Iridaldehyde-type triterpenoid compound with anti-tumor effect, preparation method, composition and use thereof

Abstract

The present invention discloses an irisaldehyde-type triterpenoid compound with anti-tumor effect, a preparation method, a composition and a use thereof, and belongs to the technical field of chemical medicine. The compound is 16-methoxyiridogermenal, and its molecular formula is C 31 H 52 O4 was obtained by separating the ethyl acetate layer of the alcohol extract of Sichuan Belamcanda chinensis. The half-maximal inhibitory concentration of the compound 16-methoxyirisene on lung cancer cells, nasopharyngeal carcinoma cells and breast cancer cells was 39.44±3.48μmol·L ‑1 , 16.26±1.06μmol·L ‑1 , 36.26±1.30μmol·L ‑1 , has a significant inhibitory effect and can be used to prepare corresponding drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical drugs, and more particularly to an irisaldehyde-type triterpenoid compound with anti-tumor effect, a preparation method thereof, a composition and use thereof. Background Art

[0002] Iris tectorum (Chuanxia scutellaria) is the dried rhizome of Iris tectorum (Iris tectorum Maxim), a plant of the genus Iris in the family Iridaceae. Also known as blue butterfly, wild anemarrhena, and iron shoulder pole, it is primarily found in Guangdong, Guangxi, and Sichuan provinces. It was first mentioned in the Shennong Bencao Jing (Classic of Materia Medica) and is included in the 2005 edition of the Chinese Pharmacopoeia. Its medicinal properties are similar to those of Belamcanda dahurica, with a bitter and cold nature. It enters the lung meridian and has the effects of clearing heat and detoxifying, removing phlegm, and relieving sore throats. It is commonly used to treat symptoms such as heat-toxin stagnation, phlegm-fire accumulation, sore throat, excessive phlegm accumulation, cough, and shortness of breath. Clinically, it is primarily used to treat respiratory ailments such as upper respiratory tract infections, acute and chronic pharyngitis, bronchitis, and asthma.

[0003] For example, the Chinese patent with announcement number CN116098912A discloses a flavonoid glycoside combination for the treatment of chronic obstructive pulmonary disease, which combines irisin and trifolioside extracted from Belamcanda chinensis for the treatment of chronic obstructive pulmonary disease, achieving good results. However, the active substances in Belamcanda chinensis are complex and diverse and their effects are not clear. In order to further clarify the basis of the active substances in Belamcanda chinensis, the present invention studies the chemical components of Belamcanda chinensis, isolates and identifies a new chemical component of irisaldehyde-type triterpenoids from the alcohol extract of Belamcanda chinensis, and discovers its anti-cancer effect. After searching, the new chemical component of irisaldehyde-type triterpenoids of the present invention has not been disclosed by the prior art. For example, in the research progress of Belamcanda chinensis and the predictive analysis of its quality markers disclosed in Document 1, a triterpenoid compound extracted from Belamcanda chinensis is involved, but the structure is different from that of the present invention, and the anti-cancer effect is unclear.

[0004] Reference 1: Ling Yue et al. Research progress of Belamcanda chinensis and predictive analysis of its quality markers. Chinese Herbal Medicine, Vol. 53, No. 5, March 2022. Summary of the Invention

[0005] The first object of the present invention is to provide an irisaldehyde-type triterpenoid compound with anti-tumor effect, which is 16-methoxyiridogermenal, which has a significant inhibitory effect on lung cancer cells, nasopharyngeal carcinoma cells and breast cancer cells.

[0006] The second object of the present invention is to provide a method for preparing the irisaldehyde-type triterpenoid compound with anti-tumor effect, which is extracted from the alcohol extract using Sichuan Belamcanda chinensis as raw material.

[0007] The third object of the present invention is to provide an application of the irisaldehyde-type triterpenoid compound having anti-tumor effect in the preparation of lung cancer drugs.

[0008] The fourth object of the present invention is to provide a use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in the preparation of nasopharyngeal carcinoma drugs.

[0009] The fifth object of the present invention is to provide a use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in the preparation of breast cancer drugs.

[0010] The sixth object of the present invention is to provide an anti-tumor composition containing the irisaldehyde-type triterpenoid compound having anti-tumor effects.

