M6A demethylase ALKBH1 small-molecule inhibitor, synthetic method and application

Abstract

The invention discloses an m6A demethylase ALKBH1 small-molecule inhibitor as well as a synthesis method and application thereof, and relates to the technical field of pharmacy, and the m6A demethylase ALKBH1 small-molecule inhibitor has a chemical structural formula as shown in a formula (I). The m6A demethylase ALKBH1 small-molecule inhibitor has biological activity, can be combined with ALKBH1 protein in vitro in a dose-dependent manner, reduces ALKBH1 enzyme activity and has nano-friction-level inhibition activity, and meanwhile, the effect of the small-molecule inhibitor on lung cancer cells A549 is verified, so that the m6A demethylase ALKBH1 small-molecule inhibitor has a good application prospect. A wide application prospect is provided for subsequent computer simulation screening, accurate synthesis of the m6A demethylase ALKBH1 small-molecule inhibitor and application of the m6A demethylase ALKBH1 small-molecule inhibitor in resisting diseases caused by ALKBH1 overexpression. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and more specifically to a small molecule inhibitor of m6A demethylase ALKBH1, a synthesis method and an application thereof. Background Art

[0002] ALKBH1 is a member of the ALKB family. ALKB family proteins are mainly responsible for the methylation modification of DNA and RNA. FTO and ALKBH5 of the ALKB family mainly catalyze the demethylation of DNAm6A, thereby affecting the transcription and translation of downstream genes. Studies have found that ALKBH1 also has the demethylation function of nucleic acids, mainly catalyzing the 3-mec demethylation of ssDNA and RNA. With the continuous deepening of research, it is found that ALKBH1 can regulate tRNA 1mA demethylation and affect the translation process of proteins; ALKBH1 affects the development of astroglioma by regulating the 6mA demethylation of DNA; silencing the ALKBH1 gene can cause DNA and RNAm6A modification, thereby affecting the occurrence and development of cancer. In recent years, the role of ALKBH1 and m6A in lung cancer has been explored in vivo and in vitro. In vivo and in vitro experiments have proved that ALKBH1 regulates the occurrence and development of lung cancer by regulating RNAm6A methylation modification. Silencing the ALKBH1 gene in vitro can inhibit the proliferation, migration, and invasion of lung cancer cells A549; after overexpressing the ALKBH1 gene in vitro, the proliferation, migration, and invasion of lung cancer cells A549 were significantly enhanced. This shows that ALKBH1 does play an important role in the occurrence and development of lung cancer. However, for now, it is necessary to develop small molecule inhibitors of ALKBH1 with biological activity. Summary of the invention

[0003] To solve the above problems, the present invention provides a small molecule inhibitor of m6A demethylase ALKBH1, a synthesis method and application thereof, which can play a role in the treatment of lung cancer.

[0004] The present invention is achieved through the following technical solutions:

[0005] On the one hand, the present invention provides a small molecule inhibitor of m6A demethylase ALKBH1, wherein the small molecule inhibitor of m6A demethylase ALKBH1 has the chemical structure described in formula (I):

[0006]

[0007] In formula (I), the R group is:

[0008]

[0009] On the other hand, the present invention provides a method for synthesizing a small molecule inhibitor of m6A demethylase ALKBH1, which specifically comprises the following steps:

[0010] The target compound required for the R group is weighed, dimethylformamide is added to dissolve it, and potassium carbonate and propargyl bromide solution are added respectively, and the mixture is mixed to obtain a first mixture.

[0011] The ratio of the target compound, dimethylformamide, potassium carbonate and propargyl bromide toluene solution is 366.24mg-368.24mg:8ml-10ml:226mg-228mg:0.36ml-0.38ml.

[0012] The first mixture was stirred in argon for 12 h to 13 h, cooled to room temperature, the solvent was removed, and the intermediate product was purified.

[0013] The intermediate product was weighed, 3′-azido-3′-deoxythymidine, water and n-butanol were added, sodium ascorbate and copper sulfate pentahydrate were added after mixing to obtain a second mixture, the second mixture was stirred in argon for 3 h to 4 h, the liquid was collected, and the solvent was removed to obtain a small molecule inhibitor of 6mA demethylase ALKBH1.

