• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.293-299
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• 2003

The effects of low dosage ${\gamma}$-radiation on the cell growth and the formation of shikonin derivatives were investigated in callus cultures of Lithospermum erythrorhizon under different medium and light conditions. Gamma radiation significantly affected the cell growed and formation of shikonin derivatives, depending on the culture conditions. In the cell cultures grown on M-9 medium, 2Gy and 16Gy of ${\gamma}$-radiation increased the calli growth and the formation of shikonin derivatives, respectively under 16hr day light condition. When calli were cultured for 60 days in the dark after irradiation of ${\gamma}$-radiation, cell growth was increased at low dosage of 1Gy and 2Gy in LS medium containing BA 2mg/L and IAA 0.2mg/L. Interestingly, calli grown in M-9 medium by 2Gy irradiation for 60 days significantly stimulated the formation of shikonin derivatives(13.21mg/g cell fresh wt), which was approximately 6 times higher than untreated cells.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.565-569
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• 1998

An HPLC method was developed to resolve the enantiomers of shikonin derivatives of the Lithospermi Radix. The optimum mobile phase on a Chiracel AD column was 5% isopropanol in n-hexane with folw rate of 1 ml/min. Establishment of this method made possible to determine the ratios of shikonin/acetylshikonin or alkanin/acetylalkanin in the same root. The correlation of the ratios of these substance pairs appeared characteristic for the country where they were originated from. All of the Korean species showed significantly higher ratios of shikonin/acetylshikonin and alkanin/acetylalkanin than the Chinese ones. this method would be useful to determine the orgin of Lithospermi Radix.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.343-348
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• 1989

Production of shikonin derivatives by Lithospermum erythrorhizon cells by using polyurethane foam was invesliigated. Shikonin derivatives were effectively adsorbed mostly by phase distribution to polyurethane matrices and their production increased significantly compared to the suspension culture. The enhanced production of shikonin was probably due to more facilitated cell to cell con-tact and lowered intracellular shikonin concentration, both of which are known to be favorable for plant secondary metabolite production. In order to improve the process productivity, tell culture was conducted under various culture conditions: Of them, Schenk and Hildebrandt medium containing indole-3-acetic acid (1.75mg/ι) and kinetin (0.1mg/ι) was considered most appropriate for shikonin production. Production of shikonin increased about 4.5 times in the Schenk and Hildebrandt medium containing indole-3-acetic acid (1.15mg/ι) and kinetin (0.1mg/ι) when compared to the same medium containing p-chlorophenoxyacetic acid (2.0mg/ι) and kinetin (0.1mg/ι). When poly-urethane was used as the support material, a single-stage system was more preferred to the conventional two-stage culture system in terms of shikonin productivity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.2
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• pp.177-181
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• 2006

In this study, phytochemicals from the wold and cultivated Lithospermum erythrorhizon (gromwell), which has been used for medicinal purpose or natural coloring material from the old days, were extracted by methanol and fractionated with hexane. The shikonin compounds in the fraction was isolated and their chemical structures were identified by $^1H$ and $^{13}C-NMR$. It was found that compound I was the shikonin substance with molecular weight of 288.3 and chemical formula of $C_{16}H_{16}O_5$, and compound II being deoxyshikonin substance with molecular weight of 272.3 and chemical formula of $C_{16}H_{16}O_4$. The Quantities of these compounds in the wild and cultivated gromwells was determined.

• Archives of Pharmacal Research
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• v.19 no.5
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• pp.416-422
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• 1996

