A bergamot alcohol derivative and its use

Abstract

The application discloses a bergamot alcohol derivative or a medicinal salt thereof, and a structure general formula is as shown in the following formula: the bergamot alcohol derivative provided by the application is obtained by optimizing the structure of bergamot alcohol, and through antibacterial activity tests on common gram-positive bacteria such as drug-resistant staphylococcus aureus and gram-negative bacteria such as escherichia coli and klebsiella pneumoniae, it is found that the compound has excellent antibacterial activity on drug-resistant bacteria, and has good development value in preparation of anti-infection treatment drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry technology, specifically, it relates to a bergamot derivative and its applications. Background Technology

[0002] Antibiotic resistance is one of the major health problems facing society today. Excessive or inappropriate use of quinolone antibiotics can increase bacterial resistance, with methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRBBA), and drug-resistant Klebsiella pneumoniae becoming major types of nosocomial infections (Li Ping et al., Journal of Rational Clinical Drug Use, 2023, 4, 178-181; Niu Yinghui et al., Chinese Scientific and Technological Periodicals Database (Full Text Edition) Medicine and Health, 2022, 7, 262-264; Xing Lingyi et al., Journal of Tianjin University of Traditional Chinese Medicine, 2023, 1, 127-136). To address this global problem of antibiotic resistance, developing novel antibiotics with different mechanisms of action than quinolone antibiotics has become one effective solution.

[0003] Traditional Chinese medicine (TCM) has a long history of disease prevention and treatment, with a wide range of sources and few side effects, making it a potential source for developing novel antibacterial drugs. Psoralea corylifolia is an important medicinal plant with antibacterial, antitumor, antioxidant, anti-inflammatory, and immunomodulatory activities (Chen Ying et al., Practical Drugs and Clinical Applications, 2016, 9, 1184-1188). Bergerol, a derivative of psoralen, is one of the chemical components of Psoralea corylifolia and possesses anti-inflammatory, antitumor, and antioxidant effects (Wang Tianxiao et al., Chinese Journal of Traditional Chinese Medicine, 2013, 14, 2328-2333; Zhou Lishan et al., Chinese Journal of Traditional Chinese Medicine, 2019, 10, 2118-2123; Guo Jiangning et al., Chinese Medicinal Herbs, 2003, 3, 185-187). However, bergerol has relatively weak biological activity, thus requiring structural optimization to enhance its antibacterial and antitumor activities. Summary of the Invention

[0004] The purpose of this invention is to provide a bergamot derivative.

[0005] Another object of the present invention is to provide the use of the said bergamot derivative in the preparation of antibacterial drugs.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a bergamot derivative or a pharmaceutical salt thereof, having the following general structural formula:

[0008]

[0009] L is selected from -(CH2)n-; n is selected from positive integers from 1 to 10;

[0010] R1 is selected from hydrogen, deuterium, hydroxyl, and halogens (fluorine, bromine, chlorine, iodine);

[0011] R2 is selected from hydrogen, deuterium, hydroxyl, and halogens (fluorine, bromine, chlorine, iodine);

[0012] R3 is selected from hydrogen, deuterium, hydroxyl, and halogens (fluorine, bromine, chlorine, iodine);

[0013] R4 is selected from hydrogen, deuterium, hydroxyl, and halogens (fluorine, bromine, chlorine, iodine);

[0014] Selected from R5 is selected from R6 is selected from hydrogen, deuterium, halogens (fluorine, bromine, chlorine, iodine), C1-C10 alkyl, and C1-C10 alkoxy; R7 is selected from hydrogen, deuterium, halogens (fluorine, bromine, chlorine, iodine), C1-C10 alkyl, and C1-C10 alkoxy; R8 is selected from hydrogen, deuterium, halogens (fluorine, bromine, chlorine, iodine), C1-C10 alkyl, and C1-C10 alkoxy; R9 is selected from hydrogen, deuterium, halogens (fluorine, bromine, chlorine, iodine), C1-C10 alkyl, and C1-C10 alkoxy; R 10 Selected from hydrogen, deuterium, halogens (fluorine, bromine, chlorine, iodine), C1-C10 alkyl, and C1-C10 alkoxy groups. Preferably, the bergamot derivative contains:

