A uracil derivative, and a preparation method and application thereof

Abstract

The present application relates to the field of medicine, and particularly relates to a kind of uracil compound containing formula (I) and its application in preparing bacterial virulence factor lasB inhibitor.The uracil compound involved in the present application is verified by in vitro drug efficacy, and most of the obtained compounds have excellent lasB inhibitory activity, can be used for preparing anti-toxicity agent, and has the potential to be developed as a new antibacterial drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a preparation method of a uracil derivative and application thereof in preparation of a bacterial biofilm inhibitor. BACKGROUND

[0002] Long-term use of antibiotics for antibacterial treatment leads to antibiotic resistance and "superbugs" to appear widely worldwide. At present, 7 million people die of microbial drug-resistant infections worldwide each year, and the problem of bacterial drug resistance has constituted a global major public health threat. Pseudomonas aeruginosa is a widely used gram-negative opportunistic pathogen associated with nosocomial infections, which has high invasiveness and pathogenicity. The emergence of bacterial drug resistance makes Pseudomonas aeruginosa infection difficult to treat, and the production of elastase, exotoxin, biofilm and other virulence factors exacerbates the severity of Pseudomonas aeruginosa infection. Pseudomonas aeruginosa has been listed as one of the ESKAPE pathogens by the World Health Organization (WHO). Therefore, it is imminent to develop new treatment strategies for multi-drug resistant Pseudomonas aeruginosa. The production of virulence factors and the formation of biofilm are the main causes of pathogenicity of Pseudomonas aeruginosa, and these are regulated by the quorum sensing (QS) system. There are mainly three QS systems in Pseudomonas aeruginosa, las, rhl and pqs. Among them, the las system occupies a core position, and the signal molecule N-(3-oxododecanoyl)-L-homoserine lactone (3OC12-HSL) produced by the las system binds to the receptor protein LasR, which activates the expression of rhlI, rhlR, pqsR and pqsABCDE genes. LasR can directly regulate the production of elastase lasB, alkaline protease and exotoxin and other virulence factors, and regulate the production of rhamnolipid, pyocyanin and biofilm through regulating rhl and pqs systems. Studies have shown that targeting lasB can reduce the pathogenicity of Pseudomonas aeruginosa, so developing new inhibitors of virulence factor lasB regulated by the las system is an effective strategy for treating Pseudomonas aeruginosa infection. SUMMARY

[0003] The present application aims to at least solve one of the technical problems existing in the prior art. To this end, the present application provides a uracil compound containing a structure shown in formula (I) and a preparation method and use thereof, which can selectively act on Pseudomonas aeruginosa and inhibit the virulence factor lasB regulated by the las quorum sensing system of Pseudomonas aeruginosa.

[0004] In a first aspect of the present application, a compound shown in formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof is provided:

[0005]

[0006] In a second aspect of the present application, a pharmaceutical composition is provided, comprising at least one of the compound of formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, and optionally a pharmaceutically acceptable excipient or carrier.

[0007] The compound of the present application, or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, or the pharmaceutical composition can be administered in unit dosage form, and the route of administration can be oral, intramuscular, subcutaneous, nasal, buccal, dermal, peritoneal or rectal. The administration dosage form can be tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal preparations, buccal tablets, suppositories, or lyophilized powder injections. It can be a general preparation, a sustained-release preparation, a controlled-release preparation, and various microparticle drug delivery systems.

[0008] Generally, the pharmaceutical composition of the present application contains 0.1-99.9% of the compound, or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, by mass percentage.

[0009] In some embodiments of the present application, the administration form of the pharmaceutical composition comprises simultaneous, separate or sequential administration of the compound, or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, and other active pharmaceutical ingredients.

[0010] In a fourth aspect of the present application, a combination drug is provided, comprising at least one of the compound of formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, and an antibiotic.

[0011] Preferably, the antibiotic comprises ciprofloxacin, tobramycin.

[0012] Preferably, the mass ratio of the compound of formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof to the antibiotic is 1:0.5-10, preferably 1:2-3, and more preferably 1:2-2.5.

[0013] In a fourth aspect of the present application, the compound of formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, or the pharmaceutical composition described above is used for preparing a bacterial lasB inhibitor.

[0014] Preferably, the bacteria is Pseudomonas aeruginosa.

[0015] The present application also provides the use of the compound of formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof, or the pharmaceutical composition described above for preparing an antibacterial drug.

[0016] Preferably, the bacteria is Pseudomonas aeruginosa.

[0017] The compound of formula (I) of the present application, especially the compound (R)-6-(3-aminopyrrolidin-1-yl)-3-(benzo[d]thiazol-2-ylmethyl)-1-(but-2-yn-1-yl)pyrimidine-2,4(1H,3H)-dione, has excellent lasB inhibitory activity.

[0018] The uracil derivative of formula (I) of the present application has the chemical structure as shown in Table 1:

[0019] Table 1 Structure and name of compound

[0020]

[0021]

[0022]

[0023] The term "pharmaceutically acceptable salt" in the present application includes a conventional salt formed with a pharmaceutically acceptable inorganic or organic acid, or inorganic or organic base.

[0024] The "pharmaceutical composition" includes a product containing a therapeutically effective amount of the compound of the present application, and any product directly or indirectly resulting from the combination of the compounds of the present application.

[0025] The fourth aspect of the present application provides a preparation method of the uracil derivative of formula (I), which specifically comprises the following steps:

[0026] Route one comprises the following steps:

[0027] (1.1) The starting material 6-chlorouracil 1 reacts with 1-bromo-2-butyn to obtain the intermediate 2;

[0028] (1.2) The intermediate 2 reacts with a bromine-substituted compound through an alkylation reaction to obtain the intermediate 3a-3l;

[0029] (1.3) The intermediate 3a-3l reacts with (R)-3-aminopiperidine dihydrochloride to obtain the final product 4a-4l;

[0030] The synthesis route at this time is:

[0031]

[0032] Route two comprises the following steps:

[0033] (2.1) The intermediate 2 reacts with methyl 3-(bromomethyl)benzoate through an alkylation reaction to obtain the intermediate 4;

[0034] (2.2) The intermediate 4 reacts with (R)-3-Boc-aminopiperidine to obtain the intermediate 5;

[0035] (2.3) Intermediate 5 is further hydrolyzed to obtain intermediate 6;

[0036] (2.4) Intermediate 6 is reacted with different amino-substituted compounds 7a-7m to obtain intermediates 8a-8m;

[0037] (2.5) Intermediates 8a-8m are hydrolyzed to remove the Boc protecting group to obtain target products 9a-9m;

[0038]

[0039] Scheme three, comprising the following steps:

[0040] (3.1) Intermediate 2 is added to a DMF solution, sodium hydride and lithium bromide are added under ice bath conditions, and then a bromo-substituted compound is added to the reaction solution after stirring, and the reaction is carried out at room temperature overnight; post-processing: water is added to the reaction solution to terminate the reaction, extraction is carried out with ethyl acetate, the organic phases are combined, washed with water, dried with anhydrous sodium sulfate, and then purified by a silica gel chromatographic column to obtain intermediates 3r and 3p;

[0041] (3.2) Intermediates 3r or 3p, substituted amine, sodium bicarbonate and activated molecular sieve (4A) are added to an ethanol solution, and the reaction is carried out at 100°C for 2.5h; post-processing: the reaction is concentrated and purified by a silica gel chromatographic column to obtain end products 10a-10f;

[0042]

[0043] The compound according to the embodiment of the present application has at least the following beneficial effects:

[0044] (1) The present application provides a uracil compound containing a structure shown in formula (I), wherein compound 10e has excellent bacterial lasB inhibitory activity.

[0045] (2) The preparation method of the compound shown in formula (I) has the advantages of short route, high yield, convenient post-processing, and good economy. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1Synergistic effect of compound 10e on CIP. (A) Bacterial plating results of the infected wound area of mice in Control group, single compound group, single CIP administration group and 10e-CIP combined administration group. Each group was repeated for 3 times. (B) Monitoring of the wound area of mice, and the wound area was calculated using Image J. (C) Photographic monitoring of the infected wound area of mice. (D) Bacterial survival rate of the infected wound area of mice. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (E) H&E staining slice images of the heart, liver, spleen, lung and kidney of mice in each group.

[0047] Figure 2 Synergistic effect of compound 10e on Tob. (A) Bacterial plating results of the infected wound area of mice in Control group, single Tob administration group and 10e-Tob combined administration group. Each group was repeated for 3 times. (B) Monitoring of the wound area of mice, and the wound area was calculated using Image J. (C) Photographic monitoring of the infected wound area of mice. (D) Bacterial survival rate of the infected wound area of mice. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (E) H&E staining slice images of the heart, liver, spleen, lung and kidney of mice in each group. DETAILED DESCRIPTION

[0048] The application will be further described in conjunction with the specific embodiments.

