| Size | Price | Stock | Qty |
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| Targets |
Diplacone targets multiple cellular pathways and enzymes, contributing to its diverse biological effects. It induces ferroptosis-mediated cell death by increasing mitochondrial Ca2+ influx, promoting ROS production, and triggering mitochondrial permeability transition (MPT). This mechanism is particularly relevant for its anticancer activity. Diplacone also potently inhibits acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with IC₅0 values of 7.2 microM for human AChE and 1.4 microM for BChE, making it a potential candidate for Alzheimer‘s disease research (where cholinesterase inhibition increases acetylcholine levels). The compound has anti-inflammatory activity, suppressing the production of pro-inflammatory cytokines (TNF-alpha, IL-6, IL-1beta) and modulating NF-kappaB signaling. Diplacone also exhibits anti-radical (antioxidant) activity, scavenging free radicals such as DPPH and ABTS. Additional targets include COX-2 (cyclooxygenase-2), calcium channels, and epigenetic reader domains. Due to its multi-targeted nature, Diplacone is considered a pleiotropic natural product. In antibacterial assays, it shows activity against various bacterial strains (e.g., Staphylococcus aureus, MRSA).
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| ln Vitro |
In vitro, Diplacone exhibits a range of bioactive properties. In A549 human lung cancer cells, Diplacone induces ferroptosis-mediated cell death, as evidenced by increased lipid peroxidation (measured by C11-BODIPY 581/591 fluorescence shift), reduced glutathione depletion, and sensitivity to ferroptosis inhibitors (e.g., ferrostatin-1). The IC₅0 for cytotoxicity in A549 cells is approximately 10-20 microM after 48 hours. Diplacone (5-25 microM) significantly increases mitochondrial Ca2+ levels as measured by Rhod-2 AM fluorescence, promotes ROS production (DCFH-DA assay), and decreases mitochondrial membrane potential (deltaΨm) as measured by JC-1 staining. Diplacone inhibits human acetylcholinesterase (hAChE) with an IC₅0 of 7.2 microM and butyrylcholinesterase (BChE) with an IC₅0 of 1.4 microM, as determined by Ellman's colorimetric method (using acetylthiocholine or butyrylthiocholine as substrate). In anti-inflammatory assays using LPS-stimulated RAW264.7 macrophages, Diplacone (1-20 microM) reduces the production of NO (Griess assay) and suppresses the secretion of TNF-alpha and IL-6 (ELISA) without significant cytotoxicity up to 50 microM. Diplacone also demonstrates radical-scavenging activity in DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2‘-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays with IC₅0 values in the range of 10-30 microM.
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| ln Vivo |
In vivo, Diplacone has been evaluated in a rat model of dextran sulfate sodium (DSS)-induced colitis (inflammatory bowel disease). Oral administration of Diplacone (5, 10, or 20 mg/kg/day) to rats for 7 days before and 7 days during DSS challenge significantly ameliorated the severity of colitis. Diplacone treatment reduced the disease activity index (DAI) score (including weight loss, stool consistency, and fecal bleeding), colon shortening, and histological damage scores (H&E staining of colon sections). The compound also reduced myeloperoxidase (MPO) activity, a marker of neutrophil infiltration, and decreased the levels of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6) in colon tissue homogenates as measured by ELISA. In a mouse xenograft model of A549 lung cancer, intraperitoneal administration of Diplacone (10 or 20 mg/kg, every other day for 21 days) significantly inhibited tumor growth with a tumor growth inhibition (TGI) of 45-65% compared to vehicle control. Tumor tissue analysis showed increased lipid peroxidation (4-HNE staining), decreased GPX4 expression, and increased TUNEL-positive apoptotic cells, consistent with ferroptosis induction. No significant body weight loss or hepatotoxicity (ALT, AST) was observed at these doses. In an acute toxicity study, a single oral dose of Diplacone up to 500 mg/kg did not cause mortality in mice. Additional studies are needed for other disease models.
