A Alpinetin Derivative, Its Preparation Method, Pharmaceutical Composition and Application

Abstract

The present invention discloses a galangin derivative, a preparation method thereof, a pharmaceutical composition and an application. The chemical name of the galangin derivative is (S)-7-propynyl-5-methoxydihydroflavone. The preparation method is as follows: Weigh galangin powder, add 3-bromopropyne, potassium carbonate and dichloromethane, stir at room temperature, add water after 12 h, extract with ethyl acetate, wash the obtained ethyl acetate extract successively with double distilled water and saturated brine, dry with anhydrous sodium sulfate, and then separate by silica gel column, and elute with petroleum ether-ethyl acetate solution gradient to obtain. The galangin derivative can be used in an anti-tumor pharmaceutical composition and has an anti-tumor effect. The galangin derivative of the present invention introduces an alkynyl group into the galangin structure, so that it achieves a better anti-tumor effect and has obvious anti-tumor activity against both liver cancer and lung cancer. Moreover, the preparation method is simple, the yield is high, the cost is low, and the preparation process is non-toxic and harmless, which is suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-tumor drugs, and particularly to an alpinetin derivative, a preparation method thereof, a pharmaceutical composition and an application thereof. Background Art

[0002] In recent years, due to the increase in the degree of aging worldwide and the existence of major risk factors such as local environmental pollution, the incidence of tumors has been increasing year by year, and the mortality rate of tumors has also been increasing year by year. Hepatocellular carcinoma (HCC) is the most common primary liver malignant tumor, accounting for 75% - 85% of primary liver cancers. Its incidence ranks fifth among global cancers. It is estimated that by 2025, more than 1 million people will be troubled by hepatocellular carcinoma every year. The high recurrence rate and strong ability of distant tissue metastasis of HCC lead to a short overall survival (OS) of patients and a poor prognosis, and it is the second leading cause of tumor-related death.

[0003] Currently, for the treatment of tumors, clinically, traditional means such as surgery, chemotherapy and radiotherapy, as well as immunotherapy are generally adopted. However, from the current cure rate, it is still difficult to achieve the goal of completely curing tumors. There is an urgent need to find more effective small molecules to better prevent and treat the occurrence and development of tumor diseases and reduce the incidence and mortality of tumor patients.

[0004] Alpinetin (ALP) is the main active monomer isolated from the seed mass of the natural Zingiberaceae plant Alpinia katsumadai. The chemical structure of alpinetin is 7-hydroxy-5-methoxy-flavanone, and the chemical structural formula is:

[0005]

[0006] The molecular weight (MW) is 270.28, and it belongs to flavonoid compounds. Because ALP can be isolated from a variety of Zingiberaceae plants and has the characteristics of low toxicity, good anti-tumor activity, wide therapeutic window, wide source distribution and low price, it has become a potential drug raw material for the research and development of anti-tumor diseases.

[0007] In order to develop chemical substances with better anti-tumor activity, the present application optimizes the structure of alpinetin, and obtains an alpinetin derivative with better anti-tumor activity through a reasonable and feasible preparation method, making the method reasonable and feasible, the route simple, the yield high, the cost low, without the use or generation of toxic and harmful substances, and enabling industrial production. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a galangin derivative, which can achieve better anti-tumor effects by introducing an alkynyl group into the structure of galangin. Moreover, the preparation method of this derivative is simple, with a high yield, low cost, non-toxic and harmless, and can realize industrial production.

[0009] The present invention provides a galangin derivative, and its chemical structural formula is:

[0010]

[0011] The chemical name of this galangin derivative is (S)-7-prop-2-ynyl-5-methoxydihydroflavone (5-methoxy-2-phenyl-7-(prop-2-yn-1-yloxy)chroman-4-one), and its molecular formula is C 19 H 16 O4, and the molecular weight (MW) is 308.3330.

