Application of isoliquiritigenin and derivatives thereof in preparation of anti-gonococcus drugs

Abstract

The invention provides application of isoliquiritigenin and derivatives thereof in preparation of anti-gonococcus drugs, and relates to the technical field of biology. The isoliquiritigenin and the derivatives thereof can inhibit gonococci, and part of the isoliquiritigenin derivatives and antibiotics are combined to inhibit gonococci, so that a synergistic effect is achieved, and the problem of drug resistance of gonococci to existing drugs is relieved.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to application of isoliquiritigenin and its derivatives in the preparation of anti-gonococcal drugs. Background Art

[0002] The following statements merely provide background information related to the present disclosure and do not necessarily constitute prior art.

[0003] Neisseria gonorrhoeae (N.gonorrhoeae) is a gram-negative diplococcus of the genus Neisseria and the pathogen of gonorrhea. The bacteria are arranged in two balls and are kidney-shaped. They have no spores and flagella, but have pili. Gonorrhea mainly colonizes the urogenital tract mucosa of human hosts and can cause suppurative infections of the urogenital system. If not effectively treated, ascending infections can cause a variety of complications, such as epididymitis, cervicitis, pelvic inflammatory disease, salpingitis, ectopic pregnancy and infertility. It can also promote the infection and spread of HIV, herpes simplex virus, Chlamydia trachomatis and syphilis. Neisseria gonorrhoeae is very easy to acquire drug resistance and has a high incidence rate. In 2016, the World Health Organization (WHO) estimated that the incidence of gonorrhea in adults aged 15-49 was approximately 86.9 million per year. In 2020, the global incidence rate was 19 cases per 1,000 women and 23 cases per 1,000 men.

[0004] There is no effective vaccine for gonorrhea. Antibiotics are currently the only most effective treatment for gonorrhea. However, gonococci have developed serious resistance to antibiotics and are now resistant to almost all antibiotics. Since the first introduction of sulfonamides to treat gonorrhea in the 1930s, gonococcal resistance to antibiotics has increased rapidly, and serious resistance has emerged to the once recommended first-line and second-line drugs such as sulfonamides, penicillins, tetracyclines, early macrolides, fluoroquinolones and early-generation cephalosporins. The WHO Gonococcal Resistance Monitoring Project found that gonococcal resistance is spreading globally, especially in Asia, North America, Europe, Latin America and the Caribbean. At present, most countries in the world use ceftriaxone monotherapy as the first-line drug for the treatment of gonorrhea. In 2009, Japan discovered the first strain of high-level resistance to ceftriaxone (super gonococci), which attracted widespread attention around the world. In 2015, a ceftriaxone-resistant strain FC428 (penA-60.001) belonging to sequence type (ST) 1903 was detected in Japan again. Subsequently, this resistant strain spread widely around the world. With the increasing failure of first-line monotherapy, some countries have adopted the combination therapy of adding macrolide drugs azithromycin, but strains with high-level single-drug or double-drug resistance to ceftriaxone or azithromycin have also appeared in many countries. High-level drug resistance of gonococci has become a major public health issue of global concern. Therefore, the first-line treatment drugs for gonorrhea are facing challenges and exhaustion. WHO believes that gonorrhea may once again enter an era where there is no cure for it, and the development of new drugs is imminent.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide the use of isoliquiritigenin and its derivatives in the preparation of anti-gonococcal drugs, and the anti-gonococcal drugs based on the use, so as to alleviate the problem of gonococcal resistance to existing drugs.

[0007] In order to solve the above technical problems, the present invention particularly adopts the following technical solutions:

[0008] In the first aspect, the invention provides the use of isoliquiritigenin and its derivatives in the preparation of anti-gonococcal drugs;

[0009] The isoliquiritigenin and its derivatives include compound 1, compound 2, compound 4, compound 5, compound 6, compound 8, compound 9, compound 10, compound 11, compound 13, compound 14, compound 15, compound 117, compound 20, compound 21, compound 22, compound 24, compound 37, compound 39 and compound 40;

[0010] Wherein compound 1 is isoliquiritigenin, and its structural formula is shown in formula (1);

[0011]

