| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| Targets |
Liver kinase B1 (LKB1) - Phillyrin induces phosphorylation of LKB1 at Ser428, leading to AMPK activation. [3]
AMP-activated protein kinase (AMPK) - Phillyrin promotes AMPKα phosphorylation at Thr172. [3] Cytochrome P450 isoforms: CYP1A2 (induction), CYP2D1 (induction) - no IC50 values reported. [1] |
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| ln Vitro |
Phillyrin (1–5 μM; pretreatment 1 hour; 24 hours) can considerably reduce the amount of FAS protein that is caused by high glucose in HepG2 cells [1]. Human HepG2 hepatocytes activated SREBP-1c is inhibited by phillyrin (1–5 μM; 1 hour pretreatment; 24 hours) to prevent high glucose-induced FAS mRNA expression [1].
In human HepG2 hepatocytes, Phillyrin (1-5 µM) significantly suppressed high glucose (30 mM)-induced lipid accumulation. Nile red staining showed that 1, 2.5, and 5 µM Phillyrin reduced lipid accumulation by 15.5%, 26.3%, and 35.7% respectively. Triglyceride levels were reduced by 22.8%, 45.1%, and 60.3%; total cholesterol content was decreased by 20.7%, 31.6%, and 52.5% respectively. [3] Phillyrin (1-5 µM) inhibited high glucose-induced fatty acid synthase (FAS) protein and mRNA expression in HepG2 cells as measured by Western blotting and qRT-PCR. [3] Phillyrin reduced high glucose-induced accumulation of mature SREBP-1 protein in the nucleus (Western blotting) and decreased SREBP-1c mRNA expression (qRT-PCR). It also inhibited SREBP-1c promoter activity in a luciferase reporter assay. [3] Phillyrin (1-5 µM) stimulated phosphorylation of AMPKα at Thr172 and ACC at Ser79 under normal glucose (5.5 mM) and restored their phosphorylation under high glucose conditions (30 mM) in HepG2 cells, as determined by Western blotting. The AMPK inhibitor compound C (10 µM) reversed the inhibitory effects of Phillyrin on SREBP-1 and FAS expression. [3] Phillyrin (5 µM) induced time-dependent phosphorylation of LKB1 at Ser428 (maximal at 0.5 h) and AMPKα at Thr172 (maximal at 3 h) in HepG2 cells. In LKB1-deficient HeLa cells, Phillyrin failed to increase AMPKα Thr172 phosphorylation or reduce FAS and mature nuclear SREBP1. STO-609 (CaMKβ inhibitor) did not abolish Phillyrin-induced AMPKα phosphorylation. [3] In rat liver microsomes from Phillyrin-pretreated rats (10 mg/kg/day for 7 days), the concentration of acetaminophen (CYP1A2 metabolite) was increased by 207.69% (p<0.01) and dextrophan (CYP2D1 metabolite) by 125.00% (p<0.05) compared to control, indicating induction of CYP1A2 and CYP2D1. No significant changes were observed for 4-hydroxytolbutamide (CYP2C11) or 6β-hydroxytestosterone (CYP3A1/2). [1] |
| ln Vivo |
In male BALB/c mice infected with influenza A virus (A/FM/1/47 strain, 4 LD50 intranasally), Phillyrin administered intraperitoneally at 20 mg/kg/day for 3 days significantly prolonged mean survival time (8.7 days vs. 6.5 days in virus control group) as analyzed by Kaplan-Meier method (p<0.05). Survival rates were measured daily for 14 days. Body weights did not differ significantly among groups. [2]
Phillyrin (20 mg/kg/day, i.p., 3 days) significantly reduced virus titers in lung homogenates on day 5 post-infection (plaque reduction assay) and decreased lung index (lung weight/body weight) compared to virus-infected group. Western blotting showed reduced influenza HA protein expression, and qRT-PCR showed decreased M gene mRNA expression (not statistically significant). [2] Phillyrin (20 mg/kg/day) significantly decreased serum levels of IL-6 and GM-CSF on day 5 post-infection (Bio-Plex assay) compared to virus-infected group. IL-12 and IFN-γ levels were also decreased but not significantly. Histopathological examination (H&E staining) of lung tissue on day 5 showed reduced inflammation and consolidation in Phillyrin-treated mice compared to virus controls. [2] In male Wistar rats, Phillyrin pretreatment (10 mg/kg/day intraperitoneally for 7 days) accelerated the metabolism of caffeine (CYP1A2 substrate) and metoprolol (CYP2D1 substrate) in vivo. Cmax and AUC0-∞ of caffeine decreased by 22.15% (p<0.01) and 118.19% (p<0.05) respectively; Cmax and AUC0-∞ of metoprolol decreased by 22.28% (p<0.05) and 22.91% (p<0.01) respectively. No significant changes were observed for tolbutamide (CYP2C11) or dapsone (CYP3A1/2). [1] |
| Enzyme Assay |
