Application of compound in preparing medicine for treating allergic rhinitis

Abstract

The present invention discloses the use of a compound in the preparation of a drug for treating allergic rhinitis. The CDK2 inhibitor of the present invention can reduce the inflammatory infiltration of nasal mucosal tissue, effectively improve the degree of inflammation of allergic rhinitis, reduce the secretion of mucus, and has a significant therapeutic effect in treating allergic rhinitis. The synthetic route designed by the present invention involves easy-to-obtain raw materials, simple reaction operation, mild reaction conditions, and can be used for industrial production. The obtained drug has a wide range of applications and conversions and has great practical value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of a compound in preparing a medicine for treating allergic rhinitis. Background Art

[0002] Allergic rhinitis (AR) is a common chronic nasal mucosal disease characterized by hypersensitivity of the nasal mucosa to specific allergens. Typical symptoms of allergic rhinitis include nasal itching, sneezing, runny nose, and nasal congestion, which are more obvious during the pollen season and after exposure to allergens. The pathogenesis of allergic rhinitis is complex and involves the interaction of genetic and environmental factors, among which inflammatory response plays a core role in the onset of allergic rhinitis.

[0003] Allergic rhinitis is a type I hypersensitivity reaction mediated by IgE. It is a chronic inflammatory disease of the nasal mucosa in which active mediators such as histamine, trypsin, leukotrienes and prostaglandins are released mediated by specific IgE after atopic individuals come into contact with allergens, and multiple immune active cells and cytokines participate.

[0004] In allergic rhinitis, a variety of inflammatory cells such as eosinophils, mast cells, and T lymphocytes are involved, and the inflammatory mediators they release, such as leukotrienes, histamine, and cytokines, lead to hyperresponsiveness of the nasal mucosa. In addition, nasal mucosal remodeling is also an important feature of allergic rhinitis, including hyperplasia of nasal mucosal smooth muscle, collagen deposition, and angiogenesis. These changes further aggravate the inflammation and symptoms of the nasal mucosa.

[0005] Currently, traditional treatments for allergic rhinitis mainly rely on antihistamines, nasal corticosteroids (such as budesonide and fluticasone), and leukotriene-removing drugs (such as montelukast). These drugs relieve symptoms by suppressing inflammation and alleviating nasal mucosal symptoms, but long-term use may cause side effects including nasal mucosal dryness, bleeding, and infection. In addition, some patients do not respond well to these treatments, and there is a risk of drug tolerance and side effects. In addition, quick-relief drugs (such as nasal antihistamine sprays) help to quickly relieve acute symptoms, but frequent use may lead to dependence. The limitations and potential side effects of these drugs highlight the need for new treatment strategies.

[0006] Therefore, the development of new drugs, especially those with fewer side effects, better efficacy, and novel mechanisms of action, is crucial to improve the treatment outcomes for patients with allergic rhinitis.

[0007] The cell division cycle is a fundamental process in life, a series of events that occur in cells leading to the formation of two identical daughter cells. CDKs belong to a family of serine / threonine protein kinases that require binding to cell cycle proteins for their kinase activity. Abnormalities in CDKs may cause proliferation, genomic and chromosomal instability, leading to human cancers and promoting cancer progression and invasiveness.

[0008] Cyclin-Dependent Kinase 2 (CDK2) is a member of the CDK family of kinases (CDKs) that regulate the cell cycle. Its inappropriate or excessive activation can lead to uncontrolled cell cycle progression and cause many types of cancer, especially ovarian cancer, uterine cancer, gastric cancer, and cancers associated with amplified CCNE1 genes.

[0009] In our previous study (Pengpeng Niu, et al., Discovery of novel macrocyclic derivatives as potent and selective cyclin-dependent kinase 2 inhibitors, Bioorg Med Chem. 2024 Apr 15:104:117711. doi: 10.1016 / j.bmc.2024.117711. Epub2024 Apr 3.) Inhibitors targeting CDK2 were synthesized. Summary of the invention

[0010] The present invention aims at solving the above-mentioned technical problems in the prior art and provides an application of a compound in preparing a medicine for treating allergic rhinitis.

