Amphiphilic resorcinol calixarene based on guanidino carbonyl pyrrole functionalization and nanoparticles containing amphiphilic resorcinol calixarene

Abstract

The invention provides amphiphilic resorcinol calixarene based on guanidino carbonyl pyrrole functionalization and a nano particle containing the amphiphilic resorcinol calixarene. Firstly, nanoparticles are constructed on the basis of amphiphilic resorcinol calixarene, then antibiotic norfloxacin is entrapped in the nanoparticles, and binary nanoparticles with a synergistic antibacterial effect are prepared and are used for efficient antibiosis. The nanoparticles show excellent synergistic antibacterial effect and biocompatibility under the condition that the concentration is far lower than the minimum inhibitory concentration of subjects and guests, and the problem of drug resistance of bacteria is avoided to the greatest extent due to the use of low-dose antibiotics.

Description

Technical Field

[0001] This invention relates to the field of supramolecular materials chemistry, specifically to a guanidine carbonyl pyrrole-functionalized amphiphilic resorcinol calixarene and nanoparticles containing it. Background Technology

[0002] Bacterial infections have always been a serious threat to human public health. For a time after their invention, antibiotics were effective at killing bacteria, protecting humans from infection. However, due to the misuse and overuse of antibiotics, bacteria have evolved and developed resistance, significantly reducing the effectiveness of traditional antibiotic treatments. The emergence of drug-resistant bacteria has forced researchers to seek new strategies to combat microbial infections.

[0003] Supramolecular systems are widely used as a novel antibacterial strategy due to their simple preparation and advantages such as dynamism, reversibility, and precise controllability. Supramolecular macrocycles can encapsulate guest molecules or self-assemble into nanoparticles, offering unique advantages in the delivery and protection of antibacterial agents; therefore, antibacterial systems based on supramolecular macrocycles have been extensively reported. Guanidinocarbonylpyrrole (GCP), containing arginine residues, can be used as an arginine substitute in biological fields, such as protein recognition and gene transfection. In this process, GCP exhibits excellent membrane binding ability and biocompatibility, showing the potential to bind to and disrupt bacterial membranes. The orthogonal combination of supramolecular macrocycles and GCP can significantly enhance the biocompatibility of macrocyclic compounds and endow them with antibacterial properties, providing a new solution to the problems of antibiotic abuse and bacterial resistance. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the purpose of this invention is to provide a method for preparing binary nanoparticles based on guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene and their application.

[0005] The technical solution provided by this invention is as follows: In a first aspect, the present invention provides a guanidinocarbonylpyrrole compound, the structural formula of which is as follows: .

[0006] Secondly, the present invention provides a method for synthesizing the guanidinocarbonylpyrrole compound as described above, comprising the following steps: (a) Pyrrole reacts with a carboxylic acid derivative to form compound A; the structural formula of compound A is as follows: ; (b) dissolving compound A in a halogenated hydrocarbon solvent to obtain a compound A solution under inert gas protection; adding a solution of an alkoxyl alkali metal compound into the compound A solution dropwise to react, to form compound B; the structural formula of the compound B is as follows: ; (c) performing formylation reaction on the compound B in the presence of a Vilsmeier-Haack reagent, to form compound C; the structural formula of the compound C is as follows: ; (d) oxidizing the compound C in the presence of an oxidant, to form a carboxylic acid compound D; the structural formula of the compound D is as follows: ; (e) reacting a guanidine or a guanidine salt with a protection reagent under alkaline conditions, to form a protected guanidine compound E; the structural formula of the compound E is as follows: ; (f) performing condensation reaction on the carboxylic acid compound D and the protected guanidine compound E in the presence of a coupling agent and an organic base, to form compound F; the structural formula of the compound F is as follows: ; (g) performing catalytic hydrogenation deprotection reaction on the compound F in the presence of a catalyst and a base in a reducing atmosphere, to form compound G; the structural formula of the compound G is as follows: ; (h) performing final acidification reaction on the compound G under acidic conditions, to obtain the guanidyl carbonyl pyrrole compound.

[0007] In step (a), the carboxylic acid derivative is trichloroacetyl chloride; and / or, the molar ratio of the pyrrole to the carboxylic acid derivative is 1:1.1-1:1.5.

[0008] In step (b), the reaction conditions of the solution of the alkoxyl alkali metal compound with the compound A solution are: 25-35 ℃, and the reaction time is 1-5 hours; wherein, the molar ratio of the compound A to the alkali metal is 1:0.1-1:0.3.

