CN111072740A

CN111072740A - Erythromycin A ketolide antibiotic derivative, and preparation method and application thereof

Info

Publication number: CN111072740A
Application number: CN201811216765.3A
Authority: CN
Inventors: 赵哲辉; 雷平生; 张晓曦; 杨爽
Original assignee: Institute of Materia Medica of CAMS
Current assignee: Institute of Materia Medica of CAMS
Priority date: 2018-10-18
Filing date: 2018-10-18
Publication date: 2020-04-28
Anticipated expiration: 2038-10-18
Also published as: CN111072740B

Abstract

The invention discloses a series of erythromycin A ketolide antibiotics containing aminoquinoline rings as shown in formula I, a preparation method and application thereof, and side chain intermediates and a synthesis method of each compound. The key points of the invention are as follows: the compound shown in the formula I has the drug effect of broad-spectrum antibiotics, has outstanding antibacterial activity on sensitive and drug-resistant gram-positive bacteria and gram-negative bacteria, and particularly has antibacterial activity obviously superior to that of a control drug of telithromycin on vancomycin-resistant enterococcus faecalis, enterococcus faecium, erythromycin-resistant streptococcus pneumoniae, streptococcus pyogenes, klebsiella pneumoniae, shigella flexneri and other strains. The compound provided by the invention can be used as a broad-spectrum antibiotic and has the advantages ofInhibiting antibacterial and antiviral activity of gram-positive bacteria and gram-negative bacteria.

Description

Erythromycin A ketolide antibiotic derivative, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines. The invention relates to a series of erythromycin A ketolide antibiotic derivatives containing amino-quinoline substitution and having high antibacterial activity, a synthetic method thereof, a related intermediate synthetic method, biological activity of the compounds, and application of the compounds as broad-spectrum antibiotics in inhibiting sensitive and drug-resistant gram-positive bacteria and gram-negative bacteria and resisting viruses.

Background

Erythromycin A is the first macrolide antibiotic in clinical application, has definite curative effect, lower toxicity and good safety, is widely used for treating infection caused by gram-positive bacteria including staphylococcus aureus, staphylococcus epidermidis, streptococcus pneumoniae, streptococcus pyogenes and the like, and also has good antibacterial effect on atypical pathogenic bacteria such as chlamydia, mycoplasma, legionella, helicobacter pylori and the like, but with the long-term and wide use of antibiotics, drug-resistant bacteria of macrolide antibiotics are continuously discovered.

Ketolide is a third-generation macrolide antibiotic, and overcomes the problem of induced resistance of the first two generations of macrolide antibiotics while maintaining good antibacterial activity against non-resistant bacteria. Telithromycin is the first approved ketolide antibiotic, has broad-spectrum antibacterial activity, and has outstanding antibacterial activity on penicillin and erythromycin resistant streptococcus pneumoniae. However, with the clinical application of telithromycin, the hepatotoxicity is reported increasingly, and a plurality of studies show that although the incidence rate of serious adverse hepatic events of telithromycin is low, acute hepatic failure and death due to serious hepatic injury are reported. For safety reasons, the FDA in the united states limits the indications of telithromycin, affecting the clinical use of telithromycin. It is therefore essential to accelerate the development of novel antibiotics with broad-spectrum antibacterial activity, in particular to enhance antibacterial activity against drug-resistant bacteria and to reduce the side effects such as hepatotoxicity of the drugs.

Many researchers have performed much work on modification of the macrocyclic ketolide structure. Nomura T et al oximate the 9-position of ketolide, introduced acetamido-aromatic ring side chain, at the same time, 11, 12-position form cyclic carbonate, the results of in vitro activity test found that when the introduced aromatic ring is quinoline ring, the antibacterial activity is the best, the antibacterial activity to Streptococcus pneumoniae and Haemophilus influenzae is obviously better than that of the control drug telithromycin.

Ly T. Phan et alThe 4' -mycaminose in the 5-site dimethyl aminose of the hexadecanolide is cut off, then a quinoline ring side chain is introduced, and the activity data shows that the antibacterial activity of the compound to sensitive bacteria and drug-resistant bacteria is improved. The authors suggest that the quinoline ring side chain may be closer to the peptidyl transferase center and thus better inhibit bacterial protein synthesis. Simultaneously, quinoline rings are introduced to generate pi-pi accumulation or hydrophobic interaction with ribosome bases, so that the affinity of drug molecules and bacterial ribosome is increased, and the activity of resisting drug-resistant bacteria is improved^[39]。

The compounds of the application are ketolide derivatives which contain fluorine at the 2-position and have aminoquinoline side chain substitution at the 11, 12-positions, and in-vitro antibacterial activity tests are carried out on the compounds. Has the advantages of simple and efficient synthetic route, and can obtain the target product with high yield by taking the commercialized amino-quinoline as the starting material through simple reaction. Meanwhile, the obtained derivative has outstanding antibacterial activity, the antibacterial activity of most compounds is equivalent to that of telithromycin, the antibacterial activity of part of compounds to sensitive bacteria and drug-resistant bacteria is superior to that of telithromycin, and the problem of bacterial drug resistance is hopeful to be solved.

Disclosure of Invention

The invention aims to provide macrocyclic ketolide compounds with broad-spectrum antibacterial activity, namely erythromycin A macrocyclic ketolide antibiotic derivatives with a new structure and pharmaceutically acceptable salts thereof, and provides a preparation method, a pharmaceutical composition and application thereof in preparation of antibacterial or viral medicaments.

In order to solve the technical problem, the invention provides the following technical scheme:

the first aspect of the technical scheme of the invention provides macrocyclic ketolide antibiotic compounds shown as a formula I and pharmaceutically acceptable salts thereof, wherein the macrocyclic ketolide antibiotic compounds have the following structural formula:

x is selected from hydrogen, C1-6 alkyl, C6-10 aryl C1-4 alkyl, C1-6 alkanoyl, C6-10 aryl C1-4 alkanoyl and cinnamoyl;

y is selected from substituted or unsubstituted quinoline ring, wherein the substituent is selected from hydrogen, amino, nitro, C1-3 alkoxy.

Preferred compounds are shown as IA and are characterized in that,

wherein X is selected from hydrogen, C1-6 alkyl, C6-10 aryl C1-4 alkyl, C1-6 alkanoyl, C6-10 aryl C1-4 alkanoyl and cinnamoyl;

preferred compounds are represented by formula IB, wherein,

the above-mentioned C1-6 alkyl group and C1-6 alkane in the C1-6 alkanoyl group specifically include: c1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl;

preferred compounds are specifically listed below: compound S1

Compound S2

Compound S3

Compound S4

Compound S5

Compound S6

Compound S7

Compound S8

Compound S9

Compound S10

Compound S11

Compound S12

Compound S13

The second aspect of the technical scheme of the invention is to provide a preparation method of the macrocyclic ketolide antibiotic derivative with the novel structure in the first aspect. Wherein the starting material is clarithromycin, and a proper intermediate is obtained by adopting a reaction method reported by a literature or feasible in synthetic chemistry and is used as the raw material of the invention, and the structural formula is as follows:

the synthesis method of the raw material 8 in the invention is as follows:

reaction reagents and conditions: a. hydrochloric acid, water, room temperature, 4 hours, 83.8%; b. acetic anhydride, dichloromethane, room temperature, 92.7%; c. triphosgene, pyridine, dichloromethane, 0 ℃, 85.3%; d. dess-martin oxidant, dichloromethane, 0 ℃, 97.5%; e.1-bisoxazolbicyclo [5,4,0] -7-undecene, acetone, 70 ℃ 82.6%; f, N, N-carbonyldiimidazole, sodium hydride, N, N-dimethylformamide, 90.5% at-20 ℃, h, methanol, 12h, 83.1%.

The compound synthesis method comprises the steps of starting from clarithromycin, and obtaining a macrolide intermediate compound 8 which is properly protected through a series of reactions; then, starting with the corresponding amino-quinoline, firstly modifying the amino, and then extending the carbon chain to obtain a primary amine side chain substituted by the selected amino-quinoline ring; then, the macrolide intermediate compound 8 and a primary amine side chain are subjected to addition reaction to obtain a corresponding erythromycin A antibiotic derivative;

wherein X, Y is defined as the first aspect of the present invention.

According to a third aspect of the present invention, there is provided a pharmaceutical composition comprising at least one compound of the amino-quinoline substituted erythromycin A ketolide antibiotic derivatives of the first aspect and a pharmaceutically acceptable carrier. The pharmaceutical composition may be prepared according to methods well known in the art. The compounds of the invention may be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The compounds of the present invention are generally present in the pharmaceutical compositions in an amount of from 0.1 to 95% by weight.

The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form by enteral or parenteral routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oromucosal, ophthalmic, pulmonary and respiratory, dermal, vaginal, rectal, and the like.

The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The compound can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle drug delivery systems.

For tableting the compounds of the invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like.

The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.

To encapsulate the administration units, the active ingredient of the compounds of the invention can be mixed with diluents and glidants and the mixture can be placed directly into hard or soft capsules. Or the effective component of the compound of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants, glidants used to prepare the compound tablets of the present invention may also be used to prepare capsules of the compound of the present invention.

In order to prepare the compound of the invention into injection, water, ethanol, isopropanol, propylene glycol or the mixture thereof can be used as a solvent, and a proper amount of solubilizer, cosolvent, pH regulator and osmotic pressure regulator which are commonly used in the field are added, wherein the solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl- β -cyclodextrin and the like, the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide and the like, the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate and the like, and the mannitol, glucose and the like can be added as a propping agent when preparing freeze-dried powder injection.

In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.

For the purpose of administration and enhancing the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is from 0.001 to 150mg/Kg body weight, preferably from 0.1 to 100mg/Kg body weight, more preferably from 1 to 60mg/Kg body weight, and most preferably from 2 to 30mg/Kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.

The compounds or compositions of the present invention may be administered alone or in combination with other therapeutic or symptomatic agents. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.

According to a fourth aspect of the present invention there is provided the use of a compound according to the first aspect of the present invention in the manufacture of a medicament for inhibiting bacteria.

The gram-positive bacteria are selected from staphylococcus aureus, streptococcus pyogenes, staphylococcus epidermidis, streptococcus pneumoniae, enterococcus faecalis and enterococcus faecium;

the gram-negative bacteria are selected from Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter cloacae, Serratia marcescens, Citrobacter fraudulans, Prevotella farinosa, Proteus mirabilis, Pseudomonas maltophilia and Shigella flexneri.