[0012] The structural formulas of Compound 2, Compound 4, Compound 5, Compound 6, Compound 8, Compound 9, Compound 110, Compound 11, Compound 13, Compound 14, Compound 15 and Compound 17 are shown in Formula (I):

[0013]

[0014] Where R1, R2, R3, R4, R5 and R6 are shown in the following table:

[0015] Compound <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R3]]> <![CDATA[R4]]> <![CDATA[R5]]> <![CDATA[R6]]> Compound 1 OH OH H H OH H Compound 2 OH OH H <![CDATA[OCH3]]> OH H Compound 4 H OH H <![CDATA[OCH3]]> OH H Compound 5 H OH H H OH H Compound 6 H OH H H <![CDATA[CF3]]> H Compound 8 OH <![CDATA[OCH3]]> H H OH H Compound 9 OH <![CDATA[OCH3]]> H OH <![CDATA[OCH3]]> H Compound 10 OH <![CDATA[OCH3]]> H <![CDATA[OCH3]]> OH H Compound 11 OH <![CDATA[OCH3]]> H H <![CDATA[CF3]]> H Compound 13 OH <![CDATA[CF3]]> H OH <![CDATA[OCH3]]> H Compound 14 OH OH H OH <![CDATA[OCH3]]> H Compound 15 H Bn H H OH H Compound 17 OH OH <![CDATA[OCH3]]> H H H

[0016] The structural formulas of Compound 20, Compound 21 and Compound 22 are shown in Formula (II):

[0017]

[0018] Where R1, R2 and R3 are shown in the following table:

[0019] The structural formulas of compound 24 and compound 37 are shown in formula (III):

[0020] Compound <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R7]]> Compound 20 OH <![CDATA[OCH3]]> H Compound 21 OH <![CDATA[OCH3]]> OH Compound 22 H Bn H

[0021]

[0022] Where R1, R2, R3, R4, R5 and R6 are shown in the following table:

[0023] Compound <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R3]]> <![CDATA[R4]]> <![CDATA[R5]]> <![CDATA[R6]]> Compound 24 OH OH H H OH H Compound 37 OH <![CDATA[CF3]]> H <![CDATA[OCH3]]> OH H

[0024] The structural formula of compound 39 is shown in formula (39):

[0025]

[0026] The structural formula of compound 40 is shown in formula (40):

[0027] Compound 40:

[0028] In a second aspect, an anti-gonococcal drug is also provided, wherein the anti-gonococcal drug comprises (A) and (B):

[0029] (A) Antibiotics:

[0030] (B) one or more of the isoliquiritigenin and its derivatives described in the first aspect.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] Licorice (Glycyrrhiza) belongs to the perennial leguminous plant and is an extremely important Chinese herbal medicine. Isoliquiritigenin is an important active ingredient in licorice. The pure product is yellow needle-shaped crystals, which are insoluble in water and soluble in less polar solvents such as ether and chloroform. It has a broad spectrum of biological activities, and its anti-inflammatory and anti-cancer activities are the hot spots of current research. The present invention finds that isoliquiritigenin and its derivatives have the effect of inhibiting gonococci, and can inhibit drug-resistant gonococci. The present invention also finds that isoliquiritigenin and its derivatives are used in combination with antibiotics to inhibit gonococci without antagonism, and some isoliquiritigenin derivatives are used in combination with antibiotics to have a synergistic effect. The present invention has achieved new results in anti-gonococcal sensitization based on the combined use of traditional Chinese and Western medicine, and provides new ways and measures for alleviating bacterial resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0034] Figure 1 The lowest FIC value of the combined effect of isoliquiritigenin and its derivatives combined with different antibiotics on gonococcal strains in Example 2;

[0035] Figure 2 This is a heat map of the lowest FIC values ​​of the combined effects of isoliquiritigenin and its derivatives combined with different antibiotics on gonococcal strains in Example 2. DETAILED DESCRIPTION

[0036] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] In the first aspect, the invention provides the use of isoliquiritigenin and its derivatives in the preparation of anti-gonococcal drugs. The anti-gonococcal drugs herein should be understood in the broadest sense, and any preparation having the function of inhibiting gonococci belongs to the anti-gonococcal drugs described herein.