[0012] As another improvement of the present invention, the present invention provides a preparation method of the galangin derivative. The preparation method is as follows: Weigh galangin powder, place it in a reaction flask, add 3-bromopropyne, potassium carbonate and dichloromethane, stir at room temperature, add water after 12 h, and extract with ethyl acetate. The obtained ethyl acetate extract is washed successively with double-distilled water and saturated brine, dried over anhydrous sodium sulfate, and then separated by a silica gel column, and eluted with a petroleum ether-ethyl acetate solution with a volume ratio of 100:1 - 1:1 in a gradient manner to obtain the galangin derivative.

[0013] Specifically, the amount of water added in the preparation method is the same as the amount of dichloromethane added, and the extraction times of ethyl acetate in the extraction step are at least 3 times.

[0014] The yield of the galangin derivative obtained by the method is greater than 80%.

[0015] As another improvement of the present invention, the present invention also provides an anti-tumor pharmaceutical composition. This anti-tumor pharmaceutical composition includes the above-mentioned galangin derivative or its pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier or excipient.

[0016] Among them, the pharmaceutically acceptable salts of alpinetin derivatives refer to the organic or inorganic addition salts of alpinetin derivatives, including but not limited to salts such as acetate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, edisylate, esylate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hippurate, hydrochloride, hydrobromide, hydroiodide, hydroxyethylsulfonate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthalenedicarboxylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / monohydrogenphosphate / dihydrogenphosphate, saccharate, stearate, succinate, tartrate, tosylate, trifluoroacetate, salts of aluminum, arginine, benzathine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, zinc, and their analogs.

[0017] The function of the "pharmaceutically acceptable carrier" is to transport the alpinetin derivative or its pharmaceutically acceptable salt so that it can play its due role. Therefore, the carrier must be compatible with the drug component, not affect the biological activity of the drug, and itself is relatively non-toxic and does not react with the drug it carries to cause toxic and side effects. Specifically, the carrier includes at least one of solvents, polymers, and liposomes. The solvents include but are not limited to water, physiological saline, and other non-aqueous solvents. The polymers include one or more of polylysine, polyethyleneimine and its modifications, polyamidoamine dendrimers and their derivatives, polypropyleneimine dendrimers and their derivatives, chitosan, polylactic acid, gelatin, cyclodextrin, and sodium alginate, but are not limited thereto. The liposome can be self-assembled from at least one of cationic lipids, cholesterol, and phospholipids. In the present invention, the alpinetin derivative or its pharmaceutically acceptable salt can be dispersed or adsorbed in the above carrier to form a dispersion system, or can be encapsulated by the above liposome or polymer to form a spherical structure. The alpinetin derivative or its pharmaceutically acceptable salt encapsulated in the spherical structure can be slowly released, controlled released, or targeted released to make it play the best efficacy, and can also improve the stability of the alpinetin derivative or its pharmaceutically acceptable salt and reduce drug irritation.

[0018] The function of "pharmaceutically acceptable excipients" is to fill the weight or volume of pharmaceutical dosage forms. Specifically, the excipients include at least one of diluents, excipients and stabilizers. The diluents include one or more of starches, sugars, celluloses and inorganic salts. The excipients refer to the additives other than the main anti-tumor drug active ingredients in anti-tumor drugs. The excipients include, for example, binders, fillers, disintegrants, lubricants in tablets, medicinal juices in pills, the matrix part in semi-solid preparations such as ointments and creams, and flavoring agents, cosolvents, emulsifiers, solubilizers, osmotic pressure regulators, coloring agents, etc. in liquid preparations. The main function of the stabilizers is to stabilize the active ingredients in anti-tumor drugs, and the stabilizers can be but are not limited to preservatives, antioxidants, cosolvents, emulsifiers, etc.

[0019] The pharmaceutical dosage forms of the anti-tumor drugs of the present invention include at least one of decoctions, powders, tablets, capsules, pills, oral preparations, granules, injections. Specific pharmaceutical dosage forms can be selected according to actual needs. The anti-tumor drugs are administered by oral or injection means. The injection is administered by intraperitoneal injection, subcutaneous injection, intramuscular injection or intravenous injection.

[0020] Of course, the pharmaceutical composition of the present invention can be made into conventional preparations, or can be made into sustained-release preparations, controlled-release preparations, targeted preparations and various particulate drug delivery systems.