Advantageous technical effects

The antibiotic derivatives of the macrolide compound erythromycin A containing the side chain substituted by the amino-quinoline ring derivatives in the invention have the in vitro antibacterial activity reaching the level equivalent to that of telithromycin, and are superior to telithromycin for part of strains, and the sample compound designed and synthesized by the invention avoids the side chain structure which can cause hepatotoxicity; compared with telithromycin synthesis, the sample compound related by the invention has the advantages of short synthetic route, simple operation, high total yield and low cost. Therefore, the drug candidate with novel structure, strong activity and simple synthesis is provided, and the drug candidate can be used as a broad-spectrum antibiotic to inhibit gram-positive bacteria and gram-negative bacteria and resist viruses.

Detailed Description

The present invention will be further described by the following examples, however, the scope of the present invention is not limited to the following examples. It will be understood by those skilled in the art that various changes and modifications may be made to the invention without departing from the spirit and scope of the invention. The present invention has been described generally and/or specifically with respect to materials used in testing and testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible. For all of the following examples, standard procedures and purification methods known to those skilled in the art may be used. Unless otherwise indicated, all temperatures are expressed in degrees Celsius. The structure of the compounds is determined by nuclear magnetic resonance spectroscopy (NMR) and/or Mass Spectrometry (MS).

Preparation example section

The structure of the compound is shown by nuclear magnetic resonance hydrogen spectrum (¹H NMR), nuclear magnetic resonance carbon spectrum (C¹³C NMR) and Mass Spectrometry (MS). The hydrogen and carbon spectral shifts (δ) for nuclear magnetic resonance are given in parts per million (ppm). NMR spectra were obtained using a NMR spectrometer model Mercury-300, Mercury-400, Bruke-400 or Mercury-600, deuterated chloroform (CDCl)₃) Or heavy water (D)₂O) or deuterated dimethyl sulfoxide (DMSO-d)₆) As solvent Tetramethylsilane (TMS) was used as internal standard.

The high resolution mass spectrum is measured by an Agilent 1100series LC/MSD trap mass spectrometer or a Theromo active orbitrap plus LC/MSD mass spectrometer.

The column chromatography generally uses 160-200 mesh silica gel as a carrier.

The anhydrous solvents were all processed by standard methods. Other reagents were all commercially available analytical grade.

Wherein the content of the first and second substances,

ac acetyl

Bn benzyl

brs broad single broad singlet

Bz benzoyl

CDI 1,1 '-Carbonyldiimidazole 1, 1' -carbonyldiimidazole

double d double

DBU 1,8-diazabicyclo [5,4,0] -7-ene 1, 8-dioxazabicyclo [5,4,0] -7-undecene

DMF dimethyl formamide N, N-dimethylformamide

DMSO dimethyl sulfoxide

equivalent of eq, equivalent

Et ethyl

ESI electrospray ionization

gram

h hour of hour

Hz hertz

m multiple of

m/z mass to charge ratio

Me methyl group

Microliter of microliter

mg milligram

Minimum inhibitory concentration for MIC minimum inhibitory concentration

ml millilite ml

mmol millimole

MS mass spectrometry

NMR nuclear magnetic resonance

ppm part per millionth

pyridine

quadruple q quater

ref reference literature

rt room temperature

triple of t-triples

THF tetrahydrofuran

TLC thin layer chromatography

Preparation example 1 preparation of starting Compound 8

The macrocyclic ketolide compound with the novel structure is prepared from clarithromycin by adopting a feasible reaction method which is reported in a known literature or known by the public in synthetic chemistry, and an appropriate derivative is obtained to be used as a raw material of the invention.

2.0g, 2.675mmol of clarithromycin was added to 20ml of 1N hydrochloric acid under cooling in an ice-water bath and stirred, and when the reaction started, the reaction solution was viscous and stirred for 4 hours, and the reaction solution gradually became clear. TLC (CH)₂Cl₂:CH₃OH ═ 10:1) the reaction was monitored for completion, i.e. stopped. And (2) cooling the reaction system in an ice-water bath, adding a 20% sodium hydroxide solution into the reaction solution while stirring, adjusting the pH to be alkaline, separating out a large amount of white solid, filtering, washing the filter cake for multiple times, and drying to obtain a white powdery solid 1(1.371g, 83.5%).

¹H NMR(300MHz,CDCl₃):δ5.17(dd,1H,J＝2.1Hz,J＝10.8Hz),4.38(d,1H,J＝7.2Hz),3.91(s,1H),3.85(s,1H),3.68(s,1H),2.96(s,3H),2.26(s,6H),2.11(q,1H,J＝7.5Hz),1.36(s,3H),1.17(s,3H),0.83(t,3H,J＝7.2Hz).

¹³C NMR(75MHz,CDCl₃):221.11,175.41,107.02,88.74,79.39,78.45,74.60,71.06,70.68,70.18,66.08,49.97,45.93,44.97,40.66,39.15,37.93,36.28,28.45,21.84，21.67,19.17,18.15,16.60,15.60,13.05,10.83,8.64.

HR-MS(ESI)(M+H)⁺m/z 590.3885, calculated: c₃₀H₅₆NO₁₀590.3898.

Dissolving 2.05g of 3.478mmol of compound 1 in 20ml of anhydrous dichloromethane, cooling the reaction solution in an ice-water bath environment, adding 0.4223g of 4.1736mmol and 0.58ml of triethylamine into the reaction solution after the temperature is stabilized, fully stirring, slowly dropwise adding 0.4261g of acetic anhydride, 4.136mmol and 0.39ml of acetic anhydride into the reaction solution, and stirring for overnight reaction. TLC (CH)₂Cl₂:CH₃OH ═ 10:1) the reaction was monitored for completion, i.e. stopped. The reaction mixture was diluted to 50ml with dichloromethane, washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried to give 2(2.086g, 93.6%) as a white foamy solid.

¹H NMR(300MHz,CDCl₃):δ5.12(d,1H,J＝9.0Hz),4.74～4.71(m,2H),4.57(d,1H,J＝7.5Hz),3.69(s,1H),3.51～3.45(m,2H),2.91(s,3H),2.74～2.64(m,2H),2.60～2.54(m,1H),2.25(s,6H),2.05(s,3H),1.48(s,3H),1.18(d,3H,J＝6.6Hz),1.13(d,3H,J＝7.5Hz),0.93(d,3H,J＝7.2Hz),0.86(t,3H,J＝7.5Hz).

¹³C NMR(75MHz,CDCl₃):212.52,175.28,170.27,154.45,100.30,85.25,81.59,81.28,78.52,78.15,75.65,71.92,69.31,63.59,50.00,45.65,44.54,41.07,38.96,37.82,36.30,31.35,22.51,21.86,21.52,19.67,18.70,15.63,14.51,13.43,10.52,8.15.

HR-MS(ESI)(M+H)⁺m/z 658.3815, calculated: c₃₃H₅₆NO₁₂658.3797.

2.53g of compound 3 (4.007 mmol) is dissolved in 25ml of anhydrous dichloromethane, 6ml of pyridine is added into the reaction solution, the reaction system is placed in an ice water bath for cooling, after the temperature is stabilized, 2.973g of triphosgene dichloromethane solution (15ml) is added dropwise through a constant pressure dropping funnel, and the color of the reaction solution is deepened until the reaction solution becomes dark brown during the addition of the triphosgene. The reaction system was gradually returned to room temperature, and after stirring for 4 hours, the completion of the reaction was monitored by TLC (petroleum ether: acetone ═ 3:1) and the reaction was stopped. Diluting the reaction solution to 100ml with dichloromethane, injecting into a separating funnel, slowly dropping the reaction solution into 100ml saturated sodium bicarbonate solution, paying attention to heat release and air release, simultaneously paying attention to supplement sodium bicarbonate solid, adjusting pH to alkalinity, standing for layering, separating an aqueous phase and an organic phase, washing the organic phase with 100ml of 5% potassium dihydrogen phosphate solution and saturated salt water for 1 time respectively, paying attention to emulsification, separating the organic phase, discharging an emulsion layer, and discarding the aqueous phase. Diluting the emulsion layer with 1 volume of water and dichloromethane, and standing for layering. And combining organic phases, drying by anhydrous sodium sulfate, filtering, spin-drying, separating by column chromatography, infiltrating by petroleum ether, and mixing the organic phases: ethyl acetate 1: 1-pure ethyl acetate, the fractions were combined and spun dry to give 3 as a white foam solid (2.599g, 83.7%).

¹H NMR(300MHz,CDCl₃):δ5.12(d,1H,J＝9.0Hz),4.77～4.71(m,2H),4.57(d,1H,J＝7.5Hz),3.69(s,1H),3.50～3.45(m,2H),2.91(s,3H),2.74～2.64(m,2H),2.60～2.54(m,1H),2.25(s,6H),2.05(s,3H),1.48(s,3H),1.18(d,3H,J＝6.6Hz),1.13(d,3H,J＝7.5Hz),0.93(d,3H,J＝7.2Hz),0.86(t,3H,J＝7.5Hz).

HR-MS(ESI)(M+H)⁺m/z 658.3815, calculated: c₃₃H₅₆NO₁₂658.3797.

4.652g, 7.077mmol of Compound 3 were dissolved in 45ml of anhydrous dichloromethane, the reaction system was cooled in an ice-water bath, 3.062g, 8.492mmol of dess-martin oxidant were added in portions and stirred to dissolve it sufficiently. The reaction solution was gradually returned to room temperature, and after further reaction for 4 hours, TLC (CH)₂Cl₂:CH₃OH ═ 10:1) the reaction was monitored for completion, i.e. stopped. Adding 45ml of 10% sodium thiosulfate solution into the reaction solution, stirring to fully mix the sodium thiosulfate solution with the reaction solution, standing for 15min for layering, wherein a large amount of white flocculent precipitate appears, filtering, discarding white solid, separating liquid, discarding an aqueous phase, washing an organic phase with 45ml of saturated sodium bicarbonate for 1 time, washing with saturated salt for 1 time, drying the organic phase with anhydrous sodium sulfate, filtering, and evaporating to dryness. Separating by column chromatography, soaking with petroleum ether, eluting with ethyl acetate, mixing the components, and evaporating to dryness to obtain white foamy solid 4(4.499g, 97.6%).

¹HNMR(300MHz,CDCl₃):δ5.05(dd,1H,J＝2.7Hz,J＝9.9Hz),4.73(dd,1H,J＝10.2Hz,J＝8.1Hz),4.62(s,1H),4.37(d,1H,J＝7.8Hz),4.16(d,1H,J＝7.5Hz),3.78(q,1H,J＝6.9Hz),3.54(m,1H),3.02～2.94(m,2H),2.64(s,3H),2.24(s,6H),2.04(s,3H),1.94(s,1H),1.86～1.81(m,1H),1.73～1.70(m,1H),1.54(s,3H),1.47(s,3H),1.42～1.40(m,3H),1.36～1.34(m,3H),1.31(s,3H),1.26～1.22(m,3H),1.15～1.12(m,6H),0.89(t,3H,J＝7.5Hz).