[0011] In order to achieve these objectives of the present invention, the present invention provides an irisaldehyde-type triterpenoid compound with anti-tumor effect, which is 16-methoxyiridogermenal (16-methoxyisoiridogermenal), with a molecular formula of C 31 H 52 O4, the structure is as follows:

[0012]

[0013] The method for preparing the irisaldehyde-type triterpenoid compound with anti-tumor effect provided by the present invention comprises:

[0014] The medicinal material of Sichuan Belamcanda chinensis is extracted with ethanol and concentrated to obtain an ethanol extract concentrate;

[0015] The concentrated alcohol extract was extracted with petroleum ether, ethyl acetate and n-butanol in sequence to obtain extracts of each layer;

[0016] The ethyl acetate extract was subjected to silica gel column chromatography with a gradient elution of chloroform-methanol (50:1→30:1→20:1→10:1→8:1→5:1→1:1) to obtain 11 fractions, Fr.Y1 to Fr.Y11.

[0017] Fraction Fr.Y1 was further filtered, recrystallized, and chromatographed on a silica gel column using a gradient elution of petroleum ether-ethyl acetate (10:1→8:1→5:1→2:1→1:1). The fractions obtained by the 2:1 gradient elution were filtered, analyzed by TLC and HPLC, and combined into seven subfractions, Sub.Fr.Y1-1 to Sub.Fr.Y1-7.

[0018] The sub-fraction Sub.Fr.Y1-6 was separated by gel column, ODS column, silica gel column and preparative HPLC to obtain the irisaldehyde-type triterpenoid compound.

[0019] The present invention provides use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in preparing cancer drugs.

[0020] Specifically, the present invention provides the use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in the preparation of lung cancer drugs.

[0021] Specifically, the present invention provides the use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in the preparation of nasopharyngeal carcinoma drugs.

[0022] The present invention provides use of the irisaldehyde-type triterpenoid compound with anti-tumor effect in preparing breast cancer drugs.

[0023] The present invention provides an anti-tumor composition, which is composed of a prepared irisaldehyde-type triterpenoid compound and a phenylbenzofuran compound, wherein the mass ratio of the irisaldehyde-type triterpenoid compound to the phenylbenzofuran compound is 1:1; the phenylbenzofuran compound is a phenylbenzofuran compound disclosed in a previously applied Chinese patent (authorization publication number CN109516995B, publication date 2021-07-06), and its structure is:

[0024]

[0025] The present invention provides an application of the composition in preparing cancer drugs.

[0026] Specifically, the present invention provides a use of the composition in preparing lung cancer drugs.

[0027] Specifically, the present invention provides a use of the composition in preparing nasopharyngeal carcinoma drugs.

[0028] The present invention provides an application of the composition in preparing breast cancer drugs.

[0029] The present invention has at least the following beneficial effects:

[0030] 1. The present invention extracts a new irisaldehyde-type triterpenoid compound: 16-methoxyisoiridogermenal from Sichuan Belamcanda chinensis, which has a significant inhibitory effect on lung cancer cells, nasopharyngeal cancer cells and breast cancer cells.

[0031] 2. In one embodiment, the cell survival rate of 16-methoxyirisene of the present invention decreased significantly with increasing concentration, and the inhibitory effect was significantly enhanced on lung cancer A549 cells. Compared with the control group, the compound at 12.5 μmol·L -1 The inhibitory effect was significant at the concentration of 50%.50 ) values ​​were 39.44±3.48μmol·L -1 .

[0032] 3. In one embodiment, the cell survival rate of 16-methoxyirisene of the present invention decreased significantly with increasing concentration, and the inhibitory effect was significantly enhanced. Compared with the control group, the compound at 6.25 μmol·L -1 The half inhibitory concentration (IC 50 ) values ​​were 16.26±1.06μmol·L -1 .

[0033] 4. In one embodiment, the cell survival rate of 16-methoxyirisene of the present invention on breast cancer MDA-MB-231 cells decreased significantly with increasing concentration, and the inhibitory effect was significantly enhanced. Compared with the control group, the compound at 25 μmol·L -1 The inhibitory effect was significant at the concentration of 50%. 50 ) values ​​were 36.26±1.30μmol·L -1 .

[0034] 5. An anti-tumor composition of the present invention is composed of an irisaldehyde-type triterpenoid compound and a phenylbenzofuran compound. When the mass ratio of the irisaldehyde-type triterpenoid compound to the phenylbenzofuran compound is 1:1, it shows a significant inhibitory effect on lung cancer A549 cells, CNE-1 cells, and breast cancer MDA-MB-231 cells.