[0038] The isoliquiritigenin and its derivatives include compound 1, compound 2, compound 4, compound 5, compound 6, compound 8, compound 9, compound 10, compound 11, compound 13, compound 14, compound 15, compound 117, compound 20, compound 21, compound 22, compound 24, compound 37, compound 39 and compound 40.

[0039] Compound 1 is isoliquiritigenin, with the chemical formula C 15 H 12 O4, MW: 256.25, structural formula is shown in formula (1);

[0040]

[0041] The remaining compounds are all isoliquiritigenin derivatives. The structural formulas of Compound 2, Compound 4, Compound 5, Compound 6, Compound 8, Compound 9, Compound 10, Compound 11, Compound 13, Compound 14, Compound 15 and Compound 17 are shown in Formula (I):

[0042]

[0043] Where R1, R2, R3, R4, R5 and R6 are shown in the following table:

[0044] Compound <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R3]]> <![CDATA[R4]]> <![CDATA[R5]]> <![CDATA[R6 <!-- 4 -->]]> Compound 2 OH OH H <![CDATA[OCH3]]> OH H Compound 4 H OH H <![CDATA[OCH3]]> OH H Compound 5 H OH H H OH H Compound 6 H OH H H <![CDATA[CF3]]> H Compound 8 OH <![CDATA[OCH3]]> H H OH H Compound 9 OH <![CDATA[OCH3]]> H OH <![CDATA[OCH3]]> H Compound 10 OH <![CDATA[OCH3]]> H <![CDATA[OCH3]]> OH H Compound 11 OH <![CDATA[OCH3]]> H H <![CDATA[CF3]]> H Compound 13 OH <![CDATA[CF3]]> H OH <![CDATA[OCH3]]> H Compound 14 OH OH H OH <![CDATA[OCH3]]> H Compound 15 H Bn H H OH H Compound 17 OH OH <![CDATA[OCH3]]> H H H

[0045] The structural formulas of Compound 20, Compound 21 and Compound 22 are shown in Formula (II):

[0046] Where R1, R2 and R3 are shown in the following table:

[0047] Compound <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R7]]> Compound 20 OH <![CDATA[OCH3]]> H Compound 21 OH <![CDATA[OCH3]]> OH Compound 22 H Bn H

[0048] The structural formulas of compound 24 and compound 37 are shown in formula (III):

[0049]

[0050] Where R1, R2, R3, R4, R5 and R6 are shown in the following table:

[0051]

[0052]

[0053] The structural formula of compound 39 is shown in formula (39):

[0054]

[0055] The structural formula of compound 40 is shown in formula (40):

[0056] Compound 40:

[0057] Bn represents a benzyl group.

[0058] In an optional embodiment, the application includes applying at least one of the following isoliquiritigenin and its derivatives to the preparation of an anti-gonococcal drug: isoliquiritigenin (Compound 1), Compound 2, Compound 5, Compound 8, Compound 9, Compound 10, Compound 13, Compound 14, Compound 15, Compound 20, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40. The above compounds can have an anti-gonococcal effect at a concentration of ≤100 μmol / L.

[0059] In an optional embodiment, the effective concentrations of Compound 1, Compound 2, Compound 5, Compound 8, Compound 9, Compound 10, Compound 13, Compound 14, Compound 15, Compound 20, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40 in the anti-gonococcal drug are independently at least 10 μmol / L, or at least 25 μmol / L, or at least 50 μmol / L, or at least 100 μmol / L.

[0060] In an optional embodiment, the application includes applying at least one of the following isoliquiritigenin derivatives to the preparation of an anti-gonococcal drug: isoliquiritigenin (Compound 1), Compound 9, Compound 10, Compound 15, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40. The above compounds can have an anti-gonococcal effect at a concentration of ≤50 μmol / L.

[0061] In an optional embodiment, the effective concentration of isoliquiritigenin (Compound 1), Compound 9, Compound 10, Compound 15, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40 in the anti-gonococcal drug is independently at least 10 μmol / L, or at least 25 μmol / L, or at least 50 μmol / L.

[0062] In an optional embodiment, the gonococci include drug-resistant gonococci.

[0063] In an optional embodiment, the drug-resistant gonococci include gonococci resistant to at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin and ceftriaxone.