¹³C NMR(75MHz,CDCl₃):213.27,204.41,170.19,169.49,154.29,101.87,84.89,81.31,78.76,78.57,77.12,72.00,69.60,63.84,51.52,49.93,47.97,44.22,41.08,39.67,38.52,30.85,22.78,21.83,21.42,20.10,18.31,16.61,14.52,13.97,12.90,10.77.

HR-MS(ESI)(M+H)⁺m/z 656.3627,calcd for C₃₃H₅₄NO₁₂656.3641.

5.27g, 8.041mmol of Compound 4 were dissolved in 75ml of acetone, 1.8ml, 1.835g, 12.06mmol of 1, 8-diazabicycloundec-7-ene (DBU) were added to the reaction system, heated to 65 ℃ and reacted for 4h, followed by TLC (CH)₂Cl₂:CH₃OH ═ 10:1) the reaction was monitored for completion, i.e. stopped. The reaction system was cooled to room temperature, acetone was removed under reduced pressure, the residue was dissolved in 150ml of dichloromethane, washed 1 time with 75ml of 5% potassium dihydrogen phosphate solution and saturated brine, respectively, the organic phase was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was slurried with 25ml of ethyl acetate overnight and filtered to give a light brown powdery solid 5(4.17g, 85.6%).

¹H NMR(300MHz,CDCl₃):δ6.59(s,1H),4.98(d,1H,J＝9.6Hz),4.71(t,J＝8.0Hz1H),4.34(d,1H,J＝8.0Hz),4.13(d,1H,J＝8.0Hz),3.72(q,1H,J＝6.8Hz),3.54～3.50(m,1H),3.16(q,1H,J＝6.0Hz),3.07～3.03(m,1H),2.86(s,3H),2.63(t,1H,J＝8.8Hz),2.23(s,6H),2.01(s,3H),2.03(s,3H),1.94(s,1H),1.86～1.81(m,1H),1.73～1.70(m,1H),1.59～1.54(m,3H),1.47(s,3H),1.36～1.34(m,3H),1.32(s,3H),1.26～1.22(m,3H),1.15～1.13(m,6H),0.92(t,3H,J＝7.2Hz).

¹³C NMR(100MHz,CDCl₃):207.40,170.14,170.10,142.44,139.16,102.17,81.69,81.32,78.67,73.74,71.91,69.47,63.92,51.58,50.77,47.50,41.02,40.52,38.81,30.75,22.79,22.35,21.75,21.38,19.28,15.16,14.39,13.94,11.31.

HR-MS(ESI)(M+H)⁺m/z 612.3755, calculated: c₃₂H₅₄NO₁₀612.3742.

2.57g, 4.206mmol of Compound 5 were dissolved in 25ml of dry N, N-Dimethylformamide (DMF), the reaction was cooled to-20 ℃ in a low temperature cooling tank, and 0.336g, 8.412mmol of sodium hydride was slowly added thereto

The feeding speed is based on the air discharging speed, and is not suitable to be too fast. After reacting for 30min, slowly adding N, N-Carbonyl Diimidazole (CDI) into the reaction system, continuing to react for 2h, and then monitoring by LC-MS that no raw material exists in the reaction system, namely stopping the reaction. Adding a little water into the reaction system for quenching, adding 125ml of ethyl acetate for diluting, washing with 60ml of saturated saline for 6 times, separating liquid, drying an organic phase with anhydrous sodium sulfate, filtering and evaporating to dryness. Separating by column chromatography, soaking with petroleum ether, eluting with ethyl acetate, mixing the components, and evaporating to dryness to obtain white foamy solid 6(2.608g, 92.1%).

¹H NMR(400MHz,CDCl₃):δ8.07(s,1H),7.35(s,1H),7.05(s,1H),6.78(s,1H),5.67(dd,1H,J＝9.6Hz,J＝2.4Hz),4.70(t,1H,J＝8.0Hz),4.33(d,1H,J＝7.6Hz),4.109(d,1H,J＝8.8Hz),3.73(q,1H,J＝6.8Hz),3.49～3.45(m,1H),3.14(d,1H,J＝6.4Hz),3.01(t,3H,J＝6.0Hz),2.76(s,3H),2.62(t,1H,J＝8.8Hz),2.24(s,6H),2.02(s,3H),1.84～1.80(m,6H),1.72～1.60(m,3H),1.43～1.41(m,3H),1.35～1.34(m,3H),1.39(s,3H),1.22～1.19(m,3H),0.93(t,3H,J＝7.2Hz).

¹³C NMR(100MHz,CDCl₃):205.25,169.98,169.12,146.22,138.42,137.32,131.19,117.38,102.22,84.81,81.23,78.82,71.81,69.42,63.35,53.70,51.30,50.54,47.68,40.91,40.56,39.22,30.55,22.92,21.65,21.21,20.97,20.44,19.19,15.33,14.29,13.54,10.74.

HR-MS(ESI)(M+H)⁺m/z 706.3895, calculated: c₃₃H₅₈N₂O₁₄706.3883.

Dissolving 5.75g and 8.17mmol of compound 6 in 64ml of anhydrous DMF and 6ml of tetrahydrofuran, placing the reaction system in a low-temperature cooling tank, cooling to-20 ℃, adding 1.10g and 9.79mmol of potassium tert-butoxide into the reaction system, reacting for 30min, adding 2.83g and 8.99mmol of N-fluorobis-benzenesulfonamide (NISF) into the reaction system, reacting for 2h, and detecting by LC-MS that no raw material remains in the reaction, namely stopping the reaction. Adding a small amount of water into the reaction system for quenching, taking 250ml of ethyl acetate to dilute the reaction solution, washing with 120ml of saturated saline solution for 6 times, and separating the solution. The organic phase was dried over anhydrous sodium sulfate, filtered and spin dried. Column chromatography separation, petroleum ether infiltration, ethyl acetate elution, merging of components and spin drying to obtain a white foamy solid 7(5.49g, 93.2%).

¹H NMR(400MHz,CDCl₃)δ8.09(s,1H),7.37(s,1H),7.07(dd,J＝1.6,0.8Hz,1H),6.69(s,1H),5.54(dd,J＝9.3,2.9Hz,1H),4.74(dd,J＝10.6,7.6Hz,1H),4.37(d,J＝7.6Hz,1H),3.97(dd,J＝10.1,1.4Hz,1H),3.51–3.44(m,2H),2.67–2.60(m,4H),2.24(s,6H),2.07(s,3H),1.90(s,4H),1.80(dd,J＝7.3,4.1Hz,4H),1.78–1.73(m,4H),1.28(dd,J＝6.6,4.7Hz,4H),1.26–1.20(m,9H),1.17(d,J＝7.0Hz,3H),0.99(d,J＝7.5Hz,3H).

HR-MS(ESI)(M+H)⁺m/z 724.3766, calculation C₃₆H₅₄FN₃O₁₁723.3743.

11.58g, 16.41mmol of Compound 7 were dissolved in 150ml of anhydrous methanol, stirred overnight and spun dry to give Compound 8 as a white foam (7.83g, 76.2%).

HR-MS(ESI)(M+H)⁺m/z 682.3670,calcd for C₃₄H₅₂FN₃O₁₀681.3637.

Preparation example 2 Synthesis route of intermediate side chain a4

The Scheme 1 reagent and the reaction conditions are that a, di-tert-butyl dicarbonate, sodium bis (trimethylsilyl) amide and tetrahydrofuran are mixed at room temperature for 2 hours, and the mixture is 92.8 percent; b. nitrogen-3-bromopropyl-phthalimide, potassium carbonate, N, N-dimethylformamide, 90 ℃, 85.2% for 8 hours; c. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours, 97.3%; d. hydrazine hydrate, ethanol, 4 hours, 80 ℃, 82.7%;

10.02g, 69.50mmol of 3-aminoquinoline are dissolved in 150ml of dry tetrahydrofuran, the reaction system is placed in an ice water bath for cooling, 69ml of sodium bis (trimethylsilyl) amide (2mol/L tetrahydrofuran solution) is slowly added into the reaction system under the protection of argon, then 16.68g, 76.45mmol of di-tert-butyl dicarbonate is added, the reaction solution is gradually returned to room temperature, and TLC (petroleum ether: ethyl acetate 1:1) monitors that the reaction is complete, namely the reaction is stopped. Adding a little water into the reaction system to quench the reaction, spin-drying the reaction solution, diluting the residue with 450ml of ethyl acetate, washing the residue with 200ml of saturated sodium bicarbonate solution and saturated saline solution for 1 time respectively, separating the solution, and drying the organic phase with anhydrous sodium sulfate. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate at a ratio of 5:1, mixing the components, and spin-drying to obtain white cotton-like solid a 1.

12.85g, 52.66mmol of compound a1 is dissolved in 250ml of dry N, N-dimethylformamide, the reaction system is placed in an ice-water bath for cooling, then 4.21g, 105.32mmol of sodium hydride are slowly added, the reaction system is in a viscous state, after stirring for 30min, 28.23g, 105.32mmol of N-3-bromopropylphthalimide is slowly added to the reaction solution, after stirring for 15min at the constant temperature, the reaction solution is heated to 80 ℃ for overnight reaction, and the reaction is stopped as monitored by TLC (petroleum ether: ethyl acetate 1:1) for completion of the reaction. The reaction mixture was diluted with 750ml of ethyl acetate, washed 6 times with 350ml of saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 7:1 combined fractions and spin dried to give a white solid a2(20.44g, 91.3%).

3.26g, 7.56mmol of compound a2 was dissolved in 35ml of anhydrous dichloromethane, the reaction system was cooled in an ice-water bath, 17.24g, 11.24ml, 151.2mmol of trifluoroacetic acid were slowly added to the reaction system, the reaction system was gradually returned to room temperature, and after the reaction was sufficiently stirred for 8 hours, the completion of the reaction was monitored by TLC (petroleum ether: ethyl acetate ═ 1:1), i.e., the reaction was stopped. The reaction mixture was centrifuged to remove a large amount of remaining trifluoroacetic acid, and the residue was dissolved in 175ml of dichloromethane, washed with 80ml of saturated sodium bicarbonate solution and saturated brine each for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 10:1 combined fractions and spin dried to give a pale yellow solid a3(2.4g 96.1%).

0.42g of 1.27mmol of compound a3 was dispersed in 10ml of ethanol, 0.15ml of 2.54mmol of 80% hydrazine hydrate solution was slowly added to the reaction solution, and the reaction was heated to 80 ℃ for 2 hours, whereby it was observed that the starting material was dissolved in the solvent first and then a large amount of white solid was precipitated. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. Cooling the reaction liquid to room temperature, removing a large amount of ethanol by spinning, dissolving the residue in 25ml of dichloromethane, performing ultrasonic extraction for 15min, performing suction filtration (with diatomite as a filter aid), adding 25ml of dichloromethane into the filter cake again, performing ultrasonic extraction for 15min, combining the two filtrates, performing spin drying to obtain a light yellow oily liquid a4, and directly putting the light yellow oily liquid a4 into the next reaction without separation.