[0011] The use of a compound in the preparation of a drug for treating allergic rhinitis, wherein the structural formula of the compound is shown in Formula I below:

[0012] ,

[0013] Formula Ⅰ.

[0014] Preferably, the drug can reduce inflammatory infiltration of nasal mucosal tissue, improve the degree of inflammation of allergic rhinitis, and reduce mucus secretion.

[0015] Preferably, the dosage form of the drug is oral, injection or nasal drops.

[0016] Preferably, the compound is prepared by dissolving compound 8 and compound 3 in tetrahydrofuran, a solvent, and reacting them under a H2 atmosphere.

[0017] Compound 8 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazine-4-amine;

[0018] Compound 3 is tert-butyl 3-((allylamino)methyl)azetidine-1-carboxylate.

[0019] More preferably, compound 3 is obtained by reacting compound 1 and compound 2.

[0020] Compound 1 is tert-butyl 3-(aminomethyl)azetidine-1-carboxylate;

[0021] Compound 2 is allyl bromide.

[0022] More preferably, the preparation method of compound 8 is:

[0023] (1) Compound 6 is obtained by reacting compound 4 with compound 5,

[0024] Compound 4 is (2-(allyloxy)-4-(methylsulfonyl)phenyl)methanamine;

[0025] Compound 5 is 4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine;

[0026] Compound 6 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine;

[0027] (2) Compound 6 reacts with PMBCl to obtain compound 7.

[0028] Compound 7 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine;

[0029] (3) Compound 7 is reacted with potassium peroxymonosulfonate to obtain compound 8.

[0030] The present invention further provides a drug for treating allergic rhinitis, the structural formula of the active substance is shown in the following formula I:

[0031] ,

[0032] Formula Ⅰ.

[0033] Preferably, the dosage form of the drug is oral, injection or nasal drops.

[0034] The present invention successfully discovered a new macrocyclic compound CDK2 inhibitor with significant anti-inflammatory activity, which can significantly alleviate the nasal mucosal reactivity and inflammatory response of allergic rhinitis.

[0035] The CDK2 inhibitor of the present invention can reduce the inflammatory infiltration of nasal mucosal tissue, effectively improve the degree of inflammation of allergic rhinitis, reduce the secretion of mucus, and has a significant therapeutic effect in treating allergic rhinitis.

[0036] The present invention optimizes the chemical structure of small molecule drugs and improves the metabolic stability of the compounds by introducing cyclobutylamine groups, so that the drugs will not be rapidly metabolized in the body and can maintain effective concentrations in the body for a long time, thereby enhancing the therapeutic effect of the drugs.

[0037] The synthetic route designed by the present invention involves readily available raw materials, simple reaction operation, mild reaction conditions, can be used for industrial production, and the obtained medicine has wide application and transformation and great practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is the reaction formula for the synthesis of CDK2 inhibitors.

[0039] Figure 2 This is a statistical analysis chart of the number of sneezing times observed in mice for 15 minutes after the modeling of each group of mice in the embodiment of the present invention.

[0040] Figure 3 This is a statistical analysis chart of the number of times the mice scratched their noses for 15 minutes after the modeling of each group of mice in the embodiment of the present invention was completed.

[0041] Figure 4 This is a statistical analysis chart of the runny nose scores of mice observed 15 minutes after the modeling of each group of mice in the embodiment of the present invention.

[0042] Figure 5 The white blood cells and their classification count statistics in the bronchoalveolar lavage fluid of each group of mice in the embodiment of the present invention are shown in FIG. Figure 5 A is the total number of white blood cells, Figure 5 B in the figure is the number of eosinophils. Figure 5 C in is the number of neutrophils, Figure 5 D in is the number of lymphocytes, Figure 5 E in the figure is the number of macrophages.