Rat liver microsomes (RLM) were prepared by differential centrifugation from rats pretreated with Phillyrin (10 mg/kg/day i.p. for 7 days). Incubation mixtures contained microsomal protein (1 mg/mL for phenacetin O-deethylation, tolbutamide 4-hydroxylation, dextromethorphan O-demethylation; 0.65 mg/mL for testosterone 6β-hydroxylation), NADPH regenerating system (NADPH 1 mM, MgCl2 1 mM), and probe substrates (phenacetin 25 µM for CYP1A2, tolbutamide 200 µM for CYP2C11, dextromethorphan 5 µM for CYP2D1, testosterone 50 µM for CYP3A1/2) in 0.1 M potassium phosphate buffer pH 7.4. After preincubation at 37°C for 5 min, reaction started by adding NADPH. Incubation times: 60 min for phenacetin, tolbutamide, dextromethorphan; 45 min for testosterone. Reactions terminated with ice-cold acetonitrile, then internal standards added, centrifuged (9000g, 15 min), and supernatant analyzed by HPLC. Metabolites measured: acetaminophen (CYP1A2), 4-hydroxytolbutamide (CYP2C11), dextrophan (CYP2D1), 6β-hydroxytestosterone (CYP3A1/2). Phillyrin pretreatment significantly increased acetaminophen and dextrophan concentrations (207.69% and 125.00% increase respectively) compared to control. [1]
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| Cell Assay |
RT-PCR[1]
Cell Types: Human HepG2 hepatocyte Tested Concentrations: 1 μM; 2.5 μM; 5 μM Incubation Duration: 24 hrs (hours) Experimental Results: FAS and SREBP-1c mRNA expression diminished. HepG2 cells were cultured in DMEM with normal glucose (5.5 mM) and 10% FBS. For high glucose-induced lipid accumulation, cells were serum-starved overnight then incubated with 30 mM D-glucose for 24 h. Phillyrin (1-5 µM) was added 1 h before high glucose exposure. Nile red staining: cells fixed with 4% paraformaldehyde for 30 min, stained with 1 µg/mL Nile red for 10 min in dark, fluorescence detected at ex 488 nm/em 550 nm. Triglyceride and total cholesterol: cell homogenates extracted with chloroform:methanol (2:1), organic phase evaporated, pellet dissolved in PBS with 1% Triton X-100, measured using enzymatic kits at 550 nm and 500 nm, normalized to protein. [3] Western blotting: cells lysed in RIPA buffer, proteins separated by SDS-PAGE (8-10%), transferred to PVDF membrane, blocked with 5% skim milk, incubated with primary antibodies against FAS, SREBP-1, p-AMPKα (Thr172), AMPKα, p-ACC (Ser79), p-LKB1 (Ser428), LKB1, Lamin B1, β-actin, then HRP-conjugated secondary antibodies, visualized by ECL. Nuclear and cytoplasmic extracts prepared using hypotonic buffer and Buffer C. [3] qRT-PCR: total RNA isolated using RNAiso plus, cDNA synthesized, PCR performed with SYBR Green, primers for SREBP-1c, FAS, β-actin. mRNA expression normalized to β-actin using comparative Ct method. [3] Luciferase assay: cells transfected with SREBP-1c promoter-luciferase plasmid and pCMV-β-gal using Lipofectamine 2000. After 5 h, cells treated with Phillyrin (1-5 µM) for 24 h, lysed, luciferase activity measured by luminometer, normalized to β-galactosidase activity. [3] RNA interference: LKB1 siRNA or scrambled siRNA transfected using oligofectamine. [3] Cell viability and cytotoxicity: MTT assay (0.5 mg/mL, 1 h, absorbance 570 nm) and LDH release assay (absorbance 490 nm) in HepG2 cells treated with Phillyrin (1-100 µM) for 24 h. Phillyrin showed no cytotoxicity or effect on cell viability. [3] |
| Animal Protocol |
For influenza A virus infection study: Male BALB/c mice (17-19 g) were anesthetized with 1% pentobarbital (50 mg/kg i.p.) and infected intranasally with 20 µL of influenza A virus A/FM/1/47 strain at 4 LD50 diluted in PBS. 24 h post-infection, mice were treated with Phillyrin (10 or 20 mg/kg/day) intraperitoneally for 3 consecutive days. Control groups: normal control (saline), virus control (saline), oseltamivir (10 mg/kg/day oral gavage). Survival rates and body weights were observed for 14 days. On day 5 post-infection, lungs were removed for lung index calculation (lung weight/body weight), virus titer by plaque assay, HA protein by Western blot, M gene expression by qRT-PCR, and histopathology (H&E staining). Serum cytokines (IL-6, GM-CSF, IL-12, IFN-γ) measured on days 3 and 5 using Bio-Plex assay. [2]
For CYP450 interaction study: Male Wistar rats (200±20 g) were randomly divided into blank control (0.9% NaCl i.p. for 7 days), Phillyrin pretreatment (10 mg/kg/day i.p. for 7 days), and forsythoside A pretreatment groups. On day 8, all rats received a cocktail solution (5 mL/kg) containing caffeine (10 mg/kg for CYP1A2), tolbutamide (10 mg/kg for CYP2C11), metoprolol (20 mg/kg for CYP2D1), and dapsone (10 mg/kg for CYP3A1/2) intraperitoneally. Blood samples (300 µL) collected from tail vein at 0, 0.17, 0.5, 0.83, 1.17, 1.5, 2, 3, 5, 8, 12, 24 h. Plasma separated by centrifugation (4000g, 10 min), stored at -20°C. Probe drugs analyzed by UHPLC-MS-MS. Pharmacokinetic parameters calculated using DAS 2.0 software. [1] For microsome preparation: After 7-day pretreatment, rats were anesthetized with 4% halothane, livers removed (fasted 24 h), washed with cold 0.9% NaCl, and liver microsomes prepared by differential centrifugation. Protein concentration determined by BCA assay. Microsomes stored at -80°C. [1] |