[0009] The preparation method of the solution of the alkoxyl alkali metal compound is: stirring the alkali metal in aralkyl alcohol at 40-50 ℃, to generate the solution of the alkoxyl alkali metal compound by the reaction of the aralkyl alcohol with the alkali metal. The aralkyl alcohol is benzyl alcohol; and the alkali metal is sodium.

[0010] In step (c), the Vilsmeier-Haack reagent is generated by the reaction of N , Ndimethylformamide.

[0011] The molar ratio of the compound B to phosphorus oxychloride is 1:1.5-1:2.5.

[0012] In step (d), the oxidant is potassium permanganate; and / or, the molar ratio of the compound C to the oxidant is 1:1-1:3.

[0013] In step (e), the protecting agent is di-tert-butyl dicarbonate; and / or, the guanidine salt is guanidine hydrochloride; and / or, the molar ratio of the protecting agent to guanidine or guanidine salt is 1:4-1:7.

[0014] In step (f), the coupling agent is 6-chlorobenzotriazole-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU), and the molar ratio of the compound D to HCTU is 1:1-1.5; and / or, the organic base is N methylmorpholine, and the molar ratio of the compound D to N methylmorpholine is 1:2-3; and / or, the molar ratio of the compound D to the compound E is 1:2-2.5.

[0015] In step (g), the catalyst is palladium on carbon; and / or, the base is triethylamine, and the molar ratio of the compound F to triethylamine is 1:1-2; and / or, the reducing atmosphere is hydrogen atmosphere.

[0016] As an embodiment of the present application, step (a) is specifically that, under the protection of inert gas, trichloroacetyl chloride is dissolved in an ether solvent (such as anhydrous diethyl ether), and pyrrole is added dropwise, and the reaction is carried out at 25-35 ℃ for 12-24 hours to obtain the compound A; the molar ratio of pyrrole to trichloroacetyl chloride is 1:1.1-1:1.5.

[0017] As an embodiment of the present application, step (b) is specifically that, under the protection of inert gas, the compound A is dissolved in a halogenated hydrocarbon solvent (chloroform) to prepare a compound A solution; sodium is stirred in benzyl alcohol at 40-50 ℃ to prepare a sodium benzyloxide solution, which is added dropwise into the compound A solution, and the reaction is carried out at 25-35 ℃ for 1-5 hours to obtain the compound B; the molar ratio of the compound A to sodium is 1:0.1-1:0.3.

[0018] As an embodiment of the present application, step (c) is specifically that, under the protection of inert gas, the compound B is dissolved in dichloromethane; phosphorus oxychloride is added dropwise into N N ​- stirring for 20-50 minutes in dimethylformamide to obtain Vilsmeier reagent; drop the Vilsmeier reagent into a dichloromethane solution containing compound B, and react for 10-24 hours at 25-35 ℃ to obtain compound C; the molar ratio of compound B to phosphorus oxychloride is 1:1.5-1:2.5.

[0019] As an embodiment of the present application, step (d) is specifically dissolving compound C in acetone to form a reaction solution under inert gas protection; then, drop a potassium permanganate mixed solution of acetone / water into the reaction solution, and react for 12 hours at 30-50 ℃, and then continue to react for 1-2 hours at 25-35 ℃, and obtain carboxylic acid compound D after post-treatment. The molar ratio of compound C to potassium permanganate is 1:1-1:3.

[0020] As an embodiment of the present application, step (e) is specifically dissolving guanidine hydrochloride in sodium hydroxide solution under inert gas protection, and then drop di-tert-butyl dicarbonate acetonitrile solution into the solution, and react for 10-24 hours at 25-35 ℃ to obtain guanidine compound E; the molar ratio of di-tert-butyl dicarbonate to guanidine hydrochloride is 1:4-1:7.

[0021] As an embodiment of the present application, step (f) is specifically dissolving compound D, 6-chlorobenzotriazole-1,1,3,3-tetramethyluron hexafluorophosphate and N - methylmorpholine in N , N - dimethylformamide, stirring for 30-40 minutes, adding guanidine compound E, and reacting for 10-24 hours at 25-35 ℃ to obtain compound F; the molar ratio of compound D to 6-chlorobenzotriazole-1,1,3,3-tetramethyluron hexafluorophosphate is 1:1-1:1.5, the molar ratio of compound D to methylmorpholine is 1:2-1:3, and the molar ratio of compound D to guanidine compound E is 1:2-1:2.5. N - dimethylformamide, stirring for 30-40 minutes, adding guanidine compound E, and reacting for 10-24 hours at 25-35 ℃ to obtain compound F; the molar ratio of compound D to 6-chlorobenzotriazole-1,1,3,3-tetramethyluron hexafluorophosphate is 1:1-1:1.5, the molar ratio of compound D to methylmorpholine is 1:2-1:3, and the molar ratio of compound D to guanidine compound E is 1:2-1:2.5.