[0015] L is selected from -(CH2)n-; n is selected from 1, 2, 3, 4, 5;

[0016] R1 is selected from hydrogen, deuterium, hydroxyl, fluorine, bromine, chlorine, and iodine;

[0017] R2 is selected from hydrogen, deuterium, hydroxyl, fluorine, bromine, chlorine, and iodine;

[0018] R3 is selected from hydrogen, deuterium, hydroxyl, fluorine, bromine, chlorine, and iodine;

[0019] R4 is selected from hydrogen, deuterium, hydroxyl, fluorine, bromine, chlorine, and iodine;

[0020] Selected from

[0021] R5 is selected from

[0022] R6 is selected from hydrogen, deuterium, fluorine, bromine, chlorine, iodine, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, tert-butoxy, and n-butoxy.

[0023] R7 is selected from hydrogen, deuterium, fluorine, bromine, chlorine, iodine, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, tert-butoxy, and n-butoxy.

[0024] R8 is selected from hydrogen, deuterium, fluorine, bromine, chlorine, iodine, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, tert-butoxy, and n-butoxy.

[0025] R9 is selected from hydrogen, deuterium, fluorine, bromine, chlorine, iodine, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, tert-butoxy, and n-butoxy.

[0026] R 10 Selected from hydrogen, deuterium, fluorine, bromine, chlorine, iodine, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, tert-butoxy, and n-butoxy.

[0027] Most preferably, the structure of the bergamot derivative is selected from one of the following structures:

[0028]

[0029]

[0030] In a second aspect, the present invention provides the use of the bergamot derivative or a pharmaceutical salt thereof in the preparation of an antibacterial medicament.

[0031] The bacteria referred to are methicillin-resistant Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, etc.

[0032] A third aspect of the present invention provides the use of the bergamot derivative or a pharmaceutical salt thereof in the preparation of a medicament for treating infectious diseases.

[0033] The infectious diseases mentioned refer to those caused by drug-resistant bacteria, such as upper respiratory tract infections, pneumonia, cholecystitis, urinary tract infections, acute tonsillitis, bacterial dysentery, purulent meningitis, scarlet fever, tuberculosis, epidemic cerebrospinal meningitis, folliculitis, boils, etc.

[0034] The drug-resistant bacteria refer to methicillin-resistant Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli.

[0035] In a fourth aspect, the present invention provides the use of the bergamot derivative or a pharmaceutical salt thereof in the preparation of an anti-inflammatory medicament.

[0036] The anti-inflammatory drugs mentioned refer to drugs used to treat leukemia.

[0037] In a fifth aspect, the invention provides the use of the bergamot derivative or a pharmaceutical salt thereof in the preparation of an antitumor medicament.

[0038] The tumor is selected from lung adenocarcinoma or intestinal cancer.

[0039] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0040] The bergamot derivatives provided by this invention are obtained by optimizing the structure of bergamot. Antibacterial activity tests on common Gram-positive bacteria such as drug-resistant Staphylococcus aureus and Gram-negative bacteria such as Escherichia coli and Klebsiella pneumoniae have shown that these compounds have excellent anti-drug-resistant bacterial activity and have great development value in the preparation of anti-infective therapeutic drugs.