| ADME/Pharmacokinetics |
In rats, Phillyrin pretreatment (10 mg/kg/day i.p. for 7 days) significantly altered pharmacokinetics of CYP1A2 and CYP2D1 probe substrates. For caffeine (CYP1A2 substrate): Cmax decreased by 22.15% (p<0.01), AUC0-∞ decreased by 118.19% (p<0.05). For metoprolol (CYP2D1 substrate): Cmax decreased by 22.28% (p<0.05), AUC0-∞ decreased by 22.91% (p<0.01). No significant changes in tolbutamide or dapsone pharmacokinetics. [1]
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| Toxicity/Toxicokinetics |
In HepG2 cells, Phillyrin (1-100 µM for 24 h) showed no cytotoxicity as measured by LDH release assay and did not affect cell viability as measured by MTT assay under both normal glucose (5.5 mM) and high glucose (30 mM) conditions. [3]
In male Wistar rats, intraperitoneal administration of Phillyrin at 10 mg/kg/day for 7 days was well-tolerated with no reported adverse effects. [1] In male BALB/c mice, Phillyrin at 20 mg/kg/day i.p. for 3 days did not cause significant body weight loss compared to virus-infected controls. [2] |
| References |
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| Additional Infomation |
Phillyrin is a natural lignan found in Forsythia suspensa (Oleaceae). It has anti-obesity activity in nutritive obesity mice by lowering fat weight, fat index, fat cell diameter, Lee's index, jejunum microvillus area, and serum triglycerides and cholesterol. [3]
Phillyrin shows anti-inflammatory activity by attenuating lipopolysaccharide-induced inflammation. It has antibacterial and antioxidant activities. [2] Phillyrin is a major component of traditional Chinese medicine preparations such as Shuanghuanglian injection and Tanreqing injection, often used in combination with other drugs (e.g., azithromycin for mycoplasmal pneumonia). [1] Phillyrin induces CYP1A2 and CYP2D1 in rats, which may accelerate the metabolism of drugs that are substrates of these enzymes (e.g., theophylline, metoprolol, tricyclic antidepressants, β-blockers). This could lead to reduced plasma concentrations and pharmacological effects of co-administered drugs. [1] |
| Molecular Formula |
C27H34O11
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|---|---|
| Molecular Weight |
534.5523
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| Exact Mass |
534.21
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| Elemental Analysis |
C, 60.67; H, 6.41; O, 32.92
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| CAS # |
487-41-2
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
730.4±60.0 °C at 760 mmHg
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| Flash Point |
395.5±32.9 °C
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| Vapour Pressure |
0.0±2.5 mmHg at 25°C
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| Index of Refraction |
1.596
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| LogP |
0.08
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| SMILES |
O1C([H])([H])[C@]2([H])[C@]([H])(C3C([H])=C([H])C(=C(C=3[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[C@]2([H])[C@@]1([H])C1C([H])=C([H])C(=C([H])C=1OC([H])([H])[H])O[C@@]1([H])C([H])(C([H])([C@@]([H])(C([H])(C([H])([H])O[H])O1)O[H])O[H])O[H]
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| InChi Key |
KFFCKOBAHMGTMW-LGQRSHAYSA-N
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| InChi Code |
InChI=1S/C27H34O11/c1-32-17-6-4-13(8-19(17)33-2)25-15-11-36-26(16(15)12-35-25)14-5-7-18(20(9-14)34-3)37-27-24(31)23(30)22(29)21(10-28)38-27/h4-9,15-16,21-31H,10-12H2,1-3H3/t15-,16-,21+,22+,23-,24+,25-,26+,27+/m0/s1
|
| Chemical Name |
(2S,3R,4S,5S,6R)-2-[4-[(3R,3aR,6S,6aR)-3-(3,4-dimethoxyphenyl)-1,3,3a,4,6,6a-hexahydrofuro[3,4-c]furan-6-yl]-2-methoxyphenoxy]-6-(hydroxymethyl)oxane-3,4,5-triol
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| Synonyms |
Phillyrin
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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 : ~250 mg/mL (~467.68 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.17 mg/mL (4.06 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 21.7 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: ≥ 2.17 mg/mL (4.06 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 21.7 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: ≥ 2.17 mg/mL (4.06 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 | 1.8707 mL | 9.3537 mL | 18.7073 mL | |
| 5 mM | 0.3741 mL | 1.8707 mL | 3.7415 mL | |
| 10 mM | 0.1871 mL | 0.9354 mL | 1.8707 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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