CN102786570A

CN102786570A - Macrolide compounds, preparation method thereof, application thereof and intermediate thereof

Info

Publication number: CN102786570A
Application number: CN2011101293400A
Authority: CN
Inventors: 沈舜义; 袁芳; 葛涵; 张志宏; 刘毓彬
Original assignee: Shanghai Institute of Pharmaceutical Industry
Current assignee: Shanghai Institute of Pharmaceutical Industry
Priority date: 2011-05-18
Filing date: 2011-05-18
Publication date: 2012-11-21
Anticipated expiration: 2031-05-18
Also published as: CN102786570B

Abstract

The invention discloses macrolide compounds with the general structure represented by formula I, a preparation method thereof, an application thereof and an intermediate thereof. The macrolide compounds have very strong activities on most test G<+> bacteria, wherein the activities of two types of compounds to partial gram-positive bacteria are higher than the activities of Azithromycin to the partial gram-positive bacteria.

Description

Macrolide compound, preparation method, application and intermediate thereof

Technical Field

The invention particularly relates to a macrolide compound, and a preparation method, application and an intermediate thereof.

Background

Macrolide antibiotics have been applied to infection resistance of various clinical departments for over 50 years, have the advantages of high safety, low price, good oral absorption and the like, and play an important role in treating infectious diseases.

The chemical structure modification work of macrolide antibiotics is based on the research of the action mechanism of medicaments and goes through three development stages. In the early stage, the compound is mainly modified into esters and salts to improve bitter taste, enhance stability and improve bioavailability. In the 70 s of the 20 th century, the acid inactivation mechanism of erythromycin A was clarified, and the modification of erythromycin A after that mainly aims at active sites participating in acid degradation to obtain a second generation macrolide compound represented by clarithromycin and azithromycin, which has improved acid stability and improved pharmacokinetic properties. With the increasing problem of drug-resistant bacteria worldwide, the second generation macrolide antibiotics show the weakness of poor activity to the drug-resistant bacteria. In the 90 s, researches show that cladinose is a main group for inducing bacteria to generate drug resistance, and the position is modified to obtain third-generation macrolide antibiotics such as ketolide, 4' -carbamate, acyl lactone, 2, 3-anhydro-ene lactone and the like, so that the induced drug resistance of the bacteria is overcome, and the activity of the antibiotics on drug-resistant bacteria is strong.

The development of macrolide antibiotics is expected, and the search for effective macrolide antibiotics against drug-resistant bacteria is still the focus and focus of research.

Disclosure of Invention

The invention aims to solve the technical problem of providing macrolide compounds, a preparation method, application and an intermediate thereof, which are completely different from the prior art. Macrolide Compounds of the invention for most of the tested G⁺The bacteria have strong activity, wherein the activity of the two compounds on partial gram-positive bacteria is higher than that of azithromycin.

Therefore, the invention relates to macrolide compounds shown in a formula I;

wherein R is C₁～C₃Alkyl, or H; r^aIs composed of

In the 3-positionRepresents a single bond or a double bond, in which case R represents a double bond^bIn the absence of, which is a single bond, R^bIs composed of

R^cIs substituted or unsubstituted C₁～C₃Alkyl (preferably methyl, ethyl or isopropyl), substituted or unsubstituted C₆～C₁₀Aryl, or substituted or unsubstituted C₂～C₄Alkenyl (preferably vinyl); wherein,substituted C₆～C₁₀The substituent in the aryl is nitro or halogen (halogen is preferably C1), and the substituent in the aryl can be ortho, para or meta; substituted C₁～C₃The substituent in the alkyl group being C₆～C₁₀Aryl, or C₄～C₈Heteroaryl (preferably 2-thienyl or 3-indolyl), wherein the heteroatom is N, O or S; substituted C₂～C₄The substituent in the alkenyl group is C₆～C₁₀An aryl group;

X¹and X²Independently of one another, halogen, such as fluorine, chlorine, bromine or iodine, X¹And X²The positions of (a) can be 2 'and 5'; preferably, X¹And X²The same;

R^dand R^eIndependently is C₁～C₃Alkyl (preferably ethyl);

n is 1 to 3 (preferably 2).

In the present invention, the compound I is preferably of any one of the following structures:

wherein Ra and Rc are as defined above.

In the present invention, the compound I is more preferably any one of the following compounds:

in Ia, R^cIs methyl, phenyl, p-nitrophenyl, p-chlorophenyl, benzyl, phenethyl, styryl, thiophene-2-methyl, indole-3-methyl or indole-3-propyl;

in Ib, R^cIs phenyl, p-nitrophenyl, p-chlorophenyl, benzyl, phenethyl, styryl or isopropyl;

in Ic, R^aIs composed of

For the preparation of the compound I of the present invention, those skilled in the art can refer to the prior art, and the corresponding target product can be obtained.

The invention further relates to a preparation method of the compound I, which is any one of the following methods:

(I) when the object compound I, R^aIs composed ofR is C₁～C₃When in alkyl, the compound II is subjected to a reaction of removing the acyl protecting group of the hydroxyl;

R^c1is R^cOr a substituent on an acyl protecting group as is conventional in the art;

(II) when the object compound I, R^aIs composed of

R is H or C₁～C₃When alkyl, compounds II' and R^aX is subjected to nucleophilic substitution reaction as shown in the specification;

wherein X is a leaving group conventional in such nucleophilic substitution reactions in the art, such as a halogen (preferably chlorine, bromine or iodine).

In both methods, the methods and conditions of the reactions involved can be conventional in the corresponding classes of reactions in the art; the definitions of the groups are as described above unless otherwise indicated.

The compounds I of the present invention can be prepared according to the prior art using any suitable organic synthesis method. The preferred preparation of compounds I according to the invention is illustrated below:

the preparation of compound Ia series, the reaction scheme is as follows:

the method comprises the following steps of taking clarithromycin as a starting material, reacting with hydrazine acetate to generate 9-hydrazone clarithromycin, carrying out double acylation, and removing 2' -acylation protection to obtain a target compound.

Wherein each group is as defined above. The compounds Ia can be prepared by the skilled person according to the prior art and the general knowledge, with reference to the above reaction schemes.

The preparation of compound Ib series, the reaction route is as follows:

the method comprises the following steps of taking clarithromycin as a starting material, reacting with hydrazine acetate to generate 9-hydrazone clarithromycin, carrying out double acylation, removing cladinose, oxidizing, and carrying out deacylation to obtain a target compound.

Wherein each group is as defined above. The compounds Ib can be prepared by the skilled worker according to the prior art and the general knowledge by referring to the above-mentioned reaction schemes.

(III) preparation of compound Ic series, the synthetic route is as follows:

taking erythromycin thiocyanate A as a raw material, and obtaining a target compound through three steps of reaction of hydrazone formation, thiocyanate radical removal and alkylation.

Wherein each group is as defined above. The compound Ic can be prepared by those skilled in the art according to the prior art and conventional knowledge by referring to the above reaction scheme.

