Application of myristica fragrans glycoside compound in preparation of antitumor drugs

Abstract

The invention relates to an application of a nutmeg glycoside compound in preparation of antitumor drugs. The nutmeg glycoside compound has at least one compound represented by the following structure. And # imgabs0. The compound has a relatively good anti-tumor effect, especially an anti-liver cancer effect.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of a nutmeg glycoside compound in the preparation of anti-tumor drugs, especially in the application of anti-liver cancer activity. Background Art

[0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in humans, with a long-term high incidence and mortality rate. At present, there is still a lack of effective clinical means to treat liver cancer, and more than 90% of malignant solid tumors are difficult to achieve satisfactory therapeutic effects. Cytotoxic compounds have always been the main body of cancer treatment drugs, but they have poor efficacy on solid tumors, have large toxic side effects and are prone to drug resistance. Drugs are equivalent to bricks and stones for building buildings. They are compounds derived from nature, especially small molecule compounds, which are produced by organisms to resist natural enemies and adapt to the environment. They have diverse structures and a wide range of physiological activities, and their toxic side effects are relatively small compared to synthetic compounds.

[0003] The four compounds in this application are all derived from the seeds of Myristica fragrans Houtt., a plant of the genus Myristica in the family Myristicaceae. It is a medicine and food of the same origin. The 2020 edition of the "Chinese Pharmacopoeia" records that nutmeg has the effects of warming the middle and promoting qi, astringing the intestines and stopping diarrhea; it can be used for spleen and stomach deficiency and cold, chronic diarrhea, abdominal distension and pain, and vomiting due to poor appetite. Nutmeg contains phenylpropanoid carcinogenic chemical components such as myristicin, safrole, eugenol, and elegans, but there are no reports of carcinogenicity caused by nutmeg in clinical practice or as food. The above-mentioned chemical components are also chemical defense substances of nutmeg, and they are also autotoxic to the plant itself. The survival of plants is the result of long-term natural selection. In order to reduce their own toxicity, plants will also produce some "detoxification" components. One of the ways is to make them into glycosides. Glycoside compounds contain both cytotoxic aglycones and sugar groups that have a protective effect on normal human cells. The four compounds in the present invention all belong to phenylpropanoid glycosides. After screening by the MTT method, they have an inhibitory effect on HepG2 and SNU-739 liver cancer cells. Summary of the invention

[0004] The purpose of the present invention is to overcome the defects of the prior art and provide a use of a nutmeg glycoside compound in the preparation of an anti-tumor drug, wherein the compound has good anti-tumor effect, especially anti-liver cancer effect.

[0005] To achieve the above object, the present invention adopts the following technical solution:

[0006] Application of a nutmeg glycoside compound in the preparation of anti-tumor drugs.

[0007] The above application, wherein the myristica fragrans glycoside compound has at least one compound shown in the following structure;

[0008]

[0009] Furthermore, the myristica fragrans glycoside compound is a mixture of compounds 1, 2, 3 and 4 in any proportion.

[0010] Furthermore, the application of nutmeg glycoside compounds in the preparation of anti-liver cancer drugs.

[0011] The structures of compounds 1-3 mainly contain allylphenylpropanoids and single-chain disaccharide fragments, with rhamnose on the outside and linked to the 6th position of the inner glucose. Compound 4 contains a monosaccharide chain β-D-pyranose glucopyranose-β-D-pyranose glucopyranose fragment. The chemical names of compounds 1 to 4 are as follows (structural formulas are as follows Figure 1 shown):

[0012] Compound 1: 1-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranosyl)-2-hydroxy-4-allylbenzene;

[0013] Compound 2: 2-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranosyl)-1-hydroxy-4-allylbenzene;

[0014] Compound 3: 2-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranosyl)-1-hydroxy-6-methoxy-4-allylbenzene;

[0015] Compound 4: [4-(3-hydroxy-1-trans-propenyl)-2,6-dimethoxyphenyl]-6-O-β-D-pyranoglucopyranosyl-β-D-pyranoglucoside.

[0016] Traditional Chinese medicine has the characteristics of multiple components and multiple targets. The effective parts of the above four compound combinations, especially the combination of compounds 1-3, will also have good anti-tumor activity.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention adopts MTT method, takes doxorubicin as positive control, screens the inhibitory effect of the above four compounds on liver cancer cells HepG2 and SNU-739, and their IC 50 The IC values ​​for SNU-739 cells were: 21.92±0.48μM, 26.23±1.06μM, 50.52±1.87μM, 40.68±1.49μM respectively. 50The concentrations of nutmeg glycoside compounds 1, 2, 3 and 4 are respectively 28.19±0.13μM, 32.72±0.56μM, 51.85±2.33μM and 96.37±1.27μM. The experimental results show that the nutmeg glycoside compounds 1, 2, 3 and 4 have good anti-tumor effects, especially anti-liver cancer effects, and can be used to prepare anti-tumor drugs, especially anti-liver cancer drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Chemical structures of four compounds;

[0020] Figure 2 The results of the activity assay of compound 1-4 (128 μM) on HepG2 and SNU-739. DETAILED DESCRIPTION

[0021] The technical solution of the present invention is further described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0022] The four glycoside compounds of nutmeg in the present application are all known compounds, which can be extracted and obtained by referring to the method in Example 1, or prepared by referring to the method in existing literature. The relevant existing literature information is as follows.

