A macrolide derivative, a process for its preparation and its use

Abstract

The application provides a macrolide derivative and a preparation method and application thereof, relates to the technical field of medicinal chemistry, and the macrolide derivative is a compound with a general structure shown in the following formula I: in the formula, A is selected from any one of alkynyl, alkenyl or piperazine; R is selected from any one of H or F; X is selected from any one of CH or N, Y is selected from any one of methyl, ethyl or cyclopropyl, and n is an integer in the range of 2-6. The macrolide derivative is connected by different lengths of ether hydrocarbons at the 3-position, thereby avoiding the problem that the instability of a synthesized compound is caused by using an ester group as a side chain to connect a quinolone group. The application successfully prepares a macrolide derivative with double targets, and the antibacterial activity of the macrolide derivative is obviously better than the antibacterial activity of erythromycin, telithromycin, clarithromycin and ciprofloxacin which only have single targets.

Description

Technical Field

[0001] This application relates to the field of medicinal chemistry, and in particular to a macrolide derivative, its preparation method, and its application. Background Technology

[0002] Macrolide antibiotics, such as erythromycin, have long been important drugs for treating a variety of infections caused by pathogenic microorganisms, especially in the treatment of upper and lower respiratory tract infections and skin and soft tissue infections. Due to their mild side effects, these drugs have long provided a highly effective and safe treatment option in clinical practice. However, with the long-term use of antibiotics, the problem of drug resistance in clinical practice has become increasingly serious. For example, common pathogens such as Streptococcus pneumoniae, Staphylococcus aureus, and Streptococcus pyogenes have shown a significant increase in resistance to first-generation macrolide antibiotics such as erythromycin.

[0003] To address this issue, second-generation macrolide antibiotics such as clarithromycin and azithromycin were developed. While these drugs exhibit high acid resistance and favorable pharmacokinetic properties, they also lack antibacterial activity against erythromycin-resistant bacteria. This is primarily because their shared cladinose structure readily induces bacterial resistance, leading to modification of the key antibacterial target—the ribosomal base A2058—by methyltransferases expressed by the erm gene, resulting in high levels of resistance. Furthermore, the resistance mechanism also involves efflux pump proteins expressed by mef cells expelling erythromycin-containing compounds containing cladinose from the cell.

[0004] To overcome these resistance problems, third-generation erythromycin ketone lactones, such as telithromycin, have been developed. Telithromycin exhibits good antibacterial activity against constitutively resistant Streptococcus pneumoniae and Streptococcus pyogenes by removing cladinose and oxidizing the 3-OH group to a glycosyl group, simultaneously acting on the base pairs A752 and U2609 of the microbial ribosome. However, telithromycin is ineffective against constitutively resistant Staphylococcus aureus and drug-resistant Mycoplasma, and its hepatotoxicity limits its clinical use.

[0005] In recent years, in-depth research on ribosome structure has revealed that the ribosomal base pairs C1782-C2586 are evolutionarily conserved sites, and are very close to the binding site of erythromycin, potentially becoming new targets for enhancing activity against drug-resistant bacteria. This discovery provides new insights for the development of novel macrolide antibiotics.

[0006] Existing macrolide antibiotics mostly target a single antibacterial site, which limits their antibacterial efficacy and ability to combat drug resistance. Therefore, developing macrolide derivatives with dual targets, capable of acting on multiple key sites simultaneously, holds promise for further improving their antibacterial properties and reducing the development of drug resistance.

[0007] However, current research on 3-position derived side chains mainly focuses on ester groups. While these derivatives exhibit antibacterial activity against induced and efflux resistant bacteria, they are difficult to obtain activity against constitutively resistant bacteria and are easily hydrolyzed by intravenous enzymes and gastric acid. Therefore, there is an urgent need to develop novel macrolide antibiotics with 3-position ether side chains to address these issues. Summary of the Invention

[0008] This application provides a macrolide derivative, its preparation method, and its application to solve the above-mentioned problems.

[0009] In a first aspect, this application provides a macrocyclic lactone derivative, which is a compound having the general formula shown in Formula I:

[0010]

[0011] In the formula, A is selected from any one of alkynyl, alkenyl or piperazine; R is selected from any one of H or F; X is selected from any one of CH or N; Y is selected from any one of methyl, ethyl or cyclopropyl; and n is an integer ranging from 2 to 6.

[0012] In one optional embodiment of this application, A is selected from piperazine or alkynyl, R is selected from F, X is selected from CH or N, Y is selected from ethyl or cyclopropyl, and n is an integer ranging from 3 to 5.

[0013] In one optional embodiment of this application, A is selected as piperazine, R is selected as F, X is selected as N, Y is selected as ethyl, and n is 3.

[0014] A is selected as piperazine, R as F, X as CH, Y as cyclopropyl, and n is 5.

[0015] In one optional embodiment of this application, the structure of the macrocyclic lactone derivative as described in any one of the first aspects above is as follows:

[0016]

[0017] In a second aspect of this application, a method for preparing a macrocyclic lactone derivative is provided. The method includes: introducing different side chains into the compound shown in formula (1), introducing a quinolone group through a reaction, and removing the protecting group to obtain the compound shown in formula (I).

[0018]

[0019] In the formula, A, R, X, Y, and n are defined independently of each other as described in any one of the first aspects above.

[0020] In one optional embodiment of this application, the method includes a first synthesis method, which is used to synthesize a compound of formula II in which A is selected as an alkynyl group, R is selected as H, X is selected as CH, and Y is selected as methyl.

[0021]

[0022] In the first synthesis method, the compound shown in formula (1) is introduced with alkynol side chains of different lengths to obtain the compound shown in formula (2). The compound shown in formula (2) is introduced with quinolone groups through a sonogashira coupling reaction. After removing the protecting group, the compound shown in formula II is obtained. In the compound shown in formula II, the value of n is an integer ranging from 2 to 6.

[0023] In one optional embodiment of this application, the method includes a second synthesis method, which is used to synthesize a compound of formula III where A is selected as alkenyl, R as H, X as CH, Y as methyl, and n is 4.

[0024]

[0025] In the second synthesis, the compound shown in formula (1) is introduced with an enol side chain to obtain the compound shown in formula (3). The compound shown in formula (3) is introduced with a quinolone group through a Heck coupling reaction, and after removing the protecting group, the compound shown in formula III is obtained.

[0026] In one optional embodiment of this application, the method includes a third synthesis method for synthesizing a compound of formula IV, where A is selected as piperazine and R is selected as F.

[0027]

[0028] In the third synthesis, the compound shown in formula (1) is introduced with diol side chains of different lengths to obtain the compound shown in formula (4). The compound shown in formula (4) is sulfonated and then a quinolone group is introduced through a nucleophilic substitution reaction. After removing the protecting group, the compound shown in formula IV is obtained. In the compound shown in formula IV, X, Y, and n independently have the definitions of any one of the first aspects above.

[0029] In a third aspect of this application, there is provided the use of at least one of the macrolide derivatives, isotopic labels thereof, solvates, polymorphs, pharmaceutically acceptable salts thereof, or prodrug compounds as described in any of the first aspects above in the preparation of a medicament, wherein the medicament is an antimicrobial drug and the pathogenic microorganism is a bacterium.

[0030] In one optional embodiment of this application, at least one of the macrolide derivatives, isotopic labels thereof, solvates, polymorphs, pharmaceutically acceptable salts thereof, or prodrug compounds as described in any of the first aspects above is used in the preparation of antibiotics.

[0031] This application offers the following advantages: It provides a macrolide derivative, its preparation method, and its application. The macrolide 3-position ether hydrocarbon derivative is prepared by linking a quinolone group of different lengths to the 3-position of the macrolide. The macrolide derivative prepared in this application exhibits dual-target activity, inhibiting protein synthesis and DNA replication, and can act on multiple key sites simultaneously. Its antibacterial activity is significantly improved compared to single-target derivatives such as erythromycin, telithromycin, clarithromycin, and ciprofloxacin. Furthermore, compared to disclosed macrolide 3-position ester-modified derivatives, the macrolide derivative prepared in this application not only shows higher antibacterial activity against induced and efflux-resistant bacteria but also successfully restores antibacterial activity against constitutively ERM-resistant bacteria.

[0032] In addition, the macrocyclic lactone derivatives prepared in this application improve the stability of the compound and avoid the problem that the synthesized compound is unstable under acidic conditions due to the use of ester groups as side chains to connect quinolone groups. Attached Figure Description

[0033] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments of this application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0034] Figure 1 This is a diagram showing the results of a DNA gyrase supercoiling experiment of a target compound 33b provided in an embodiment of this application;

[0035] Figure 2 This application provides an embodiment of the target compound 33b with inhibitory activity against protein synthesis (IC). 50 The measurement results are shown in the figure. Detailed Implementation

[0036] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0037] Erythromycin is an important macrolide antibiotic used clinically to treat infections of the upper and lower respiratory tracts, as well as skin and soft tissue infections caused by pathogenic microorganisms. It has mild side effects and has provided a highly effective and safe route of administration for humans, especially children, for half a century. However, long-term use has led to serious clinical resistance, such as against Streptococcus pneumoniae, Staphylococcus aureus, and Streptococcus pyogenes. Second-generation erythromycins—clarithromycin and azithromycin—which emerged in the 1980s, while exhibiting high acid resistance and good pharmacokinetic properties, also lack antibacterial activity against erythromycin-resistant bacteria. The main reason is that their shared substructure, the claridin sugar at position 3, induces bacterial resistance, leading to high levels of resistance due to modification of the key antibacterial target—the ribosomal base A2058—by methyltransferases expressed by the erm gene. In addition, drug resistance mechanisms also include the efflux pump proteins expressed by mef cells, which pump erythromycin-like compounds containing cladinose out of the cell.

[0038] As a third-generation erythromycin ketolactone, telithromycin removes the cladinose and oxidizes the 3-OH group to a glycosyl group. Simultaneously, its 11-position side chain interacts with the A752 and U2609 base pairs of the microbial ribosome, thus exhibiting good antibacterial activity against constitutively resistant Streptococcus pneumoniae and Streptococcus pyogenes. It is the only erythromycin derivative approved for the treatment of community-acquired bacterial pneumonia to date. However, it has no activity against constitutively resistant Staphylococcus aureus and drug-resistant Mycoplasma. Ketolactones in phase III clinical trials, such as solithromycin, share the same antibacterial target as telithromycin. After its market launch, telithromycin was found to have hepatotoxicity, leading to strict restrictions on its use. Other compounds in clinical trials, due to their similar ketolactone structures, have faced safety concerns, resulting in telithromycin being the only erythromycin approved for marketing over the past two decades.

[0039] Recent findings indicate that the ribosomal base pairs C1782-C2586 are evolutionarily conserved sites, very close to the erythromycin binding site. Computer simulations and cryo-electron microscopy structural analysis of the complex revealed it could serve as a novel target for erythromycin derivatives to enhance anti-drug-resistant bacterial activity. Therefore, removing cladinose and introducing a side chain can not only eliminate induced resistance but also target different previously reported anti-drug-resistant bacterial targets. Currently, side chains derived at the 3-position are primarily derived via the ester group at the 3-OH position, such as acyl lactones, which are easily hydrolyzed by in vivo enzymes and gastric acid. Structure-activity relationships show that 3-ester side chain derivatives possess antibacterial activity against induced and efflux resistant bacteria, but rarely exhibit activity against constitutively resistant bacteria; stereoisomerism at C-3 leads to loss of antibacterial activity.