The present invention therefore further relates to the intermediate compounds IIa, IIb or IIIb for the preparation of the above-mentioned compounds I:

wherein each group is as defined above.

The invention also relates to the application of the compound I in preparing antibiotic medicaments.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: macrolide Compounds of the invention for most of the tested G⁺The bacteria have strong activity, wherein the activity of the two compounds on partial gram-positive bacteria is higher than that of azithromycin.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

The specific compounds involved in the examples of the invention are as follows:

TABLE 1 Ia series of compounds

TABLE 2 Ib series of compounds

Compound 18

Compound 19

Preparation examples:

in the following examples:

thin Layer Chromatography (TLC): silica gel HSGF254 (yellow silica gel development laboratory, Taiwan, sesame 32600

Color development: ultraviolet light (254nm, 365nm) and color developing agent (100 ml prepared from 1g cerium sulfate, 2.5g sodium molybdate and 10% sulfuric acid solution)

Column chromatography silica gel: thin layer chromatography silica gel H (Qingdao ocean chemical products of Co., Ltd.)

NMR: Varian-Inova-400 nuclear magnetic resonance apparatus

MS: MAT212 magnetic mass spectrometer

IR: NEXUS-670 model infrared spectrometer, potassium bromide tabletting method

HPLC: agilentl100GC and ultraviolet absorption detector

Description of English abbreviations

EXAMPLE 1 hydrazine acetate (intermediate 1)

To a 100ml round bottom flask was added 85% hydrazine hydrate (28.5ml, 0.5mol), stirred in an ice bath and glacial acetic acid (28.6ml, 0.5mol) was slowly added dropwise, controlling the temperature at 0-10 ℃. After the dropwise addition, the reaction was stirred at room temperature for 40 min. After water was removed by distillation under reduced pressure, chloroform/ethanol solution (1: 1, 20ml) was added to the remaining liquid, and the mixture was crystallized by stirring in an ice bath, then further crystallized in a refrigerator at 4 ℃ and evaporated to dryness under reduced pressure to obtain a white solid (41.6g, 90.5%).

Example 29-Hydrazone Clarithromycin (intermediate 2)

Hydrazine acetate (37g, 0.4mol) and clarithromycin (10g, 13.36mmol) were placed in a round-bottomed flask, and methanol (80ml) was added thereto, followed by heating and refluxing for 46 hours. Vacuum distilling, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, separating organic phase, extracting water phase with dichloromethane three times, combining organic phases, washing with water, washing with saturated salt solution, drying with anhydrous sodium carbonate, filtering, and evaporating filtrate under reduced pressure to obtain white solid (9.68g, 95.1%).

Example 32' -O-acetyl-9-acetylhydrazone clarithromycin (intermediate 3)

Dissolving 9-hydrazone clarithromycin (3.00g, 3.94mmol) in dichloromethane (15mL), adding acetic anhydride (1.12mL, 11.85mmol), reacting at normal temperature for 2h, adding water, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to obtain a white bubble (3.28g, 98.5%). (Small amount of experiments have used acetic anhydride and acetic acid + DCC and 9-hydrazone clarithromycin reaction process and reaction products the same, so the amplification reaction using acetic anhydride and 9-hydrazone clarithromycin, convenient)

MS(ESI⁺，m/e)：846.52[M+H]⁺

Example 49-Acetylhydrazone clarithromycin (Compound 1)

Dissolving 2' -O-acetyl-9-acetyl hydrazone clarithromycin (intermediate 3) (1.00g, 1.18mmol) in methanol (5mL), heating and refluxing for 6h, evaporating to dryness under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating to dryness under reduced pressure, and performing column chromatography to separate [ ethyl acetate, petroleum ether and diethylamine (5: 12: 1) ], thus obtaining a light yellow solid (0.41g, 43.2%).

MS(ESI⁺，m/e)：804.51[M+H]⁺

¹HNMR(400MHz，CDCl₃)δ(ppm)：8.58(s，1H，9＝N-NH-)，2.19(s，3H，9-CO-CH₃)

¹³CNMR(400MHz，CDCl₃)δ(ppm)：172.56(9-NH-CO-)，166.92(9＞C＝N-)

Example 52' -O-benzoyl-9-benzoylhydrazone clarithromycin (intermediate 4)

Benzoic acid (4.00g, 32.75mmol) and DCC (7.00g, 33.93mmol) were dissolved in dichloromethane (50mL) and reacted at room temperature for 30min, followed by addition of 9-hydrazone clarithromycin (5.00g, 6.56mmol) and reaction at room temperature for 60 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out organic phase, extracting water phase with dichloromethane three times, combining organic phases, washing with water, washing with saturated salt solution, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain white crude product (9.35g, containing part dicyclohexylurea, the same below).

MS(ESI⁺，m/e)：970.56[M+H]⁺

Example 69-Benzoylhydrazone clarithromycin (Compound 2)

Dissolving the crude 2' -O-benzoyl-9-benzoyl hydrazone clarithromycin (intermediate 4, 1.50g and 1.05mmol) in methanol (6mL), heating and refluxing for 2.5h, evaporating to dryness under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography separation [ ethyl acetate, petroleum ether and diethylamine (5: 7: 0.6) ], so as to obtain white foams (0.40g, from the intermediate 2 to the intermediate 4, and finally obtaining the total yield of the compound 2, which is 44.0%).

MS(ESI⁺，m/e)：866.47[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.69(s, 1H, 9 ═ N-NH-), 7.40-7.90(m, 5H, 9-position benzene ring)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 166.41(9-NH-CO-), 163.30(9 > C ═ N-), 127.21-134.38 (6 carbons on the 9-position phenyl ring)

Example 72' -O-benzoyl-3-O-descladinose-3-hydroxy-9-benzoylhydrazone clarithromycin (intermediate 5)

Dissolving the crude 2' -O-benzoyl-9-benzoyl hydrazone clarithromycin (intermediate 4, 2.00g and 1.40mmol) in 1% ethanol hydrochloride solution (20mL), reacting at normal temperature for 12h, adding water, dichloromethane and 3N NaOH to adjust the pH value to 9.7, separating out an organic phase, extracting the aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (1.66 g).

MS(ESI⁺，m/e)：812.46[M+H]⁺

Example 82' -O-benzoyl-3-O-descladinose-3-oxo-9-benzoylhydrazone clarithromycin (intermediate 6)

The crude 2' -O-benzoyl-3-O-decladinose-3-hydroxy-9-benzoylhydrazone clarithromycin (intermediate 5, 1.66g, 1.40mmol) and EDC. HCl (2.63g, 13.71mmol) were dissolved in anhydrous dichloromethane (17mL), DMSO (2.62mL, 36.84mmol) was added, TFA. Py (1.31g, 6.75 mm) was slowly added dropwiseol) in dichloromethane, N₂Reacting at normal temperature for 1h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (1.66 g).

