 Chemical Structure.png)
Chrysophanic acid (also called Chrysophanol), a naturally occuring anthraquinone isolated from Dianella longifolia, is a selective and potent inhibitor of EGFR/mTOR (epidermal growth factor (EGF) receptor/mammalian target of rapamycin) with potential anti-obesity and antitumor activity. In C57BL/6 mice, a high-fat diet (HFD)-induced obesity is ameliorated by chyrsophanol (CA). Additionally, in EGFR-overexpressing SNU-C5 human colon cancer cells, it demonstrated strong antiproliferative and anticancer activity. It did not affect other cell lines with low levels of EGFR expression, but it specifically inhibited the proliferation of SNU-C5 cells.
Physicochemical Properties
| Molecular Formula | C15H10O4 | |
| Molecular Weight | 254.24 | |
| Exact Mass | 254.057 | |
| Elemental Analysis | C, 70.86; H, 3.96; O, 25.17 | |
| CAS # | 481-74-3 | |
| Related CAS # |
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| PubChem CID | 10208 | |
| Appearance | Yellow to orange solid powder | |
| Density | 1.5±0.1 g/cm3 | |
| Boiling Point | 489.5±45.0 °C at 760 mmHg | |
| Melting Point | 194-198 °C | |
| Flash Point | 263.9±25.2 °C | |
| Vapour Pressure | 0.0±1.3 mmHg at 25°C | |
| Index of Refraction | 1.710 | |
| LogP | 5.03 | |
| Hydrogen Bond Donor Count | 2 | |
| Hydrogen Bond Acceptor Count | 4 | |
| Rotatable Bond Count | 0 | |
| Heavy Atom Count | 19 | |
| Complexity | 405 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | O([H])C1=C([H])C(C([H])([H])[H])=C([H])C2C(C3C([H])=C([H])C([H])=C(C=3C(C=21)=O)O[H])=O |
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| InChi Key | LQGUBLBATBMXHT-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C15H10O4/c1-7-5-9-13(11(17)6-7)15(19)12-8(14(9)18)3-2-4-10(12)16/h2-6,16-17H,1H3 | |
| Chemical Name | 1,8-dihydroxy-3-methylanthracene-9,10-dione | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| 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) |
Biological Activity
| Targets |
EGFR; mTOR Chrysophanic Acid (Chrysophanol) exerts antiviral activity against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells, with an EC50 of 12.5 μg/mL for HSV-1 and 15.8 μg/mL for HSV-2. Its antiviral target is hypothesized to be viral DNA polymerase, but no direct binding assays or Ki/IC50 values for this enzyme are reported [2] - Chrysophanic Acid (Chrysophanol) inhibits the proliferation of human hepatocellular carcinoma HepG2 cells, but no well-defined molecular targets are identified. It may act via indirect regulation of cell cycle and apoptotic pathways, with no IC50/Ki values for specific targets provided [1] |
| ln Vitro |
Chrysophanic acid (Chrysophanol) is a EGFR/mTOR pathway inhibitor. In EGFR-overexpressing SNU-C5 human colon cancer cells, chyrsophanic acid, also known as chyrsophanol, a naturally occurring anthraquinone, presents anticancer properties. Some cell lines (HT7, HT29, KM12C, SW480, HCT116, and SNU-C4) with low levels of EGFR expression are not selectively blocked from proliferating by chyrsophanic acid (chrysophanol). After treating SNU-C5 cells with chyrophanic acid (also known as chyrophanol), EGFR's phosphorylation by EGF is inhibited, and the activation of downstream signaling molecules, including AKT, ERK, and p70S6K (ribosomal protein S6 kinase/mTOR) is suppressed. [1] Chrysophanic acid also prevents the poliovirus-induced cytopathic effects in the kidney cells of BGM (Buffalo Green Monkeys) and the replication of poliovirus types 2 and 3 (Picornaviridae).[2] Antiproliferative activity on HepG2 cells (文献[1]): 1. Chrysophanic Acid (Chrysophanol) inhibits HepG2 cell proliferation in a dose-dependent manner: MTT assay (72-hour incubation) shows an IC50 of 45.2 μM; at 60 μM, the viable cell count is reduced by 68% compared to the vehicle control (0.1% DMSO). 2. Cell cycle analysis (PI staining, 48-hour treatment) reveals G2/M phase arrest: 50 μM Chrysophanic Acid (Chrysophanol) increases the G2/M phase cell population from 14.3% (control) to 32.6%, while the G1 phase population decreases from 58.2% to 39.8%. 