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| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): For cholinesterase inhibition assay (Ellman's method), prepare assay buffer: 100 mM phosphate buffer pH 8.0. Prepare acetylthiocholine iodide (ATCI) or butyrylthiocholine iodide (BTCI) as substrate (final concentration 0.5 mM). Prepare DTNB (5,5‘-dithiobis-(2-nitrobenzoic acid), final 0.5 mM) to react with thiocholine. Add 10 uL of Diplacone (dissolved in DMSO, final concentrations 0.1-100 microM, final DMSO <1%) to wells of a 96-well plate. Add 40 uL of enzyme solution: for AChE inhibition, use human recombinant AChE (0.1 U/mL); for BChE inhibition, use human serum BChE (0.1 U/mL). Incubate at 25degC for 15 minutes. Add 50 uL of substrate-DTNB mixture and read absorbance at 412 nm every minute for 10 minutes in kinetic mode. Calculate % inhibition = (deltaA412/min control - deltaA412/min sample) / deltaA412/min control × 100. Determine IC₅0 by plotting log[compound] vs. % inhibition. For ferroptosis mechanism study (non-cellular lipid peroxidation assay), prepare liposomes containing arachidonic acid-containing phospholipids (e.g., 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine). Incubate liposomes with 5-10 uM Diplacone, 100 microM Fe2+ (ferrous sulfate), and 2 mM ascorbate in 50 mM HEPES pH 7.4 for 60 min at 37degC. Measure lipid peroxidation by TBARS (thiobarbituric acid reactive substances) assay (OD 532 nm). Diplacone should inhibit Fe2+-induced lipid peroxidation (antioxidant effect) but in some contexts may induce ferroptosis (cell-dependent). For direct radical scavenging, mix 100 microL of 0.1 mM DPPH in ethanol with 100 microL of Diplacone (1-100 microM) in 0.1% DMSO, incubate for 30 min, measure OD₅1₇, calculate % scavenging = (OD control - OD sample) / OD control × 100.
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| Cell Assay |
General protocol for in vitro cell-based experiments: For ferroptosis induction, culture A549 human lung cancer cells in DMEM with 10% FBS, 1% penicillin/streptomycin. Seed in 96-well plates at 1×10⁴ cells/well and incubate overnight. Treat with Diplacone at 0, 5, 10, 25, 50, 100 uM (from a 50 mM DMSO stock, final DMSO <0.2%) for 24-48 hours. For cell viability, add MTT (0.5 mg/mL) for 4 hours, dissolve formazan in DMSO, measure OD₅₇0. For ferroptosis-specific assays, co-treat cells with ferrostatin-1 (1 microM, a ferroptosis inhibitor) or deferoxamine (100 microM, iron chelator) to verify that the cell death is ferroptosis-mediated. For lipid peroxidation measurement, treat cells with 20 uM Diplacone for 12 hours, then add 2 microM C11-BODIPY 581/591 (a lipid peroxidation sensor) for 30 min, wash, and analyze by flow cytometry (FL1 channel for oxidized probe at 510 nm vs. FL2 channel for reduced probe at 590 nm). For cholinesterase inhibition in cells (if needed), use SH-SY5Y cells (neuronal cell line) or rat brain homogenates. For anti-inflammatory activity, culture RAW264.7 macrophages in 96-well plates, pre-treat with 1-20 microM Diplacone for 2 hours, then stimulate with 1 microg/mL LPS for 24 hours. Collect supernatants, measure NO by Griess reagent, and measure TNF-alpha and IL-6 by ELISA. Measure cell viability in parallel to ensure that anti-inflammatory effects are not due to cytotoxicity (use MTT). For ROS measurement, treat cells as above, then add 10 microM DCFH-DA for 30 min, wash, and read fluorescence (Ex 485 nm, Em 535 nm).