[0022] As an embodiment of the present application, step (g) is specifically dissolving compound F and palladium-carbon in methanol under hydrogen protection, adding triethylamine, and reacting for 10-24 hours at 30-50 ℃ to obtain compound G; the molar ratio of compound F to triethylamine is 1:1-1:2.

[0023] As an embodiment of the present application, step (h) is specifically dissolving compound G in deionized water, adding dilute hydrochloric acid, and reacting for 10-30 minutes at 25-35 ℃ to obtain compound H, i.e. the guanidyl carbonyl pyrrole compound.

[0024] In the present application, the synthetic route of guanidyl carbonyl pyrrole compound is as follows: .

[0025] As an embodiment of the present application, the specific synthesis steps of guanidyl carbonyl pyrrole compound are as follows: a. Dissolve trichloroacetyl chloride in anhydrous ether under inert gas protection, drop pyrrole, and react at 25-35 ℃ for 12-24 hours to obtain compound A; b. Dissolve compound A in chloroform under inert gas protection, prepare sodium benzyloxide solution by placing sodium in benzylic alcohol, drop the above solution, and react at 25-35 ℃ for 1-5 hours to obtain compound B; c. Dissolve compound B in dichloromethane under inert gas protection, drop trichloro phosphine into N , N dimethylformamide, stir for 20-50 minutes, drop the above solution, and react at 25-35 ℃ for 10-24 hours to obtain compound C; d. Dissolve compound C in acetone under inert gas protection, drop potassium permanganate acetone / water solution, react at 30-50 ℃ for 1-2 hours, and react at 25-35 ℃ for 1-2 hours to obtain compound D; e. Dissolve guanidine hydrochloride in sodium hydroxide solution under inert gas protection, drop di-tert-butyl dicarbonate acetonitrile solution, and react at 25-35 ℃ for 10-24 hours to obtain compound E; f. Dissolve compound D, 6-chlorobenzotriazole-1,1,3,3-tetramethyluron hexafluorophosphate and N methylmorpholine in N , N dimethylformamide, stir for 30-40 minutes, add compound E, and react at 25-35 ℃ for 10-24 hours to obtain compound F; g. Dissolve compound F and palladium carbon in methanol under hydrogen protection, add triethylamine, and react at 30-50 ℃ for 10-24 hours to obtain compound G; h. Dissolve compound G in deionized water, add dilute hydrochloric acid, and react at 25-35 ℃ for 10-30 minutes to obtain compound H.

[0026] In a third aspect, the present application provides a guanidyl carbonyl pyrrole functionalized amphiphilic resorcinol calixarene, whose structural formula is shown in formula (1):

[0027] formula (1); wherein the number of carbon atoms n of the alkyl chain is 1-10.

[0028] As an embodiment of the present application, the structure of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinol calixarene is shown as follows:

[0029] In a fourth aspect, the present application provides a preparation method of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinol calixarene as described above, comprising the following steps: S1, reacting a polyphenol compound with a cyclization reagent under acidic conditions to form a cyclic compound I, the structure of the compound I is shown as formula (2):

[0030] Formula (2); S2, reacting the cyclic compound I with a halomethylating reagent under basic conditions to obtain a compound J, the structure of the compound J is shown as formula (3):

[0031] Formula (3); S3, performing a halogen exchange reaction on the compound J in the presence of a brominating reagent to obtain a bromomethyl compound K; the structure of the compound K is shown as formula (4):

[0032] Formula (4); S4, reacting the bromomethyl compound K with an amine source to form an amine-based compound L; the structure of the compound L is shown as formula (5):

[0033] Formula (5); S5, performing a condensation reaction on the amine-based compound L with a guanidylcarbonyl pyrrole compound (compound H) in the presence of a coupling agent and an organic base, and removing the protecting group in the presence of trifluoroacetic acid to obtain the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinol calixarene, denoted as GCP-CA.

[0034] In step S1, the polyphenol compound is resorcinol; and / or, the cyclization reagent is 2,3-dihydrofuran. The molar ratio of the resorcinol to 2,3-dihydrofuran is 1:0.9~1.1; the reaction is carried out at a temperature range of -20 ℃ to 50 ℃, the acidic conditions are provided by concentrated hydrochloric acid; the reaction time is 12 hours to 5 days.

[0035] In step S2, the basic condition is provided by potassium carbonate, the molar ratio of compound I to potassium carbonate is 1:9-11; and / or, the halomethylating agent is bromochloromethane; the molar ratio of compound I to bromochloromethane is 1:19-21; the reaction temperature is 45-55℃, and the reaction time is 4-6 days.