MS(ESI⁺，m/e)：810.45[M+H]⁺

Example 93-O-decladinose-3-oxo-9-benzoylhydrazone clarithromycin (Compound 11)

Dissolving the crude 2' -O-benzoyl-3-O-decladinose-3-oxo-9-benzoyl hydrazone clarithromycin (intermediate 6, 1.66g and 1.40mmol) in methanol (9mL), heating and refluxing for 2h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography separation [ ethyl acetate, petroleum ether and diethylamine (5: 8: 0.6) ], thus obtaining a pale yellow foam (0.56g, from the intermediate 2 to the intermediates 4, 5 and 6, and finally obtaining the total yield of the compound 11 of 56.6%).

MS(ESI⁺，m/e)：706.57[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.63(s, 1H, 9 ═ N-NH), 7.41-7.86(m, 5H, 9-position phenyl ring)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 202.32.72(3 > C ═ O), 165.34(9-NH-CO-), 163.32(9 > C ═ N-), 104.39-134.23 (6 carbons on the benzene ring of the side chain at position 9)

Example 102' -O-p-nitrobenzoyl-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 7)

P-nitrobenzoic acid (3.29g, 19.69mmol) and DCC (4.22g, 20.45mmol) were dissolved in dichloromethane (30mL) and reacted at room temperature for 30min, followed by addition of 9-hydrazone clarithromycin (3.00g, 3.94mmol) and reaction at room temperature for 1 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow crude product (4.54 g).

MS(ESI⁺，m/e)：1060.53[M+H]⁺

Example 119-p-nitrobenzoyl hydrazone clarithromycin (Compound 3)

Dissolving the crude 2' -O-p-nitrobenzoyl-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 7, 1.30g and 1.13mmol) in methanol (6.50mL), heating and refluxing for 1h, evaporating to dryness under reduced pressure, adding water and dichloromethane and 3N NaOH to adjust the pH value to 9.7, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate: petroleum ether: diethylamine (5: 4: 0.4) ] to obtain yellow bubbles (0.27g, wherein the total yield of the intermediate 2, the intermediate 7 and the compound 3 is 26.5%).

MS(ESI⁺，m/e)：911.20[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.24(s, 1H, 9 ═ N-NH), 8.02-8.29(m, 4H, 9-position phenyl ring)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 149.24(9-NH-CO-), 128.64(9 > C ═ N-), 123.20-123.70 (6 carbons on the benzene ring of the 9-position side chain)

Example 122' -O-p-nitrobenzoyl-3-O-decoladinose-3-hydroxy-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 8)

Dissolving the crude 2' -O-p-nitrobenzoyl-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 7, 2.00g and 1.73mmol) in 1% hydrochloric acid ethanol solution (20mL), reacting at normal temperature for 7h, adding water, dichloromethane and 3N NaOH to adjust the pH value to 9.7, separating out an organic phase, extracting the aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow solid (1.70 g).

MS(ESI⁺，m/e)：902.43[M+H]⁺

EXAMPLE 132' -O-p-nitrobenzoyl-3-O-descladinose-3-oxo-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 9)

The crude 2' -O-p-nitrobenzoyl-3-O-decladinose-3-hydroxy-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 8, 1.70g, 1.73mmol) and EDC. HCl (2.42g, 12.64mmol) were dissolved in anhydrous dichloromethane (17mL), DMSO (2.42mL, 34.02mmol) was added, and a solution of TFA. Py (1.20g, 6.24mmol) in dichloromethane, N, was slowly added dropwise₂Reacting at normal temperature for 1h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow solid (1.69 g).

MS(ESI⁺，m/e)：900.42[M+H]⁺

Example 143-O-descladinose-3-oxo-9-p-nitrobenzoyl hydrazone clarithromycin (Compound 12)

Dissolving the crude 2' -O-p-nitrobenzoyl-3-O-decoladinose-3-oxo-9-p-nitrobenzoyl hydrazone clarithromycin (intermediate 9, 1.69g and 1.73mmol) in methanol (9mL), heating and refluxing for 2h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and separating by column chromatography [ methanol: chloroform (1: 3) ], thus obtaining a brown yellow foam (0.32g, from the intermediate 2 to the intermediates 7, 8 and 9, and finally obtaining the total yield of the compound 12 of 24.6%).

MS(ESI⁺，m/e)：751.31[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.90(s, 1H, 9 ═ N-NH), 7.98-8.25(m, 4H, 9-position benzene ring)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 202.2(3 > C ═ O), 168.0(9-NH-CO-), 160.3(9 > C ═ N-), 123.39-149.56 (6 carbons on the benzene ring of the side chain at position 9)

Example 152' -O-p-chlorobenzoyl-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 10)

P-chlorobenzoic acid (3.08g, 19.67mmol) and DCC (4.22g, 20.45mmol) were dissolved in dichloromethane (30mL) and reacted at room temperature for 30min, and 9-hydrazone clarithromycin (3.00g, 3.94mmol) was added and reacted at room temperature for 24 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white crude product (6.02 g).

MS(ESI⁺，m/e)：1038.48[M+H]⁺

Example 169-p-chlorobenzoylhydrazone clarithromycin (Compound 4)

Dissolving the crude 2' -O-p-chlorobenzoyl-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 10, 2.00g and 1.31mmol) in methanol (10mL), heating and refluxing for 5h, evaporating to dryness under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate, petroleum ether and diethylamine (5: 7.5: 0.6) ], thus obtaining a light yellow solid (0.45g, wherein the total yield of the intermediate 2, the intermediate 10 and the compound 4 is 38.4%).

MS(ESI⁺，m/e)：900.40[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.74(s, 1H, 9 ═ N-NH), 7.80(d, J ═ 8Hz, 2H, 9-position phenyl ring 3, 5-H), 7.39(d, J ═ 8.4Hz, 2H, 9-position phenyl ring 2, 6-H)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 167.06(9-NH-CO-), 162.25(9 > C ═ N-), 128.63-156.93 (6 carbons on the benzene ring of the 9-position side chain)

EXAMPLE 172' -O-p-chlorobenzoyl-3-O-decoladinose-3-hydroxy-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 11)

Dissolving the crude 2' -O-p-chlorobenzoyl-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 10, 2.00g and 1.31mmol) in 1% hydrochloric acid ethanol solution (20mL), reacting at normal temperature for 17h, adding water, dichloromethane and 3N NaOH to adjust the pH value to 9.7, separating out an organic phase, extracting the aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (1.68 g).

MS(ESI⁺，m/e)：880.38[M+H]⁺

EXAMPLE 182' -O-p-chlorobenzoyl-3-O-decladinose-3-oxo-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 12)

The crude 2' -O-p-chlorobenzoyl-3-O-decladinose-3-hydroxy-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 11, 1.68g, 1.31mmol) and EDC. HCl (2.45g, 12.79mmol) were dissolved in anhydrous dichloromethane (17mL), DMSO (2.44mL, 34.38mmol) was added, and a solution of TFA. Py (1.22g, 6.30mmol) in dichloromethane, N, was slowly added dropwise₂Reacting at normal temperature for 1h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (1.66 g).

