Alpinetin derivative as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention discloses an alpinetin derivative as well as a preparation method, a pharmaceutical composition and application thereof. The chemical name of the alpinetin derivative is (S)-7-propargyl-5-methoxy flavanone, and the chemical name of the alpinetin derivative is (S)-7-propargyl-5-methoxy flavanone. The preparation method comprises the following steps: weighing alpinia powder, adding 3-propargyl bromide, potassium carbonate and dichloromethane, stirring at room temperature, adding water after 12 hours, extracting with ethyl acetate, cleaning the obtained ethyl acetate extract with double distilled water and saturated saline solution in sequence, drying with anhydrous sodium sulfate, separating with a silica gel column, and carrying out gradient elution with petroleum ether-ethyl acetate liquid to obtain the alpinia extract. The alpinetin derivative can be used for an anti-tumor pharmaceutical composition and has an anti-tumor effect. According to the alpinetin derivative disclosed by the invention, an alkynyl group is introduced into an alpinetin structure, so that the alpinetin derivative achieves a better anti-tumor effect and has obvious anti-tumor activity on both liver cancer and lung cancer. And the preparation method is simple, high in yield, low in cost, non-toxic and harmless in the preparation process and suitable for industrial production.

Description

Technical Field

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

[0002] In recent years, due to the increasing aging of the world and the existence of major risk factors such as local environmental pollution, the incidence of tumors has increased year by year, and the mortality rate of tumors has also increased year by year. Hepatocellular carcinoma (HCC) is the most common primary liver malignancy, accounting for 75% to 85% of primary liver cancer, and its incidence rate ranks fifth among cancers in the world. It is estimated that by 2025, more than 1 million people will be troubled by hepatocellular carcinoma each year. HCC has a high recurrence rate and strong ability to metastasize to distant tissues, resulting in a short overall survival (OS) and poor prognosis of patients, making it the second leading cause of tumor-related death.

[0003] Currently, the treatment of tumors in clinical practice generally adopts traditional methods such as surgery, chemotherapy and radiotherapy, as well as immunotherapy. However, judging 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 separated from the seeds of the natural ginger plant Alpinia officinalis. The chemical structure of Alpinetin is 7-hydroxy-5-methoxy-flavanone, and the chemical formula is:

[0005]

[0006] The molecular weight (MW) is 270.28, belonging to the flavonoids. ALP can be isolated from a variety of ginger plants and has the characteristics of low toxicity, good anti-tumor activity, wide therapeutic window, wide distribution of sources and low price, making it a potential drug for the development of anti-tumor drugs.

[0007] In order to develop chemical substances with better anti-tumor activity, the present application optimizes the structure of shanpin and obtains shanpin derivatives with better anti-tumor activity through a reasonable and feasible preparation method, so that the method is reasonable and feasible, the route is simple, the yield is high, the cost is low, no toxic and harmful substances are used or produced, and industrial production can be realized. Summary of the invention

[0008] The technical problem to be solved by the present invention is to provide a shanjiange derivative which can achieve a better anti-tumor effect by introducing an alkynyl group into the shanjiange structure, and the preparation method of the derivative is simple, high in yield, low in cost, and the preparation method is non-toxic and harmless, and can be industrialized.

[0009] The present invention provides a shanpin derivative, the chemical structural formula of which is:

[0010]

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

[0012] As another improvement of the present invention, the present invention provides a preparation method of the sanpin derivative, which comprises the following steps: weighing sanpin powder, placing it in a reaction bottle, adding 3-bromopropyne, potassium carbonate and dichloromethane, stirring at room temperature, adding water after 12 hours, and extracting with ethyl acetate, washing the obtained ethyl acetate extract with double distilled water and saturated brine in turn, drying with anhydrous sodium sulfate, separating with a silica gel column, and eluting with a gradient of petroleum ether-ethyl acetate with a volume ratio of 100:1-1:1 to obtain the sanpin derivative.

[0013] Specifically, in the preparation 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.

[0014] The yield of the shanpin 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, which comprises the above-mentioned sanpin derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0016] The pharmaceutically acceptable salt of sanpin derivatives refers to organic or inorganic addition salts of sanpin derivatives. Examples include, but are not limited to, acetate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, edisylate, ethoxylate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hexabenzoate, hydrochloride, hydrobromide, hydroiodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthalene, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, sucrose, stearate, succinate, tartrate, toluenesulfonate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, zinc, and the like.