[0064] In an optional embodiment, the application includes applying at least one of the following isoliquiritigenin and its derivatives to the preparation of anti-resistant gonococcal drugs: Compound 1 (isoliquiritigenin), Compound 5, Compound 6, Compound 9, Compound 110, Compound 15, Compound 21, Compound 37, Compound 39 and Compound 40.

[0065] In an optional embodiment, the application includes applying at least one of the following isoliquiritigenin and its derivatives to the preparation of drugs against the above-mentioned drug-resistant gonococci: Compound 1 (isoliquiritigenin), Compound 5, Compound 6, Compound 9, Compound 10, Compound 15, Compound 21, Compound 37, Compound 39 and Compound 40; the antibiotics include at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin, ceftriaxone and cefixime.

[0066] In an optional embodiment, the application includes applying penicillin and at least one of the following isoliquiritigenin derivatives to prepare drugs against drug-resistant gonococci: compound 6, compound 10, compound 21 and compound 39, and the above compounds have a synergistic effect with penicillin.

[0067] In an optional embodiment, the application includes applying tetracycline and at least one of the following isoliquiritigenin derivatives to prepare anti-resistant gonococcal drugs: compound 6, compound 10, compound 21 and compound 39, and the above compounds have a synergistic effect with tetracycline.

[0068] In an optional embodiment, the application includes applying ciprofloxacin and at least one of the following isoliquiritigenin derivatives to prepare an anti-resistant gonococcal drug: compound 6, compound 21 and compound 39, and the above compounds have a synergistic effect with ciprofloxacin.

[0069] In an optional embodiment, the application includes applying azithromycin and at least one of the following isoliquiritigenin derivatives to prepare anti-resistant gonococcal drugs: compound 6, compound 10, compound 15, compound 21 and compound 39, and the above compounds have a synergistic effect with azithromycin.

[0070] In an optional embodiment, the application includes applying ceftriaxone and at least one of the following isoliquiritigenin derivatives to prepare anti-resistant gonococcal drugs: compound 6, compound 10, compound 21 and compound 39, and the above compounds have a synergistic effect with ceftriaxone.

[0071] In an optional embodiment, the application includes applying cefixime and compound 6 (isoliquiritigenin derivative) to the preparation of an anti-drug-resistant gonococcal drug, and compound 6 (isoliquiritigenin derivative) and cefixime have a synergistic effect.

[0072] In a second aspect, an anti-gonococcal drug is provided, comprising (A) and (B):

[0073] (A) Antibiotics:

[0074] (B) one or more of isoliquiritigenin (Compound 1), Compound 2, Compound 4, Compound 5, Compound 6, Compound 8, Compound 9, Compound 10, Compound 11, Compound 13, Compound 14, Compound 15, Compound 17, Compound 20, Compound 21, Compound 22, Compound 24, Compound 37, Compound 39 and Compound 40 described in the first aspect.

[0075] In an optional embodiment, the antibiotic includes at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin, ceftriaxone and cefixime.

[0076] In an optional embodiment, the anti-gonococcal drug comprises at least one of the following isoliquiritigenin and its derivatives: Compound 1 (isoliquiritigenin), Compound 5, Compound 6, Compound 9, Compound 10, Compound 15, Compound 21, Compound 37, Compound 39 and Compound 40.

[0077] In an optional embodiment, the anti-gonococcal drug comprises penicillin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39.

[0078] In an optional embodiment, the anti-gonococcal drug comprises tetracycline and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39.

[0079] In an optional embodiment, the anti-gonococcal drug comprises ciprofloxacin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 21 and Compound 39.

[0080] In an optional embodiment, the anti-gonococcal drug comprises azithromycin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 15, Compound 21 and Compound 39.

[0081] In an optional embodiment, the anti-gonococcal drug comprises ceftriaxone and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39.

[0082] In an optional embodiment, the anti-gonococcal drug comprises cefixime and compound 6 (isoliquiritigenin derivative).

