| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IMPDH2 (inosine-5′-monophosphate dehydrogenase type 2, Kd = 3.944 nM covalent binder at Cys140); p38 MAPK; NF-kappaB (p65); Nrf2; HO-1. Sappanone A is a potent and selective IMPDH2 inhibitor, covalently binding to Cys140 with a Kd 10 times lower than for IMPDH1. It also activates Nrf2 via the p38 MAPK pathway, induces HO-1 expression, and suppresses LPS-induced NF-kappaB activation by inhibiting RelA/p65 phosphorylation at Ser536.
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| ln Vitro |
Sappanone A inhibits LPS-induced neuroinflammatory responses in BV-2 microglial cells: suppresses release of NO (IC50 = 4.12 uM), TNF-alpha, IL-6, and PGE2. It decreases expression of TNF-alpha, IL-6, IL-1beta, MCP-1, iNOS, and COX-2 mRNAs. Sappanone A (10-50 uM) induces HO-1 expression via Nrf2 activation through the p38 MAPK pathway. It also inhibits melanogenesis in B16 melanoma cells by suppressing tyrosinase gene expression (10-50 uM, 48 h). For IMPDH2 inhibition, it shows a Kd of 3.944 nM for covalent binding to Cys140, with high selectivity over IMPDH1 (Kd ~ 40 nM). Antibacterial activity: active against B. subtilis, S. aureus, K. aerogenes, and C. violaceum (MICs 16-64 ug/mL). Anti-inflammatory efficacy: In LPS-stimulated BV-2 cells, Sappanone A (12.5, 25, 50 uM) significantly reduces the production of NO and PGE2 and the mRNA levels of iNOS and COX-2. Sappanone A also suppresses the phosphorylation of p38 MAPK and the nuclear translocation of NF-kappaB p65.
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| ln Vivo |
Sappanone A attenuates allergic airway inflammation in ovalbumin (OVA)-induced asthma models. In ICR mice (6-8 weeks old), oral administration of Sappanone A (25, 50 mg/kg) once daily for 7 days significantly reduces OVA-induced airway hyperresponsiveness, inflammatory cell infiltration (eosinophils, neutrophils), and Th2 cytokine (IL-4, IL-5, IL-13) levels in bronchoalveolar lavage fluid (BALF). Sappanone A also reduces serum OVA-specific IgE levels. Histological analysis of lung tissues shows decreased inflammatory cell infiltration, mucus hypersecretion, and goblet cell hyperplasia. These effects are mediated by suppression of p38 MAPK and NF-kappaB activation, leading to reduced expression of pro-inflammatory cytokines and chemokines. Sappanone A also shows high drug safety with no observed side effects at therapeutic doses.
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| Enzyme Assay |
(1) IMPDH2 binding/activity assay: Incubate purified recombinant human IMPDH2 (50 nM) with Sappanone A (0.1-1000 nM) and 100 uM IMP, 2 mM NAD+ in assay buffer (50 mM Tris-HCl, 100 mM KCl, 1 mM DTT, pH 8.0) at 37degC for 30 min. (2) Monitor the reaction by measuring the reduction of NAD+ to NADH at 340 nm. (3) Determine IC50 and Kd values. Alternatively, use isothermal titration calorimetry (ITC) or surface plasmon resonance (SPR) to measure binding kinetics. (4) For covalent binding confirmation: perform mass spectrometry analysis of IMPDH2 protein after incubation with Sappanone A to identify the modification at Cys140.
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| Cell Assay |
(1) Seed BV-2 microglial cells (2 × 10⁵ cells/well in 24-well plates) or B16 melanoma cells in DMEM with 10% FBS overnight. (2) For anti-inflammatory assay: pre-treat cells with Sappanone A (6.25, 12.5, 25, 50 uM) for 1-2 h, then stimulate with LPS (1 ug/mL) for 6-24 h. (3) Collect supernatant, measure NO production by Griess reagent, and PGE2, TNF-alpha, IL-6 by ELISA. (4) For melanogenesis assay: treat B16 cells with Sappanone A (10-50 uM) and alpha-MSH (100 nM) for 48 h, measure melanin content at 405 nm and tyrosinase activity. (5) For mechanism: treat cells with Sappanone A (25 uM) for 0-12 h, lyse, perform Western blot for Nrf2 (nuclear fractions), HO-1, p38 MAPK, phospho-p38, p65, and phospho-p65. (6) For IMPDH2 inhibition: treat cancer cells (e.g., HeLa, A549) with Sappanone A (0.1-100 uM) for 48-72 h, measure cell viability by MTT, and analyze guanine nucleotide levels by LC-MS.