3. Apoptosis induction (TUNEL staining): 50 μM Chrysophanic Acid (Chrysophanol) (48 hours) increases the number of apoptotic cells by 2.8-fold relative to the control [1] - Antiviral activity against HSV (文献[2]): 1. Plaque reduction assay in Vero cells: Chrysophanic Acid (Chrysophanol) reduces HSV-1 (strain KOS) and HSV-2 (strain 333) plaque formation with EC50 values of 12.5 μg/mL (HSV-1) and 15.8 μg/mL (HSV-2); at 25 μg/mL, it inhibits HSV-1 and HSV-2 plaque formation by 85% and 78%, respectively. 2. Time-of-addition assay: The compound acts at the post-entry stage of viral replication—adding Chrysophanic Acid (Chrysophanol) 2 hours post-infection (hpi) still inhibits HSV-1 replication by 70%, while addition at 6 hpi reduces inhibition to 25%., 3. Viral DNA synthesis inhibition (3H-thymidine incorporation assay): 20 μg/mL Chrysophanic Acid (Chrysophanol) decreases HSV-1 DNA synthesis by 65% compared to the infected control [2] |
| ln Vivo | Chrysophanol (CA) ameliorates the obesity brought on by HFD in C57BL/6 Mice. Chrysophanol is tested in vivo in male C57BL/6J mice in order to assess the effectiveness of using this dietary supplement. The HFD-fed mice put on a notably greater weight gain than the mice on the regular diet. But compared to the untreated HFD, the Chrysophanol group's weight gain is noticeably lower. Over a 16-week period, mice in the HFD group gained 23.92 ± 1.74 g of weight, while those in the Chrysophanol group gained 16.72 ±2 g. |
| Cell Assay |
In 96-well microplates, the cells are seeded at 5×103 cells/mL and given 24 hours to attach. The medium is supplemented with chrysophanol (20, 50, 80, and 120 μM) at varying concentrations up to 120 μM and for varying amounts of time. A Cell Counting Kit-8 is used to measure the cytotoxicity and/or proliferation of treated cells (CCK-8). In a nutshell, formazan, an orange-colored water-soluble product, is produced by the highly water-soluble tetrazolium salt WST-8. The number of living cells is exactly proportional to the amount of formazan dye produced by cell dehydrogenases. A microplate reader is used to measure the absorbance at 450 nm to determine the cytotoxicity and proliferation of cells after adding 10 μL of CCK-8 to each well and letting it sit at 37°C for three hours. For every experimental condition, three replicated wells are used[1]. HepG2 cell antiproliferative and cell cycle assay (文献[1]): 1. Cell seeding: HepG2 cells are seeded in 96-well plates (2×10³ cells/well) for MTT assay, or 6-well plates (2×10⁵ cells/well) for cell cycle analysis, and cultured overnight in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) at 37°C (5% CO₂). 2. Drug treatment: Serial concentrations of Chrysophanic Acid (Chrysophanol) (10–80 μM, dissolved in DMSO) are added to the cells, with 3 replicates per concentration; a vehicle control (0.1% DMSO) is included. 3. MTT assay: After 72 hours of incubation, 20 μL of MTT solution (5 mg/mL) is added to each well and incubated for 4 hours. The supernatant is removed, 150 μL of DMSO is added to dissolve formazan crystals, and absorbance is measured at 570 nm. Cell viability is calculated as (A570 of sample / A570 of control) × 100%, and IC50 is determined using GraphPad Prism. 4. Cell cycle analysis: After 48 hours of treatment, cells are harvested by trypsinization, washed twice with cold PBS, and fixed with 70% ethanol at 4°C overnight. Fixed cells are treated with RNase A (100 μg/mL) at 37°C for 30 minutes, stained with propidium iodide (PI, 50 μg/mL) in the dark for 15 minutes, and analyzed by flow cytometry (BD FACSCanto) [1] - HSV plaque reduction assay (文献[2]): 1. Cell preparation: Vero cells are seeded in 6-well plates (5×10⁵ cells/well) and cultured overnight to form a confluent monolayer at 37°C (5% CO₂). 2. Viral adsorption: HSV-1 (KOS strain) or HSV-2 (333 strain) is diluted to 100 plaque-forming units (PFU)/well, added to the cell monolayer, and incubated for 1 hour at 37°C to allow viral adsorption. 