Application of phenolic compound and / or pharmaceutically acceptable salt thereof in preparation of medicine for treating breast cancer, and pharmaceutical composition

Abstract

The invention provides application of a phenolic compound and / or pharmaceutically acceptable salt thereof in preparation of a medicine for treating breast cancer and a pharmaceutical composition, and relates to the technical field of biological medicines. The invention discloses application of phenolic compounds and / or pharmaceutically acceptable salts thereof in preparation of medicines for treating breast cancer. The phenolic compounds comprise a compound 1 and / or a compound 2. The compound 1 (arnebidin) and the compound 2 ((+)-(R)-de-O-metalasiodiplodin) adopted in the invention have good inhibition effects on human triple-negative breast cancer MDA-MB-231 cell strains, the IC50 value of the compound 1 is 1.16 [mu] M, and the IC50 value of the compound 2 is 0.68 [mu] M, so that a new choice is provided for developing drugs for treating triple-negative breast cancer, and the compound 1 and the compound 2 have good application prospects. The compound has a good application prospect in preparation of medicines for treating breast cancer, especially medicines for treating triple-negative breast cancer.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of phenolic compounds and / or pharmaceutically acceptable salts thereof in preparing drugs for treating breast cancer and a pharmaceutical composition. Background Art

[0002] Malignant tumors are common and frequently occurring diseases that seriously threaten human health and life. Among them, breast cancer, as the malignant tumor with the highest incidence and mortality rate among women worldwide, has always received widespread attention. Triple-negative breast cancer is a special type of breast carcinoid, with an incidence of about 15-20% of all breast cancers. Compared with other breast cancer subtypes, triple-negative breast cancer lacks the immunohistochemical expression of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor-2. It is more invasive, develops rapidly, has a high recurrence rate, and has a poor prognosis. It is the most malignant type of breast cancer. Triple-negative breast cancer lacks suitable therapeutic targets, and the commonly used endocrine therapy and targeted therapy are relatively ineffective. The available treatments are relatively limited. At present, there is an urgent need to research and develop drugs that can effectively inhibit the activity of triple-negative breast cancer. Summary of the invention

[0003] In view of this, the object of the present invention is to provide the use of phenolic compounds and / or pharmaceutically acceptable salts thereof in the preparation of drugs for treating breast cancer and pharmaceutical compositions. Compounds 1 and 2 used in the present invention have high inhibitory activity against triple-negative breast cancer.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] The present invention provides the use of phenolic compounds and / or pharmaceutically acceptable salts thereof in preparing a drug for treating breast cancer, wherein the phenolic compounds include compound 1 and / or compound 2:

[0006]

[0007] Preferably, the breast cancer comprises triple-negative breast cancer.

[0008] Preferably, the pharmaceutically acceptable salt comprises a sodium salt or a potassium salt.

[0009] Preferably, the preparation method of the phenolic compound comprises the following steps:

[0010] The root of Lithospermum erythrorhizon is extracted by using a low-grade alcohol aqueous solution to obtain an alcohol extract;

[0011] The alcohol extract is subjected to a first silica gel column chromatography to collect the third and fourth components, which are respectively recorded as Fr.3 and Fr.4; the elution mode of the first silica gel column chromatography is gradient elution, and the eluents used in the gradient elution include petroleum ether-ethyl acetate and ethyl acetate-methanol in sequence; the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate is 98:2, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 25:75 and 0:100; the volume ratio of ethyl acetate to methanol in the ethyl acetate-methanol is 90:10, 80:20, 50:50 and 0:100 in sequence;

[0012] The preparation method of compound 1 comprises the following steps:

[0013] The Fr.3 is subjected to a second silica gel column chromatography, and the first component is collected and recorded as Fr.3.1; the eluent used in the second silica gel column chromatography is petroleum ether-ethyl acetate; the elution method of the second silica gel column chromatography is gradient elution, and the volume ratios of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate during the second silica gel column chromatography are 80:1, 60:1, 30:1, 15:1 and 0:100 respectively;

[0014] The Fr.3.1 is subjected to a first gel column chromatography to collect the second component, which is recorded as Fr.3.1.2; the Fr.3.1.2 is subjected to a second gel column chromatography to collect the second component, which is recorded as Fr.3.1.2.2; the eluents used in the first gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the first gel column chromatography process is 20-80%; the eluent used in the second gel column chromatography is methanol;

[0015] The Fr.3.1.2.2 is separated by a first reverse phase chromatographic column to obtain compound 1; the eluent used for separation by the first reverse phase chromatographic column is methanol-water;

[0016] The preparation method of the compound 2 comprises the following steps:

