A compound derived from arnebia euchroma, a preparation method thereof and application thereof in anti-tumor

Abstract

This invention belongs to the pharmaceutical field and relates to a class of novel monoterpenoid phenol and benzoquinone derivatives from Xinjiang Lithospermum erythrorhizon, their preparation methods, and the application of these compounds in the preparation of antitumor drugs. In vitro antitumor activity experiments demonstrated that the compounds exhibited effective inhibitory activity against various tested tumor cell lines, particularly compound 1, which showed a significantly higher IC50 value against HCT-8 (human colon cancer cell line) and HepG2 (human liver cancer cell line). 50 The concentrations were 6.51 and 6.14 μM, respectively; the IC50 values ​​of compounds 2 and 3 against HepG2 were... 50 The concentrations were 4.27 and 3.45 μM, respectively; compound 4 showed IC50 values ​​against HCT-8, HGC-27 (human gastric cancer cell line), HepG2, and PC9 (human lung cancer cell line). 50 The concentrations were 5.08, 6.12, 2.31, and 5.32 μM, respectively; the IC50 values ​​of compound 5 against HGC-27 were... 50 The concentration was 5.69 μM. These results suggest that this type of compound could be used in the preparation of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of medicine, specifically relating to a class of novel monoterpenoid phenols and benzoquinones isolated from the traditional Chinese medicine Lithospermum erythrorhizon and their application in anti-tumor treatment. Background Technology

[0002] Cancer is a persistent public health challenge facing the world. As the second leading cause of death globally, the number of cancer cases and deaths is rising year by year, and cancer is projected to become the most significant obstacle to increasing life expectancy in the 21st century. The World Health Organization (WHO) estimates that the number of cancer cases worldwide may increase by 60% in the next 20 years, making prevention and control crucial. According to the latest cancer report on China published by the National Cancer Center in the *Journal of the National Cancer Center* in 2022, lung cancer, colorectal cancer, stomach cancer, liver cancer, and breast cancer in women are the five most common cancers, accounting for 57.4% of all new cases. Lung cancer, liver cancer, stomach cancer, colorectal cancer, and esophageal cancer are the five leading causes of cancer death, accounting for 69.3% of all deaths. Chemotherapy, as one of the most effective treatments for cancer, plays an important role in the treatment of metastatic, advanced-stage tumors and tumors with a tendency to spread throughout the body. Currently, many anticancer drugs are used in cancer treatment, such as 5-fluorouracil, cisplatin, etoposide, paclitaxel, and doxorubicin. However, these chemotherapy drugs have significant toxic side effects, such as nausea, vomiting, loss of appetite, decreased immunity, and oral ulcers. In addition, the frequent occurrence of tumor drug resistance makes it crucial to develop and research anti-tumor drugs with fewer toxic side effects, higher efficacy, and different mechanisms of action.

[0003] Natural compounds or their derivatives can prevent and treat tumors by targeting tumor marker molecules and activating related mechanisms. Natural compounds serve as important sources of lead compounds in anticancer drug development; for example, camptothecin, vincristine, and gentian violet toxin from traditional Chinese medicine exhibit excellent anticancer activity and are widely used clinically. Our project team has been dedicated to discovering the pharmacodynamic material basis and active lead compounds of commonly used traditional Chinese medicines. Combining clinical applications, literature reviews, and laboratory screening for antitumor activity, we discovered that the extract of *Lithospermum erythrorhizon* from Xinjiang possesses good antitumor activity.