Preparation example 2 Synthesis route of intermediate side chain b2

Scheme 2 reagents and conditions a, benzoyl chloride, triethylamine, room temperature, 4 hours, 92.5%; b. hydrazine hydrate, ethanol, 80 ℃,4 hours, 82.3%.

1.57mmol, 0.52g of compound a2 was dissolved in 10ml of pyridine, the reaction system was placed in an ice-water bath, 0.32, 2.26mmol and 0.27ml of benzoyl chloride were slowly added to the reaction system, and after continuing the reaction for 4 hours, the completion of the reaction was monitored by TLC (petroleum ether: ethyl acetate ═ 1:1), and the reaction was stopped. The reaction mixture was spin-dried, the residue was dissolved in 25ml of dichloromethane, washed with 10ml of a 5% potassium dihydrogen phosphate solution and saturated brine for 1 time, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 7:1 combined the fractions and spin dried to give b1 as a white powder (0.638g, 87.2%). 0.63g of 1.45mmol of compound b2 was dissolved in 15ml of absolute ethanol, 0.23ml of 3.62mmol of 80% hydrazine hydrate solution was slowly added to the reaction solution, and the reaction was heated to 80 ℃ for 2 hours, whereby it was observed that the starting material was dissolved in the solvent first and then a large amount of white solid was precipitated. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. Filtering the reaction solution, directly carrying out column chromatography separation on the filtrate, soaking the filtrate in dichloromethane, and carrying out dichloromethane: methanol 100: 1 to obtain a byproduct compound 28 (b); dichloromethane: methanol 20:1 elution, combined fractions and spin dried to give b2 as a white oily liquid (0.17g, 42.7%).

Preparation examples 3-5 synthetic routes to intermediate side chains c3-e3

Scheme 3 reagents and conditions: a. acetic anhydride (c1), phenylpropionyl chloride (d1), cinnamoyl chloride (e1), triethylamine, room temperature, 4 hours; b. sodium hydride, N-tert-butoxycarbonyl-3-amino-propyl bromide, N, N-dimethylformamide, at 90 ℃ for 8 hours; c. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours.

Dissolving 3-aminoquinoline in anhydrous dichloromethane, cooling in an ice-water bath, slowly adding 1.5eq of dried triethylamine, fully stirring, slowly adding 1.1eq of acetic anhydride (c), phenylpropionyl chloride (d) and cinnamoyl chloride (e), reacting for 2 hours, and detecting complete reaction by TLC (petroleum ether: ethyl acetate 1:1), namely stopping the reaction. The reaction mixture was diluted with 350ml of dichloromethane, washed 2 times with 5% potassium dihydrogen phosphate solution and saturated brine, separated, the organic phase was dried over anhydrous sodium sulfate, and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate 10:1, combining the components, and spin-drying to obtain c1-e 1. Compound c1(d1, e1) was dissolved in anhydrous N, N-dimethylformamide, cooled in an ice-water bath, and 2eq of sodium hydride was slowly added thereto, with attention paid to the rate of gas evolution in the reaction system, and after stirring for 30 minutes, 3eq of N-Boc-3-aminopropyl bromide was added, and the reaction was heated at 90 ℃ for 8 hours, and the reaction was stopped when the reaction was completed as checked by TLC (petroleum ether: ethyl acetate ═ 1: 1). The reaction system was cooled to room temperature, diluted with dichloromethane, washed with saturated brine 6 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate 7:1, combining the components, and spin-drying to obtain c2-e 2.

Dissolving c2(d2, e2) in anhydrous dichloromethane, cooling the reaction system under the condition of ice-water bath, slowly adding 17mL of trifluoroacetic acid into the reaction solution, fully stirring, gradually returning the reaction solution to room temperature, continuing the reaction for 8 hours, and detecting the completion of the reaction by TLC (dichloromethane: methanol: 10:1), namely stopping the reaction. Removing a large amount of trifluoroacetic acid from the reaction solution by spinning, and directly carrying out column chromatography separation. Dichloromethane infiltration, dichloromethane: eluting with methanol 10:1, mixing the components, and spin-drying to obtain c3-e 3.

Preparation 6 Synthesis of intermediate side chain f4

Scheme 4 reagent and conditions a benzyl bromide, potassium carbonate, room temperature, 4 hours, 83%; b. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours, 95%; c.N- (3-bromopropyl-) phthalimide, potassium carbonate, N, N-dimethylformamide, 90 ℃, argon protection, 8 hours and 78 percent; d. hydrazine hydrate, ethanol, 4 hours, 80 ℃, 82%.

12.2g and 0.05mol of compound a1 were dissolved in 120mL of anhydrous N, N-dimethylformamide, the mixture was cooled in an ice-water bath, 4g and 0.1mol of 60% sodium hydride were added to the reaction system, and after stirring thoroughly for 30min, 7.1mL, 10.265g and 0.06mol of benzyl bromide were added to the reaction mixture, and the mixture was heated to 90 ℃ overnight and the reaction was stopped by monitoring completion of the reaction by TLC (petroleum ether: ethyl acetate: 1). The reaction mixture was diluted with 600mL of dichloromethane, washed with 300mL of saturated brine 6 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether: ethyl acetate 10:1 elution, combine fractions and spin dry to give f1 as a pale yellow oil (16.2g, 83.6%).

6.68g, 20mmol of the compound f1 was dissolved in 70mL of anhydrous dichloromethane, the reaction system was cooled in an ice water bath, 35mL of trifluoroacetic acid was slowly added to the reaction system, the reaction system was gradually returned to room temperature, and after the reaction was sufficiently stirred for 8 hours, the reaction was monitored by TLC (petroleum ether: ethyl acetate ═ 1:1) for completion, i.e., the reaction was stopped. The reaction mixture was centrifuged to remove a large amount of remaining trifluoroacetic acid, and the residue was dissolved in 175ml of dichloromethane, washed with 80ml of saturated sodium bicarbonate solution and saturated brine each for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 10:1 combined fractions and spin dried to give f2 as a pale yellow solid (23.42g, 95.3%).

4.68g, 20mmol of compound f2 was dissolved in 75ml of dry DMF, the reaction system was cooled in an ice-water bath, 16.086g, 60mmol of 60% sodium hydride was slowly added thereto, the reaction system was in a viscous state, after stirring for 30min, 16.07g, 60mmol of N-3-bromopropylphthalimide was slowly added to the reaction solution under argon protection, the reaction was continued for 8h, and the completion of the reaction was monitored by TLC (petroleum ether: ethyl acetate 1:1), i.e., the reaction was stopped. The reaction mixture was diluted with 750ml of ethyl acetate, washed 6 times with 350ml of saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: ethyl acetate 7:1 elution, combined fractions and spin dried to give f3 as a white solid (6.44g, 84.3%).

4.21g, 10mmol of compound f3 were dispersed in 50mL of ethanol, 0.5mL of 85% hydrazine hydrate solution was slowly added to the reaction solution, and the reaction was heated to 80 ℃ for 2h, and it was observed that the starting material was dissolved in the solvent first and then a large amount of white solid was precipitated. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. Cooling the reaction liquid to room temperature, removing a large amount of ethanol by spinning, dissolving the residue in 25ml of dichloromethane, performing ultrasonic extraction for 15min, performing suction filtration (with diatomite as a filter aid), adding 25ml of dichloromethane into the filter cake again, performing ultrasonic extraction for 15min, combining the two filtrates, performing spin drying to obtain a light yellow oily liquid f4, and directly putting the light yellow oily liquid into the next reaction without separation. Preparation example 7 Synthesis of intermediate side chain g4

Scheme 5 reagents and conditions: a. di-tert-butyl dicarbonate, sodium bis (trimethylsilyl) amide, tetrahydrofuran, room temperature, 2 hours, 91.3%; b. nitrogen-3-bromopropyl-phthalimide, potassium carbonate, N, N-dimethylformamide, 90 ℃, 85.2% for 8 hours; c. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours, 97.3%; d. hydrazine hydrate, ethanol, 4 hours, 80 ℃, 82.7%.

5.02g, 39.75mmol of 4-aminoquinoline was dissolved in 75ml of dry tetrahydrofuran, the reaction system was cooled in an ice-water bath, 35ml of NHMDS (2mol/L tetrahydrofuran solution) was slowly added to the reaction system under the protection of argon, then 8.84g, 38.27mmol of Boc anhydride was added, the reaction solution was gradually returned to room temperature, and the reaction was monitored by TLC (petroleum ether: ethyl acetate ═ 1:1) for completion, i.e., the reaction was stopped. Adding a little water into the reaction system to quench the reaction, spin-drying the reaction solution, diluting the residue with 450ml of ethyl acetate, washing the residue with 200ml of saturated sodium bicarbonate solution and saturated saline solution for 1 time respectively, separating the solution, and drying the organic phase with anhydrous sodium sulfate. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 5:1 combined fractions and spin dried to give g1 as a white cotton solid (6.97g, 91.3%).

6.43g, 26.33mmol of compound g1 was dissolved in 100ml dry DMF, the reaction system was cooled in an ice water bath, 2.15g, 52.66mmol of sodium hydride was slowly added thereto, the reaction system was in a viscous state, after stirring for 30min, 14.11g, 52.66mmol of N-3-bromopropylphthalimide was slowly added to the reaction solution, after stirring for 15min at constant temperature, the reaction was heated to 80 ℃ overnight, and the reaction was stopped by TLC (petroleum ether: ethyl acetate 1:1) monitoring completion of the reaction. The reaction mixture was diluted with 750ml of ethyl acetate, washed 6 times with 350ml of saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 7:1 combined fractions and spin dried to yield g2(10.14g, 89.37%) as a pale yellow liquid.

1.63g, 3.78mmol of compound g2 was dissolved in 25mL of anhydrous dichloromethane, the reaction system was cooled in an ice-water bath, 12mL of trifluoroacetic acid was slowly added to the reaction system, the reaction system was gradually returned to room temperature, and after the reaction was sufficiently stirred for 8 hours, the reaction was monitored by TLC (petroleum ether: ethyl acetate 1:1) for completion, i.e., the reaction was stopped. The reaction mixture was centrifuged to remove a large amount of remaining trifluoroacetic acid, and the residue was dissolved in 175ml of dichloromethane, washed with 80ml of saturated sodium bicarbonate solution and saturated brine each for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: elution with ethyl acetate 10:1 combined fractions and spin dried to yield g3 as a pale yellow solid (1.25g 96.7%).