[0017] The Fr.4 is subjected to a third silica gel column chromatography, and the second component is collected and recorded as Fr.4.2; the eluents used in the third silica gel column chromatography are dichloromethane and methanol; the elution method of the third silica gel column chromatography is gradient elution, and the volume ratios of dichloromethane and methanol in the third silica gel column chromatography are 100:1, 80:1, 50:1, 25:1, 10:1 and 0:100 respectively;

[0018] The Fr.4.2 is subjected to a third gel column chromatography to collect the fourth component, which is recorded as Fr.4.2.4; the Fr.4.2.4 is subjected to a fourth gel column chromatography to collect the third component, which is recorded as Fr.4.2.4.3; the eluents used in the third gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the third gel column chromatography process is 20-80%; the eluent used in the fourth gel column chromatography is methanol;

[0019] The Fr.4.2.4.3 is subjected to a second reverse phase chromatography column separation to obtain compound 2; the eluent used for the second reverse phase chromatography column separation is methanol-water.

[0020] Preferably, the volume fraction of the lower alcohol in the lower alcohol aqueous solution is 50-100%; the lower alcohol includes methanol and / or ethanol;

[0021] The volume ratio of the dry weight of the lithospermum root to the lower alcohol aqueous solution for single extraction is 4-6 kg: 6-7 L;

[0022] The extraction temperature is 15-40° C., the number of extractions is 2-4 times, and the time for a single extraction is 24-120 hours.

[0023] Preferably, the gels used in the first gel column chromatography, the second gel column chromatography, the third gel column chromatography and the fourth gel column chromatography include hydroxypropyl dextran gel.

[0024] Preferably, the volume fraction of methanol in the methanol-water used for separation in the first reverse phase chromatography column is 85-95%;

[0025] The volume fraction of methanol in methanol-water used for separation in the second reverse phase chromatography column is 86-90%.

[0026] The present invention also provides a pharmaceutical composition, comprising an active component and a pharmaceutically acceptable excipient; the active component is a phenolic compound and / or a pharmaceutically acceptable salt thereof, and the phenolic compound is the phenolic compound in the application described in the above technical solution.

[0027] Preferably, the content of the active ingredient in the pharmaceutical composition is 0.1 to 99 wt%.

[0028] Preferably, the dosage form of the pharmaceutical composition includes oral preparation or injection.

[0029] Compound 1 (arnebidin) and compound 2 ((+)-(R)-de-O-methyllasiodiplodin) used in the present invention have good inhibitory effects on human triple-negative breast cancer MDA-MB-231 cell line. The IC 50 The value is 1.16 μM, and the IC of compound 250 The value is 0.68 μM, which provides a new option for the development of therapeutic drugs for triple-negative breast cancer and has a good application prospect in the preparation of drugs for treating breast cancer, especially triple-negative breast cancer.

[0030] The present invention first extracts the lithospermum root by cold soaking with a low-grade alcohol aqueous solution, and then separates it by silica gel column chromatography, gel chromatography and reverse phase chromatography column separation to obtain compound 1 and compound 2. In the present invention, silica gel column chromatography is simple to operate, has obvious separation effect and large sample carrying capacity; gel chromatography has good separation degree, fast separation speed and convenient operation; reverse phase chromatography column separation method has flexible sample quantity and can process samples from micrograms to milligrams, has high separation efficiency, and prepares compounds 1 and 2 with high purity. Compounds 1 and 2 used in the present invention have good inhibitory activity effect on human triple negative breast cancer MDA-MB-231 cells. DETAILED DESCRIPTION

[0031] The present invention provides the use of phenolic compounds and / or pharmaceutically acceptable salts thereof in preparing a drug for treating breast cancer, wherein the phenolic compounds include compound 1 and / or compound 2:

[0032]

[0033] In the present invention, the English name of the compound 1 is arnebidin; the English name of the compound 2 is (+)-(R)-de-O-methyllasiodiplodin.

[0034] In the present invention, the breast cancer includes triple-negative breast cancer.

[0035] In the present invention, the pharmaceutically acceptable salt preferably includes a sodium salt or a potassium salt. In the present invention, the pharmaceutically acceptable salt is preferably prepared by a phenolic compound and a metal hydroxide through a salt-forming reaction; the salt-forming site of the phenolic compound is a phenolic hydroxyl group. In the present invention, the metal hydroxide is preferably sodium hydroxide or potassium hydroxide. The present invention has no special restrictions on the conditions of the salt-forming reaction, and the conventional salt-forming reaction conditions in the art can be used.