[0035] Other advantages, objectives and features of the present invention will be reflected in part from the following description and will be understood by those skilled in the art through study and practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 is the structural diagram of the compound 16-methoxyirisene of the present invention;

[0037] Figure 2 HMBC correlation diagram of the compound 16-methoxyirisene of the present invention;

[0038] Figure 3 is the NOESY correlation diagram of the compound 16-methoxyirisene of the present invention;

[0039] Figure 4is the survival rate (%) of lung cancer A549 cells after 48 hours of treatment with different concentrations of 16-methoxyirisene of the present invention;

[0040] Figure 5 is the survival rate (%) of nasopharyngeal carcinoma CNE-1 cells after 48 hours of treatment with different concentrations of 16-methoxyirisene of the present invention;

[0041] Figure 6 is the survival rate (%) of breast cancer MDA-MB-231 cells after 48 hours of treatment with different concentrations of 16-methoxyirisene of the present invention;

[0042] Figure 7 The survival rate (%) of nasopharyngeal carcinoma CNE-1 cells in drug group 1, drug group 2, and drug group 3 of the composition of the present invention at different concentrations after 48 hours;

[0043] Figure 8 The survival rate (%) of nasopharyngeal carcinoma CNE-1 cells in drug group 2 of different concentrations of the composition of the present invention after 48 hours;

[0044] Figure 9 The survival rate (%) of nasopharyngeal carcinoma CNE-1 cells in drug group 3 of different concentrations of the composition of the present invention after 48 hours;

[0045] Figure 10 The survival rate (%) of lung cancer A549 cells in drug group 2 of different concentrations of the composition of the present invention after 48 hours;

[0046] Figure 11 The figure is the survival rate (%) of breast cancer MDA-MB-231 cells in drug group 2 of different concentrations of the composition of the present invention after 48 hours. DETAILED DESCRIPTION

[0047] The present invention is further described in detail below with reference to examples so that those skilled in the art can implement the invention with reference to the description.

[0048] It should be understood that terms such as “having”, “including” and “comprising” used herein do not preclude the existence or addition of one or more other elements or combinations thereof.

[0049] It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0050] Example 1

[0051] 10 kg of dried Belamcanda chinensis was coarsely ground and extracted with 10-15 times the amount of 75% ethanol. The extraction was repeated twice, each time for 6-9 days. The filtrates from the two extractions were combined and the solvent was recovered under reduced pressure to obtain an alcohol extract concentrate. An appropriate amount of the concentrate was weighed to obtain approximately 3467 g of a total extract concentrate. The alcohol extract concentrate was extracted with petroleum ether, ethyl acetate, and n-butanol three times, using 10 L of each solvent each time. The extracted fractions were combined and concentrated to obtain a petroleum ether layer (CSS 6.8 g), an ethyl acetate layer (CSY 1557.08 g), an n-butanol layer (CSZ 403.2 g), and an aqueous fraction (CSW 1403.5 g).

[0052] Take 1500 g of ethyl acetate layer CSY, fully dissolve it, mix it with an equal amount of silica gel powder (80-100 mesh), and apply it to a silica gel column. Use chloroform-methanol system gradient elution with a gradient ratio of 50:1, 30:1, 20:1, 10:1, 8:1, 5:1, and 1:1. Finally, flush the column with methanol and water. Collect fractions according to column volume and combine them into 11 fractions, Fr.Y1 to Y11, according to TLC and HPLC analysis.

[0053] Among them, a large amount of light yellow solid was precipitated in fraction Fr.Y1. In order to ensure the smooth subsequent separation, the precipitated solid was filtered, dried and stored. Part of the solid was taken and repeatedly washed with alcohol, and recrystallized to obtain a white powder. It was filtered, and the filtrate (estimated weight 375.26 g) was gradient eluted with petroleum ether-ethyl acetate on a silica gel column with a gradient ratio of 10:1, 8:1, 5:1, 2:1, and 1:1. The distillate collected under the 2:1 gradient elution precipitated yellow crystals, which were filtered. The filtrate was analyzed by TLC and HPLC and combined into 7 sub-fractions Sub.Fr.Y1-1 to Sub.Fr.Y1-7.