[0041] The bergamot derivatives provided by this invention all exhibit good antibacterial activity, and several compounds show excellent antibacterial activity. For example, compound I-4 has a MIC against methicillin-resistant Staphylococcus aureus that is 2 times higher than that of Gepotidacin, and compounds I-5 and I-6 are comparable to Gepotidacin, while bergamot has weaker antibacterial activity. Detailed Implementation

[0042] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1

[0044] Preparation of compound I-1:

[0045]

[0046] Synthesis of 4-(2-bromoethoxy)-7H-furan[3,2-g]thiophene-7-one(3):

[0047] 4-Hydroxy-7H-furano[3,2-g]benzopyran-7-one (compound 1) (50 mg, 0.247 mmol) and anhydrous DMF (7 mL) were added to a 25 mL single-necked flask and stirred until dissolved. Then, 1,2-dibromoethane (compound 2) (278.8 mg, 1.484 mmol) and potassium carbonate (85.4 mg, 0.618 mmol) were added, and the mixture was heated to 100 °C and reacted for 5 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the solvent was removed by vacuum distillation. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by column chromatography to give 38.4 mg of white solid (compound 3), with a yield of 50.3%. 1H NMR(600MHz,Chloroform-d)δ8.26(dd,J=9.8,0.7Hz,1H),7.63(d,J=2.4Hz,1H),7.20(t,J=0.9Hz,1H ),6.92(dd,J=2.4,1.0Hz,1H),6.32(d,J=9.8Hz,1H),4.75–4.70(m,2H),3.73(dd,J=6.2,5.4Hz,2H).

[0048] Synthesis of (1-(7-oxo-7H-furan[3,2-g]furan[3,2-g]furan-4-yl)oxy)ethyl)piperidin-4-ylcarbamate tert-butyl ester (5):

[0049] Compound 3 (38.4 mg, 0.124 mmol) and anhydrous DMF (2.5 mL) were added to a 10 mL single-necked flask. After stirring, 4-(Boc-amino)-piperidine (compound 4) (24.8 mg, 0.124 mmol) and potassium carbonate (42.9 mg, 0.31 mmol) were added, and the mixture was heated to 70 °C and reacted for 2 h. After cooling to room temperature, the mixture was filtered, and the solvent was removed by vacuum distillation. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by column chromatography to give 40.3 mg of white solid (compound 5), with a yield of 75.8%. 1 H NMR(600MHz,Chloroform-d)δ8.22(d,J=9.8Hz,1H),7.63(d,J=2.4Hz,1H),7.20(s,1H),7.02(s,1H),6.32(d,J=9.8Hz,1H),4.55( d,J=64.7Hz,2H),3.75(q,J=7.0Hz,1H),3.53(d,J=16.8Hz,1H),2.93(s,4H),2.33(s,1H),2.01(s,2H),1.63(s,2H),1.47(s,9H).

[0050] Synthesis of 4-(2-(4-aminopiperidin-1-yl)ethoxy)-7H-furan[3,2-g]furan-7-one(6):

[0051] Compound 5 (147 mg, 0.343 mmol) was added to a 10 mL single-necked flask, followed by 2 mL of trifluoroacetic acid and 2 mL of dry dichloromethane. The mixture was reacted at room temperature for 2 h. After the reaction was complete, the solvent was removed by vacuum distillation to obtain 112.6 mg of a yellow oily liquid, which was used directly in the next reaction.

[0052] Synthesis of compound I-1:

[0053] Compound 6 (77.6 mg, 0.236 mmol) and 4-bromo-3,5-difluorobenzaldehyde (compound 7, 52.2 mg, 0.236 mmol) were added to a 10 mL single-necked flask. 2 mL of anhydrous dichloroethane was added and stirred to dissolve the compound. Sodium triacetoxyborohydride (250.1 mg, 1.18 mmol) was then added, and the reaction was allowed to proceed for 3.5 h. After the reaction was complete, saturated NaHCO3 solution was added, and the mixture was extracted with dichloromethane (15 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The product was purified by column chromatography to give 10.5 mg of a white solid, compound I-1, with a yield of 8.3%. 1 H NMR(600MHz,Methanol-d4)δ8.40(dd,J=9.8,0.8Hz,1H),7.82(dd,J=2.3,0.7Hz,1H),7.25–7 .24(m,2H),7.23(d,J=1.8Hz,1H),7.21(d,J=0.9Hz,1H),6.32(dd,J=9.7,0.7Hz,1H),4.70(t, J=5.2Hz,2H),3.98(s,2H),3.36(d,J=0.7Hz,1H),3.25–3.19(m,2H),3.06(t,J=5.3Hz,2H),2. 87–2.80(m,1H),2.44(t,J=11.8Hz,2H),2.09(d,J=13.0Hz,2H),1.64(q,J=12.4,10.6Hz,2H). 13 CNMR(75MHz,MeOD)δ161.74,161.41,158.33,158.18,152.36,148.72,145.53,140.97,140.10,113.75,112.17,1 11.82,111.70,106.73,104.79,96.02,93.34,69.89,56.95,53.66,52.02,30.07,29.36.HRMS(ESI,positive)m / z calcd for C 25 H 23 BrF₂N₂O₄[M+H] + : 533.0887; found 533.0882.