[0083] In an optional embodiment, the anti-gonococcal drug also includes a pharmaceutically acceptable carrier and / or excipient. The acceptable carrier and pharmaceutically acceptable excipient can be selected from any conventional carrier and / or excipient known in the art. Including but not limited to solvents, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, anti-caking agents, flavoring agents, antibacterial agents, suspending agents, coating agents, film-forming agents, aromatics, tackifiers, anti-adhesive agents, antioxidants, antioxidant synergists, chelating agents, pH regulators, adsorbents, plasticizers, surfactants, thickeners, inclusion agents, protective agents, humectants, softeners, absorbents, diluents, release regulators, pressure-sensitive adhesives, hardeners, hollow capsules, one or more of matrix and drug carrier materials.

[0084] In an optional embodiment, the dosage form of the drug can be selected from any acceptable dosage form in the art, including but not limited to injection, nasal preparation, inhalation preparation, spray or oral preparation. The appropriate formulation depends on the selected route of administration, and those skilled in the art can manufacture it according to common sense known in the art, and select suitable excipients.

[0085] The present invention is further described below by means of specific examples. However, it should be understood that these examples are only used for more detailed description and should not be construed as limiting the present invention in any form.

[0086] The structural formulas of the numbered compounds 1 to 40 used in the following examples are as follows, wherein the numbered compound 1 is isoliquiritigenin, and the numbered compounds 2 to 40 are isoliquiritigenin derivatives.

[0087] The structural formula of isoliquiritigenin is shown in formula (1).

[0088]

[0089] The structural formulas of compounds No. 2 to 17 are shown in formula (I), and the groups R1 to R6 are shown in Table 1:

[0090]

[0091] Table 1

[0092] Compound No. <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R3]]> <![CDATA[R4]]> <![CDATA[R5]]> <![CDATA[R6]]> 1 OH OH H H OH H 2 OH OH H <![CDATA[OCH3]]> OH H 3 H OH H OH <![CDATA[OCH3]]> H 4 H OH H <![CDATA[OCH3]]> OH H 5 H OH H H OH H 6 H OH H H <![CDATA[CF3]]> H 7 H OH H <![CDATA[OCH3]]> <![CDATA[OCH3]]> <![CDATA[OCH3]]> 8 OH <![CDATA[OCH3]]> H H OH H 9 OH <![CDATA[OCH3]]> H OH <![CDATA[OCH3]]> H 10 OH <![CDATA[OCH3]]> H <![CDATA[OCH3]]> OH H 11 OH <![CDATA[OCH3]]> H H <![CDATA[CF3]]> H 12 OH <![CDATA[CF3]]> H <![CDATA[OCH3]]> OH H 13 OH <![CDATA[CF3]]> H OH <![CDATA[OCH3]]> H 14 OH OH H OH <![CDATA[OCH3]]> H 15 H Bn H H OH H 16 H Bn H <![CDATA[OCH3]]> OH H 17 OH OH <![CDATA[OCH3]]> H H H

[0093] The structural formulas of the compounds No. 18 to 23 are shown in formula (II), and the groups R1, R2 and R7 are shown in Table 2.

[0094]

[0095] Table 2

[0096] Compound No. <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R7]]> 18 OH OH OH 19 H OH H 20 OH <![CDATA[OCH3]]> H 21 OH <![CDATA[OCH3]]> OH 22 H Bn H 23 H Bn OH

[0097] The structural formulas of the compounds No. 24 to 37 are shown in formula (III), and the groups R1 to R6 are shown in Table 3.

[0098]

[0099] Table 3

[0100]

[0101]

[0102] In Tables 1 to 3, Bn represents a benzyl group.

[0103] The structural formula of the compound No. 38 is shown in formula (38):

[0104]

[0105] The structural formula of the compound No. 39 is shown in formula (39):

[0106]

[0107] The structural formula of the compound No. 40 is shown in formula (40):

[0108]

[0109] The Chinese medicine monomer compound of isoliquiritigenin and its 39 derivatives (compounds numbered 1 to 40) were synthesized by the inventor's laboratory. The synthesis method can be found in the document Discovery of isoliquiritigenin analogues that reverse acute hepatitis by i inhibiting macrophage polarization (DOI: 10.1016 / j.bioorg.2021.105043).

[0110] Example 1

[0111] Antibacterial effects of isoliquiritigenin Chinese medicine monomer compounds and their derivatives against gonococci or drug-resistant gonococci.