0.84g, 2.54mmol of compound g3 was dispersed in 15ml ethanol, 0.3ml of 80% hydrazine hydrate solution was slowly added to the reaction solution, and the reaction was heated to 80 ℃ for 2 hours, and it was observed that the starting material was dissolved in the solvent first and then a large amount of white solid was precipitated. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. Cooling the reaction liquid to room temperature, removing a large amount of ethanol by spinning, dissolving the residue in 25ml of dichloromethane, performing ultrasonic extraction for 15min, performing suction filtration (with diatomite as a filter aid), adding 50ml of dichloromethane into the filter cake again, performing ultrasonic extraction for 15min, combining the two filtrates, performing spin drying to obtain a light yellow oily liquid g4, and directly putting the light yellow oily liquid into the next reaction without separation.

Preparation example 8 Synthesis of intermediate side chain h4-j4

Scheme 6 reagents and conditions: a. acetic anhydride (c1), benzoyl chloride (d1), cinnamoyl chloride (e1), triethylamine, room temperature, 4 hours; b. sodium hydride, N-tert-butoxycarbonyl-3-amino-propyl bromide, N, N-dimethylformamide, at 90 ℃ for 8 hours; c. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours.

Dissolving 4-aminoquinoline in anhydrous dichloromethane, cooling in an ice-water bath, slowly adding 1.5eq of dry triethylamine, fully stirring, slowly adding 1.2eq of acetic anhydride (h), benzoyl chloride (i) and cinnamoyl chloride (j) respectively, reacting for 2h, and detecting complete reaction by TLC (petroleum ether: ethyl acetate 1:1), namely stopping the reaction. The reaction mixture was diluted with dichloromethane, washed 2 times with 5% potassium dihydrogen phosphate solution and saturated brine, separated, the organic phase was dried over anhydrous sodium sulfate, and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate 10:1, mixing the components, and spin-drying to obtain white cotton-like solid h1-j 1.

The compound h1-j1 was dissolved in 80ml of anhydrous N, N-dimethylformamide, cooled in an ice-water bath, and 2eq of sodium hydride was slowly added thereto, with attention paid to the rate of gas evolution in the reaction system, and after stirring for 30 minutes, 3eq of N-t-butoxycarbonyl-3-aminopropyl bromide was added thereto, and the mixture was heated to 90 ℃ for reaction for 8 hours, and the reaction was stopped when the reaction was completed as detected by TLC (petroleum ether: ethyl acetate 1: 1). The reaction system was cooled to room temperature, diluted with dichloromethane, washed with saturated brine 6 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate 7:1, mixing the components, and spin-drying to obtain a light yellow powdery solid h2-j 2.

Respectively dissolving the compound h2-j2 in 50mL of anhydrous dichloromethane, cooling the reaction system under the condition of ice-water bath, slowly adding 25mL of trifluoroacetic acid into the reaction solution, fully stirring, gradually returning the reaction solution to room temperature, continuously reacting for 8 hours, and detecting the completion of the reaction by TLC (dichloromethane: methanol: 10:1), namely stopping the reaction. Removing a large amount of trifluoroacetic acid from the reaction solution by spinning, and directly carrying out column chromatography separation. Dichloromethane infiltration, dichloromethane: eluting with methanol 10:1, mixing the components, and spin-drying to obtain light brown foam solid h3-j3

Preparation example 9 Synthesis of intermediate side chain k2

Scheme 7 reagent and conditions are a, 1-bromobutane, potassium carbonate, N, N-dimethylformamide, 80 ℃,8 hours, 80%; b. hydrazine hydrate, ethanol, 80 ℃,4 hours, 82%.

15mmol, 4.97g of compound g3 was dissolved in 50mL of dry N, N-dimethylformamide, 1.2g, 30mmol of 60% sodium hydride was added, and after stirring well for 30min, 75mmol, 12.26g of 1-bromobutane was added, and the reaction was heated to 90 ℃ overnight, and the reaction was stopped upon completion of TLC (dichloromethane: methanol ═ 10: 1). The reaction mixture was returned to room temperature, diluted with 250mL of dichloromethane, washed with 120mL of saturated brine 6 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. And (4) spin-drying to obtain a compound k1, and directly putting the compound k1 into the next reaction.

6.97g, 0.018mmol of compound k1 was dispersed in 100ml of ethanol, 0.5ml of 80% hydrazine hydrate solution was slowly added to the reaction solution, and the reaction was heated to 80 ℃ for 2 hours, whereby it was observed that the starting material was dissolved in the solvent first and then a large amount of white solid was precipitated. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. Cooling the reaction liquid to room temperature, removing a large amount of ethanol by spinning, dissolving the residue in 150ml of dichloromethane, performing ultrasonic extraction for 15 minutes, performing suction filtration (with diatomite as a filter aid), adding 150ml of dichloromethane into the filter cake again, performing ultrasonic extraction for 15 minutes, combining the two filtrates, performing spin drying to obtain a light yellow oily liquid k2, and directly putting the light yellow oily liquid k2 into the next reaction without separation.

Preparation example 10 Synthesis of the intermediate side chain l4-m4

Scheme 8 reagents and conditions: a. methyl iodide (l), benzyl bromide (m), sodium hydride, N, N-dimethylformamide at room temperature for 2 hours; b. trifluoroacetic acid, dichloromethane, 0 ℃,8 hours; c.N- (3-bromopropyl) phthalimide, potassium carbonate, N, N-dimethylformamide, 80 ℃,8 h; d. hydrazine hydrate, ethanol, 80 ℃,4 hours.

Dissolving compound g1 in dry N, N-dimethylformamide, slowly adding 1.2eq of sodium hydride under the condition of ice-water bath, fully stirring for 30 minutes, then respectively adding 1.2eq of methyl iodide (l) and benzyl bromide (m), gradually returning the reaction system to the room temperature, continuing the reaction for 2 hours, and monitoring the completion of the reaction by TLC (petroleum ether: ethyl acetate 1:1), namely stopping the reaction. The reaction mixture was diluted with 300mL of dichloromethane, washed 6 times with 150mL of saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: ethyl acetate 7:1, eluting, combining the components, and spin-drying to obtain yellow oily liquid l1 and m 1.

Respectively dissolving the compounds l1 and m1 in anhydrous dichloromethane, placing the reaction system in an ice-water bath for cooling, then slowly adding trifluoroacetic acid into the reaction system, gradually returning the reaction system to the room temperature, fully stirring for reaction for 8 hours, and monitoring the reaction completion by TLC (dichloromethane: methanol ═ 10:1), namely stopping the reaction. The reaction solution was spun off a large amount of remaining trifluoroacetic acid, and the residue was dissolved in dichloromethane, separated by column chromatography, saturated with dichloromethane, dichloromethane: eluting with 20:1 methanol, mixing the components, and spin-drying to obtain light yellow solid l2 and m 2.

Respectively dissolving compounds l2 and m2 in dry N, N-dimethylformamide, placing the reaction system in an ice-water bath for cooling, then slowly adding 2eq of sodium hydride, keeping the reaction system in a viscous state, stirring for 30 minutes, then slowly adding 3eq of N-3-bromopropylphthalimide into the reaction solution, keeping the temperature and stirring for 15 minutes, heating to 80 ℃ for overnight reaction, and monitoring the completion of the reaction by TLC (dichloromethane: methanol ═ 10:1), namely stopping the reaction. The reaction solution was diluted with ethyl acetate, washed with saturated brine 6 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography separation, petroleum ether infiltration, petroleum ether: eluting with ethyl acetate 7:1, combining the components, and spin-drying to obtain white solids l3 and m 3.

Respectively dissolving the compounds l3 and m3 in ethanol, slowly adding 2eq of 80% hydrazine hydrate solution into the reaction solution, heating to 80 ℃ and reacting for 2 hours, and observing that the raw materials are firstly dissolved in the solvent and then a large amount of white solid is separated out. TLC (dichloromethane: methanol ═ 10:1) monitored the reaction was complete, i.e. stopped. And cooling the reaction liquid to room temperature, removing a large amount of ethanol by spinning, dissolving the residue in dichloromethane, performing ultrasonic extraction for 15 minutes, performing suction filtration (using diatomite as a filter aid), adding 150ml of dichloromethane into the filter cake again, performing ultrasonic extraction for 15 minutes, combining the two filtrates, and performing spin drying to obtain light yellow oily liquids l4 and m4, wherein the light yellow oily liquids are directly put into the next reaction without separation.

EXAMPLE 1 preparation of Compound S1

Synthetic route to compound S1:

0.228g, 1.13mmol of compound a4 was dissolved in 2.7ml of acetonitrile and 0.3ml of water, 0.45mol of compound 8 was added to the reaction system, and the mixture was heated to 90 ℃ and refluxed for 8 hours, and the reaction was monitored by TLC (dichloromethane: methanol ═ 10:1) for completion of the reaction, i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 6ml of dichloromethane, washed with 3ml of 5% potassium dihydrogen phosphate and saturated brine each for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S1(0.574g, 42.3%).

¹H NMR(500MHz,CDCl₃)δ8.48(d,J＝2.0Hz,1H),7.91(d,J＝7.8Hz,1H),7.61(d,J＝7.6Hz,1H),7.42～7.35(m,2H),6.99(s,1H),4.89(d,J＝9.6Hz,1H),4.77(s,1H),4.30(d,J＝7.1Hz,1H),4.05(d,J＝10.5Hz,1H),3.81(dd,J＝15.6,6.8Hz,1H),3.76～3.68(m,1H),3.56～3.49(m,2H),3.47(s,1H),3.28(dd,J＝11.6,5.6Hz,1H),3.25～3.17(m,2H),3.14(q,J＝6.5Hz,1H),2.64(d,J＝16.8Hz,2H),2.55(s,4H),2.48(t,J＝9.6Hz,1H),2.29(s,7H),2.13～2.03(m,1H),1.99～1.93(m,2H),1.89(d,J＝12.9Hz,1H),1.78(d,J＝21.4Hz,3H),1.65(dd,J＝27.1,9.8Hz,3H),1.51(s,3H),1.34(s,3H),1.30(d,J＝6.9Hz,3H),1.24(d,J＝6.0Hz,6H),1.20(d,J＝6.9Hz,4H),1.05(d,J＝6.8Hz,3H),0.85(t,J＝7.3Hz,4H).

¹³C NMR(125MHz,CDCl₃)δ217.28,202.88,202.65,166.54,166.31,157.48,143.64,141.90,141.79,129.06,128.93,126.87,126.03,124.64,109.47,104.17,98.58,96.53,82.33,80.75,78.73,70.45,69.71,66.00,61.36,53.54,49.41,44.65 41.82,40.93,40.24,39.65,39.34,28.27,26.30,25.47,25.29,22.27,21.30,19.86,15.18,14.70,10.51.