[0036] In step S3, the brominating agent is a combination of triphenylphosphine and carbon tetrabromide; the molar ratio of compound J to triphenylphosphine is 1:5-7; the molar ratio of compound J to carbon tetrabromide is 1:5-7; the reaction is carried out at 20-30℃, and the reaction time is 10-24 hours.

[0037] Step S4 includes the following steps: S4a, reacting compound K with a phthalimide salt to obtain a protected amine group intermediate; the molar ratio of compound K to the phthalimide salt is 1:5-7; the reaction temperature of this step is 35-45℃, and the reaction time is 20-28 hours; S4b, deprotecting the protected amine group intermediate using a hydrazine reagent to obtain the amine compound L; the deprotection reaction is carried out at 70-90℃, and the reaction time is 10-24 hours.

[0038] In S4a, the phthalimide salt is potassium phthalimide; and / or, the hydrazine reagent is hydrazine hydrate.

[0039] Step S5 includes the following steps: S5a, activating the carboxyl group of the guanidinyl carbonyl pyrrole compound in the presence of a coupling agent and an organic base; the molar ratio of the carboxyl group of the guanidinyl carbonyl pyrrole compound, the coupling agent and the organic base is 1:1.0-1.2:1.2-1.5, and the activation time is 30-60 minutes; S5b, condensing the activated carboxyl group of the guanidinyl carbonyl pyrrole compound with the amine compound L to obtain a protected calixarene derivative; the molar ratio of the carboxyl group of the guanidinyl carbonyl pyrrole compound to the amine compound L is 3.5-4.5:1; the condensation reaction is carried out at 20-30℃, and the reaction time is 20-28 hours; S5c, removing the protecting group in the presence of trifluoroacetic acid to obtain the final product guanidinyl carbonyl pyrrole functionalized amphiphilic resorcinol calixarene (GCP-CA); the deprotection reaction is carried out at 20-30℃, and the reaction time is 1-3 hours.

[0040] In step S5a, the coupling agent is 6-chlorobenzotriazole-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU); and / or, the organic base is N ,N - Diisopropylethylamine (DIPEA).

[0041] wherein CF3COO - in formula (1) is derived from the deprotection reaction in step S5c. In this step, trifluoroacetic acid (CF3COOH) is used to remove the protecting group under acidic conditions. After the reaction, the trifluoroacetic acid in the system exists in the form of CF3COO - , thereby introducing the CF3COO - group.

[0042] As an embodiment of the present application, the preparation method of the guanidyl carbonyl pyrrole functionalized amphiphilic catechol calixarene (GCP-CA) comprises the following steps: (1) Under the protection of inert gas, resorcinol (1.0 equiv.) is dissolved in a mixed solution of methanol and concentrated hydrochloric acid, 2,3-dihydrofuran (1.0 equiv.) is slowly added dropwise, and the reaction is carried out at 30-80 ℃ for 2-5 days. After the reaction is completed, the compound I is obtained by post-treatment; (2) Under the protection of inert gas, compound I (1.0 equiv.) is dissolved in N , N dimethylacetamide, bromochloromethane (20 equiv.) and potassium carbonate (10 equiv.) are added, and the reaction is carried out at 30-80 ℃ for 2-5 days. After the reaction is completed, the compound J is obtained by post-treatment; (3) Under the protection of inert gas, compound J (1.0 equiv.) is dissolved in dichloromethane, triphenylphosphine (6 equiv.) and carbon tetrabromide (6 equiv.) are added in sequence, and the reaction is carried out at 25-40 ℃ for 10-24 hours. After the reaction is completed, the compound K is obtained by post-treatment; (4) Under the protection of inert gas, compound K (1.0 equiv.) and potassium phthalimide (6 equiv.) are dissolved in N , N dimethylformamide, and the reaction is carried out at 30-60 ℃ for 1-2 days. After the reaction is completed, the solvent is removed, and the obtained solid is dissolved in ethanol under the protection of inert gas, hydrazine hydrate is added, and the reaction is carried out at 60-100 ℃ for 10-24 hours. After the reaction is completed, the compound L is obtained by post-treatment; (5) Under the protection of inert gas, guanidyl carbonyl pyrrole compound (compound H, 4.0 equiv.), 6-chlorobenzotriazole-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU, 4.5 equiv.) and N , N diisopropylethylamine (DIPEA, 5.0 equiv.) are dissolved in N, N In dimethylformamide, stir for 30-40 minutes to complete activation; then add compound L (1.0 equiv.) and react at 25-35 °C for 1-2 days; after the reaction is complete, remove the solvent, dissolve the obtained solid in dichloromethane under inert gas protection, add trifluoroacetic acid, and react at 25-35 °C for 1-4 hours. After the reaction is complete, post-treatment yields the guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene compound GCP-CA.