[0040] Therefore, in response to the current problem of drug resistance in macrolide antibiotics, this application aims to develop a novel macrolide antibiotic with dual targets and a 3-position ether side chain, in order to improve its antibacterial activity against drug-resistant bacteria and provide a new drug option for the clinical treatment of drug-resistant bacterial infections.

[0041] The following describes a macrocyclic lactone derivative, its preparation method, and its application, with specific embodiments. Unless otherwise stated, the raw materials and reagents used in the following embodiments are commercially available or can be prepared by known methods. Regarding the synthesis method of the target compound, this application can use a stepwise introduction method, i.e., first introducing alkynol side chains of different lengths, then introducing a quinolone group through a Sonogashira coupling reaction, and finally removing the protecting group to obtain the target compound; or first introducing an enol, then introducing a quinolone group through a Heck coupling reaction, and finally removing the protecting group to obtain the target compound; or introducing diol side chains of different lengths, then sulfonating them, reacting with a quinolone reagent, and finally removing the protecting group to obtain the compound of this application. The synthetic route is briefly described below:

[0042]

[0043] Preparation Example 1: Intermediate Compound 1

[0044] Intermediate compound 1 is a known compound, and the synthetic method of compound 1 can be found in European Journal of Medicinal Chemistry 59(2013)54-63;

[0045]

[0046] Preparation Example 2: Intermediate Compound 2 and Intermediate Compound 3

[0047]

[0048] Reaction conditions and reagents: a. DMS, NCS, triethylamine, DCM, -10℃, 6h; b. Sodium borohydride, dioxane, rt, 8h.

[0049] Synthesis of intermediate compound 2:

[0050]

[0051] NCS (1.459 g, 10.93 mmol) was placed in a dry 250 mL round-bottom flask, and dry DCM (50 mL) was added. The flask was placed in an ice-salt bath and stirred until homogeneous. DMS (0.95 mL, 12.98 mmol) was then added, resulting in a white flocculent precipitate. The flask was purged with argon gas, sealed, and stirred for 30 min. Dry compound 1 (4.692 g, 6.83 mmol) was placed in a dry constant-pressure dropping funnel and dissolved in dry DCM (50 mL) and slowly added dropwise. The reaction was allowed to proceed for approximately 3-4 h, and the reaction was monitored by TLC. Triethylamine (0.70 mL, 13.66 mmol) was then added, and the reaction was allowed to proceed for approximately 0.5-1 h, with the reaction being monitored by TLC. After the reaction was complete, the organic phase was washed once each with saturated NaHCO3 solution, distilled water, and saturated NaCl solution. The solvent was removed under reduced pressure, and the mixture was purified by column chromatography. The column chromatography conditions were: 100-200 mesh silica gel, and the mobile phase was dichloromethane / ethanol / ammonia water = 10 / 0.1 / 0.05. This yielded a pale yellow, fluffy solid intermediate compound 2 (2.000 g, 2.92 mmol, 42.8%).

[0052] Synthesis of intermediate compound 3:

[0053]

[0054] The obtained intermediate compound 2 (2.000 g, 2.92 mmol) was dissolved in dioxane (15 mL), stirred until homogeneous, and sodium borohydride (0.530 g, 14.02 mmol) was added. An argon balloon was connected via a three-way valve, and multiple purgings were performed to ensure the reaction flask was filled with argon gas. The flask was then sealed. The reaction was carried out at room temperature for approximately 8-9 hours, and the reaction progress was monitored by TLC. After the reaction was complete, the organic phase was washed once with distilled water and once with saturated NaCl solution. The solvent was removed under reduced pressure, and the mixture was purified by column chromatography. The column chromatography conditions were: 100-200 mesh silica gel, and the mobile phase was dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05. This yielded a white, fluffy solid intermediate compound 3 (0.800 g, 1.17 mmol, 40.1%). HRMS (ESI) (M+H) + m / z 687.4022, calcd for C 34 H 59 N2O 12 687.4063. 1H NMR (CDCl3, 400MHz) δ: 5.21 (dd, J1 = 10.4Hz, J2 = 2.8Hz, 1H), 4.90 (s, 1H), 4.78 (dd, J1 = 10.4Hz, J2 = 8.0Hz, 1H), 4.42 (d, J = 1.0Hz, 1H), 4 .25(d,J=7.5Hz,1H),3.84(s,3H),3.75-3.68(m,1H),3.57-3.51(m,1H),3.27-3.20(m,1H),2.99(s,3H),2.97-2.89(m,1H),2.86(d,J =10.7Hz,1H),2.78-2.65(m,1H),2.50(q,J=6.8Hz,1H),2.31(s,6H),2.08(s,3H),1.99-1.90(m,1H),1.81-1.70(m,2H),1.65-1.55(m ,1H),1.53(s,3H),1.40(d,J=7.1Hz,3H),1.36(s,3H),1.34-1.29(m,2H),1.28-1.23(m,6H),0.98-0.93(m,6H),0.90(t,J=7.3Hz,3H).

[0055] Preparation Example 3: Preparation of intermediate compound 4-19

[0056]

[0057] Reaction conditions and reagents: b. at-BuONa, acrylonitrile, ice-water bath, 1 h; c. platinum dioxide, acetic acid, H2, rt, 2 days; d. 3-butyn-1-ol / 4-pentyn-1-ol / 5-hexyn-1-ol / 6-heptyyn-1-ol / 7-octyyn-1-ol, formic acid, sodium nitrite, -10℃, 1 week; e. 5-hexen-1-ol, formic acid, sodium nitrite, -10℃, 1 week; f. 1,2-ethylene glycol / 1,3-propanediol / 1,4-butanediol / 1,5-pentanediol, formic acid, sodium nitrite, -10℃, 1 week; g. p-toluenesulfonyl chloride, DMAP, triethylamine, DCM, rt, 2 h.

[0058] Preparation Example 3.1: General Synthetic Method for Intermediate Compounds 4 and 5

[0059] The obtained intermediate compound 1 or 3 (1 eq) was dissolved in acrylonitrile (150 eq), placed in an ice-water bath, and sodium tert-butoxide (1 eq) was added. The reaction was carried out for about 1 h, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was filtered, and the filter cake was washed with DCM. The filtrate was washed once with water and once with saturated NaCl solution. The intermediate compounds 4 and 5 were purified by organic phase rotation dry column chromatography.

[0060] Synthesis of intermediate compound 4

[0061]

[0062] Intermediate compound 1 (4.085 g, 6.09 mmol) was dissolved in acrylonitrile (60 mL, 913.50 mmol), stirred thoroughly in an ice-water bath, and sodium tert-butoxide (0.585 g, 6.09 mmol) was added. Following the general synthetic method of this preparation example, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give intermediate compound 4 (3.985 g, 5.39 mmol, 88.5%), a pale yellow, fluffy solid. HRMS(ESI)(M+H) + m / z 740.4333, calcd for C 37 H 62 N3O 12 740.4328. 1 H NMR (CDCl3, 400MHz) δ: 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.82 (s, 1H), 4.76 (dd, J1 = 10.6Hz, J2 = 7.4Hz, 1H), 4.45 (d, J = 7.4Hz, 1H), 3.96-3.90 (m, 1H ),3.84(s,3H),3.82-3.78(m,1H),3.73-3.67(m,1H),3.65(d,J=3.9Hz,1H) ,3.56-3.48(m,1H),3.26(d,J=10.5Hz,1H,2.98(s,3H),2.91-2.76(m,2H), 2.71-2.63(m,2H),2.47(q,J=6.9Hz,1H),2.28(s,6H),2.07(s,3H),1.97- 1.86(m,2H),1.80-1.75(m,1H),1.62-1.53(m,1H),1.49(s,3H),1.42-1.36 (m,1H),1.34(s,3H),1.31(d,J=6.8Hz,3H),1.29-1.22(m,7H),1.21-1.15( m,1H),0.96(d,J=7.0Hz,3H),0.92(d,J=7.6Hz,3H),0.88(t,J=7.4Hz,3H).

[0063] Synthesis of intermediate compound 5

[0064]

[0065] Intermediate compound 3 (0.800 g, 1.17 mmol) was dissolved in acrylonitrile (11 mL, 175.50 mmol), stirred thoroughly in an ice-water bath, and sodium tert-butoxide (0.112 g, 1.17 mmol) was added. Following the general synthetic method of this preparation example, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give intermediate compound 5 (0.560 g, 0.76 mmol, 65.0%), a pale yellow, fluffy solid. HRMS(ESI)(M+H) + m / z 740.4313, calcd for C 37 H 62 N3O 12 740.4328. 1 H NMR (CDCl3, 400MHz) δ: 5.07 (dd, J1 = 8.9Hz, J2 = 3.6Hz, 1H), 4.95 (s, 1H), 4.78 (dd, J1 = 10.5Hz, J2 = 7.4Hz, 1H), 4.44 (d, J = 7.5Hz, 1H), 4.05(s,1H),3.98-3.89(m,1H),3.83(s,3H),3.78-3.64(m,2H),3.61-3.50(m,1H),3.31(br,1H),3.07-2.91(m,4H),2.89-2.62(m,3 H),2.54(q,J=6.9Hz,1H),2.31(s,6H),2.25-2.15(m,1H),2.08(s,3H),2.00-1.88(m,1H),1.84-1.75(m,1H),1.60-1.54(m,1H),1. 51(s,3H),1.48-1.36(m,2H),1.36-1.31(m,6H),1.31-1.27(m,4H),1.25(d,J=6.7Hz,3H),1.03(d,J=7.1Hz,3H),0.99-0.90(m,6H).

[0066] Preparation Example 3.2: General Synthetic Method for Intermediate Compounds 6 and 7

[0067] The obtained intermediate compounds 4 and 5 (1 eq) were dissolved in acetic acid (10 times the mass of intermediate compounds 4 and 5), and platinum dioxide (0.1 times the mass of intermediate compounds 4 and 5) was added. A high-pressure reactor was used, and the reactor was repeatedly purged with hydrogen to ensure it was completely filled. The reaction was carried out at room temperature for approximately 4-6 hours, and the reaction progress was monitored by TLC. After the reaction was complete, the mixture was filtered through diatomaceous earth. The filtrate was adjusted to pH 9-10 with 2M NaOH solution, and the organic phase was washed once with distilled water and once with saturated NaCl solution. The organic phase was then purified by rotary cyclohexane column chromatography to obtain intermediate compounds 6 and 7.