HR-MS(ESI)(M+H)⁺m/z 815.4562, calculation C₄₃H₆₃FN₄O₁₀814.4528.

EXAMPLE 2 preparation of Compound S2

Synthetic route to compound S2:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 2.5mmol and 0.710g of compound b2 were added to the reaction solution, and the mixture was heated at 90 ℃ for 8 hours, followed by completion of the reaction by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S2(0.634g, 39.8%).

¹H NMR(400MHz,CDCl₃)δ8.61(d,J＝2.4Hz,1H),7.99(d,J＝8.4Hz,1H),7.95(d,J＝2.1Hz,1H),7.74～7.68(m,2H),7.68～7.62(m,1H),7.51(ddd,J＝9.7,6.3,2.3Hz,2H),7.32～7.27(m,2H),7.18～7.13(m,1H),7.11(t,J＝7.2Hz,2H),4.87(d,J＝10.3Hz,1H),4.36(d,J＝7.1Hz,1H),4.21(t,J＝6.0Hz,1H),4.10(d,J＝7.0Hz,2H),4.02(dd,J＝10.7,1.7Hz,1H),3.76(dd,J＝8.6,5.4Hz,1H),3.71～3.62(m,1H),3.57(dd,J＝9.6,6.0Hz,1H),3.50(ddd,J＝10.4,7.1,3.4Hz,1H),3.14–3.01(m,2H),2.62(s,6H),2.43(s,3H),1.97～1.88(m,3H),1.81(d,J＝14.3Hz,1H),1.73(d,J＝21.4Hz,4H),1.47(s,4H),1.35～1.21(m,17H),1.17(d,J＝6.8Hz,3H),0.98(d,J＝6.9Hz,3H),0.91～0.84(m,4H),0.79(t,J＝7.4Hz,3H).

¹³C NMR(100MHz,CDCl₃)δ216.48,202.82,202.43,170.66,157.19,150.34,146.16,136.87,135.65,133.00,131.00,129.67,129.27,128.91,128.82,128.15,127.96,127.84,127.33,103.33,78.69,78.53,69.94,68.58,68.27,66.42,61.40,49.28,48.13,44.68,41.77,40.79,40.40,39.60,39.38,38.83,30.46,30.22,29.03,25.39,25.16,23.85,22.28,21.06,19.77,18.02,15.16,14.88,14.17,13.86,10.37.

HR-MS(ESI)(M+H)⁺m/z 932.4956, calculation C₅₁H₆₉FN₄O₁₁932.4012

EXAMPLE 3 preparation of Compound S3

Synthetic route to compound S3:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.687g of compound c3 was added to the reaction solution, and the mixture was heated at 90 ℃ for 8 hours, followed by completion of the reaction by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S3(0.582g, 40.3%).

¹H NMR(400MHz,CDCl₃)δ8.72(d,J＝2.5Hz,1H),8.16(d,J＝8.4Hz,1H),7.97(dd,J＝2.5,0.8Hz,1H),7.86(dd,J＝8.3,1.4Hz,1H),7.80(ddd,J＝8.5,6.9,1.4Hz,1H),7.65(ddd,J＝8.1,6.9,1.2Hz,1H),6.74(s,1H),5.77(t,J＝6.3Hz,1H),5.56(s,1H),4.36(d,J＝7.1Hz,1H),4.04(d,J＝8.9Hz,1H),3.86(td,J＝6.4,3.6Hz,2H),3.54(d,J＝31.2Hz,2H),3.42(dd,J＝10.1,7.1Hz,1H),3.21(q,J＝6.2Hz,2H),2.81(s,2H),2.67(s,6H),1.90(s,3H),1.75(d,J＝21.8Hz,3H),1.68(s,3H),1.65～1.59(m,3H),1.42–1.36(m,1H),1.27(d,J＝6.1Hz,3H),1.26～1.23(m,6H),1.19(d,J＝6.7Hz,3H),0.96(t,J＝7.5Hz,3H).

¹³CNMR(100MHz,CDCl₃)δ216.48,202.82,202.47,170.65,167.88,166.37,166.14,150.33,136.86,132.99,131.00,129.66,129.27,128.90,128.81,128.15,127.96,127.32,103.32,82.15,80.69,78.69,78.52,69.93,68.57,68.26,66.41,61.40,49.28,48.12,44.67,41.77,40.78,39.59,39.37,38.83,30.46,30.21,29.03,26.25,25.16,23.84,23.09,22.28,21.05,19.77,18.02,15.16,14.87,14.16,13.86,10.36.

HR-MS(ESI)(M+H)⁺m/z 872.5672, calculation C₄₆H₆₇FN₄O₁₁871.4833.

EXAMPLE 4 preparation of Compound S4

Synthetic route to compound S4:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 1.05g of compound d3 was added to the reaction solution, the mixture was heated at 90 ℃ for 8 hours, and the reaction was stopped by TLC (dichloromethane: methanol ═ 10:1) after completion of the reaction. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S4(0.692g, 41.6%).

¹H NMR(500MHz,CDCl₃)δ8.52(s,1H),8.15(d,J＝8.4Hz,1H),7.81(t,J＝7.5Hz,1H),7.76(d,J＝8.1Hz,1H),7.64(t,J＝7.4Hz,1H),7.47(s,1H),7.29(d,J＝13.6Hz,2H),7.07(d,J＝5.8Hz,2H),6.81(s,1H),5.83(s,1H),5.60(s,1H),4.38(d,J＝7.0Hz,1H),4.07(d,J＝9.3Hz,1H),3.84(s,2H),3.64～3.50(m,3H),3.42～3.37(m,1H),3.20～3.10(m,4H),2.96(t,J＝6.9Hz,3H),2.82(s,3H),2.59(s,6H),2.39(t,J＝6.8Hz,2H),1.93(s,4H),1.79(d,J＝21.8Hz,5H),1.73(s,5H),1.31(s,3H),1.29(d,J＝6.3Hz,9H),1.23(d,J＝5.3Hz,3H),1.01(t,J＝7.1Hz,3H),0.94–0.85(m,1H).

¹³C NMR(100MHz,CDCl₃)δ216.42,209.85,206.42,173.12,166.22,165.99,155.02,154.26,150.09,147.29,142.34,140.62,136.86,135.08,134.54,130.58,130.00,129.49,128.65,128.62,127.98,127.88,127.77,126.54,102.36，85.84,83.56，79.29,70.32,68.91,65.87,49.70,46.43,41.14,40.54,37.07,36.34,31.84,30.38,29.40,27.29,27.44,25.62,25.11,24.04,23.81,22.53,20.99,20.48,20.06,19.51,14.21,13.15,10.78.

HR-MS(ESI)(M+H)⁺m/z 961.5388, calculation C₅₃H₇₃FN₄O₁₁960.5362

EXAMPLE 5 preparation of Compound S5

Synthetic route to compound S5:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 1.04g of compound e3 was added to the reaction solution, the mixture was heated at 90 ℃ for 8 hours, and the reaction was stopped by TLC (dichloromethane: methanol ═ 10:1) after completion of the reaction. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S5(0.682g, 40.9%).

¹H NMR(400MHz,CDCl₃)δ8.90(d,J＝2.5Hz,1H),8.27～8.17(m,2H),7.94(d,J＝8.1Hz,1H),7.82(t,J＝7.7Hz,1H),7.76(d,J＝15.4Hz,1H),7.66(t,J＝7.5Hz,1H),7.29(d,J＝4.2Hz,5H),6.27(d,J＝15.9Hz,1H),4.84(dd,J＝10.4,2.6Hz,1H),4.41(d,J＝7.1Hz,1H),4.06(t,J＝9.3Hz,3H),3.83(ddd,J＝14.6,9.4,5.3Hz,2H),3.66(ddt,J＝19.0,10.6,5.2Hz,3H),3.60～3.52(m,2H),3.48(dd,J＝10.1,7.1Hz,2H),3.11(q,J＝7.8,7.0Hz,3H),2.69(s,6H),2.50(s,2H),2.01～1.91(m,3H),1.79(s,4H),1.50(s,3H),1.44(s,1H),1.35～1.29(m,10H),1.22(d,J＝6.8Hz,3H),1.03(d,J＝6.9Hz,3H),0.81(t,J＝7.3Hz,3H).

¹³C NMR(100MHz,CDCl₃)δ216.45,202.89,202.52,173.28,166.01,165.83,155.26,154.95,150.68,147.18,143.32,140.67,135.54,134.54,129.81,129.48,128.76,128.36,128.09,127.61,127.88,127.77,126.54,118.27,103.29,83.56，82.19,80.76,78.52,69.95,68.54,66.63,61.51,49.27,47.75,44.66,42.01,40.77,40.46,39.62,39.35,30.34,26.39,25.41,25.19,22.23,21.05,14.80,13.89,10.38.

HR-MS(ESI)(M+H)⁺m/z 958.5162, calculation C₅₃H₇₁FN₄O₁₁957.5147

EXAMPLE 6 preparation of Compound S6

Synthetic route to compound S6:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.87g of compound f4 was added to the reaction solution, and the mixture was heated at 90 ℃ for 8 hours, followed by completion of the reaction by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S6(0.652g, 40.7%).

¹H NMR(400MHz,CDCl₃):δ8.09(d,J＝2.4Hz,1H),8.05(s,1H),7.92(s,1H),7.81～7.77(m,2H),7.71～7.65(m,1H),7.59～7.54(m,2H),7.47(d,1H,J＝8.0Hz),4.83(m,1H),4.72(t,J＝9.6Hz,1H),4.33(d,J＝7.6Hz,1H),4.13(d,J＝7.6Hz,1H),3.73～3.68(m,3H),3.57(d,J＝7.2Hz,1H),3.12～3.01(m,6H),2.42(d,J＝8.0Hz,3H),2.34(m,2H),2.25(s,2H),1.84(d,J＝21.4Hz,3H),1.64～1.59(m,3H),1.42(s,3H),1.28～1.25(m,6H),1.17(s,3H),1.11(t,J＝6.4Hz,3H),0.96(t,J＝8.0Hz,3H).

¹³C NMR(100MHz,CDCl₃):δ217.47,204.05,202.96,174.12,169.67，165.03,156.16，147.24,141.77，141.24，141.09，137.76,129.30,128.67,128.62,127.00,126.57,126.54,126.20,124.74,121.61,103.72,87.43,80.81,79.07,78.05,70.26,69.51,65.89,62.86,53.40,50.87,50.82,49.32,47.55,44.68,40.19,39.56,39.46,29.66,28.26,22.01,21.14,19.59,18.01,15.81,14.34,14.08,13.79,12.69,10.34.

HR-MS(ESI)(M+H)⁺m/z 919.5201, calculation C₅₁H₇₁FN₄O₁₀918.5190.