[0043] Figure 6 Statistical graph of histamine in the serum of each group of mice in the examples of the present invention.

[0044] Figure 7 This is a statistical analysis chart of immunoglobulin E (IgE) in the serum of each group of mice in the examples of the present invention.

[0045] Figure 8 Statistical analysis of various cytokines in the serum of each group of mice in the embodiment of the present invention, wherein: Figure 8 A in the equation is the content of interleukin-4 (IL-4). Figure 8 B in the figure is the content of interleukin 5 (IL-5). Figure 8 C in the figure is the content of interleukin-6 (IL-6). Figure 8 D in is the content of interleukin 13 (IL-13), Figure 8 E in it represents the content of interleukin 17 (IL-17).

[0046] Fig. 9 Representative images of HE staining and AB-PAS staining of nasal mucosal tissues of mice in each group to show that CDK2 inhibitor improves lung pathological changes in mice with allergic rhinitis. DETAILED DESCRIPTION

[0047] Statistical analysis in the present invention:

[0048] The measurement data were expressed as mean ± standard deviation (mean ± SD). Multiple groups of measurement data were compared by one-way analysis of variance (ANOVA). Statistical analysis was performed using GraphPad Prism 9.5 software. P < 0.05 was considered to be statistically significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ns indicated no statistical difference.

[0049] Example 1

[0050] A method for synthesizing a macrocyclic compound (CDK2 inhibitor in this application), in the first step, N-Boc-1,2-diethyl-3-propenylamine is generated by using N-Boc-protected diethylamine as a starting material, 3-bromopropylene as an alkylating agent, triethylamine as a base, and dichloromethane as a solvent under appropriate reaction conditions. In the second step, 4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine and 2-(allyloxy)-4-(methylsulfonyl)benzaldehyde are mixed, and the amino group is protected with p-methoxybenzyl chloride (PMBCl) through a series of substitution and oxidation reactions to obtain the second step product. Finally, the products of the first and second steps are mixed, and the final product is obtained through substitution, catalytic hydrogenation and other reactions.

[0051] The reaction formula of the synthesis method is as follows Figure 1 As shown, the specific steps of the synthesis method are:

[0052] Step 1: tert-butyl 3-(aminomethyl)azetidine-1-carboxylate (2.14 g, 10.0 mmol, 1.0 equiv, recorded as compound 1) and TEA (triethanolamine, 4.16 ml, 30.0 mmol, 3.0 equiv) were added to anhydrous DCM (dichloromethane, 50 ml) solution at room temperature, and allyl bromide (0.86 ml, 10.0 mmol, 1.0 equiv, recorded as compound 2). The reaction mixture was stirred at room temperature for 12 h. After the reaction was completed, the product was purified by silica gel column chromatography (5%-10% MEOH / DCM) to obtain a colorless liquid, namely tert-butyl 3-((allylamino)methyl)azetidine-1-carboxylate (recorded as compound 3).

[0053] Step 2: DIPEA (N,N-diisopropylethylamine, 0.3 mL, 1.5 mmol, 1.5 equiv) was added to anhydrous DCM (5 ml) and the intermediate 4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine (242.1 mg, 1.0 mmol, 1.0 equiv, recorded as compound 5) and (2-(allyloxy)-4-(methylsulfonyl)phenyl)methanamine (255.1 mg, 1.0 mmol, 1.0 equiv, recorded as compound 4) were mixed. The reaction mixture was then stirred at room temperature for 1 h and diluted with ethyl acetate and water after the reaction was completed. The organic layer was washed with brine, dried over anhydrous Na2SO4 and removed in vacuo. The resulting residue was subjected to silica gel column chromatography on silica gel (10%-30% EA / PE (ethyl acetate / petroleum ether)) to obtain the intermediate product N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine (322.7 mg, yield 70%, recorded as compound 6) as a white solid.