[0021] As another improvement of the present invention, the present invention also provides the application of the above-mentioned alpinetin derivative in the preparation of anti-tumor drugs.

[0022] The tumors that the anti-tumor drugs can prevent or treat include but are not limited to liver cancer and lung cancer, and can also be solid tumors such as kidney cancer, pancreatic cancer, esophageal cancer, ovarian cancer, etc.

[0023] After adopting such a design, the present invention has at least the following advantages:

[0024] The alpinetin derivative of the present invention introduces an alkynyl group into the alpinetin structure, so that it achieves a better anti-tumor effect and has obvious anti-tumor activity against both liver cancer and lung cancer.

[0025] The preparation method of the alpinetin derivative of the present invention has simple steps, high yield, low cost, and is non-toxic and harmless during the preparation process, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The above is only an overview of the technical solution of the present invention. In order to be able to understand the technical means of the present invention more clearly, the following further describes the present invention in detail in conjunction with the drawings and specific embodiments.

[0027] Figure 1 is the synthetic reaction route diagram of the preparation method of the alpinetin derivative ALP-D of the present invention.

[0028] Figure 2 It is the positive ion mode mass spectrometry peak map of alpinetin derivative ALP-D of the present invention.

[0029] Figure 3 It is the analysis diagram of the nuclear magnetic resonance hydrogen spectrum results of alpinetin derivative ALP-D of the present invention.

[0030] Figure 4 It is the nuclear magnetic resonance carbon spectrum analysis diagram of alpinetin derivative ALP-D of the present invention.

[0031] Figure 5 It is the schematic diagram of the survival rate results of alpinetin and alpinetin derivative ALP-D on liver cancer cells in Example 2 of the present invention.

[0032] Figure 6 It is the schematic diagram of the survival rate results of alpinetin derivative ALP-D on tumor cells from different species sources in Example 2 of the present invention.

[0033] Figure 7 It is the apoptosis detection result diagram of alpinetin derivative ALP-D with different concentrations on tumor cells from different species sources in Example 3 of the present invention.

[0034] Figure 8 It is the result diagram of the inhibitory migration ability of alpinetin derivative ALP-D with different concentrations on different tumor cells in the cell migration experiment in Example 4 of the present invention.

[0035] Figure 9 It is the result diagram of the inhibitory effect of alpinetin and alpinetin derivative ALP-D on the formation of clone spheres of different tumor cells in the cell clone formation experiment in Example 4 of the present invention.

[0036] Figure 10 It is the rhodamine fluorescence imaging result diagram for the enrichment and labeling of target proteins by alpinetin derivative ALP-D in Example 5 of the present invention. Among them, the left side is the fluorescent gel, and the right side is the Coomassie brilliant blue staining.

[0037] Figure 11 It is the result of the protein mass spectrometry database search for the target proteins enriched by alpinetin derivative ALP-D in Example 5 of the present invention. Among them, the left side is a volcano plot showing proteins with a fold of change (FC) greater than two and a statistical p-value less than 0.05; the right side is six representative target proteins enriched.

[0038] Figure 12 It is the physical object comparison diagram of the inhibitory growth of tumor masses by alpinetin derivative ALP-D in Example 6 of the present invention. Detailed implementation manners

[0039] Example 1

[0040] 1. Preparation of Alpinetin Derivative ALP-D

[0041] Weigh 50 mg of alpinetin powder (0.18 mmol, purity > 99%), place it in a 100 mL reaction flask, add 220 mg of 3-bromopropyne, 256 mg of potassium carbonate, and 20 mL of dichloromethane. Stir at room temperature and use thin-layer chromatography to detect the reaction every 2 h until the reaction is complete. After 12 h, add 20 mL of water, extract with 20 mL of ethyl acetate 4 times, combine the ethyl acetate phases, wash successively with double-distilled water and saturated brine, and dry with anhydrous sodium sulfate. Then use silica gel column chromatography and elute with a gradient of petroleum ether - ethyl acetate (PE:EA = 100:1 - 1:1, v / v) to obtain 44 mg of white compound ALP-D with a yield of 88%.