[0036] In the present invention, the preparation method of the phenolic compound comprises the following steps:

[0037] The root of Lithospermum erythrorhizon is extracted by using a low-grade alcohol aqueous solution to obtain an alcohol extract;

[0038] The alcohol extract is subjected to a first silica gel column chromatography to collect the third and fourth components, which are respectively recorded as Fr.3 and Fr.4; the elution mode of the first silica gel column chromatography is gradient elution, and the eluents used in the gradient elution include petroleum ether-ethyl acetate and ethyl acetate-methanol in sequence; the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate is 98:2, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 25:75 and 0:100; the volume ratio of ethyl acetate to methanol in the ethyl acetate-methanol is 90:10, 80:20, 50:50 and 0:100 in sequence;

[0039] The preparation method of compound 1 comprises the following steps:

[0040] The Fr.3 is subjected to a second silica gel column chromatography, and the first component is collected and recorded as Fr.3.1; the eluent used in the second silica gel column chromatography is petroleum ether-ethyl acetate; the elution method of the second silica gel column chromatography is gradient elution, and the volume ratios of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate during the second silica gel column chromatography are 80:1, 60:1, 30:1, 15:1 and 0:100 respectively;

[0041] The Fr.3.1 is subjected to a first gel column chromatography to collect the second component, which is recorded as Fr.3.1.2; the Fr.3.1.2 is subjected to a second gel column chromatography to collect the second component, which is recorded as Fr.3.1.2.2; the eluents used in the first gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the first gel column chromatography process is 20-80%; the eluent used in the second gel column chromatography is methanol;

[0042] The Fr.3.1.2.2 is separated by a first reverse phase chromatographic column to obtain compound 1; the eluent used for separation by the first reverse phase chromatographic column is methanol-water;

[0043] The preparation method of the compound 2 comprises the following steps:

[0044] The Fr.4 is subjected to a third silica gel column chromatography, and the second component is collected and recorded as Fr.4.2; the eluents used in the third silica gel column chromatography are dichloromethane and methanol; the elution method of the third silica gel column chromatography is gradient elution, and the volume ratios of dichloromethane and methanol in the third silica gel column chromatography are 100:1, 80:1, 50:1, 25:1, 10:1 and 0:100 respectively;

[0045] The Fr.4.2 is subjected to a third gel column chromatography to collect the fourth component, which is recorded as Fr.4.2.4; the Fr.4.2.4 is subjected to a fourth gel column chromatography to collect the third component, which is recorded as Fr.4.2.4.3; the eluents used in the third gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the third gel column chromatography process is 20-80%; the eluent used in the fourth gel column chromatography is methanol;

[0046] The Fr.4.2.4.3 is subjected to a second reverse phase chromatography column separation to obtain compound 2; the eluent used for the second reverse phase chromatography column separation is methanol-water.

[0047] Unless otherwise specified, the materials and equipment used in the present invention are all commercially available products in the art.

[0048] The invention utilizes a low-grade alcohol aqueous solution to extract the lithospermum root to obtain an alcohol-extracted extract.

[0049] In the present invention, the lithospermum root is preferably dried before being cut before use. In the present invention, the drying temperature is preferably 20 to 35° C., more preferably 25° C.; the drying time is preferably 8 to 20 days, more preferably 10 to 15 days. In the present invention, the length of the cut lithospermum root is preferably 0.5 to 2 cm, more preferably 1 to 1.5 cm.

[0050] In the present invention, the volume fraction of the lower alcohol in the lower alcohol aqueous solution is preferably 50-100%. In a specific embodiment, the volume fraction of the lower alcohol in the lower alcohol aqueous solution can be 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%. The lower alcohol preferably includes methanol and / or ethanol, more preferably methanol.

[0051] In the present invention, the volume ratio of the dry weight of the lithospermum root to the lower alcohol aqueous solution is preferably 4-6kg:6-7L. In a specific embodiment, the volume ratio of the dry weight of the lithospermum root to the lower alcohol aqueous solution for single extraction can be 4kg:6L, 4.5kg:6L, 5kg:6L, 5.5kg:6L, 6kg:6L, 4kg:7L, 4.5kg:7L, 5kg:7L, 5.5kg:7L, 6kg:7L, 4kg:8L, 4.5kg:8L, 5kg:8L, 5.5kg:8L or 6kg:8L.

[0052] In the present invention, the extraction temperature is preferably 15-40°C, more preferably 20-30°C; the number of extractions is preferably 2-4 times, more preferably 3 times; the time for a single extraction is preferably 24-120h. In a specific embodiment, the single extraction time can be 24h, 30h, 40h, 50h, 60h, 70h, 80h, 90h, 100h, 110h or 120h.