[0049] Example 1: Preparation of (R)-2-(4-(3-aminopiperidin-1-yl)-3-(but-2-yn-1-yl)-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)acetonitrile (4a).

[0050] To 50.00 ml of N,N-dimethylformamide (DMF) solution, 6-chloropyrimidine-2,4(1H,3H)-dione (29.30 g, 0.20 mol) and N,N-diisopropylethylamine (DIPEA) (35.00 ml, 0.30 mol) were added, and stirred at room temperature for 15 min. Then, 1-bromo-2-butyn (19.60 ml, 0.22 mol) was added to the reaction solution, and stirred at room temperature overnight. Post-treatment: a large amount of water was added to the reaction solution, and a light yellow flocculent precipitate was precipitated, which was filtered under suction, washed with petroleum ether and dried to obtain a light yellow powder, which was intermediate 2 (35.60 g, yield 90%).

[0051] To a solution of 10.00 ml DMF was added intermediate 2 (1.40 g, 7.00 mmol), sodium hydride (NaH) (0.34 g, 14.00 mmol) and lithium bromide (LiBr) (1.80 g, 21.00 mmol) was added under ice bath condition and stirred for 20 min. Then different commercially available bromo substituted compounds (10.50 mmol) was added to the reaction mixture and stirred at room temperature overnight. Work-up: the reaction was quenched by the addition of water and extracted with ethyl acetate, the organic phases were combined, washed with water and dried over anhydrous sodium sulfate for 2 h, then purified by silica gel chromatography to give the crude product intermediate 3a-3l (yield 60-95%).

[0052] To a solution of 20.0 ml ethanol was added intermediate 3a (2.50 mmol), (R)-3- aminopiperidine dihydrochloride (0.65 g, 3.75 mmol), sodium bicarbonate (1.05 g, 12.50 mmol) and 100.00 mg activated molecular sieves (4A) respectively, and stirred at 100 °C for 2.5 h. Work-up: the reaction was concentrated and purified by silica gel chromatography to give the product 4a as a white solid in 57% yield; purity: 95.98%. 1 H NMR (400 MHz, Chloroform-d6) δ 5.24 (s, 1H), 4.79 (s, 2H), 4.54 (t, J = 2.4 Hz, 2H), 3.38 - 3.31 (m, 1H), 3.24 (t, J = 7.4 Hz, 1H), 3.07 - 2.98 (m, 1H), 2.78 - 2.68 (m, 1H), 2.57 - 2.48 (m, 1H), 2.03 - 1.94 (m, 1H), 1.92 - 1.85 (m, 1H), 1.82 (t, J = 2.4 Hz, 3H), 1.74 - 1.62 (m, 1H), 1.55 (s, 2H), 1.36 - 1.22 (m, 1H). 13 C NMR (100 MHz, Chloroform-d6) δ 161.00, 160.20, 151.20, 114.68, 88.28, 80.94, 73.16, 59.18, 51.42, 47.45, 36.07, 33.11, 28.23, 23.17, 3.64. HRMS (ESI) m / z: calcd for C 15 H 20 N5O2 + [M+H] + : 302.1617, found: 302.1622.

[0053] Example 2: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3- (thiophen-3-ylmethyl)pyrimidine-2,4(lH,3H)-dione (4b).

[0054] Using intermediate 2 and 3-(bromomethyl)thiophene, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method is the same as that of compound 4a, the product 4b is collected as a yellow solid in a yield of 60% and a purity of 95.00%. 1 H NMR (400 MHz, Chloroform-d6) δ 7.40 (s, 1H), 7.23 (d, J = 5.0 Hz, 1H), 7.20 (d, J = 3.4 Hz, 1H), 5.21 (s, 1H), 5.07 (s, 2H), 4.53 (q, J = 2.4 Hz, 2H), 3.31 (d, J = 11.7 Hz, 1H), 3.19 (d, J = 12.0 Hz, 1H), 3.01 (tt, J = 8.8, 4.0 Hz, 1H), 2.68 (t, J = 11.2 Hz, 1H), 2.48 (t, J = 10.2 Hz, 1H), 2.05 - 1.94 (m, 2H), 1.82 (t, J = 2.3 Hz, 3H), 1.76 (s, 1H), 1.65 (dd, J = 13.9, 10.3 Hz, 1H), 1.27 (dt, J = 14.3, 8.4 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 162.07, 159.85, 152.06, 138.23, 128.43, 126.51, 123.79, 88.07 (2C), 75.08, 59.12, 51.24, 47.66, 39.28, 35.93, 33.24, 23.39, 3.55. HRMS (ESI) m / z: calcd for C 18 H 23 N4O2S + [M+H] + : 359.1542, found: 359.1557.

[0055] Example 3: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3- (thiophen-2-ylmethyl)pyrimidine-2,4(lH,3H)-dione (4c).

[0056] Using intermediate 2 and 2-(bromomethyl)thiophene, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method is the same as that of compound 4a, the product 4c is collected as a yellow solid in a yield of 40% and a purity of 95.01%. 1H NMR (400 MHz, Chloroform-d6) δ 7.23 - 7.19, 6.95 - 6.91 (m, 1H), 6.96 (d, J = 3.7 Hz, 1H), 6.87 (d, J = 3.7 Hz, 1H), 5.23 (s, 1H), 5.16 (s, 2H), 4.55 - 4.52 (m, 2H), 3.37 - 3.29 (m, 1H), 3.22 (d, J = 11.8 Hz, 1H), 3.03 (tt, J = 8.6, 3.8 Hz, 1H), 2.72 (d, J = 11.5 Hz, 1H), 2.50 (s, 1H), 2.03 - 1.95 (m, 1H), 1.84 (t, J = 2.4 Hz, 3H), 1.73 - 1.64 (m, 1H), 1.54 (s, 2H), 1.39 - 1.22 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 161.79, 160.02, 151.83, 141.23, 129.91, 129.18, 111.73, 87.97, 80.21, 74.94, 58.85, 51.25, 47.56, 39.00, 36.04, 33.01, 23.29, 3.57. HRMS (ESI) m / z: calcd for C 18 H 23 N4O2S + [M+H] + : 359.1542, found: 359.1541.

[0057] Example 4: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3-((5- chlorothiophen-2-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4d).

[0058] Using intermediate 2 and 2-(bromomethyl)-5-chlorothiophene, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method is the same as that of compound 4a, the product 4d is collected as a yellow solid, yield: 30%, purity: 95.06%. 1H NMR (400 MHz, Chloroform-d6) δ 6.99 (d, J = 3.7 Hz, 1H), 6.74 (d, J = 3.7 Hz, 1H), 5.24 (s, 1H), 5.14 (s, 2H), 4.55 (d, J = 1.3 Hz, 2H), 3.34 (d, J = 11.6 Hz, 1H), 3.23 (d, J = 12.0 Hz, 1H), 3.09 - 3.00 (m, 1H), 2.72 (t, J = 11.0 Hz, 1H), 2.52 (s, 1H), 2.05 - 1.96 (m, 1H), 1.85 (t, J = 2.3 Hz, 3H), 1.65 - 1.59 (m, 2H), 1.38 - 1.27 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 161.81, 160.05, 151.84, 138.47, 128.45, 128.24, 126.35, 87.92, 80.21, 79.63, 74.91, 59.18, 51.22, 47.64, 36.03, 33.29, 23.38, 3.55. HRMS (ESI) m / z: calcd for C 18 H 22 N4O2SCl + [M+H] + : 393.1152, found: 393.1162.

[0059] Example 5: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3- (thiazol-4-ylmethyl)pyrimidine-2,4(lH,3H)-dione (4e).

[0060] Using intermediate 2 and 4-(bromomethyl)thiazole, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method is the same as that of compound 4a. The product 4d is collected as a white solid in a yield of 50% and a purity of 95.12%. 1H NMR (400 MHz, Chloroform-d6) δ 8.73 (s, 1H), 7.29 (s, 1H), 5.29 (s, 2H), 5.26 (s, 1H), 4.57 - 4.51 (m, 2H), 3.33 (d, J = 9.9 Hz, 1H), 3.21 (d, J = 12.0 Hz, 1H), 3.02 (tt, J = 8.7, 3.9 Hz, 1H), 2.70 (t, J = 10.9 Hz, 1H), 2.50 (t, J = 10.2 Hz, 1H), 2.02 - 1.93 (m, 1H), 1.90 - 1.83 (m, 1H), 1.81 (t, J = 2.0 Hz, 3H), 1.69 (s, 2H), 1.37 - 1.22 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 162.59, 159.46, 152.66, 152.60, 152.16, 116.31, 89.18, 80.20, 73.75, 59.31, 51.45, 47.53, 40.29, 35.57, 33.16, 23.24, 3.67. HRMS (ESI) m / z: calcd for C 17 H 22 N5O2S + [M+H] + : 360.1494, found: 360.1505.