[0043] The synthetic route for guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene host molecules is as follows:

[0044] As one embodiment of the present invention, the specific synthetic steps of the guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene host are as follows: S1i. Under inert gas protection, resorcinol is dissolved in a mixed solution of methanol and concentrated hydrochloric acid, and 2,3-dihydrofuran is added dropwise. The mixture is reacted at 30-80 °C for 2-5 days to obtain compound I. S1j, under inert gas protection, dissolve compound I in... N , N -Dimethylacetamide, with the addition of bromochloromethane, reacted at 30-80℃ for 2-5 days to give compound J; S1k, under inert gas protection, compound J is dissolved in dichloromethane, and triphenylphosphine and carbon tetrabromide are added sequentially. The reaction is carried out at 25-40 °C for 10-24 hours to obtain compound K; S1l, under inert gas protection, dissolve compound K and potassium phthalimide in... N , N -Dimethylformamide, reacted at 30-60 °C for 1-2 days, the solid obtained after solvent removal was dissolved in ethanol under inert gas protection, hydrazine hydrate was added, and reacted at 60-100 °C for 10-24 hours to obtain compound L; S1m, under inert gas protection, compound H, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate and N , N -Diisopropylethylamine dissolves in N , N Dimethylformamide was stirred for 30-40 minutes, compound L was added, and the mixture was reacted at 25-35 °C for 1-2 days. The solid obtained after solvent removal was dissolved in dichloromethane under inert gas protection, and trifluoroacetic acid was added. The mixture was reacted at 25-35 °C for 1-4 hours to obtain compound GCP-CA.

[0045] Preferably, in step S1i, the molar ratio of resorcinol to 2,3-dihydrofuran is 1:1 to 1:1.5; in step S1j, the molar ratio of compound I to potassium carbonate is 1:10 to 1:15, and the molar ratio of compound I to bromochloromethane is 1:20 to 1:30; in step S1k, the molar ratio of compound J to triphenylphosphine is 1:6 to 1:8, and the molar ratio of compound J to carbon tetrabromide is 1:6 to 1:8; in step S1l, the molar ratio of compound K to potassium phthalimide is 1:6 to 1:8; in step S1m, the molar ratio of compound L to 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate is 1:5 to 1:7; and the molar ratio of compound L to... N , N The molar ratio of diisopropylethylamine is 1:5 to 1:7; the molar ratio of compound L to compound H is 1:4 to 1:6.

[0046] Furthermore, this invention provides a method for preparing binary nanoparticles based on guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene, comprising the following steps: Step 1: Dissolve guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene GCP-CA and norfloxacin in organic solvents to prepare GCP-CA mother liquor and norfloxacin mother liquor, respectively. Step 2: Mix the GCP-CA mother liquor with norfloxacin mother liquor to obtain a host-guest mixture; Step 3: Inject the host-guest mixture into an aqueous medium and mix evenly to obtain the binary nanoparticle solution based on guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene.

[0047] The organic solvent is dimethyl sulfoxide. N , N - One or more of dimethylformamide and ethanol; preferably, the organic solvent is dimethyl sulfoxide.

[0048] The aqueous medium is deionized water or a buffer salt solution.

[0049] In step 1, the concentration of the GCP-CA mother liquor is 10-25 mM; the concentration of the norfloxacin mother liquor is 50-150 µM.

[0050] In step 2, the volume ratio of the GCP-CA mother liquor to the norfloxacin mother liquor is (1.5-2.5):1.

[0051] In step 3, the volume ratio of the host-guest mixture to the aqueous medium is 1:(45-55).

[0052] In the fifth aspect, the application further provides a use of the binary nanoparticle based on guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene in the preparation of an antibacterial drug.

[0053] The antibacterial drug is used for inhibiting Staphylococcus aureus and Escherichia coli.

[0054] In the solution of the binary nanoparticle used for inhibiting Staphylococcus aureus, the concentration of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene is 0.2-0.3 mM, and the concentration of norfloxacin is 0.5-0.7 µM.

[0055] In the solution of the binary nanoparticle used for inhibiting Escherichia coli, the concentration of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene is 0.1-0.15 mM, and the concentration of norfloxacin is 0.2-0.4 µM.

[0056] The structure of the antibiotic norfloxacin is as follows: .

[0057] The binary nanoparticle based on guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene provided by the application comprises guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene and antibiotic norfloxacin. The norfloxacin can be loaded into the nanoparticle formed by self-assembly of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene to form a binary nanoparticle and achieve high-efficiency synergistic antibacterial effect.