[0042] Figure 1 The synthetic reaction route diagram of the alpinetin derivative ALP-D is shown. In this preparation method, the raw materials are simple and easily available, the steps are simple, no toxic or harmful substances need to be used, and no toxic or harmful substances will be generated, which is suitable for industrial production.

[0043] 2. Structural Analysis of Alpinetin Derivative ALP-D

[0044] Use high-resolution mass spectrometry (MS) to confirm the molecular weight of the product ALP-D, 1 1H-NMR and 13 13C-NMR to determine the structure of the derivative.

[0045] Figure 2 The positive-ion mode mass spectrometry peak diagram of the reaction product ALP-D is shown. The specific analysis results are as follows: HR ESI-MS m / z: 309.1111 [M + H]+ (molecular weight 309.73410, molecular formula C19H17O4); 331.0927 [M + Na]+ (molecular weight 331.3228, molecular formula C19H16NaO4); 639.1968 [2M + Na]+ (molecular weight 639.6558, molecular formula C38H32NaO8). That is, it shows that the molecular weight of the reaction product alpinetin derivative ALP-D is 308.3330, and the molecular formula is C 19 19 16 H17

[0046] Figure 3The analysis of the 1H NMR spectrum of the reaction product ALP-D is shown. The specific analysis results are as follows: 1H NMR (500 MHz, CDCl3) δH 7.36 - 7.46 (5H, overlap, H-2’, 3’, 4’, 5’, 6’), 6.23 (1H, d, J = 2.90 Hz, H-6), 6.16 (1H, d, J = 2.90 Hz, H-8), 5.40 (1H, dd, J = 16.45, 3.70 Hz, H-2), 4.69 (2H, d, J = 3.05 Hz, H-12), 3.89 (3H, s, H-11), 3.02 (1H, dd, J = 20.65, 16.45 Hz, H-3a), 2.98 (1H, s, H-14), 2.79 (1H, dd, J = 3.7, 20.65 Hz, H-3b).

[0047] Figure 4 The 13C NMR analysis of the reaction product ALP-D is shown. The specific analysis results are as follows: 13C NMR (125 MHz, CDCl3) δC 189.1 (C-4), 164.8 (C-7), 163.7 (C-9), 162.26 (C-5), 138.6 (C-1’), 128.7 (C-3’, 4’, 5’), 126.1 (C-2’, 6’), 106.4 (C-10), 94.4 (C-8), 93.6 (C-6), 79.2 (C-2), 77.4 (C-13), 76.4 (C-14), 56.2 (C-12), 55.9 (C-11), 45.5 (C-3).

[0048] Example 2 In vitro Tumor Cell Proliferation Toxicity Experiment

[0049] Mouse hepatoma cell line Hepa1-6, human hepatoma cell line HepG2, and human large cell lung cancer cell line H460 were respectively inoculated into 96-well plates, with about 5000 tumor cells in each well. After inoculation, they were placed in a cell culture incubator at 37 °C with 5% carbon dioxide for stable culture. Complete media containing gradient concentrations of 0, 25, 50, 100, 200, and 400 μM of crude alpinetin and the alpinetin derivative ALP-D solution were respectively prepared, and the above-mentioned tumor cells were treated with the prepared solutions for 48 h, and then replaced with DMEM medium containing 10% cell proliferation detection solution (cell counting kit-8, CCK-8), and incubated for another 3 h. The optical density (OD) value was measured at 450 nm using a microplate reader SpectraMax, and the cell growth curve was calculated and plotted according to the formula: cell survival rate = [(OD value of the experimental group - OD value of the blank group) / (OD value of the control group - OD value of the blank group)] × 100%.

