CN112891334B

CN112891334B - Application of caffeic acid derivatives in preparation of medicine for treating gonorrhea

Info

Publication number: CN112891334B
Application number: CN202110070169.4A
Authority: CN
Inventors: 斯戴·樊德冯; 西蒙·杜特怀勒; 张江临; 杨帆; 严景; 孙雨吉
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2021-01-19
Filing date: 2021-01-19
Publication date: 2022-05-03
Anticipated expiration: 2041-01-19
Also published as: CN112891334A

Abstract

The invention discloses an application of caffeic acid derivatives in preparing a medicine for treating gonorrhea, wherein the caffeic acid derivatives are shown as a formula (I). The caffeic acid derivative compound shown in the formula (2) has extremely strong bactericidal activity on gonococcus in vitro, and shows that the caffeic acid derivative compound has wide application prospect in preparing medicaments for treating infectious diseases caused by gonococcus.

Description

Application of caffeic acid derivatives in preparation of medicine for treating gonorrhea

Technical Field

The invention relates to the field of antibacterial drugs, in particular to application of caffeic acid derivatives in preparing drugs for treating gonorrhea.

Background

Humans are the only hosts of gonococci, and sexually transmitted diseases are usually caused after gonococci infection, and currently, gonococci have developed drug resistance to antibiotics which have been mostly used as empirical drugs, such as penicillin, tetracycline, sulfonamides, trimethoprim, (fluoro) quinolones, macrolides, and the like.

Ceftriaxone is an internationally recognized first-line experience medicine, the drug resistance rate of ceftriaxone in U.S. in 2013-2014 is 0.1%, the drug resistance rate in Europe in 2014 is 0.25%, and the drug resistance rate of ceftriaxone in Australia in 2014 is 0.6%, however, the low sensitivity rate of ceftriaxone in 2013-2015 in China is up to 10.8%, so the status of ceftriaxone as the first-line medicine is knocked to the police, but no good substitute medicine is put into the market at present, and therefore new antibacterial medicines are urgently needed to prevent the situation of no medicine and medical treatment appearing in the future.

Phytochemicals are a large group of naturally occurring compounds, which are widely present in natural plants, and are classified into several groups, such as alkaloids, terpenoids, and polyphenols, according to the structural differences of these natural compounds.

Through research, some of the compounds can protect organisms from free radicals, viruses, bacteria and fungi, and can show excellent bactericidal and bacteriostatic capacities by inhibiting adhesion and infection of bacteria, inhibiting expression of bacterial virulence factors, damaging or destroying cell membranes of the bacteria, inhibiting formation of bacterial biofilms and the like, for example, berberine can effectively inhibit adhesion capacity and biofilm formation of staphylococcus epidermidis, and 6-gingerol derived from ginger pungent oil can inhibit production of virulence factors of pseudomonas aeruginosa and biofilm formation.

There are many similar natural compounds with antibacterial and bacteriostatic capabilities, and it is expected that botanical chemicals have great potential for developing new effective, safe, and inexpensive antimicrobial agents against the impending current state of gonococcal drug resistance.

Chinese patent application publication No. CN102795953B discloses a synthesis method and use of caffeic acid amide derivatives from caffeic acid and sulfonamides as raw materials, wherein the caffeic acid amide derivatives synthesized by the method have inhibitory effects on pneumococcus, escherichia coli, pseudomonas aeruginosa, dysentery bacillus, salmonella and escherichia coli.

The Chinese patent application with the publication number of CN104823974A discloses the application of caffeic acid alkyl ester to 3 gram-positive bacteria, 2 gram-negative bacteria and 1 fungus.

However, the prior art has not reported any way as to whether caffeic acid derivatives can be used to treat gonorrhea.

Disclosure of Invention

The invention aims to provide novel caffeic acid phenethyl ester and derivatives thereof which can be used for treating infectious diseases caused by gonococcus, and elucidate the application of the natural compound in the infectious diseases caused by gonococcus. The natural compound has extremely high antibacterial activity on gonococcus and wide application prospect.