[0058] The lower edge of the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene is covalently connected with four positively charged GCP groups, and the guanidylcarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene can self-assemble into a nanoparticle under the driving of hydrophobic interaction. The nanoparticle can be combined with the surface of a bacterium with negative charge through electrostatic interaction, and damage the bacterium by affecting the function of the bacterial membrane. In addition, the host can further form a binary nano-assembly with a positive charge on the outer layer by interacting with the guest molecules through hydrogen bonds and the like. After the host destroys the bacterial membrane, the guest norfloxacin enters the inside of the bacterium to inhibit the synthesis and replication of bacterial DNA. After the synergistic action of the two, high-efficiency synergistic antibacterial performance can be achieved at much lower than the minimum inhibitory concentration of each. In addition, the introduction of the GCP group can significantly reduce the cytotoxicity of the resorcinarene calixarene, so that the resorcinarene calixarene has good biocompatibility.

[0059] Compared with the prior art, the application has the following beneficial effects: (1) The application adopts a covalent connection mode to construct a novel amphiphilic resorcinol calixarene, the introduction of GCP endows the resorcinol calixarene with antibacterial ability and biocompatibility; (2) The synergistic effect greatly reduces the dosage of norfloxacin, and the treatment effect is good, the side effect is low, and the problem of drug resistance is avoided; (3) The method adopts a non-covalent bond assembly mode to encapsulate and deliver the therapeutic drug, avoids the interference to the activity of the drug, and improves the stability. BRIEF DESCRIPTION OF DRAWINGS

[0060] Other features, objects and advantages of the application will become more apparent from the following detailed description of non-limiting embodiments, made with reference to the following drawings: Figure 1 The H NMR spectrum of compound A is as follows: 1 The H NMR spectrum of compound A is as follows: Figure 2 The H NMR spectrum of compound B is as follows: 1 The H NMR spectrum of compound B is as follows: Figure 3 The H NMR spectrum of compound C is as follows: 1 The H NMR spectrum of compound C is as follows: Figure 4 The H NMR spectrum of compound D is as follows: 1 The H NMR spectrum of compound D is as follows: Figure 5 The H NMR spectrum of compound E is as follows: 1 The H NMR spectrum of compound E is as follows: Figure 6 The H NMR spectrum of compound F is as follows: 1 The H NMR spectrum of compound F is as follows: Figure 7 The H NMR spectrum of compound G is as follows: 1 The H NMR spectrum of compound G is as follows: Figure 8 The H NMR spectrum of compound H is as follows: 1 The H NMR spectrum of compound H is as follows: Figure 9 The H NMR spectrum of compound I is as follows: 1 The H NMR spectrum of compound I is as follows: Figure 10 The H NMR spectrum of compound J is as follows: 1 The H NMR spectrum of compound J is as follows: Figure 11 The H NMR spectrum of compound K is as follows: 1 The H NMR spectrum of compound K is as follows: Figure 12 The C NMR spectrum of compound K is as follows: 13 The C NMR spectrum of compound K is as follows: Figure 13 The HR-ESI-MS spectrum of compound K is as follows: Figure 14 The H NMR spectrum of compound L is as follows: 1 The H NMR spectrum of compound L is as follows: Figure 15 HR-ESI-MS spectrum of compound L; 13 H NMR spectrum; Figure 16 HR-ESI-MS spectrum of compound L; Figure 17 H NMR spectrum of compound GCP-CA; 1 H NMR spectrum; Figure 18 HR-ESI-MS spectrum of compound GCP-CA; 13 H NMR spectrum; Figure 19 HR-ESI-MS spectrum of compound GCP-CA; Figure 20 SEM image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene nanoparticle; Figure 21 DLS image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene nanoparticle; Figure 22 SEM image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticle; Figure 23 DLS image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticle; Figure 24 Bacteriostatic colony image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticle; Figure 25 Bacteriostatic effect image of guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticle; Figure 26 SEM image of bacteria after co-incubation with guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticle. DETAILED DESCRIPTION

[0061] The application will be described in greater detail below with reference to specific embodiments. The following examples are helpful for those skilled in the art to further understand the application, but do not limit the application in any form. It should be noted that those skilled in the art can make several changes and improvements without departing from the concept of the application. These all belong to the protection scope of the application.

