naphthoquinones and acetylshikonin

Acetylshikonin, a naphthoquinone derivative, is mainly extracted from some species of the family Boraginaceae, such as. This review provides a comprehensive summary of the pharmacology, toxicity, and pharmacokinetics of acetylshikonin focussing on its mechanisms on the basis of currently available literature.. The information of acetylshikonin from 1977 to 2020 was collected using major databases including Elsevier, Scholar, PubMed, Springer, Web of Science, and CNKI. Acetylshikonin, pharmacology, toxicity, pharmacokinetics, and naphthoquinone derivative were used as key words.. According to emerging evidence, acetylshikonin exerts a wide spectrum of pharmacological effects such as anticancer, anti-inflammatory, lipid-regulatory, antidiabetic, antibacterial, antifungal, antioxidative, neuroprotective, and antiviral properties. However, only a few studies have reported the adverse effects of acetylshikonin, with respect to reproductive toxicity and genotoxicity. Pharmacokinetic studies demonstrate that acetylshikonin is associated with a wide distribution and poor absorption.. Although experimental data supports the beneficial effects of this compound, acetylshikonin cannot be considered as a therapy drug without further investigations, especially, on the toxicity and pharmacokinetics.

Topics: Animals; Anthraquinones; Boraginaceae; Drugs, Chinese Herbal; Humans; Mice; Naphthoquinones; Plant Extracts; Rats

Shikonin (SH) is used as a red pigment for food coloring and cosmetics, and has cytotoxic activity towards cancer cells. However, due to strong toxicity SH has limited potential as an anticancer drug. Acetylshikonin (ASH) is one of the SH derivatives with promising anticancer potential. In present study, we attempted to evaluate and compare the cytotoxicity of SH and ASH towards a normal cell line (V79) and in addition to evaluate their antigenotoxic activity. The evaluation was made with the use of the set of cytotoxicity assays with V79 line and the micronucleus test

Topics: Animals; Anthraquinones; Cell Line; Cricetulus; Cyclophosphamide; DNA Damage; Ethyl Methanesulfonate; Fluoroquinolones; Micronucleus Tests; Naphthoquinones

In the present study, five root extracts of Onosma visianii Clem were investigated for their in vitro cytotoxic activity. On the basis of HPLC-PDA analysis, these extracts have proved to be a rich source of naphthoquinones as natural colourants for food and cosmetic industry. All investigated root extracts contain acetylshikonin, isobutyrylshikonin and α-methylbutyrylshikonin as major compounds. As the most abundant source of active compounds for antitumour therapy, acetone, chloroform and ethyl acetate extracts showed strong cytotoxic activity towards HCT-116 and MDA-MB-231 cancer cell lines. Also, these extracts induced apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

Topics: Anthraquinones; Antineoplastic Agents, Phytogenic; Apoptosis; Boraginaceae; Cell Cycle Checkpoints; Cell Line, Tumor; Humans; Molecular Structure; Naphthoquinones; Phytochemicals; Plant Extracts; Plant Roots

Cytotoxic activities of acetylshikonin and acetoxyisovalerylshikonin alone and in combination with chemotherapeutic agents against parental and drug resistant cell lines were determined using the MTT assay. Effects of Shikonin derivatives on BCRP, MDR1 and MRP transcript and protein levels were relatively measured. Finally, accumulation and efflux kinetics were conducted. The results revealed cell- and concentration-dependency of the cell cytotoxicity. Acetylshikonin and acetoxyisovalerylshikonin transiently made the mRNA ocean turbulent, but FACS analyses using fluorescent-labeled antibodies showed no significant change in the MDR-protein levels. Functional kinetics revealed significant block of MDR1, BCRP and MRP transporter in the presence of shikonin derivatives. Maximum accumulation fold changes was quantified to be 4.4 and consequently, acetoxyisovalerylshikonin pretreated EPG85.257RDB cells was chemosensitized to daunorubicin tension 3.1-fold. Although, the MDR blockage was reported to follow time- and cell-dependent patterns, MDR1, BCRP and MRP2 responses to the shikonins are concentration-independent. These data suggest uncompetitive transporter blockage behavior of these agents. The results indicated that shikonin derivatives stimulate uptake and reduce efflux of chemotherapeutic agents in the malignant cancer cells, suggesting that chemotherapy in combination with shikonin compounds may be beneficial to cancer cells that overexpress multidrug resistance transporters.