The invention adopts the following specific technical scheme:

the application of the compound of the formula (I) in preparing the medicine for treating gonorrhea,

wherein R is₁Is H or OH;

L₁is C_1～3Alkylene or a single bond;

R₂is H or carboxyl;

R₃is phenyl or C_2～5Alkenyl, said phenyl or C_2～5Alkenyl is optionally substituted by 1,2 or 3R_aSubstitution; r_aIs H, F, Cl, Br, I, CH₃、OH、NH₂Or CN.

Said C_1～3Alkylene is-CH₂-、-CH₂CH₂-、-CH₂ CH₂CH₂-or-CH (CH)₃)CH₂-。

Said R₃Is phenyl or

Said phenyl group or

Optionally substituted with 1,2 or 3 halogens.

Said R₁Is OH; l is₁is-CH₂-or a single bond; r₂Is H; r₃Is phenyl or

Preferably, the compound of formula (I) is represented by the compounds of formulae (2) to (7):

the compound of formula (I) is further preferably a compound of formulae (2) to (4), wherein the compounds of formulae (2) to (4) show strong anti-gonococcal activity, and the minimum inhibitory concentration is 32 mu M and 64 mu M respectively.

The gonorrhea is an infectious disease caused by gonococci; the gonococcus is resistant to ceftriaxone, cefixime, azithromycin, penicillin, ciprofloxacin or tetracycline.

The infection is a urogenital, rectal, throat or eye infection.

The pharmaceutical composition comprises the compound shown in the formula (I) and pharmaceutically acceptable auxiliary materials, and is prepared into a pharmaceutically common pharmaceutical preparation which is an emulsion, a gel, a capsule, a suppository, a tablet, a solvent or a suspension.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent, and may include variations of deuterium and hydrogen, so long as the valency of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., ═ O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemical realizability.

As used herein, the term "alkenyl" by itself or as part of another substituent refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon double bond. The alkenyl group having one double bond may be represented by-C_nH_2n-1Having two double bonds and is represented by-C_nH_2n-3. When one alkenyl group is preceded by a modification of the number of carbon atoms, e.g. C_2-8When it means that the alkenyl group has 2 to 8 carbon atoms. For example, C_2-8Examples of the alkenyl group may include vinyl, allyl, 1, 2-butenyl, 2, 3-butenyl, butadienyl and the like.

Compared with the prior art, the invention has the following beneficial effects:

(1) as a novel antibacterial compound, compared with the existing antibiotics and derivatives thereof, the compound is sensitive to drug-resistant bacteria.

(2) The compound is used as a novel antibacterial compound, is not easy to form drug resistance, and is economical and practical.

Drawings

FIG. 1 is a graph of the time-kill activity of the compound of formula (2) and penicillin on gonococci.

FIG. 2 is a graph showing the minimal inhibitory concentration profile of the compound of formula (2) against a clinical isolate of gonococci.

Detailed Description

Specific embodiments of the present invention are described in detail in the following examples. These are illustrative of the invention and do not limit the scope of the invention. All reagents were commercially available unless otherwise indicated.

Example 1

The in vitro activity of the compounds of the invention can be assessed according to the EUCAST test Standard for drug sensitivity in the European Union (EUCAST; www.eucast.org).

According to the test standard of the drug sensitivity test of the European Union, the Minimum Inhibitory Concentrations (MIC) of different pathogenic bacteria to the compounds of the formulae (2) to (7) of the present invention are detected by an agar plate dilution method. The bacteria were recovered from the ultra-low temperature refrigerator and inoculated on GC agar plates containing 1% Vitox, 37 ℃ and 5% CO₂Culturing in an inverted manner for 14-18 h, scraping bacteria to prepare bacterial suspension (OD)₆₀₀0.01), 5. mu.l of the bacterial solution was aspirated by a micro-applicator and spotted on a plate containing the compound of the present invention at each concentration, air-dried, and subjected to 5% CO at 37 ℃ to air-dry₂Culturing for 24-48h under the condition, and observing and recording the MIC value. The MIC values of the compounds of the invention against different pathogenic bacteria are shown in table 1.

