Naphthoquinone compound in radix arnebiae seu lithospermi as well as preparation method and application of naphthoquinone compound in preparation of medicine for treating cervical cancer

Abstract

The invention relates to the technical field of separation and purification of radix arnebiae seu lithospermi, in particular to a naphthoquinone compound in radix arnebiae seu lithospermi, a preparation method of the naphthoquinone compound and application of the naphthoquinone compound in preparation of drugs for treating cervical carcinoma, the naphthoquinone compound in radix arnebiae seu lithospermi is chemically named as (Z)-2-ethoxy-5, 8-dihydroxy-3-(4-methyl-1, 3-pentadiene-1-yl) naphthalene-1, 4-diketone, and the structural formula of the naphthoquinone compound is shown in the description. The naphthoquinone compound in the lithospermum is disclosed for the first time, in-vitro anti-tumor pharmacodynamic experiments are carried out on the naphthoquinone compound in the lithospermum, and the experiments show that the naphthoquinone compound in the lithospermum has a certain inhibition effect on human Hela cells; therefore, the naphthoquinone compound in the radix arnebiae seu lithospermi can be applied to preparation of drugs for preventing / treating cervical cancer.

Description

Technical Field

[0001] The invention relates to the technical field of separation and purification of lithospermum erythrorhizon, and specifically discloses a naphthoquinone compound in lithospermum erythrorhizon, a preparation method thereof, and application of the naphthoquinone compound in lithospermum erythrorhizon in preparing a medicine for treating cervical cancer. The chemical name of the naphthoquinone compound in lithospermum erythrorhizon is (Z)-2-ethoxy-5,8-dihydroxy-3-(4-methyl-1,3-pentadien-1-yl)naphthalene-1,4-dione. Background Art

[0002] Xinjiang Lithospermum is a plant of the Boraginaceae family ( Arnebiaeuchroma(Royle) The dried root of Lithospermum officinale (Lithospermum officinale), also known as soft lithospermum, purple dan, and ground blood, is recorded in the 2020 edition of the Chinese Pharmacopoeia as being suitable for a variety of conditions, including excessive heat and toxins in the blood, purple-black macules, subperitoneal measles, sores and eczema, and burns caused by water or fire. It has a rich chemical composition, mainly including quinones, organic acids and salts, flavonoids, polysaccharides, etc. Modern pharmacological research has shown that it has biological activities such as anti-tumor, anti-inflammatory, antiviral, anti-diabetic, weight loss and fat removal, and promotion of wound healing. A tumor refers to a neoplasm that results from the loss of normal genetic regulation of the growth of local tissue cells at the genetic level under the influence of various tumorigenic factors, leading to clonal abnormal proliferation. Currently, many anti-tumor drugs have significant side effects, and long-term use can easily lead to drug resistance. Finding highly effective and low-toxic anti-tumor drugs is the key to treating such diseases.

[0003] The naphthoquinones in Xinjiang Lithospermum have strong anti-tumor activity. Therefore, further developing and utilizing the naphthoquinones in Xinjiang Lithospermum, fully exploring their potential pharmacological value, and determining and characterizing the structural characteristics and physicochemical properties of their monomeric compounds are of great significance to promoting the development and utilization of Xinjiang Lithospermum. Summary of the Invention

[0004] The present invention provides naphthoquinone compounds in lithospermum erythrorhizon, a preparation method thereof, and application of the naphthoquinone compounds in lithospermum erythrorhizon in preparing a drug for treating cervical cancer, overcoming the deficiencies of the above-mentioned prior art. The invention discloses for the first time that the naphthoquinone compounds in lithospermum erythrorhizon have a certain inhibitory effect on HeLa cells and can be used in preparing a drug for preventing / treating cervical cancer.

[0005] One of the technical solutions of the present invention is achieved by the following measures: a naphthoquinone compound in lithospermum officinale, whose chemical name is (Z)-2-ethoxy-5,8-dihydroxy-3-(4-methyl-1,3-pentadien-1-yl)naphthalene-1,4-dione, and whose chemical structural formula is: .