EXAMPLE 7 preparation of Compound S7

Synthetic route to compound S7:

0.684g, 3.39mmol of compound g4 was dissolved in 9ml acetonitrile and 1ml water, 1.35mol of compound 8 and 0.975g were added to the reaction system, and the reaction was refluxed at 90 ℃ for 8 hours, and the completion of the reaction was monitored by TLC (dichloromethane: methanol ═ 10:1), that is, the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 6ml of dichloromethane, washed with 3ml of 5% potassium dihydrogen phosphate and saturated brine each for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain S7(0.834g, 41.3%) as a dark yellow foamy solid.

¹H NMR(400MHz,CDCl₃)δ8.19(d,J＝9.9Hz,1H),8.15(s,1H),6.94(s,1H),4.87(d,J＝11.4Hz,1H),4.05(d,J＝10.7Hz,1H),3.92～3.83(m,1H),3.48(s,9H),3.26～3.20(m,1H),3.19～3.12(m,1H),2.68(d,J＝9.8Hz,1H),2.53(s,4H),2.31(s,6),2.08(s,3H),1.93(s,1H),1.90(s,1H),1.78(d,J＝21.3Hz,3H),1.68(s,1H),1.58(d,J＝13.3Hz,1H),1.53(s,3H),1.34(s,3H),1.31(d,J＝7.5Hz,3H),1.24(d,J＝7.6Hz,6H),1.06(s,3H),0.85(s,3H).

¹³C NMR(100MHz,CDCl₃)δ217.47,204.05,202.96,177.11,166.42,158.06,152.59,147.17,143.82,129.33,128.82,125.92,125.70,104.21,97.79,82.80,80.69,78.73,77.35,70.34,69.48,65.93,61.76,49.36,44.88,41.82,40.88,40.57,40.15,39.81,39.44,28.60,25.98,22.85,22.28,21.22,19.84,17.96,15.19,14.71,13.95,10.51.

HR-MS(ESI)(M+H)⁺m/z 815.4562, calculation C₄₃H₆₃FN₄O₁₀814.4528.

EXAMPLE 8 preparation of Compound S8

Synthetic route to compound S8:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.687g of compound h4 was added to the reaction solution, the mixture was heated at 90 ℃ for 8 hours, and the reaction was stopped by TLC (dichloromethane: methanol ═ 10:1) after completion of the reaction. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S8(0.563g, 39.6%).

¹H NMR(400MHz,CDCl₃)δ9.04(dd,J＝13.3,4.5Hz,1H),8.23(t,J＝8.2Hz,1H),7.88(t,J＝7.5Hz,1H),7.82(t,J＝7.7Hz,1H),7.67(q,J＝7.2Hz,1H),7.49(dd,J＝23.1,4.5Hz,1H),4.80(d,J＝10.0Hz,1H),4.40(d,J＝6.2Hz,1H),4.09(t,J＝11.8Hz,1H),3.76(d,J＝14.7Hz,1H),3.67～3.53(m,3H),3.40(d,J＝5.6Hz,1H),3.15～3.07(m,1H),2.86(s,1H),2.54(s,6H),2.52(s,4H),2.39(s,1H),1.92(dd,J＝22.5,7.3Hz,3H),1.83(d,J＝3.7Hz,3H),1.78(d,J＝4.9Hz,4H),1.50(s,3H),1.37(s,2H),1.35～1.29(m,10H),1.21(d,J＝6.7Hz,2H),1.04～0.98(m,3H),0.85(t,J＝7.4Hz,3H),0.77(t,J＝7.4Hz,3H).

¹³C NMR(100MHz,CDCl₃)δ216.60,202.75,202.35,169.96,166.38,157.19,151.26,150.23,147.35,130.42,128.23,125.88,122.61,121.47,103.72,98.93,96.88,82.22,80.84,78.47,70.17,69.05,66.43,61.25,49.32,46.71,46.40,44.64,41.38,40.46,39.67,39.28,29.55,26.74,25.22,22.59,22.20,21.17,19.85,18.00,15.16,14.82,13.89,10.47.

HR-MS(ESI)(M+H)⁺m/z 872.5672, calculation C₄₆H₆₇FN₄O₁₁871.4833

EXAMPLE 9 preparation of Compound S9

Synthetic route to compound S9:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 2.5mmol, 0.710g of compound i4 was added to the reaction solution, and the mixture was heated at 90 ℃ for 8 hours, followed by completion of the reaction by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S9(0.644g, 40.8%).

¹H NMR(400MHz,CDCl₃)δ8.74(d,J＝11.2Hz,1H),8.10(d,J＝7.6Hz,1H),7.99(d,J＝8.3Hz,1H),7.62(t,J＝7.7Hz,1H),7.21(d,J＝11.2Hz,3H),7.12(d,J＝6.4Hz,2H),7.05–6.95(m,2H),5.34(t,J＝5.0Hz,1H),4.81(d,J＝10.1Hz,1H),4.37～4.28(m,1H),4.08～4.00(m,1H),3.78(d,J＝13.7Hz,1H),3.63～3.58(m,1H),3.52(dq,J＝14.7,7.9,7.1Hz,3H),3.37(d,J＝8.3Hz,2H),3.25(t,J＝8.8Hz,1H),3.12～3.04(m,1H),2.60(d,J＝11.8Hz,2H),2.50(s,2H),2.37(s,7H),2.28(s,2H),2.21(t,J＝7.6Hz,2H),2.08(t,J＝9.6Hz,2H),2.05～1.97(m,5H),1.79(s,3H),1.63(p,J＝7.5Hz,2H),1.46(s,3H),1.30～1.26(m,19H),1.16(d,J＝6.8Hz,3H),0.97(d,J＝6.9Hz,3H),0.86(q,J＝8.7,7.5Hz,3H).

¹³C NMR(100MHz,CDCl₃)δ216.58,202.46,202.15,175.66,171.26,166.35,166.12,150.84,149.93,135.70,130.32,130.02,127.77,125.63,122.95,104.09,98.92,96.88,82.19,80.90,78.55,70.34,69.42,68.23,66.16,61.17,44.70,40.92,40.35,39.71,39.36,36.06,32.04,29.90,29.83,29.79,29.68,29.65,29.61,29.47,29.45,29.37,28.83,27.34,25.67,25.44,25.22,22.82,22.24,21.25,18.03,15.17,14.85,14.25,13.89.

HR-MS(ESI)(M+H)⁺m/z 932.4956, calculation C₅₁H₆₉FN₄O₁₁931.4012

EXAMPLE 10 preparation of Compound S10

Synthetic route to compound S10:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 1.04g of compound j4 was added to the reaction solution, the mixture was heated at 90 ℃ for 8 hours, and the reaction was stopped by TLC (dichloromethane: methanol ═ 10:1) after completion of the reaction. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S10(0.694g, 41.6%).

¹H NMR(500MHz,CDCl₃)δ9.02(dd,J＝19.3,3.9Hz,1H),8.21(t,J＝9.8Hz,1H),7.85(dd,J＝13.9,8.5Hz,1H),7.77(s,1H),7.69(dd,J＝15.3,5.9Hz,1H),7.60(dd,J＝14.4,7.1Hz,1H),7.49(dd,J＝46.9,3.8Hz,1H),7.19(d,J＝8.0Hz,3H),7.12(d,J＝6.6Hz,2H),5.96(dd,J＝15.1,10.3Hz,1H),4.74(dd,J＝22.9,9.9Hz,1H),4.44～4.34(m,1H),4.31(t,J＝7.9Hz,1H),4.03(dd,J＝19.9,10.5Hz,1H),3.84～3.70(m,1H),3.62(dd,J＝14.1,7.2Hz,1H),3.52(s,3H),3.35(d,J＝4.2Hz,1H),3.24(dd,J＝16.2,7.2Hz,1H),3.08～3.02(m,1H),2.60(d,J＝10.5Hz,2H),2.47(s,2H),2.37(s,6H),2.31(s,2H),2.01(s,1H),1.96～1.81(m,3H),1.75(d,J＝21.4Hz,3H),1.44(s,4H),1.33(s,2H),1.26(d,J＝5.9Hz,12H),1.15(d,J＝3.7Hz,3H),0.96(s,3H),0.76(dt,J＝56.5,7.0Hz,3H)

¹³C NMR(100MHz,CDCl₃)δ216.45,202.80,202.32,165.51,157.16,150.43,147.12,143.25,135.44,134.77,130.23,129.80,129.46,128.71,128.32,128.09,127.61,118.24,103.25,98.90,82.16,80.56,78.52,69.91,68.57,66.69,61.52,49.27,47.75,44.66,42.07,40.72,40.41,39.62,39.35,30.34,26.39,25.41,25.19,22.22,21.05,19.86,17.92,14.80,13.89,10.18.

HR-MS(ESI)(M+H)⁺m/z 958.5162, calculation C₅₃H₇₁FN₄O₁₁957.5147

EXAMPLE 11 preparation of Compound S11

Synthetic route to compound S11:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.77g of compound k2 was added to the reaction solution, and the mixture was heated at 90 ℃ for reaction for 8 hours, and the reaction was stopped by TLC (dichloromethane: methanol ═ 10:1) after completion of the reaction. The reaction mixture was cooled to room temperature, spun dry, the residue was dissolved in 35ml of dichloromethane, washed 2 times with 15ml of 5% potassium dihydrogen phosphate and saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain light red solid S11(0.638g, 40.3%).

¹H NMR(500MHz,CDCl₃)δ9.07(d,J＝8.0Hz,1H),8.55(d,J＝6.8Hz,1H),7.93(t,J＝7.3Hz,1H),7.80(d,J＝8.5Hz,1H),7.75(t,J＝7.2Hz,1H),6.77(d,J＝6.9Hz,1H),4.89(d,J＝9.6Hz,1H),4.58(s,2H),4.39(d,J＝6.6Hz,1H),4.06(d,J＝10.2Hz,1H),3.93～3.84(m,1H),3.73～3.64(m,3H),3.58(s,3H),3.51(s,1H),3.43(s,1H),3.15(d,J＝6.5Hz,1H),2.89(s,2H),2.58(s,6H),2.53(s,4H),2.28–2.10(m,3H),1.97(d,J＝14.3Hz,6H),1.80(d,J＝21.3Hz,3H),1.69(dd,J＝14.7,7.6Hz,2H),1.55(s,3H),1.52(s,1H),1.51～1.46(m,2H),1.33(d,J＝6.5Hz,3H),1.29(d,J＝9.0Hz,9H),1.24(d,J＝6.0Hz,3H),1.04(dd,J＝14.9,7.0Hz,8H),0.92(t,J＝6.5Hz,3H).