[0053] After the extraction is completed, the present invention preferably further comprises combining the obtained extracts and concentrating them to obtain an alcohol extract. The present invention has no special limitation on the concentration, and a concentration method well known to those skilled in the art can be used until the volume no longer changes, such as reduced pressure concentration.

[0054] After obtaining the alcohol extract, the present invention performs the first silica gel column chromatography on the alcohol extract, collects the third and fourth components respectively, and records them as Fr.3 and Fr.4 respectively; the elution mode of the first silica gel column chromatography is preferably gradient elution, and the eluents used in the gradient elution preferably include petroleum ether-ethyl acetate and ethyl acetate-methanol in sequence; the volume ratio of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate is preferably 98:2, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 25:75 and 0:100; the volume ratio of ethyl acetate and methanol in the ethyl acetate-methanol is preferably 90:10, 80:20, 50:50 and 0:100 in sequence; in the present invention, the particle size of the silica gel used in the first silica gel column chromatography is preferably 200-300 meshes. In the present invention, the first silica gel column chromatography obtains 12 components, which are recorded as Fr.1-Fr.12.

[0055] The preparation method of the compound 1 is described below.

[0056] After obtaining Fr.3, the present invention performs a second silica gel column chromatography on Fr.3 to collect the first component, which is recorded as Fr.3.1. In the present invention, the eluent used in the second silica gel column chromatography is preferably petroleum ether-ethyl acetate; the elution method of the second silica gel column chromatography is preferably gradient elution, and the volume ratio of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate during the second silica gel column chromatography is preferably 30:1, 10:1, 5:1, 2:1 and 0:100 respectively; the particle size of the silica gel used in the second silica gel column chromatography is preferably 200-300 meshes. In the present invention, the second silica gel column chromatography obtains 6 components, which are recorded as Fr.3.1-Fr.3.6.

[0057] After obtaining Fr.3.1, the present invention performs a first gel column chromatography on Fr.3.1 to collect the second component, which is recorded as Fr.3.1.2. In the present invention, the eluents used in the first gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the first gel column chromatography process is 20-80%. In a specific embodiment, the volume fraction of dichloromethane in the first gel column chromatography process can be 20%, 30%, 40%, 50%, 60%, 70% or 80%; the gel used in the first gel column chromatography preferably includes hydroxypropyl dextran gel (Sephadex LH-20); the elution mode of the first gel column chromatography is preferably isocratic elution.

[0058] After obtaining Fr.3.1.2, the present invention performs a second gel column chromatography on Fr.3.1.2 to collect the second component, which is recorded as Fr.3.1.2.2. In the present invention, the eluent used in the second gel column chromatography preferably includes methanol; and the gel used in the second gel column chromatography preferably includes hydroxypropyl dextran gel.

[0059] After obtaining Fr.3.1.2.2, the present invention performs a first reverse phase chromatography column separation on Fr.3.1.2.2 to obtain compound 1. In the present invention, the eluent for separation by the first reverse phase chromatography column is preferably methanol-water, and the volume fraction of methanol in the methanol-water for separation by the first reverse phase chromatography column is preferably 85-95%. In a specific embodiment, the volume fraction of methanol in the methanol-water for separation by the first reverse phase chromatography column can be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% or 95%.

[0060] The preparation method of compound 2 is described in detail below.

[0061] After obtaining Fr.4, the present invention performs a third silica gel column chromatography on Fr.4 to collect the second component, which is recorded as Fr.4.2. In the present invention, the eluent used in the third silica gel column chromatography is preferably dichloromethane and methanol; the elution method of the third silica gel column chromatography is preferably gradient elution, and the volume ratio of dichloromethane and methanol in the third silica gel column chromatography is preferably 100:1, 80:1, 50:1, 25:1, 10:1 and 0:100 respectively; the particle size of the silica gel used in the third silica gel column chromatography is preferably 200-300 meshes. In the present invention, the third silica gel column chromatography obtains 5 components, which are recorded as Fr.4.1-Fr.4.5.

[0062] After obtaining Fr.4.2, the present invention performs a third gel column chromatography on Fr.4.2 to collect the fourth component, which is recorded as Fr.4.2.4. In the present invention, the eluent used in the third gel column chromatography is preferably dichloromethane and methanol, and the volume fraction of dichloromethane in the third gel column chromatography process is preferably 20-80%. In a specific embodiment, the volume fraction of dichloromethane in the third gel column chromatography process can be 20%, 30%, 40%, 50%, 60%, 70% or 80%; the gel used in the third gel column chromatography preferably includes hydroxypropyl dextran gel; the elution method of the third gel column chromatography is preferably isocratic elution.