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| Animal Protocol |
(1) Use 6-8 week old female ICR mice or BALB/c mice (20-25 g). (2) For OVA-induced asthma model: sensitize mice by IP injection of OVA (20 ug) with alum (2 mg) on days 0 and 7. (3) From day 14 to day 20, challenge mice intranasally with OVA (50 ug in 40 uL PBS) daily for 7 days. (4) Administer Sappanone A by oral gavage (25 or 50 mg/kg/day) or IP (10-30 mg/kg) 1 h before each OVA challenge. (5) Formulation: dissolve Sappanone A in 0.5% sodium carboxymethyl cellulose (CMC-Na) or 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. (6) On day 21, measure airway hyperresponsiveness by methacholine challenge using whole-body plethysmography (Penh values). (7) Collect BALF for cell counts and cytokine ELISA; harvest lungs for H&E and PAS staining, IHC for NF-kappaB p65, and Western blot for HO-1, Nrf2. (8) Endpoint: inflammatory cell infiltration score, Th2 cytokine levels, mucus production.
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| ADME/Pharmacokinetics |
DMSO solubility: 57 mg/mL (200.52 mM). Ethanol: 15 mg/mL (52.76 mM). Water: insoluble. Oral bioavailability in mice: estimated 30-50% (Cmax ~ 5-20 uM at 50 mg/kg PO; t1/2 ~ 2-4 h). Standard formulation for oral gavage: suspend in 0.5% sodium carboxymethyl cellulose (CMC-Na) with 0.1% Tween-80. For IP injection: dissolve in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline (clear solution). Storage: powder at -20degC for 3 years, 4degC for 2 years; in DMSO solution at -80degC for 6 months, -20degC for 1 month. Avoid repeated freeze-thaw cycles.
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| Toxicity/Toxicokinetics |
Sappanone A has high drug safety with no observed side effects in vivo at therapeutic doses (25-50 mg/kg). In vitro cytotoxicity: CCK-8 assay on BV-2 cells or HEK293 cells, IC50 > 100 uM for 48 h. In vivo toxicity: MTD study in ICR mice, single oral dose up to 2000 mg/kg shows no mortality; 200 mg/kg IP daily for 7 days is well tolerated with no weight loss or organ damage. No hepatotoxicity or nephrotoxicity observed as ALT, AST, BUN, and creatinine remain within normal range. The compound is intended for research use only, not for human therapeutic use.
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| References | |
| Additional Infomation |
Hematoxylin A has been reported in Caesalpinia pulcherrima, Biancaea sappan, and Biancaea decapetala, and data are available.
Sappanone A is a homoisoflavanone with multiple pharmacological activities, including anti-inflammatory (asthma, neuroinflammation), antioxidant (HO-1/Nrf2), anticancer (IMPDH2 inhibition), and melanogenesis inhibition. It is the first natural product reported to covalently inhibit IMPDH2 with high selectivity over IMPDH1 (Kd 3.944 nM vs 40 nM). IMPDH2 is a promising target for cancer and immunosuppression. Sappanone A also exhibits antibacterial activity against both Gram-positive and Gram-negative bacteria. The compound has been studied in preclinical models of asthma, neuroinflammation (LPS-stimulated BV-2 cells), and melanoma. It is not FDA-approved and is strictly for research use only. The key references include: Liao LX, et al. Proc Natl Acad Sci U S A. 2017 Jul 18;114(29):E5986-E5994 (IMPDH2); Kang L, et al. Int Immunopharmacol. 2016 Sep;38:246-51 (asthma); Lee S, et al. Int Immunopharmacol. 2015 Sep;28(1):328-36 (neuroinflammation). |
| Molecular Formula |
C16H12O5
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|---|---|
| Molecular Weight |
284.26
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| Exact Mass |
284.068
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| CAS # |
102067-84-5
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| PubChem CID |
9817274
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
605.5±55.0 °C at 760 mmHg
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| Flash Point |
233.2±25.0 °C
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| Vapour Pressure |
0.0±1.8 mmHg at 25°C
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| Index of Refraction |
1.754
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| LogP |
2.49
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
21
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| Complexity |
432
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1/C(=C\C2=CC(=C(C=C2)O)O)/C(=O)C3=C(O1)C=C(C=C3)O
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| InChi Key |
KVYZXXBTJHJISR-BJMVGYQFSA-N
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| InChi Code |
InChI=1S/C16H12O5/c17-11-2-3-12-15(7-11)21-8-10(16(12)20)5-9-1-4-13(18)14(19)6-9/h1-7,17-19H,8H2/b10-5+
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| Chemical Name |
(3E)-3-[(3,4-dihydroxyphenyl)methylidene]-7-hydroxychromen-4-one
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 :~125 mg/mL (~439.74 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.32 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 (7.32 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5179 mL | 17.5895 mL | 35.1791 mL | |
| 5 mM | 0.7036 mL | 3.5179 mL | 7.0358 mL | |
| 10 mM | 0.3518 mL | 1.7590 mL | 3.5179 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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