[0004] Xinjiang Lithospermum erythrorhizon is the dried root of Arnebiaeuchroma (Royle) Johnst., a plant belonging to the genus Arnebia in the family Boraginaceae. It is the main source of the traditional Chinese medicine Lithospermum erythrorhizon, first recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica). It is mainly distributed in Xinjiang, Tibet, Inner Mongolia, and Gansu provinces. It has a bitter taste and cold properties, and is used to clear heat and cool the blood, promote blood circulation and detoxify, and promote rash eruption and eliminate rashes. It is primarily used to treat conditions such as blood heat and toxicity, purplish-black rashes, incomplete measles eruption, sores, eczema, and burns. Chemical composition and pharmacological activity studies have shown that Xinjiang Lithospermum erythrorhizon contains naphthoquinones, monoterpenes (phenols and benzoquinones), triterpenes and steroids, phenolic acids and organic salts, aliphatic compounds and esters, exhibiting antitumor, antibacterial, anti-inflammatory, antiviral, antioxidant, hepatoprotective, and immunomodulatory pharmacological activities. The most reported compounds are naphthoquinones and monoterpenoid phenols and benzoquinones, with naphthoquinones considered to be the main active components of Lithospermum erythrorhizon. Current research on the activity and related mechanisms of shikonin and benzoquinones is relatively in-depth, but these compounds have a relatively simple parent nucleus (5,8-dihydroxy-2-isohexene-1,4-naphthoquinone). Monoterpenoid phenols and benzoquinones, on the other hand, have diverse parent nuclei and have shown good activity in cytotoxicity and antiviral activity. Summary of the Invention

[0005] The applicant isolated a class of novel monoterpenoid phenols and benzoquinones (1-5) from *Lithospermum erythrorhizon*, with the chemical structures shown below. These structures have a 6 / 10 / 5 tricyclic core or a 6 / 6 / 5 tricyclic benzogeijerene skeleton, with one or two methoxy substitutions on the hydroquinone or hydroquinone ring, and the five-membered ring is an α,β-unsaturated cyclobutyrolactone or a cyclobutyrolactone with a double bond substitution at the α-position. In in vitro antitumor activity evaluation, these compounds exhibited significant cytotoxic activity against human colon cancer cell line (HCT-8), human pancreatic cancer cell line (PANC-1), human gastric cancer cell line (HGC-27), human liver cancer cell line (HepG2), and human lung cancer cell line (PC9). Currently, there are no reports of isolating the above-mentioned monoterpenoid phenols and benzoquinones from the traditional Chinese medicine *Lithospermum erythrorhizon*, nor are there any patent documents on the preparation methods and antitumor activities of the above monomers. The technical problem to be solved by the present invention is to provide new monoterpenoid phenols and benzoquinones, their pharmaceutically acceptable salts, pharmaceutical compositions thereof, and their use in the preparation of antitumor drugs.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0007] The first aspect of the present invention is to provide a class of novel compounds 1-5 (1: 3,4-dimethoxy-(+)-arnebinol A; 2:(+)-arnebinone E; 3:(-)-arnebinone E; 4:3,4-dimethoxyarnebinol C; 5:arnebinone C) derived from Xinjiang Lithospermum erythrorhizon and pharmaceutically acceptable salts of compounds 1 and 4, the structures of which are as follows:

[0008]

[0009] The second aspect of this invention provides a method for preparing novel monoterpenoid phenols and benzoquinones from Xinjiang Lithospermum erythrorhizon, characterized by the following steps: 95% and 80% ethanol reflux extraction of Xinjiang Lithospermum erythrorhizon, followed by extraction of the concentrated extract using petroleum ether, ethyl acetate, and water-saturated n-butanol. The ethyl acetate fraction is purified by silica gel column solid-phase extraction, silica gel column chromatography, Sephadex LH-20 gel column chromatography, and preparative HPLC to obtain compounds 1-5. Their structures are identified by spectroscopic methods such as UV, IR, NMR, MS, ECD, and X-ray single-crystal diffraction, revealing them to be novel monoterpenoid phenols and benzoquinones, substances not previously reported.

[0010] A third aspect of the present invention is to provide a pharmaceutical composition containing an effective dose of the compound described in the first aspect or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Typically, the pharmaceutical composition of the present invention contains 0.1 to 95% by weight of the compound of the present invention. In unit dosage forms, the content of the compound of the present invention is generally 0.1 to 100 mg, and preferably 4 to 50 mg.