Twenty one phenylacetylshikonin analogues were synthesized from various subsitituted phenyl acetic acids and their cytotoxicity values against A549, K562 and L1210 cell lines and antitumor action in mice bearing S-180 cells were measured. All of phenylacetylshikonin analogues expressed a potent cytotoxicity $(ED_{50}, 0.1-1.80{\mu}g/ml)$ against L1210 and K562 cells. L1210 cells were the most sensitive to shikonin analogues among these cells. Except 4-methosyphenylacetylshikonin $(0.098 {\mu}g/ml)$, and a-acetoxyphenylacetylshikonin $(0.10 {\mu}g/ml)$, all other shikonin derivatives sshowed higher $ED_{50}$ values than phenylacetylshikonin $(0.13{\mu}g/ml)$, in L1210. In K562 cell, a-substitution of phenylacetylshikonin $(0.1{\mu}g/ml)$, while other subsitutions increased it slightly; 4-methoxyphenylacetylshikonin $(0.033{\mu}g/ml)$ showed a exceptionally good cytotoxicity against K562 cell. 4-Halogenation tended to decrease the cytotoxic effect on L1210 cells, while it enhanced the effect on K562; 4-bromophenylacetyl $$[ED_{50};(L1210)=1.76{\mu}g/ml, ;ED_{50};(K 562)=0.32 {\mu}g/ml]$$ and 4-chlorophenylacetyl shikonin $$[ED_{50};(L1210)=1.64 {\mu}g/ml, ;ED_{50};(K562)=0.32 {\mu}g/ml]$$. In contrast, A549 cells were much less sensitive to these shikonin analogues which showed $ED_{50}$ values of$1.5-1.35 {\mu}g/ml)$.Most of phenylacetylshikonin derivatives showed good antitumor activity in mice bearing S-180 cells. a-A-cetoxyphenylacetylshikonin and 4-dimethylaminophenylacetylshikonin showed highest T/C value (192-195%), implying that introduction of a-acetyl or of 4-dimethylamino group enhanced the antitumor activity as shown for 4-dimethylaminophenylacetylshikonin (T/C, 192%). It might be due to improvement of water solubility by dimethylamino group in the molecule.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.13 no.1
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• pp.7-14
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• 1987

Production of shikonin derivatives through cell suspension culture of Lithospermum erythrorhizon was investigated. Optimal concentrations of IAA and kinetin on the growth of cell suspension were 0.2 and 0.1 ppm respectively. Pigment content was markedly increased when aluminum oxide was added to the production medium and its optimal concentration was 1.5g/70ml medium. The most effective concentration of IAA was 0.5 ppm and the production of pigment did not depend on the kinetin concentration.

• Korean Journal of Pharmacognosy
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• v.41 no.2
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• pp.73-88
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• 2010

Traditionally, the root of Lithospermum erythrorhizon Sieb. et Zucc(L.E) has been used as efficacious therapy for inflammation, burns, frostbite and skin ailments (e.g eczema and psoriasis). It contains isohexenylnaphthoquinone derivatives (shikonin and its esters) and furylhydroquinones (shikonofurans) in lipophilic fractions and caffeic acid oligomers (rosmarinic acid, lithospermic acid B) in polar fractions. Recently, new preparative isolation and analysis procedures of shikonin along with its oligomers from the extract of L. erythrorhizon by the combination of high-speed counter-current chromatography with high-performance liquid chromatography-diode array detection have also been introduced. Although there have been many reports on the wound healing, antiinflammatory, and anticancer effects, the research on the effects of anti-atopic dermatitis of the root of L. erythrorhizon were relatively scarce. However, in recent years, new information gathered from research efforts, on the anti-atopic dermatitis properties of the extract or constituents of L. erythrorhizon has been accumulated. In this paper, the findings and advance on the in vitro and in vivo activities of L. erythrorhizon and its constituents especially focused on antiinflammatory and anti-atopic dermatitis effects are summarized. The phytochemical constituents of L. erythrorhizon or its tissue cultures are also presented. Although there are few to verify or refute its activity in human, one result of clinical study of the extract of L. erythrorhizon on the atopic dermatitis patients was introduced to assess the possibility of its clinical use. The reported mechanisms of action and in vivo pharmacological studies in different animal models for the various types of extracts or constituents of L. erythrorhizon are supportive of its therapeutic potential or dietary supplement, however, more evidence from clinically relevant models, as well as systemic studies on the active constituents or the various types of standardized extracts at the cellular and molecular level, are required.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.208.1-208.1
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• 2003

We synthesized naphthazarin derivatives from shikonin, a major compound from Lithospermum erythrorhion Sieb et ZUCC. Of derivatives, DMNQ S64, 2-or 6-(l-hydroxyiminoalkyl) effectively showed antitumor activity on A549, human lung cancer cells (IC$\sub$50/= 30 ${\mu}$M). It significantly inhibited prostaglandin E$_2$(IC$\sub$50/= 10 ${\mu}$M). We also confirmed it selectively downregulated the expression of cyclooxygenase 2(COX-2), while it didn't affect COX-1. The induction of apoptosis by DMNQ S64 is underway.