MS(ESI⁺，m/e)：878.37[M+H]⁺

Example 193-O-decladinose-3-oxo-9-p-chlorobenzoylhydrazone clarithromycin (Compound 13)

Dissolving the crude 2' -O-p-chlorobenzoyl-3-O-decladinose-3-oxo-9-p-chlorobenzoyl hydrazone clarithromycin (intermediate 12, 1.66g and 1.31mmol) in methanol (9mL), heating and refluxing for 2h, evaporating under reduced pressure, adding water and dichloromethane, 3N NaOH to adjust the pH value to 9.7, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography separation [ methanol: chloroform (1: 16) ], thus obtaining a yellow solid (0.19g, from the intermediate 2 to the intermediates 10, 11 and 12, and finally obtaining the total yield of the compound 13 of 19%).

MS(ESI⁺，m/e)：740.33[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.7(s, 1H, 9 ═ N-NH), 7.37-7.78(m, 4H, 9-position benzene ring)

¹³CNMR(400MHz，CDCl₃)δ(ppm)：202.51(3＝O)，172.52(9-NH-CO-)，170.14(9＞C＝N-)，103.92-137.55(96 carbons in the side-chain benzene ring)

Example 202' -O-phenylacetyl-9-phenylacetylhydrazone clarithromycin (intermediate 13)

Phenylacetic acid (5.36g, 39.37mmol) and DCC (8.93g, 43.28mmol) were dissolved in dichloromethane (60mL) and reacted at room temperature for 30min, followed by addition of 9-hydrazone clarithromycin (6.00g, 7.87mmol) and reaction at room temperature for 1 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a light yellow crude product (9.96 g).

MS(ESI⁺，m/e)：998.59[M+H]⁺

Example 219-Phenylacetylhydrazone clarithromycin (Compound 5)

Dissolving the crude 2' -O-phenylacetyl-9-phenylacetylhydrazone clarithromycin (intermediate 13, 4.50g and 3.56mmol) in methanol (23mL), heating and refluxing for 2.5h, evaporating to dryness under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate to dryness under reduced pressure to obtain 4.45g, performing column chromatography (ethyl acetate, petroleum ether, diethylamine (5: 8: 0.6)) to obtain a light yellow substance (0.45g, from intermediate 2 to intermediate 13, and finally obtaining the total yield of the compound 5 of 50.0%).

MS(ESI⁺，m/e)：880.51[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.58(s, 1H, 9 ═ N-NH), 7.22 to 7.36(m, 5H, 9-phenyl ring), 4.54(s, 2H, 9-CO-CH)₂-)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 172.56(9-NH-CO-), 166.92(9 > C ═ N-), 126.74-129.30 (6 carbons on the benzene ring of the 9 th side chain), 32.75 (carbons between the acyl group of the 9 th side chain and the benzene ring)

EXAMPLE 222' -O-phenylacetyl-3-O-descladinose-3-hydroxy-9-phenylacetylhydrazone clarithromycin (intermediate 14)

Dissolving the crude 2' -O-phenylacetyl-9-phenylacetylhydrazone clarithromycin (intermediate 13, 1.73g and 1.37mmol) in 1% ethanol hydrochloride solution (17mL), reacting at normal temperature for 21h, adding water, dichloromethane and 3N NaOH to adjust the pH value to 9.7, separating out an organic phase, extracting the aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to obtain a white foam (1.44 g).

MS(ESI⁺，m/e)：840.49[M+H]⁺

Example 232' -O-Phenylacetyl-3-O-decladinose-3-oxo-9-phenylacetylhydrazone clarithromycin (intermediate 15)

The crude 2' -O-phenylacetyl-3-O-decladinose-3-hydroxy-9-phenylacetylhydrazone clarithromycin (intermediate 14, 1.44g, 1.37mmol) and EDC. HCl (2.20g, 11.50mmol) were dissolved in anhydrous dichloromethane (15mL), DMSO (2.19mL, 30.89mmol) was added, and a solution of TFA. Py (1.09g, 5.66mmol) in dichloromethane, N, was slowly added dropwise₂Reacting at normal temperature for 1h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow solid (1.30 g).

MS(ESI⁺，m/e)：838.48[M+H]⁺

Example 243-O-decladinose-3-oxo-9-phenylacetylhydrazone clarithromycin (Compound 14)

Dissolving the crude 2' -O-phenylacetyl-3-O-decladinose-3-oxo-9-phenylacetylhydrazone clarithromycin (intermediate 15, 1.30g and 1.37mmol) in methanol (8mL), heating and refluxing for 3h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography separation [ ethyl acetate, petroleum ether and diethylamine (5: 8: 0.6) ], thus obtaining a light yellow foam (0.40g, from the intermediate 2 to the intermediates 13, 14 and 15, and finally obtaining the total yield of the compound 14 of 32.0%).

MS(ESI⁺，m/e)：720.41[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 10.66(s, 1H, 9 ═ N-H), 7.21-7.30(m, 5H, 9-phenyl ring), 2.55(s, 2H, 9-CO-CH)₂-Ph)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 205.79(3 ═ O), 172.33(9-NH-CO-), 167.18(9 > C ═ N-), 104.30-134.40 (6 carbons on the benzene ring of the side chain at position 9), 38.29 (carbon between the acyl group of the side chain at position 9 and the benzene ring)

Example 252' -O- β -Phenylbenzoyl-9- β -Phenylacylhydrazone clarithromycin (intermediate 16)

Beta-phenylpropionic acid (5.90g, 39.29mmol) and DCC (8.45g, 40.95mmol) were dissolved in dichloromethane (60mL) and reacted at room temperature for 30min, followed by addition of 9-hydrazone clarithromycin (6.00g, 7.87mmol) and reaction at room temperature for 24 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a light yellow crude product (10.29 g).

MS(ESI⁺，m/e)：1026.62[M+H]⁺

Example 269- β -Phenylhydrazone clarithromycin (Compound 6)

Dissolving the crude 2' -O-beta-phenylpropionyl-9-beta-phenylpropionylhydrazone clarithromycin (intermediate 16, 2.19g and 1.67mmol) in methanol (10mL), heating and refluxing for 5h, evaporating to dryness under reduced pressure, adding water and dichloromethane and 3N NaOH to adjust the pH value to 9.7, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate, petroleum ether and diethylamine (5: 10: 0.6) ], so as to obtain a yellowish foamy substance (0.61g, wherein the total yield of the compound 6 from the intermediate 2 to the intermediate 16 is 40.7%).