[0054] Sub.Fr.Y1-6 (8.85 g) was purified by Sephadex LH-20 column, ODS column, silica gel column, and then by preparative HPLC (80% methanol water, 210 / 254 nm) to obtain a new compound (1330.1 mg, t R =20min).

[0055] Structural identification

[0056] The new compound obtained in Example 1: pale yellow oily liquid, with purple-red spots of sulfuric acid-vanillin. ESI-MS quasi-molecular ion peak m / z 511 [M+Na] + and 487[MH] - This indicates that its molecular weight is 488. 13 The C-NMR data and DEPT spectrum results showed that the compound had 31 carbon atom signals, consisting of 8×CH3, 10×CH2, 6×CH and 7×C. Its molecular formula was speculated to be C 31H 52 O4, has 6 degrees of unsaturation. The structural formula is Figure 1 shown.

[0057] Compound 1 H-NMR (CDCl3, 600MHz) spectrum (see Table 1) shows δ H 10.19 (1H, s) is the aldehyde carbonyl proton signal; three groups of olefin proton signals δ H 5.17 (1H, t, J = 7.4 Hz), δ H 5.08 (1H, t, J = 6.5 Hz) and δ H 5.01 (1H, t, J = 7.4 Hz); one sp 3 Proton signal δ on hybrid carbon H 3.61 (2H, t, J = 6.4 Hz); 7 methyl groups (δ H 1.84, 1.67, 1.59, 1.56, 1.45, 1.15, 1.10, each 3H, s). 13 C-NMR (CDCl3, 150MHz) and HSQC (see Table 2), δ C 189.9 (C-1) is an aldehyde carbonyl signal; δ C 162.6(C-7), δ C 136.4(C-19), δ C 134.6(C-15), δ C 133.2(C-2), δ C 131.2(C-23), δ C 128.0(C-14), δ C 124.3 (C-22), δ C 120.4 (C-18) is the signal of four groups of double bond carbons; δ C 75.0(C-10) and δ C 63.0 (C-3) is sp connected to oxygen 3 Hybridized carbon signal; δ C 26.3 (C-27), 25.7 (C-24), 17.9 (C-26), 17.7 (C-30), 16.2 (C-29), 10.9 (C-25), and 10.6 (C-28) are the seven methyl carbon signals.

[0058] According to the molecular formula, the unsaturation is calculated and it contains 6 unsaturations. Based on the above data, it is speculated that the compound has four groups of double bonds, one α,β-unsaturated aldehyde carbonyl group, and the remaining unsaturation, indicating that the compound may have a ring. Combined with the reference and NMR data comparison (Table 1 and Table 2), combined with HMBC analysis ( Figure 2 ), δ H 3.36(H-16) and δ C 55.8(-O C H3) is related to δ H 3.14(-OC H3 ) and δ C 87.3 (C-16) related, and δ H 5.17 (H-14) and δ H 5.01(H-18) and δ C 87.3 (C-16) has a long-range correlation, indicating that the -OCH3 is connected to C-16. In addition, in HMBC, the proton signal δ H 1.84 (3H, s, H-25) and double bond carbon δ C There is a long-range correlation between 189.9 (C-1) and 133.2 (C-2); the proton signal at the C-16 position δ H 3.36 (1H, t, J = 7.0 Hz, H-16) and δ H 1.45 (3H, s, H-28) and double bond tertiary carbon δ C 128.0 (C-14) related; proton signal δ H 1.56 (3H, s, H-29) and double bond carbon δ C 120.4 (C-18) and 136.4 (C-19) have long-range correlation; the proton signal δ H 1.67(3H,s,H-24),δ H 1.59(3H,s,H-30) and δ C There is a long-range correlation between 124.3 (C-22) and 131.2 (C-23). ​​The above correlation signals confirm the position of the α,β-unsaturated aldehyde carbonyl group and three groups of double bonds. Combined with the data in Tables 1 and 2, it indicates the structure of the double bond of the side chain at the C-11 position on the monoterpene ring of the compound.