[0062] The application discloses a binary nanoparticle based on guanidylcarbonylpyrrole functionalized amphiphilic resorcinol calixarene The preparation method comprises the following steps: firstly, a guanidyl carbonyl pyrrole functionalized amphiphilic resorcinol calixarene host molecule is designed and synthesized; then, norfloxacin is selected as a guest molecule, and a binary nanoparticle is assembled through host-guest interaction. The biological experiment result shows that the nanoparticle can kill bacteria under the condition of far lower than the minimum inhibitory concentration of the host-guest, has excellent synergistic antibacterial activity and biocompatibility. The method provides a novel antibacterial strategy, and has important significance for preventing the emergence of drug-resistant bacteria.

[0063] Example 1 Preparation of guanidinocarbonylpyrrole functionalized amphiphilic resorcinarene calixarene host molecules 1. The synthesis process of a guanidyl carbonyl pyrrole compound is as follows:

[0064] (1) Synthesis of compound A: trichloroacetyl chloride (1.1 equiv.) is dissolved in diethyl ether. Prolonged dropwise addition of pyrrole (1 equiv.) is carried out at room temperature for 1 hour. Potassium carbonate solution is added, and liquid separation is carried out. Anhydrous sodium sulfate and activated carbon are dried and decolorized. Filtration is carried out, and the solvent is removed by distillation under reduced pressure. Hexane recrystallization is carried out to obtain the product A in the form of a gray-white solid. 1 H NMR (400 MHz, CDCl3, 298 K) δ 9.43 (s, 1H), 7.41 – 7.37 (m, 1H), 7.17 (dd, J = 3.4, 1.6 Hz, 1H), 6.41 –6.37 (m, 1H). The H NMR spectrum of compound A is as shown in 1 H NMR (400 MHz, CDCl3, 298 K) δ 9.43 (s, 1H), 7.41 – 7.37 (m, 1H), 7.17 (dd, Figure 1 .

[0065] (2) Synthesis of compound B: sodium (0.2 equiv.) is stirred in benzyl alcohol at 45 DEG C until complete reaction. Compound A (1 equiv.) is dissolved in chloroform under a nitrogen atmosphere. The prepared sodium benzyl alcohol solution is added, and reaction is carried out at room temperature for 1 hour. Dilute hydrochloric acid (1 N) is added, and stirring is carried out for 10 min. Liquid separation is carried out, and deionized water is washed. Anhydrous sodium sulfate is dried. Filtration is carried out, and the solvent is removed by distillation under reduced pressure. The obtained crude product is separated and purified by column chromatography to obtain the product B in the form of a white solid. 1 H NMR (400 MHz, CDCl3, 298 K) δ 9.43 (s, 1H), 7.41 – 7.37 (m, 1H), 7.17 (dd, J = 3.4, 1.6 Hz, 1H), 6.41 –6.37 (m, 1H). The H NMR spectrum of compound A is as shown in 1 H NMR (400 MHz, CDCl3, 298 K) δ 9.43 (s, 1H), 7.41 – 7.37 (m, 1H), 7.17 (dd, Figure 2 .

[0066] (2) Synthesis of compound C: Vilsmeier reagent was prepared by slowly adding phosphine oxychloride (2 equiv.) dropwise into DMF and stirring for 30 min. Compound B (1 equiv.) was dissolved in dichloromethane under a nitrogen atmosphere. The system temperature was lowered to an ice-salt bath. At 20 °C, the Vilsmeier reagent was slowly added dropwise (approximately 20 min), and the mixture was stirred for 3 hours. The ice-salt bath was removed, and the reaction was allowed to proceed overnight at room temperature. Saturated sodium bicarbonate solution was added dropwise, and the mixture was refluxed for 15 min. The mixture was filtered, separated, and the organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation, and the crude product was purified by column chromatography to obtain a gray solid product C. 1 H NMR (400 MHz, CDCl3, 298 K) δ 9.86 (s, 1H), 9.66 (s, 1H), 7.45 – 7.35 (m, 5H), 6.98 (dd, J = 3.9, 2.5 Hz, 1H), 6.95 – 6.93 (m, 1H), 5.35 (s, 2H). Compound C's... 1 HNMR spectrum as follows Figure 3 .

[0067] (3) Synthesis of compound D: Compound C (1 equiv.) was dissolved in acetone under a nitrogen atmosphere. Potassium permanganate (2 equiv.) was dissolved in an acetone / water mixture (1:1). v / v The solution was slowly added dropwise to the above solution. The reaction was carried out at 40 °C for 1 hour, followed by a reaction at room temperature for 1 hour. The mixture was then filtered and washed with sodium hydroxide solution (5%). The resulting filtrate was acidified with dilute hydrochloric acid (5%) to pH = 4, filtered, washed with deionized water, and dried under vacuum at 80 °C overnight to obtain a white solid product D. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ12.59 (s, 1H), 7.45 (d, J = 7.1 Hz, 2H), 7.35 (m, 3H), 6.83 (dd, J = 3.7, 2.4 Hz, 1H), 6.75 (dd, J = 3.7, 2.4 Hz, 1H), 5.28 (s, 2H). Compound D's 1 H NMR spectrum as shown Figure 4 .