[0011] Pharmaceutical compositions of the compounds of the present invention can be prepared according to methods known in the art. For this purpose, if desired, the compounds of the present invention can be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to formulate suitable administration or dosage forms for use as human or veterinary medicine.

[0012] The compounds of this invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum, or rectum.

[0013] The compounds of this invention or pharmaceutical compositions containing them can be administered via injection. Injection includes intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, and acupoint injection, etc.

[0014] Dosage forms can be liquid or solid. Liquid dosage forms include true solutions, colloids, microparticles, emulsions, and suspensions. Other dosage forms include tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, and lyophilized powder injections.

[0015] The compounds of this invention can be formulated into conventional formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0016] For example, various carriers known in the art can be widely used to formulate unit-dose dosage forms into tablets. Examples of carriers include diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, and aluminum silicate; humectants and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, shellac, methylcellulose, potassium phosphate, and polyvinylpyrrolidone; disintegrants such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, methylcellulose, and ethylcellulose; disintegration inhibitors such as sucrose, tristearate, cocoa butter, and hydrogenated oil; absorption promoters such as quaternary ammonium salts and sodium dodecyl sulfate; and lubricants such as talc, silica, corn starch, stearates, boric acid, liquid paraffin, and polyethylene glycol. Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0017] For example, various carriers known in the art can be widely used to formulate the drug delivery unit into pills. Examples of carriers include diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, glyceryl monostearate, kaolin, talc, etc.; binders such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste, or flour paste, etc.; and disintegrants such as agar powder, dried starch, alginate, sodium dodecyl sulfate, methylcellulose, ethylcellulose, etc.

[0018] For example, to formulate the drug delivery unit into a capsule, the active ingredient, the compound of the present invention, is mixed with the various carriers described above, and the resulting mixture is placed in a hard gelatin capsule or a soft capsule. Alternatively, the active ingredient, the compound of the present invention, can be formulated as a microcapsule, suspended in an aqueous medium to form a suspension, or filled into a hard capsule or formulated as an injectable preparation for use.

[0019] For example, the compounds of this invention can be formulated into injectable formulations, such as solutions, suspension solutions, emulsions, and lyophilized powders for injection. These formulations can be aqueous or non-aqueous and may contain one or more pharmacodynamically acceptable carriers, diluents, binders, lubricants, preservatives, surfactants, or dispersants. Diluents may be selected from water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxyethylene isostearyl alcohol, polyoxyethylene sorbitan esters, fatty acid esters, etc. Furthermore, to prepare isotonic injections, appropriate amounts of sodium chloride, glucose, or glycerol can be added to the injectable formulations. In addition, conventional solubilizers, buffers, pH adjusters, etc., may also be added. These excipients are commonly used in the art.

[0020] In addition, colorants, preservatives, flavorings, tasters, sweeteners or other materials may be added to pharmaceutical preparations if necessary.

[0021] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0022] The dosage of the pharmaceutical composition of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, personality and individual response of the patient or animal, the route of administration, the frequency of administration, and the therapeutic purpose. Therefore, the therapeutic dosage of the present invention can vary widely. Generally speaking, the dosage of the pharmaceutical components used in the present invention is well known to those skilled in the art. The actual amount of drug contained in the final formulation of the compound composition of the present invention can be appropriately adjusted to achieve the required therapeutic dose and fulfill the preventive or therapeutic purpose of the present invention. The suitable daily dosage range of the compound of the present invention is: 0.001–100 mg / kg body weight, preferably 0.1–60 mg / kg body weight, more preferably 1–30 mg / kg body weight, and most preferably 2–15 mg / kg body weight. For adult patients, the daily dosage of the compound of the present invention is 10–500 mg, preferably 20–100 mg, which can be taken once or divided into 2–3 doses; for children, the dosage is 5–30 mg / kg body weight, preferably 10–20 mg / kg body weight. The above dosage can be administered as a single dose or divided into several doses, such as two, three, or four doses, depending on the clinical experience of the attending physician and the dosing regimen of the treatment. The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs.