[0059] According to NOESY ( Figure 3 ), the proton signal in the branched double bond δ H 1.56 (H-29) and proton signal δ H 2.10-2.28 (H-17), proton signal δ H 1.45 (H-28) and proton signal δ H 1.85-1.91 (H-13) have NOE correlations, suggesting that the double bond is Δ 18，19 , Δ 14,15 is in trans configuration. In addition, the proton signal δ in the six-membered ring H 2.01-2.05(H-5) and δ H2.55 (H-8β) correlation, δ H 1.85-1.86 (H-9α) and δ H The results showed that the H-6 structure of the ring was α-configuration and the H-26 structure was β-configuration. The methylated compound at the C-16 position had not been reported. The compound was determined to be a new compound and named 16-methoxyisoiridogermenal.

[0060] Table 1 Compounds 1 H-NMR (600 MHz) data (CDCl3, δ ppm)

[0061]

[0062] Table 2 Compounds 13 C-NMR (150 MHz) data (CDCl3, δ ppm)

[0063]

[0064]

[0065] Example 2

[0066] An antitumor composition comprises: the irisaldehyde-type triterpene compound (16-methoxyirisene) prepared in Example 1 and a phenylbenzofuran compound, wherein the mass ratio of the irisaldehyde-type triterpene compound to the phenylbenzofuran compound is 1:1.

[0067] The phenylbenzofuran compound is a phenylbenzofuran compound disclosed in a previously applied Chinese patent (authorization publication number CN109516995B, publication date 2021-07-06), and its structure is:

[0068] It has a certain inhibitory effect on nasopharyngeal carcinoma cells CNE-1 and CNE-2 and can be used for the treatment of nasopharyngeal carcinoma.

[0069] Example 3

[0070] 1. MTT assay to detect the cytotoxic activity of the irisaldehyde-type triterpenoid compound prepared in Example 1 and the anti-tumor composition of Example 2

[0071] The cytotoxic activity of irisaldehyde-type triterpenoid compound 16-methoxyirisene and the anti-tumor composition of Example 2 was tested by MTT assay. Lung cancer A549 cells, nasopharyngeal carcinoma CNE-1 cells, and breast cancer MDA-MB-231 cells were revived and passaged to the logarithmic growth phase. 4100 μL of each cell / well was inoculated into a 96-well plate, and the cell plate was cultured in a 37°C, 5% CO2 incubator. Drug intervention was performed after the cells attached (24 hours). A control group (no drug), a drug group, and a blank group without cells were set up. The drug addition concentrations in the groups were 6.25, 12.5, 25, 50, and 100 μmol·L -1 Continue incubation for 48 h. After 48 h, add 20 μL of MTT solution (5 mg mL -1 ), and then incubate in the dark for 4 hours. Dissolve the blue-purple crystals with 150 μL of DMSO, shake horizontally for 10 minutes, and measure the A (absorbance) value at a wavelength of 490 nm. According to the A (absorbance) value, the experimental data were statistically analyzed using Graphpad Prism 9.0 software (CA, USA), and the results were expressed as mean ± SD. Statistical analysis was performed by one-way variance to compare the differences between the groups, where p < 0.05 was considered to be statistically significant between the two groups. Calculate cell survival rate / half inhibitory concentration (IC 50 )value.

[0072]

[0073] The drug groups are:

[0074] 1. Irisaldehyde-type triterpenoid compound (16-methoxyirisene) drug group;

[0075] 2. Composition drug group 1 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 4:1);

[0076] 3. Composition drug group 2 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1);

[0077] 4. Composition drug group 3 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:4).

[0078] II. Evaluation of the cytotoxic activity of irisaldehyde-type triterpenoid 16-methoxyirisene

[0079] 1. The MTT method was used to determine the cytotoxic activity of the compound 16-methoxyirisene against lung cancer A549 cells. Figure 4 ) showed that as the concentration of 16-methoxyirisene increased, the cell survival rate decreased significantly and the inhibitory effect was significantly enhanced. Compared with the control group, the compound at 12.5 μmol·L -1 The inhibitory effect was significant at the concentration of 50%. 50) values ​​were 39.44±3.48μmol·L -1 In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0080] 2. The MTT method was used to determine the cytotoxic activity of the compound 16-methoxyirisene against nasopharyngeal carcinoma CNE-1 cells. Figure 5 ) showed that as the concentration of 16-methoxyirisene increased, the cell survival rate decreased significantly and the inhibitory effect was significantly enhanced. Compared with the control group, the compound at 6.25μmol·L -1 The half inhibitory concentration (IC 50 ) values ​​were 16.26±1.06μmol·L -1 In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0081] 3. The MTT assay was used to determine the cytotoxic activity of the compound 16-methoxyirisene against breast cancer MDA-MB-231 cells. Figure 6 ) showed that as the concentration of 16-methoxyirisene increased, the cell survival rate decreased significantly and the inhibitory effect was significantly enhanced on breast cancer MDA-MB-231 cells. Compared with the control group, the compound at 25 μmol·L -1 The inhibitory effect was significant at the concentration of 50%. 50 ) values ​​were 36.26±1.30μmol·L -1 In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0082] 4. The MTT method was used to determine the cytotoxic activity of the drug combination group 1, drug combination group 2, and drug combination group 3 on nasopharyngeal carcinoma CNE-1 cells. Figure 7 、 Figure 8 and Figure 9 As shown, in the three drug combination groups, the cell survival rate decreased significantly and the inhibitory effect increased significantly with the increase of drug concentration.