MS(ESI⁺，m/e)：894.27[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.60(s, 1H, 9 ═ N-NH), 7.15 to 7.29(m, 5H, 9-phenyl ring), 2.7 to 3.1, 2.2 to 2.5(m, 4H, 9-CO-CH)₂CH₂-Ph)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 174.03(9-NH-CO-), 167.08(9 > C ═ N-), 126.01-140.93 (6 carbons on the benzene ring of the 9-position side chain), 33.36, 30.44 (two alkyl carbons between the 9-position acyl structure and the benzene ring)

Example 272' -O- β -phenylpropionyl-3-O-decoladinose-3-hydroxy-9- β -phenylpropionylhydrazone clarithromycin (intermediate 17)

Dissolving the crude 2' -O-beta-phenylpropionyl-9-beta-phenylpropionylhydrazone clarithromycin (intermediate 16, 2.20g and 1.68mmol) in 1% hydrochloric acid ethanol solution (22mL), reacting at normal temperature for 24 hours, adding water, dichloromethane and 3N NaOH into filtrate to adjust the pH value to 9.7, separating out an organic phase, extracting a water phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow sticky solid (1.83 g).

MS(ESI⁺，m/e)：868.52[M+H]⁺

EXAMPLE 282' -O- β -phenylpropionyl-3-O-decladinose-3-oxo-9- β -phenylpropionylhydrazone clarithromycin (intermediate 18)

The crude 2' -O-. beta. -phenylpropionyl-3-O-decladinose-3-hydroxy-9-. beta. -phenylpropionylhydrazone clarithromycin described above (intermediate 17, 1.83g, 1.68mmol) and EDC. HCl (2.71g, 14.14mmol) were dissolved in anhydrous dichloromethane (18mL), DMSO (2.70mL, 37.99mmol) was added, and a solution of TFA. Py (1.35g, 6.97mmol) in dichloromethane, N, was slowly added dropwise₂Reacting at normal temperature for 0.5h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow sticky solid (1.82 g).

MS(ESI⁺，m/e)：866.51[M+H]⁺

Example 293-O-decoladinose-3-oxo-9-. beta. -hydrocinnamoylhydrazone clarithromycin (Compound 15)

Dissolving the crude 2' -O-phenylpropionyl-3-O-decladinose-3-oxo-9-beta-phenylpropionylhydrazone clarithromycin (intermediate 18, 1.82g and 1.68mmol) in methanol (9mL), heating and refluxing for 3h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and separating by column chromatography [ ethyl acetate: petroleum ether: diethylamine (5: 8: 0.6) ] to obtain a pale yellow foam (0.56g, from intermediate 2, to intermediates 16, 17 and 18, and finally to the total yield of the compound 15 being 45.5%).

MS(ESI⁺，m/e)：734.05[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.80(s, 1H, 9 ═ N-NH), 7.14 to 7.27(m, 5H, 9-phenyl ring), 2.50 to 2.90, 3.0 to 3.3(m, 4H, 9-CO-CH)₂CH₂-Ph)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 205.81(3 > C ═ O), 169.86(9-NH-CO-), 166.64(9 > C ═ N-), 126.05-140.89 (6 carbons on the benzene ring of the side chain at position 9), 34.29, 28.36 (two alkyl carbons between the acyl structure at position 9 and the benzene ring)

Example 302' -O-cinnamoyl-9-cinnamoylhydrazone clarithromycin (intermediate 19)

Cinnamic acid (5.83g, 39.35mmol) and DCC (8.93g, 43.28mmol) were dissolved in dichloromethane (60mL) and reacted at room temperature for 30min, followed by addition of 9-hydrazone clarithromycin (6.00g, 7.87mmol) and reaction at room temperature for 48 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a light yellow crude product (11.14 g).

MS(ESI⁺，m/e)：1022.59[M+H]⁺

Example 319-cinnamoylhydrazone clarithromycin (Compound 7)

Dissolving the crude 2' -O-cinnamoyl-9-cinnamoylhydrazone clarithromycin (middle 19, 1.20g and 0.85mmol) in methanol (6mL), heating and refluxing for 16h, evaporating to dryness under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate, petroleum ether and diethylamine (5: 8: O.6) ], thereby obtaining yellow bubbles (0.67g, wherein the total yield of the compound 7 from the intermediate 2 to the intermediate 19 is 88.7%).

MS(ESI⁺，m/e)：892.45[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.35(s, 1H, 9 ═ N-H), 7.25-7.80(m, 5H, 9-position benzene ring), 7.17(d, J ═ 15.6Hz, 1H, 9 ═ CH-Ph), 6.34(d, J ═ 15.6Hz, 1H, 9-CO-CH ═ c)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 177.85(9-NH-CO-), 167.06(9 > C ═ N-), 144.06(9 ═ CH-Ph), 117.41(9-CO-CH ═ H), 103.14-135.83 (6 carbons on the benzene ring of the side chain at position 9)

Example 322' -O-cinnamoyl-3-O-decoladinose-3-hydroxy-9-cinnamoylhydrazone clarithromycin (intermediate 20)

Dissolving the crude 2' -O-cinnamoyl-9-cinnamoylhydrazone clarithromycin (19.00 g in the middle and 1.41mmol in 1% ethanol solution (20mL), reacting at normal temperature for 24h, adding water and dichloromethane into the filtrate, adjusting the pH value to 9.7 with 3N NaOH, separating out an organic phase, extracting the aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to obtain a yellow foamy substance (1.68 g).

MS(ESI⁺，m/e)：864.49[M+H]⁺

Example 332' -O-cinnamoyl-3-O-decoladinose-3-oxo-9-cinnamoylhydrazone clarithromycin (intermediate 21)

The crude 2' -O-cinnamoyl-3-O-decladinose-3-hydroxy-9-cinnamoylhydrazone clarithromycin (intermediate 20, 1.61g, 1.41mmol) and EDC. HCl (2.47g, 12.50mmol) were dissolved in anhydrous dichloromethane (16mL), DMSO (2.46mL, 33.58mmol) was added, a solution of TFA. Py (1.19g, 6.16mmol) in dichloromethane, N was slowly added dropwise₂Reacting at normal temperature for 0.5h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a yellow sticky solid (1.61 g).

MS(ESI⁺，m/e)：862.48[M+H]⁺

Example 343-O-decoladinose-3-oxo-9-cinnamoylhydrazone clarithromycin (Compound 16)

Dissolving the crude 2' -O-cinnamoyl-3-O-decoladinose-3-oxo-9-cinnamoylhydrazone clarithromycin (intermediate 21, 1.61g and 1.41mmol) in methanol (8mL), heating and refluxing for 3h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography separation [ ethyl acetate, petroleum ether and diethylamine (5: 8: 0.6) ], thus obtaining a yellow bubble (0.35g, from the intermediate 2 to the intermediates 19, 20 and 21, and finally to the compound 16 with a total yield of 33.9%)

MS(ESI⁺，m/e)：732.41[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 11.32(s, 1H, 9 ═ N-H), 7.73(t, J ═ 17.6Hz, 1H, 9-position phenyl ring 4-H), 7.33-7.59(m, 4H, 9-position phenyl ring 2, 3, 5, 6-H, 1H, 9 ═ CH-Ph), 6.34(d, J ═ 15.6Hz, 1H, 9-CO-CH ═ H)

¹³CNMR(400MHz，CDCl₃)δ(ppm)：202.85(3＞C＝O)，167.06(9-NH-CO-)，161.91(9＞C＝N-)，142.14 (9-CH-Ph), 117.25 (9-CO-CH), 120.23-135.81 (6 carbons on the phenyl ring of the 9-position side chain)

Example 352' -O- (2-Thiophenylacetyl) -9- (2-thiopheneacetyl) hydrazone clarithromycin (intermediate 22)

2-Thiopheneacetic acid (2.80g, 19.69mmol) and DCC (4.22g, 20.45mmol) were dissolved in dichloromethane (30mL) and reacted at room temperature for 30min, and 9-hydrazone clarithromycin (3.00g, 3.94mmol) was added and reacted at room temperature for 24 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a brown crude product (5.19 g).