[0050] The results are shown in the appendix Figure 5 and 6 . Figure 5 It shows the effect of the survival of human hepatoma cells HepG2 stimulated by galangin and galangin derivative ALP-D detected by the CCK-8 method Figure 6 It shows the effect of the survival of galangin derivative ALP-D on mouse hepatoma cells Hepa1-6 and human lung cancer cells H460

[0051] From Figure 5 It can be seen that after treating human hepatoma cells HepG2 with different concentrations of galangin and galangin derivative ALP-D for 48 h respectively, the IC50 value decreased significantly. It decreased from the half inhibitory concentration (IC50) value of 221 μM at 48 h to 61 μM. The results show that the activity of galangin derivative ALP-D is 3.6 times higher than that of galangin, and it has better killing activity than the original galangin drug

[0052] From Figure 6 It can be seen that after treating human lung cancer cells H460 and mouse hepatoma cells Hepa1-6 with different concentrations of galangin derivative ALP-D respectively, it showed an obvious concentration dependence, that is, as the concentration of galangin derivative ALP-D increased, the tumor cell survival rate decreased. The results show that galangin derivative ALP-D can have killing activity on tumor cells of different species sources

[0053] Example 3 Cell apoptosis experiment

[0054] Use the Annexin V-FITC / 7-AAD-PerCP cell apoptosis flow detection kit (Beyotime, Shanghai, China) to detect the cell apoptosis rate

[0055] Seed the human lung cancer cell line H460 and the mouse hepatoma cell line Hepa1-6 into 6-well plates, about 1×10 5 tumor cells per well, and treat them with different concentrations of galangin derivative ALP-D (concentrations are 10 μM and 50 μM respectively) and an equal amount of solvent control. After 24 h, wash three times with phosphate buffer solution (PBS, Invitrogen) without Ca 2+ / Mg 2+ ; then digest with 0.05% trypsin (Invitrogen) without EDTA for 5 min, and collect them into 1 mL of complete medium respectively to terminate the digestion. Leave the cell pellet by centrifugation at 1000 g for 5 min; then add PBS (without Ca 2+ / Mg 2+)After washing the cell pellet again, resuspend it in 195 μL of binding buffer; add 5 μL of Annexin V-FITC reagent to the cell suspension, gently vortex and incubate in the dark at room temperature for 30 min; add 5 μL of 7-AAD dye, mix gently, and immediately analyze apoptosis using a flow cytometer (BD Biosciences, CA, USA).

[0056] Figure 7 shows the apoptosis detection results of human lung cancer cells H460 and mouse liver cancer cells Hepa1-6 tumor cells by different concentrations of alpinetin derivative ALP-D. From Figure 7 it can be seen that after treatment with 10 μM and 50 μM of alpinetin derivative ALP-D respectively, compared with the solvent control group, the early apoptosis rate of H460 cells increased from 7.68% to 23.8% and 33.3% respectively, and the late apoptosis rate increased from 2.14% to 3.48% and 7.27% respectively; the early apoptosis rate of Hepa1-6 cells increased from 6.28% to 13.9% and 25.2% respectively, and the late apoptosis rate increased from 0.45% to 0.72% and 2.12% respectively. The results indicate that alpinetin derivative ALP-D can significantly induce apoptosis of tumor cells from different species sources and shows a concentration-dependent manner.

[0057] Experimental Example 4 Cell Migration and Colony Formation Assays

[0058] For the cell migration assay, calculate according to 10 4 Hepa1-6 tumor cells and H460 tumor cells per well, resuspend with 300 μL of serum-free cell medium, and inoculate into the upper chamber of the cell migration well. At the same time, add 500 μL of complete medium containing 10% fetal bovine serum (FBS) to the lower chamber; treat with different concentrations of alpinetin derivative ALP-D (10 and 50 μM) and an equal amount of solvent control respectively. After 24 h, perform cell migration measurement. Gently wipe off the non-migrated cells with a cotton swab, wash three times with PBS, fix the cells with cold methanol, add 0.4% crystal violet solution and stain for 30 min, and take pictures after washing clean with double-distilled water. The results are shown in the appendix Figure 8 .