[0023] The fourth aspect of this invention is to provide the use of the compound described in the first aspect or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition described in the third aspect, in the preparation of an antitumor drug. The in vitro cytotoxic activity of compounds 1-5 against human colon cancer cell lines (HCT-8), human pancreatic cancer cell line (PANC-1), human gastric cancer cell line (HGC-27), human liver cancer cell line (HepG2), and human lung cancer cell line (PC9) was evaluated using the MTT assay. The above experiments were conducted using IC50 assay. 50 <20 μM was considered an effective standard; the results confirmed that the above compounds had inhibitory effects on various tumor cell lines, especially compound 1, which had an IC50 value of <20 μM for HCT-8 and HepG2. 50 The concentrations were 6.51 and 6.14 μM, respectively; the IC50 values ​​of compounds 2 and 3 against HepG2 were... 50 The concentrations were 4.27 and 3.45 μM, respectively; the IC50 values ​​of compound 4 for HCT-8, HGC-27, HepG2, and PC9 were 4.27 and 3.45 μM, respectively. 50 The concentrations were 5.08, 6.12, 2.31, and 5.32 μM, respectively; the IC50 values ​​of compound 5 against HGC-27 were... 50 It is 5.69 μM.

[0024] Beneficial technical effects:

[0025] 1. The novel compounds of monoterpenoid phenol and benzoquinone from Xinjiang Lithospermum erythrorhizon of the present invention have significant in vitro cytotoxic activity.

[0026] 2. The novel compounds of monoterpenoid phenol and benzoquinone from Xinjiang Lithospermum erythrorhizon of this invention have novel structures and have not been reported in patent literature, and have the potential to be further developed into anti-tumor drugs. Detailed Implementation

[0027] The following examples and pharmacological activity experiments are used to further illustrate the present invention, but they do not imply any limitation on the present invention.

[0028] Example 1: Preparation method of monomeric compounds 1-5 from Lithospermum erythrorhizon, Xinjiang

[0029] 50 kg of dried roots of Lithospermum erythrorhizon from Xinjiang were pulverized and extracted twice with 95% ethanol under reflux for 2 hours each time. After filtration, the supernatant was collected, and the residue was extracted again with 80% ethanol for 2 hours. The three extracts were combined, and the solvent was recovered under reduced pressure to obtain approximately 4 kg of total ethanol extract ZCT. The total extract was dispersed in 15 L of water and then extracted three times with 15 L of petroleum ether, ethyl acetate, and water-saturated n-butanol, respectively. The extract was concentrated under reduced pressure to obtain 1.7 kg of ethyl acetate fraction (ZCEA). The ethyl acetate fraction ZCEA was subjected to solid-phase extraction (d = 13 cm, h = 80 cm) after mixing with 2.5 kg of 100-200 mesh silica gel. The elution ratios of petroleum ether:ethyl acetate were 100:1, 80:1, 50:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, and 0:100 to obtain ten subfractions, which were named ZCEA-A to ZCEA-J.