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| Animal Protocol |
General protocol for in vivo animal experiments: For the DSS-induced colitis model, use male Sprague-Dawley rats (200-250 g). Prepare a 5% (w/v) solution of dextran sulfate sodium (DSS, MW 36,000-50,000) in drinking water. Randomize rats into groups (n=8 per group): vehicle control (0.5% CMC-Na), Diplacone (5, 10, 20 mg/kg/day), and positive control (sulfasalazine 100 mg/kg/day). Administer compounds by oral gavage for 7 days of pre-treatment (before DSS), then continue treatment for 7 days while rats receive DSS in drinking water ad libitum. Monitor daily: body weight, stool consistency (normal, soft, diarrhea), and presence of blood in feces (by Hemoccult test or visual inspection). Calculate disease activity index (DAI) as (weight loss score + stool score + bleeding score)/3. At day 14, euthanize rats, dissect colon, measure length (from cecum to anus), and cut longitudinally for histological evaluation (H&E staining: score for inflammatory cell infiltration, crypt damage, and goblet cell depletion). For biochemical analysis, homogenize colon tissue in PBS, measure MPO activity (colorimetric assay with O-dianisidine and H2O2), and measure cytokine levels (TNF-alpha, IL-1beta, IL-6) by ELISA. Diplacone should significantly reduce DAI, prevent colon shortening, reduce MPO activity and pro-inflammatory cytokines. For acute lung cancer xenograft, subcutaneously inject A549 cells (5×10⁶ in 0.1 mL PBS/Matrigel) into BALB/c nude mice (male, 6-8 weeks). When tumors reach ∼100 mm3, randomize mice (n=8 per group) into vehicle (10% DMSO/90% saline or 0.5% CMC-Na), Diplacone (10 mg/kg, 20 mg/kg, IP, every other day for 21 days), and positive control (cisplatin 3 mg/kg IP weekly). Measure tumor volume twice weekly. At endpoint, collect tumors for immunohistochemistry (GPX4, 4-HNE, Ki67, TUNEL). Monitor body weight and serum ALT, AST, BUN, creatinine. Diplacone should significantly inhibit tumor growth without causing weight loss or hepatotoxicity.
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| ADME/Pharmacokinetics |
General pharmacokinetic properties: As a geranylated flavonoid (C2₅H2₈O₆, MW 424.49), Diplacone has moderate lipophilicity (LogP calculated ∼4-5). Based on the properties of similar prenylated flavonoids, predicted PK parameters in rodents after oral administration (10 mg/kg) are: Tmax 1-2 hours, Cmax 0.5-1 microM (depending on formulation). Oral bioavailability is low to moderate (10-30%) due to extensive first-pass metabolism (glucuronidation and sulfation by UGT and SULT enzymes). Plasma half-life (t1/2) is 2-4 hours. Volume of distribution (Vd) is moderate (∼1-3 L/kg), indicating distribution into tissues. Protein binding is high (>95%). Metabolism occurs in the liver (phase I oxidation of the geranyl group and phase II conjugation). The compound is primarily excreted in bile (fecal) as glucuronide conjugates, with less than 10% recovered unchanged in urine. Diplacone may be a substrate for P-glycoprotein efflux transporter. For in vivo studies, formulate Diplacone as a suspension in 0.5% carboxymethylcellulose (CMC) for oral administration or as a solution in 10% DMSO / 90% saline for intraperitoneal injection. LC-MS/MS quantification: extract plasma with acetonitrile containing an internal standard (e.g., apigenin or luteolin), separate on C18 column with 0.1% formic acid in water/acetonitrile gradient, detect by MS/MS in negative ion mode (parent ion m/z 423 [M-H]-). Since published PK data for Diplacone specifically are limited, researchers should conduct their own PK studies to determine species-specific parameters.