Topics: Anthraquinones; Antineoplastic Agents; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP Binding Cassette Transporter, Subfamily G, Member 2; Cell Line, Tumor; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Therapy, Combination; Humans; Multidrug Resistance-Associated Proteins; Naphthoquinones; Neoplasm Proteins; Neoplasms; Phenotype

Overexpression and mutation of the epidermal growth factor receptor (EGFR) gene play a causal role in tumorigenesis and resistance to treatment of glioblastoma (GBM). EGFR inhibitors such as erlotinib are currently used for the treatment of GBM; however, their efficacy has been limited due to drug resistance. New treatment strategies are therefore urgently needed. Shikonin, a natural naphthoquinone, induces both apoptosis and necroptosis in human glioma cells, but the effectiveness of erlotinib-shikonin combination treatment as well as the underlying molecular mechanisms is unknown yet. In this study, we investigated erlotinib in combination with shikonin and 14 shikonin derivatives in parental U87MG and transfected U87MG.ΔEGFR GBM cells. Most of the shikonin derivatives revealed strong cytotoxicity. Shikonin together with five other derivatives, namely deoxyshikonin, isobutyrylshikonin, acetylshikonin, β,β-dimethylacrylshikonin and acetylalkannin showed synergistic cytotoxicity toward U87MG.ΔEGFR in combination with erlotinib. Moreover, the combined cytotoxic effect of shikonin and erlotinib was further confirmed with another three EGFR-expressing cell lines, BS153, A431 and DK-MG. Shikonin not only dose-dependently inhibited EGFR phosphorylation and decreased phosphorylation of EGFR downstream molecules, including AKT, P44/42MAPK and PLCγ1, but also together with erlotinib synergistically inhibited ΔEGFR phosphorylation in U87MG.ΔEGFR cells as determined by Loewe additivity and Bliss independence drug interaction models. These results suggest that the combination of erlotinib with shikonin or its derivatives might be a potential strategy to overcome drug resistance to erlotinib.

Topics: Anthraquinones; Antineoplastic Combined Chemotherapy Protocols; Cell Line, Tumor; Drug Synergism; ErbB Receptors; Erlotinib Hydrochloride; Glioblastoma; Humans; Mitogen-Activated Protein Kinases; Naphthoquinones; Phosphorylation; Proto-Oncogene Proteins c-akt; Quinazolines; Signal Transduction

The aim of this study was to evaluate the antigenotoxic and antioxidant potential of shikonin (SH), acetylshikonin (ACS) and Arnebia euchroma callus extract (EXT). The antigenotoxic activity was investigated by the umu-test as the inhibition of the SOS system induction caused by genotoxic chemical agents - 4-nitroquinoline oxide and 2-aminoanthracene. Moreover the ability of SH, ACS and EXT to prevent photogenotoxicity triggered by chlorpromazine under UVA irradiation was measured. The cytotoxicity of EXT toward V79 Chinese hamster cell line was additionally assessed. Shikonin and acetylshikonin had no effect on 4-NQO induced genotoxicity whereas EXT demonstrated an unclear effect. The protection against 2AA induced genotoxicity was observed for all tested substances. The highest protection was demonstrated for EXT with inhibition of 66%. SH and ACS reduced 2AA genotoxicity with inhibition of about 60%. Under UVA the strongest and dose-dependent activity was observed for EXT. Acetylshikonin was a weak anti-photogenotoxin whereas shikonin had no clear effect. EXT was highly cytotoxic toward the V79 cell line - the cells' morphology was affected seriously and apoptosis was impacted. The antioxidant activity of SH, ACS and EXT was studied by means of electron paramagnetic resonance spectroscopy using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. All three samples exhibited radical scavenging properties.