[0054] Then, a mixture of N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine (461.2 mg, 1.0 mmol, 1.0 equiv) and PMBCl (0.3 mL, 2.0 mmol, 2.0 equiv) was added in anhydrous DMF (N,N-dimethylformamide, 5 mL). The reaction mixture was then stirred at 60 °C for 3 hours and after completion of the reaction, it was diluted with ethyl acetate and water. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and removed in vacuo. The resulting residue was subjected to column chromatography on silica gel (10%-30% EA / PE) to obtain the intermediate N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine (464.8 mg, 80%, recorded as compound 7) as a white solid.

[0055] A mixture of intermediate N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine (581.2 mg, 1.0 mmol, 1.0 equiv) and Oxone (potassium peroxymonosulfonate, 3.1 g, 5.0 mmol, 5.0 equiv) was added to the solution (MeCN / H2O=2:1, 10 mL). The reaction mixture was then stirred at room temperature for 12 h and diluted with ethyl acetate and water after the reaction was completed. The organic layer was washed with brine, dried over anhydrous Na2SO4, and removed in vacuo. The resulting residue was subjected to column chromatography on silica gel (20%-50% EA / PE) to obtain the intermediate N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazine-4-amine (429.1 mg, yield 70%, recorded as compound 8) as a white solid.

[0056] Step 3: The intermediate N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazine-4-amine and tert-butyl 3-((allylamino)methyl)azetidine-1-carboxylate were dissolved in THF (tetrahydrofuran, 5.0 mL) and hydrogenated under 10% Pd / C (palladium / carbon) under H2 atmosphere (balloon) at room temperature for 12 h. After the reaction was completed, the reaction mixture was filtered to remove THF. The crude material dissolved in TFA (trifluoroacetic acid) was stirred at room temperature for 12 h. After the reaction was completed, it was diluted with DCM and washed with saturated aqueous NaHCO3. The organic layer was washed with brine, dried over anhydrous Na2SO4 and removed in vacuo. The resulting residue was subjected to preliminary HPLC to obtain the desired product (Z)-11-(3-azetidinylmethyl)-18-isopropyl-44-(methylsulfonyl)-5-oxo-2,11-diaza-1(4,2)-pyrazolo[1,5-a][1,3,5]triazine-4(1,2)-benzocycloundecane (referred to as compound 9).

[0057] Example 2

[0058] Establishment of mouse model of allergic rhinitis.

[0059] Establishment of mouse model of allergic rhinitis and intervention method of CDK2 inhibitor:

[0060] 6-8 week old male SPF BALB / c mice (n=20), weighing 20-30 g, were randomly divided into 4 groups: Ⅰ normal control group (Ctrl group), Ⅱ allergic rhinitis group (OVA group), Ⅲ CDK2i treatment group (OVA+CDK2i group) and Ⅳ dexamethasone inhibition group (OVA+DEX group) (5 mice in each group, n=5). Groups Ⅱ-Ⅳ were sensitized by intraperitoneal injection of 20 μg ovalbumin (OVA) + 0.5 mg Al(OH)3 + 100 μL PBS on days 0, 7 and 14, respectively, and the normal group was given an equal volume of PBS.

[0061] After injection, from the 21st day to the 27th day, the sensitized mice were subjected to bilateral nasal challenge with 25 mg / ml OVA challenge reagent (50 μL micropipette nasal administration), once a day on each side, 20 μL / time (i.e. 500 μg OVA / time), and the normal control group mice were subjected to sensitization and challenge experiments using PBS as a placebo instead of OVA. Rhinitis mice showed positive characteristics of successful challenge such as restlessness, scratching nose, sneezing, and runny nose during the challenge process. OVA+CDK2i group mice were given oral administration of 0.1 mg / ml CDK2 inhibitor (the product prepared in Example 1) 100 μL / 10g, i.e. 2 mg / kg per day; OVA+DEX group mice were given intraperitoneal injection of 0.2 mg / ml dexamethasone 100 μL / 10g body weight every day from the 21st day, i.e. 2 mg / kg per day, and the other groups were given the same PBS.