[0014] The ratio of the intermediate product, 3′-azido-3′-deoxythymidine, water and n-butanol solution, sodium ascorbate and copper sulfate pentahydrate is 300mg-302mg: 640.8mg-642.8mg: 4ml-6ml: 38mg-40mg: 10mg-12mg.

[0015] The target compounds are: uracil, adenine, 5-fluorouracil, thymine, 6-azauracil, 5-bromo-6-azauracil, 4-amino-2,6-dihydroxypyrimidine, 5-ethyluracil, and 5-methoxy-2,4-dihydroxypyrimidine.

[0016] Preferably, the volume ratio of propargyl bromide to toluene in the propargyl bromide toluene solution is 4-5:1.

[0017] Preferably, the volume ratio of water to n-butanol in the water and n-butanol solution is 1 to 2:1.

[0018] Preferably, the stirring temperature is 60°C to 65°C.

[0019] Preferably, the m6A demethylase ALKBH1 small molecule inhibitor is used to prepare a therapeutic drug for lung cancer.

[0020] The m6A demethylase ALKBH1 small molecule inhibitor is used to prepare a drug for reducing the proliferation of A549 lung cancer cell line.

[0021] Preferably, the m6A demethylase ALKBH1 small molecule inhibitor is used as a therapeutic drug for treating diseases caused by ALKBH1 overexpression.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention provides a small molecule inhibitor of m6A demethylase ALKBH1. The small molecule inhibitor of m6A demethylase ALKBH1 has biological activity, can bind to ALKBH1 protein in a dose-dependent manner in vitro, reduce ALKBH1 enzyme activity, and has nanomolar inhibitory activity. At the same time, the effect of the small molecule inhibitor on lung cancer cells A549 is verified, which provides broad application prospects for subsequent screening and precise synthesis of small molecule inhibitors of m6A demethylase ALKBH1 through computer simulation and application in combating diseases caused by overexpression of ALKBH1. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0025] Figure 1 Schematic diagram of the chemical structure of MCY-305A, a small molecule inhibitor of m6A demethylase ALKBH1 of the present invention;

[0026] Figure 2 Schematic diagram of the synthesis route of the m6A demethylase ALKBH1 small molecule inhibitor MCY-305A of the present invention;

[0027] Figure 2 In the figure, A is a method for synthesizing an intermediate product; B is a method for synthesizing MCY-305A, a small molecule inhibitor of m6A demethylase ALKBH1;

[0028] Figure 3 It is the nuclear magnetic resonance hydrogen spectrum of the m6A demethylase ALKBH1 small molecule inhibitor MCY-305A of the present invention;

[0029] Figure 4 It is the carbon nuclear magnetic resonance spectrum of the m6A demethylase ALKBH1 small molecule inhibitor MCY-305A of the present invention;

[0030] Figure 5 A schematic diagram of the interaction site between MCY-305A, a small molecule inhibitor of the simulated m6A demethylase ALKBH1, and ALKBH1 obtained by virtual screening in the present invention;

[0031] Figure 6 The present invention uses the MTT method to detect the anti-proliferative effect of MCY-305A on different cell lines of human non-small cell lung cancer cells;

[0032] Figure 6 In the figure, a is the cell proliferation result of lung cancer cell line A549 after 24h treatment with MCY-305A; b is the cell proliferation result of lung cancer cell line A549 after 48h treatment with MCY-305A; c is the cell proliferation result of lung cancer cell line H1299 after 24h treatment with MCY-305A; d is the cell proliferation result of lung cancer cell line H1299 after 48h treatment with MCY-305A, * indicates p<0.1, ** indicates p<0.01, *** indicates p<0.001, **** indicates p<0.0001;

[0033] Figure 7 Schematic diagram of the interaction between the m6A demethylase ALKBH1 small molecule inhibitor MCY-305A and ALKBH1 based on AutoDock simulation of the present invention. DETAILED DESCRIPTION

[0034] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively below, and preferred embodiments of the present invention are provided. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0036] The inventive concept of the present invention is as follows:

[0037] The present invention conducts molecular docking with the chemical structure described in formula (I) and the ALKBH1 protein structure in the PDB database, and finds for the first time that the chemical structure described in formula (I) can be used as a small molecule inhibitor of m6A demethylase ALKBH1. The triazole ring in the small molecule structure is split into an alkynyl group and an azide, and the two can obtain a nitrogen-containing five-membered ring structure in the target small molecule through a cycloaddition reaction. The azide structure is 3'-azido-3'-deoxythymidine, and the alkynyl structure can be obtained by reacting uracil with propargyl bromide. At the same time, according to other similar structures, the group connected to the methylene group connected to the triazole ring can be used as a replacement group, and the rest can be used as its parent core structure to synthesize other similar small molecule compounds, that is, the reactant uracil in the reaction with propargyl bromide is replaced by other target compounds to be synthesized, and then reacted with the azide structure to complete the replacement of the substituent.

[0038] The technical solution of the present invention will be further described below in conjunction with specific embodiments.

[0039] Virtual screening of m6A demethylase ALKBH1 small molecule inhibitors was performed using Discovery Studio Client2019 software, and the compounds were docked with the active sites of the protein structure. First, the ALKBH1 crystal structure was selected from the PDB database, and then the X-RAY DIFFRACTION was less than 1. The ALKBH1 crystal structure, PDB: 6MIC; secondly, the most recent 2016 compound 2D structure database was selected from the National Cancer Institute, NCI, and the Developmental Therapeutics Program, DTP. Finally, molecular docking was performed through the software. First, the protein target was selected: the obtained ALKBH1 crystal structure PDB format file was imported into the Discovery StudioClient 2019 software, the water molecules in the protein structure were deleted, the selected protein was processed using the software "Clean Protein" function, and the active site of the ALKBH1 crystal structure was automatically obtained using the software "Define and EditBinding Site" function, and the ideal site was selected as the target for subsequent docking; secondly, the ligand was prepared: the compound 2D structure database obtained from the NCI-DTP database was imported into the Discovery Studio Client, the target compound was selected, and the software "Prepare Ligands" function was used to prepare the target compound as the ligand required for molecular docking with the protein target; finally, molecular docking was performed and the docking results of the small molecule and the target were obtained: the protein file of the target selected in the early stage was used to carry out molecular docking between the obtained ligand and the processed protein target through the software Dock Ligands function, and the compound with a better score was selected as the result obtained in the early screening according to the docking results. The present invention discovers for the first time that a compound having the chemical structure described in formula (I) can be used as a small molecule inhibitor of m6A demethylase ALKBH1.

[0040]

[0041] In structural formula (I), the R group is:

[0042]

[0043]

[0044] The preparation method of m6A demethylase ALKBH1 small molecule inhibitor is as follows:

[0045] The target compound required for the R group is weighed into a Schlenk tube, dimethylformamide is added to dissolve it, and then potassium carbonate and propargyl benzyl bromide solution are added to the solution, and mixed to obtain a first mixture.

[0046] The ratio of the target compound, dimethylformamide, potassium carbonate and propargyl bromide toluene solution is 366.24mg-368.24mg:8ml-10ml:226mg-228mg:0.36ml-0.38ml.

[0047] The first mixture is stirred in argon, cooled to room temperature, the solvent is removed under reduced pressure, and the desired intermediate product is obtained by separation and purification through column chromatography.

[0048] The intermediate product is weighed and added to a Schlenk tube, 3′-azido-3′-deoxythymidine, water and n-butanol are added, sodium ascorbate and copper sulfate pentahydrate are added, and the second mixture is stirred in argon. After the reaction is completed, the liquid is collected by filtration, and the solvent is removed under reduced pressure. The residue is separated and purified by column chromatography to obtain the desired small molecule compound substituted with different R groups, i.e., a small molecule inhibitor of 6mA demethylase ALKBH1.

[0049] The ratio of the intermediate product, 3′-azido-3′-deoxythymidine, water and n-butanol solution, sodium ascorbate and copper sulfate pentahydrate is 300mg-302mg: 640.8mg-642.8mg: 4ml-6ml: 38mg-40mg: 10mg-12mg.