Topics: 4-Nitroquinoline-1-oxide; Animals; Anthracenes; Anthraquinones; Antioxidants; Boraginaceae; Cell Line; Chlorpromazine; Cricetinae; Cricetulus; Electron Spin Resonance Spectroscopy; Male; Mutagenicity Tests; Naphthoquinones; Plant Extracts; Rats; Rats, Sprague-Dawley

Topics: Anthraquinones; Drugs, Chinese Herbal; Ions; Naphthoquinones; Pentanoic Acids; Spectrometry, Mass, Electrospray Ionization

Aikete injection is composed of acetylshikonin and β,β-dimethylacrylshikonin, which have been reported to have anti-tumor effects on a wide range of cancer cell lines. However, little is known about the effects of the combination of the two components on cancer cells.. To investigate the anti-proliferation activity of Aikete injection on human hepatocellular carcinoma (HCC) cells and its mechanism.. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and growth curve assay were used to determine the inhibitory effect of Aikete injection on the proliferation of SMMC-7721 cells. Giemsa staining, Hoechst 33258 staining and flow cytometry were used to assess cell apoptosis. Expression of Bcl-2 and Bax was analyzed by reverse transcription-polymerase chain reaction and flow cytometry. H22 bearing mice were also used to determine the anti-tumor effect of Aikete injection in vivo.. Aikete injection inhibited the proliferation of SMMC-7721 cells in both a dose- and time-dependent manner in vitro. The characteristics of apoptosis were observed in Aikete injection groups by Hoechst 33258 and Giemsa staining. In addition, Aikete injection induced cell cycle arrest at G2/M phase and downregulated the Bcl-2 expression and the ratio of Bcl-2/Bax in SMMC-7721 cells. The experiment in vivo showed that Aikete injection significantly inhibited the growth of H22 carcinoma, with an inhibitory rate of 34.37-57.99%.. The results demonstrated that Aikete injection suppressed the growth of HCC cells in vitro and in vivo by inducing cell apoptosis.

Topics: Animals; Anthraquinones; Antineoplastic Agents, Phytogenic; Apoptosis; Carcinoma, Hepatocellular; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Genes, bcl-2; Humans; Injections; Liver Neoplasms; Male; Mice; Naphthoquinones

A simple, precise, and rapid high-performance thin-layer chromatographic (HPTLC) method for the simultaneous quantification of pharmacologically important naphthoquinone shikonin (1) together with its derivatives acetylshikonin (2), and beta-acetoxyisovalerylshikonin (3) in four species of genus Arnebia (A. euchroma, A. guttata, A. benthamii, and A. hispidissima) from the Indian subcontinent has been developed. In addition, the effect of solvents with varying polarity (hexane, chloroform, ethyl acetate, and methanol) for the extraction of these compounds was studied. HPTLC was performed on precoated RP-18 F(254S )TLC plates. For achieving good separation, mobile phase consisting of ACN/methanol/5% formic acid in water (40:02:08 v/v/v) was used. The densitometric determination of shikonin derivatives was carried out at 520 nm in reflection/absorption mode. The method was validated in terms of linearity, accuracy, precision, robustness, and specificity. The calibration curves were linear in the range of 100-600 ng for shikonin and acetylshikonin, and 100-1800 ng for beta-acetoxyisovalerylshikonin. Lower LOD obtained for compounds 1-3 were 18, 15, and 12 ng, respectively, while the LOQ obtained were 60, 45, and 40 ng, respectively.

Topics: Anthraquinones; Boraginaceae; Chromatography, Thin Layer; Densitometry; Naphthoquinones; Species Specificity; Ultrasonics