[0030] The ZCEA-G fraction (approximately 120g) was mixed with 250g of 100-200 mesh silica gel and subjected to 100-200 mesh silica gel column chromatography (d=14cm, h=106cm). Elution was performed using petroleum ether:ethyl acetate at ratios of 5:1, 3:1, 2:1, 1:1, 1:2, and 0:100. The fractions were concentrated to obtain 344 fractions from Fra.G-1 to Fra.G-344. Fra.G-94 to Fra.G-118 were combined and subjected to Sephadex LH-20 gel column chromatography (petroleum ether:dichloromethane = 5:1 to 1:1 gradient elution), designated as column G5, yielding 63 fractions from G5.1 to G5.63. HPLC analysis of fractions G5.22 to G5.46 revealed three peaks with identical retention times and UV values. Fractions G5.33 to G5.36 were analyzed using C0.05... 18 Reversed-phase semi-preparative liquid chromatography (MeCN / H2O, 55:45, v / v, protected from light) was used to purify compound A (24.3 mg) and 5 (11.9 mg). Compound A was then semi-preparatively resolved using a Chiralpak AD-RH (250×10 mm, 5 μm, MeCN / H2O, 70:30, v / v) chiral column to obtain compounds 2 (10.5 mg) and 3 (4 mg). The G5 column was flushed with methanol, and the concentrated sample was subjected to secondary Sephadex LH-20 gel chromatography (petroleum ether:dichloromethane:methanol = 5:5:1 and 5:5:2 gradient elution), designated as column G5A, yielding 61 fractions from G5A.1 to G5A.61. G5A.24 was further purified by C... 18Compounds 1 (20.6 mg) and 4 (9.8 mg) were purified twice by reversed-phase semi-preparative liquid chromatography (1: MeCN / H2O, 55:45, v / v; 2: MeCN / H2O, 50:50, v / v). Their structures were identified by UV, IR, NMR, MS, ECD, and X-ray single-crystal diffraction. They are novel monoterpenoid phenols and benzoquinones with a 6 / 10 / 5 tricyclic core or a 6 / 6 / 5 tricyclic benzogeijerene skeleton, one or two methoxy groups on the hydroquinone or hydroquinone ring, and the five-membered ring being α,β-unsaturated cyclobutyrolactone or cyclobutyrolactone with a double bond at the α-position.

[0031] The spectral information and NMR signal assignments of the above-mentioned new compounds are as follows:

[0032] Compound 1

[0033]

[0034] 3,4-dimethoxy-(+)-arnebinol A: White amorphous powder; specific rotation +153.8(c 0.1,MeOH); UV(MeOH)λ max (logε)299(2.33),222sh(2.89),203(3.37)nm; ECD(c 0.25mg / mL,MeOH) λ max (Δε)259(+3.01),224(-34.11)nm; IRν max 3384,2963,2938,2871,1727,1647,1597,1495, 1450,1419,1362,1326,1288,1236,1206,1175,1120,1042,1004,987,954,836cm -1 HRESIMS m / z 317.1380[M+H] + (calcd for C 18 H 21 O5,317.1383).

[0035] 1 H NMR (500MHz, CD3OD), δ H:6.59(1H,s,H-2),6.70(1H,t,J=1.5Hz,H-6),6.96(1H, d,J=1.5Hz,H-7),1.91(1H,m,H-9β),2.74(1H,m,H-9α),2.21(1H,m,H-10β),1.89(1H,m ,H-10α),5.10(1H,t,J=8.0Hz,H-11),3.33(1H,overlap,H-13β),2.37(1H,d,J=15.0Hz, H-13α),1.58(3H,s,H3-16),3.83(3H,s,CH3O-3),3.77(3H,s,CH3O-4).

[0036] 13 C NMR (125MHz, CD3OD), δ C :154.5(C-1),102.6(C-2),152.8(C-3),143.0(C-4),125.6 (C-5),78.8(C-6),152.0(C-7),129.8(C-8),26.2(C-9),25.5(C-10),121.5(C-11),142.3( C-12),26.6(C-13),118.1(C-14),177.5(C-15),22.2(C-16),56.2(CH3O-3),62.5(CH3O-4).

[0037] Single-crystal X-ray diffraction data: C 55 H 64 O 16 ,[3M+CH3OH]=981.06,0.21×0.18×0.03mm 3 ,orthorhombic, α=90°, β=90°, γ=90°, T=116.2(2)K, space group P212121(no.19), Z=4, μ(Cu Kα)=0.790mm -1 , wR(F 2 =0.1181. Flack parameter =0.00(10). Single crystal data is stored in the Cambridge Crystal Database Centre (CCDC No.: 2113159).