[0061] Example 6: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3-((2- chlorothiazol-4-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4f).

[0062] Using intermediate 2 and 4-(bromomethyl)-2-chlorothiazole, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method is the same as that of compound 4a, the product 4f is collected as a yellow solid, yield: 51%, purity: 96.94%. 1H NMR (400 MHz, Chloroform-d6) δ 7.63 (s, 1H), 5.24 (s, 1H), 5.17 (s, 2H), 4.54 (d, J = 1.9 Hz, 2H), 3.34 (d, J = 9.8 Hz, 1H), 3.23 (d, J = 12.1 Hz, 1H), 3.04 (tt, J = 8.7, 3.9 Hz, 1H), 2.72 (t, J = 11.3 Hz, 1H), 2.52 (t, J = 10.5 Hz, 1H), 2.04 - 1.95 (m, 1H), 1.84 (t, J = 1.9 Hz, 3H), 1.75 - 1.63 (m, 1H), 1.58 (s, 2H), 1.38 - 1.24 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 162.11, 159.75, 152.56, 151.76, 141.90, 134.79, 88.77, 80.58, 73.42, 59.22, 51.38, 47.48, 36.39, 35.78, 33.13, 23.20, 3.66. HRMS (ESI) m / z: calcd for C 17 H 21 N5O2SCl + [M+H] + : 394.1104, found: 394.1105.

[0063] Example 7: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3-((2- methylthiazol-4-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4g).

[0064] Using intermediate 2 and 4-(bromomethyl)-2-methylthiazole, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method was the same as that of compound 4a, and the product 4g was collected as a yellow solid in a yield of 44% and a purity of 95.19%. 1H NMR (400 MHz, Chloroform-d6) δ 7.00 (s, 1H), 5.28 (s, 1H), 5.22 (s, 2H), 4.56 (d, J = 2.4 Hz, 2H), 3.36 (d, J = 11.6 Hz, 1H), 3.24 (d, J = 11.3 Hz, 1H), 3.10 - 3.02 (m, 1H), 2.75 (d, J = 11.1 Hz, 1H), 2.68 (s, 3H), 2.53 (d, J = 10.6 Hz, 1H), 2.05 - 1.97 (m, 1H), 1.94 - 1.86 (m, 1H), 1.83 (t, J = 2.3 Hz, 3H), 1.74 (d, J = 3.7 Hz, 2H), 1.39 - 1.27 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 165.48, 162.41, 159.39, 152.00, 151.24, 115.18, 88.92, 80.02, 73.77, 59.12, 51.34, 47.39, 40.47, 35.50, 33.04, 23.12, 19.00, 3.52. HRMS (ESI) m / z: calcd for C 18 H 24 N5O2S + [M+H] + : 374.1651, found: 374.1664.

[0065] Example 8: Preparation of (R)-methyl 5-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l- yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)furan-2-carboxylate (4h).

[0066] Using intermediate 2 and methyl 5-(bromomethyl)-2-furoate, (R)-3-aminopiperidine dihydrochloride as raw materials, the synthetic method was the same as that of compound 4a, and the product 4h was collected as a white solid in a yield of 61% and a purity of 95.96%. 1 H NMR (400 MHz, DMSO-d6) δ 7.23 (s, 1H), 6.45 (s, 1H), 5.20 (d, J = 4.0 Hz, 1H), 5.00 (s, 2H), 4.48 (s, 2H), 3.84 - 3.65 (m, 3H), 2.88 - 2.64 (m, 3H), 2.51 (s, 2H), 1.76 (d, J = 16.8 Hz, 5H), 1.56 (d, J = 12.8 Hz, 2H), 1.24 (dq, J = 38.0, 13.6, 12.4 Hz, 2H). 13C NMR (100 MHz, Chloroform-d6) d 162.19, 159.60, 159.05, 154.68, 151.88, 143.55, 119.04, 110.59, 88.90, 80.38, 73.57, 59.10, 51.78, 51.44, 47.47, 37.51, 35.70, 33.01, 23.19, 3.62. HRMS (ESI) m / z: calcd for C 20 H 25 N4O5[M+H] + = 401.1825, found [M+H] + = 401.1843.

[0067] Example 9: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3-((3,5- dimethylisoxazol-4-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4i).

[0068] Synthesis method same as compound 4a, collected product 4i as white solid, yield: 65%, purity: 95.65%. 1 H NMR (400 MHz, Chloroform-d6) d 5.17 (s, 1H), 4.82 (s, 2H), 4.50 (q, J = 2.5 Hz, 2H), 3.29 (d, J = 15.7 Hz, 1H), 3.18 (d, J = 11.7 Hz, 1H), 3.00 (tt, J = 8.7, 3.8 Hz, 1H), 2.68 (t, J = 11.1 Hz, 1H), 2.46 (s, 4H), 2.28 (s, 3H), 2.01 - 1.92 (m, 1H), 1.89 - 1.81 (m, 1H), 1.79 (t, J = 4.7 Hz, 3H), 1.73 - 1.61 (m, 1H), 1.49 (s, 2H), 1.34 - 1.18 (m, 1H). 13 C NMR (100 MHz, Chloroform-d6) d 168.75, 162.58, 160.11, 159.44, 152.01, 110.01, 88.93, 80.33, 73.65, 59.33, 51.45, 47.50, 35.51, 33.19, 33.01, 23.19, 11.56, 10.66, 3.62. HRMS (ESI) m / z: calcd for C 19 H 26 N5O3 + [M+H]+ : 372.2036, found: 372.2066.

[0069] Example 10: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3- ((l,3-dimethyl-lH-pyrazol-5-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4j).

[0070] The starting material was intermediate 2 and 5-(bromomethyl)-l,3-dimethyl-lH- pyrazole, (R)-3-aminopiperidine dihydrochloride, and the synthetic method was the same as compound 4a. The product 4j was collected as a white solid in a yield of 58% and a purity of 96.35%. 1 H NMR (400 MHz, Chloroform-d6) δ 6.06 (s, 1H), 5.14 (s, 1H), 4.97 (s, 2H), 4.52 - 4.38 (m, 2H), 3.87 (s, 3H), 3.25 (d, J = 13.3 Hz, 1H), 3.14 (d, J = 11.5 Hz, 1H), 2.98 - 2.90 (m, 1H), 2.62 (t, J = 10.9 Hz, 1H), 2.47 - 2.37 (m, 1H), 2.10 (s, 3H), 1.94 - 1.87 (m, 1H), 1.84 - 1.77 (m, 1H), 1.75 (t, J = 4.7 Hz, 3H), 1.66 - 1.56 (m, 1H), 1.49 (s, 2H), 1.28 - 1.12 (m, 1H). 13 C NMR (100 MHz, Chloroform-d6) δ 162.23, 159.47, 151.90, 147.06, 138.47, 106.93, 88.87, 80.33, 73.58, 59.24, 51.38, 47.46, 36.53, 35.59, 34.78, 33.13, 23.17, 13.33, 3.62. HRMS (ESI) m / z: calcd for C 19 H 27 N6O2 + [M+H] + : 371.2195, found: 371.2207.

[0071] Example 11: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3- ((5-methyl-l,3,4-oxadiazol-2-yl)methyl)pyrimidine-2,4(lH,3H)-dione (4k).

[0072] Using intermediate 2 and 2-(bromomethyl)-5-methyl-1,3,4-oxadiazole, (R)-3- aminopiperidine dihydrochloride as materials, with the same synthesis method of compound 4a, the product 4k was collected as a white solid in a yield of 62% and a purity of 96.50%. 1 H NMR (400 MHz, Chloroform-d6) δ 5.28 (s, 2H), 5.27 (s, 1H), 4.60 - 4.50 (m, 2H), 3.37 (d, J = 13.1 Hz, 1H), 3.26 (d, J = 11.6 Hz, 1H), 3.08 - 2.97 (m, 1H), 2.75 (t, J = 10.9 Hz, 1H), 2.55 (d, J = 10.5 Hz, 1H), 2.49 (s, 3H), 2.04 - 1.96 (m, 1H), 1.93 - 1.85 (m, 1H), 1.82 (t, J = 4.7 Hz, 3H), 1.75 - 1.65 (m, 1H), 1.54 (s, 1H), 1.38 - 1.19 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 163.94, 162.26, 161.92, 159.92, 151.83, 88.72, 80.58, 73.45, 59.24, 51.45, 47.53, 35.90, 35.52, 33.10, 23.24, 11.00, 3.67. HRMS (ESI) m / z: calcd for C 17 H 23 N6O3 + [M+H] + : 359.1832, found: 359.1854.