MS(ESI⁺，m/e)：1010.50[M+H]⁺

Example 369- (2-Thiopheneacetyl) hydrazone clarithromycin (Compound 8)

Dissolving the crude 2' -O- (2-thiopheneacetyl) -9- (2-thiopheneacetyl) hydrazone clarithromycin (intermediate 22, 1.60g, 1.21mmol) in methanol (8mL), heating and refluxing for 2h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate, petroleum ether and diethylamine (5: 8: 0.6) ], thereby obtaining a brown bubble (0.61g, from the intermediate 2 to the intermediate 22, and finally obtaining the compound 8, wherein the total yield is 57.0%).

MS(ESI⁺，m/e)：886.31[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.57(s, 1H, 9 ═ N-NH), 6.92 to 7.26(m, 3H, 9 thiophene ring)，4.46(s，2H，9-CO-CH₂-)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 171.41(9-NH-CO-), 168.21(9 > C ═ N-), 124.72-135.57 (4 carbons on the thiophene ring at the 9-position side chain), 33.88 (alkyl carbon between the acyl structure at the 9-position and the thiophene ring)

Example 372' -O- (3-Indoleacetyl) -9- (3-Indoleacetyl) hydrazone clarithromycin (intermediate 23)

3-indoleacetic acid (3.45g, 19.69mmol) and DCC (4.22g, 20.45mmol) were dissolved in dichloromethane (30mL), the solution was cloudy, reacted at room temperature for 30min, added 9-hydrazone clarithromycin (3.00g, 3.94mmol), and reacted at room temperature for 18 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3M NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a brown yellow crude product (4.45 g).

MS(ESI⁺，m/e)：1076.61[M+H]⁺

Example 389- (3-Indolylacetyl) hydrazone clarithromycin (Compound 9)

Dissolving the crude 2' -O- (3-indolacetyl) -9- (3-indolacetyl) hydrazone clarithromycin (intermediate 23, 1.30g and 1.15mmol) in methanol (6.50mL), heating and refluxing for 5h, evaporating under reduced pressure, adding water and dichloromethane, adjusting the pH value to 9.7 by 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, evaporating under reduced pressure, and performing column chromatography [ ethyl acetate: petroleum ether: diethylamine (7: 5: 0.55) ], thereby obtaining a brown yellow foam (0.41g, wherein the total yield of the compound 9 from the intermediate 2 to the intermediate 23 is 39.0%).

MS(ESI⁺，m/e)：919.51[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.71(s, 1H, 9-indole ring NH), 8.48(s, 1H, 9 ═ N-NH), 7.64(d, 1H, 4-H of 9-indole ring), 7.33(d, 1H, 7-H of 9-indole ring), 7.11-7.20(m, 3H, 2, 5, 6-H of 9-indole ring), 3.50(s, 2H, 9-CO-CH)₂-)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 171.59(9-NH-CO-), 166.57(9 > C ═ N-), 121.81-136.59 (8 carbons on the indole ring at the 9-position side chain), 29.78 (alkyl carbon between the acyl structure at the 9-position and the indole ring)

Example 392' -O- (3-Indolbutyryl) -9- (3-Indolbutyryl) hydrazone clarithromycin (intermediate 24)

3-indolebutyric acid (4.00g, 19.68mmol) and DCC (4.22g, 20.45mmol) were dissolved in dichloromethane (30mL) and reacted at room temperature for 30min, and 9-hydrazone clarithromycin (3.00g, 3.94mmol) was added and reacted at room temperature for 45 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a brown yellow solid crude product (6.05 g).

MS(ESI⁺，m/e)：1132.67[M+H]⁺

Example 409- (3-Indolbutyryl) hydrazone clarithromycin (Compound 10)

Dissolving the crude 2' -O- (3-indolebutyryl) -9- (3-indolebutyryl) hydrazone clarithromycin (intermediate 24, 1.40g and 0.91mmol) in methanol (7mL), heating and refluxing for 8h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating an organic phase, extracting an aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline solution, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and performing column chromatography [ ethyl acetate: petroleum ether: diethylamine (5: 6: 0.5) ], thus obtaining yellow bubbles (0.47g, from which the total yield of the intermediate 2, the intermediate 24 and the compound 10 is 54.4%)

MS(ESI⁺，m/e)：947.51[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 8.56(s, 1H, 9-indole ring NH)8.45(s, 1H, 9 ═ N-NH), 7.59(d, J ═ 7.6Hz, 1H, 4-H at 9-indole ring), 7.30(d, J ═ 8.4Hz, 1H, 7-H at 9-indole ring), 6.99-7.16(m, 3H, 2, 5, 6-H at 9-indole ring), 2.85(t, J ═ 7.0Hz, 2H, 9-CH)₂-indole ring), 2.0-2.63(m, 4H, two CH's adjacent to the acyl structure at position 9)₂)

¹³CNMR(400MHz，CDCl₃) δ (ppm): 174.92(9-NH-CO-), 166.46(9 > C ═ N-), 111.02-136.56 (8 carbons on the indole ring at the 9-position side chain), 35.08, 31.97, 24.56 (3 alkyl carbons between the acyl structure at the 9-position and the indole ring)

Example 412' -O-isobutyryl-9-isobutyrylhydrazone clarithromycin (intermediate 25)

Isobutyric acid (2.33mL, 25mmol) and DCC (4.38g, 21.25mmol) were dissolved in dichloromethane (38mL), the solution was cloudy, reacted at room temperature for 30min, added 9-hydrazone clarithromycin (3.81g, 5.00mmol) and reacted at room temperature for 72 h. Filtering, removing insoluble substances, adding water into the filtrate, adjusting pH to 9.7 with 3N NaOH, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white crude product (4.10 g).

MS(ESI⁺，m/e)：902.59[M+H]⁺

Example 422' -O-isobutyryl-3-O-decladinose-3-hydroxy-9-isobutyrylhydrazone clarithromycin (intermediate 26)

Dissolving the 2' -O-isobutyryl-9-isobutyrylhydrazone clarithromycin (25, 4.10g and 5.00mmol in the middle) in 1% hydrochloric acid ethanol solution (20mL), reacting at normal temperature for 24h, adding water, dichloromethane and 3N NaOH to adjust the pH value to 9.7, separating out an organic phase, extracting the aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (3.74 g).