Lithospermum erythrorhizon has been used for treatment of inflammatory diseases and cancer as a folk remedy. Based on the evidences that anti-inflammatory agents frequently exert antiangiogenic activity, thus we examined comparatively the antiangiogenic activities of three naphthoquinone derivatives (shikonin, acetylshikonin, and isobutyroylshikonin) isolated from the plant. Three derivatives exhibited weak cytotoxicity against human umbilical vein endothelial cells (HUVECs) with IC50 of over 20 microM. Shikonin had more specific inhibitory effects on proliferation and vascular endothelial growth factor (VEGF) production by VEGF compared with different derivatives. All of derivatives significantly suppressed the migration of VEGF treated HUVECs at different optimal concentrations. Also, shikonin and acetylshikonin significantly disrupted VEGF-induced tube formation. Furthermore, three derivatives effectively downregulated the expression of urokinase-type plasminogen activator (uPA), but not its receptor uPAR. Additionally, shikonin significantly inhibited tumor growth in LLC-bearing mice, whereas its derivatives had relatively mild effects. Taken together, our findings suggest that shikonin and its derivatives exhibit the antiangiogenic and antitumorigenic effects by suppressing proliferation and angiogenic factors.

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents, Non-Steroidal; Carcinoma, Lewis Lung; Cell Movement; Cells, Cultured; Depression, Chemical; Dose-Response Relationship, Drug; Down-Regulation; Endothelial Cells; Female; Humans; Lithospermum; Mice; Naphthoquinones; Neovascularization, Pathologic; Urokinase-Type Plasminogen Activator; Vascular Endothelial Growth Factor A

Shikonin and its derivatives are important red colored naphthoquinone pigments found in a large number of Arnebia species, including A. euchroma, that are responsible for the various pharmacological activities exhibited by the plant. The precise separation of each naphthoquinone is essential for total quality evaluation and bioactivity analysis of herbal formulations of A. euchroma. Furthermore, the overexploitation of this useful plant has resulted in species becoming endangered. With this in mind, a simple and rapid preparative scale HPLC method with single compound recovery for the isolation and purification of two shikonin derivatives (i. e. acetylshikonin, beta-acetoxyisovalerylshikonin) from cell suspension cultures of A. euchroma is presented. The compounds were separated on a C(18) column within 10 min using acetonitrile/methanol (95:5) as mobile phase in isocratic mode. The isolated compounds were found to be more than 98% pure. The LOD for acetylshikonin and beta-acetoxyisovalerylshikonin was estimated at 0.063 and 0.146 mug/mL, respectively, while the LOQ was found to be 0.209 and 0.487 mug/mL, respectively. The recoveries accomplished for both the shikonin derivatives were in the range of 94.7-96.8%. The repeatability, expressed as %RSD, of acetylshikonin and beta-acetoxyisovalerylshikonin was found to be 1.74 and 1.27, respectively.

Topics: Anthraquinones; Boraginaceae; Cells, Cultured; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Naphthoquinones; Spectrophotometry, Ultraviolet

The root extracts of Onosma leptanhtha were evaluated for their anti-iflammatory and cytotoxic activities. The cyclohexane extract, which appeared as the most active in both assays, has been further subjected to bioassay-directed fractionation to afford the naphthazarine derivatives: beta,beta-dimethylacrylshikonin (1), isovalerylshikonin (2) and acetylshikonin (3). The evaluation of the anti-inflammatory activity was performed on carrageenan-induced rat paw edema test. All the tested compounds proved to be active, while compound 3 showed the best anti-inflammatory effect. In addition, the cytotoxic activity of the extracts and isolated compounds, was also assayed against L1210 murine lymphoblastic leukemia cell line, and human fibrosarcoma HT-1080 cells. Compound 1 exhibited remarkable cytotoxic activity (390 nM for L1210 cells), which is superior to that of shikonin, which was used as control.

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents; Antineoplastic Agents; Biological Assay; Boraginaceae; Carrageenan; Cell Line, Tumor; Cyclohexanes; Edema; Humans; Indomethacin; Inhibitory Concentration 50; Male; Naphthoquinones; Plant Extracts; Plant Roots; Rats; Rats, Wistar

A high-performance liquid chromatography (HPLC) analysis system based on a water-acetonitrile gradient program was established for simultaneous quantification of shikimate-derived secondary metabolites in cultured cells of Lithospermum erythrorhizon. The cells cultured in pigment production medium (M-9) are capable of producing five highly hydrophilic compounds such as p-hydroxybenzoic acid-O-glucoside and lithospermic acid B, as well as eleven lipophilic compounds including echinofuran B and acetylshikonin. In addition to the wide polarities of those compounds, many of them are unstable under light, dryness, oxygen and heating. Thus, a new extraction procedure for all these compounds was also established by use of ultrasonication under ice-water chilling with MeOH as the solvent. This procedure was applied to the quantitative analyses of these compounds in cell cultures and hairy root cultures of Lithospermum, and in the intact plants as well.