[0063] After obtaining Fr.4.2.4, the present invention performs a fourth gel column chromatography on Fr.4.2.4 to collect the third component, which is recorded as Fr.4.2.4.3. In the present invention, the eluent used in the fourth gel column chromatography is preferably methanol; and the gel used in the fourth gel column chromatography preferably includes hydroxypropyl dextran gel.

[0064] After obtaining Fr.4.2.4.3, the present invention performs a second reverse phase chromatography column separation on Fr.4.2.4.3 to obtain compound 2. In the present invention, the eluent for separation by the second reverse phase chromatography column is preferably methanol-water; the volume fraction of methanol in the methanol-water for separation by the second reverse phase chromatography column is preferably 86-90%, and in a specific embodiment, the volume fraction of methanol in the methanol-water for separation by the second reverse phase chromatography column can be 86%, 86.5%, 87%, 87.5%, 88%, 88.5%, 89%, 89.5% or 90%.

[0065] The present invention also provides a pharmaceutical composition, comprising an active component and a pharmaceutically acceptable excipient; the active component is a phenolic compound and / or a pharmaceutically acceptable salt thereof, and the phenolic compound is the phenolic compound in the application described in the above technical solution.

[0066] In the present invention, the content of the active ingredient in the pharmaceutical composition is preferably 0.1 to 99wt%. In a specific embodiment, the content of the active ingredient in the pharmaceutical composition can be 0.1wt%, 0.5wt%, 1wt%, 5wt%, 10wt%, 15wt%, 20wt%, 25wt%, 30wt%, 35wt%, 40wt%, 45wt%, 50wt%, 55wt%, 60wt%, 65wt%, 70wt%, 75wt%, 80wt%, 85wt%, 90wt%, 95wt% or 99wt%.

[0067] In the present invention, the dosage form of the pharmaceutical composition preferably includes an oral agent or an injection. In the present invention, the oral agent preferably includes a powder, tablet, sugar-coated agent, capsule, pill or solution syrup; the injection preferably includes a powder injection or a solution injection.

[0068] The pharmaceutically acceptable excipients of the present invention preferably include pharmaceutical carriers and / or excipients. In the present invention, the pharmaceutically acceptable excipients preferably include one or more of lactose, starch, magnesium stearate and sodium chloride solution. In the present invention, the pharmaceutically acceptable excipients are non-toxic to humans and animals and are inert excipients.

[0069] In the present invention, the preparation method of the tablet preferably comprises the following steps: mixing the active ingredient, lactose and starch to obtain a mixture; wetting the mixture with water and then sieving it to obtain the undersize; and tableting the undersize with magnesium stearate for the second time to obtain a tablet. In the present invention, the mass ratio of the active ingredient to lactose is preferably 1:17-20. In a specific embodiment, the mass ratio of the active ingredient to lactose can be 1:17, 1:17.5, 1:18, 1:18.5, 1:19, 1:19.5 or 1:20; the mass ratio of the active ingredient to starch is preferably 1:5.3-5.8. In a specific embodiment, the mass ratio of the active ingredient to starch can be 1:5.3, 1:5.35, 1:5.4, 1:5.45, 1:5.5, 1:5.55, 1:5.6, 1:5.65, 1:5.7, 1:5.75 or 1:5.8. The present invention has no special requirements for the first mixing, as long as it can be mixed evenly. In the present invention, the mass ratio of the active component and magnesium stearate is preferably 1: 0.4-0.7. In a specific embodiment, the mass ratio of the active component and magnesium stearate can be 1: 0.4, 1: 0.45, 1: 0.5, 1: 0.55, 1: 0.6, 1: 0.65 or 1: 0.7. In the present invention, the water is preferably distilled water or sterile water, more preferably distilled water. In the present invention, the aperture of the sieve for sieving is preferably 75-85 mesh, more preferably 80 mesh. The present invention has no special requirements for the second mixing, as long as it can be mixed evenly. The present invention has no special requirements for the tableting, and the conventional method in the art can be used.

[0070] In the present invention, the specification of the tablet is preferably 240-260 mg / tablet, more preferably 250 mg / tablet, and the content of the active ingredient in the tablet is preferably 9-11 mg, more preferably 10 mg.

[0071] In the present invention, the preparation method of the capsule preferably comprises the following steps: the active component and starch are mixed and then sieved, and the sieve material is filled into a magnesium stearate capsule shell to obtain a capsule.

[0072] In the present invention, the mass ratio of the active component to the starch is preferably 0.8 to 1.2:1. In a specific embodiment, the mass ratio of the active component to the starch can be 0.8:1, 0.85:1, 0.9:1, 0.95:1, 1:1, 1.05:1, 1.1:1, 1.15:1 or 1.2:1. The present invention has no special requirements for the mixing, as long as it can be mixed. In the present invention, the aperture of the sieve for screening is preferably 75 to 85 mesh, more preferably 80 mesh.