¹³C NMR(125MHz,CDCl₃)δ216.97,202.67,165.51,157.94,155.98,145.89,137.84,134.43,127.30,126.09,119.06,116.95,103.94,98.60,97.01,82.80,80.53,78.81,70.09,69.06,68.58,66.02,62.02,54.92,49.45,44.79,41.96,40.68,39.27,31.29,26.23,25.47,25.22,24.68,22.38,21.14,20.00,17.90,15.29,14.74,13.89,13.73,10.66.

HR-MS(ESI)(M+H)⁺m/z 871.5188, calculation C₄₇H₇₁FN₄O₁₂870.5154.

EXAMPLE 12 preparation of Compound S12

Synthetic route to compound S12:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.65g of compound l4 was added to the reaction solution, and the mixture was heated at 90 ℃ for reaction for 8 hours, and the completion of the reaction was monitored by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. The reaction mixture was cooled to room temperature, spun dry, the residue was dissolved in 35ml of dichloromethane, washed 2 times with 15ml of 5% potassium dihydrogen phosphate and saturated brine, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, mixing the components, and spin-drying to obtain light red solid S12(0.619g, 40.5%).

¹H NMR(400MHz,CDCl₃)δ9.28(dd,J＝13.3,4.5Hz,1H),8.33(t,J＝8.2Hz,1H),7.86(t,J＝7.5Hz,1H),7.80(t,J＝7.7Hz,1H),7.62(dd,J＝14.5,7.2Hz,1H),7.45(dd,J＝23.1,4.5Hz,1H),4.82(d,J＝10.0Hz,1H),4.37(d,J＝6.2Hz,1H),4.02(t,J＝11.8Hz,1H),3.76(dd,J＝20.2,11.7Hz,1H),3.62～3.59(m,3H),3.38(d,J＝5.6Hz,1H),3.18(d,J＝4.5Hz,3H),3.12(dd,J＝6.8,4.3Hz,1H),2.89(d,J＝7.2Hz,1H),2.64～2.58(m,6H),2.50(s,4H),2.32(s,1H),1.99～1.88(m,3H),1.76(d,J＝4.9Hz,4H),1.52(s,3H),1.36(s,2H),1.35～1.29(m,10H),1.22(d,J＝6.7Hz,2H),1.04～0.99(m,3H),0.85(t,J＝7.4Hz,2H),0.78(t,J＝7.4Hz,2H).

¹³C NMR(100MHz,CDCl₃)δ216.76,201.82,167.38,165.37,165.14,150.93,136.96,132.92,131.30,129.56,129.47,128.97,128.81,128.18,127.93,127.35,103.32,82.12,80.61,78.66,69.92,68.55,66.46,61.42,49.21,48.10,44.65,41.72,40.76,39.79,37.75,30.43,30.29,29.13,26.25,25.16,23.74,22.22,21.05,19.89,18.52,15.46,14.27,13.96,10.46.

HR-MS(ESI)(M+H)⁺m/z 829.4718, calculation C₄₄H₆₅FN₄O₁₀828.4685

EXAMPLE 13 preparation of Compound S13

Synthetic route to compound S13:

0.724g, 1mmol of compound 8 was dissolved in 9ml of acetonitrile and 1ml of water, 3.0mmol, 0.87g of compound m4 was added to the reaction solution, and the mixture was heated at 90 ℃ for reaction for 8 hours, and the completion of the reaction was monitored by TLC (dichloromethane: methanol ═ 10:1), i.e., the reaction was stopped. And cooling the reaction liquid to room temperature, and spin-drying to obtain a light yellow solid. The residue was dissolved in 35ml of dichloromethane, washed with 15ml of 5% potassium dihydrogen phosphate and saturated brine for 2 times, separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. Column chromatography, dichloromethane infiltration, dichloromethane: eluting with 50: 1-10: 1 methanol, combining the components, and spin-drying to obtain white foamy solid S13(0.652g, 40.7%).

¹H NMR(500MHz,CDCl₃)δ9.55(d,J＝8.3Hz,1H),8.09(d,J＝7.3Hz,1H),7.88(t,J＝7.7Hz,1H),7.79～7.69(m,2H),7.45(d,J＝7.2Hz,2H),7.29(t,J＝7.2Hz,2H),7.24(d,J＝7.3Hz,1H),6.40(d,J＝7.4Hz,1H),4.87(s,2H),4.80(d,J＝9.6Hz,2H),4.45(dt,J＝13.5,6.3Hz,1H),4.39～4.29(m,2H),4.02(d,J＝10.5Hz,1H),3.89～3.78(m,1H),3.54(dt,J＝17.6,9.4Hz,4H),3.39(s,1H),3.27(t,J＝8.3Hz,1H),3.18～3.06(m,1H),2.60(d,J＝45.7Hz,3H),2.40(s,5H),2.37(s,4H),2.20(s,4H),1.96(dt,J＝14.1,8.6Hz,2H),1.89(d,J＝14.9Hz,2H),1.76(d,J＝21.4Hz,3H),1.71～1.59(m,2H),1.50(s,3H),1.30(d,J＝6.6Hz,6H),1.25(d,J＝5.8Hz,12H),1.17(d,J＝6.5Hz,3H),0.98(d,J＝6.6Hz,3H),0.87(d,J＝7.0Hz,3H).

¹³C NMR(125MHz,CDCl₃)δ217.06,202.78,166.61,157.57,145.10,137.33,135.96,134.24,128.83,127.66,127.35,127.08,119.01,116.48,103.96,98.88,96.83,82.63,80.46,78.72,70.09,69.24,65.86,61.49,52.31,48.97,46.92,44.52,41.24,40.81,40.37,39.47,38.99,38.68,29.73,29.35,27.45,25.28,25.10,22.93,22.63,22.12,21.10,19.66,17.74,15.07,14.55,14.05,13.66,10.93,10.45.

HR-MS(ESI)(M+H)⁺m/z 919.5201,calcd for C₅₁H₇₁FN₄O₁₀918.5190.

Experimental examples section

Experimental example 1 pharmacological experiment: in vitro antibacterial Activity test

The invention provides the application of the compound in the aspect of antibiosis by utilizing the in-vitro antibacterial activity experiment.

(1) The experimental method comprises the following steps: culture medium and incubation conditions

Incubating staphylococcus in CAMHB culture medium at 35 deg.C for 16-20 hr; streptococcus is incubated in CAMHB medium with 5% horse serum at 35 deg.C for 20-24 h.

Minimum Inhibitory Concentration (MIC) determination

Standard two-fold broth dilution was used. The concentration range of the antibacterial drug is 64-0.004 mg/L. The final concentration of the tested bacterial liquid is about 1 multiplied by 10⁵CFU/ml。

(2) Control drug: clarithromycin (Cla), telithromycin (Teli)

Clinical isolate 11 strains:

methicillin-susceptible staphylococcus epidermidis-ATCC 12228, 16-4

Methicillin-resistant staphylococcus epidermidis-16-5

Methicillin-sensitive Staphylococcus aureus (ATCC 29213, 15, 16-1)

Methicillin-resistant staphylococcus aureus-ATCC 33591, 16-30

Vancomycin-sensitive enterococcus faecalis-ATCC 33591, 16-6

Vancomycin resistant enterococcus faecalis-ZTCC 51299, ATCC51575

Vancomycin-sensitive enterococcus faecium-16-5

Vancomycin resistant enterococcus faecium-ATCC 700221, 12-1

Erythromycin sensitive Streptococcus pyogenes-ATCC 19615

Erythromycin drug-resistant streptococcus pyogenes-12-1

Erythromycin sensitive Streptococcus pneumoniae-ATCC 49616

Erythromycin drug-resistant streptococcus pneumoniae-12-3

Escherichia coli ATCC25922, ATCC35218, 16-1 and 16-7

Klebsiella pneumoniae-ATCC 700603, 7, ATCCBA-2146, 16-2, 16-4

Pseudomonas aeruginosa-ATCC 29853, PA01, 16-11

Acinetobacter baumannii-ATCC 19606

Enterobacter cloacae-ATCC 43560

Serratia marcescens-ATCC 21074

Citrobacter freudenreichii-ATCC 43864

Klebsiella pneumoniae-ATCC 31052

Proteus mirabilis-ATCC 49565

Pseudomonas maltophilia-ATCC 13636

Shigella flexneri-ATCC 12022

TABLE 1 MIC results (. mu.g/L) for compounds S1-S13 and control against 13 strains of bacteria

Experiments prove that the compound sample with the structural formula of S1-S13, prepared by the method, has broad spectrum and simultaneously inhibits outstanding antibacterial activity and drug-resistant activity of gram-positive bacteria and gram-negative bacteria.

The foregoing description is of the preferred embodiments of the present invention only, and thus all features and methods that are described in the claims of the present invention are included in the claims of the present invention.

Claims

1. Erythromycin A antibiotic derivatives shown as general formula I and pharmaceutically acceptable salts thereof,

y is selected from substituted or unsubstituted quinoline, wherein the substituents are selected from hydrogen, amino, nitro, C1-3 alkoxy.

2. Erythromycin A-type antibiotic derivatives and pharmaceutically acceptable salts thereof according to claim 1,

x is selected from hydrogen, C1-6 alkyl, C6-10 aryl C1-4 alkyl, C1-6 alkanoyl, C6-10 aryl C1-4 alkanoyl and cinnamoyl.

3. Erythromycin A-type antibiotic derivatives and pharmaceutically acceptable salts thereof according to claim 1,

wherein X is selected from hydrogen, C1-6 alkyl, C6-10 aryl C1-4 alkyl, C1-6 alkanoyl, C6-10 aryl C1-4 alkanoyl and cinnamoyl.

4. An erythromycin A class antibiotic derivative and its pharmaceutically acceptable salts according to any one of claims 1-3, selected from the following:

compound S1

Compound S2

Compound S3

Compound S4

Compound S5

Compound S6

Compound S7

Compound S8

Compound S9

Compound S10

Compound S11

Compound S12

Compound S13

5. A process for the preparation of an erythromycin A derivative according to any one of claims 1 to 4, characterized by comprising the following steps:

starting from clarithromycin, the reaction is carried out in 7 steps to obtain a macrolide intermediate compound 8 which is properly protected; then, the macrolide intermediate compound 8 and a primary amine side chain are subjected to addition reaction to obtain a corresponding erythromycin A antibiotic derivative;

wherein the substituent X, Y is as defined in any one of claims 1 to 4.

6. A pharmaceutical composition comprising at least one erythromycin A class antibiotic derivative according to any one of claims 1-4 and a pharmaceutically acceptable carrier.

7. Use of erythromycin A antibiotic derivatives and pharmaceutically acceptable salts thereof as claimed in any one of claims 1 to 4 for the preparation of a medicament for inhibiting bacteria.

8. Use according to claim 7, characterized in that said bacteria are selected from the group consisting of gram-positive bacteria, gram-negative bacteria.

9. Use according to claim 8,