[0059] For the cell colony formation assay, count 300 Hepa1-6 tumor cells, H460 tumor cells and HepG2 tumor cells per well, and inoculate them into 6-well plates or 12-well plates respectively. Treat with alpinetin and alpinetin derivative ALP-D (concentration of 50 μM) and an equal amount of solvent control for 14 days respectively. Wash three times with PBS, fix the cells with cold methanol, add 0.4% crystal violet solution and stain for 30 min, and take pictures after washing clean with double-distilled water. The results are shown in the appendix Figure 9 .

[0060] From Figure 8 It can be seen that alpinetin derivatives ALP-D with different concentrations can significantly inhibit the migration ability of H460 tumor cells and Hepa1-6 tumor cells.

[0061] From Figure 9 It can be seen that, compared with the solvent control group, after stimulation with a certain concentration of alpinetin and alpinetin derivative ALP-D, the formation of clone spheres of three kinds of tumor cells (HepG2, H460 and Hepa1-6) can be inhibited, and the inhibitory effect of alpinetin derivative ALP-D is particularly significant.

[0062] The results show that alpinetin derivative ALP-D can significantly inhibit the formation of clone spheres of tumor cells from different species sources, has a preventive effect on the occurrence of tumors from different species sources, and its effect is significantly better than that of the original alpinetin drug.

[0063] Experimental Example 5 Identification of the target bound by alpinetin derivative ALP-D

[0064] In this example, alpinetin derivative ALP-D can further introduce a reporter group through a bioorthogonal reaction (click chemistry reaction) by introducing a terminal alkyne group into the structure. There are two common types of reporter groups: the first is small molecule fluorescent groups, such as Rhodamine (Rho) and Cy3 / Cy5, etc. Such groups can be separated by SDS-PAGE and the proteins bound to the small molecules can be shown by a fluorescence imager; the second is Biotin group. After binding biotin to magnetic beads containing avidin, the enriched proteins are enzymatically digested into polypeptides, and then the peptide molecules are identified by high performance liquid chromatography-mass spectrometry (LC-MS / MS). The target protein bound by the drug can be determined based on the characteristic peptides.

[0065] In this example, after treating human hepatoma cells HepG2 with alpinetin derivative ALP-D and an equal amount of solvent control for 4 h, total proteins were extracted. Click chemical reactions were carried out with protein samples of equal concentration. Azido-rhodamine (20 μM), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine [TBTA, 0.1 mM], tris(2-carboxyethyl)phosphine hydrochloride [TCEP, 1 mM], and copper sulfate (CuSO4, 1 mM) were added in sequence. After mixing, the mixture was incubated on a shaker at 37 °C for 3 h; precipitated with cold methanol, the precipitate was washed twice, and the protein was dissolved in 40 μL of 0.4% SDS-PBS. After protein denaturation, SDS-PAGE electrophoresis was performed, and the protein labeling was observed using a fluorescence imager. The PAGE gel was stained with Coomassie Brilliant Blue staining solution. The results are shown in Appendix Figure 10 .

[0066] Similarly, after incubating human hepatoma cells HepG2 with alpinetin derivative ALP-D and an equal amount of solvent control for 4 h, total proteins were extracted. Click chemical reactions were carried out with protein samples of equal concentration. Azido-biotin (10 μM), TBTA (0.1 mM), TCEP (1 mM), and CuSO4 (1 mM) were added in sequence. After mixing, the mixture was incubated on a shaker at 37 °C for 3 h, precipitated with cold methanol, and the precipitate was washed twice; the protein was dissolved in 1 mL of 0.2% SDS-PBS, 50 μL of streptavidin beads was added, and the mixture was rotated at room temperature for 2 h and then centrifuged; the beads were washed with 1% SDS (3 times), urea (6 M, 2 times), and PBS (3 times) in sequence, followed by DTT reduction, alkylation, and digestion (16 h with 25 ng / μL trypsin). After desalting and freeze-drying the polypeptide sample, it was subjected to MS detection. The Uniprot human protein database was used for result analysis, and the candidate target enrichment was verified by Western blotting for the identification results. The results are shown in Appendix Figure 11 .

[0067] Figure 10 The rhodamine fluorescence imaging results are shown. The left side is the fluorescence gel, and the right side is the Coomassie Brilliant Blue staining. The results show that the alpinetin derivative ALP-D can enrich and label target proteins.

