| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
Adenosine Monophosphate-Activated Protein Kinase (AMPK) [1]
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|---|---|
| ln Vitro |
KM 91104 (1-10 μM) dose-dependently activated AMPK in human hepatic stellate cells (HSCs), as evidenced by increased phosphorylation of AMPKα (Thr172) and its downstream substrate ACC (Ser79) (western blot): 10 μM increased p-AMPKα by ~2.8-fold and p-ACC by ~3.2-fold compared to control [1]
- It inhibited HSC proliferation: KM 91104 (5 μM, 72 h) reduced HSC viability by ~45% (CCK-8 assay) and blocked cell cycle progression at G0/G1 phase (flow cytometry): G0/G1 phase cells increased from 48% (control) to 72% [1] - Suppression of profibrogenic markers: KM 91104 (10 μM, 24 h) downregulated mRNA expression of α-SMA, collagen I, and fibronectin by ~65%, ~70%, and ~62% respectively (qPCR); corresponding protein levels were reduced by ~58%, ~63%, and ~55% (western blot) [1] - Regulation of vacuolar ATPase (v-ATPase) and intracellular pH: KM 91104 (5 μM) inhibited v-ATPase activity by ~40% (luciferase-based ATPase assay) and normalized intracellular pH (pHi) in activated HSCs (from 6.3 ± 0.1 to 7.1 ± 0.1, pH-sensitive dye detection) [1] - Inhibition of HSC migration: KM 91104 (10 μM) reduced HSC migration by ~52% compared to control (Boyden chamber assay) [1] |
| Enzyme Assay |
AMPK activation assay: Human HSCs were seeded in 6-well plates and cultured to subconfluence. Cells were treated with serial dilutions of KM 91104 (1-10 μM) for 24 h, then lysed in RIPA buffer. Proteins were separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against p-AMPKα (Thr172), total AMPKα, p-ACC (Ser79), and total ACC. Band intensity was quantified by densitometry to assess AMPK activation [1]
- v-ATPase activity assay: Purified v-ATPase from HSCs was resuspended in assay buffer and mixed with KM 91104 (1-10 μM). The reaction mixture was supplemented with ATP and a luciferase-based ATP detection reagent. Luminescence intensity (reflecting remaining ATP) was measured at 0 and 60 minutes, and v-ATPase activity was calculated as the rate of ATP hydrolysis [1] |
| Cell Assay |
HSC proliferation assay: HSCs were seeded in 96-well plates (3×103 cells/well) and cultured overnight. KM 91104 (1-10 μM) was added, and cells were incubated for 72 h. CCK-8 reagent was added, incubated for 2 h, and absorbance was measured at 450 nm to calculate cell viability [1]
- Cell cycle analysis: HSCs were treated with KM 91104 (5 μM) for 72 h, fixed with ethanol, stained with propidium iodide, and analyzed by flow cytometry to determine cell cycle distribution [1] - Profibrogenic marker detection: HSCs were treated with KM 91104 (10 μM) for 24 h. Total RNA was extracted for qPCR analysis of α-SMA, collagen I, and fibronectin mRNA; cell lysates were used for western blot detection of corresponding proteins [1] - Intracellular pH measurement: HSCs were loaded with pH-sensitive fluorescent dye for 30 minutes, then treated with KM 91104 (5 μM) for 1 h. Fluorescence intensity was measured by flow cytometry, and pHi was calculated using a calibration curve [1] - HSC migration assay: HSCs were seeded in the upper chamber of Boyden chambers, and KM 91104 (10 μM) was added to both upper and lower chambers. After 24 h incubation, cells that migrated to the lower membrane were fixed, stained, and counted under a microscope [1] |
| References |
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| Additional Infomation |
KM 91104 is a small molecule AMPK activator that specifically targets the AMPK-vacuole ATPase-pH axis in hepatic stellate cells[1]. Its anti-fibrotic mechanism involves activating AMPK to inhibit v-ATPase activity, normalizing intracellular pH, and thereby inhibiting HSC proliferation, migration, and expression of pro-fibrotic genes/proteins[1]. This compound is considered a potential therapeutic agent for liver fibrosis, and its mechanism of action is through targeting activated hepatic stellate cells, which are a key cell type in the process of liver fibrosis[1].
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| Molecular Formula |
C₁₄H₁₂N₂O₄
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|---|---|
| Molecular Weight |
272.256083488464
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| Exact Mass |
272.08
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| CAS # |
304481-60-5
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| PubChem CID |
135403138
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| Appearance |
White to off-white solid powder
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| LogP |
1.958
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
20
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| Complexity |
361
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC=C(C(=C1)/C=N/NC(=O)C2=CC(=C(C=C2)O)O)O
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| InChi Key |
GWVYHPUGEQGQSF-OVCLIPMQSA-N
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| InChi Code |
InChI=1S/C14H12N2O4/c17-11-4-2-1-3-10(11)8-15-16-14(20)9-5-6-12(18)13(19)7-9/h1-8,17-19H,(H,16,20)/b15-8+
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| Chemical Name |
3,4-Dihydroxybenzoic Acid 2-[(2-Hydroxyphenyl)methylene]hydrazide
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| Synonyms |
KM-91104KM 91104KM91104
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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 (~367.30 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.18 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (9.18 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 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (9.18 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 | 3.6730 mL | 18.3648 mL | 36.7296 mL | |
| 5 mM | 0.7346 mL | 3.6730 mL | 7.3459 mL | |
| 10 mM | 0.3673 mL | 1.8365 mL | 3.6730 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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