TABLE 1 minimum inhibitory concentration (MIC, μ M) of the compounds of the invention against different pathogenic bacteria

Example 2

Neisseria gonorrhoeae (ATCC 49226) cultured overnight was prepared as OD in GC liquid medium₆₀₀To a bacterial suspension of 1, a predetermined amount of the bacterial suspension was added to 12mL of GC liquid medium containing 1% vitox (10)⁷CFU/mL). After culturing the cells in a shaker at 37 ℃ and 200rpm/min for 1h, the compound of formula (2) was added, and penicillin was used as a positive control.

At different time points, samples were taken and diluted to the appropriate concentration, 100. mu.L of diluted bacterial solution was pipetted and spread on a GC agarose plate, which was placed at 37 ℃ with 5% CO₂For 24-48h, observing and recording the number of Clones (CFU).

The results are shown in FIG. 1, and are expressed as mean. + -. SD of three biological replicates, with the dotted line representing the lower limit of detection. In contrast to penicillin, the compound of formula (2) was effective in rapidly killing gonococci at concentrations of 4 × MIC (4 × 32 μ M) and 2 × MIC (2 × 32 μ M), and also at 1 × MIC (1 × 32 μ M), and 1/2 × MIC (1/2 μ M) (2) was still able to inhibit the growth of gonococci to some extent, indicating that the compound of formula (2) had good bactericidal activity against gonococci.

Example 3

The Minimal Inhibitory Concentration (MIC) of a compound of formula (2) of the present invention was determined by agar plate dilution method for 100 clinical isolates of gonococci (n 100) containing drug-resistant gonococci to ceftriaxone, cefixime, azithromycin, penicillin, ciprofloxacin and tetracycline, according to eu drug susceptibility test standards. The bacteria were recovered from the ultra-low temperature refrigerator and inoculated on GC agar plates containing 1% Vitox, 37 ℃ and 5% CO₂Culturing in an inverted manner for 14-18 h, scraping bacteria to prepare bacterial suspension (OD)₆₀₀0.01), 5. mu.l of the bacterial solution was aspirated by a microsyringe and spotted on a medium containing the compound of the present invention at each concentrationDrying the mixture in air at 37 deg.C with 5% CO₂Culturing for 24-48h under the condition, and observing and recording the MIC value.

The result is shown in figure 2, the minimum inhibitory concentration of the compound shown in the formula (2) on the gonococcus clinical isolate is distributed between 32 mu M and 64 mu M, and the clinically isolated gonococcus is proved to be sensitive to the compound shown in the formula (2), so that the compound shown in the formula (2) can effectively kill the gonococcus and is beneficial to the application of the compound shown in the formula (2) to the treatment of infectious diseases caused by the gonococcus.

Claims

1. The use of a compound of formulae (2), (3) and (4) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of gonorrhea,

2. the use according to claim 1, wherein the gonorrhea is an infectious disease caused by gonococci; the gonococcus is resistant to ceftriaxone, cefixime, azithromycin, penicillin, ciprofloxacin or tetracycline.

3. The use according to claim 2, wherein the infection is a urogenital, rectal, throat or eye infection.

4. The application of a pharmaceutical composition in preparing a medicament for treating gonorrhea is characterized in that the pharmaceutical composition comprises at least one of the compounds of the formulae (2), (3) and (4) as defined in claim 1 or pharmaceutically acceptable salts thereof and pharmaceutically acceptable auxiliary materials to prepare a pharmaceutically common pharmaceutical preparation, wherein the pharmaceutical preparation is an emulsion, a gel, a capsule, a suppository, a tablet or a suspension.

5. The use of the pharmaceutical composition according to claim 4 for the preparation of a medicament for the treatment of gonorrhea, wherein said gonorrhea is an infectious disease caused by gonococci; the gonococcus is resistant to ceftriaxone, cefixime, azithromycin, penicillin, ciprofloxacin or tetracycline.