[0054] Example 2

[0055] Preparation of compound I-2:

[0056]

[0057] Following the preparation method of compound I-1 in Example 1, 4-bromo-3,5-difluorobenzaldehyde was replaced with 4-iodo-3-fluorobenzaldehyde to obtain a white solid with a yield of 25.3%. 1 H NMR(600MHz,Methanol-d4)δ8.32(dd,J=9.7,0.7Hz,1H),7.84–7.75(m,2H),7.30 (dd,J=8.9,1.9Hz,1H),7.19(dd,J=2.4,1.0Hz,1H),7.15–7.07(m,2H),6.27(d,J= 9.8Hz,1H),4.64(t,J=5.2Hz,2H),4.05(s,2H),3.19(dd,J=12.0,4.1Hz,2H),3.01 –2.93(m,3H),2.40–2.33(m,2H),2.15–2.09(m,2H),1.68(qd,J=12.1,3.9Hz,2H). 13 C NMR(75MHz,MeOD)δ163.50,161.72,160.25,158.30,152.34,148.72,145.52,140.07,139.69,126.56,116.21,115.88,1 13.78,111.71,106.74,104.78,93.35,80.42,80.08,70.01,56.85,54.28,51.82,29.36,29.19.HRMS(ESI,positive)m / z calcdfor C 25 H 23 F1IN2O4[M+H] + : 563.0843; found 563.0838.

[0058] Example 3

[0059] Preparation of compound I-3:

[0060]

[0061] Following the preparation method of compound I-1 in Example 1, 4-chloro-3,5-difluorobenzaldehyde was substituted for 4-bromo-3,5-difluorobenzaldehyde to obtain a white solid with a yield of 25.2%. 1H NMR(600MHz,Chloroform-d)δ8.22(dd,J=9.8,0.7Hz,1H),7.60(d,J=2.3Hz,1H),7.16(d,J=0.8Hz,1H),7.04–6.97(m,3H),6.29(d,J=9.8Hz,1H),4.55(t, J=5.6Hz,2H),3.80(s,2H),3.02–2.95(m,2H),2.86(t,J=5.6Hz,2H),2.54–2. 51(m,1H),2.27–2.18(m,3H),1.92(d,J=12.8Hz,2H),1.46(q,J=11.4Hz,2H). 13 CNMR(151MHz,DMSO)δ160.63,158.99,157.93,157.35,152.49,149.21,146.61,140.34,114.38,112.84,112. 47,112.47,107.35,105.75,94.17,71.06,57.50,53.85,52.32,48.43,31.72,29.45.HRMS(ESI,positive)m / z calcd for C 25 H 23 ClF₂N₂O₄[M+H] + : 489.1387; found 489.1392.