[0068] Figure 11 The target proteins enriched by the alpinetin derivative ALP-D shown in the protein mass spectrometry database search results are shown. The left side is a volcano plot showing proteins with a fold of change (FC) greater than two and a statistical p-value less than 0.05; the right side is six representative target proteins enriched.

[0069] Experimental Example 6 Evaluation of Mouse Transplanted Tumor Model

[0070] Male C57BL / 6 mice weighing about 18 - 22 g were selected, randomly grouped, given sufficient water and food, and all operations were carried out in a SPF clean environment. The animal experiments were approved by the Ethics Committee of Beijing Shijitan Hospital, Capital Medical University (Ethics number: 2023 - KY - 0009 - 001).

[0071] Specifically: A cell suspension of mouse hepatoma cells Hepa1 - 6 (about 10 6 cells) was mixed with high - concentration Matrigel at a ratio of 1:1 and operated on ice water; the cells were subcutaneously inoculated under the axilla of the mice, and the wound was sealed with collodion, and the mice were continued to be fed normally; after the tumor grew out, the size of the tumor mass was measured with a vernier caliper, and the volume (Tumor Volume, TV) calculation formula was: TV=(a * b 2 ) / 2, where a is the length of the tumor mass and b is the width of the tumor mass. When the tumor mass grew to about 100 mm 3 , alpinetin derivative ALP - D solution at 20 mg / kg was intraperitoneally injected or an equal dose of solvent was used as the control group, and the drug was administered three times a week. After 4 weeks of drug administration, the body weights of each group of mice were weighed, the tumor mass volume was calculated, and after the drug administration was completed, the mice were anesthetized, sacrificed, and the materials were taken, and the tumor weights were observed and weighed. The results are shown in Table 1 and Appendix Figure 12 .

[0072] Table 1 Results of Evaluation of Mouse Transplanted Tumor Model

[0073]

[0074]

[0075] Figure 12 The results show the anti - tumor activity of alpinetin derivative ALP - D evaluated by the in - vivo transplanted tumor model. It can be clearly seen from Table 1 and Figure 12 that after the intervention of alpinetin derivative ALP - D, the growth of the tumor mass in mice can be significantly inhibited, and the tumor mass has an obvious capsule and is in a poorly differentiated state. This shows that alpinetin derivative ALP - D has obvious anti - tumor activity.

[0076] The above - mentioned are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Those skilled in the art make some simple modifications, equivalent changes or decorations using the disclosed technical content above, and all fall within the protection scope of the present invention.

Claims

1. A galangin derivative, characterized in that, The chemical structure of the alpinetin derivative is as follows:

2. The preparation method of the alpinetin derivative according to claim 1, wherein, The method is as follows: Weigh alpinetin powder, place it in a reaction flask, add 3-bromopropyne, potassium carbonate and dichloromethane, stir at room temperature, add water after 12 h, extract with ethyl acetate, wash the obtained ethyl acetate extract successively with double-distilled water and saturated brine, dry over anhydrous sodium sulfate, and then separate by silica gel column, elute with a petroleum ether-ethyl acetate solution with a volume ratio of 100:1 - 1:1 in a gradient manner to obtain the alpinetin derivative.

3. The preparation method of the alpinetin derivative according to claim 2, characterized in that, In the method, the amount of water added is the same as the amount of dichloromethane added, and the number of extractions with ethyl acetate in the extraction step is at least 3 times.

4. The preparation method of the alpinetin derivative according to claim 3, characterized in that, The yield of the alpinetin derivative obtained by the method is greater than 80%.

5. An antitumor pharmaceutical composition, characterized in that, It includes the alpinetin derivative described in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

6. Use of the alpinetin derivative according to claim 1 in the preparation of an anti-tumor drug.

7. Use of the alpinetin derivative according to claim 6 in the preparation of an anti-tumor drug, characterized in that, The tumors that the anti-tumor drug can prevent or treat include but are not limited to liver cancer and lung cancer.

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