[0038] Compound 2

[0039]

[0040] (+)-arnebinone E: Yellow amorphous powder; specific rotation +23.6 (c 0.10, MeCN); UV (MeCN) λ max (logε) 269 (3.13) nm; ECD (c 0.25 mg / mL, MeCN) λ max (Δε) 371 (+1.98), 291 (+3.88), 255 (-9.66), 219 (-30.12) nm; IR ν max 3101, 2958, 2932, 1736, 1680, 1640, 1604, 1456, 1346, 1318, 1211, 1108, 1052, 976 cm -1 ; HRESIMS m / z 301.1068 [M+H] + (calcd for C 17 H 17 O5, 301.1070).

[0041] 1 1H NMR (500 MHz, CDCl3), δ H : 6.07 (1H, s, H-2), 6.61 (1H, d, J = 1.5 Hz, H-6), 6.72 (lH, d, J = 1.5 Hz, H-7), 1.90 (1H, m, H-9β), 2.84 (1H, m, H-9α), 2.29 (1H, m, H-10β), 1.90 (1H, m, H-10α), 5.16 (1H, t, J = 7.5 Hz, H-11), 3.40 (1H, d, J = 13.5 Hz, H-13β), 2.38 (1H, d, J = 13.5 Hz, H-13α), 1.50 (3H, s, H3-16), 3.87 (3H, s, CH3O-3).

[0042] 13 13C NMR (125 MHz, CDCl3), δ C : 186.0 (C-1), 108.0 (C-2), 158.0 (C-3), 180.6 (C-4), 134.1 (C-5), 73.6 (C-6), 147.6 (C-7), 129.8 (C-8), 25.5 (C-9), 24.7 (C-10), 122.9 (C-11), 138.2 (C-12), 25.6 (C-13), 148.8 (C-14), 174.3 (C-15), 23.4 (C-16), 56.7 (CH3O-3).

[0043] Single crystal X-ray diffraction data: C 17 H 16O5,M=300.30,0.31×0.22×0.05mm 3 monoclinic α=90°, β=94.4777(12)°, γ=90°, T=99.99(10)K,space group P21(no.4),Z=2,μ(Cu Kα)=0.846mm -1 ,wR(F 2 =0.0825. Flack parameter = -0.06(10). Single crystal data is stored in the Cambridge Crystal Database Centre (CCDC No.: 2093858).

[0044] Compound 3

[0045]

[0046] (-)-arnebinone E: Yellow amorphous powder; specific rotation -24.0(c 0.10,MeCN); ECD(c0.25 mg / mL,MeCN)λ max (Δε) 372(-1.79), 291(-3.37), 256(+8.90), 219(+27.22) nm; UV, IR, HRESIMS and NMR data are consistent with those of compound 2.

[0047] Single-crystal X-ray diffraction data: C 17 H 16 O5,M=300.30,0.31×0.24×0.02mm 3 monoclinic α=90°, β=94.788°(5), γ=90°, T=116.55(10)K,space group P21(no.4),Z=2,μ(Cu Kα)=0.839mm -1 ,wR(F 2 =0.0972. Flack parameter =0.1(2). Single crystal data is stored in the Cambridge Crystal Database Centre (CCDC No.: 2150404).

[0048] Compound 4

[0049]

[0050] 3,4-dimethoxyarnebinol C: White amorphous powder; specific rotation +157.0(c 0.1,MeOH); UV (MeOH)λ max (logε)297(2.58),225sh(3.13),202(3.60)nm; ECD(c 0.1mg / mL,MeOH)λ max (Δε)304(+0.80),260(-2.05),221(+15.70)nm; IRν max 3419,2963,2940,2841,1741,1600, 1504,1457,1425,1362,1338,1272,1240,1156,1127,1046,987,958,926,827cm -1 HRESIMS m / z 317.1381 [M+H] + (calcd for C 18 H 21 O5,317.1384).