[0068] Synthesis of intermediate compound 6

[0069]

[0070] Intermediate compound 4 (3.985 g, 5.39 mmol) was dissolved in acetic acid (40 mL, 10 times the mass of intermediate compound 4) and stirred until homogeneous. Platinum dioxide (0.399 g, 0.1 times the mass of intermediate compound 4) was then added. Following the general synthetic method of this preparation example, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.6 / 0.1) to give a white, fluffy solid intermediate compound 6 (2.085 g, 2.80 mmol, 52.0%). HRMS (ESI) (M+H) + m / z 744.4636, calcd for C 37 H 66 N3O 12 744.4641. 1 H NMR (CDCl3, 400MHz)

[0071] δ:5.15(dd,J1=10.8Hz,J2=2.5Hz,1H),4.85(s,1H),4.77(dd,J1=10.5Hz,J2

[0072] =7.4Hz,1H),4.45(d,J=7.5Hz,1H),3.84(s,3H),3.82-3.74(m,1H),3.73-3.62(m,3H),3.54-3.42( m,1H),3.19(d,J=10.5Hz,1H),2.98(s,3H),2.92-2.68(m,4H),2.47(q,J=6.8Hz,1H),2.29(s,6H), 2.07(s,3H),1.95-1.73(m,5H),1.60-1.55(m,1H),1.49(s,3H),1.42-1.34(m,1H),1.33(s,3H),1. 29(d,J=6.9Hz,3H),1.27-1.22(m,7H),1.22-1.17(m,1H),0.95(d,J=7.0Hz,3H),0.92-0.85(m,6H).

[0073] Synthesis of intermediate compound 7

[0074]

[0075] Intermediate compound 5 (0.560 g, 0.76 mmol) was dissolved in acetic acid (6 mL, 10 times the mass of intermediate compound 5) and stirred until homogeneous. Platinum dioxide (0.056 g, 0.1 times the mass of intermediate compound 5) was then added. Following the general synthetic method of this preparation example, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.6 / 0.1) to give a white, fluffy solid intermediate compound 7 (0.240 g, 0.32 mmol, 42.1%). HRMS (ESI) (M+H) + m / z 744.4645, calcd for C 37 H 66 N3O 12 744.4641. 1H NMR (CDCl3, 400MHz) δ: 5.04 (dd, J1 = 8.4Hz, J2 = 3.8Hz, 1H), 4.96 (s, 1H), 4.76 (dd, J1 = 10.5Hz, J2 = 7.4Hz, 1H), 4.35 (d, J = 7 .4Hz,1H),4.08(s,1H),3.80(s,3H),3.76-3.55(m,3H),3.48-3.39(m,1H),3.10(br,1H),2.98-2.84(m,6H),2.75-2.65( m,1H),2.51(q,J=6.8Hz,1H),2.27(s,6H),2.13-2.07(m,1H),2.05(s,3H),1.97-1.67(m,4H),1.58-1.52(m,1H),1.50(s ,3H),1.42-1.34(m,2H),1.31(d,J=7.1Hz,3H),1.29(s,3H),1.26-1.20(m,7H),0.97(d,J=7.1Hz,3H),0.95-0.89(m,6H).

[0076] Preparation Example 3.3: General Synthetic Method for Intermediate Compounds 8-13

[0077] Intermediate compounds 6 or 7 (1 eq) were dissolved in 3-butyn-1-ol / 4-pentyn-1-ol / 5-hexyn-1-ol / 6-heptyyn-1-ol / 7-octyyn-1-ol (100 eq), and formic acid (4.5 eq) and sodium nitrite (6 eq) were added in portions at -15 °C. The reaction was carried out for approximately 5-6 days, and the reaction progress was monitored by TLC. After the reaction was completed, the pH was adjusted to 9-10 with 2M NaOH solution. The organic phase was washed once with distilled water and once with saturated NaCl solution, and purified by rotary cyclohexane column chromatography to obtain intermediate compounds 8-13.

[0078] Synthesis of intermediate compound 8

[0079]

[0080] Using compound 6 (0.773 g, 1.04 mmol) and 3-butyn-1-ol (8 mL, 104.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give compound 8 (0.120 g, 0.15 mmol, 14.4%) as a white, fluffy solid.

[0081] Synthesis of intermediate compound 9

[0082]

[0083] Using intermediate compound 6 (0.576 g, 0.77 mmol) and 4-pentyn-1-ol (7 mL, 77.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give a white, fluffy solid intermediate compound 9 (0.128 g, 0.16 mmol, 20.8%). HRMS(ESI)(M+H) + m / z 811.49497, calcd for C 42 H 71 N2O 13 811.4951. 1 H NMR (CDCl3, 400MHz) δ: 5.16 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H), 4.77 (dd, J1 = 10.5Hz, J2 = 7.4Hz, 1H), 4.44 (d, J = 7.5Hz, 1H), 3.84 (s, 3H), 3.83-3.78(m,1H),3.72-3.64(m,3H),3.58-3.50(m,4H),3.50-3.44(m,1 H),3.19(d,J=10.5Hz,1H),2.98(s,3H),2.84-2.68(m,2H),2.47(q,J=6. 9Hz,1H),2.33-2.26(m,8H),2.07(s,3H),1.97(t,J=2.7Hz,1H),1.96-1. 85(m,4H),1.85-1.72(m,3H),1.59-1.53(m,1H),1.49(s,3H),1.42-1.35 (m,1H),1.33(s,3H),1.31-1.27(m,4H),1.25(d,J=1.7Hz,3H),1.24(d,J =2.4Hz,3H),1.22-1.16(m,1H,0.95(d,J=7.1Hz,3H),0.92-0.85(m,6H).

[0084] Synthesis of intermediate compound 10

[0085]

[0086] Using intermediate compound 6 (1.657 g, 2.23 mmol) and 5-hexyn-1-ol (24 mL, 223.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give a white, fluffy solid intermediate compound 10 (0.153 g, 0.19 mmol, 8.5%).

[0087] Synthesis of intermediate compound 11

[0088]

[0089] Using intermediate compound 6 (1.137 g, 1.53 mmol) and 6-heptyne-1-ol (19 mL, 153.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give a white, fluffy solid intermediate compound 11 (0.124 g, 0.15 mmol, 9.8%).

[0090] Synthesis of intermediate compound 12

[0091]

[0092] Using intermediate compound 6 (1.041 g, 1.40 mmol) and 7-octyne-1-ol (19 mL, 140.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give intermediate compound 12 (0.117 g, 0.14 mmol, 10.0%), a white, fluffy solid.

[0093] Synthesis of intermediate compound 13

[0094]

[0095] Using intermediate compound 7 (1.520 g, 2.04 mmol) and 5-hexyn-1-ol (22 mL, 204.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.05 / 0.05) to give a white, fluffy solid intermediate compound 13 (0.167 g, 0.20 mmol, 9.9%). HRMS(ESI)(M+H) + m / z 825.5100, calcd for C 43H 73 N2O 13 825.5107. 1 H NMR (CDCl3, 400MHz) δ: 5.02 (dd, J1 = 8.1Hz, J2 = 3.7Hz, 1H), 4.96 (s, 1H), 4.75 (dd, J1 = 10.6Hz, J2 = 7.5Hz, 1H), 4.37 (d, J = 7.4Hz, 1H), 4.07 (s, 1H), 3.80(s,3H),3.77-3.48(m,5H),3.48-3.40(m,3H),3.10(m,1H),2.93(s, 3H),2.92(s,1H),2.75-2.66(m,1H),2.52(q,J=6.9Hz,1H),2.27(s,6H), 2.23-2.19(td,J1=7.2Hz,J2=2.8Hz,2H),2.11-2.08(m,1H),2.04(s,3H) ,1.93(t,J=2.7Hz,1H),1.91-1.85(m,2H),1.75-1.64(m,5H),1.762-1.5 3(m,2H),1.49(s,3H),1.40-1.34(m,2H),1.30-1.29(d,,J=4.2Hz,3H),1 .27(s,3H),1.24-1.21(m,7H),0.97(d,J=7.0Hz,3H),0.94-0.90(m,6H).

[0096] Preparation Example 3.4: Preparation of Intermediate Compound 14

[0097]

[0098] The obtained intermediate compound 6 (1.679 g, 2.26 mmol) was placed in a dry 100 mL round-bottom flask, and 5-hexen-1-ol (27 mL, 226 mmol) was added. The mixture was stirred thoroughly in an ice-salt bath, and formic acid (0.38 mL, 10.17 mmol) was added. Sodium nitrite (0.936 g, 13.56 mmol) was added in portions. The reaction was allowed to proceed for approximately 5-6 days, and the reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 9-10 with 2 M NaOH solution. The organic phase was washed once with distilled water and once with saturated NaCl solution, and the solvent was removed under reduced pressure. The mixture was then purified by column chromatography. The column chromatography conditions were: 100-200 mesh silica gel, and the mobile phase was dichloromethane / ethanol = 10 / 0.1. This yielded a white, fluffy solid intermediate compound 14 (0.180 g, 0.22 mmol, 9.7%).

[0099] Preparation Example 3.5: General Synthetic Method for Intermediate Compounds 15-19

[0100] Intermediate compounds 6 or 7 (1 eq) were dissolved in 1,2-ethylene glycol / 1,3-propanediol / 1,4-butanediol / 1,5-pentanediol (100 eq), and formic acid (4.5 eq) and sodium nitrite (6 eq) were added in portions at -15°C. The reaction was allowed to proceed for approximately 5-6 days, and the reaction was monitored by TLC. After the reaction was complete, the pH was adjusted to 9-10 with 2M NaOH solution. The organic phase was washed once with distilled water and once with saturated NaCl solution, and purified by rotary cyclohexane column chromatography to obtain intermediate compounds 15-19.

[0101] Synthesis of intermediate compound 15

[0102]

[0103] Using intermediate compound 6 (0.804 g, 1.08 mmol) and 1,2-ethylene glycol (6 mL, 108.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.10 / 0.05) to give a white, fluffy solid intermediate compound 15 (0.386 g, 0.49 mmol, 45.4%). HRMS(ESI)(M+H) + m / z789.4741, calcd for C 39 H 69 N2O 14 789.4743. 1 H NMR (CDCl3, 400MHz) δ: 5.15 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H), 4.77 (dd, J1 = 10.5Hz, J2 = 7.5Hz, 1H), 4.44 (d, J = 7.5 Hz,1H),3.88-3.79(m,4H),3.79-3.56(m,9H),3.52-3.43(m,1H),3.20(d,J=10.4Hz,1H),2.98(s,3H),2.85-2.69(m,2H),2. 47(q,J=6.7Hz,1H),2.29(s,6H),2.07(s,3H),2.01-1.86(m,4H),1.79-1.72(m,1H),1.62-1.52(m,1H),1.49(s,3H),1.42-1 .35(m,1H),1.33(s,3H),1.31-1.27(m,4H),1.27-1.22(m,6H),1.22-1.15(m,1H),0.95(d,J=7.1Hz,3H),0.92-0.84(m,6H).

[0104] Synthesis of intermediate compound 16

[0105]

[0106] Using intermediate compound 6 (0.968 g, 1.30 mmol) and 1,3-propanediol (9 mL, 130.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.10 / 0.05) to give intermediate compound 16 (0.390 g, 0.49 mmol, 37.7%) as a white, fluffy solid.

