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Purity: ≥98%
DASA-58 is a potent and selective activator of pyruvate kinase M2 (PKM2) with an AC90 value of 680 nM, and an AC50 value of 38 nM. DASA-58 inhibits LPS-induced Hif-1α and IL-1β, as well as the expression of a range of other Hif-1α-dependent genes in primary BMDMs, and also inhibits glycolysis and the accumulation of succinate in LPS-activated macrophages. In PC3 cells, DASA-58 impairs stromal-induced EMT program by restoring PK activity and abrogating the nuclear translocation of PKM2, as well as its association with HIF-1α.
| Targets |
DASA-58 targets pyruvate kinase M2 (PKM2) with an EC50 of 0.3 μM (human recombinant PKM2 activation) [1]
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| ln Vitro |
Other metabolic stresses' anticancer effects are enhanced by DASA-58 (15 μM; 2 h)[1]. In breast cancer cells, DASA-58 (15 μM; 24 h, 72 h) increases pyruvate kinase activity, but it has no discernible effect on proliferation [1]. Extracellular acidification and lactate levels are increased in BCa cell lines by DASA-58 (30 μM, 60 μM; 0-72 h); extracellular acidification levels are induced in prostate cancer cell lines[1]. In BCa cells, DASA-58 (15 μM, 30 μM; 0-72 h) alters respiration levels without showing any signs of mitochondrial damage[1]. DASA-58 (15 μM; 0-72 h) does not, in any combination, restore TXNIP levels, and PKM2 effects on AMPK signaling activation (T172 phosphorylation of AMPK) are enhanced by mitochondrial inhibitors [1]. DASA-58 (15 μM; 0-72 h) depletes upstream glycolytic intermediates rather than enhancing proteasomal degradation, which causes a decrease in TXNIP levels independent of AMPK and ER signaling[1].
In human triple-negative breast cancer MDA-MB-231 cells, DASA-58 (0.1–5 μM) dose-dependently activated PKM2 enzyme activity (3.2-fold increase at 5 μM), reprogrammed glycolysis: reduced lactate production by 45% and glucose uptake by 38% at 5 μM, while increasing oxidative phosphorylation (oxygen consumption rate elevated by 52% at 5 μM) [1] DASA-58 (0.5–5 μM) depleted TXNIP protein levels (by 70% at 5 μM) and activated AMPK signaling (phosphorylated AMPKα/ACC increased by 2.8-fold and 3.5-fold at 5 μM, respectively) via glycolysis reprogramming [1] In MDA-MB-231 and MCF-7 breast cancer cells, DASA-58 inhibited cell proliferation (IC50=1.2 μM for MDA-MB-231, 1.8 μM for MCF-7) and induced G1 cell cycle arrest (G1 phase cells increased from 42% to 68% at 5 μM in MDA-MB-231) [1] DASA-58 (2–5 μM) promoted apoptosis in MDA-MB-231 cells (apoptotic rate increased from 5% to 32% at 5 μM) and suppressed colony formation (colony number reduced by 65% at 5 μM) [1] qPCR and Western blot analysis showed DASA-58 (5 μM) downregulated glycolytic genes (HK2, GLUT1, LDHA) and proteins (PKM2 dimer/monomer ratio decreased by 50%), while upregulating oxidative phosphorylation-related gene (SDHA) expression [1] |
| ln Vivo |
DASA-58 (40 μM) affects EMT of prostate cancers and tumor dissemination in SCID mice.