[0073] In the present invention, the preparation method of the injection ampoule preferably comprises the following steps: dissolving the active component and sodium chloride in water for injection, filtering, and filling the filtrate obtained by the filtration into an ampoule bottle under sterile conditions and sealing to obtain an injection ampoule. In the present invention, the mass ratio of the active component to sodium chloride is preferably 1:8-11. In a specific embodiment, the mass ratio of the active component to sodium chloride may be 1:8, 1:8.5, 1:9, 1:9.5, 1:10, 1:10.5 or 1:11. In the present invention, the mass ratio of the active component to the volume ratio of water for injection is preferably 2mg:90-110mL. In a specific embodiment, the mass ratio of the active component to the volume ratio of water for injection may be 2mg:90mL, 2mg:95mL, 2mg:100mL, 2mg:105mL or 2mg:110mL.

[0074] In the present invention, the pharmaceutical composition is preferably used in the form of a dosage per unit body weight, and the administration method is preferably oral administration and / or injection, and the injection preferably includes intravenous injection or intramuscular injection.

[0075] In order to further illustrate the present invention, the use of the phenolic compounds and / or pharmaceutically acceptable salts thereof provided by the present invention in the preparation of drugs for treating breast cancer and the pharmaceutical compositions are described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0076] Example 1

[0077] S1: 50 kg of lithospermum root was dried at 25° C. for 10 days and then cut into small segments with a length of 1 cm; the cut lithospermum root was immersed in a mixed solvent of 95% methanol and water at 30° C. for extraction, and the extraction was repeated 3 times, the amount of the mixed solvent of methanol and water used for a single immersion extraction was 60 L, and the time for a single immersion extraction was 72 h; the extracts were combined, and then the extracts were concentrated by a reduced pressure concentration method at 40° C. to obtain 3.5 kg of alcohol extract.

[0078] S2: The alcohol extract is subjected to a first silica gel column chromatography to obtain 12 components, which are recorded as Fr.1 to Fr.12, and the third component is collected and recorded as Fr.3; the fourth component is collected and recorded as Fr.4; the particle size of the silica gel used for the first silica gel column chromatography is 200 to 300 mesh; gradient elution is performed using petroleum ether-ethyl acetate with volume ratios of 98:2, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 25:75 and 0:100 and ethyl acetate-methanol with volume ratios of 90:10, 80:20, 50:50 and 0:100.

[0079] S3: The Fr.3 is subjected to a second silica gel column chromatography to obtain 6 components, which are recorded as Fr.3.1 to Fr.3.6. The first component is collected and recorded as Fr.3.1; the particle size of the silica gel used for the second silica gel column chromatography is 200 to 300 mesh; the second silica gel column chromatography adopts gradient elution with petroleum ether-ethyl acetate in volume ratios of 80:1, 60:1, 30:1, 15:1 and 0:100 respectively.

[0080] S4: Subject the Fr.3.1 to a first gel column chromatography and collect the second component, recorded as Fr.3.1.2; subject the Fr.3.1.2 to a second gel column chromatography and collect the second component, recorded as Fr.3.1.2.2; the eluent used in the first gel column chromatography is dichloromethane-methanol in a volume ratio of 1:1, and the eluent used in the second gel column chromatography is methanol.

[0081] S5: Fr.3.1.2.2 was separated by a first reverse phase column using 89 v / v% methanol-water as the eluent to obtain compound 1 (arnebidin).

[0082] S6: The Fr.4 is subjected to a third silica gel column chromatography to obtain 5 components, which are recorded as Fr.4.1 to Fr.4.5, and the second component is collected and recorded as Fr.4.2; the particle size of the silica gel used for the third silica gel column chromatography is 200 to 300 mesh, and the gradient elution is performed using dichloromethane-methanol with a volume ratio of 100:1, 80:1, 50:1, 25:1, 10:1 and 0:100.

[0083] S7: Subject Fr.4.2 to a third gel column chromatography, and collect the fourth component, which is recorded as Fr.4.2.4; subject Fr.4.2.4 to a fourth gel column chromatography, and collect the third component, which is recorded as Fr.4.2.4.3; the eluent used in the third gel column chromatography is dichloromethane-methanol in a volume ratio of 1:1, and the eluent used in the fourth gel column chromatography is methanol.

[0084] S8: Fr.4.2.4.3 was separated by a second reverse phase column using 87 v / v% methanol-water as the eluent to obtain compound 2 ((+)-(R)-de-O-methyllasiodiplodin).