[0107] Synthesis of intermediate compound 17

[0108]

[0109] Using intermediate compound 6 (1.414 g, 1.90 mmol) and 1,4-butanediol (17 mL, 190.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.10 / 0.05) to give intermediate compound 17 (0.673 g, 0.82 mmol, 43.2%), a white, fluffy solid.

[0110] Synthesis of intermediate compound 18

[0111]

[0112] Using intermediate compound 6 (1.401 g, 1.88 mmol) and 1,5-pentanediol (20 mL, 188.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.10 / 0.05) to give intermediate compound 18 (0.448 g, 0.54 mmol, 28.7%), a white, fluffy solid.

[0113] Synthesis of intermediate compound 19

[0114]

[0115] Using intermediate compound 7 (3.05 g, 4.10 mmol) and 1,4-butanediol (39 mL, 410.00 mmol) as starting materials, the product was purified by column chromatography (100-200 mesh silica gel, mobile phase: dichloromethane / ethanol / ammonia = 10 / 0.10 / 0.05) to give intermediate compound 19 (0.610 g, 0.75 mmol, 18.2%), a white, fluffy solid.

[0116] Preparation of target compounds 20-25

[0117]

[0118] General synthetic method for target compounds 20-25

[0119] The iodoquinolone derivative (1.2 eq) was placed in a pressure flask and dissolved in 5 mL of acetonitrile and 5 mL of triethylamine. Cuprous iodide (0.1 eq) was then added, followed by argon purging. The flask was sealed and stirred at room temperature for 20 min. Then, intermediate compound 8-13 (1 eq) and bis(triphenylphosphine)palladium(II) dichloride (0.05 eq) were added, followed by argon purging to ensure the pressure flask was completely filled with argon. The flask was sealed and reacted in an oil bath at 45 °C. The reaction was monitored by TLC for approximately 12 h. After the reaction, the organic phase was washed with saturated NaCl solution, and the solvent was removed under reduced pressure. The crude product was purified by column chromatography. The obtained compound was dissolved in 10 mL of methanol and refluxed at 65 °C. The reaction was monitored by TLC for approximately 2 h. After the reaction, the reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography to obtain the target compound 20-25.

[0120] The quinolone derivatives used in the synthesis are shown as a-Ⅰ:

[0121] aI

[0122]

[0123] Synthesis of target compound 20

[0124]

[0125] According to the general synthetic method of this embodiment, intermediate compound 8 (0.120 g, 0.15 mmol) and compound aI (0.059 g, 0.18 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 20 (20 mg, 0.02 mmol, 13.3%). HRMS(ESI)(M+H) + m / z956.51202, calcd for C 50 H74 N3O 15 956.5114. 1 H NMR (CDCl3, 400MHz) δ: 8.77 (s, 1H), 8.47 (d, J = 8.3Hz, 1H), 7.63 (s, 1H), 7.58 (d, J = 8.3Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H),4.37(d,J=7.3Hz,1H),4.02(s,3H),3.88-3.80(m,4H),3.76(d,J=3.4Hz,1H),3.73-3.58(m,6H),3.53-3.43(m,1H),3.27-3.18(m,2H), 2.98(s,3H),2.86-2.69(m,3H),2.56-2.43(m,2H),2.29(s,6H),2.00 -1.84(m,4H),1.71-1.64(m,1H),1.62-1.52(m,1H),1.49(s,3H),1.47 -1.39(m,1H),1.38-1.31(m,4H),1.31-1.26(m,4H),1.26-1.21(m,6H),1.02(d,J=7.3Hz,3H),0.96(d,J=7.0Hz,3H),0.86(t,J=7.3Hz,3H). 13 C NMR(CDCl3,175MHz)δ:178.09,175.48,166.82,164.63,154.76,149.50,140.11,129.91,129.39,1 27.10,125.25,119.25,109.10,101.65,92.60,85.19,85.06,83.12,80.32,79.79,78.52,75.27,7 1.15,70.63,69.28,68.74,67.99,65.59,61.42,49.87,44.88,42.06,40.38,37.29,36.98,32.71,31.00,29.72,28.77,25.64,22.17,21.32,20.95,19.50,18.83,15.58,15.43,12.96,10.19,8.63.

[0126] Synthesis of target compound 21

[0127]

[0128] According to the general synthetic method of this embodiment, intermediate compound 9 (0.128 g, 0.16 mmol) and compound aI (0.063 g, 0.19 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding target compound 21 (22 mg, 0.02 mmol, 12.5%). HRMS(ESI)(M+H) + m / z970.52824, calcd for C 51 H 76 N3O 15 970.5271. 1 H NMR (CDCl3, 400MHz) δ: 8.77 (s, 1H), 8.46 (d, J = 8.3Hz, 1H), 7.62 (s, 1H), 7.57 (d, J = 8.4Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H), 4.38(d,J=7.3Hz,1H),4.02(s,3H),3.87-3.80(m,4H),3.76(d,J=3.5Hz ,1H),3.74-3.63(m,2H),3.61-3.53(m,4H),3.52-3.46(m,1H),3.27-3.1 7(m,2H),2.99(s,3H),2.86-2.77(m,1H),2.63-2.44(m,4H),2.30(s,6H ),2.00-1.82(m,6H),1.73-1.65(m,1H),1.62-1.52(m,1H),1.49(s,3H), 1.46-1.40(m,1H),1.39-1.31(m,4H),1.31-1.26(m,4H),1.26-1.19(m, 6H), 1.02 (d, J = 7.4Hz, 3H), 0.95 (d, J = 7.0Hz, 3H), 0.86 (t, J = 7.3Hz, 3H). 13C NMR(CDCl3,100MHz)δ:178.11,175.51,166.82,164.68,154.75,149.47,140.17,130.25,129.3 8,127.07,125.14,119.09,109.10,101.69,95.50,85.09,83.11,79.84,78.55,75.27,71.30,7 0.68,69.36,67.79,65.61,61.38,49.86,44.92,42.02,40.37,37.30,37.02,32.73,31.10,29.70,28.82,28.66,25.65,22.18,21.29,19.48,18.82,16.48,15.55,15.39,12.93,10.18,8.61.

[0129] Synthesis of target compound 22

[0130]

[0131] According to the general synthetic method of this embodiment, intermediate compound 10 (0.153 g, 0.19 mmol) and compound aI (0.073 g, 0.22 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding target compound 22 (21 mg, 0.02 mmol, 10.5%). HRMS(ESI)(M+H) + m / z984.54325, calcd for C 52 H 78 N3O 15 984.5427. 1H NMR (CDCl3, 400MHz) δ: 8.67 (s, 1H), 8.37 (d, J = 8.3Hz, 1H), 7.52 (s, 1H), 7.48 (d, J = 8.4Hz, 1H), 5.05 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.77 (s, 1H),4.29(d,J=7.2Hz,1H),3.93(s,3H),3.82-3.70(m,4H),3.67(d,J=3.4Hz,1H),3.65-3.51(m,2H),3.49-3.32(m,5H),3.18-3.06(m,2H), 2.90(s,3H),2.78-2.68(m,1H),2.48-2.35(m,4H),2.21(s,6H),1.90 -1.73(m,4H),1.72-1.54(m,5H),1.53-1.43(m,1H),1.40(s,3H),1.38 -1.31(m,1H),1.29-1.23(m,4H),1.22-1.17(m,4H),1.17-1.12(m,6H),0.93(d,J=7.3Hz,3H),0.86(d,J=6.9Hz,3H),0.78(t,J=7.3Hz,3H). 13 C NMR(CDCl3,100MHz)δ:178.11,175.51,166.81,164.72,154.73,149.48,140.18,130.33,129.40 ,127.04,125.12,119.05,109.09,101.69,95.85,85.18,85.09,83.10,79.83,78.57,75.25,71.3 5,70.71,70.42,69.25,67.78,65.59,61.36,49.86,44.92,42.00,40.36,37.30,37.04,32.74,31.12,29.07,28.84,25.66,25.28,22.19,21.28,19.46,18.82,15.54,15.38,12.91,10.17,8.60.

[0132] Synthesis of target compound 23

[0133]

[0134] According to the general synthetic method of this embodiment, intermediate compound 11 (0.124 g, 0.15 mmol) and compound aI (0.059 g, 0.18 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding target compound 23 (21 mg, 0.02 mmol, 13.3%). HRMS(ESI)(M+H) + m / z998.5561, calcd for C 53 H 80 N3O 15 998.5584. 1 H NMR (CDCl3, 400MHz) δ: 8.76 (s, 1H), 8.45 (d, J = 8.3Hz, 1H), 7.60 (s, 1H), 7.57 (d, J = 8.4Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H),4.37(d,J=7.2Hz,1H),4.02(s,3H),3.86-3.78(m,4H),3.76(d,J=3.3Hz,1H),3.73-3.61(m,2H),3.56-3.38(m,5H),3.27-3.16(m,2H), 2.99(s,3H),2.87-2.75(m,1H),2.57-2.41(m,4H),2.30(s,6H),1.99 -1.81(m,4H),1.75-1.60(m,5H),1.60-1.51(m,3H),1.49(s,3H),1.46 -1.39(m,1H),1.38-1.30(m,4H),1.30-1.26(m,4H),1.26-1.18(m,6H),1.01(d,J=7.4Hz,3H),0.95(d,J=7.0Hz,3H),0.87(t,J=7.3Hz,3H). 13C NMR(CDCl3,175MHz)δ:178.10,175.51,166.88,164.69,154.78,149.49,140.15,130.39,129.43,12 7.00,125.04,119.09,109.00,101.64,96.13,85.13,85.10,83.13,79.77,79.60,78.51,75.21,71.3 6,70.88,70.63,69.26,67.73,65.56,61.40,49.87,44.88,42.06,40.37,37.26,36.97,32.70,31.12,29.32,28.70,28.33,25.70,25.65,22.16,21.31,19.50,18.82,15.57,15.39,12.95,10.20,8.61.

[0135] Synthesis of target compound 24

[0136]

[0137] According to the general synthetic method of this embodiment, intermediate compound 12 (0.117 g, 0.14 mmol) and compound aI (0.055 g, 0.17 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 24 (22 mg, 0.02 mmol, 14.3%). HRMS(ESI)(M+H) + m / z1012.5767, calcd for C 54 H 82 N3O 15 1012.5740. 1H NMR (CDCl3, 400MHz) δ: 8.77 (s, 1H), 8.45 (d, J = 8.4Hz, 1H), 7.60 (s, 1H), 7.56 (d, J = 8.5Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.8 6(s,1H),4.37(d,J=7.1Hz,1H),4.03(s,3H),3.88-3.79(m,4H),3.76(d,J=3.3Hz,1H),3.73-3.61(m,2H),3.56-3.37(m,5H),3.29-3.1 4(m,2H),2.99(s,3H),2.86-2.77(m,1H),2.55-2.40(m,4H),2.30(s,6H),1.97-1.83(m,4H),1.76-1.52(m,8H),1.49(s,3H),1.46-1.3 9(m,3H),1.37-1.31(m,4H),1.30-1.26(m,4H),1.25-1.20(m,6H),1.02(d,J=7.3Hz,3H),0.95(d,J=6.9Hz,3H),0.87(t,J=7.3Hz,3H). 13 C NMR(CDCl3,175MHz)δ:178.10,175.51,166.82,164.66,154.74,149.46,140.16,130.42,129.41,127.0 1,125.05,119.04,109.04,101.66,96.26,85.15,85.06,83.12,79.80,79.55,78.52,75.21,71.39,71.0 1,70.65,69.24,67.69,65.58,61.37,49.85,44.89,42.02,40.36,37.27,36.99,32.70,31.12,29.69,28.90,28.73,28.38,25.81,25.63,22.17,21.28,19.53,19.47,18.82,15.56,15.37,12.93,10.18,8.60.