[0062] Example 4

[0063] Preparation of compound I-4:

[0064]

[0065] Following the preparation method of compound I-1 in Example 1, 4-tert-butoxycarbonylaminopiperidine was replaced with trans-4-(Boc-amino)-3-hydroxypiperidine to obtain a white solid in a yield of 40.5%. 1H NMR(600MHz,Chloroform-d)δ8.21(dd,J=9.8,0.7Hz,1H),7.61(d,J=2.4Hz,1H),7.17(t,J=0.8Hz,1H),7.03–6.9 2(m,3H),6.30(d,J=9.7Hz,1H),4.53(t,J=5.5Hz,2H),3.94(d,J=14.2Hz,1H),3.74(d,J=14.1Hz,1H),3.53–3.46( m,2H),3.18(ddd,J=10.7,4.4,1.8Hz,1H),2.95(d,J=11.8Hz,1H),2.91–2.87(m,1H),2.38(ddd,J=10.5,8.9,4.3 Hz,1H),2.23(td,J=11.6,2.8Hz,3H),2.13(t,J=10.1Hz,1H),2.05(dd,J=13.2,3.7Hz,1H),1.44–1.33(m,1H).13C NMR (151MHz, CDCl3) δ161.20,160.77,159.11,158.13,152.58,148.69,145.07,142.76,139.42,113.92,112.71,111. 35,111.19,107.26,104.78,94.41,70.82,70.76,60.81,58.91,57.35,52.71,49.65,29.46.HRMS(ESI,positive)m / z calcd for C 25 H 23 BrF₂N₂O₅[M+H] + : 549.0836; found 549.0831.

[0066] Example 5

[0067] Preparation of compound I-5:

[0068]

[0069] Following the preparation method of compound I-1 in Example 1, 4-tert-butoxycarbonylaminopiperidine and 4-bromo-3,5-difluorobenzaldehyde were replaced with trans-4-(Boc-amino)-3-hydroxypiperidine and 4-iodo-3-fluorobenzaldehyde to obtain a white solid in a yield of 10.1%. 1HNMR(600MHz,Chloroform-d)δ8.21(dd,J=9.7,0.7Hz,1H),7.70(ddd,J=15.5,8.1,6.5Hz,1H),7.61(d,J=2.4Hz,1H),7.17( t,J=0.8Hz,1H),7.11(dt,J=8.9,2.3Hz,1H),6.96(dd,J=2.3,1.0Hz,1H),6.91(dt,J=8.2,2.3Hz,1H),6.29(s,1H),4.68(s,1 H),4.52(t,J=5.5Hz,2H),3.95(d,J=13.7Hz,1H),3.75(s,1H),3.49(td,J=9.2,4.4Hz,1H),3.18(ddd,J=10.7,4.5,1.8Hz,1H ),2.97–2.93(m,1H),2.93–2.82(m,2H),2.39(ddd,J=10.7,8.8,4.3Hz,1H),2.21(td,J=11.5,2.8Hz,1H),2.16–2.01(m,2H). 13 C NMR (151MHz, CDCl3) δ162.63,161.22,158.13,152.59,148.69,145.08,139.32,125.45,123.85,115.46,113.96,112.75 ,107.30,104.78,94.45,70.82,70.55,63.90,60.79,58.89,57.33,52.73,49.59,29.27.HRMS(ESI,positive)m / zcalcd for C 25 H 24 FIN2O5[M+H] + : 579.0792; found 579.0787.

[0070] Example 6

[0071] Preparation of compound I-6:

[0072]

[0073] Following the preparation method of compound I-1 in Example 1, 4-tert-butoxycarbonylaminopiperidine and 4-bromo-3,5-difluorobenzaldehyde were replaced with trans-4-(Boc-amino)-3-hydroxypiperidine and 4-chloro-3,5-difluorobenzaldehyde to obtain a white solid with a yield of 39.3%. 1H NMR(600MHz,Chloroform-d)δ8.21(dd,J=9.8,0.7Hz,1H),7.61(d,J=2.3Hz,1H),7.18(t,J=0.8Hz,1H),7.0 5–7.01(m,3H),6.97(dd,J=2.4,1.0Hz,1H),6.31(d,J=9.8Hz,1H),4.69(t,J=0.8Hz,1H),4.53(t,J=5.5Hz,2 H),3.95(d,J=14.1Hz,1H),3.75(d,J=14.1Hz,1H),3.21–3.16(m,1H),2.95(d,J=11.8Hz,1H),2.93–2.85(m ,2H),2.42–2.35(m,1H),2.23(td,J=11.4,2.7Hz,1H),2.13(t,J=10.1Hz,1H),2.06(dq,J=13.0,3.6Hz,2H). 13 C NMR (151MHz, CDCl3) δ161.19,158.14,152.60,148.66,145.10,139.38,113.99,112.80,111.49,111.35, 107.33,104.75,94.50,70.77,60.76,58.84,57.33,52.64,50.85,49.53,29.68.HRMS(ESI,positive)m / z calcd for C 25 H 23 CIF2N2O5[M+H] + : 505.1342; found 505.1336.

