Indole alkaloid derivatives from Brucea javanica and their preparation method and anti-tumor application

Abstract

The present invention relates to indole alkaloids from Brucea javanica, their preparation methods, and applications, belonging to the field of pharmaceutical technology. Specifically, the invention relates to two indole alkaloid derivatives extracted and isolated from Brucea javanica, a plant of the genus Simarthaceae. These derivatives exhibit excellent anti-tumor activity. The preparation method of the present invention is simple and easy to implement, has good reproducibility, and is of high purity. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and particularly relates to two indole alkaloid derivatives prepared from Brucea javanica fruit, a preparation method thereof, and applications thereof in anti-tumor aspects. Background Art

[0002] Brucea javanica (Brucea javanica) is the dried mature seed of Brucea javanica (Linnaeus) Meril, a plant of the Simaroubaceae family. Also known as Brucea javanica, Kushenzi, and Crow's Egg, Brucea javanica is primarily produced in Guangdong, Guangxi, Hainan, Yunnan, Fujian, and Taiwan in my country. Brucea javanica is bitter, cold, and slightly toxic. It enters the large intestine and liver meridians and has benefits such as clearing heat, detoxifying, stopping malaria, stopping dysentery, and corroding warts. Modern research indicates that Brucea javanica contains a variety of chemical components, including quasin, alkaloids, triterpenes, and steroids.

[0003] The novel indole alkaloid derivatives extracted and separated by the present invention and their anti-tumor activities have not been reported in any patent or non-patent literature so far. Summary of the Invention

[0004] The purpose of the present invention is to address the deficiencies of the prior art and to provide two indole alkaloid derivatives and methods for preparing the same. The present invention also provides the use of the indole alkaloid derivatives in anti-tumor applications.

[0005] In order to achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0006] In the first aspect, the two indole alkaloid derivatives of the present invention, isolated from Brucea javanica, have the following structures:

[0007]

[0008] In a second aspect, the preparation method of the present invention comprises the steps of:

[0009] The dried Brucea javanica seed powder defatted with petroleum ether was extracted with ethanol, and the combined extracts were concentrated to obtain an extract. The extract was extracted with n-butanol and the resulting fractions were chromatographed on a silica gel column using a dichloromethane-methanol system (1:0-0:1) for gradient elution. A total of 6 fractions, Fr.1-6, were collected.

[0010] Component Fr.2 was eluted using HP20, ODS column chromatography with an ethanol-water system (20:80-100:0) by gradient elution to obtain six components Fr.A-Fr.F;

[0011] Compound 1-2 was obtained by silica gel column chromatography eluting with a petroleum ether-ethyl acetate system of 100:1-1:1 and separation of Fr.E by preparative reverse-phase high performance liquid chromatography using an acetonitrile-water mobile phase.

[0012] The method for preparing the indole alkaloid derivatives in Brucea javanica is characterized in that the Brucea javanica used is Brucea javanica of the genus Simaroniae in the family Simaraceae.

[0013] The method for preparing the indole alkaloid derivatives in Brucea javanica is characterized in that the dried Brucea javanica seed powder defatted with petroleum ether is extracted with 80% industrial ethanol under reflux for three times, each time for 2-3 hours.

[0014] The method for preparing the indole alkaloid derivatives in Brucea javanica is characterized in that Fr.E is separated using an acetonitrile-water mobile phase with a ratio of 50:50 to 20:80.

[0015] The obtained compound was systematically identified and the results are as follows:

[0016] The structure of compound 1-2 was identified by high resolution mass spectrometry, one-dimensional NMR and two-dimensional NMR. The corresponding characterization data are shown in Table 1 and the spectrum is shown in Figure 1-10 shown.

[0017] Yadanziindole A (1): white crystals. (c0.1, methanol); HRESIMS: [M+H] + (m / z): 204.0913 The molecular formula was confirmed to be C 12 H 13 NO2, by analyzing yadanziindoleA 1 HNMR, 13 CNMR, HSQC and HMBC spectra confirmed the structure of yadanziindoleA, which is a new compound.

[0018] Yadanziindole B (2): white crystals. (c 0.1, methanol); HRESIMS: [M+Na] + (m / z): 242.0789 The molecular formula was confirmed to be C 12 H 13 NO3, by analyzing yadanziindoleB 1 HNMR, 13 CNMR, HSQC and HMBC spectra confirmed the structure of yadanziindole B, which is a new compound.

[0019] The anti-tumor activities of the two new compounds described in the present invention were investigated. Compounds 1 and 2 showed good anti-tumor activities. Therefore, the novel indole alkaloid derivatives described in the present invention have the prospect of being further developed into drugs for preventing and treating tumors.

[0020] In a third aspect, the present invention provides a pharmaceutical composition comprising the indole alkaloid derivative prepared from Brucea javanica or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0021] In a fourth aspect, the present invention further provides the use of the indole alkaloid derivative prepared from Brucea javanica or its pharmaceutically acceptable salt or a pharmaceutical composition comprising the derivative in the preparation of anti-tumor drugs.

[0022] Preferably, the tumor is a solid tumor.

[0023] More preferably, the tumor is lung cancer, breast cancer or liver cancer.

[0024] Compared with the prior art, the present invention has the following beneficial effects: The advantages of the present invention are that the compounds are all new compounds, which are indole alkaloid compounds with novel structures. At the same time, compounds 1 and 2 have good anti-tumor activity and are worthy of further development.