[0017] The role of the "pharmaceutically acceptable carrier" is to transport the shanjiangsu derivative or its pharmaceutically acceptable salt so that it can play its due role. Therefore, the carrier must be compatible with the drug ingredients, 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 side effects. Specifically, the carrier includes at least one of a solvent, a polymer and a liposome. The solvent includes but is not limited to water, physiological saline, and other non-aqueous solvents. The polymer includes one or more of polylysine, polyethyleneimine and its modified products, polyamide-amine dendrimers and their derivatives, polypropyleneimine dendrimers and their derivatives, chitosan, polylactic acid, gelatin, cyclodextrin and sodium alginate, but is not limited to this. The liposome can be self-assembled from at least one of cationic lipids, cholesterol and phospholipids. In the present invention, the shanjiangsu derivative or its pharmaceutically acceptable salt can be dispersed or adsorbed in the above-mentioned carrier to form a dispersed system, or it can be encapsulated by the above-mentioned liposomes and polymers to form a spherical structure. The sanpin derivative or its pharmaceutically acceptable salt encapsulated in the spherical structure can be sustained-released, controlled-released or targeted-released to achieve optimal efficacy, improve the stability of the sanpin 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 the drug dosage form. 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 additives in anti-tumor drugs other than the main active ingredients of anti-tumor drugs. The excipients include adhesives, fillers, disintegrants, lubricants in tablets, juices in pills, etc., matrix parts in semi-solid preparations ointments and creams, and fragrances, solubilizers, emulsifiers, solubilizers, osmotic pressure regulators, colorants, etc. in liquid preparations. The main function of the stabilizer is to stabilize the active ingredients in the anti-tumor drug. The stabilizer can be, but is not limited to, preservatives, antioxidants, solubilizers, emulsifiers, etc.

[0019] The pharmaceutical dosage form of the anti-tumor drug of the present invention includes at least one of decoction, powder, tablet, capsule, pill, oral agent, granule, and injection. The specific pharmaceutical dosage form can be selected according to actual needs. The anti-tumor drug is administered orally or by injection. 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 a common preparation, or can be made into a sustained-release preparation, a controlled-release preparation, a targeted preparation and various microparticle drug delivery systems.

[0021] As another improvement of the present invention, the present invention also provides the use of the above-mentioned shanpin derivatives in the preparation of anti-tumor drugs.

[0022] The tumors that can be prevented or treated by the anti-tumor drug include, but are not limited to, liver cancer and lung cancer, and may also be solid tumors such as kidney cancer, pancreatic cancer, esophageal cancer, and ovarian cancer.

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

[0024] The sanpin derivative of the present invention achieves a better anti-tumor effect by introducing an alkynyl group into the sanpin structure, and has obvious anti-tumor activity against liver cancer and lung cancer.

[0025] The preparation method of the shanpin derivative of the invention has simple steps, high yield, low cost, non-toxic and harmless 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 more clearly understand the technical means of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0027] Figure 1 The invention discloses a synthetic reaction route for the preparation of the alpinia officinalis derivative ALP-D.

[0028] Figure 2 It is the positive ion mode mass spectrum peak diagram of the shanjiangsu derivative ALP-D of the present invention.

[0029] Figure 3 It is an analysis chart of the nuclear magnetic hydrogen spectrum results of the shanjiangsu derivative ALP-D of the present invention.

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

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

[0032] Figure 6 This is a schematic diagram of the results of the effect of the alpinia officinalis derivative ALP-D on the survival rate of tumor cells from different species in Example 2 of the present invention.

[0033] Figure 7 This is a graph showing the apoptosis detection results of different species-derived tumor cells subjected to different concentrations of the alpinia officinalis derivative ALP-D in Example 3 of the present invention.

[0034] Figure 8 This is a graph showing the ability of different concentrations of the alpinia officinalis derivative ALP-D to inhibit the migration of different tumor cells in a cell migration experiment in Example 4 of the present invention.

[0035] Fig. 9 This is a graph showing the results of the inhibition of clonal sphere formation of different types of tumor cells by shanjiangin and shanjiangin derivative ALP-D in a cell cloning experiment in Example 4 of the present invention.

[0036] Fig.10 This is a rhodamine fluorescence imaging result diagram of the enrichment and labeling of the target protein by the shanjiangsu derivative ALP-D in Example 5 of the present invention, wherein the left side is fluorescent gel and the right side is Coomassie brilliant blue staining.

[0037] Fig.11 This is the result of the protein spectrum search of the target protein enriched by the shanjiangsu derivative ALP-D in Example 5 of the present invention. The left side is a volcano plot showing proteins with a differential expression fold change (FC) greater than two times and a statistical p value less than 0.05; the right side shows six representative target proteins enriched.