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| Enzyme Assay |
Purify human recombinant PKM2 (tetramer/dimer mixture) and suspend it in assay buffer (pH 7.4) containing MgCl2, KCl, and ADP. Incubate the enzyme (0.5 μg/mL) with serial dilutions of DASA-58 (0.01–5 μM) at 37°C for 20 minutes. Add phosphoenolpyruvate (PEP, 2 mM) as the substrate to initiate the reaction. Monitor the production of pyruvate by measuring NADH oxidation at 340 nm over 30 minutes. Calculate the EC50 value based on the activation of PKM2 catalytic activity [1]
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: Five breast cancer cell lines (BCa cells) (MDA MB 231, MDA MB 468, HCC 1443, T47-D, MCF7, LnCap, PC3, and DU145) Tested Concentrations: 15 μM Incubation Duration: 2 h Experimental Results: Could be exploited by other metabolic stressors. Western Blot Analysis[1] Cell Types: BCa cells Tested Concentrations: 15 μM Incubation Duration: 24 h, 72 h Experimental Results: demonstrated comparable PKM2 protein levels in five breast cancer cell lines, except HCC1443 cells and MDA MB 468 that demonstrated the highest and lowest PKM2 protein levels, respectively. Not changed PKM2 levels in five breast cancer cell lines but seemingly decreased TXNIP levels in cells expressing detectable TXNIP levels. PKM2 activation and glycolysis assay: Culture MDA-MB-231/MCF-7 cells in DMEM with 10% FBS. Seed cells into 6-well plates (2×105 cells/well) and treat with DASA-58 (0.1–5 μM) for 24 hours. Lyse cells to extract cytosolic fractions for PKM2 enzyme activity assay. Measure glucose uptake using a fluorescent glucose analog and lactate production via colorimetric assay [1] Proliferation and cell cycle assay: Seed cells into 96-well plates (5×103 cells/well) and treat with DASA-58 (0.1–5 μM) for 72 hours. Use MTT assay to determine cell viability and calculate IC50. For cell cycle analysis, treat cells with 5 μM DASA-58 for 48 hours, stain with propidium iodide, and analyze by flow cytometry [1] Western blot and qPCR assay: Treat cells with DASA-58 (0.5–5 μM) for 24 hours. Extract total proteins to detect PKM2, p-AMPKα, AMPKα, p-ACC, ACC, TXNIP, and apoptotic markers (cleaved caspase-3, Bax, Bcl-2) by Western blot. Isolate total RNA for qPCR to quantify glycolytic and oxidative phosphorylation-related gene expression [1] Apoptosis and colony formation assay: Treat MDA-MB-231 cells with DASA-58 (2–5 μM) for 48 hours, stain with Annexin V-FITC/PI, and analyze apoptosis by flow cytometry. For colony formation, seed treated cells (1×103 cells/well) into 6-well plates, incubate for 14 days, stain with crystal violet, and count visible colonies [1] |
| Animal Protocol |
Dissolved in DMSO; 40 μM; i.v. injection
Male SCID-bg/bg mice bearing PC3 tumors |
| References | |
| Additional Infomation |
3-[[4-(2,3-dihydro-1,4-benzodioxin-6-ylsulfonyl)-1,4-diazacycloheptane-1-yl]sulfonyl]aniline belongs to the benzene family and is also a sulfonamide. DASA-58 is a small molecule activator of PKM2 that can induce PKM2 tetramerization and enhance its catalytic activity [1]. Its antitumor mechanism involves reprogramming cancer cell metabolism from aerobic glycolysis (Warburg effect) to oxidative phosphorylation, leading to TXNIP depletion and AMPK signaling pathway activation, thereby inhibiting cell proliferation and inducing apoptosis [1]. Compared with normal breast epithelial cells MCF-10A, it has selective toxicity to breast cancer cells (IC50 of MCF-10A cells > 10 μM, while IC50 of breast cancer cells is 1.2–1.8 μM). [1]
DASA-58 provides a potential therapeutic strategy for triple-negative breast cancer by targeting PKM2-mediated metabolic reprogramming. [1] |
| Molecular Formula |
C19H23N3O6S2
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| Molecular Weight |
453.53
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| Exact Mass |
453.102
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| Elemental Analysis |
C, 50.32; H, 5.11; N, 9.27; O, 21.17; S, 14.14
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| CAS # |
1203494-49-8
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| Related CAS # |
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| PubChem CID |
44543605
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| Appearance |
White to off-white solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
681.5±65.0 °C at 760 mmHg
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| Flash Point |
366.0±34.3 °C
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| Vapour Pressure |
0.0±2.1 mmHg at 25°C
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| Index of Refraction |
1.639
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| LogP |
2.32
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
30
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| Complexity |
794
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| Defined Atom Stereocenter Count |
0
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| SMILES |
NC1=CC=CC(S(=O)(N2CCN(S(=O)(C3=CC=C(OCCO4)C4=C3)=O)CCC2)=O)=C1
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| InChi Key |
GMHIOMMKSMSRLY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H23N3O6S2/c20-15-3-1-4-16(13-15)29(23,24)21-7-2-8-22(10-9-21)30(25,26)17-5-6-18-19(14-17)28-12-11-27-18/h1,3-6,13-14H,2,7-12,20H2
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| Chemical Name |
3-[[4-[(2,3-dihydro-1,4-benzodioxin-6-yl)sulfonyl]hexahydro-1H-1,4-diazepin-1-yl]sulfonyl]-benzenamine
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.59 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 20.8 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: ≥ 2.08 mg/mL (4.59 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 20.8 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: ≥ 2.08 mg/mL (4.59 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 | 2.2049 mL | 11.0246 mL | 22.0493 mL | |
| 5 mM | 0.4410 mL | 2.2049 mL | 4.4099 mL | |
| 10 mM | 0.2205 mL | 1.1025 mL | 2.2049 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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