[0051] 1 HNMR (500MHz, CD3OD), δ H :6.58(1H,s,H-2),5.61(1H,d,J=6.0Hz,H-5),3.15(1H,dt,J=6.0,1.5Hz,H-6),2.39(1H,d,J=16.5Hz,H-8β),2.90(1H ,d,J=16.5Hz,H-8α),1.20(3H,s,H3-9),5.71(1H,dd,J=18.0,10.5Hz,H-10),4.92(1H,dd,J=18.0,1.0Hz,H-11a),4.93 (1H,dd,J=10.5,1.0Hz,H-11b),5.82(1H,d,J=1.5Hz,H-13a),6.24(1H,d,J=1.5Hz,H-13b),3.82(3H,s,CH3O-3),3.83(3H,s,CH3O-4).

[0052] 13 C NMR (125MHz, CD3OD), δ C:152.1(C-1),102.5(C-2),152.5(C-3),142.7(C-4),126.5 (C-4a),75.5(C-5),50.1(C-6),38.6(C-7),34.1(C-8),115.6(C-8a),24.6(C-9),141.9(C- 10),115.1(C-11),139.6(C-12),123.6(C-13),173.0(C-14),56.2(CH3O-3),61.8(CH3O-4).

[0053] Single-crystal X-ray diffraction data: C 18 H 20 O5,M=316.344,0.38×0.17×0.11mm 3 monoclinic α=90°, β=97.163°(6), γ=90°, T=114.45(10)K,space group P21(no.4),Z=2,μ(Cu Kα)=0.798mm -1 ,wR(F 2 = 0.0879. Flack parameter = 0.07(11). Single crystal data is stored in the Cambridge Crystal Database Centre (CCDC No.: 2144856).

[0054] Compound 5

[0055]

[0056] Arnebinone C: Yellow amorphous powder; +34.9(c 0.06,MeCN); UV(MeCN)λ max (logε)265 (2.99)nm; ECD (c 0.29mg / mL, MeCN)λ max (Δε)287(+0.35),259(-0.67),221(+2.51)nm; IR ν max 3018,2973,2941,2840,1765,1677,1654,1639,1609,1462,1444,1356,1311,1228,1148, 1123,1027,970,755cm -1 HRESIMS m / z 301.1067[M+H] + (calcd for C17 H 17 O5,301.1070).

[0057] 1 H NMR (500MHz, CDCl3), δ H :6.00(1H,s,H-2),5.44(1H,d,J=6.5Hz,H-5),3.09(1H, dt,J=6.5,2.0Hz,H-6),2.37(1H,d,J=19.0Hz,H-8β),2.86(1H,d,J=19.0Hz,H-8α),1.20 (3H,s,H3-9),5.75(1H,dd,J=17.5,11.0Hz,H-10),4.99(1H,d,J=17.5Hz,H-11a),5.11(1H,d, J=11.0Hz,H-11b),5.84(1H,d,J=2.0Hz,H-13a),6.35(1H,d,J=2.0Hz,H-13b),3.85(3H,s,CH3O-3).

[0058] 13 C NMR (125MHz, CDCl3), δ C :186.1(C-1),107.6(C-2),159.1(C-3),179.8(C-4),134.2 (C-4a),70.9(C-5),47.7(C-6),37.9(C-7),31.7(C-8),144.2(C-8a),24.9(C-9),13 9.8(C-10),115.8(C-11),135.6(C-12),124.6(C-13),169.5(C-14),56.6(CH3O-3).

[0059] Experimental Example 1: In vitro cytotoxicity experiment of monomeric compounds from Xinjiang Lithospermum erythrorhizon.

[0060] Experimental materials: Monomer compounds 1–5 isolated from Lithospermum erythrorhizon in Xinjiang, prepared with DMSO at an initial concentration of 1.0 × 10⁻⁵. -2 mol·L -1 Dilute as needed before administration. DMSO: Sigma-Aldrich, USA; MTT (tetramethylazophos) cell viability assay kit: Sigma-Aldrich, USA; DMEM and 1640 medium, fetal bovine serum (FBS) and phosphate buffer: HyClone, USA; trypsin-EDTA digestion solution: Jiangsu Kaiji Biotechnology Co., Ltd.; human tumor cell lines HCT-8, PANC-1, HGC-27, HepG2 and PC9: Cell Bank of Chinese Academy of Sciences (Shanghai).