[0138] Synthesis of target compound 25

[0139]

[0140] According to the general synthetic method of this embodiment, intermediate compound 13 (0.167 g, 0.20 mmol) and compound aI (0.080 g, 0.24 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 25 (33 mg, 0.03 mmol, 16.6%). HRMS(ESI)(M+H) + m / z984.5416, calcd for C 52 H 78 N3O 15 984.5427. 1 H NMR (MeOD, 400MHz) δ: 8.73 (s, 1H), 8.25 (d, J = 8.2Hz, 1H), 7.68 (s, 1H), 7.39 (s, 1H), 4.94 (s, 1H), 4.82-4 .78(m,1H),4.14(m,2H),3.92(s,3H),3.67(s,3H),3.64-3.38(m,8H),3.17-3.14(m,1H),3.07-3.03(m, 1H),2.89-2.86(m,4H),2.7-2.52(m,2H),2.48-2.39(m,2H),2.29(s,6H),2.03-1.92(m,1H),1.84-1.72 (m,3H),1.71-1.59(m,5H),1.53-1.30(m,6H),1.24-1.15(m,7H),1.14-1.07(m,6H),0.88-0.79(m,6H). 13 C NMR(MeOD,100MHz)δ:154.77,149.40,140.16,130.42,129.46,127.07,125.11,119.05,109.09,85.51,83.00,78.19,77.20,77.0 2,76.84,75.86,70.44,70.35,69.62,67.53,65.77,61.35,42.05,40.35,37.96,35.15,30.78,29.05,28.53,26.01,25.25,21.25.

[0141] Synthesis of target compound 26

[0142]

[0143] Intermediate compound 14 (0.110 g, 0.13 mmol) was placed in a pressure vessel and dissolved in 5 mL of DMF. Iodoquinolone derivative (0.051 g, 0.16 mmol), sodium acetate (0.039 g, 0.48 mmol), trans-di-(M)-bis[2-(di-o-tolylphosphine)benzyl]acetate dipalladium(II) (0.012 g, 0.01 mmol), and tetrabutylammonium bromide (0.838 g, 2.60 mmol) were then added. Argon purging was performed to ensure the pressure vessel was completely filled with argon. The vessel was sealed and reacted in a 60°C oil bath for 1 h. The temperature was then increased to 100°C and the reaction continued for approximately 72 h. The reaction progress was monitored by TLC. After the reaction was complete, the organic phase was washed with a semi-saturated NaCl solution, the solvent was removed under reduced pressure, and the mixture was purified by column chromatography. The column chromatography conditions were: 100-200 mesh silica gel, mobile phase: dichloromethane / methanol / ammonia = 10 / 0.1 / 0.5. The obtained compound was dissolved in 10 mL of methanol and refluxed at 65 °C. The reaction was carried out for approximately 2 h, and the reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed under reduced pressure and purified by column chromatography. The column chromatography conditions were: 100-200 mesh silica gel, mobile phase: dichloromethane / methanol / ammonia = 10 / 0.1 / 0.5, thus yielding the target compound 26 (25 mg, 0.03 mmol, 23.1%). HRMS (ESI) (M+H) + m / z 986.5584, calcd for C 52 H 80 N3O 15 986.5584. 1H NMR (CDCl3, 400MHz) δ: 8.65 (s, 1H), 8.36 (d, J = 8.5Hz, 1H), 7.55 (d, J = 8.5Hz, 1H), 7.33 (s, 1H), 6.54-6.27 (m, 2H), 5.05 (dd, J1 = 10.7Hz, J2 = 2. 4Hz,1H),4.77(s,1H),4.28(d,J=7.2Hz,1H),3.95(s,3H),3.79-3.69(m,4H),3.67(d,J=3.5Hz,1H),3.65-3.53(m,2H),3.49-3.30(m,5H),3. 16-3.10(m,2H),2.90(s,3H),2.79-2.68(m,1H),2.45-2.35(m,2H),2. 29-2.14(m,8H),1.90-1.73(m,4H),1.63-1.43(m,6H),1.40(s,3H),1. 38-1.31(m,1H),1.28-1.22(m,4H),1.22-1.17(m,4H),1.17-1.09(m,6 H), 0.93 (d, J = 7.4Hz, 3H), 0.86 (d, J = 7.0Hz, 3H), 0.78 (t, J = 7.4Hz, 3H). 13 CNMR(CDCl3,175MHz)δ:178.19,175.54,167.13,164.65,154.77,149.16,143.87,140.77,136.52, 128.79,127.23,124.77,123.69,113.60,108.77,101.65,85.13,85.08,83.12,79.76,78.51,75.24 ,71.34,70.82,70.64,69.28,67.77,65.58,61.41,49.88,44.89,42.03,40.38,37.27,36.98,33.07,32.71,31.14,29.41,28.68,25.65,22.17,21.32,19.50,18.83,15.58,15.41,12.95,10.20,8.62.

[0144] Preparation Example 3.6: General Synthetic Method for Intermediate Compounds 27-31

[0145]

[0146] Intermediate compound 15-19 (1 eq) was dissolved in dry DCM (10 mL), and triethylamine (1.5 eq), DMAP (1.5 eq), and p-toluenesulfonyl chloride (1.5 eq) were added. The reaction was carried out at room temperature for about 2-3 h, and the reaction progress was monitored by TLC. After the reaction was completed, the organic phase was washed five times with saturated NH4Cl solution for about 20 min each time, and then washed once with distilled water and once with saturated NaCl solution. The organic phase was purified by rotary evaporation column chromatography to obtain intermediate compound 27-31.

[0147] Synthesis of intermediate compound 27

[0148]

[0149] According to the general synthesis method of this embodiment, intermediate compound 15 (0.386 g, 0.49 mmol) was used as the starting material for the reaction. The column chromatography conditions were 100-200 mesh silica gel and the mobile phase was dichloromethane / ethanol / ammonia water = 10 / 0.1 / 0.05 to obtain intermediate compound 27 (0.120 g, 0.14 mmol, 28.6%).

[0150] Synthesis of intermediate compound 28

[0151]

[0152] According to the general synthesis method of this embodiment, intermediate compound 16 (0.390 g, 0.49 mmol) was used as the starting material for the reaction. The column chromatography conditions were 100-200 mesh silica gel and the mobile phase was ethyl acetate / petroleum ether = 1 / 1 to obtain intermediate compound 28 (0.171 g, 0.18 mmol, 36.7%).

[0153] Synthesis of intermediate compound 29

[0154]

[0155] According to the general synthetic method of this embodiment, intermediate compound 17 (0.673 g, 0.82 mmol) was used as the starting material. Column chromatography conditions were 100-200 mesh silica gel, and the mobile phase was ethyl acetate / petroleum ether = 1 / 1, yielding intermediate compound 29 (0.313 g, 0.32 mmol, 39.0%). HRMS(ESI)(M+H) + m / z 971.5162, calcd for C 48 H 79 N2O 16 S971.5145. 1H NMR (CDCl3, 400MHz) δ: 7.78 (d, J = 8.4Hz, 2H), 7.35 (d, J = 8.0Hz, 2H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.82 (s, 1H), 4.74 (dd, J1 = 10. 6Hz,J2=7.4Hz,1H),4.40(d,J=7.5Hz,1H),4.08-4.01(m,2H),3.81(s,3H),3.80-3.72(m,1H),3.67(d,J=3.5Hz,1H),3.65-3.56(m,2H), 3.53-3.35(m,5H),3.15(d,J=10.5Hz,1H),2.95(s,3H),2.82-2.63(m,2H),2.49-2.40(m,4H),2.27(s,6H),2.04(s,3H),1.96-1.80(m,4 H),1.77-1.67(m,3H),1.65-1.50(m,3H),1.47(s,3H),1.33-1.28(m,4H),1.28-1.13(m,11H),0.93(d,J=7.0Hz,3H),0.90-0.82(m,6H).

[0156] Synthesis of intermediate compound 30

[0157]

[0158] According to the general synthesis method of this embodiment, intermediate compound 18 (0.448 g, 0.54 mmol) was used as the starting material for the reaction. The column chromatography conditions were 100-200 mesh silica gel and the mobile phase was ethyl acetate / petroleum ether = 1 / 1 to obtain intermediate compound 30 (0.342 g, 0.35 mmol, 64.8%).

[0159] Synthesis of intermediate compound 31

[0160]

[0161] According to the general synthesis method of this embodiment, intermediate compound 19 (0.610 g, 0.75 mmol) was used as the starting material for the reaction. The column chromatography conditions were 100-200 mesh silica gel and the mobile phase was ethyl acetate / petroleum ether = 1 / 1 to obtain intermediate compound 31 (0.221 g, 0.23 mmol, 30.5%).

[0162] Preparation of target compounds 32x-36x

[0163]

[0164] Where x is the number a, b or c corresponding to the Ar group selected in the target compound. For example, when Ar in the target compound 32x is selected from group a, the target compound is numbered 32a.

[0165] General synthetic method for target compounds 32x-36x

[0166] Compounds 27-31 (1 eq) were placed in a dry 100 mL round-bottom flask and dissolved in 10 mL of acetonitrile with stirring. Ciprofloxacin / enoxacin / norfloxacin (Ar = a, b, c) (3 eq) were then added, and the reaction was carried out in an oil bath at 70 °C. The reaction was allowed to proceed for approximately 7-8 days, and the reaction progress was monitored by TLC. After the reaction was complete, the solvent was removed under reduced pressure, and the crude product was purified by column chromatography. The obtained compound was dissolved in 10 mL of methanol and refluxed at 65 °C. The reaction was allowed to proceed for approximately 2 hours, and the reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography to obtain the target compound 32x-36x.