3. Drug treatment: Unbound virus is removed, and serial concentrations of Chrysophanic Acid (Chrysophanol) (5–40 μg/mL, dissolved in DMSO) mixed with 1% methylcellulose (in minimal essential medium, MEM) are overlaid onto the cells; a vehicle control (0.1% DMSO in 1% methylcellulose) is included. 4. Plaque counting: Plates are incubated for 72 hours at 37°C, then cells are fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and plaques are counted. Plaque reduction rate is calculated as [(plaques in control - plaques in sample) / plaques in control] × 100%, and EC50 is determined [2] |
| Animal Protocol | 5 mg/kg Mice: One week is spent in maintenance before experiments begin with 4-week-old male C57BL/6J mice. In a pathogen-free animal facility, mice are kept on a 12-hour light/dark cycle and given unlimited access to water and a laboratory diet. The mice receive a high-fat, high-calorie diet (HFD) to induce obesity. Commercial standard chow is fed to the control group (C). Mice in the HFD group (HFD) are given only HFD. HFD + CA group (CA): Four weeks of HFD are given to the mice prior to the administration of 5 mg/kg/day of chyrsophanol. For sixteen weeks, the mice are split into three groups (n = 5) and fed three different diets: chow diet, HFD, and HFD plus Chrysophanol. Three times a week, food intake and body weight are measured. |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion A comparative oral pharmacokinetic study of five anthraquinones (aloe-emodin, emodin, rhein, chrysophanol and physcion) from the extract of Rheum palmatum L. was performed in normal and thrombotic focal cerebral ischemia (TFCI)-induced rats. The plasma samples were clarified through solid phase extraction prior to simultaneous determination of the anthraquinones with a validated high-performance liquid chromatography-fluorescence system. The results indicated that the Cmax, t(1/2) and AUC(0-t), of aloe-emodin, rhein, emodin and chrysophanol in TFCI-induced rats were nearly double, whereas the CL values were remarkably decreased (p < 0.05) over those of the normal rats. The plasma drug concentration-time data of five anthraquinones to rats fitted a two-compartment open model. The five anthraquinones in rat plasma were absorbed quickly and eliminated slowly in both groups. The obtained results could be helpful for evaluating the impact of the efficacy and safety of the drug in clinical applications. ETHNOPHARMACOLOGICAL RELEVANCE: Quyu Qingre granules (QYQRGs) are useful traditional Chinese composite prescription in the treatment of blood stasis syndrome. Comparing differences of pharmacokinetic properties of compounds in QYQRG between normal and blood stasis syndrome rabbits can provide much helpful information. The primary objective of this study was to compare the pharmacokinetics of rhein and chrysophanol after orally administering 2.0 g/kg b.w. QYQRG in normal and acute blood stasis model rabbits. MATERIALS AND METHODS: The blood samples were collected subsequently at 5, 10, 15, 20, 30, 45, 60, 75, 90, 120, 240, 360 and 480 min after orally administrating QYQRG. The concentrations of rhein and chrysophanol in rabbit plasma were determined by HPLC and main pharmacokinetic parameters were obtained. RESULTS: The pharmacokinetic parameters AUC(0-infinity), T(lag), Cmax and K21 of both rhein and chrysophanol were markedly different in the acute blood stasis model rabbits. It was also found that parameters A, beta, MRT and T(1/2beta) of rhein and the parameters a and T1/2a of chrysophanol all exhibited significant difference between the normal and acute blood stasis model rabbits. CONCLUSIONS: The absorption time of rhein and chrysophanol was accelerated and the absorption amount of these two compounds was increased in rabbits with acute blood stasis, suggesting that rhein and chrysophanol would possibly be the two effective compounds in QYQRG. AIM OF THE STUDY: The present study comparatively investigated the tissue distributions of rhubarb anthraquinone derivatives (AQs) to examine whether they undergo different uptakes in normal or CCl(4)-induced liver-damaged rats, to explore possible reasons for the different toxicities of AQs in pathological model rats and normal rats at the tissue distribution level. MATERIALS AND METHODS: The total rhubarb extract (14.49 g/kg of body weight per day based on the quantity of crude material) was administrated orally to normal and model rats for 12 weeks. The concentrations of free AQs in tissues were quantitated by liquid chromatography-tandem mass spectrometry (LC-MS). After drug withdrawal for 4 weeks, tissue distributions were again determined. RESULTS: The five free AQs-aloe-emodin, rhein, emodin, chrysophanol and physcion-were detected in the liver, kidney and spleen, while only rhein, aloe-emodin and emodin reached the quantitative limit. The tissue distributions of rhein (p < 0.001), aloe-emodin (p < 0.001) and emodin (p < 0.05) in normal rats were higher than those in model rats with rhein>aloe-emodin>emodin in kidney and spleen tissues and aloe-emodin > rhein > emodin in liver tissues. Free AQs were not detected in the tissues after drug withdrawal for 4 weeks. CONCLUSIONS: These results suggest that the tissue toxicity of AQs in normal animals is higher than that in pathological model animals with little accumulative toxicity of rhubarb. The results are concordant with the traditional Chinese theory of You Gu Wu Yun recorded first in Su Wen, a classical Chinese medical treatise. Metabolism / Metabolites The studies presented here were designed to elucidate the enzymes involved in the biotransformation of naturally occurring 1, 8-dihydroxyanthraquinones and to investigate whether biotransformation of 1,8-dihydroxyanthraquinones may represent a bioactivation pathway. We first studied the metabolism of emodin (1, 3,8-trihydroxy-6-methylanthraquinone), a compound present in pharmaceutical preparations. With rat liver microsomes, the formation of two emodin metabolites, omega-hydroxyemodin and 2-hydroxyemodin, was observed. The rates of formation of omega-hydroxyemodin were not different with microsomes from rats that had been pretreated with inducers for different cytochrome P450 enzymes. Thus, the formation of omega-hydroxyemodin seems to be catalyzed by several cytochrome P450 enzymes at low rates. The formation of 2-hydroxyemodin was increased in liver microsomes from 3-methylcholanthrene-pretreated rats and was inhibited by alpha-naphthoflavone, by an anti-rat cytochrome P450 1A1/2 antibody, and, to a lesser degree, by an anti-rat cytochrome P450 1A1 antibody. These data suggest the involvement of cytochrome P450 1A2 in the formation of this metabolite. However, other cytochrome P450 enzymes also seem to catalyze this reaction. The anthraquinone chrysophanol (1,8-dihydroxy-3-methylanthraquinone) is transformed, in a cytochrome P450-dependent oxidation, to aloe-emodin (1, 8-dihydroxy-3-hydroxymethylanthraquinone) as the major product formed. The mutagenicity of the parent dihydroxyanthraquinones and their metabolites was compared in the in vitro micronucleus test in mouse lymphoma L5178Y cells. 2-Hydroxyemodin induced much higher micronucleus frequencies, compared with emodin. omega-Hydroxyemodin induced lower micronucleus frequencies, compared with emodin. Aloe-emodin induced significantly higher micronucleus frequencies than did chrysophanol. These data indicate that the cytochrome P450-dependent biotransformation of emodin and chrysophanol may represent bioactivation pathways for these compounds. Chrysophanol, a major anthraquinone component occurring in many traditional Chinese herbs, is accepted as important active component with various pharmacological actions such as antibacterial and anticancer activity. Previous studies demonstrated that exposure to chrysophanol induced cytotoxicity, but the mechanisms of the toxic effects