[0061] Groups: (1) Control group: 10% FBS medium; (2) Sample group: 10% FBS medium with monomeric compound added; (3) Positive control group: 10% FBS medium with paclitaxel added.

[0062] Cell culture: Cells were passaged in DMEM or 1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 mg / L streptomycin at 37°C in a 5% CO2 incubator. Logarithmic growth phase cells were selected for the experiment.

[0063] MTT assay: Logarithmic growth phase cells were taken, digested, and thoroughly pipetted into a single-cell suspension. Cells were counted using a hemocytometer and then diluted to 1×10⁻⁶. 4 The sample was inoculated at 100 μL / well in a 96-well plate. Five different concentration groups were set up for each sample. Then, 100 μL of culture medium for each concentration level was added to the experimental wells, with three replicates for each concentration. An equal volume of solvent was added to the control group. After incubating the 96-well plate at 37°C and 5% CO2 for 72 hours, the culture medium was discarded. 50 μL of MTT (2 mg / mL) was added to each well, and the plate was incubated at 37°C for another 2 hours. The supernatant was then removed, and 150 μL of DMSO was added to each well to dissolve the formazan precipitate. The plate was then shaken for 10 minutes to ensure complete dissolution. The absorbance at 570 nm was measured using a Thermo MK3 microplate reader. 570 Using solvent-treated tumor cells as the solvent control group, the inhibition rate of tumor cell growth was calculated according to the following formula: Tumor cell growth inhibition rate (%) = (1-A) / ( ... 570 Group A 570 The solvent control group was multiplied by 100%. A dose-response curve was then obtained by plotting the drug concentration against the tumor cell growth inhibition rate, and the half-maximal inhibitory concentration (IC50) of the drug was read from the curve. 50 )value.

[0064] IC 50 <20 μM was the effective standard. The results showed that compounds 1-5 had effective inhibitory activity against all tested tumor cell lines (Table 1), especially compound 1, which had an IC50 value against HCT-8 and HepG2. 50 The concentrations were 6.51 and 6.14 μM, respectively; the IC50 values ​​of compounds 2 and 3 against HepG2 were... 50 The concentrations were 4.27 and 3.45 μM, respectively; the IC50 values ​​of compound 4 for HCT-8, HGC-27, HepG2, and PC9 were 4.27 and 3.45 μM, respectively. 50 The concentrations were 5.08, 6.12, 2.31, and 5.32 μM, respectively; the IC50 values ​​of compound 5 against HGC-27 were... 50 It is 5.69 μM.

[0065] Experimental results show that the monoterpenoid phenols and benzoquinone compounds 1-5 from Xinjiang Lithospermum erythrorhizon described in this invention have significant cytotoxic activity and can be used to prepare antitumor drugs.

[0066] Table 1. Cytotoxic activity of monomeric compounds against tumor cells

[0067]

[0068] .

Claims

1. A compound represented by the following structural formula or pharmaceutically acceptable salts of compounds 1 and 4: 。 2. A pharmaceutical composition, characterized by, A pharmaceutical composition comprising, as an effective ingredient, a compound according to claim 1 or pharmaceutically acceptable salts of compounds 1, 4 and a pharmaceutically acceptable carrier thereof.

3. Pharmaceutical composition according to claim 2, characterized in that The dosage form of the pharmaceutical composition is selected from the group consisting of tablets, capsules, pills, granules, oral solutions or suspensions.

4. Use of the compound according to claim 1 or pharmaceutically acceptable salts of compounds 1 and 4 for the preparation of an antitumor agent.

5. Use according to claim 4, characterized in that, The tumor is colon cancer, pancreatic cancer, gastric cancer, liver cancer and lung cancer.

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