[0038] Fig.12 This is a comparison of the actual tumor masses of mice in which the growth inhibition of the alpinia officinalis derivative ALP-D in Example 6 of the present invention is shown. DETAILED DESCRIPTION

[0039] Example 1

[0040] 1. Preparation of Alpinia officinalis derivative ALP-D

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

[0042] Figure 1 The synthetic reaction route of the shanpin derivative ALP-D is shown. The raw materials in the preparation method are simple and easy to obtain, the steps are simple, no toxic and harmful substances are used, and no toxic and harmful substances are produced, so the method is suitable for industrial production.

[0043] 2. Structural analysis of the galangin derivative ALP-D

[0044] The molecular weight of the product ALP-D was confirmed by high-resolution mass spectrometry (MS). 1 H-NMR and 13 The structure of the derivative was confirmed by C-NMR.

[0045] Figure 2 The positive ion mode mass spectrum peak of the reaction product ALP-D is shown, and the specific analysis results are: 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, the shanjiangsu derivative ALP-D, is 308.3330, and the molecular formula is C 19 H 16 O4.

[0046] Figure 3The results of H-NMR spectrum analysis of the reaction product ALP-D are shown. The specific analysis results are: 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 NMR carbon spectrum analysis of the reaction product ALP-D is shown. The specific analysis results are: 13CNMR (125MHz, CDCl3) δC 189.1 (C-4), 164.8 (C-7), 163.7 (C-9), 162.2.6 (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 liver cancer cell line Hepa1-6, human liver cancer cell line HepG2 and human large cell lung cancer cell line H460 were inoculated into 96-well plates, with about 5000 tumor cells per well, and then placed in a cell culture incubator at 37°C with 5% carbon dioxide for stable culture. Complete culture medium containing gradient concentrations of 0, 25, 50, 100, 200 and 400 μM of shanjiangsu original drug and shanjiangsu derivative ALP-D solution was prepared respectively, and the above tumor cells were treated with the prepared solution for 48 hours, and then replaced with DMEM medium containing 10% cell proliferation detection solution (cell counting kit-8, CCK-8), and continued to incubate for 3 hours. The optical density (OD) value was measured at 450nm using a microplate reader SpectraMax, and the cell growth curve was calculated and drawn according to the formula of 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] See attached for the results Figure 5 and 6 . Figure 5 The figure shows the use of CCK-8 method to detect the effects of ALP-D stimulation on the survival of human liver cancer cells HepG2. Figure 6 The results show the effect of ALP-D, a derivative of sanshinone, on the survival of mouse liver cancer cells Hepa1-6 and human lung cancer cells H460.

[0051] from Figure 5 It can be seen that after treating human liver cancer cells HepG2 with different concentrations of shanjiangin and shanjiangin derivative ALP-D for 48 hours, the IC50 value decreased significantly. The half inhibitory concentration (IC50) value at 48 hours dropped from 221μM to 61μM. The results showed that the activity of shanjiangin derivative ALP-D increased by 3.6 times that of shanjiangin, and had better killing activity than shanjiangin original drug.

[0052] from Figure 6 It can be seen that after treating human lung cancer cells H460 and mouse liver cancer cells Hepa1-6 with different concentrations of the shanjiangin derivative ALP-D, both showed obvious concentration dependence, that is, as the concentration of the shanjiangin derivative ALP-D increased, the survival rate of tumor cells decreased. The results show that the shanjiangin derivative ALP-D can kill tumor cells from different species.

[0053] Example 3 Cell apoptosis experiment

[0054] The cell apoptosis rate was detected using Annexin V-FITC / 7-AAD-PerCP apoptosis flow cytometry detection kit (Biyuntian, Shanghai, China).

[0055] Human lung cancer cell line H460 and mouse liver cancer cell line Hepa1-6 were inoculated into 6-well plates, with approximately 1×10 5 Tumor cells were treated with different concentrations of the shanjiangsu derivative ALP-D (10 μM and 50 μM, respectively) and an equal amount of solvent control. After 24 h, the cells were treated with Ca-free 2+ / Mg 2+ The cells were washed three times with phosphate buffer solution (PBS, Invitrogen); then digested with 0.05% trypsin (Invitrogen) without EDTA for 5 min, and collected into 1 mL of complete medium to terminate digestion. The cells were centrifuged at 1000 g for 5 min to obtain the cell pellet; then PBS (without Ca) was added. 2+ / Mg 2+) After washing the cell pellet again, resuspend it in 195 μL binding buffer; add 5 μL Annexin V-FITC reagent to the cell suspension, gently vortex and incubate at room temperature in the dark for 30 min; add 5 μL 7-AAD dye and mix gently, and immediately use flow cytometer (BD Biosciences, CA, USA) to analyze cell apoptosis.