MS(ESI⁺，m/e)：744.49[M+H]⁺

Example 432' -O-isobutyryl-3-O-decladinose-3-oxo-9-isobutyrylhydrazone clarithromycin (intermediate 27)

The crude 2' -O-isobutyryl-3-O-decladinose-3-hydroxy-9-isobutyrylhydrazone clarithromycin (intermediate 26, 3.24g, 4.33mmol) and EDC. HCl (4.33g, 22.60mmol) were dissolved in anhydrous dichloromethane (30mL), DMSO (3.54mL, 45.30mmol) was added, a solution of TFA. Py (2.61g, 22.6mmol) in dichloromethane, N was slowly added dropwise₂Reacting at normal temperature for 24h under protection, adding water, stirring for 10min, separating out an organic phase, extracting an aqueous phase with dichloromethane for three times, combining the organic phases, washing with water, washing with saturated salt water, drying with anhydrous sodium carbonate, filtering, and evaporating the filtrate under reduced pressure to dryness to obtain a white solid (2.22 g).

MS(ESI⁺，m/e)：742.48[M+H]⁺

Example 443-O-decladinose-3-oxo-9-isobutyrylhydrazone clarithromycin (Compound 17)

Dissolving the crude 2' -O-isobutyryl-3-O-decladinose-3-oxo-9-isobutyrylhydrazone clarithromycin (intermediate 27, 2.22g and 4.33mmol) in methanol (20mL), heating and refluxing for 3h, evaporating under reduced pressure, adding water and dichloromethane, adjusting pH to 9.7 with 3N NaOH, stirring for 10min, separating out the organic phase, extracting the aqueous phase with dichloromethane three times, combining the organic phases, washing with water, washing with saturated saline, drying with anhydrous sodium carbonate, filtering, evaporating the filtrate under reduced pressure, and separating by column chromatography [ methanol: chloroform (1: 10) ], to obtain white bubbles (0.45g, from intermediate 2 to intermediate 25, 26 and 27, and finally to compound 17, the total yield is 15.5%).

MS(ESI⁺，m/e)：672.44[M+H]⁺

Example 459-Hydrazone erythromycin thiocyanate A (intermediate 25)

Erythromycin thiocyanate A (20g, 0.0252mol) was dissolved in methanol (20mL), 85% hydrazine hydrate (2.2mL, 0.0378mol) was added, the mixture was refluxed for 18 hours, cooled, a solid precipitated, filtered, and the filter cake was washed with water to give a white solid (13.3g, 65.0%).

9-Hydrazone erythromycin A (intermediate 26)

9-Hydrazone erythromycin thiocyanate A (intermediate 25) (4.00g, 0.0050mol) was dissolved in methylene chloride (90mL) -water (80mL), adjusted to pH9.7 with 3mol/L sodium hydroxide solution, the organic phase was separated, washed with water, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure to give a white foam (3.352g, 90.4%).

EXAMPLE 46N- [4- (2, 5-Dichlorophenoxy) -2-butenyl ] -9-hydrazone erythromycin A (Compound 18)

9-Hydrazone erythromycin A (intermediate 26) (2.00g, 0.0025mol) was dissolved in DMF (20mL), anhydrous potassium carbonate (0.69g, 0.0050mol)4- (2, 5-dichlorophenoxy) -2-butene-1-bromo (0.96g, 0.0033mol) was added and stirred at room temperature for 16h, then water (50mL) and dichloromethane (30mL) were added, the layers were allowed to stand to separate out the organic phase, the aqueous phase was extracted with dichloromethane, the organic phases were combined, washed with water, dried over anhydrous sodium sulfate, filtered, the filtrate was evaporated to dryness under reduced pressure, and column chromatography [ eluent: methanol-dichloromethane (1: 13) ] was performed to obtain a white solid (0.604g, 25.1%).

MS(ESI⁺，m/z)：962[M+H]⁺

¹HNMR(400MHz，CDCl₃) δ (ppm): 7.47(d, J ═ 8.4, 1H, 9-phenyl ring 3-H), 7.26(d, J ═ 2Hz, 1H, 9-phenyl ring 6-H), 7.05(dd, 1H, 9-phenyl ring 4-H), 6.09- δ 6.32(m, 2H, 9-CH ═ CH-), 5.06-5.10(dd, 1H, 1' -CH-).

¹³CNMR(400MHz，CDCl₃) δ (ppm): 174.7(1-CO), 165.7(9-C ═ N-), 154.1 (9-phenyl ring 1-C), 137.8(3-CH ═ CH ═ 132.5 (9-phenyl ring 5-C), 131.1 (9-phenyl ring 3-C), 121.8 (9-phenyl ring 4-C), 120.8 (9-phenyl ring 6-C), 114.8 (9-phenyl ring 6-C), 100.9(1 '-CH), 95.9(3-O-CH), 82.7(5-CH), 78.8(3-CH), 77.3(4 ″ -CH), 76.1(13-CH), 74.1(12-C), 73.8(6-C), 71.1 (2' -CH), 70.7(5 ″ -CH), 68.5 (3-O-CH), and the like₂)，65.7(3′-CH)，65.2(3-CH＝CH-CH₂)，44.1(2-CH)，38.0(7-CH₂)，35.1(4′-CH₂)，21.1(6-CH₃)，21.0(13-CH₂-CH₃)，18.7(8-CH₃)，16.2(12-CH₃)，14.6(10-CH₃)，10.6(13-CH₂-CH₃)，9.6(4-CH₃)。

Example 47N- (2-diethylaminoethyl) -9-hydrazone erythromycin A (Compound 19)

Dissolving 9-hydrazone erythromycin A (intermediate 26) (1.30g, 0.0016mol) in absolute ethanol (7mL), adding diethylaminoethyl chloride hydrochloride (0.3053g, 0.0018mol), heating under reflux and stirring for 7h, adding dichloromethane (25mL) and water (20mL), adjusting pH to 9.3, standing for layering, separating out the organic phase, extracting the aqueous phase with dichloromethane, combining the organic phases, washing with water, drying with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure, and separating by column chromatography [ eluent: ethyl acetate-petroleum ether-diethylamine (5: 16: 1) ] to obtain white foam, and separating by column chromatography [ eluent: methanol-dichloromethane-ammonia water (1: 25: 0.5) ] to obtain white foam (0.055g, 4.1%).