[0006] The following is a further optimization and / or improvement of one of the above-mentioned technical solutions: The naphthoquinone compounds in the above-mentioned lithospermum parkii are obtained by the following method: The first step is to soak the dried aerial part of Xinjiang Lithospermum erythrorhizon in a methanol solution, then perform heating and reflux extraction to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was separated by silica gel column chromatography with gradient elution to obtain 10 fractions; In the fourth step, the second fraction of the 10 fractions obtained is subjected to silica gel chromatography gradient elution separation to obtain 8 components, the fifth component of the 8 components obtained is subjected to semi-preparative liquid chromatography separation, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

[0007] In the third step, the eluents used for silica gel column chromatography separation of the petroleum ether extract are, in order, a mixture of petroleum ether and ethyl acetate in a volume ratio of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in a volume ratio of 10:1, 5:1, and 0:1.

[0008] In the fourth step, the eluents used for silica gel column chromatography separation of the second fraction were mixtures of petroleum ether and dichloromethane in volume ratios of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1, respectively.

[0009] In the fourth step, the semi-preparative liquid chromatography separation used methanol as the eluent, the elution rate was 2 mL / min, the wavelength was 254 nm, and the chromatographic column was ODS C18, 10 mm*250 mm.

[0010] In the first step, the mass concentration of the methanol solution is 90% to 95%, and the soaking time is 40 min to 60 min.

[0011] In the first step, during the reflux extraction, the number of extractions is 2 to 4 times, and the time for each extraction is 1.0 h to 1.5 h.

[0012] The second technical solution of the present invention is achieved by the following measures: a method for preparing naphthoquinone compounds in lithospermum erythrorhizon is carried out as follows: The first step is to soak the dried aerial part of Xinjiang Lithospermum erythrorhizon in a methanol solution, then perform heating and reflux extraction to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was separated by silica gel column chromatography with gradient elution to obtain 10 fractions; In the fourth step, the second fraction of the 10 fractions obtained is subjected to silica gel chromatography gradient elution separation to obtain 8 components, the fifth component of the 8 components obtained is subjected to semi-preparative liquid chromatography separation, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

[0013] The third technical solution of the present invention is achieved through the following measures: application of naphthoquinone compounds in Lithospermum officinale in the preparation of drugs for preventing / treating cervical cancer.

[0014] The present invention discloses naphthoquinone compounds in lithospermum for the first time, and conducts in vitro antitumor pharmacodynamic experiments on the naphthoquinone compounds in lithospermum of the present invention. The experiments show that the naphthoquinone compounds in lithospermum of the present invention have a certain inhibitory effect on human Hela cells, thereby enabling the naphthoquinone compounds in lithospermum to be used in the preparation of drugs for preventing / treating cervical cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is the chemical structural formula of the naphthoquinone compound in the Lithospermum officinale of the present invention.

[0016] Figure 2 The naphthoquinone compounds in the lithospermum of the present invention 1 H-NMR spectrum.

[0017] Figure 3 The naphthoquinone compounds in the present invention are 13 C-APT spectrum.

[0018] Figure 4 HSQC spectrum of naphthoquinone compounds in Lithospermum officinale of the present invention.

[0019] Figure 5 It is the HMBC spectrum of naphthoquinone compounds in Lithospermum officinale of the present invention.

[0020] Figure 6 The naphthoquinone compounds in the lithospermum parkii prepared in Example 10 of the present invention 1 H- 1 H COSY spectrum. DETAILED DESCRIPTION

[0021] The present invention is not limited to the following embodiments, and specific implementation methods can be determined based on the technical solutions of the present invention and actual conditions. Unless otherwise specified, the various chemical reagents and chemicals mentioned in the present invention are all chemical reagents and chemicals commonly known in the prior art; unless otherwise specified, the percentages in the present invention are all percentages by mass; unless otherwise specified, the solutions in the present invention are all aqueous solutions with water as the solvent, for example, hydrochloric acid solution is an aqueous hydrochloric acid solution; normal temperature and room temperature in the present invention generally refer to temperatures between 15°C and 25°C, and are generally defined as 25°C.

[0022] The present invention will be further described below in conjunction with the embodiments: Example 1: The naphthoquinone compound in the lithospermum officinale, whose chemical name is (Z)-2-ethoxy-5,8-dihydroxy-3-(4-methyl-1,3-pentadien-1-yl)naphthalene-1,4-dione, has the chemical structural formula: .

[0023] Example 2: As an optimization of the above example, naphthoquinone compounds in Lithospermum officinale were obtained by the following method: The first step is to soak the dried aerial part of Xinjiang Lithospermum erythrorhizon in a methanol solution, then perform heating and reflux extraction to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was separated by silica gel column chromatography with gradient elution to obtain 10 fractions; In the fourth step, the second fraction of the 10 fractions obtained is subjected to silica gel chromatography gradient elution separation to obtain 8 components, the fifth component of the 8 components obtained is subjected to semi-preparative liquid chromatography separation, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

[0024] Example 3: As an optimization of the above embodiment, in the third step, the eluents used for silica gel column chromatography separation of the petroleum ether extract are, respectively, a mixture of petroleum ether and ethyl acetate in a volume ratio of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in a volume ratio of 10:1, 5:1, and 0:1.