[0085] The arnebidin prepared in Example 1 was subjected to NMR analysis and mass spectrometry analysis, and the results were as follows:

[0086] Molecular formula: C 32 H 26 O8; ESIMS: m / z 561[M+Na] + ; 1 H NMR(500MHz,CDCl3)δ:12.49(1H,s,8-OH),12.39(1H,s,5′-OH),12.30(1H,s,5-OH),12.08(1H,s,8′-OH),7.30(1H,d,J=9.3Hz,1H,H-7),7. 28(1H,d,J=9.3Hz,H-6),7.26(1H,d,J=9.3Hz,H-6′),7.24(1H,d,J=9.3Hz,H-7′),4.62(1H,dt,J=9.0,1.4Hz,H-13′),4.38(1H,dt,J=9.0,1 .4Hz,H-13),3.88(1H,td,J=4.5,1.2Hz,H-11′),3.82(1H,dd,J=9.0,4.5Hz,H-12),3.35(1H,ddd,J=9.0,4.5,2.6Hz,H-12′),2.68(1H,dd,J =2.6,1.2Hz,H-2),1.83(3H,d,J=1.4Hz,H3-15),1.72(3H,d,J=1.4Hz,H3-16′),1.61(3H,d,J=1.4Hz,H3-15′),1.54(3H,d,J=1.4Hz,H3-16); 13C NMR(126MHz, CDCl3)δ:195.5(C-4),193.1(C-1),182.6(C-4′),182.2(C-1′),159.0(C-5′),158.5(C-8′),157.3(C-8),15 6.3(C-5),144.3(C-2′),140.0(C-3′),138.6(C-14),137.8(C-14′),129.9(C-7′),129.3(C-6′),129.1(C-6),128.9(C-7) ,117.4(C-13′),116.8(C-13),112.7(C-10),112.4(C-9′),111.7(C-9),111.4(C-10′),49.2(C-3),45.3(C-2),40.6(C-12),38.1(C-11),37.6(C-12′),26.0(C-16),25.8(C-16′),18.7(C-15′),18.6(C-15). From the above test results, it can be seen that the compound prepared in Example 1 is arnebidin.

[0087] The (+)-(R)-de-O-methyllasiodiplodin prepared in Example 1 was subjected to NMR analysis, mass spectrometry analysis and specific rotation analysis. The results are as follows:

[0088] Molecular formula: C 16 H 22 O4; ESIMS: m / z 301[M+Na] + ,[α]20D=+20(c 0.2,MeOH), 1 HNMR(500MHz,CD3OD)δ:6.14(1H,d,J=2.5Hz,H-3),6.08(1H,d,J=2.5Hz,H-5),5.09(1H,m,H-13),3.19(1H,td,J=12.2,3.5Hz,H -6a),2.43(1H,m,H-6b),1.88(1H,m,H-12a),1.76(1H,m,H-12b),1.48(10H,m,H2-7,8,9,10,11,),1.29(3H,d,J=6.2Hz,H3-16); 13C NMR (126MHz, CD3OD)δ:173.1(C-14),165.9(C-1),163.5(C-3),149.7(C-5),111.8(C-4),106.0(C-15),101.8(C-2),75.9(C-13),34.2(C-6),32.1(C-12),32.0(C-7),28.2(C-8),25.6(C-11),25.4(C-10),22.5(C-9),20.4(C-16). From the above test results, it can be seen that the compound prepared in Example 1 is (+)-(R)-de-O-methyllasiodiplodin.

[0089] Test Example 1

[0090] Anti-triple negative breast cancer MDA-MB-231 cell viability assay:

[0091] Triple negative breast cancer MDA-MB-231 cells were placed in a 96-well plate at a density of 5000 cells per well. The cells were exposed to different concentrations of the test compound for 48 h. After the incubation, 10 μL of CCK-8 solution was added to each well and incubated at 37 °C for 2 h. The absorbance was then measured at 450 nm using a microplate reader (Molecular Devices, USA). Relative cell viability was calculated, and IC was calculated using GraphPadPrism 9.0.0 50 The results are listed in Table 1. Paclitaxel and β,β-dimethylacrylshikonin were used as positive control drugs.

[0092] Table 1 IC of inhibition of MDA-MB-231 cell activity 50 value

[0093] Test compound <![CDATA[IC 50 (μM)]]> arnebidin 1.16 (+)-(R)-de-O-methyllasiodiplodin 0.68 Paclitaxel 0.48 β,β-dimethylacrylshikonin 2.45

[0094] It can be seen that the compounds arnebidin and (+)-(R)-de-O-methyllasiodiplodin have good inhibitory activity on triple-negative breast cancer MDA-MB-231 cells.