[0050] The target compounds are: uracil, adenine, 5-fluorouracil, thymine, 6-azauracil, 5-bromo-6-azauracil, 4-amino-2,6-dihydroxypyrimidine, 5-ethyluracil, and 5-methoxy-2,4-dihydroxypyrimidine.

[0051] Example 1. Synthesis of small molecule inhibitors of m6A demethylase ALKBH1

[0052] See also Figure 1 and Figure 2 , for the R group The specific synthesis route is:

[0053] (1) 366.24 mg of uracil (3.27 mmol) was weighed using an electronic balance and placed in a 25 ml Schlenk tube equipped with a magnet. 8 ml of dimethylformamide was added to dissolve the mixture, and then 226 mg of potassium carbonate and 0.36 ml of 80% propargyl bromide toluene solution were added and mixed to obtain a first mixture, wherein the volume ratio of propargyl bromide to toluene was 4:1.

[0054] (2) The first mixture was stirred at 60° C. for 12 h in an argon atmosphere. After the reaction was completed, the mixture was cooled to room temperature, the solvent was removed under reduced pressure, and the intermediate product was obtained by separation and purification by column chromatography. The ratio of petroleum ether to ethyl acetate was 1:1.

[0055] (3) Weigh 2 mmol of the substance by using an electronic balance and separate and purify the intermediate product 300 mg by column chromatography. Add 640.8 mg of 2.4 mmol of 3′-azido-3′-deoxythymidine, water and n-butanol solution, and weigh 4 ml of the mixed solution, wherein the volume ratio of water to n-butanol is 1:1. Add 38 mg of sodium ascorbate (L-Ascorbic Acid Sodium Salt) and 10 mg of copper sulfate pentahydrate (Copper sulfate pentahydrate) to obtain a second mixture, and stir the second mixture at 60°C for 3 h under an argon atmosphere. After the reaction is completed, cool to room temperature, filter and collect the liquid, remove the solvent under reduced pressure, and separate and purify the residue by column chromatography, ethyl acetate: CH3OH = 1:1, to obtain 657 mg of m6A demethylase ALKBH1 small molecule inhibitor, with a yield of 78.8%.

[0056] Example 2: Synthesis of small molecule inhibitors of m6A demethylase ALKBH1

[0057] See also Figure 1 and Figure 2 The specific route for the synthesis of small molecule inhibitors of m6A demethylase ALKBH1 is:

[0058] (1) 368.24 mg of uracil (3.27 mmol) was weighed using an electronic balance and placed in a 25 ml Schlenk tube equipped with a magnet. 10 ml of dimethylformamide was added to dissolve the mixture, followed by 228 mg of potassium carbonate and 0.38 ml of 80% propargyl bromide toluene solution. The mixture was mixed to obtain a first mixture, wherein the volume ratio of propargyl bromide to toluene was 5:1.

[0059] (2) The first mixture was stirred at 65° C. for 13 h in an argon atmosphere. After the reaction was completed, the mixture was cooled to room temperature, the solvent was removed under reduced pressure, and the intermediate product was obtained by separation and purification by column chromatography. The ratio of petroleum ether to ethyl acetate was 1:1.

[0060] (3) An electronic balance was used to weigh 2 mmol of the substance, and 302 mg of the intermediate product was obtained by column chromatography separation and purification. 642.8 mg of 2.4 mmol of 3′-azido-3′-deoxythymidine was added, and 6 ml of the mixed solution was weighed, wherein the volume ratio of water to n-butanol was 2:1. 0.02 mmol, 40 mg of sodium ascorbate, the English name is L-Ascorbic Acid Sodium Salt, and 0.04 mmol, 12 mg of copper sulfate pentahydrate, the English name is Copper sulfate pentahydrate, were added, and mixed to obtain a second mixture. The second mixture was stirred and reacted at 65°C under an argon atmosphere for 3 h. After the reaction was completed, it was cooled to room temperature, the liquid was collected by filtration, and the solvent was removed under reduced pressure. The residue was separated and purified by column chromatography, ethyl acetate: CH3OH = 1:1, and 657 mg of m6A demethylase ALKBH1 small molecule inhibitor was obtained, with a yield of 78.6%.