[0073] Example 12: Preparation of (R)-6-(3-aminopiperidin-1-yl)-1-(but-2-yn-1-yl)-3-(4- methylbenzyl)pyrimidine-2,4(1H,3H)-dione (4l).

[0074] Using intermediate 2 and 1-(bromomethyl)-4-methylbenzene, (R)-3-aminopiperidine dihydrochloride as materials, with the same synthesis method of compound 4a, the product 4l was collected as a white solid in a yield of 57% and a purity of 95.07%. 1H NMR (400 MHz, Chloroform-d6) δ 7.44 (d, J = 8.1 Hz, 2H), 7.12 (d, J = 7.8 Hz, 2H), 5.24 (s, 1H), 5.07 (s, 2H), 4.53 (q, J = 2.4 Hz, 2H), 3.32 (d, J = 11.8 Hz, 1H), 3.21 (d, J = 11.9 Hz, 1H), 3.07 - 2.99 (m, 1H), 2.70 (s, 1H), 2.49 (s, 1H), 2.32 (s, 3H), 1.99 (q, J = 5.6, 4.9 Hz, 1H), 1.91 - 1.86 (m, 1H), 1.83 (t, J = 2.3 Hz, 3H), 1.75 - 1.64 (m, 1H), 1.50 (s, 2H), 1.33 - 1.27 (m, 1H). 13 C NMR (100 MHz, Chloroform-d6) δ 162.88, 159.28, 152.21, 137.06, 134.15, 129.18, 129.18, 128.91, 128.91, 89.15, 80.10, 73.84, 59.24, 51.42, 47.49, 43.95, 35.46, 33.13, 23.23, 21.10, 3.62. HRMS (ESI) m / z: calcd for C 21 H 27 N4O2 + [M+H] + : 367.2134, found: 367.2148.

[0075] Example 13: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo- pyrimidin-l(2H)-yl)methyl)-N-(pyridin-3-ylmethyl)benzamide (9a).

[0076] To a 25 ml DMF solution, add intermediate 2 (10.00 g, 0.05 mol), sodium hydride (NaH) (2.40 g, 0.10 mol) and lithium bromide (LiBr) (13.00 g, 0.15 mol) under ice bath condition, stir for 20 min. Then add methyl 3-bromomethylbenzoate (13.70 g, 0.06 mol) to the reaction solution, and react at room temperature overnight. Post-treatment: add water to the reaction solution to terminate the reaction, extract with ethyl acetate, combine the organic phase, wash with water, then dry over anhydrous sodium sulfate for 2 h, and purify by silica gel chromatography column (PE:EA = 6:1) to obtain intermediate 4. To a 50 ml ethanol solution, add intermediate 4 (13.90 g, 0.04 mol), (R)-3-Boc-aminopiperidine (9.60 g, 0.05 mol), sodium bicarbonate (16.80 g, 0.20 mol) and 3.00 g activated molecular sieves (4A), and react at 100°C for 4 h. Post-treatment: concentrate the reaction and purify by silica gel chromatography column to obtain intermediate 5 (PE:EA = 3:1). Dissolve intermediate 5 (18.00 g, 0.04 mol) in 15 ml methanol, slowly add NaOH solution (1M) to the reaction solution while stirring, and react at room temperature for 10 h. Post-treatment: after complete hydrolysis, slowly add dilute hydrochloric acid (1M) dropwise to the reaction solution, and continuously precipitate white solid until the pH is adjusted to 3-4. Filter, and the obtained white solid is intermediate 6. To a 10 ml dichloromethane solution, add intermediate 6 (0.30 g, 0.60 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) (0.170 g, 0.90 mmol), 1-hydroxybenzotriazole (HOBT) (0.12 g, 0.90 mmol) and DIPEA (0.3 ml, 1.80 mmol), stir for 15 min, then add commercially available compound 7a, i.e. 3-aminomethylpyridine (0.60 mmol), and react at room temperature overnight. Post-treatment: concentrate the reaction and purify by silica gel chromatography column to obtain intermediate 8a. Dissolve intermediate 8a in 8 ml dichloromethane, slowly add trifluoroacetic acid (8 ml, TFA) dropwise under ice bath, and react at room temperature for 2-4 h. Post-treatment: add saturated NaHCO3 solution to the reaction solution to adjust the pH to 7-8 weak alkaline, extract the reaction solution with dichloromethane, combine the organic phase, wash with saturated brine 3-4 times, then dry over anhydrous sodium sulfate for 2 h, and concentrate the liquid to obtain the final product 9a. The collected product 9a is a white solid, with a yield of 29% and a purity of 96.14%. 1H NMR (400 MHz, Chloroform-d6) δ 8.60 (d, J = 1.7 Hz, 1H), 8.52 (dd, J = 4.7, 1.7 Hz, 1H), 7.86 (s, 1H), 7.78 (d, J = 7.8 Hz, 1H), 7.73 (d, J = 7.8 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.38 (t, J = 7.7 Hz, 1H), 7.27 (t, J = 4.8, 0.8 Hz, 1H), 7.03 (t, J = 5.3 Hz, 1H), 5.23 (s, 1H), 5.11 (s, 2H), 4.64 (d, J = 5.9 Hz, 2H), 4.50 (q, J = 2.4 Hz, 2H), 3.37 - 3.29 (m, 1H), 3.28 - 3.20 (m, 1H), 3.12 (s, 1H), 2.77 (s, 1H), 2.61 (s, 1H), 2.06 (s, 3H), 1.82 (t, J = 2.3 Hz, 3H), 1.73 - 1.64 (m, 1H), 1.40 - 1.28 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 167.44, 162.90, 159.50, 152.24, 149.25, 148.80, 137.46, 135.79, 134.20, 134.15, 132.77, 128.82, 127.25, 127.01, 123.60, 89.04, 80.41, 73.62, 58.98, 51.46, 47.50, 44.09, 41.44, 35.69, 32.91, 23.18, 3.61. HRMS (ESI) m / z: calcd for C 27 H 31 N6O3 + [M+H] + : 487.2458, found: 487.2467.

[0077] Example 14: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(thiophen-2-ylmethyl)benzamide (9b).

[0078] Using commercially available compound 7b, 2-aminomethylthiophene, as starting material, the synthesis method is the same as compound 9a. The product 9b is collected as yellow solid in 26% yield with a purity of 95.10%. 1H NMR (400 MHz, Chloroform-d6) δ 7.84 (s, 1H), 7.77 (d, J = 7.7 Hz, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.38 (t, J = 7.7 Hz, 1H), 7.24 (d, J = 5.1 Hz, 1H), 7.05 (d, J = 3.4 Hz, 1H), 6.97 (t, J = 4.3 Hz, 1H), 6.71 (t, J = 5.6 Hz, 1H), 5.25 (s, 1H), 5.12 (s, 2H), 4.81 (d, J = 5.6 Hz, 2H), 4.52 (s, 2H), 3.35 (d, J = 11.6 Hz, 1H), 3.24 (d, J = 11.8 Hz, 1H), 3.06 (dd, J = 8.9, 4.9 Hz, 1H), 2.79 - 2.68 (m, 1H), 2.56 (s, 1H), 2.08 (s, 2H), 1.93 - 1.86 (m, 1H), 1.82 (t, J = 1.8 Hz, 3H), 1.75 - 1.63 (m, 1H), 1.29 (d, J = 12.3 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 167.01, 162.90, 159.43, 152.22, 140.91, 137.46, 134.26, 132.68, 128.78, 127.23, 126.91, 126.89, 126.23, 125.28, 89.11, 80.41, 73.64, 51.49, 47.51, 44.09, 38.76, 35.70, 32.72, 29.71, 23.22, 3.68. HRMS (ESI) m / z: calcd for C 26 H 30 N5O3S + [M+H] + : 492.2069, found: 492.2088.

[0079] Example 15: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-isopropylbenzamide (9c).