[0025] Table 1. Compound 1-2 1 H NMR and 13 C NMR data (600 MHz, in DMSO-d6, δ in ppm)

[0026] BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 HRESIMS spectrum of compound 1;

[0028] Figure 2 Compound 1 1 H-NMR spectrum;

[0029] Figure 3 Compound 1 13 C-NMR spectrum;

[0030] Figure 4 HSQC spectrum of compound 1;

[0031] Figure 5 HMBC spectrum of compound 1;

[0032] Figure 6 HRESIMS spectrum of compound 2;

[0033] Figure 7Compound 2 1 H-NMR spectrum;

[0034] Figure 8 Compound 2 13 C-NMR spectrum;

[0035] Figure 9 HSQC spectrum of compound 2;

[0036] Figure 10 HMBC spectrum of compound 2. DETAILED DESCRIPTION

[0037] The following examples are intended to help those skilled in the art better understand the present invention, but are not intended to limit the present invention in any way.

[0038] Example 1: Preparation of Compound 1-2

[0039] The dried Brucea javanica seed powder defatted with petroleum ether was extracted with ethanol, and the combined extracts were concentrated to obtain an extract. The extract was extracted with ethyl acetate and the resulting components were chromatographed on a silica gel column using a dichloromethane-methanol system (1:0-0:1) for gradient elution. A total of 6 components, Fr.1-6, were collected.

[0040] The pigment of component Fr.2 was removed by HP20 column chromatography using an ethanol-water system of 20:80-100:0.

[0041] Component Fr.2 was eluted using an ODS column chromatography gradient with an ethanol-water system of 20:80-100:0 to obtain six components Fr.A-Fr.F;

[0042] Compound 1-2 was obtained by silica gel column chromatography eluting with a petroleum ether-ethyl acetate system of 100:1-1:1 and preparative reverse-phase high performance liquid chromatography using a mobile phase of 50:50-20:80 acetonitrile-water to separate Fr.E.

[0043] Example 2: Investigation of the anti-tumor activity of compound 1-2

[0044] The growth inhibitory activity of compounds 1-2 on tumor cell lines A549, HepG2 and MCF-7 (ATCC, Rockville, MD, USA) was tested using the MTT assay. 4 Each well was inoculated with 100 μL of the test compound per well in a 96-well plate, and the wells were set up in triplicate and incubated for 24 hours at 37°C in a 5% CO2 incubator. The test compound was then treated with different concentrations (0.1, 1, 10, 25, and 50 μM) for 48 hours.

[0045] After 48 hours of drug exposure, cells were incubated with MTT solution (0.5 mg / mL) at 37°C for an additional 4 hours. After aspirating the culture medium, DMSO was added to each well to completely dissolve the cells, and absorbance was measured at 490 nm using a microplate reader. All experiments were performed in triplicate, with blank and positive control groups as shown in Table 2.

[0046] Table 2 Inhibitory effect of indole alkaloids in the present invention on the proliferation of human lung cancer, breast cancer and liver cancer cells

[0047]

[0048] The experimental results showed that compounds 1 and 2 exhibited good anti-tumor activity, and the novel indole alkaloid derivatives of the present invention have the prospect of being further developed into drugs for preventing and treating tumors.

[0049] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. An indole alkaloid derivative from Brucea javanica, characterized in that: The indole alkaloid derivative has any one of the following structures:

2. A method for preparing the indole alkaloid derivatives in Brucea javanica according to claim 1, characterized in that: The steps include: The dried Brucea javanica seed powder defatted with petroleum ether was taken, the extracts were combined and concentrated to obtain an extract, the extract was extracted with ethyl acetate, and the obtained components were subjected to silica gel column chromatography with a dichloromethane-methanol system of 1:0-0:1 gradient elution to obtain 6 components Fr.1-6; Component Fr.2 was eluted using HP20, ODS column chromatography with an ethanol-water system (20:80-100:0) by gradient elution to obtain six components, Fr.A-Fr.F. Compound 1-2 was obtained by silica gel column chromatography eluting with a petroleum ether-ethyl acetate system of 100:1-1:1 and separation of Fr.E by preparative reverse-phase high performance liquid chromatography using an acetonitrile-water mobile phase.

3. The method for preparing the indole alkaloid derivatives from Brucea javanica according to claim 2, characterized in that: The Brucea javanica used is Brucea javanica of the genus Simaroniae in the family Simarthaceae.

4. The method for preparing the indole alkaloid derivatives from Brucea javanica according to claim 2, characterized in that: The dried Brucea javanica seed powder defatted with petroleum ether was extracted with 80% industrial ethanol under reflux for 3 times, each time for 2-3 hours.

5. The method for preparing the indole alkaloid derivatives from Brucea javanica according to claim 2, characterized in that: Fr.E was separated using 50:50-20:80 acetonitrile-water mobile phase.

6. A pharmaceutical composition, characterized in that The invention comprises the indole alkaloid derivative in the Brucea javanica fruit as claimed in claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

7. Use of the indole alkaloid derivatives of Brucea javanica or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of drugs for treating lung cancer or liver cancer.

8. Use of the pharmaceutical composition according to claim 6 in the preparation of drugs for treating lung cancer or liver cancer.

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