[0061] The structure of the synthesized compound is confirmed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum, and the nuclear magnetic resonance hydrogen spectrum and carbon spectrum of the compound are obtained, which respectively indicate the number of H and C in the compound, thereby determining the structure. Nuclear magnetic resonance hydrogen spectrum: 1 H NMR (400MHz, DMSO) δ11.37-11.30 (m, 2H), 8.26 (s, 1H), 7.82-7.73 (m, 2H), 6.40 (t, J=6.6Hz, 1H), 5.60 (dd, J=7.8, 1. 9Hz, 1H), 5.40-5.26 (m, 2H), 4.95 (s, 2H), 4.19 (q, J=3.8Hz, 1H), 3.70-3.58 (m, 2H), 2.75-2.59 (m, 2H), 1.80 (s, 3H). Such as Figure 3 As shown, 13 C NMR (101 MHz, DMSO) δ 164.22, 151.23, 150.91, 146.00, 143.16, 136.72, 123.73, 110.12, 101.77, 84.86, 84.32, 61.20, 59.78, 42.89, 37.56, 12.71, one carbon atom signal overlaps, such as Figure 4 shown.

[0062] The molecular formula of the m6A demethylase ALKBH1 small molecule inhibitor is: C 17 H 19 N7O6.

[0063] The chemical structure of the m6A demethylase ALKBH1 small molecule inhibitor is as follows Figure 1 The synthetic route is as shown in Figure 2 As shown in the figure, the m6A demethylase ALKBH1 small molecule inhibitor is named MCY-305A using the abbreviation of the experimenter who synthesized the compound and the sequential number of the experiment performed.

[0064] Example 3: Antiproliferative effect of MCY-305A on human non-small cell lung cancer cell lines

[0065] The MTT assay was used to detect the effect of MCY-305A, a small molecule inhibitor of m6A demethylase ALKBH1, on the proliferation of lung cancer cells A549 and H1299. A549 and H1299 cells in the logarithmic growth phase were taken and 100 μl of cell suspension was added to 5×10 3 / well were inoculated into 96-well plates and cultured in a constant temperature incubator at 37°C and 5% CO2. After the cells adhered to the wall, different concentrations of m6A demethylase ALKBH1 small molecule inhibitor MCY-305A were added for treatment, and 6 duplicate wells were set up in each group. After 24h and 48h of treatment, 150μl of MTT solution with a final concentration of 0.5mg / ml was added to each well and incubated for 4h. After the incubation, the culture medium in the well was discarded, and 150μl of DMSO solution was added to each well. The crystals were completely dissolved by shaking and mixing. The absorbance value of each well at 490nm was measured using an ELISA reader. The effect of different concentrations of the synthetic ALKBH1 small molecule inhibitor MCY-305A on the proliferation of lung cancer cells A549 and H1299 was calculated based on the absorbance value.

[0066] The experimental results are as follows Figure 6 As shown, Figure 6 a in the figure is the cell proliferation result of lung cancer cell line A549 after treatment with MCY-305A for 24 hours; Figure 6 b in the figure shows the cell proliferation results of lung cancer cell line A549 after treatment with MCY-305A for 48 hours; Figure 6 c in the figure is the cell proliferation result of lung cancer cell line H1299 after treatment with MCY-305A for 24 hours; Figure 6 The d in the figure is the cell proliferation result of the lung cancer cell line H1299 after 48 hours of treatment with MCY-305A. The results showed that MCY-305A had anti-proliferative effects on both A54 lung cancer cell lines and H1299 lung cancer cell lines, and had a significant anti-cancer cell proliferation effect on A549 lung cancer cell lines, indicating that the ALKBH1 small molecule inhibitor MCY-305A has biological activity.