[0023] Compounds 1 and 2: Ly TN, Yamauchi R, Shimoyamada M, et al. Isolation and structural elucidation of some glycosides from the rhizomes of smaller galanga (Alpinia officinarum Hance) [J]. J Agric Food Chem, 2002, 50(17): 4919-4924.

[0024] Compound 3: Wang XY, Xu M, Yang CR, et al. Phenylpropanoid glycosides from the seeds of Michelia hedyosperma[J]. Food Chemistry, 2011, 126(3): 1039-1043.

[0025] Compound 4: Jin C, Micetich RG, Daneshtalab M. Phenylpropanoid glycosides from Stellera chamaejasme[J]. Phytochemistry, 1999, 50(4): 677-680.

[0026] Example 1

[0027] The present invention relates to the application of four glycoside compounds in nutmeg in anti-liver cancer. To achieve the purpose, the following methods are mainly adopted:

[0028] Take 6.0kg of dried nutmeg kernels, crush them, soak them in 3 times the weight of methanol for 24 hours, reflux and extract 3 times, 1 hour each time, collect the filtrate, and concentrate under reduced pressure to recover methanol to obtain the extract. After extraction with petroleum ether and n-butanol respectively, take the n-butanol layer and pass it through a silica gel column, and use the eluent CHCl 3 :CH 3 OH:H 2 O (7:3:0.5, v / v) for elution and separation, and the eluate was collected; then the eluate was separated by MCI column (eluent CH 3 OH:H 2 (0, 30:70, v / v) elution separation, C 18 Column (eluent CH 3 OH:H 2 The four glycosides can be prepared by eluting and separating with 4% paraformaldehyde (2:4, 35:65, v / v) and collecting single components for TLC detection.

[0029] The structures of the four glycoside compounds were determined by MS, 1 H and 13 C NMR spectrum data analysis, its NMR data are shown in Tables 1 and 2, and its chemical structure is deduced as follows Figure 1 As shown, they are consistent with the data reported in the literature by Ly TN, Wang XY and Niwa M respectively.

[0030] Table 1 Compounds 1, 2, and 3 1 H NMR (500 MHz, DMSO-d 6 )and 13 C NMR (125 MHz, DMSO-d 6 )data

[0031]

[0032]

[0033] Table 2 Compound 4 1 H NMR (500 MHz, DMSO-d 6 )and 13 C NMR (125 MHz, DMSO-d 6 )data

[0034]

[0035]

[0036] Application test

[0037] The MTT method was used to select HepG2 and SNU-739 human liver cancer cells in the logarithmic growth phase, digested with trypsin, centrifuged at 1000 rpm for 5 min, discarded the supernatant, added DMEM medium to dilute, and concentrated to 5×10 3 Cells were seeded into 96-well plates and incubated at 37°C with 5% CO 2 Incubate in an incubator for 24 h. Doxorubicin (DOX) was used as a positive control, and the half-maximal inhibitory concentration of doxorubicin on HepG2 was measured as IC 50 =1.17±0.09μM, the half inhibitory concentration for SNU-739 is IC 50 =1.53±0.08μM. The blank group only had DMEM medium and no drug treatment. Compounds 1-4 were prepared into uniform solutions with concentrations of 2μM, 4μM, 8μM, 16μM, 32μM, 64μM, and 128μM, respectively. 100μL was added to each well of the experimental group. Three parallel wells were set up and repeated three times independently. After cell administration, the cells were incubated at 37°C and 5% CO 2 Incubate in the incubator for 48 hours, then add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours. Terminate the culture, carefully remove the culture medium, add 100 μL of dimethyl sulfoxide to each well, and measure the optical density of each well at a wavelength of 490 nm on a multifunctional microplate reader. Calculate the cell survival rate according to the following formula. When the cell survival rate is 50%, the corresponding drug concentration is the half inhibitory concentration IC 50 .

[0038]

[0039] The data were expressed as mean ± standard deviation (X ± S), and the data were analyzed using GraphPad Prism 5 statistical software. P < 0.05 was considered statistically significant. The inhibitory activity of the four compounds on human liver cancer cells HepG2 and SNU-739 was obtained as follows: Figure 2 shown.

[0040] Experimental Results

[0041] like Figure 2 As shown in the figure, the four compounds all had a certain inhibitory effect on HepG2 cells, and their cell half inhibitory concentration (IC 50 ) were 21.92±0.48μM, 26.23±1.06μM, 50.52±1.87μM, and 40.68±1.49μM, respectively. Compounds 1-4 also had a certain inhibitory effect on the growth of human liver cancer cell SNU-739 cells. Their IC 50They are: 28.19±0.13μM, 32.72±0.56μM, 51.85±2.33μM, and 96.37±1.27μM respectively.

[0042] The experimental results show that the nutmeg glycoside compounds 1, 2, 3, and 4 have good anti-tumor effects, especially anti-liver cancer effects, and can be used to prepare anti-tumor drugs, especially anti-liver cancer drugs.

Claims

1. Use of a nutmeg glycoside compound in the preparation of anti-tumor drugs.

2. The use according to claim 1, characterized in that The myristyl glucoside compound has at least one compound shown in the following structure; 。 3. The use according to claim 2, characterized in that The myristica fragrans glycoside compound is a mixture of compounds 1, 2, 3 and 4 in any proportion.

4. The use according to claim 1, characterized in that Application of nutmeg glycoside compounds in the preparation of anti-liver cancer drugs.

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