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

[0057] Experimental Example 4 Cell migration and clone formation experiment

[0058] Cell migration assay was performed with 10 4 Calculate the number of Hepa1-6 tumor cells and H460 tumor cells, resuspend them in 300μL serum-free cell culture medium, and inoculate them into the upper chamber of the cell migration well. At the same time, add 500μL complete culture medium containing 10% fetal bovine serum (FBS) to the lower chamber; give different concentrations of ALP-D (10 and 50μM) and equal amount of solvent control treatment, and perform cell migration assay 24h later. 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 to stain for 30min, wash with double distilled water and take pictures. The results are attached. Figure 8 .

[0059] In the cell clone formation experiment, 300 Hepa1-6 tumor cells, H460 tumor cells and HepG2 tumor cells were counted in each well and inoculated into 6-well plates or 12-well plates, respectively. The cells were treated with shanjiangin and shanjiangin derivative ALP-D (concentration of 50 μM) and an equal amount of solvent control for 14 days, washed three times with PBS, fixed with cold methanol, stained with 0.4% crystal violet solution for 30 minutes, and washed with double distilled water before taking pictures. The results are shown in the attached Fig. 9 .

[0060] from Figure 8 It can be seen that different concentrations of the alpinia officinalis derivative ALP-D can significantly inhibit the migration ability of H460 tumor cells and Hepa1-6 tumor cells.

[0061] from Fig. 9 It can be seen that compared with the solvent control group, stimulation with a certain concentration of shanjiangin and shanjiangin derivative ALP-D can inhibit the clonal sphere formation of three tumor cells (HepG2, H460 and Hepa1-6), and the inhibitory effect of shanjiangin derivative ALP-D is particularly significant.

[0062] The results showed that the shanjiangin derivative ALP-D could significantly inhibit the formation of clonal spheres of tumor cells from different species, and had a preventive effect on the occurrence of tumors from different species, and its effect was significantly better than that of the shanjiangin original drug.

[0063] Experimental Example 5 Identification of the Binding Target of ALP-D of Alpinia officinalis Derivative

[0064] In this embodiment, the shanjiangsu derivative ALP-D can further introduce a reporter group through a bioorthogonal reaction (click chemistry reaction) by introducing a terminal alkynyl group into the structure. Commonly used reporter groups include two types: the first type is a small molecule fluorescent group, such as rhodamine (Rho) and Cy3 / Cy5, etc. This type of group can be separated by SDS-PAGE and a fluorescent imager can be used to display proteins bound to small molecules; the second type is a biotin group, which is combined with avidin-containing magnetic beads to enzymatically hydrolyze the enriched protein into polypeptides, and then the peptide molecules are identified by high performance liquid chromatography-mass spectrometry (LC-MS / MS). The target protein bound to the drug can be determined based on the characteristic peptides.

[0065] In this example, human liver cancer cells HepG2 were treated with ALP-D, a derivative of shanjiangsu, and an equal amount of solvent control for 4 hours, and then total protein was extracted. Click chemistry reaction was performed according to the protein samples of equal concentrations, and azide-rhodamine (20 μM), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine [Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine, TBTA, 0.1 mM], tris(2-carboxyethyl)phosphine hydrochloride [Tris(2-carboxyethyl)phosphinehydrochloride, TCEP, 1 mM] and copper sulfate (CuSO4, 1 mM) were added in sequence, mixed well, and incubated on a shaker at 37° C. for 3 hours; cold methanol was used for precipitation, the precipitate was washed twice, and 40 μL of 0.4% The protein was dissolved in SDS-PBS, and SDS-PAGE electrophoresis was performed after protein denaturation. The protein labeling was observed using a fluorescent imager, and the PAGE gel was stained with Coomassie Brilliant Blue dye. The results are shown in the attached Fig.10 .