[0083] like Figure 7 As shown, compared with the control group, the drug group 1 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 4:1) had a -1The inhibitory effect was significant at the concentration of 50%. 50 ) values ​​were 21.97±1.16μmol·L -1 .

[0084] like Figure 8 As shown, compared with the control group, the drug group 2 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1) was -1 The inhibitory effect was significant at a concentration of 50 ) values ​​were 9.00±0.36μmol·L -1 .

[0085] like Figure 9 As shown, compared with the control group, the drug group 3 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:4) at 12.5 μmol·L -1 The inhibitory effect was significant at the concentration of 50%. 50 ) values ​​were 29.52±0.48μmol·L -1 .

[0086] In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0087] From the comparison of composition drug group 1 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 4:1), composition drug group 2 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1) and composition drug group 3 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:4), it can be seen that when the ratio of irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1, its inhibitory effect on nasopharyngeal carcinoma CNE-1 cells is significantly better than other ratios.

[0088] 5. The cytotoxic activity of drug group 2 against lung cancer A549 cells was determined by MTT assay. Figure 10 As shown in Figure 2, with the increase of drug concentration, the survival rate of lung cancer A549 cells decreased significantly, and the inhibitory effect was significantly enhanced. Combination drug group 2 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1) at 6.25 μmol·L -1 The half inhibitory concentration (IC 50) values ​​were 19.15±1.89μmol·L -1 In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0089] 6. The cytotoxic activity of drug group 2 against breast cancer MDA-MB-231 cells was determined using the MTT assay. Figure 11 As shown in the results, the drug group 2 (irisaldehyde-type triterpenoid compound: phenylbenzofuran compound = 1:1) showed a significant decrease in cell survival rate and a significant increase in inhibitory effect on breast cancer MDA-MB-231 cells with increasing concentration. Compared with the control group, the compound at 12.5 μmol·L -1 The inhibitory effect was significant at the concentration of 50%. 50 ) values ​​were 26.11±0.91μmol·L -1 In the figure, compared with the control group: *P<0.05; **P<0.01; ***P<0.001.

[0090] Although the embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiment. They can be applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily realized.

Claims

1. A method for preparing an irisaldehyde-type triterpenoid compound having anti-tumor effect, characterized in that: The structure of the compound is as follows: ； The preparation steps include: The medicinal material of Sichuan Belamcanda chinensis is extracted with ethanol and concentrated to obtain an ethanol extract concentrate; The concentrated alcohol extract was extracted with petroleum ether, ethyl acetate and n-butanol in sequence to obtain extracts of each layer; The ethyl acetate layer extract was subjected to silica gel column chromatography using a chloroform-methanol gradient elution with a gradient ratio of chloroform:methanol = 50:1, 30:1, 20:1, 10:1, 8:1, 5:1, and 1:1 to obtain 11 fractions Fr.Y1 to Fr.Y11; Fraction Fr.Y1 was filtered, recrystallized, and chromatographed again on a silica gel column using a petroleum ether-ethyl acetate gradient elution. The gradient ratios were 10:1, 8:1, 5:1, 2:1, and 1:

1. Fractions obtained from the 2:1 gradient elution were filtered, analyzed by TLC and HPLC, and combined into seven subfractions, Sub. Fr.Y1-1 to Sub. Fr.Y1-7. The sub-fraction Sub. Fr. Y1-6 was sequentially separated by gel column, ODS column, silica gel column and preparative HPLC to obtain the irisaldehyde-type triterpenoid compound.

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