[0167] Synthesis of target compound 32a

[0168]

[0169] According to the general synthetic method of this embodiment, intermediate compound 27 (0.120 g, 0.14 mmol) and ciprofloxacin (Ar=a) (0.139 g, 0.42 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 32a (28 mg, 0.03 mmol, 21.4%) was obtained. HRMS (ESI) (M+H) + m / z1060.5873, calcd for C 54 H 83 FN5O 15 1060.5864. 1H NMR (CDCl3, 400MHz) δ: 8.79 (s, 1H), 8.04 (d, J = 13.0Hz, 1H), 7.38 (d, J = 7.0Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H), 4.38 (d, J = 7.2 Hz,1H),3.90-3.79(m,4H),3.77(d,J=3.4Hz,1H),3.74-3.53(m,7H),3.52 -3.44(m,1H),3.43-3.33(m,4H),3.28-3.19(m,2H),3.00(s,3H),2.88-2. 80(m,1H),2.80-2.74(m,4H),2.73-2.63(m,2H),2.60-2.45(m,2H),2.33( s,6H),2.02-1.83(m,4H),1.77-1.68(m,1H),1.63-1.53(m,1H),1.50(s,3 H),1.47-1.38(m,3H),1.38-1.32(m,5H),1.31-1.27(m,4H),1.27-1.20(m ,7H),1.03(d,J=7.4Hz,3H),0.96(d,J=7.0Hz,3H),0.88(t,J=7.4Hz,3H). 13 C NMR(CDCl3,175MHz)δ:177.16,175.50,167.04,164.59,154.72,152.98,147.45,145.93,145.87,139. 11,119.91,119.87,112.54,112.41,108.21,104.75,101.66,85.19,85.05,83.11,79.82,78.54,75.29 ,71.31,70.66,69.27,68.73,68.17,65.62,61.41,57.79,53.31,49.88,49.76,44.89,40.40,37.30,37.01,35.28,32.73,31.03,25.64,22.19,21.31,19.49,18.83,15.57,15.41,12.95,10.20,8.62,8.25.

[0170] Synthesis of target compound 32b

[0171]

[0172] According to the general synthetic method of this embodiment, intermediate compound 27 (0.171 g, 0.18 mmol) and enoxacin (Ar=b) (0.173 g, 0.54 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 32b (35 mg, 0.03 mmol, 16.7%) was obtained. HRMS (ESI) (M+H) + m / z1049.5819, calcd for C 52 H 82 FN6O 15 1049.5817. 1 H NMR (CDCl3, 400MHz) δ: 8.69 (s, 1H), 8.08 (d, J = 13.4Hz, 1H), 5.13 (dd, J1 = 10.9Hz, J2 = 2.6Hz, 1H), 4.84 (s, 1H), 4.50-4.23 (m, 3H), 3.96-3.86 (m ,4H),3.85-3.77(m,4H),3.75(d,J=3.4Hz,1H),3.72-3.64(m,2H),3.62 -3.51(m,4H),3.51-3.41(m,1H),3.25-3.14(m,2H),2.98(s,3H),2.86- 2.76(m,1H),2.72-2.58(m,6H),2.53-2.43(m,2H),2.30(s,6H),2.00- 1.82(m,4H),1.74-1.64(m,1H),1.61-1.54(m,1H),1.53-1.46(m,6H),1 .45-1.40(m,1H),1.36-1.30(m,4H),1.29-1.25(m,4H),1.25-1.19(m,6 H), 1.01 (d, J = 7.4Hz, 3H), 0.94 (d, J = 7.0Hz, 3H), 0.86 (t, J = 7.3Hz, 3H). 13C NMR(CDCl3,175MHz)δ:177.04,175.46,166.96,164.66,164.62,154.72,150.42,150.37,148.08,146.35, 145.07,120.23,120.10,113.73,113.71,109.27,101.65,85.16,85.04,83.12,79.76,78.53,75.25,75.21 ,71.25,70.63,69.26,68.65,68.19,65.61,61.39,57.75,53.47,49.86,47.76,46.97,46.92,44.87,40.37,37.29,36.99,32.71,31.00,25.63,22.18,21.30,19.48,18.82,15.56,15.38,14.97,12.94,10.18,8.59.

[0173] Synthesis of target compound 33a

[0174]

[0175] According to the general synthetic method of this embodiment, intermediate compound 28 (0.171 g, 0.18 mmol) and ciprofloxacin (Ar=a) (0.179 g, 0.54 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 33a (38 mg, 0.04 mmol, 22.2%) was obtained. HRMS (ESI) (M+H) + m / z1074.60254, calcd for C 55 H 85 FN5O 15 1074.6021. 1H NMR (CDCl3, 400MHz) δ: 8.76 (s, 1H), 8.00 (d, J = 13.1Hz, 1H), 7.37 (d, J = 7.0Hz, 1H), 5.14 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.85 (s, 1H), 4.38 (d, J =7.2Hz,1H),3.87-3.78(m,4H),3.76(d,J=3.4Hz,1H),3.73-3.61(m,2H ),3.60-3.42(m,6H),3.41-3.32(m,4H),3.26-3.18(m,2H),2.99(s,3H) ,2.88-2.77(m,1H),2.74-2.58(m,4H),2.57-2.42(m,4H),2.30(s,6H), 2.00-1.74(m,6H),1.71-1.64(m,1H),1.61-1.52(m,1H),1.49(s,3H),1 .47-1.38(m,3H),1.38-1.32(m,5H),1.31-1.27(m,4H),1.26-1.18(m,7 H), 1.02 (d, J = 7.4Hz, 3H), 0.95 (d, J = 7.0Hz, 3H), 0.87 (t, J = 7.3Hz, 3H). 13 C NMR(CDCl3,175MHz)δ:177.11,175.50,167.03,164.63,154.74,152.97,147.39,145.94,145.88,139.11 ,119.79,119.74,112.45,112.32,108.12,104.74,101.67,85.15,85.06,83.13,79.78,78.53,75.24,71 .34,70.67,69.28,69.21,67.78,65.61,61.39,55.42,52.87,49.87,44.90,40.38,37.29,37.00,35.29,32.71,31.11,28.74,27.19,25.64,22.18,21.32,19.48,18.83,15.57,15.40,12.94,10.19,8.60,8.24.

[0176] Synthesis of target compound 33b

[0177]

[0178] According to the general synthetic method of this embodiment, intermediate compound 28 (0.100 g, 0.10 mmol) and enoxacin (Ar=b) (0.096 g, 0.30 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 33b (35 mg, 0.03 mmol, 30.0%) was obtained. HRMS (ESI) (M+H) + m / z1063.5991, calcd for C 53 H 84 FN6O 15 1063.5973. 1 H NMR (CDCl3, 400MHz) δ: 8.69 (s, 1H), 8.10 (d, J = 13.4Hz, 1H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.84 (s, 1H), 4.46-4.31 (m, 3H), 3.94-3. 85(m,4H),3.85-3.77(m,4H),3.75(d,J=3.4Hz,1H),3.72-3.60(m,2H),3.57-3.41(m,5H),3.27-3.16(m,2H),2.98(s,3H),2.86-2.76(m,1 H),2.65-2.54(m,4H),2.53-2.41(m,4H),2.29(s,6H),1.99-1.74(m,6H),1.70-1.63(m,1H),1.61-1.53(m,1H),1.53-1.46(m,6H),1.45- 1.40(m,1H),1.35-1.30(m,4H),1.29-1.25(m,4H),1.25-1.21(m,6H),1.01(d,J=7.4Hz,3H),0.94(d,J=7.0Hz,3H),0.86(t,J=7.4Hz,3H). 13C NMR(CDCl3,175MHz)δ:177.09,175.50,166.99,164.61,154.74,150.46,150.41,148.10,146.36,145.10, 120.30,120.18,113.79,109.33,101.67,85.16,85.05,83.13,79.79,78.54,75.27,71.30,70.68,70.68, 69.29,69.13,67.80,65.61,61.41,55.35,53.05,49.87,47.76,47.06,47.01,44.91,40.38,37.29,37.01,32.72,31.11,28.75,27.14,25.64,22.19,21.31,19.48,18.84,15.57,15.39,14.98,12.95,10.19,8.60.

[0179] Synthesis of target compound 34a

[0180]

[0181] According to the general synthetic method of this embodiment, intermediate compound 29 (0.166 g, 0.17 mmol) and ciprofloxacin (Ar=a) (0.169 g, 0.51 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 34a (38 mg, 0.04 mmol, 23.5%) was obtained. HRMS (ESI) (M+H) + m / z1088.6191, calcd for C 56 H 87 FN5O 15 1088.6177. 1H NMR (CDCl3, 400MHz) δ: 8.77 (s, 1H), 8.01 (d, J = 13.1Hz, 1H), 7.36 (d, J = 7.0Hz, 1H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.84 (s, 1H), 4.36 (d, J =7.2Hz,1H),3.87-3.78(m,4H),3.75(d,J=3.4Hz,1H),3.73-3.61(m,2H ),3.58-3.40(m,6H),3.39-3.32(m,4H),3.28-3.15(m,2H),2.98(s,3H) ,2.87-2.76(m,1H),2.73-2.59(m,4H),2.54-2.38(m,4H),2.30(s,6H), 2.01-1.78(m,6H),1.71-1.64(m,1H),1.63-1.51(m,3H),1.48(s,3H),1 .46-1.37(m,3H),1.36-1.30(m,5H),1.29-1.25(m,4H),1.25-1.17(m,7 H), 1.01 (d, J = 7.4Hz, 3H), 0.94 (d, J = 7.0Hz, 3H), 0.86 (t, J = 7.4Hz, 3H). 13 C NMR(CDCl3,175MHz)δ:177.14,175.52,167.05,164.63,154.73,152.98,147.42,145.97,145.91,139.12,1 19.83,119.79,112.50,112.37,108.17,104.71,101.68,85.16,85.06,83.12,79.79,78.54,75.23,71.38,7 0.87,70.67,69.28,67.70,65.60,61.39,58.30,52.85,49.87,44.91,40.38,37.29,37.00,35.28,32.72,31.14,28.74,27.71,25.65,23.66,22.18,21.31,19.49,18.83,15.57,15.40,12.94,10.19,8.61,8.26,8.25.

[0182] Synthesis of target compound 34b

[0183]

[0184] According to the general synthetic method of this embodiment, intermediate compound 29 (0.147 g, 0.15 mmol) and enoxacin (Ar=b) (0.144 g, 0.45 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 34b (35 mg, 0.03 mmol, 20.0%). HRMS (ESI) (M+H) + m / z1077.6148, calcd for C 54 H 86 FN6O 15 1077.6130. 1 H NMR (CDCl3, 400MHz) δ: 8.69 (s, 1H), 8.09 (d, J = 13.4Hz, 1H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.84 (s, 1H), 4.47-4.31 (m, 3H), 3.94-3. 85(m,4H),3.85-3.77(m,4H),3.75(d,J=3.4Hz,1H),3.72-3.59(m,2H),3.56-3.36(m,5H),3.26-3.15(m,2H),2.98(s,3H),2.86-2.76(m,1 H),2.66-2.56(m,4H),2.51-2.38(m,4H),2.30(s,6H),2.03-1.79(m,6H),1.73-1.65(m,1H),1.65-1.54(m,3H),1.53-1.46(m,6H),1.46- 1.39(m,1H),1.36-1.30(m,4H),1.29-1.25(m,4H),1.25-1.19(m,6H),1.01(d,J=7.5Hz,3H),0.94(d,J=7.0Hz,3H),0.86(t,J=7.3Hz,3H). 13C NMR(CDCl3,175MHz)δ:177.09,177.08,175.51,167.00,164.60,154.74,150.46,150.40,148.10,146.36,14 5.10,120.28,120.15,113.76,113.74,109.32,101.66,85.16,85.05,83.13,79.82,78.54,75.26,71.37,70 .86,70.67,69.25,67.72,65.61,61.41,58.24,53.04,49.87,47.76,47.08,47.04,44.91,40.40,37.28,37.01,32.73,31.14,27.68,25.64,23.61,22.19,21.30,19.48,18.84,15.58,15.39,14.98,12.95,10.19,8.62.