[0080] Using commercially available compound 7c, isopropylamine, as the starting material, the synthesis method is the same as that of compound 9a. The product 9c is collected as a white solid in a yield of 19% and a purity of 96.61%. 1H NMR (400 MHz, Chloroform-d6) δ 7.80 (s, 1H), 7.72 (d, J = 6.2 Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.37 (t, J = 8.3 Hz, 1H), 6.05 (d, J = 8.0 Hz, 1H), 5.26 (s, 1H), 5.13 (s, 2H), 4.53 (t, J = 2.2 Hz, 2H), 4.33 - 4.23 (m, 1H), 3.41 - 3.33 (m, 1H), 3.25 (d, J = 11.3 Hz, 1H), 3.12 - 3.04 (m, 1H), 2.75 (s, 1H), 2.57 (s, 1H), 2.07 - 1.98 (m, 1H), 1.89 (d, J = 4.4 Hz, 1H), 1.83 (t, J = 2.3 Hz, 3H), 1.40 (d, J = 3.5 Hz, 1H), 1.32 - 1.25 (m, 9H). 13 C NMR (100 MHz, DMSO-d6) δ 165.77, 162.29, 159.92, 152.24, 137.97, 135.44, 130.43, 128.56, 127.30, 126.25, 87.98, 80.15, 75.01, 59.04, 51.24, 47.65, 43.80, 41.45, 36.05, 33.16, 23.37, 22.78, 22.78, 3.54. HRMS (ESI) m / z: calcd for C 24 H 32 N5O3 + [M+H] + : 438.2505, found: 438.2512.

[0081] Example 16: Preparation of (R)-6-(3-aminopiperidin-l-yl)-l-(but-2-yn-l-yl)-3-(3-(pyrrolidine- 1-carbonyl)benzyl)pyrimidine-2,4(lH,3H)-dione (9d).

[0082] Using commercially available compound 7d, tetrahydropyrrole, as starting material, the synthesis method is the same as compound 9a, the product 9d is collected as a white solid in 26% yield with a purity of 95.93%. 1H NMR (400 MHz, Chloroform-d6) δ 7.52 (d, J = 7.3 Hz, 2H), 7.38 - 7.30 (m, 2H), 5.23 (s, 1H), 5.08 (s, 2H), 4.50 (q, J = 2.4 Hz, 2H), 3.57 (t, J = 6.9 Hz, 2H), 3.48 (s, 4H), 3.44 - 3.37 (m, 2H), 3.30 (s, 2H), 2.91 (s, 2H), 2.04 (s, 1H), 1.94 (q, J = 6.5 Hz, 2H), 1.86 (t, J = 6.6 Hz, 2H), 1.80 (s, 3H), 1.68 (s, 1H), 1.40 - 1.21 (m, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 168.43, 162.25, 159.52, 152.07, 137.78, 137.59, 129.58, 128.66, 126.90, 126.33, 88.85, 80.21, 74.95, 53.02, 51.80, 49.38, 46.53, 46.38, 43.82, 36.03, 36.01, 27.84, 26.42, 24.35, 3.55. HRMS (ESI) m / z: calcd for C 25 H 32 N5O3 + [M+H] + : 450.2505, found: 450.2504.

[0083] Example 17: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)- 2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-cyclohexylbenzamide (9e).

[0084] Using commercially available compound 7e, cyclohexylamine, as the starting material, the synthesis method was the same as that of compound 9a. The product 9e was collected as a white solid in a yield of 32% and a purity of 96.22%. 1H NMR (400 MHz, Chloroform-d6) δ 7.80 (s, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 6.14 (d, J = 8.1 Hz, 1H), 5.26 (s, 1H), 5.12 (s, 2H), 4.52 (q, J = 2.4 Hz, 2H), 4.01 - 3.91 (m, 1H), 3.34 (d, J = 11.5 Hz, 1H), 3.23 (d, J = 12.1 Hz, 1H), 3.11 - 3.02 (m, 1H), 2.74 (t, J = 10.9 Hz, 1H), 2.57 (d, J = 10.3 Hz, 1H), 2.45 (s, 3H), 2.03 (d, J = 6.4 Hz, 2H), 1.89 (dt, J = 13.6, 4.1 Hz, 1H), 1.83 (t, J = 4.6 Hz, 3H), 1.79 - 1.74 (m, 2H), 1.72 - 1.63 (m, 2H), 1.44 - 1.36 (m, 2H), 1.31 - 1.19 (m, 4H). 13 C NMR (100 MHz, DMSO-d6) δ 165.80, 162.30, 159.91, 152.24, 137.95, 135.49, 130.41, 128.56, 127.34, 126.31, 88.00, 80.15, 79.63, 75.00, 58.87, 51.25, 48.82, 47.61, 43.80, 36.04, 32.86 (2C), 30.14, 25.73, 25.41 (2C), 3.54. HRMS (ESI) m / z: calcd for C 27 H 36 N5O3 + [M+H] + : 478.2818, found: 478.2831.

[0085] Example 18: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(cyclohexylmethyl)benzamide (9f).

[0086] Using commercially available compound 7f, cyclohexylmethylamine, as the starting material, the synthesis method was the same as that of compound 9a. The product 9f was collected as a white solid in a yield of 30% and a purity of 97.59%. 1H NMR (400 MHz, Chloroform-d6) δ 7.82 (s, 1H), 7.73 (d, J = 7.9 Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.37 (t, J = 7.7 Hz, 1H), 6.40 (t, J = 6.0 Hz, 1H), 5.25 (s, 1H), 5.12 (s, 2H), 4.52 (q, J = 2.4 Hz, 2H), 3.34 (d, J = 11.4 Hz, 1H), 3.28 (t, J = 6.5 Hz, 2H), 3.23 (d, J = 11.8 Hz, 1H), 3.07 - 2.98 (m, 1H), 2.72 (t, J = 10.4 Hz, 1H), 2.52 (t, J = 10.3 Hz, 1H), 2.04 - 1.95 (m, 1H), 1.92 - 1.85 (m, 1H), 1.83 (t, J = 2.3 Hz, 3H), 1.75 (d, J = 11.6 Hz, 7H), 1.70 - 1.65 (m, 2H), 1.62 - 1.54 (m, 1H), 1.29 - 1.21 (m, 3H), 1.06 - 0.93 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 167.47, 162.88, 159.46, 152.26, 137.37, 135.06, 132.23, 128.69, 127.09, 126.72, 89.12, 80.35, 73.68, 59.27, 51.46, 47.54, 46.27, 44.10, 38.03, 35.65, 33.11, 30.95 (2C) 26.42, 25.84 (2C), 23.23, 3.64. HRMS (ESI) m / z: calcd for C 28 H 38 N5O3 + [M+H] + : 492.2975, found: 492.2970.

[0087] Example 19: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)- 2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(2-cyclohexylethyl)benzamide (9g).

[0088] Using commercially available compound 7g, 2-cyclohexylethylamine, as the starting material, the synthesis method is the same as that of compound 9a. The product 9g is collected as a white solid in a yield of 45% and a purity of 95.53%. 1H NMR (400 MHz, Chloroform-d6) δ 7.81 (s, 1H), 7.74 (d, J = 7.8 Hz, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.37 (t, J = 7.7 Hz, 1H), 6.25 (s, 1H), 5.26 (s, 1H), 5.13 (s, 2H), 4.53 (t, J = 2.4 Hz, 2H), 3.50 - 3.43 (m, 2H), 3.35 (d, J = 11.5 Hz, 1H), 3.23 (d, J = 11.7 Hz, 1H), 3.06 (s, 1H), 2.74 (s, 1H), 2.54 (s, 1H), 2.21 (s, 2H), 2.01 (d, J = 22.3 Hz, 2H), 1.83 (t, J = 2.5 Hz, 3H), 1.77 - 1.68 (m, 4H), 1.52 (q, J = 7.2 Hz, 2H), 1.36 - 1.14 (m, 7H), 0.98 (td, J = 12.0, 2.9 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 167.35, 162.91, 159.44, 152.23, 137.33, 135.00, 132.34, 128.73, 127.08, 126.79, 89.15, 80.40, 73.65, 59.03, 51.48, 47.46, 44.11, 37.97, 37.12, 35.65, 35.49, 33.20 (2C), 32.90, 26.53, 26.23 (2C), 23.19, 3.68. HRMS (ESI) m / z: calcd for C 29 H 40 N5O3 + [M+H] + : 506.3131, found: 506.3145.

[0089] Example 20: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(3-fluorophenethyl)benzamide (9h).