[0025] Example 4: As an optimization of the above example, in the fourth step, the eluents used for silica gel column chromatography separation of the second fraction were mixtures of petroleum ether and dichloromethane with volume ratios of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1, respectively.

[0026] Example 5: As an optimization of the above example, in the fourth step, the eluent used in the semi-preparative liquid chromatography separation is methanol, the flow rate is 2 mL / min, the wavelength is 254 nm, and the chromatographic column is ODS C18, 10 mm*250 mm.

[0027] Example 6: As an optimization of the above example, in the first step, the mass concentration of the methanol solution is 90% to 95%, and the soaking time is 40 min to 60 min.

[0028] Example 7: As an optimization of the above example, in the first step, during reflux extraction, the number of extractions is 2 to 4 times, and the extraction time for each time is 1.0 h to 1.5 h.

[0029] Example 8: The naphthoquinone compounds in the lithospermum erythrorhizon were obtained by the following method: The first step is to add the dried aerial part of Xinjiang Lithospermum erythrorhizon to a methanol solution with a mass concentration of 90%, soak it at room temperature for 40 minutes, and then perform heating reflux extraction 3 times, each extraction for 1.0 hour to obtain an extract, and concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was sequentially separated by silica gel column chromatography using a mixture of petroleum ether and ethyl acetate in volume ratios of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in volume ratios of 10:1, 5:1, and 0:1 to obtain 10 fractions. In the fourth step, the second fraction of the obtained 10 fractions is sequentially separated by silica gel chromatography gradient elution with a mixture of petroleum ether and dichloromethane in a volume ratio of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1 to obtain 8 components, and the fifth component of the obtained 8 components is separated by semi-preparative liquid chromatography, with an eluent of 100% methanol, an elution flow rate of 2 mL / min, a wavelength of 254 nm, and a chromatographic column of ODS C18, 10 mm*250 mm, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum officinale.

[0030] Example 9: The naphthoquinone compounds in the lithospermum erythrorhizon were obtained by the following method: The first step is to add the dried aerial part of Xinjiang Lithospermum erythrorhizon to a methanol solution with a mass concentration of 95%, soak it at room temperature for 40 minutes, and then perform heating reflux extraction 3 times, each extraction for 1.5 hours to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was sequentially separated by silica gel column chromatography using a mixture of petroleum ether and ethyl acetate in volume ratios of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in volume ratios of 10:1, 5:1, and 0:1 to obtain 10 fractions. In the fourth step, the second fraction of the obtained 10 fractions is sequentially separated by silica gel chromatography gradient elution with a mixture of petroleum ether and dichloromethane with a volume ratio of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1 to obtain 8 components, and the fifth component of the obtained 8 components is separated by semi-preparative liquid chromatography, the eluent is 100% methanol, the elution flow rate is 2 mL / min, the wavelength is 254 nm, the chromatographic column is ODS C18, 10 mm*250 mm, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

[0031] Example 10: The naphthoquinone compounds in the lithospermum erythrorhizon were obtained by the following method: The first step is to add the dried aerial part of Xinjiang Lithospermum erythrorhizon to a methanol solution with a mass concentration of 95%, soak it at room temperature for 60 minutes, and then perform heating reflux extraction 3 times, each extraction for 1.5 hours to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was sequentially separated by silica gel column chromatography using a mixture of petroleum ether and ethyl acetate in volume ratios of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in volume ratios of 10:1, 5:1, and 0:1 to obtain 10 fractions. In the fourth step, the second fraction of the obtained 10 fractions is sequentially separated by silica gel chromatography gradient elution with a mixture of petroleum ether and dichloromethane with a volume ratio of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1 to obtain 8 components, and the fifth component of the obtained 8 components is separated by semi-preparative liquid chromatography, the eluent is 100% methanol, the elution flow rate is 2 mL / min, the wavelength is 254 nm, the chromatographic column is ODS C18, 10 mm*250 mm, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

[0032] Experimental Example 1: Structural characterization and analysis of naphthoquinone compounds in Lithospermum officinale of the present invention.