[0185] Synthesis of target compound 34c

[0186]

[0187] According to the general synthetic method of this embodiment, intermediate compound 29 (0.153 g, 0.15 mmol) and norfloxacin (Ar=c) (0.151 g, 0.47 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5. The target compound 34c (36 mg, 0.03 mmol, 21.2%) was obtained. HRMS (ESI) (M+H) + m / z1076.6185, calcd for C 55 H 87 FN5O 15 1076.6177. 1H NMR (MeOD, 400MHz) δ: 8.76 (s, 1H), 7.88 (d, J = 12.3Hz, 1H), 7.05 (s, 1H), 5.07 (dd, J1 = 10.7Hz, J2 = 2.2Hz, 1H), 4.87 (s, 1H), 4.52-4.33 (m, 3H), 3.8 7-3.80(m,2H),3.80-3.76(s,3H),3.73-3.66(m,2H),3.58-3.44(m,5H), 3.42-3.32(m,4H),3.25-3.17(m,2H),2.97(s,3H),2.85-2.78(m,1H),2. 74-2.68(m,4H),2.65-2.57(m,2H),2.51-2.45(m,2H),2.41(s,6H),1.9 8-1.75(m,6H),1.65-1.60(m,4H),1.53-1.47(m,6H),1.45-1.38(m,2H), 1.32(s,3H),1.29-1.25(m,4H),1.23(d,J=6.1Hz,3H),1.17(d,J=6.8Hz, 3H), 1.00 (d, J = 7.5Hz, 3H), 0.95 (d, J = 6.9Hz, 3H), 0.86 (t, J = 7.3Hz, 3H). 13 C NMR(MeOD,175MHz)δ:175.68,165.52,101.35,85.37,85.19,83.11,78.88,7 8.58,74.97,71.39,70.90,70.30,68.57,67.17,64.41,60.21,58.02,52.66, 49.23,49.10,44.85,39.52,37.28,36.67,32.63,30.87,30.80,27.34,25.61,22.93,21.84,20.15,18.70,17.57,14.65,14.52,13.43,11.73,9.31,7.78.

[0188] Synthesis of target compound 35a

[0189]

[0190] According to the general synthetic method of this embodiment, intermediate compound 30 (0.193 g, 0.20 mmol) and ciprofloxacin (Ar=a) (0.199 g, 0.60 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 35a (38 mg, 0.04 mmol, 20.0%). HRMS (ESI) (M+H) + m / z1102.6346, calcd for C 57 H 89 FN5O 15 1102.6334. 1 H NMR (CDCl3, 400MHz) δ: 8.77 (s, 1H), 8.01 (d, J = 13.1Hz, 1H), 7.36 (d, J = 7.0Hz, 1H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.84 (s, 1H), 4.36 (d, J =7.2Hz,1H),3.87-3.78(m,4H),3.75(d,J=3.4Hz,1H),3.73-3.61(m,2H ),3.58-3.40(m,6H),3.39-3.32(m,4H),3.28-3.15(m,2H),2.98(s,3H) ,2.87-2.76(m,1H),2.73-2.59(m,4H),2.54-2.38(m,4H),2.30(s,6H), 2.01-1.78(m,6H),1.71-1.64(m,1H),1.63-1.51(m,5H),1.48(s,3H),1 .46-1.37(m,3H),1.36-1.30(m,5H),1.29-1.25(m,4H),1.25-1.17(m,7 H), 1.01 (d, J = 7.4Hz, 3H), 0.94 (d, J = 7.0Hz, 3H), 0.86 (t, J = 7.4Hz, 3H). 13C NMR(CDCl3,175MHz)δ:177.13,175.52,167.07,164.66,154.75,147.41,145.91,139.11,112.36, 108.16,104.72,101.65,85.15,85.08,83.12,79.79,78.53,75.21,71.39,70.98,70.67,69.26,6 7.69, 65.59, 61.39, 58.51, 52.89, 49.87, 44.91, 40.38, 37.28, 37.00, 35.28, 32.72, 31.13, 29.74, 28.72, 26.76, 25.65, 24.19, 22.18, 21.37, 19.49, 18.83, 15.57, 15.40, 12.95, 10.19, 8.61, 8.25.

[0191] Synthesis of target compound 35b

[0192]

[0193] According to the general synthetic method of this embodiment, intermediate compound 30 (0.149 g, 0.15 mmol) and enoxacin (Ar=b) (0.144 g, 0.45 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 35b (35 mg, 0.03 mmol, 20.0%). HRMS (ESI) (M+H) + m / z1091.6337, calcd for C 55 H 88 FN6O 15 1091.6286. 1H NMR (CDCl3, 400MHz) δ: 8.69 (s, 1H), 8.09 (d, J = 13.4Hz, 1H), 5.13 (dd, J1 = 10.8Hz, J2 = 2.5Hz, 1H), 4.84 (s, 1H), 4.47-4.31 (m, 3H), 3.94-3.85 (m ,4H),3.85-3.77(m,4H),3.75(d,J=3.4Hz,1H),3.72-3.59(m,2H),3.56 -3.36(m,5H),3.26-3.15(m,2H),2.98(s,3H),2.86-2.76(m,1H),2.66- 2.56(m,4H),2.53-2.44(m,2H),2.43-2.36(m,2H),2.30(s,6H),1.98- 1.80(m,6H),1.71-1.64(m,1H),1.62-1.54(m,5H),1.53-1.46(m,6H),1 .45-1.37(m,1H),1.36-1.30(m,4H),1.29-1.25(m,4H),1.25-1.19(m,6 H), 1.01 (d, J = 7.5Hz, 3H), 0.94 (d, J = 7.0Hz, 3H), 0.86 (t, J = 7.3Hz, 3H). 13 C NMR(CDCl3,175MHz)δ:177.07,175.51,167.02,164.64,154.75,150.44,150.39,148.09,146.35,145.09, 120.26,120.13,113.72,109.29,101.65,85.15,85.07,83.13,79.81,78.52,75.23,71.38,70.98,70.66, 69.24,67.70,65.58,61.40,58.44,53.08,49.87,47.77,47.07,47.02,44.91,40.38,37.28,37.00,32.72,31.13,29.74,26.71,25.65,24.14,22.19,21.30,19.49,18.83,15.57,15.39,14.98,12.95,10.19,8.62.

[0194] Synthesis of intermediate compound 36b

[0195]

[0196] According to the general synthetic method of this embodiment, intermediate compound 31 (0.221 g, 0.23 mmol) and enoxacin (Ar=b) (0.219 g, 0.68 mmol) were used as starting materials. Column chromatography conditions were 100-200 mesh silica gel. The mobile phase for the first column chromatography was dichloromethane / methanol / formic acid = 10 / 1 / 0.05, and the mobile phase for the second column chromatography was dichloromethane / methanol / ammonia = 10 / 1 / 0.5, yielding the target compound 36b (32 mg, 0.03 mmol, 13.1%). HRMS (ESI) (M+H) + m / z1077.6168, calcd for C 54 H 86 FN6O 15 1077.6130. 1 H NMR (MeOD, 400MHz) δ: 8.84 (s, 1H), 8.03 (d, J = 13.3Hz, 1H), 5.03 (s, 1H), 5.13 (m, 1H), 4.55-4.46 (m, 2H), 4.37-4.30 (m, 1H),4.26-4.21(m,1H),3.98-3.88(m,4H),3.79(s,3H),3.75-3.46(m,8H),3.41-3.36(m,1H),3.23-3.16(m,1H),3.03- 2.94(m,1H),2.74(s,6H),2.71-2.63(m,6H),2.51-2.44(m,2H),2.16-2.07(m,1H),2.02-1.82(m,4H),1.66-1.60(m,4H ),1.57(s,3H),1.51-1.45(m,5H),1.32-1.26(m,10H),1.19(d,J=6.6Hz,3H),1.12(d,J=7.0Hz,3H),0.98-0.91(m,6H). 13 C NMR(CDCl3,175MHz)δ:176.89,172.77,168.68,165.60,154.95,147.28,145.23 ,119.26,119.13,113.29,101.52,85.77,83.07,78.06,75.68,70.28,69.40,68. 12,67.12,65.19,60.19,57.88,52.62,46.45,38.70,37.75,34.84,32.71,30.44,30.00,27.28,25.88,22.80,19.93,18.58,17.56,14.36,13.85,12.13,10.17.

[0197] The target compound of this application—a macrolide derivative—can be prepared according to the above preparation examples. The macrolide is connected to a quinolone via an ether side chain at the 3-position. Compared with derivatization via an ester group at the 3-position, this greatly improves the stability of the prepared macrolide derivative.

[0198] Furthermore, to demonstrate the antibacterial properties of the target compounds prepared in this application, and the relationship between the dual-target compounds prepared above and the antibacterial properties, the following examples are provided.

[0199] Example 1: In vitro antibacterial activity test of the target compound

[0200] According to the CLSI guidelines (Clinical and Laboratory Standards Institute, CLSI), the in vitro antibacterial activity of some target compounds against susceptible Streptococcus pneumoniae ATCC49619, Streptococcus pneumoniae PU09 (mef-resistant), Streptococcus pneumoniae 07P390 (constitutive ermB-resistant), Streptococcus pyogenes 12-206 (constitutive ermTR-resistant), and Streptococcus pyogenes 01-968 (inducible ermB-resistant) was determined using the broth dilution method. Each bacterial strain was plate-transferred and purified before the test, and fresh cells were used for the experiment. A standard strain was used as the test control for each experiment; antibiotic-free bacterial suspensions were used as growth controls for the test strains. The minimum inhibitory concentration (MIC) was determined using the broth two-fold dilution method. The concentration range for antibiotic determination was 256–0.008 μg / mL, and the final concentration of the test bacterial suspension was 5 × 10⁻⁶. 5 CFU / mL. The reference drugs were commercially available macrolides (solimin, clarithromycin, ciprofloxacin, erythromycin, telithromycin).

[0201] The test results are shown in Tables 1 and 2:

[0202] Table 1: Antibacterial activity (MIC, μg / mL) of target compounds 20-24, 26, 32a, and 33a

[0203]

[0204]

[0205] Table 2: Antibacterial activity of target compounds 25 and 32-36 (MIC, μg / mL)

[0206]

[0207] As can be seen from the test results in Tables 1 and 2, the target compound prepared in this application exhibits better antibacterial activity compared to the control drug. Specifically, the antibacterial activity of target compound 33b is significantly improved compared to all commercially available structurally similar macrolide compounds: erythromycin, clarithromycin, and telithromycin, particularly against susceptible Streptococcus pneumoniae ATCC49619, Streptococcus pneumoniae PU09 (mef-resistant), Streptococcus pyogenes 12-206 (constitutive ermTR-resistant), and Streptococcus pyogenes O1-968 (inducible ermB-resistant). For Streptococcus pneumoniae O7P390 (constitutive ermB-resistant), target compound 33b shows better antibacterial activity than erythromycin and clarithromycin, and similar antibacterial activity to telithromycin.