[0112] (1) In vitro drug sensitivity test:

[0113] Drug-resistant gonococcal strains and culture: WHO L strain, WHO P strain and MM08 strain (international penA60.001 ceftriaxone-resistant epidemic strain) (strain drug sensitivity information see Table 8) were cultured in TM medium and GC medium recommended by CLSI and WHO at 37°C and 5% CO2 incubator. 5% skim milk was used for freeze drying, short-term preservation in -80°C refrigerator, and long-term preservation in liquid nitrogen tank.

[0114] Gonorrhea minimum inhibitory concentration (MIC) determination: The liquid dilution method was used to detect the MIC of isoliquiritigenin Chinese medicine monomer compounds and their derivatives (compounds No. 1 to 40) against different resistant gonococci. The resistant gonococci were taken out from the ultra-low temperature refrigerator, inoculated on the blood plate, and placed in a 37°C, 5% CO2 incubator for 18 to 24 hours. The gonococci were scraped, and the bacterial suspension was prepared to a concentration of 0.5 McFarland point. 150 μL of the bacterial suspension was added to 10.5 mL of liquid culture broth and mixed, and added to the drug sensitivity plate containing gradient dilutions of isoliquiritigenin Chinese medicine monomer compounds and their derivatives (100 μmol / L, 50 μmol / L, 25 μmol / L, 12.5 μmol / L, 6.25 μmol / L, 3.125 μmol / L, 1.6 μmol / L), and cultured in a 37°C, 5% CO2 incubator for 24 hours. The MIC results were observed and recorded, and blank well controls (MH broth) and growth controls (no drugs) were set at the same time. The MICs of isoliquiritigenin Chinese medicine monomer compounds and their derivatives against different drug-resistant gonococci are shown in Tables 4 to 6. Among them, 15 drugs have antibacterial effects on resistant gonococci when the concentration is ≤100μmol / L, and the results are shown in Table 4; 5 drugs have antibacterial effects on individual strains when the concentration is ≤100μmol / L, and the results are shown in Table 5; 20 drugs have no antibacterial effect on resistant gonococci when the concentration is ≤100μmol / L, and the results are shown in Table 6.

[0115] Table 4 Drugs with anti-gonococcal effects at concentrations ≤ 100 μmol / L

[0116]

[0117] Table 5 Drugs with anti-gonococcal effects on individual strains at concentrations ≤ 100 μmol / L

[0118]

[0119]

[0120] Table 6 Drugs that do not have anti-gonococcal effects when the concentration is ≤100 μmol / L

[0121]

[0122] Example 2

[0123] MIC determination of isoliquiritigenin Chinese medicine monomer compounds and their derivatives combined with different antibiotics against gonococci in vitro:

[0124] The combined drug sensitivity plate was prepared by chessboard titration method, and the working solutions of isoliquiritigenin Chinese medicine monomer compounds and their derivatives (numbered 1-40) and different antibiotics diluted in multiple ratios were added to a sterile 96-well U-shaped microplate (vertical rows are marked as A-H, horizontal rows are marked as 1-12), respectively: A-G 1-11 vertical rows are isoliquiritigenin Chinese medicine monomer compounds and their derivatives, with the highest concentration of 400μmol / L, and diluted in equal multiples in sequence; A-H 1-10 horizontal rows are penicillin (8-0.015μg / ml), ceftriaxone (1-0.002μg / ml), ciprofloxacin (8-0.015μg / ml), azithromycin (8-0.015μg / ml), cefixime (1-0.002μg / ml), tetracycline (8-0.015μg / ml) and spectinomycin (128-0.25μg / ml). The horizontal row H is for antibiotics alone, the vertical rows A to G 11 are for isoliquiritigenin Chinese medicine monomer compounds and their derivatives alone, the vertical rows A to D 12 are for growth control wells (without drugs), the vertical rows EH 12 are for culture medium control wells (MH broth), the highest concentrations of isoliquiritigenin Chinese medicine monomer compounds and their derivatives and antibiotics are the MIC values ​​of different strains, and they are diluted in multiples. After freeze-drying, seal and store in a -20°C refrigerator for later use.