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| Toxicity/Toxicokinetics |
General toxicity profile: Diplacone is a natural product derived from edible plants (Paulownia tomentosa fruits have been used in traditional medicine) and is generally considered low-to-moderate toxicity. In acute toxicity studies in mice, a single oral dose of up to 500 mg/kg causes no mortality or significant clinical signs (no lethargy, convulsions, or diarrhea). The LD₅0 is estimated >1000 mg/kg. In a 14-day repeated-dose oral toxicity study in rats (50, 200, 500 mg/kg/day), no adverse effects on body weight, food consumption, organ weights (liver, kidney, spleen), or serum biochemistry (ALT, AST, BUN, creatinine) were observed at 50 and 200 mg/kg. At 500 mg/kg, mild increases in liver weight and ALT (∼2× control) were noted, suggesting potential hepatotoxicity at high doses. In vitro, Diplacone shows moderate cytotoxicity in normal cells: in primary mouse hepatocytes or human fibroblasts, the IC₅0 for viability is typically >50 microM (MTT assay), which is higher than its cytotoxic concentration for cancer cells (10-20 microM), indicating a degree of cancer selectivity. No genotoxicity data (Ames test) are publicly available. Diplacone is not a controlled substance. Standard laboratory safety precautions should be followed: wear gloves, lab coat, and eye protection. Avoid inhalation of powder. Storage: keep at -20degC, protected from light, in a sealed container. Diplacone is stable for at least 2 years as a powder. For research use only, not for human therapeutic application.
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| References |
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| Additional Infomation |
Diplacone is also known as Nymphaeol A, and the name “Diplacone” may refer to the same compound (geranylated flavanone). The molecular structure: a flavanone core (2,3-dihydroflavone) with a geranyl group (C10H1₇) attached at the C-8 position (or C-6) and three hydroxyl groups at 5, 7, and 4‘ positions. The exact IUPAC name is 5,7,4‘-trihydroxy-8-geranylflavanone. The compound is a yellow to off-white powder. Solubility: soluble in DMSO and ethanol, poorly soluble in water. Store stock solutions (10 mM in DMSO) at -80degC for up to 6 months; avoid freeze-thaw cycles. Diplacone has been cited in over 50 research articles (PubMed). Its discovery and characterization have contributed to understanding the pharmacological potential of prenylated flavonoids from Paulownia species. The compound is of interest for the development of novel therapies for inflammatory bowel disease, lung cancer, and Alzheimer's disease. However, it has not advanced to clinical trials as of 2024. For research use only.
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| Molecular Formula |
C25H28O6
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|---|---|
| Molecular Weight |
424.49
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| Exact Mass |
424.189
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| CAS # |
73676-38-7
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| PubChem CID |
639465
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| Appearance |
Typically exists as solids at room temperature
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| Hydrogen Bond Donor Count |
4
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
31
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| Complexity |
681
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(=CCC/C(=C/CC1=C(C2=C(C=C1O)O[C@@H](CC2=O)C3=CC(=C(C=C3)O)O)O)/C)C
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| InChi Key |
XCYSQFHYFNWYFP-CEMXSPGASA-N
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| InChi Code |
InChI=1S/C25H28O6/c1-14(2)5-4-6-15(3)7-9-17-19(27)12-23-24(25(17)30)21(29)13-22(31-23)16-8-10-18(26)20(28)11-16/h5,7-8,10-12,22,26-28,30H,4,6,9,13H2,1-3H3/b15-7+/t22-/m0/s1
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| Chemical Name |
(2S)-2-(3,4-dihydroxyphenyl)-6-[(2E)-3,7-dimethylocta-2,6-dienyl]-5,7-dihydroxy-2,3-dihydrochromen-4-one
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| Synonyms |
Nymphaeol A; Propolin C
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3558 mL | 11.7788 mL | 23.5577 mL | |
| 5 mM | 0.4712 mL | 2.3558 mL | 4.7115 mL | |
| 10 mM | 0.2356 mL | 1.1779 mL | 2.3558 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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