Topics: Anthraquinones; Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents, Phytogenic; Benzofurans; Cells, Cultured; Chromatography, High Pressure Liquid; Cold Temperature; Depsides; Furans; Glucosides; Naphthoquinones; Parabens; Plant Roots; Plants, Medicinal; Shikimic Acid; Solvents; Sonication

Acetylshikonin, teracrylshikonin, beta,beta-dimethylacrylshikonin and shikonin, isolated from Arnebia euchroma, inhibited collagen (10 micrograms/ml)-induced aggregation of washed rabbit platelets in a concentration-dependent manner with IC50 values of 2.1 +/- 0.2, 2.8 +/- 0.3, 4.2 +/- 0.5 and 10.7 +/- 0.7 microM, respectively. Acetylshikonin also inhibited the aggregation and ATP release of washed rabbit platelets induced by arachidonic acid (AA, 100 microM), U46619 (1 microM), platelet-activating factor (PAF, 3.6 nM) and thrombin (0.1 U/ml) in a concentration-dependent manner. The IC50 values of acetylshikonin on the inhibition of these four agonists-induced platelet aggregation were 3.1 +/- 0.4, 2.2 +/- 0.2, 8.0 +/- 0.6 and 12.7 +/- 1.0 microM, respectively. The thromboxane B2 formation caused by collagen, PAF and thrombin was inhibited by acetylshikonin, while formations of thromboxane B2 and prostaglandin D2 caused by AA were not inhibited. Acetylshikonin did not inhibit cyclooxygenase activity since it did not attenuate prostaglandin E2 formation after incubation of sheep vesicular gland microsomes with AA. Acetylshikonin suppressed both the rise of intracellular Ca2+ concentration and the generation of [3H]inositol monophosphate caused by these five aggregation inducers. Platelet cyclic AMP level was unaffected by acetylshikonin. These data indicate that acetylshikonin inhibits platelet activation by suppression of phosphoinositide breakdown.

Topics: Animals; Anthraquinones; Blood Platelets; Calcium; Collagen; Cyclic AMP; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Naphthoquinones; Phosphatidylinositols; Plants, Medicinal; Platelet Aggregation; Platelet Aggregation Inhibitors; Prostaglandin D2; Prostaglandin-Endoperoxide Synthases; Rabbits; Thromboxane B2

The present study was carried out to compare the accelerative effect of shikonin (R-type), alkannin (S-type), and acetylshikonin on the proliferation of granulation tissue in rats, and to elucidate the correlation between the potency of the effect and their optical activity. Koushikon mainly contained the R-type of acetylshikonin, and Nanshikon mainly contained the S-type of acetylshikonin. Each compound produced a dose-dependent acceleration of the cotton pellet-induced granuloma formation. In comparing identical doses of shikonin, alkannin and acetylshikonin, the potency of their accelerative effects on the proliferation of granulation tissue was about the same. This result suggests that their absolute configurations (R-type or S-type) and their acetylation on the hydroxy group of the sidechain of shikonin or alkannin may not be important in producing the effect.

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents, Non-Steroidal; Cell Division; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Gossypium; Granulation Tissue; Isomerism; Male; Naphthoquinones; Rats; Rats, Wistar

Roots, root cortices and root corks from 6 species of Boraginaceae are used as zicao in commercial crude drugs. This paper reports the determination of naphthaquinone pigments (such as total pigments, beta, beta-dimethylacryshikonin, acetylshikonin and shikonin) in 12 samples of 6 plants. The quality of various drugs was evaluated by determining the contents of the above principles.

Topics: Anthraquinones; Antineoplastic Agents, Phytogenic; Chromatography, Thin Layer; Densitometry; Drugs, Chinese Herbal; Naphthoquinones; Quality Control