[0033] Experimental method: The naphthoquinone compounds in the lithospermum parkii prepared in Example 10 of the present invention were subjected to nuclear magnetic resonance spectroscopy analysis.

[0034] Experimental results: The naphthoquinone compounds in Lithospermum officinale prepared in Example 10 of the present invention 1 H-NMR spectrum Figure 2 As shown, the naphthoquinone compounds in Lithospermum officinale prepared in Example 10 of the present invention 13 C-APT spectrum Figure 3 As shown, the two-dimensional spectrum of the naphthoquinone compounds in Lithospermum officinale prepared in Example 10 of the present invention is as follows Figures 4 to 6 As shown, Figure 4 HSQC spectrum, Figure 5 is the HMBC spectrum, Figure 6 for 1 H- 1 H COSY spectrum. Figures 2 to 6 Perform graph analysis and Figures 2 to 6 The key signal peaks in the lithospermum are attributed, and the attribution of each key signal peak is shown in Table 1. According to the data in Table 1, the chemical structural formula of the naphthoquinone compounds in the lithospermum of the present invention is determined after the peaks are attributed. The chemical structural formula of the naphthoquinone compounds in the lithospermum of the present invention is as shown in Figure 1 As shown, the naphthoquinone compounds in the lithospermum erythrorhizon have good solubility in dichloromethane.

[0035] Experimental Example 2: In vitro antitumor pharmacodynamics experiment of naphthoquinone compounds in Lithospermum officinale of the present invention.

[0036] Experimental method: The naphthoquinone compounds in the lithospermum prepared in Example 10 of the present invention were subjected to an in vitro antitumor pharmacodynamics experiment. The in vitro antitumor pharmacodynamics experiment used the CCK-8 method. The naphthoquinone compounds in the lithospermum prepared in Example 10 of the present invention were used as the experimental group, and cisplatin was used as the control group. Hela (human cervical cancer cell) cells were selected as experimental subjects for the experimental group and the control group, and a blank group was set up at the same time. The specific operation was as follows: (1) Cell recovery and counting Cell recovery: Quickly remove the cryovial from the liquid nitrogen storage tank and thaw it in a 37°C water bath. Inoculate the thawed cells into a culture flask and culture in a 37°C, 5% CO2 incubator to observe whether the cells have reached the logarithmic growth phase. Use trypsin to digest the cells in the logarithmic growth phase, allowing them to detach from the flask wall and form a single-cell suspension.

[0037] Cell counting: Take an appropriate amount of cell suspension and count using a cell counting plate to ensure that the cell density is moderate.

[0038] (2) Cell suspension dilution and inoculation Cell dilution: Based on the cell count results, dilute the cells to 5 × 10 4 / mL concentration.

[0039] Cell Seeding: Add 100 μL of diluted cell suspension to each well of a 96-well plate and shake evenly to ensure even distribution of cells. Set up three replicates for each concentration of drug or compound to minimize experimental error. Also, establish a blank control group (medium alone, no cells) and a cell control group (cells and medium alone, no drug).

[0040] (3) Pre-cultivation Place the inoculated 96-well plate in a 37°C, 5% CO2 incubator for pre-culture for 24 hours to allow the cells to adhere to the wall. During the pre-culture period, observe the cell status regularly to ensure good cell growth.

[0041] (4) Drug preparation and sample addition Drug preparation: Dilute the test compound and cisplatin to the desired concentration using fresh culture medium.

[0042] Add sample: Gently add different concentrations of drug gradient to the corresponding wells to ensure that the amount of drug added is accurate and does not affect the total volume.

[0043] (5) Incubation The 96-well plate after drug addition was placed back into the incubator at 37°C and 5% CO2 for a further 48 hours and the cell status was observed regularly.

[0044] (6) Add CCK-8 reagent and measure absorbance CCK-8 solution: Mix CCK-8 reagent with fresh culture medium at a ratio of 1:10 in a dark room.

[0045] Sample addition and re-incubation: After incubation, add 10 μL of CCK-8 solution to each well. Gently shake the 96-well plate on a vortex shaker to thoroughly mix the CCK-8 solution and culture medium. Return the 96-well plate to the incubator and incubate for 2 to 4 hours.

[0046] Determine absorbance: After incubation, use a microplate reader to measure the absorbance of each well at a wavelength of 450nm. According to the formula: Inhibition rate = (A 对照 -A 实验 ) / (A 对照 -A 空白 ) to calculate the cell inhibition rate.