[0062] After the last OVA instillation, the symptoms of the mice, such as nose scratching, sneezing, and runny nose, were recorded within 15 minutes. The scoring criteria were as follows: 1 point for nose scratching ≤2 times per minute, 2 points for mild nose scratching (greater than 2 times and less than or equal to 5 times), and 3 points for severe nose scratching (greater than 5 times); 1 point for sneezes 1-3 times / min, 2 points for 4-10 times / min, and 3 points for more than 11 times / min; 1 point for clear mucus flowing to the anterior nares, 2 points for exceeding the anterior nares, and 3 points for clear mucus covering the face; the total score of the sum of the scores of each symptom was greater than or equal to 5 points, indicating that the model was successfully established.

[0063] The symptoms of each group of mice after nasal drip were recorded to evaluate the severity of allergic rhinitis in mice. The results are shown in Table 1 and Figures 2~4 .

[0064] Table 1

[0065]

[0066] The results showed that compared with the normal control group, the total score of mice in the allergic rhinitis model group increased significantly, reaching five points, and the model was successfully established. And after the mice were treated with CDK2 inhibitors or dexamethasone, their scores decreased compared with the allergic rhinitis group. The above results show that CDK2 inhibitors and dexamethasone have similar effects in relieving inflammation, and can relieve the behaviors of itching, scratching, sneezing and runny nose in mice with allergic rhinitis caused by OVA.

[0067] Example 3

[0068] CDK2 inhibitor alters bronchoalveolar lavage fluid leukocyte differential counts in mice with allergic rhinitis.

[0069] Methods: After anesthetizing mice, endotracheal intubation was performed for bronchoalveolar lavage, which was performed 3 times with cold PBS (1 ml / time), and the bronchoalveolar lavage fluid (BALF) was fully recovered and collected. The total number of white blood cells was counted under a microscope using a blood cell counting plate. Smears were made, and after Swiss-Giemsa staining, cell classification and counting were performed under a microscope. 200 white blood cells were counted for each specimen, and eosinophils, neutrophils, lymphocytes, and macrophages were counted separately to obtain the percentage of each type of cell and calculate the absolute value.

[0070] Results: The leukocytes of each group of mice were analyzed and counted (see Figure 5 ), the total number of leukocytes and various types of leukocytes in the allergic rhinitis group were significantly higher than those in the control group, and the number of leukocytes was significantly reduced after treatment with CDK2 inhibitor or dexamethasone. Moreover, CDK2 inhibitor reduced eosinophils to a greater extent than dexamethasone.

[0071] Example 4

[0072] To examine the effects of CDK2 inhibitors on histamine and IgE in allergic rhinitis.

[0073] Methods: 24 hours after the last OVA challenge, blood samples were collected from the mouse eye sockets and centrifuged at 4°C, 2000×g for 10 min to obtain serum. Histamine in the blood supernatant was detected using an enzyme-linked immunosorbent assay kit (see Figure 6 ) and IgE (see Figure 7 ) content.

[0074] Results: In this experiment, compared with the control group, the levels of histamine and IgE in the serum of the mice in the model group after OVA sensitization were significantly increased, and the levels of histamine and IgE in the serum of the mice treated with CDK2 inhibitors showed a downward trend, and the degree of decrease was stronger than that of dexamethasone, which proved the potential of CDK2 inhibitors in the treatment of allergic rhinitis.

[0075] Example 5

[0076] To examine the effects of CDK2 inhibitors on inflammatory factors in allergic rhinitis.

[0077] Methods: 24 hours after the last OVA challenge, blood samples were collected from the mouse orbits and centrifuged at 4°C, 2000×g for 10 min to obtain serum. The levels of cytokines such as IL-4, IL-5, IL-6, IL-13, and IL-17 in the blood supernatant were detected using an enzyme-linked immunosorbent assay kit.