[0125] Take out the drug-resistant gonococcal strain from the ultra-low temperature refrigerator, inoculate it on the blood plate, place it in a 37°C, 5% CO2 incubator for 18-24h, prepare the bacterial suspension to a concentration of 0.5 McFarland point, take 150μL of the bacterial suspension and add it to 10.5mL of liquid culture broth to mix, take 100μL and add it to the drug sensitivity plate, place it in a 37°C, 5% CO2 incubator for 24-48h, and use an enzyme reader to read the absorbance (OD) at a wavelength of 525nm. Read the OD mean of the reagent blank control wells EH 12, set cut off = (E12 + F12 + G12 + H12) OD, S / CO <1 means no growth, then the drug concentration of the well is the MIC of the drug, or observe and record the MIC manually with the naked eye. Each batch of tests is controlled by the WHO standard strain and repeated 3 times. Calculate the combined inhibition score (FIC) based on the combined drug sensitivity MIC. FIC index = MIC of isoliquiritigenin Chinese medicine monomer compound and its derivatives in combination / MIC of isoliquiritigenin Chinese medicine monomer compound and its derivatives alone + MIC of antibiotic combination / MIC of antibiotic alone. When FIC < 0.5, the two drugs have a synergistic effect; when FIC = 0.5-1, the two drugs have an additive effect; when 1 < FIC < 2, the two drugs have no effect; when FIC ≥ 2, the two drugs have an antagonistic effect.

[0126] The lowest FIC value of the combined effect of isoliquiritigenin Chinese medicine monomer compounds and their derivatives combined with different antibiotics on gonococcal strains is as follows Figure 1 As shown, the heat map shows Figure 2 shown.

[0127] Example 3

[0128] In vitro antibacterial effects on drug-resistant gonococcal clinical strains and broad-spectrum antibacterial effects:

[0129] Different types of clinically resistant strains were selected, and the MIC values ​​of isoliquiritigenin and isoliquiritigenin derivatives (No. 39) against these resistant strains were tested using the detection method of Example 1. The results are shown in Table 7.

[0130] Table 7

[0131]

[0132]

[0133] See Table 8 for strain drug sensitivity information

[0134] Table 8 Antimicrobial susceptibility information of strains

[0135]

[0136] PPNG: Penicillinase-producing Neisseria gonorrhoeae. / : indicates not tested.

[0137] The drug sensitivity interpretation standard was interpreted with reference to the WHO resistance breakpoint standard. For penicillin, tetracycline and ciprofloxacin, MIC ≥ 1 mg / L was resistant and < 1 mg / L was sensitive. For ceftriaxone, MIC ≥ 0.125 mg / L was resistant and < 0.125 mg / L was sensitive. For cefixime, MIC ≥ 0.25 mg / L was resistant and < 0.25 mg / L was sensitive. For azithromycin, MIC ≥ 2 mg / L was resistant and < 2 mg / L was sensitive.

[0138] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of isoliquiritigenin and its derivatives in the preparation of anti-gonococcal drugs; The isoliquiritigenin and its derivatives include compound 1, compound 2, compound 4, compound 5, compound 6, compound 8, compound 9, compound 10, compound 11, compound 13, compound 14, compound 15, compound 117, compound 20, compound 21, compound 22, compound 24, compound 37, compound 39 and compound 40; Wherein compound 1 is isoliquiritigenin, and its structural formula is shown in formula (1); The structural formulas of Compound 2, Compound 4, Compound 5, Compound 6, Compound 8, Compound 9, Compound 110, Compound 11, Compound 13, Compound 14, Compound 15 and Compound 17 are shown in Formula (I): Where R1, R2, R3, R4, R5 and R6 are shown in the following table: The structural formulas of Compound 20, Compound 21 and Compound 22 are shown in Formula (II): Where R1, R2 and R3 are shown in the following table: The structural formulas of compound 24 and compound 37 are shown in formula (III): Where R1, R2, R3, R4, R5 and R6 are shown in the following table: The structural formula of compound 39 is shown in formula (39): The structural formula of compound 40 is shown in formula (40): Compound 40:

2. The use according to claim 1, characterized in that: The application includes applying at least one of the following isoliquiritigenin and its derivatives to prepare anti-gonococcal drugs: compound 1, compound 2, compound 5, compound 8, compound 9, compound 10, compound 13, compound 14, compound 15, compound 20, compound 21, compound 22, compound 37, compound 39 and compound 40; Optionally, the effective concentrations of Compound 1, Compound 2, Compound 5, Compound 8, Compound 9, Compound 10, Compound 13, Compound 14, Compound 15, Compound 20, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40 in the anti-gonococcal drug are independently at least 10 μmol / L, or at least 25 μmol / L, or at least 50 μmol / L, or at least 100 μmol / L.