[0090] Using commercially available compound 7h, 2-(3-fluorophenyl)ethan-l-amine as starting material, the synthesis method is the same as compound 9a, the product 9h is collected as a white solid in 39% yield, 95.04% purity. 1H NMR (400 MHz, Chloroform-d6) δ 7.79 (s, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.66 (d, J = 7.8 Hz, 1H), 7.38 (t, J = 7.7 Hz, 1H), 7.34 - 7.29 (m, 1H), 7.04 (d, J = 7.7 Hz, 1H), 6.99 - 6.92 (m, 2H), 6.38 (t, J = 5.6 Hz, 1H), 5.25 (s, 1H), 5.12 (s, 2H), 4.53 (q, J = 2.4 Hz, 2H), 3.70 (q, J = 7.0 Hz, 2H), 3.35 (d, J = 11.6 Hz, 1H), 3.24 (d, J = 11.1 Hz, 1H), 3.10 - 3.01 (m, 1H), 2.95 (t, J = 7.1 Hz, 2H), 2.75 (d, J = 11.5 Hz, 1H), 2.53 (s, 1H), 2.04 - 1.97 (m, 1H), 1.89 (s, 2H), 1.83 (t, J = 2.3 Hz, 3H), 1.75 - 1.64 (m, 1H), 1.35 - 1.29 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 166.50, 162.26, 159.66, 157.71, 152.14, 137.96, 135.29, 130.65, 130.50, 128.63, 128.04, 127.72, 127.14, 126.15, 120.74, 111.13, 88.52, 79.66, 74.97, 55.72, 55.34, 51.60, 46.96, 43.88, 36.01, 30.28, 29.87, 22.37, 3.55. HRMS (ESI) m / z: calcd for C 29 H 33 N5O3F + [M+H] + : 518.2567, found: 518.2582.

[0091] Example 21: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(3-chlorophenethyl)benzamide (9i).

[0092] Using commercially available compound 7i, 2-(3-chlorophenyl)ethan-l-amine as the starting material, the synthesis method is the same as compound 9a, and the product 9i is collected as a white solid in a yield of 32% and a purity of 96.88%. 1H NMR (400 MHz, Chloroform-d6) δ 7.79 (s, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.64 (d, J = 6.9 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 7.28 (s, 1H), 7.25 - 7.20 (m, 2H), 7.14 (d, J = 7.4 Hz, 1H), 6.53 (t, J = 5.9 Hz, 1H), 5.24 (s, 1H), 5.10 (s, 2H), 4.51 (q, J = 2.5 Hz, 2H), 3.67 (q, J = 6.8 Hz, 2H), 3.34 (d, J = 10.2 Hz, 1H), 3.23 (d, J = 10.8 Hz, 1H), 3.11 - 3.02 (m, 1H), 2.91 (t, J = 7.2 Hz, 2H), 2.75 (d, J = 11.0 Hz, 1H), 2.61 - 2.50 (m, 1H), 1.99 (d, J = 8.9 Hz, 2H), 1.93 - 1.85 (m, 1H), 1.82 (t, J = 2.3 Hz, 3H), 1.75 - 1.61 (m, 1H), 1.39 - 1.27 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 166.65, 162.27, 159.91, 152.24, 142.64, 138.09, 135.18, 133.37, 130.71, 130.59, 129.02, 128.67, 127.96, 127.16, 126.56, 126.11, 88.00, 80.15, 75.02, 58.99, 51.25, 47.64, 43.80, 36.05, 35.01, 33.11, 23.34, 21.86, 3.55. HRMS (ESI) m / z: calcd for C 29 H 33 N5O3Cl + [M+H] + : 534.2272, found: 534.2281.

[0093] Example 22: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(3-bromophenethyl)benzamide (9j).

[0094] Using commercially available compound 7j, 2-(3-bromophenyl)ethan-l-amine as starting material, the synthesis method is the same as compound 9a, the product 9j is collected as a yellow solid in 32% yield, 95.04% purity. 1H NMR (400 MHz, DMSO-d6) δ 8.57 (t, J = 5.5 Hz, 1H), 7.73 (s, 1H), 7.66 (d, J = 5.9 Hz, 1H), 7.46 (s, 1H), 7.39 (d, J = 6.3 Hz, 3H), 7.25 (d, J = 6.4 Hz, 2H), 5.21 (s, 1H), 4.98 (s, 2H), 4.49 (q, J = 2.6 Hz, 2H), 3.48 (q, J = 6.7 Hz, 2H), 3.23 (dd, J = 11.7, 3.5 Hz, 1H), 3.16 (d, J = 11.9 Hz, 1H), 2.85 (t, J = 7.1 Hz, 3H), 2.71 (t, J = 11.0 Hz, 1H), 2.45 (d, J = 10.2 Hz, 1H), 1.84 (d, J = 9.0 Hz, 2H), 1.77 (t, J = 2.4 Hz, 3H), 1.75 (s, 1H), 1.63 - 1.51 (m, 1H), 1.21 (d, J = 12.6 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 166.68, 162.28, 159.89, 152.24, 142.93, 138.08, 135.19, 131.91, 130.90, 130.73, 129.44, 128.67, 128.33, 127.18, 126.13, 122.09, 88.04, 80.16, 75.02, 58.74, 51.28, 47.57, 43.82, 40.99, 36.05, 34.98, 32.89, 23.29, 3.55. HRMS (ESI) m / z: calcd for C 29 H 33 N5O3Br + [M+H] + : 578.1767, found: 578.1774.

[0095] Example 23: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)- 2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(3-methylphenethyl)benzamide (9k).

[0096] Using commercially available compound 7k, 2-(3-methylphenyl)ethan-l-amine as the starting material, the synthetic method is the same as compound 9a, the product 9k is collected as a white solid in a yield of 52% with a purity of 95.14%. 1H NMR (400 MHz, Chloroform-d6) δ 7.79 (s, 1H), 7.68 (d, J = 7.7 Hz, 1H), 7.64 (d, J = 7.8 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 7.23 (t, J = 7.4 Hz, 1H), 7.10 - 7.06 (m, 2H), 7.05 (d, J = 1.9 Hz, 1H), 6.49 (t, J = 5.7 Hz, 1H), 5.25 (s, 1H), 5.11 (s, 2H), 4.52 (q, J = 2.4 Hz, 2H), 3.68 (q, J = 6.8 Hz, 2H), 3.34 (d, J = 8.7 Hz, 1H), 3.22 (d, J = 11.4 Hz, 1H), 3.09 - 3.01 (m, 1H), 2.90 (t, J = 7.2 Hz, 2H), 2.79 - 2.67 (m, 1H), 2.60 - 2.48 (m, 1H), 2.36 (s, 3H), 2.01 (d, J = 12.3 Hz, 2H), 1.92 - 1.85 (m, 1H), 1.83 (t, J = 2.4 Hz, 3H), 1.75 - 1.64 (m, 1H), 1.33 - 1.27 (m, 2H). 13 C NMR (100 MHz, Chloroform-d6) δ 167.45, 162.90, 159.47, 152.23, 138.95, 138.28, 137.35, 134.86, 132.35, 129.64, 128.68, 128.55, 127.24, 126.65, 125.84, 89.17, 80.40, 73.66, 59.02, 51.50, 47.45, 44.11, 41.33, 35.70, 32.95, 31.50, 29.68, 23.15, 21.36, 3.61. HRMS (ESI) m / z: calcd for C 30 H 36 N5O3 + [M+H] + : 514.2818, found: 514.2812.

[0097] Example 24: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(2-fluorophenethyl)benzamide (9l).

[0098] Using commercially available compound 7l, 2-(2-fluorophenyl)ethan-l-amine as starting material, the synthesis method is the same as compound 9a, the product 9l is collected as a white solid in 52% yield, 95.08% purity.1 H NMR (400 MHz, Chloroform-d6) δ 7.78 (s, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 7.6 Hz, 1H), 7.34 (t, J = 7.7 Hz, 1H), 7.23 (ddd, J = 14.0, 7.9, 5.7 Hz, 2H), 7.12 - 7.00 (m, 2H), 6.66 (t, J = 5.8 Hz, 1H), 5.24 (s, 1H), 5.09 (s, 2H), 4.55 - 4.48 (m, 2H), 3.67 (q, J = 6.6 Hz, 2H), 3.33 (d, J = 11.5 Hz, 1H), 3.23 (d, J = 12.1 Hz, 1H), 3.11 - 3.02 (m, 1H), 2.97 (t, J = 7.1 Hz, 2H), 2.70 (s, 2H), 2.56 (t, J = 10.3 Hz, 1H), 2.02 - 1.96 (m, 1H), 1.92 - 1.85 (m, 1H), 1.81 (t, J = 4.7 Hz, 3H), 1.74 - 1.62 (m, 1H), 1.36 - 1.27 (m, 2H). 13 CNMR (100 MHz, DMSO-d6) δ 166.63, 162.29, 159.90, 152.24, 138.07, 135.17, 131.69, 130.71, 128.67, 127.11, 126.61, 126.45, 126.12, 124.84, 115.68, 115.47, 88.02, 80.16, 79.63, 75.00, 58.80, 51.27, 47.59, 43.81, 36.05, 32.95, 29.02, 23.30, 3.54. HRMS (ESI) m / z: calcd for C 29 H 33 N5O3F + [M+H] + : 518.2567, found: 518.2570.