• Journal of Life Science
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• v.27 no.3
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• pp.325-333
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• 2017

High level of plasma cholesterol is strongly associated with the development of atherosclerosis and coronary heart disease. Clinical trials designed to reduce plasma cholesterol level by diet or pharmacological intervention have resulted in marked reduction of disease incidence. The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase which reduces cholesterol biosynthesis in the liver is the key enzyme of the mevalonate pathway that produces cholesterol. In this study, 71 naturally occurring phenolic compounds were tested for inhibitory activities against HMG-CoA reductase. Eleven compounds out of 71 showed inhibitory activities: three hydrolyzable tannin (geraniin, acetonyl geraniin and pentagalloyl ${\beta}-D-glucose$), four benzoic acid derivatives (benzoic acid, trans-cinnamic acid, 2,4-dihydroxybenzoic acid and 2,5-dihydroxybenzoic acid), and four naphthoquinone derivatives (1,2-naphthoquinone, 1,4-naphthoquinone, plumbagin and shikonin). At the concentration of $10{\mu}g/ml$, 1,4-naphthoquinone inhibited HMG-CoA reductase by 99.4%, and then plumbagin 91.4%, pentagalloyl ${\beta}-D-glucose$ 46.6%, 2,4-dihydroxybenzoic acid 40.9%, shikonin 37.7%, 1,2-naphthoquinone 36.6%, trans-cinnamic acid 32.0%, acetonyl geraniin 30.2%, benzoic acid 28.5%, geraniin 28.3% and 2,5-dihydroxybenzoic acid 22.3%, respectively. $IC_{50}$ values of 1,4-naphthoquinone and plumbagin was $2.1{\mu}g/ml$ and $5.8{\mu}g/ml$, respectively.

• YAKHAK HOEJI
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• v.42 no.2
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• pp.159-164
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• 1998

The secondary hydroxy group at side chain of shikonin structure was selectively acylated with various haloacetic acids in presence of dicyclohexylcarbodiimide and 4-dimethylamin opyridine to produce haloacetylshikonin derivatives. The cytotoxicity of monohaloacetylshikonin derivatives against L1210 cells increased in the following order; monochloroacetylshikonin ($ED_{50}$, 0.142${\mu}$g/ml) > monobromoacetylshikonin ($ED_{50}$. 0.158${\mu}$g/ml) > monoiodoacetylshikonin ($ED_{50}$, 0.173${\mu}$g/ml). Introduction of larger halogen atoms decreased the cytotoxic activity, presumably due to steric hinderance. The cytotoxicity of chloroacetylshikonin derivatives was dependent on the number of chlorine atom, thus increasing in the following order; trichloroacetylshikonin (0.032${\mu}$g/ml) > dichloroacetylshikonin (0.059${\mu}$g/ml) > monochloroacetylshikonin ($ED_{50}$, 0.142${\mu}$g/ml). Thus, the electron withdrawing effect seems to be important for the cytotoxicity of chloroacetylshikonin derivatives. Consistent with the above, dichloroacetylshikonin (T/C. 182%) and trifluoroacetylshikonin (195%) showed higher T/C values than monochloroacetyl-(T/C, 122%), monobromoacetyl-(T/C, 154%) and monoiodoacetylshikonin (T/C, 117%) derivatives. Haloacetylshikonin derivatives showing lower cytoxic activities against L1210 cells exhibited lower T/C values. It seems that there is a relationship between the cytoxicity of haloacetylshikonin and their antitumor activity.