3. The use according to claim 2, characterized in that: The application includes applying at least one of the following isoliquiritigenin derivatives to prepare an anti-gonococcal drug: compound 1, compound 9, compound 10, compound 15, compound 21, compound 22, compound 37, compound 39 and compound 40; Optionally, the effective concentrations of Compound 1, Compound 9, Compound 10, Compound 15, Compound 21, Compound 22, Compound 37, Compound 39 and Compound 40 in the anti-gonococcal drug are independently at least 10 μmol / L, or at least 25 μmol / L, or at least 50 μmol / L.

4. The use according to any one of claims 1 to 3, characterized in that: The gonococci include drug-resistant gonococci; Optionally, the drug-resistant gonococci include gonococci resistant to at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin and ceftriaxone.

5. The use according to claim 4, characterized in that: The application includes applying at least one of the following isoliquiritigenin and its derivatives to prepare an anti-drug-resistant gonococcal drug: compound 1, compound 5, compound 6, compound 9, compound 10, compound 15, compound 21, compound 37, compound 39 and compound 40; Optionally, the anti-resistant gonococcal drug further comprises an antibiotic; Optionally, the antibiotics include at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin, ceftriaxone and cefixime.

6. The use according to claim 5, characterized in that: The application includes applying penicillin and at least one of the following isoliquiritigenin derivatives to prepare drugs against drug-resistant gonococci: compound 6, compound 10, compound 21 and compound 39; Or, the application includes applying tetracycline and at least one of the following isoliquiritigenin derivatives to prepare an anti-drug-resistant gonococcal drug: compound 6, compound 10, compound 21 and compound 39; Or, the application includes applying ciprofloxacin and at least one of the following isoliquiritigenin derivatives to prepare an anti-drug-resistant gonococcal drug: compound 6, compound 21 and compound 39; Or, the application includes applying azithromycin and at least one of the following isoliquiritigenin derivatives to prepare an anti-drug-resistant gonococcal drug: compound 6, compound 10, compound 15, compound 21 and compound 39; Or, the application includes applying ceftriaxone and at least one of the following isoliquiritigenin derivatives to prepare an anti-drug-resistant gonococcal drug: compound 6, compound 10, compound 21 and compound 39; Or, the application includes applying cefixime and compound 6 to prepare drugs against drug-resistant gonococci.

7. An antigonococcal drug, characterized in that: Contains (A) and (B): (A) Antibiotics: (B) one or more of isoliquiritigenin and its derivatives as described in any one of claims 1 to 6.

8. The antigonococcal drug according to claim 7, characterized in that The antibiotics include at least one of penicillin, tetracycline, ciprofloxacin, spectinomycin, azithromycin, ceftriaxone and cefixime.

9. The antigonococcal drug according to claim 7, characterized in that The gonococcal drug comprises at least one of the following isoliquiritigenin and its derivatives: compound 1, compound 5, compound 6, compound 9, compound 10, compound 15, compound 21, compound 37, compound 39 and compound 40.

10. The antigonococcal drug according to claim 8 or 9, characterized in that The antigonococcal drug comprises penicillin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39; Or, the antigonococcal drug comprises tetracycline and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39; Or, the antigonococcal drug comprises ciprofloxacin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 21 and Compound 39; Or, the antigonococcal drug comprises azithromycin and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 15, Compound 21 and Compound 39; Or, the antigonococcal drug comprises ceftriaxone and at least one of the following isoliquiritigenin derivatives: Compound 6, Compound 10, Compound 21 and Compound 39; Alternatively, the antigonococcal drug comprises cefixime and compound 6.

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