[0074] Example 7

[0075] Preparation of compound I-7:

[0076]

[0077] Following the preparation method of compound I-1 in Example 1, 4-tert-butoxycarbonylaminopiperidine and 4-bromo-3,5-difluorobenzaldehyde were replaced with trans-4-(Boc-amino)-3-hydroxypiperidine and 3,4-dihydro-2H-pyrano[2,3-C]pyridine-6-carboxaldehyde to obtain a white solid in 39.3% yield. 1H NMR(600MHz,Chloroform-d)δ8.22(dd,J=9.7,0.7Hz,1H),7.61(d,J=2.4Hz,1H),7.15(t,J=0.8Hz,1H),6.98–6 .93(m,2H),6.90(dd,J=8.3,2.1Hz,1H),6.84(d,J=8.2Hz,1H),6.35(d,J=9.7Hz,1H),5.31(s,1H),4.49(t,J=5. 5Hz,2H),4.23(s,4H),4.04(d,J=13.1Hz,1H),3.79–3.68(m,2H),3.22–3.17(m,1H),2.98(d,J=11.7Hz,1H),2. 85(qt,J=13.6,5.5Hz,2H),2.54(d,J=7.3Hz,1H),2.20–2.13(m,1H),2.08(t,J=10.4Hz,2H),1.74–1.66(m,1H). 13 C NMR (151MHz, CDCl3) δ161.27,158.05,152.51,148.58,145.10,143.61,143.45,139.47,129.10,121.92,117.79,117.53,113.96,11 2.75,107.27,104.81,94.35,70.78,69.16,64.28,60.36,59.10,57.06,53.44,52.38,49.18,27.88.HRMS(ESI,positive)m / zcalcd for C 27 H 28 N₂O₇[M+H] + : 493.1975; found 493.1969.

[0078] Example 8

[0079] In vitro antibacterial activity assay of compounds I-1 to I-7

[0080] The in vitro antibacterial activity of three strains—methicillin-resistant Staphylococcus aureus ATCC 33591 (MRSA), Klebsiella pneumoniae ATCC 10031, and Escherichia coli ATCC 25922—was tested using the micro-broth dilution method. Gepotidacin and bergamot were used as positive controls, and the minimum inhibitory concentration (MIC) was determined.

[0081] The experimental strains included Gram-positive bacteria: methicillin-resistant Staphylococcus aureus ATCC 33591 (MRSA); and Gram-negative bacteria: Klebsiella pneumoniae ATCC 10031 and Escherichia coli ATCC 25922.

[0082] The specific operating steps are as follows:

[0083] (1) Preparation of test compound and culture medium: The test compound was prepared as a 10 mg / mL DMSO stock solution, and the culture medium was prepared according to the manufacturer's instructions.

[0084] (2) Gradual dilution of the test compound

[0085] Two-fold dilution of the test compound solution: Aseptically, add 12.8 μL of the drug solution to well 1 of a 96-well plate, then add 27.2 μL of DMSO. Add 2-12 μL of DMSO to each of the 20 wells and mix thoroughly. Take 20 μL from the first well and add it to the second well, mixing thoroughly. Collect 20 μL from the second well and add it to the third well, repeating this process until the eleventh well. Collect 20 μL and discard it. From the twelfth well onwards, add only 20 μL of DMSO for growth and control. Add 180 μL of sterile water to the next well and mix thoroughly. The control drug is prepared using the same method. Prepare different concentrations of the test compound solution as samples by placing 10 μL in each of the 96 sterile wells.