[0047] Each step of the experiment strictly followed the method provided by the CCK-8 kit for experiment and data processing.

[0048] Experimental results: The naphthoquinone compounds in the present invention have an IC value of Hela cells. 50 The results are shown in Table 2. It can be seen from Table 2 that the naphthoquinone compounds in the lithospermum parkii of the present invention have a certain inhibitory effect on Hela cells.

[0049] In summary, the present invention discloses naphthoquinone compounds in lithospermum for the first time, and conducts in vitro antitumor pharmacodynamic experiments on the naphthoquinone compounds in lithospermum of the present invention. The experiments show that the naphthoquinone compounds in lithospermum of the present invention have a certain inhibitory effect on human Hela cells, thereby enabling the naphthoquinone compounds in lithospermum to be used in the preparation of drugs for the prevention / treatment of cervical cancer.

[0050] The above technical features constitute the embodiments of the present invention, which have strong adaptability and implementation effect. Non-essential technical features can be added or removed according to actual needs to meet the requirements of different situations.

Claims

1. A naphthoquinone compound in Lithospermum officinale, characterized in that Its chemical name is (Z)-2-ethoxy-5,8-dihydroxy-3-(4-methyl-1,3-pentadien-1-yl)naphthalene-1,4-dione, and its chemical structure is: 。 2. The naphthoquinone compound in Lithospermum officinale according to claim 1, characterized in that Obtained as follows: The first step is to soak the dried aerial part of Xinjiang Lithospermum erythrorhizon in a methanol solution, then perform heating and reflux extraction to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was separated by silica gel column chromatography with gradient elution to obtain 10 fractions; In the fourth step, the second fraction of the 10 fractions obtained is subjected to silica gel chromatography gradient elution separation to obtain 8 components, the fifth component of the 8 components obtained is subjected to semi-preparative liquid chromatography separation, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

3. The naphthoquinone compound in Lithospermum officinale according to claim 2, characterized in that In the third step, the eluents used for silica gel column chromatography separation of the petroleum ether extract are, in order, a mixture of petroleum ether and ethyl acetate in a volume ratio of 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, and 1:1 and a mixture of dichloromethane and methanol in a volume ratio of 10:1, 5:1, and 0:

1.

4. The naphthoquinone compound in Lithospermum erythrorhizon according to claim 2 or 3, characterized in that In the fourth step, the second fraction was subjected to silica gel column chromatography using eluents of mixtures of petroleum ether and dichloromethane in volume ratios of 100:1, 50:1, 30:1, 20:1, 10:1, and 0:1, respectively.

5. The naphthoquinone compound in Lithospermum officinale according to claim 2, 3 or 4, characterized in that In the fourth step, the semi-preparative liquid chromatography separation used methanol as the eluent, the elution flow rate was 2 mL / min, the wavelength was 254 nm, and the chromatographic column was ODSC18, 10 mm*250 mm.

6. The naphthoquinone compound in Lithospermum officinale according to any one of claims 2 to 5, characterized in that In the first step, the mass concentration of the methanol solution is 90% to 95%, and the soaking time is 40 min to 60 min.

7. The naphthoquinone compound in Lithospermum officinale according to any one of claims 2 to 6, characterized in that In the first step, during reflux extraction, the extraction times are 2 to 4 times, and the extraction time for each time is 1.0 h to 1.5 h.

8. A method for preparing naphthoquinone compounds from Lithospermum officinale according to any one of claims 1, 3 to 7, characterized in that Proceed as follows: The first step is to soak the dried aerial part of Xinjiang Lithospermum erythrorhizon in a methanol solution, then perform heating and reflux extraction to obtain an extract, and then concentrate the extract under reduced pressure to obtain a Lithospermum erythrorhizon concentrate; In the second step, the obtained Lithospermum erythrorhizon concentrated solution is extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extracts are concentrated to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; In the third step, the obtained petroleum ether extract was separated by silica gel column chromatography with gradient elution to obtain 10 fractions; In the fourth step, the second fraction of the 10 fractions obtained is subjected to silica gel chromatography gradient elution separation to obtain 8 components, the fifth component of the 8 components obtained is subjected to semi-preparative liquid chromatography separation, and the eluate is collected to obtain naphthoquinone compounds in Lithospermum erythrorhizon.

9. Use of the naphthoquinone compound in Lithospermum officinale according to any one of claims 1 to 7 in the preparation of a drug for preventing and / or treating cervical cancer.

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