MS(ESI⁺，m/z)：847[M+H]⁺

¹HNMR(400MHz，DMSO)δ(ppm)：5.57(s，1H，9-N-NH-)，5.08-5.11(dd，1H，1′-CH-)，2.32-2.64(m，8H，9-NH-CH ₂ CH ₂N(CH ₂CH₃)₂)。

¹³CNMR(400MHz，DMSO)δ(ppm)：174.7(1-CO)，δ165.7(9-C＝N-)，102.9(1′-CH)，96.4(3-O-CH)，83.7(5-CH)，80.4(3-CH)，78.1(4″-CH)，77.3(12-C)，77.0(6-C)，76.8(13-CH)，76.7(5″-CH)，71.5(2′-CH)，65.5(3′-CH)，51.6(9-NH-CH₂ CH₂N)，48.0(9-NH-CH₂CH₂N)，45.6(9-NH-CH₂CH₂N(CH ₂CH₃)₂)，44.7(2-CH)，40.3(3′-N(CH₃)₂)，38.7(7-CH₂)，35.2(6″-CH)，32.6(4-CH)，28.9(4′-CH₂)，21.5(6-CH₃)，21.1(13-CH₂-CH₃)，18.6(8-CH₃)，16.0(12-CH₃)，14.3(10-CH₃)，10.5(13-CH₂-CH₃)，9.2(4-CH₃)。

Effects of the embodiment

And (3) testing antibacterial activity:

and (3) carrying out in-vitro antibacterial activity test on the synthesized compounds 1-19, dissolving the samples with absolute ethyl alcohol, diluting the samples with sterile water to 125 mu g/mL, and then sequentially carrying out double dilution. 20 strains G⁺And G^-The test strains were inoculated in broth and cultured overnight at 37 ℃. Diluting with agar plate, and quantitatively inoculating 10 dots by using multi-dot inoculating instrument⁵And (4) CFU. After the strains are inoculated, the strains are placed in an incubator at 37 ℃ for culturing for 18h to observe the result, and the minimum inhibitory concentration (MIC value) of the compound to the tested bacteria is obtained.

The test bacteria include G⁺6 strains of the strain: staphylococcus aureus 26003, Staphylococcus epidermidis 26069, Staphylococcus albus 26101,Pneumococci 31002, enterococcus 32220, and streptococcus c 32206, G^-14 strains: escherichia coli 44102, Pseudomonas aeruginosa 10124, Klebsiella pneumoniae 46101, Salmonella typhi 50097, Aerobacter aerogenes 45102, Citrobacter 48017, Proteus vulgaris 49085, Proteus mirabilis 49005, Proteus morganii 49086, Salmonella enteritidis 50041, Serratia marcescens 41002, Shigella sonnei 51081, Shigella boydii 51313 and Shigella flexneri 51573.

The test results are shown in tables 3 and 4.

TABLE 3

TABLE 4

And (4) conclusion: the synthesized 9-substituted acylhydrazone clarithromycin derivative (compound 1-10) has strong antibacterial activity on staphylococcus albus, diplococcus pneumoniae and enterococcus, the compound 1 and the compound 9 have strong antibacterial activity on staphylococcus aureus, and the compound 1, 2, 3, 4, 5, 6, 8, 9 and 10 have strong antibacterial activity on streptococcus type C. Wherein the antibacterial activity of the compounds 1, 4 and 9 to staphylococcus aureus is better than that of azithromycin, and the antibacterial activity of the compounds 1 to 10 to enterococcus is better than that of azithromycin.

The synthesized 3-O-descladinose-3-oxo-9-substituted acylhydrazone clarithromycin derivative (compound 11-17) has strong antibacterial activity against staphylococcus aureus, staphylococcus albus, pneumococcus and streptococcus type C, and part of the compound (compound 11, 12, 16, 17) has strong antibacterial activity against enterococcus. The antibacterial activity of all the compounds on staphylococcus aureus is superior to that of azithromycin (compounds 11-17). There were 4 compounds (compounds 11, 12, 14, 16) with better activity against streptococcus type c than azithromycin; the antibacterial activity of 4 compounds (compounds 11, 12, 16, 17) against enterococcus was superior to that of azithromycin.

Claims

1. Macrolide compounds shown as a formula I;

wherein R is C₁～C₃Alkyl, or H; r^aIs composed of

In the 3-position

Represents a single bond or a double bond, in which case R represents a double bond^bIn the absence of, which is a single bond, R^bIs composed of

R^cIs substituted or unsubstituted C₁～C₃Alkyl, substituted or unsubstituted C₆～C₁₀Aryl, or substituted or unsubstituted C₂～C₄An alkenyl group; wherein, substituted C₆～C₁₀The substituent in the aryl is nitro or halogen, and the substituent in the aryl is ortho, para or meta; substituted C₁～C₃The substituent in the alkyl group being C₆～C₁₀Aryl, or C₄～C₈Heteroaryl, wherein the heteroatom is N, O or S; substituted C₂～C₄The substituent in the alkenyl group is C₆～C₁₀An aryl group;

X¹and X²Independently is halogen;

R^dand R^eIndependently is C₁～C₃An alkyl group;

n is 1 to 3.

2. Macrolide compound according to claim 1, characterized in that: when R is^cIs substituted or unsubstituted C₁～C₃When alkyl, said C₁～C₃Alkyl is methyl, ethyl or isopropyl.

3. Macrolide compound according to claim 1, characterized in that: r^cIn (b), said substituted C₆～C₁₀When the substituent in the aryl is halogen, the halogen is Cl.

4. Macrolide compound according to claim 1, characterized in that: r^cIn (1), substituted C₁～C₃The substituent in the alkyl group being C₄～C₈When it is heteroaryl, said C₄～C₈Heteroaryl is 2-thienyl or 3-indolyl.

5. Macrolide compound according to claim 1, characterized in that: when X is present¹And X²When independently halogen, the halogen is fluorine, chlorine, bromine or iodine.

6. Macrolide compound according to claim 1, characterized in that: said X¹And X²The positions of (a) are 2 'and 5'.

7. Macrolide compound according to claim 1, characterized in that: said X¹And X²The same is true.

8. Macrolide compound according to claim 1, characterized in that: when R is^dAnd R^eIndependently is C₁～C₃When alkyl, said C₁～C₃The alkyl group is ethyl.

9. Macrolide compound according to claim 1, characterized in that: and n is 2.

10. Macrolide compound according to any of claims 1 to 9, characterized in that: the compound I has any one of the following structures:

wherein Ra and Rc are as defined in any one of claims 1 to 9.

11. Macrolide compound according to any of claims 1 to 9, characterized in that: the compound I is any one of the following compounds:

in Ia, R^cIs methyl, phenyl, p-nitrophenyl, p-chlorophenyl, benzyl, phenethyl, styryl, thiophene-2-methyl, indole-3-propyl;

in Ic, R^aIs composed of

12. A process for producing a macrolide compound according to any one of claims 1 to 9, wherein: it is any one of the following methods:

(I) when the object compound I, R^aIs composed of

When R is C₁～C₃When in alkyl, the compound II is subjected to a reaction of removing the acyl protecting group of the hydroxyl;

R^c1is R^cOr substituents on acyl protecting groups conventional in the art, each group being as defined in any one of claims 1 to 9 unless otherwise specified;

(II) when the object compound I, R^aIs composed of

wherein X is a leaving group which is conventional in such nucleophilic substitution reactions in the art, and the definitions of each group are as described in any one of claims 1 to 9 unless otherwise specified.

13. An intermediate compound IIa, IIb or IIIb for the preparation of a compound I according to any one of claims 1 to 9:

wherein each group is as defined in any one of claims 1 to 9.

14. The use of a compound I according to any one of claims 1 to 9 for the preparation of an antibiotic medicament.