[0086] (3) Preparation of test strains and MIC experiment

[0087] Agar plates containing a single fungus were mixed with 5 ml of sterile physiological saline, and the spectrophotometer transmittance was approximately 80%. MHB liquid medium was diluted to 1:200, and 90 μL of the bacterial suspension was mixed with 10 μL of the target compound solution in the medium. Results were incubated at 35°C for 16–20 hours and then interpreted. Dilutes of the same compound were recorded in wells 1 through 12 as follows: 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125, and 0 μg / mL. Each dilution of the same compound was used as a parallel sample.

[0088] (4) Results reading and judgment:

[0089] Referring to the 2013 CLSI recommendations: the MIC value is the lowest visible antimicrobial concentration that can completely inhibit the growth of microorganisms in a test tube or microdilution well. If only microorganisms growing in the pores can be identified, the device can be used to help read and record the results of microdilution experiments. When determining the growth endpoint, compare the growth of microorganisms in microdilution wells or in vitro with those of non-antimicrobial agents. The test is valid if the growth control wells show acceptable growth conditions (2mm button-shaped precipitates or a mixture with a certain degree of turbidity).

[0090] Table 1. Minimum inhibitory concentration (MIC, μg / mL) of compound I-1-I-7 against three bacterial strains.

[0091]

[0092]

[0093] As shown in Table 1, all compounds exhibited good antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). Compounds I-5 and I-6 showed comparable activity to the positive control drug Gepotidacin, while compound I-4 exhibited twice the antibacterial activity compared to Gepotidacin, with a MIC of 0.25 μg / mL. All compounds also demonstrated good antibacterial activity against Klebsiella pneumoniae, with compounds I-1, I-2, and I-3 having MICs of 1 μg / mL, 0.5 μg / mL, and 1 μg / mL, respectively. Compounds I-1, I-2, and I-3 also showed some inhibitory activity against Escherichia coli. These results indicate that the compounds of this invention have the potential for developing anti-infective drugs.

[0094] Example 9

[0095] In vitro anti-inflammatory activity assays of some compounds of this invention

[0096] Mouse mononuclear macrophage leukemia cell line RAW264.7 (purchased from Wuhan Pronosei Life Sciences Co., Ltd.) was selected as the cell line for anti-inflammatory activity assay. The expression levels of inflammatory factors such as TNF-α and IL-6 were evaluated. The anti-inflammatory activity was assessed using an inflammation-related physiological index (IR) score, ranging from 0 to 100. The IR scoring method was as follows: Cell-only culture medium was used as the blank group. Lipopolysaccharide (LPS) was added to the blank group to induce the expression of inflammatory factors in cells that did not exhibit inflammation, creating the model group. The compound was added to the model group as the experimental group. Inflammation-related index levels were measured in each group, and the results were calculated using the formula: IR = (Inflammation-related index level in model group - Inflammation-related index level in experimental group) / (Inflammation-related index level in model group - Inflammation-related index level in blank group) × 100%. The IR calculation formula used enzyme-linked immunosorbent assay (ELISA) to quantitatively characterize the inflammation-related index levels by detecting absorbance.

[0097] The specific experimental method was as follows: RAW264.7 cells were cultured in DMEM medium until they reached the logarithmic growth phase. After cell counts, they were seeded into 96-well cell culture plates. Once the cells were fully adherent, the old medium was discarded. 0.5 μM and 1 μM solutions of the compound prepared with the medium, along with the positive control compound 8-hydroxypsoralen, were added to the 96-well plates, and the plates were incubated for 2 hours. Lipopolysaccharide (LPS) was then added to induce the secretion of inflammatory factors for 4 hours. The cell supernatant was collected, and the levels of inflammatory factors such as TNF-α and IL-6 were measured according to the instructions of the enzyme-linked immunosorbent assay (ELISA) kit.

[0098] Table 2 shows the inhibitory effect of some of the compounds in the examples on IL-6 in RAW264.7 cells (5 μM).