[0066] Similarly, after incubating human liver cancer cells HepG with the shanjiangsu derivative ALP-D and an equal amount of solvent control for 24 hours, the total protein was extracted, and the click chemistry reaction was performed according to the equal concentration of protein samples. Azide-biotin (10μM), TBTA (0.1mM), TCEP (1mM) and CuSO4 (1mM) were added in sequence and mixed well, and then incubated on a shaker at 37°C for 3 hours, precipitated with cold methanol, and washed twice; the protein was dissolved with 1mL of 0.2% SDS-PBS, and 50μL of streptavidin beads were added and rotated at room temperature for 2 hours and then centrifuged; the beads were washed with 1% SDS (3 times), urea (6M, 2 times) and PBS (3 times) in sequence, and DTT reduction, alkylation and enzyme cleavage (25ng / μL trypsin for 16h) were performed. After desalting and freeze-dried peptide samples, MS detection was performed on the machine, and the results were analyzed using the Uniprot human protein database, and the identification results were verified by protein immunoblotting for candidate target enrichment. The results are shown in the attached Fig.11 .

[0067] Fig.10 The results of rhodamine fluorescence imaging are shown, with fluorescent gel on the left and Coomassie brilliant blue staining on the right. The results show that the shanjiangsu derivative ALP-D can enrich the labeled target protein.

[0068] Fig.11 The protein spectrum search results show that the target proteins enriched in the shanjiangsu derivative ALP-D. The left side is a volcano plot showing proteins with a differential expression fold change (FC) greater than two times and a statistical p value less than 0.05; the right side shows six representative target proteins enriched.

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

[0070] Male C57BL / 6 mice weighing approximately 18-22 g were selected, randomly divided into groups, and given sufficient water and food. All operations were performed in an SPF clean environment, and the animal experimental operations were approved by the Ethics Committee of Beijing Shijitan Hospital Affiliated to Capital Medical University (Ethics No. 2023-KY-0009-001).

[0071] Specifically, a cell suspension of mouse liver cancer cells Hepa1-6 (about 10 6 The cells were inoculated subcutaneously in the armpits of mice, and the wounds were sealed with collodion. The mice were then fed normally. After the tumors grew out, the size of the tumors was measured with a vernier caliper. The volume (Tumor Volume, TV) was calculated as follows: TV = (a*b 2 ) / 2, where a is the length of the tumor and b is the width of the tumor. When the tumor grows to about 100 mm 3 At the same time, 20 mg / kg of ALP-D solution of shanjiangsu derivative or the same dose of solvent was intraperitoneally injected as the control group, and the drug was administered three times a week. After 4 weeks of administration, the body weight of each group of mice was weighed and the tumor volume was calculated. After the end of the administration, the mice were anesthetized and killed to obtain the samples, observe and weigh the tumor weight. The results are shown in Table 1 and Attached Fig.12 .

[0072] Table 1 Evaluation results of mouse transplant tumor model

[0073]

[0074]

[0075] Fig.12 The results of evaluating the antitumor activity of the shanjiangsu derivative ALP-D in an in vivo transplant tumor model are shown. Fig.12 It can be clearly seen that after intervention with the shanjiangsu derivative ALP-D, the growth of mouse tumors can be significantly inhibited, and the tumor capsule is obvious and poorly differentiated, indicating that the shanjiangsu derivative ALP-D has significant anti-tumor activity.

[0076] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Those skilled in the art may make some simple modifications, equivalent changes or modifications using the technical contents disclosed above, which all fall within the protection scope of the present invention.

Claims

1. A sanpin derivative, characterized in that The chemical structure of the alpinia officinalis derivative is:

2. The method for preparing the sanpindine derivative according to claim 1, characterized in that: The method comprises the following steps: weighing galangin powder, placing the powder in a reaction bottle, adding 3-bromopropyne, potassium carbonate and dichloromethane, stirring at room temperature, adding water after 12 hours, and extracting the mixture with ethyl acetate; washing the obtained ethyl acetate extract with double distilled water and saturated saline in turn, drying with anhydrous sodium sulfate, separating the extract with a silica gel column, and eluting the extract with a gradient of petroleum ether-ethyl acetate in a volume ratio of 100:1-1:1 to obtain the galangin derivative.

3. The method for preparing the sanpind 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 extraction times of ethyl acetate in the extraction step are at least 3 times.

4. The method for preparing the sanpindine derivative according to claim 3, characterized in that: The yield of the sanpindine derivative obtained by the method is greater than 80%.

5. An anti-tumor drug composition, characterized in that: It comprises the sanpindine derivative or a pharmaceutically acceptable salt thereof as claimed in claim 1, and a pharmaceutically acceptable carrier or excipient.

6. Use of the alpinia officinalis derivative according to claim 1 in the preparation of anti-tumor drugs.

7. The use of the shanjiangsu derivative according to claim 6 in the preparation of anti-tumor drugs, characterized in that: The tumors that can be prevented or treated by the anti-tumor drug include but are not limited to liver cancer and lung cancer.

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