[0095] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a phenolic compound and / or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating breast cancer, wherein the phenolic compound comprises compound 1 and / or compound 2:

2. The use according to claim 1, characterized in that: The breast cancer includes triple-negative breast cancer.

3. The use according to claim 1, characterized in that: The pharmaceutically acceptable salts include sodium salts or potassium salts.

4. The method for preparing a phenolic compound according to claim 1, characterized in that: The preparation method of the phenolic compound comprises the following steps: The root of Lithospermum erythrorhizon is extracted by using a low-grade alcohol aqueous solution to obtain an alcohol extract; The alcohol extract is subjected to a first silica gel column chromatography to collect the third and fourth components, which are respectively recorded as Fr.3 and Fr.4; the elution mode of the first silica gel column chromatography is gradient elution, and the eluents used in the gradient elution include petroleum ether-ethyl acetate and ethyl acetate-methanol in sequence; the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate is 98:2, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 25:75 and 0:100; the volume ratio of ethyl acetate to methanol in the ethyl acetate-methanol is 90:10, 80:20, 50:50 and 0:100 in sequence; The preparation method of compound 1 comprises the following steps: The Fr.3 is subjected to a second silica gel column chromatography, and the first component is collected and recorded as Fr.3.1; the eluent used in the second silica gel column chromatography is petroleum ether-ethyl acetate; the elution method of the second silica gel column chromatography is gradient elution, and the volume ratios of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate during the second silica gel column chromatography are 80:1, 60:1, 30:1, 15:1 and 0:100 respectively; The Fr.3.1 is subjected to a first gel column chromatography to collect the second component, which is recorded as Fr.3.1.2; the Fr.3.1.2 is subjected to a second gel column chromatography to collect the second component, which is recorded as Fr.3.1.2.2; the eluents used in the first gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the first gel column chromatography process is 20-80%; the eluent used in the second gel column chromatography is methanol; The Fr.3.1.2.2 is separated by a first reverse phase chromatographic column to obtain compound 1; the eluent used for separation by the first reverse phase chromatographic column is methanol-water; The preparation method of the compound 2 comprises the following steps: The Fr.4 is subjected to a third silica gel column chromatography, and the second component is collected and recorded as Fr.4.2; the eluents used in the third silica gel column chromatography are dichloromethane and methanol; the elution method of the third silica gel column chromatography is gradient elution, and the volume ratios of dichloromethane and methanol in the third silica gel column chromatography are 100:1, 80:1, 50:1, 25:1, 10:1 and 0:100 respectively; The Fr.4.2 is subjected to a third gel column chromatography to collect the fourth component, which is recorded as Fr.4.2.4; the Fr.4.2.4 is subjected to a fourth gel column chromatography to collect the third component, which is recorded as Fr.4.2.4.3; the eluents used in the third gel column chromatography are dichloromethane and methanol, and the volume fraction of dichloromethane in the third gel column chromatography process is 20-80%; the eluent used in the fourth gel column chromatography is methanol; The Fr.4.2.4.3 is subjected to a second reverse phase chromatography column separation to obtain compound 2; the eluent used for the second reverse phase chromatography column separation is methanol-water.

5. The use according to claim 4, characterized in that: The volume fraction of the lower alcohol in the lower alcohol aqueous solution is 50-100%; the lower alcohol includes methanol and / or ethanol; The volume ratio of the dry weight of the lithospermum root to the lower alcohol aqueous solution for single extraction is 4-6 kg: 6-7 L; The extraction temperature is 15-40° C., the number of extractions is 2-4 times, and the time for a single extraction is 24-120 hours.

6. The use according to claim 4, characterized in that: The gels used for the first gel column chromatography, the second gel column chromatography, the third gel column chromatography and the fourth gel column chromatography include hydroxypropyl dextran gel.

7. The use according to claim 4, characterized in that: The volume fraction of methanol in methanol-water used for separation by the first reverse phase chromatographic column is 85-95%; The volume fraction of methanol in methanol-water used for separation in the second reverse phase chromatography column is 86-90%.

8. A pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable excipient; the active ingredient is a phenolic compound and / or a pharmaceutically acceptable salt thereof, and the phenolic compound is the phenolic compound in the use according to any one of claims 1 to 7.

9. The pharmaceutical composition according to claim 8, characterized in that The content of the active component in the pharmaceutical composition is 0.1-99 wt%.

10. The pharmaceutical composition according to claim 8 or 9, characterized in that The dosage form of the pharmaceutical composition includes oral preparation or injection.

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