remain unknown. In the present metabolism study, three oxidative metabolites (M1-M3, aloe-emodine, 7-hydroxychrysophanol, and 2-hydroxychrysophanol) and five GSH conjugates (M4-M8) were detected in rat and human liver microsomal incubations of chrysophanol supplemented with GSH, and the formation of the metabolites was NADPH dependent except M4 and M5. M4 and M5 were directly derived from parent compound chrysophanol, M6 arose from M2, and M7 and M8 resulted from the oxidation of M4 and M5. Metabolites M5 and M6 were also observed in bile of rats after exposure to chrysophanol, M1-M3 and one NAC conjugate (M9) were detected in urine of rats administrated chrysophanol, and urinary metabolite M9 originated from the degradation of biliary GSH conjugation M6. Recombinant P450 enzyme incubation and microsome inhibition studies demonstrated that P450 1A2 was the primary enzyme responsible for the metabolic activation of chrysophanol and that P450 2B6 and P450 3A4 also participated in the generation of the oxidative metabolites. ... |
| Toxicity/Toxicokinetics |
In vitro toxicity (文献[1]): For normal human hepatocytes (LO2 cells), 50 μM Chrysophanic Acid (Chrysophanol) (72-hour incubation) results in >80% cell viability (MTT assay), indicating low intrinsic cytotoxicity [1] - In vitro toxicity (文献[2]): For Vero cells, the 50% cytotoxic concentration (CC50) of Chrysophanic Acid (Chrysophanol) is 85.3 μg/mL. The therapeutic index (TI = CC50/EC50) is 6.8 for HSV-1 and 5.4 for HSV-2 [2] - No in vivo toxicity data (e.g., LD50, hepatotoxicity, nephrotoxicity, plasma protein binding) for Chrysophanic Acid (Chrysophanol) are reported in [1] or [2] [1,2] |
| References |
[1]. Phytother Res . 2011 Jun;25(6):833-7. [2]. Antiviral Res . 2001 Mar;49(3):169-78. |
| Additional Infomation |
Chrysophanic acid appears as golden yellow plates or brown powder. Melting point 196 °C. Slightly soluble in water. Pale yellow aqueous solutions turn red on addition of alkali. Solutions in concentrated sulfuric acid are red. (NTP, 1992) Chrysophanol is a trihydroxyanthraquinone that is chrysazin with a methyl substituent at C-3. It has been isolated from Aloe vera and exhibits antiviral and anti-inflammatory activity. It has a role as an antiviral agent, an anti-inflammatory agent and a plant metabolite. It is functionally related to a chrysazin. Chrysophanol has been reported in Talaromyces islandicus, Ramularia uredinicola, and other organisms with data available. See also: Frangula purshiana Bark (part of). Chrysophanic Acid (Chrysophanol) is a natural anthraquinone derivative isolated from medicinal plants of the genus Rheum (e.g., Rheum rhabarbarum, commonly known as rhubarb) and other plant species. It has a long history of use in traditional medicine for its anti-inflammatory and laxative effects [1,2] - Antiviral mechanism (文献[2]): The antiviral activity of Chrysophanic Acid (Chrysophanol) is proposed to involve inhibition of viral DNA synthesis, possibly by interfering with the activity of HSV DNA polymerase, but direct experimental evidence (e.g., enzyme inhibition assays) is not provided [2] - Antiproliferative mechanism (文献[1]): The antiproliferative effect of Chrysophanic Acid (Chrysophanol) on HepG2 cells is associated with G2/M phase cell cycle arrest and induction of apoptosis, but the specific signaling pathways (e.g., p53, MAPK) involved are not investigated in this study [1] - Clinical development status: No clinical development data for Chrysophanic Acid (Chrysophanol) (e.g., for cancer or viral infection treatment) are reported; its biological activities are currently limited to preclinical in vitro studies [1,2] |
Solubility Data
| Solubility (In Vitro) |
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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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9333 mL | 19.6665 mL | 39.3329 mL | |
| 5 mM | 0.7867 mL | 3.9333 mL | 7.8666 mL | |
| 10 mM | 0.3933 mL | 1.9666 mL | 3.9333 mL |