, p21 upregulation, caspase activation, and cell cycle arrest. In this process, the effect is independent of Cdc25. [2] MKP-1 is associated with drug resistance in a variety of cancers, and NSC 95397 has been shown to restore paclitaxel-induced apoptosis in drug-resistant cancer cells by inhibiting MKP-1, suggesting its potential as a treatment to overcome chemotherapy resistance. [2] |
| Molecular Formula |
C14H14O4S2
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|---|---|
| Molecular Weight |
310.38856
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| Exact Mass |
310.033
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| CAS # |
93718-83-3
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| PubChem CID |
262093
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| Appearance |
Light yellow to orange solid powder
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| Density |
1.46g/cm3
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| Boiling Point |
499.2ºC at 760mmHg
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| Flash Point |
255.7ºC
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| LogP |
1.728
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
20
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| Complexity |
381
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
MAASHDQFQDDECQ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H14O4S2/c15-5-7-19-13-11(17)9-3-1-2-4-10(9)12(18)14(13)20-8-6-16/h1-4,15-16H,5-8H2
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| Chemical Name |
2,3-Bis(2-hydroxyethylsulfanyl)naphthalene-1,4-dione
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| Synonyms |
NSC-95397; NSC95397; NSC 95397
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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 |
| 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 : ≥ 100 mg/mL (~322.18 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 10 mg/mL (32.22 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 100.0 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. Solubility in Formulation 2: ≥ 10 mg/mL (32.22 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 100.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2218 mL | 16.1088 mL | 32.2175 mL | |
| 5 mM | 0.6444 mL | 3.2218 mL | 6.4435 mL | |
| 10 mM | 0.3222 mL | 1.6109 mL | 3.2218 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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