[0099] Example 25: Preparation of (R)-3-((4-(3-aminopiperidin-l-yl)-3-(but-2-yn-l-yl)-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)-N-(2-chlorophenethyl)benzamide (9m).

[0100] Using commercially available compound 7m, 2-(2-chlorophenyl)ethan-l-amine as starting material, the same synthetic procedure as compound 9a, product 9m was collected as white solid in 26% yield, 95.87% purity. 1H NMR (400 MHz, Chloroform-d6) δ 7.79 (s, 1H), 7.71 (d, J = 7.7 Hz, 1H), 7.64 (d, J = 7.6 Hz, 1H), 7.40 - 7.34 (m, 2H), 7.29 - 7.27 (m, 1H), 7.27 - 7.17 (m, 2H), 6.51 (t, J = 5.9 Hz, 1H), 5.25 (s, 1H), 5.11 (s, 2H), 4.52 (d, J = 2.8 Hz, 2H), 3.71 (q, J = 6.7 Hz, 2H), 3.34 (d, J = 11.4 Hz, 1H), 3.23 (d, J = 11.9 Hz, 1H), 3.08 (q, J = 9.4, 8.2 Hz, 3H), 2.74 (s, 1H), 2.50 (s, 3H), 2.02 (s, 1H), 1.82 (t, J = 2.6 Hz, 3H), 1.74 - 1.65 (m, 1H), 1.37 - 1.26 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 166.67, 162.30, 159.91, 152.23, 138.06, 137.34, 135.15, 133.60, 131.61, 130.72, 129.70, 128.67, 128.65, 127.72, 127.11, 126.15, 88.00, 80.17, 75.00, 58.82, 51.26, 47.59, 43.81, 39.31, 36.06, 33.29, 32.96, 23.31, 3.54. HRMS (ESI) m / z: calcd for C 29 H 33 N5O3Cl + [M+H] + : 534.2272, found: 534.2270.

[0101] Example 26: Preparation of (R)-6-(3-aminopyrrolidin-1-yl)-1-(but-2-yn-1-yl)-3-((2-oxo-1,2-dihydroquinolin-4-yl)methyl)pyrimidine-2,4(1H,3H)-dione (10a).

[0102] To a solution of 15 ml DMF was added intermediate 2 (5.00 g, 0.03 mol), sodium hydride (NaH) (1.20 g, 0.05 mol) and lithium bromide (LiBr) (6.50 g, 0.075 mol) were added under ice bath condition and stirred for 15 min. Then, commercially available compound 4-(bromomethyl)quinolin-2(lH)-one and 2-(bromomethyl)benzothiazole (0.030 mol) were added to the reaction solution, respectively, and reacted at room temperature overnight. Work-up: the reaction was terminated by adding water and extracted with ethyl acetate, the organic phases were combined, washed with water and dried over anhydrous sodium sulfate for 2 h, and then purified by silica gel chromatography column to obtain intermediates 3r and 3p.

[0103] To a solution of 10 ml ethanol was added intermediate 3r (2.50 mmol), (R)-l-bromopyrrolidin-3-amine (3.750 mmol), sodium bicarbonate (1.05 g, 12.50 mmol) and 100.00 mg activated molecular sieves (4A), respectively, and reacted at 100 °C for 2.5 h. Work-up: the reaction was concentrated and purified by silica gel chromatography column to obtain the final product 10a. The product 10a was collected as a white solid with a yield of 43% and a purity of 95.11%. 1 H NMR (400 MHz, DMSO-d6) δ 11.50 (s, 1H), 7.89 (d, J = 8.1 Hz, 1H), 7.55 (t, J = 7.7 Hz, 1H), 7.36 (d, J = 8.3 Hz, 1H), 7.24 (t, J = 7.7 Hz, 1H), 5.91 (s, 1H), 5.17 (s, 2H), 5.02 (s, 1H), 4.55 (s, 2H), 3.65 - 3.49 (m, 4H), 3.10 (t, J = 6.0 Hz, 1H), 2.07 - 1.96 (m, 1H), 1.81 (s, 3H), 1.75 - 1.63 (m, 1H), 1.40 - 1.15 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 161.85, 161.47, 156.93, 152.72, 146.70, 139.21, 131.08, 124.25, 122.39, 117.94, 116.79, 116.16, 80.72, 80.38, 75.34, 59.50, 51.10, 49.79, 40.99, 38.28, 34.17, 3.62. HRMS (ESI) m / z: calcd for C 22 H 24 N5O3 + [M+H] + ; 406.1879, found: 406.1883.

[0104] Example 27: Preparation of (R)-1-(but-2-yn-1-yl)-6-(3-hydroxypyrrolidin-1-yl)-3-((2-oxo-1,2- dihydroquinolin-4-yl)methyl)pyrimidine-2,4(1H,3H)-dione (10b).

[0105] Using intermediate 3r and (R)-1-bromopyrrolidin-3-ol as raw materials, the synthetic method is the same as that of compound 10a, and the product 10b is collected as a white solid in a yield of 58% and a purity of 95.00%. 1 H NMR (400 MHz, Chloroform-d6) δ 11.65 (s, 1H), 7.79 (d, J = 9.5 Hz, 1H), 7.49 (t, J = 8.3 Hz, 1H), 7.35 (d, J = 9.4 Hz, 1H), 7.28 - 7.23 (m, 1H), 6.33 (s, 1H), 5.41 (s, 2H), 5.20 (s, 1H), 4.60 (dt, J = 16.0, 2.3 Hz, 2H), 3.75 (td, J = 9.4, 7.1 Hz, 1H), 3.59 (dd, J = 10.9, 4.3 Hz, 1H), 3.50 - 3.41 (m, 2H), 2.98 (s, 1H), 2.14 - 2.10 (m, 1H), 1.82 (t, J = 2.3 Hz, 3H), 1.40 - 1.24 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 161.83, 161.48, 156.96, 152.70, 146.66, 139.24, 131.06, 124.25, 122.37, 117.96, 116.86, 116.17, 80.96, 80.42, 75.31, 69.34, 59.60, 49.27, 49.07, 38.27, 33.73, 3.59. HRMS (ESI) m / z: calcd for C 22 H 23 N4O4 + [M+H] + : 407.1719, found: 407.1716.

[0106] Example 28: Preparation of (R)-1-(but-2-yn-1-yl)-6-(3-hydroxypiperidin-1-yl)-3-((2-oxo-1,2- dihydroquinolin-4-yl)methyl)pyrimidine-2,4(1H,3H)-dione (10c).

[0107] Using intermediate 3r and (R)-1-bromopyrrolidin-3-ol as raw materials, the synthetic method is the same as that of compound 10a, and the product 10b is collected as a white solid in a yield of 58% and a purity of 95.00%.1 H NMR (400 MHz, Chloroform-d6) δ 12.15 (s, 1H), 7.77 (d, J = 8.2 Hz, 1H), 7.49 (t, J = 7.7 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.27 - 7.21 (m, 1H), 6.31 (s, 1H), 5.40 (s, 2H), 5.35 (s, 1H), 4.62 (p, J = 2.5 Hz, 2H), 4.02 - 3.91 (m, 1H), 3.31 - 3.21 (m, 1H), 3.14 (d, J = 11.8 Hz, 1H), 3.06 - 2.95 (m, 2H), 1.96 (t, J = 10.1 Hz, 1H), 1.82 (t, J = 2.3 Hz, 3H), 1.68 (q, J = 10.2, 9.7 Hz, 2H), 1.39 - 1.26 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 162.08, 161.82, 160.20, 152.15, 146.26, 139.21, 131.11, 124.24, 122.40, 117.88, 116.73, 116.17, 87.73, 80.22, 75.01, 65.11, 57.61, 51.04, 41.20, 36.37, 32.63, 22.35, 3.57. HRMS (ESI) m / z: calcd for C 23 H 25 N4O4 + [M+H] + : 421.1876, found: 421.1879.

[0108] Example 29: Preparation of (R)-6-(3-aminoazepan-l-yl)-3-(benzo[d]thiazol-2- ylmethyl)-l-(but-2-yn-l-yl)pyrimidine-2,4(lH,3H)-dione (10d).