[0067] Example 4: Molecular docking of MCY-305A

[0068] First, after MCY-305A was obtained through virtual screening in Discovery Studio, the binding of MCY-305A to ALKBH1 was initially understood through the functions of Discovery Studio software, such as Figure 5 As shown, the interaction sites between the simulated m6A demethylase ALKBH1 small molecule inhibitor MCY-305A and ALKBH1 obtained based on virtual screening. Subsequently, the binding mode of MCY-305A and ALKBH1 was accurately analyzed by AutoDock. The molecular docking analysis of MCY-305A and m6A demethylase ALKBH1 was performed using AutoDock 1.5.7 software. The best binding mode and mode of action of MCY-305A and ALKBH1 were visualized and analyzed using AutoDock 1.5.7 and Pymol. The chemical structure of MCY-305A was processed by AutoDock 1.5.7, and molecular docking was performed with the ALKBH1 protein structure, PDB: 6MIC. The best docking mode was selected from the docking results. The best docking mode was binding energy-7.45kcal / mol. The best docking mode was visualized using Pymol, as shown in the figure. Figure 7 As shown, MCY-305A has one binding site with the amino acid residue LYS-182 of ALKBH1 (PDB: 6MIC), one binding site with VAL-232, one binding site with ARG-234, and three binding sites with SER-235, forming hydrogen bonds. Molecular docking by AutoDock and visualization analysis by Pymol provide a basis for the effect of MCY-305A on ALKBH1, indicating that the amino acid residues obtained by Pymol visualization are the key sites where MCY-305A has pharmacological effects on ALKBH1.

[0069] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0070] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A small molecule inhibitor of m6A demethylase ALKBH1, characterized in that The m6A demethylase ALKBH1 small molecule inhibitor has the chemical structure described in formula (I): In formula (I), the R group is:

2. The method for synthesizing the m6A demethylase ALKBH1 small molecule inhibitor according to claim 1, characterized in that: The specific steps include: Weigh the target compound required for the R group, add dimethylformamide to dissolve it, and then add potassium carbonate and propargyl benzyl bromide solution respectively, mix them, and obtain a first mixture; The ratio of the target compound, dimethylformamide, potassium carbonate and propargyl benzyl bromide solution is 366.24mg-368.24mg:8ml-10ml:226mg-228mg:0.36ml-0.38ml; The first mixture was stirred in argon for 12 h to 13 h, cooled to room temperature, the solvent was removed, and the intermediate product was purified; Weigh the intermediate product, add 3′-azido-3′-deoxythymidine, water and n-butanol, mix, add sodium ascorbate and copper sulfate pentahydrate to obtain a second mixture, stir the second mixture in argon for 3 h to 4 h, collect the liquid, remove the solvent, and obtain a 6mA demethylase ALKBH1 small molecule inhibitor; The ratio of the intermediate product, 3′-azido-3′-deoxythymidine, water and n-butanol solution, sodium ascorbate and copper sulfate pentahydrate is 300 mg to 302 mg: 640.8 mg to 642.8 mg: 4 ml to 6 ml: 38 mg to 40 mg: 10 mg to 12 mg; The target compounds are: uracil, adenine, 5-fluorouracil, thymine, 6-azauracil, 5-bromo-6-azauracil, 4-amino-2,6-dihydroxypyrimidine, 5-ethyluracil, and 5-methoxy-2,4-dihydroxypyrimidine.

3. The method for synthesizing a small molecule inhibitor of m6A demethylase ALKBH1 as claimed in claim 2, characterized in that: The volume ratio of propargyl bromide to toluene in the propargyl bromide toluene solution is 4-5:

1.

4. The method for synthesizing a small molecule inhibitor of m6A demethylase ALKBH1 as claimed in claim 2, characterized in that: The volume ratio of water to n-butanol in the water and n-butanol solution is 1-2:

1.

5. The method for synthesizing a small molecule inhibitor of m6A demethylase ALKBH1 as claimed in claim 2, characterized in that: The stirring temperature is 60°C to 65°C.

6. Use of the m6A demethylase ALKBH1 small molecule inhibitor as claimed in claim 1 in the preparation of therapeutic drugs for lung cancer.

7. Use of the m6A demethylase ALKBH1 small molecule inhibitor as claimed in claim 1 in the preparation of a drug for reducing the proliferation of A549 lung cancer cell line.

8. Use of the m6A demethylase ALKBH1 small molecule inhibitor as claimed in claim 1 in a therapeutic drug for treating diseases caused by overexpression of ALKBH1.

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