[0208] Table 2 compares the antibacterial activity data of the published similar compound 19G. The target compound 33b of this application shows a significant improvement in antibacterial activity against susceptible Streptococcus pneumoniae ATCC49619, Streptococcus pneumoniae PU09 (mef-resistant), and Streptococcus pyogenes 01-968 (inducible ermB resistance), and also restores antibacterial activity against constitutively ermB-resistant Streptococcus pneumoniae 07P390 (MIC: 33b = 1 μg / mL; 19G > 32 μg / mL). This indicates that the target compound 33b has better structural and antibacterial activity advantages.

[0209] This application describes a macrolide 3-OH ether hydrocarbon derivative obtained by linking a quinolone group to the 3-position of a macrolide with an ether hydrocarbon. The ether hydrocarbon side chain improves the stability of the macrolide derivative, and the dual-target property enhances its antibacterial activity and restores its activity against constitutively resistant bacteria.

[0210]

[0211] (Compound 19G in Table 2 corresponds to compound 19G in Eur J Med Chem, 2024:116630)

[0212] Example 2: DNA helicase inhibition experiment

[0213] The target compound's half-maximal inhibitory concentration (IC50) of E. coli gyrase 50The results were determined using the Inspiralis (Norwich, UK) gyrase supercoiling kit. The supercoiling reaction mixture consisted of 35 mM Tris.HCl (pH 7.5), 24 mM KCl, 4 mM MgCl2, 2 mM DTT, 1.8 mM Spermidine, 1 mM ATP, 6.5% (w / v) glycerol, 0.1 mg / mL BSA, 1 U DNA gyrase, 0.5 μg relaxed pBR322 DNA, and target compound 33b at concentrations of 50, 25, 12.5, 6.25, and 3.125 μM, and incubated at 37 °C for 30 min. Ciprofloxacin (CIP) was used as a gyrase inhibitor control. Each reaction was stopped by adding 30 μl of stop buffer and 30 μl of chloroform / isoamyl alcohol (24 / 1). The DNA products were analyzed by electrophoresis on a 1% agarose gel in TAE buffer. After staining with 0.5 μg / mL ethidium bromide, the gel was photographed and quantified using a ChemiScope 6100 (Clinx, CN).

[0214] See Figure 1 , Figure 1 This is a diagram showing the results of a DNA gyrase supercoiling experiment of target compound 33b provided in an embodiment of this application. Figure 1 Part (a) is an electrophoresis diagram of the DNA gyrase supercoiling experiment. Figure 1 In section (b), from left to right, are the IC50 values ​​of target compound 33b and ciprofloxacin, respectively. 50 Value. From Figure 1 As shown in section (a), the substrate pBR322 is in a relaxed state in the absence of DNA gyrase, while it becomes supercoiled after the addition of DNA gyrase; in the system with 50 μM of target compound 33b, the substrate is essentially in a relaxed state. Figure 1 As can be seen in section (b), the IC50 of target compound 33b is... 50 The value was 24.190±5.911 μM, the IC50 value of ciprofloxacin. 50 The value was 0.218 ± 0.014 μM, indicating that the target compound 33b has the ability to inhibit topoisomerase.

[0215] Example 3: Protein synthesis inhibitory activity IC 50 Measurement experiment

[0216] The materials used in the *E. coli* in vitro transcription / translation (TnT) assay were derived from the *E. coli* S30 loop DNA extraction system (catalog number L1020, Promega, USA). This kit contains all the components required for efficient transcription and translation of user-supplied DNA templates. To measure protein synthesis inhibition, the assay used the plasmid pBESTluc containing the firefly luciferase gene as the DNA template. The luminescence intensity produced by luciferase expression corresponds to the efficiency of protein synthesis. The luminescence intensity was quantified in the presence or absence of the test compound. Target compound 33b was evaluated at concentrations of 5, 1, 0.5, 0.1, and 0.01 μM to determine its 50% inhibitory concentration (IC50). 50 The value was compared with commercially available erythromycin (ERY) as a control.

[0217] See Figure 2 , Figure 2 This application provides an embodiment of the target compound 33b with inhibitory activity against protein synthesis (IC). 50 The assay results show that commercially available erythromycin is a single-target drug that inhibits protein synthesis, and its IC50 inhibitory activity against protein synthesis is shown in the figure. 50 The value was 0.40 ± 0.006 μM, and the IC50 value of the target compound 33b for its inhibitory activity against protein synthesis was 0.40 ± 0.006 μM. 50 The value was 0.65±0.031 μM, which is similar to the inhibitory effect of commercially available erythromycin on protein synthesis, indicating that the target compound 33b has a significant inhibitory effect on protein synthesis.

[0218] The experimental results of Examples 2 and 3 show that the target compound 33b prepared in this application simultaneously inhibits the synthesis of topoisomerases and proteins, indicating that the target compound 33b prepared in this application is a compound with dual targets, acting on both ribosomes and topoisomerases. Its antibacterial activity results also show this (Tables 1 and 2). The antibacterial activity of the dual-target target compound 33b is significantly superior to that of erythromycin, telithromycin, clarithromycin, and ciprofloxacin, which only have single-target inhibitory effects (the targets of erythromycin, telithromycin, and clarithromycin are ribosomes, while the target of ciprofloxacin is topoisomerase). Furthermore, the target compound 33b of this application restored the antibacterial activity against constitutively ermB-resistant Streptococcus pneumoniae 07P390 (MIC: 33b = 1 μg / mL; 19G > 32 μg / mL).

[0219] Example 4: Experiment with engineered Escherichia coli

[0220] According to CLSI guidelines, the MIC of the target compound against the SQ110DTC series of engineered bacteria was determined using the microbroth dilution method. The compound and positive control were diluted 1 / 2 times with broth in sterile 96-well plates, and then mixed with 100 μL of CAMHB broth containing the bacteria (approximately 1 × 10⁻⁶). 6 Mix (CFU / mL). Maintain the final concentration of DMSO below 1% during the measurement process. Finally, incubate the 96-well plate at 37°C for 16–20 hours, observe and record the results. MIC measurements should be repeated at least three times.

[0221] The determination results of target compound 35a are shown in Table 3:

[0222] Table 3: Inhibitory activity of target compound 35a against engineered Escherichia coli (MIC, μg / mL)

[0223]

[0224] a SQ110DTC: Escherichia coli with tolc deficiency; b SQ110DTC A2058G: A2058G mutation in 23 rRNA; c SQ110DTC gyrA306: C248T mutation in gyrA; d SQ110DTC A2058G / gyrA306: Double mutation (A2058G mutation in 23rRNA and C248T mutation in gyrA).

[0225] As shown in Table 3, compared with SQ110DTC, the MIC of target compound 35a against SQ110DTC A2058G remained unchanged; when gyrA306 was mutated, the MIC increased by 1 time; while when both A2058G and gyrA306 were mutated, the MIC ratio was greater than 16, and the antibacterial activity decreased significantly, indicating that target compound 35a can act on both ribosomes and topoisomerases. The antibacterial activity (Tables 1 and 2) shows that the antibacterial activity of target compound 35a is far superior to that of erythromycin, telithromycin, clarithromycin, and ciprofloxacin (the targets of erythromycin, telithromycin, and clarithromycin are ribosomes, while the target of ciprofloxacin is topoisomerases), which only have single-target inhibitory activity.

[0226] The above provides a detailed description of a macrocyclic lactone derivative, its preparation method, and its application. Specific examples have been used to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this application. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this application. Therefore, the content of this specification should not be construed as a limitation of this application.

Claims

1. A macrocyclic lactone derivative, characterized in that, The macrocyclic lactone derivative is a compound having the general formula shown in Formula I: Formula I In the formula, A is selected from any one of alkynyl, alkenyl or piperazine; R is selected from any one of H or F; X is selected from any one of CH or N; Y is selected from any one of methyl, ethyl or cyclopropyl; and n is an integer ranging from 2 to 6.

2. The macrocyclic lactone derivative according to claim 1, characterized in that, A is selected from piperazine or alkynyl, R is selected as F, X is selected from CH or N, Y is selected from ethyl or cyclopropyl, and n is an integer ranging from 3 to 5.

3. The macrocyclic lactone derivative according to claim 1, characterized in that, A is selected as piperazine, R as F, X as N, Y as ethyl, and n is 3. A is selected as piperazine, R as F, X as CH, Y as cyclopropyl, and n is 5.

4. The structure of the macrocyclic lactone derivative as described in claim 1 is shown below: 。 5. A method for preparing a macrocyclic lactone derivative, characterized in that, The method includes: introducing different side chains into the compound shown in formula (1), introducing quinolone groups through a reaction, and removing the protecting groups to obtain the compound shown in formula I; Equation (1) Equation I In the formula, A, R, X, Y, and n independently have the definitions of any one of claims 1-4.

6. The preparation method according to claim 5, characterized in that, The method includes a first synthesis, which is used to synthesize a compound of formula II in which A is selected as ethynyl, R as H, X as CH, and Y as methyl. Formula (1) Formula (2) Formula II In the first synthesis method, the compound shown in formula (1) is introduced with alkynol side chains of different lengths to obtain the compound shown in formula (2). The compound shown in formula (2) is introduced with quinolone groups through a sonogashira coupling reaction. After removing the protecting group, the compound shown in formula II is obtained. In the compound shown in formula II, the value of n is an integer ranging from 2 to 6.

7. The preparation method according to claim 5, characterized in that, The method includes a second synthesis method, which is used to synthesize the compound shown in Formula III where A is selected as vinyl, R as H, X as CH, Y as methyl, and n is 4. Formula (1) Formula (3) Formula III In the second synthesis, the compound shown in formula (1) is introduced with an enol side chain to obtain the compound shown in formula (3). The compound shown in formula (3) is introduced with a quinolone group through a Heck coupling reaction, and after removing the protecting group, the compound shown in formula III is obtained.

8. The preparation method according to claim 5, characterized in that, The method includes a third synthesis, which is used to synthesize the compound of formula IV, where A is selected as piperazine and R is selected as F: Formula (1) Formula (4) Formula IV In the third synthesis, the compound shown in formula (1) is introduced with diol side chains of different lengths to obtain the compound shown in formula (4). The compound shown in formula (4) is sulfonated and then a quinolone group is introduced through a nucleophilic substitution reaction. The protecting group is removed to obtain the compound shown in formula IV. In the compound shown in formula IV, X, Y, and n independently have the definitions of any one of claims 1-4.

9. The use of at least one of the macrolide derivatives or pharmaceutically acceptable salts thereof as described in any one of claims 1-4 in the preparation of a medicament, wherein the medicament is an antimicrobial drug and the pathogenic microorganism is bacteria.

10. The use of at least one of the macrolide derivatives or pharmaceutically acceptable salts thereof as described in any one of claims 1-4 in the preparation of an antibiotic; said antibiotic being used to inhibit the activity of Streptococcus pneumoniae and / or Streptococcus pyogenes.

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