[0109] Using intermediate 3p and (R)-l-bromoazepan-3-amine as the raw material, the synthetic method is the same as that of compound 10a, the product 10d is collected as a white solid in a yield of 32% and a purity of 95.21%. 1H NMR (400 MHz, Chloroform-d6) δ 8.02 (d, J = 8.1 Hz, 1H), 7.83 (d, J = 7.6 Hz, 1H), 7.45 (ddd, J = 8.3, 7.2, 1.3 Hz, 1H), 7.36 (ddd, J = 8.3, 7.2, 1.3 Hz, 1H), 5.55 (d, J = 1.5 Hz, 2H), 5.39 (s, 1H), 4.62 (qq, J = 17.0, 2.3 Hz, 2H), 3.43 (dd, J = 13.7, 3.7 Hz, 1H), 3.27 - 3.21 (m, 2H), 3.00 (dd, J = 13.7, 8.2 Hz, 1H), 2.06 - 1.97 (m, 1H), 1.96 - 1.87 (m, 1H), 1.83 (t, J = 2.4 Hz, 3H), 1.62 (t, J = 4.3 Hz, 3H), 1.56 - 1.49 (m, 1H), 1.39 (s, 1H), 1.30 (s, 1H), 1.27 (s, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 167.91, 161.74, 161.37, 152.72, 152.31, 135.07, 126.68, 125.68, 122.95, 122.68, 87.81, 80.09, 75.00, 61.25, 54.52, 51.36, 42.79, 37.12, 36.68, 27.93, 22.86, 3.56. HRMS (ESI) m / z: calcd for C 22 H 26 N5O2S + [M+H] + : 424.1807, found: 424.1812.

[0110] Example 30: Preparation of (R)-6-(3-aminopyrrolidin-l-yl)-3-(benzo[d]thiazol-2- ylmethyl)-l-(but-2-yn-l-yl)pyrimidine-2,4(lH,3H)-dione (10e).

[0111] Using intermediate 3p and (R)-l-bromopyrrolidine-3-amine as raw materials, the synthetic method is the same as that of compound 10a, and the product 10e is collected as a white solid in a yield of 40% and a purity of 95.40%. 1H NMR (400 MHz, Chloroform-d6) δ 8.01 (d, J = 8.8 Hz, 1H), 7.81 (d, J = 9.4 Hz, 1H), 7.44 (ddd, J = 8.3, 7.2, 1.3 Hz, 1H), 7.34 (ddd, J = 7.7, 7.3, 1.3 Hz, 1H), 5.54 (s, 2H), 5.16 (s, 1H), 4.57 (q, J = 2.1 Hz, 2H), 3.76 - 3.69 (m, 1H), 3.63 (dt, J = 9.9, 7.1 Hz, 1H), 3.55 (dd, J = 10.0, 5.6 Hz, 1H), 3.47 - 3.38 (m, 1H), 3.13 (dd, J = 10.0, 4.5 Hz, 1H), 2.18 (dq, J = 13.0, 6.7 Hz, 1H), 1.82 (t, J = 2.3 Hz, 3H), 1.77 (s, 2H), 1.38 - 1.27 (m, 1H). 13 C NMR (100 MHz, Chloroform-d6) δ 166.93, 161.91, 156.59, 152.89, 152.50, 135.39, 125.84, 124.99, 123.23, 121.46, 82.77, 80.64, 73.62, 59.53, 50.85, 49.74, 42.72, 37.78, 34.44, 3.69. HRMS (ESI) m / z: calcd for C 20 H 22 N5O2S + [M+H] + : 396.1494, found: 396.1505.

[0112] Example 31: Preparation of (R)-3-(benzo[d]thiazol-2-ylmethyl)-l-(but-2-yn-l-yl)-6-(3- hydroxypiperidin-l-yl)pyrimidine-2,4(lH,3H)-dione (10f).

[0113] Using intermediate 3p and (R)-l-bromopiperidin-3-ol as raw materials, the synthetic method was the same as that of compound 10a. The product 10f was collected as a white solid in a yield of 58% and a purity of 98.87%. 1H NMR(400MHz,Chloroform-d6)δ8.01(d,J=8.2Hz,1H),7.82(d,J=8.4Hz,1H),7.44(ddd,J=8.3, 7.2,1.3Hz,1H),7.35(ddd,J=8.2,7.2,1.2Hz,1H),5.54(s,2H),5.33(s,1H),4.62(qq,J=16.9 ,2.3Hz,2H),4.01–3.94(m,1H),3.25–3.18(m,1H),3.05(d,J=8.0Hz,1H),3.00(d,J=7.1Hz,1H ),2.51(s,1H),1.98–1.90(m,2H),1.82(t,J=2.3Hz,3H),1.73–1.62(m,2H),1.40–1.22(m,1H). 13 C NMR(100MHz,Chloroform-d6)δ166.36,162.27,159.85,152.82,152.03,135.35,125.93,125.11,123.25, 121.48,89.39,80.69,73.65,65.72,57.44,51.77,42.77,35.68,31.78,21.61,3.62.HRMS(ESI)m / z:calcd for C 21 H 23 N4O3S + [M+H] + :411.1491,found:411.1499.

[0114] Example 32: The fluorescence reporter strain method was used to determine the lasB inhibition rate of the derivatives against Pseudomonas aeruginosa PAO1.

[0115] Experimental method: The reporter strain was cultured in LB medium overnight and diluted with ABTGC medium to OD 600 The concentration of 0.02 was then added to a 96-well plate, with each well containing 75 μL of culture medium and 75 μL of bacterial solution containing the compound. A DMSO group (0.1% DMSO) and a blank group (culture medium only) were also set up. The plate was then incubated at 37°C in a microplate reader, and GFP fluorescence was measured every 15 minutes (excitation wavelength 485 nm, emission wavelength 535 nm) for at least 12 hours.

[0116] Table 2 Inhibition rate of uracil derivatives against Pseudomonas aeruginosa PAO1-lasB-gfp fluorescent reporter strain

[0117]

[0118] a Data are shown as mean ± SD. All experiments were performed in triplicate.

[0119] Experimental results: As shown in Table 2, all uracil derivatives showed good lasB inhibitory activity, and the inhibitory rates of compounds 4d, 4h, 9b, 9c, 9d, 9f, 9k, 9m, 10c, 10d, 10e, 10f on lasB all reached more than 50%, among which the lasB inhibitory rate of compound 10e was the highest, reaching 54.74%, which was the most active compound, and had the potential to be used as an anti-virulence compound to treat P. aeruginosa infection.

[0120] Example 48: Study on the antibacterial synergistic effect of compound 10e on ciprofloxacin (CIP) and tobramycin (Tob) in a mouse wound infection model

[0121] Experimental method: This animal experiment was carried out in accordance with the relevant national regulations on animal experiments. P. aeruginosa PAOl strain was used in this animal experiment, and female mice (5 weeks old, Babl / c) were purchased from SPF (Beijing) Biotechnology.co.Ltd. The mice were provided with sufficient food and water during the entire experimental process, and the living environment was maintained at 26°C with a 12h light / dark cycle. First, the mice were anesthetized with 4% chloral hydrate on day 0, and a 4-5mm circular wound was established on the back of each shaved mouse, and 5x10 8 CFU of PAOl was inoculated on the wound surface. After 24h of post-wound infection, different groups of mice were treated with different drugs (normal saline, 0.002mg / ml CIP, 0.002mg / ml CIP+2.5μM 10e, 0.0025mg / ml Tob, 0.0025mg / ml Tob+2.5μM 10e), respectively. After 3 days of continuous drug administration, the infected skin of the mouse wound was homogenized and CFU counting analysis was performed. After 9 days of continuous observation of the wound surface, the heart, liver, spleen, lung, and kidney of the mouse were taken out for H&E staining.

[0122] Experimental results: The results showed that Figure 1 and 2)Compared with the untreated control or the wound treated with 2.5 μM 10e, the amount of bacteria in the wound was greatly reduced after the combination of 0.002 mg / ml CIP, 0.0025 mg / ml Tob and 2.5 μM 10e, and was better than the single antibiotic administration group. At the same time, the monitoring of the wound area of mice also obtained similar results, the healing rate of the wound of mice in the combination administration group was accelerated, which proved the better antibacterial synergistic effect of 10e on CIP and Tob. In addition, the H&E staining results of lung tissue sections of mice showed that the combination of 10e and CIP or Tob had no obvious toxicity to mice.

[0123] The above description of the embodiments is only used to help understand the method of the present application and its core idea. It should be noted that for those skilled in the art of the art, without departing from the principles of the present application, the present application can be improved and modified, and these improvements and modifications also fall within the scope of the claims of the present application.

Claims

1. Use of a compound or a pharmaceutically acceptable salt thereof in the manufacture of a lasB inhibitor, the compound having the chemical structure shown below:

2. Use of a compound or a pharmaceutically acceptable salt thereof in the manufacture of an antibacterial agent, the compound having the chemical structure shown below:

3. The use according to claim 2, wherein the bacteria is a bacterium.

4. The use according to claim 2, wherein the bacteria is Pseudomonas aeruginosa.

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