[0078] Results: Figure 8As shown, it was found that the levels of various cytokines in the OVA group mice were higher than those in the Ctrl group, and after intervention with CDK2 inhibitors, their levels were reduced, which was consistent with the trend of the dexamethasone inhibition group.

[0079] Example 6

[0080] CDK2 inhibitor alters lung pathological changes in mice with allergic rhinitis.

[0081] Methods: The nasal mucosal tissues of mice were fixed in 4% paraformaldehyde for 24-48 hours, embedded in paraffin and sliced. The slices were stained with hematoxylin-eosin (HE) and Alcian blue-periodic acid-Schiff (AB-PAS) and observed under an optical microscope to analyze the changes in airway inflammation, lung tissue structure, inflammatory cell infiltration, mucus secretion and collagen deposition.

[0082] Results: HE staining results (see Fig. 9 ) showed that the pseudostratified ciliated columnar epithelial structure of the nasal mucosa of mice in the control group was intact, the cilia were neatly arranged and dense, smooth and without breaks, and there was less inflammatory cell infiltration; in the OVA group, the nasal mucosa had reduced cilia, goblet cells proliferated, the basal layer thickened, inflammatory cell infiltration was observed, and the submucosal small blood vessels were thickened, showing typical characteristics of allergic rhinitis; the CDK2i-treated group and the dexamethasone-treated group could significantly alleviate the nasal mucosal lesions, among which the CDK2i-treated group had a better effect than the dexamethasone-treated group.

[0083] Goblet cells have the function of secreting mucus. When the nasal mucosa is damaged, goblet cells proliferate. Fig. 9 ), goblet cells appear blue. The goblet cell secretions of the nasal mucosa of mice in the control group were purple-red; the goblet cells of the nasal mucosa of mice in the OVA group increased and appeared blue; the goblet cells in the CDK2i and dexamethasone groups were significantly reduced, indicating that the symptoms of allergic rhinitis were significantly improved, and the effect of the CDK2i group was better than that of the dexamethasone group.

[0084] In summary, CDK2 inhibitors can significantly regulate multiple inflammatory indicators in mice with allergic rhinitis. In addition, CDK2 inhibitors have good effects in inhibiting inflammatory cell infiltration in nasal mucosal tissue and goblet cell mucus secretion. This shows that CDK2 inhibitors have excellent therapeutic effects on allergic rhinitis.

Claims

1. Use of a compound in the preparation of a medicament for treating allergic rhinitis, characterized in that: The structural formula of the compound is shown in Formula I below: ， Formula Ⅰ.

2. The use according to claim 1, characterized in that: The drug can reduce the inflammatory infiltration of nasal mucosal tissue, improve the inflammation degree of allergic rhinitis, and reduce the secretion of mucus.

3. The use according to claim 1, characterized in that: The dosage form of the medicine is oral preparation, injection or nasal drops.

4. The use according to claim 1, characterized in that: The compound is prepared by dissolving compound 8 and compound 3 in tetrahydrofuran, a solvent, and reacting them under a H2 atmosphere. Compound 8 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazine-4-amine; Compound 3 is tert-butyl 3-((allylamino)methyl)azetidine-1-carboxylate.

5. The use according to claim 4, characterized in that: Compound 3 is obtained by reacting compound 1 and compound 2. Compound 1 is tert-butyl 3-(aminomethyl)azetidine-1-carboxylate; Compound 2 is allyl bromide.

6. The use according to claim 4, characterized in that: The preparation method of compound 8 is: (1) Compound 6 is obtained by reacting compound 4 with compound 5, Compound 4 is (2-(allyloxy)-4-(methylsulfonyl)phenyl)methanamine; Compound 5 is 4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine; Compound 6 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine; (2) Compound 6 reacts with PMBCl to obtain compound 7. Compound 7 is N-(2-(but-3-en-1-yloxy)-4-(methylsulfonyl)benzyl)-8-isopropyl-N-(4-methoxybenzyl)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-4-amine; (3) Compound 7 is reacted with potassium peroxymonosulfonate to obtain compound 8.

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