[0099]

[0100]

[0101] Table 3 shows the inhibitory effect of some of the compounds in the examples on TNF-α in RAW264.7 cells (5 μM).

[0102]

[0103] As shown in Tables 2 and 3, the compounds of this invention exhibit good inhibitory effects on IL-6 and TNF-α in mouse mononuclear macrophage leukemia RAW264.7. For example, all the compounds showed better inhibitory effects on IL-6 than bergamot, and compounds I-6 and I-7 also showed better inhibitory effects on TNF-α than bergamot. These results indicate that the compounds of this invention can also be used to prepare anti-inflammatory drugs.

[0104] Example 10

[0105] In vitro antitumor activity assay of the compounds of this invention

[0106] Tumor cell proliferation inhibition assays were performed on some of the compounds of the present invention using the CCK-8 assay (e.g., Lü Qiujun, ed., New Drug Pharmacological Research Methods, 2007: 242-243).

[0107] The cell lines used were A549 (human lung adenocarcinoma cells) and HCT-116 (human colon cancer cells), and the culture medium was DMEM + 10% FBS + double antibiotics.

[0108] In vitro activity assay: 4-5 × 10⁻⁵ mg / well of the solution was added to each well of a 96-well plate. 4 100 μL of cell suspension per cell / mL was incubated at 37°C in a 5% CO2 incubator. After 24 h, 10 μL of sample solution was added to each well, with duplicate wells, and incubated at 37°C in a 5% CO2 incubator for 72 h. 20 μL of 5 mg / mL CCK-8 solution was added to each well, and after 4 h, 100 μL of dissolving buffer was added to each well. The wells were then incubated to dissolve the cells, and the OD value at 570 nm was measured using a multi-wavelength microplate reader. The inhibition rate (IR) and half-maximal inhibitory concentration (IC50) were calculated using Excel software. 50 ).

[0109] Calculate the inhibition rate (IR%) of cell growth by the drug using the following formula.

[0110]

[0111] The experimental results are shown in Table 4, where the sample refers to the compound prepared in the corresponding example.

[0112] Table 4 shows the in vitro antitumor activity of compounds in some examples.

[0113]

[0114] The above experimental results show that the compounds of the present invention have good antitumor activity, and exhibit good activity against lung cancer, colorectal cancer, and other cell lines. For example, compounds I-1 and I-2 show activity of several micromoles against A549 tumor cell line, while bergamot has virtually no antitumor activity. Therefore, the compounds of the present invention and their salts can be used to prepare antitumor drugs.

[0115] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A bergapten derivative or a pharmaceutically acceptable salt thereof, characterized by, The structural general formula is as follows: L is selected from -(CH2)n-; n is selected from 1; R1 is selected from hydrogen, hydroxyl; R2 is selected from hydrogen; R3 is selected from hydrogen; R4 is selected from hydrogen; selected from R5is selected from R6 is selected from hydrogen, halogen; R7 is selected from hydrogen, halogen; R8 is selected from hydrogen, halogen; R9 is selected from hydrogen, halogen; R 10 selected from hydrogen, halogen.

2. The perillyl alcohol derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein The structure of the nerolidol derivative is selected from one of the following structures:

3. Use of the nerolidol derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 in the preparation of an antibacterial medicament.

4. Use of a nerolidol derivative or a pharmaceutical salt thereof according to claim 3 in the manufacture of an antibacterial medicament, characterized in that, The bacteria refer to methicillin-resistant Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli.

5. Use of the nerolidol derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 in the preparation of a medicament for treating an infectious disease.

6. Use of the nerolidol derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 in the preparation of an anti-inflammatory medicament.

7. Use of the nerolidol derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2 in the preparation of an anti-tumor medicament.

8. Use of the perillyl alcohol derivative or a pharmaceutical salt thereof according to claim 7 for the manufacture of an antitumor agent, characterized in that, The tumor is selected from lung adenocarcinoma or intestinal cancer.

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