[0068] (4) Synthesis of compound E: Guanidine hydrochloride (5 equiv.) was dissolved in sodium hydroxide solution (6 M) under a nitrogen atmosphere. Di-tert-butyl dicarbonate (1 equiv.) was dissolved in acetonitrile and slowly added dropwise to the above solution. The reaction was carried out at room temperature for 20 hours. The solvent was removed by vacuum distillation. Ethyl acetate was added to the resulting solid, which was washed with deionized water, washed with saturated sodium chloride solution, and dried over anhydrous sodium sulfate. The solid was filtered and the solvent was removed by vacuum distillation to give a white solid product E. 1 H NMR (400 MHz, DMSO- d δ 6.79 (s, 4H), 1.33 (s, 9H) at K 6, 298 K. Compound E... 1 H NMR spectrum as shown Figure 5 .

[0069] (5) Synthesis of compound F: Compound D (1 equiv.), 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate (1.1 equiv.), and N 2-Methylmorpholine (2.2 equiv.) was dissolved in DMF and stirred for 30 min. Compound E (2 equiv.) was added, and the mixture was reacted overnight at room temperature. The reaction solution was slowly poured into vigorously stirred deionized water, filtered, washed with cold deionized water, and dried under vacuum at 80 °C overnight to give a white solid product F. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ 11.59 (s, 1H), 10.79 (s, 1H), 9.33 (s, 1H), 8.59 (s, 1H), 7.48 –7.37 (m, 5H), 6.84 (t, J = 5.0 Hz, 2H), 5.31 (s, 2H), 1.47 (s, 9H). Compound F 1 HNMR spectrum as follows Figure 6 .

[0070] (6) Synthesis of compound G: Compound F (1 equiv.) and palladium / carbon (0.1 equiv.) were dissolved in methanol under a hydrogen atmosphere. Triethylamine (1.5 equiv.) was added, and the reaction was carried out overnight at 40 °C. After filtration, the methanol / triethylamine mixed solvent (25:1) was used. v / v Wash the product and let the filtrate stand overnight. Remove the solvent by vacuum distillation, add deionized water to the resulting solid, and freeze-dry to give a white solid product G. 1 H NMR (400 MHz, DMSO- d6, 298 K) δ 11.00 (s, 1H), 9.34 (s, 1H), 8.59(s, 1H), 6.78 (s, 1H), 6.55 (d, J = 3.4 Hz, 1H), 2.78 (dd, J = 13.0, 6.0 Hz, 6H),1.46 (s, 9H), 1.07 (t, J = 7.2 Hz, 9H). Compound G 1 H NMR spectrum as Figure 7 .

[0071] (6) Synthesis of compound H: Compound G (1 equiv.) was dissolved in deionized water. Dilute hydrochloric acid (1 N) was added, and stirred for 30 min. Filtration, deionized water washing, 80 °C vacuum drying overnight to get white solid product H. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ 11.38 (s, 1H), 9.33 (s, 1H), 8.60 (s, 1H), 6.78 (d, J = 19.7Hz, 2H), 1.47 (s, 9H). Compound H 1 H NMR spectrum as Figure 8 .

[0072] 2, guanidyl carbonyl pyrrole functionalization of amphiphilic resorcinol calixarene synthesis process as follows:

[0073] (1) Synthesis of compound I: resorcinol (1 equiv.) was dissolved in a mixed solvent of concentrated hydrochloric acid and methanol under nitrogen atmosphere. The ice-salt bath was used to reduce the temperature of the system to 20 °C, and 2,3-dihydrofuran (1 equiv.) was slowly added dropwise. The ice-salt bath was continued for 15 min. The ice-salt bath was removed, and the temperature was increased to 50 °C for 5 days of reaction. The system was cooled to room temperature, and filtration was performed. The filter cake was washed with methanol / water (1:1, v : v The obtained filter cake was suspended in deionized water, and ultrasonic treatment was performed for 10 min. Filtration was performed, and deionized water washing was performed. The filter cake was vacuum dried at 120 °C overnight to obtain white solid product I. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ 8.89 (s, 8H), 7.20 (s, 4H), 6.12(s, 4H), 4.32 (s, 4H), 4.18 (t,J = 7.9 Hz, 4H), 3.39 (d, J = 3.7 Hz, 8H), 2.06(dd, J = 14.3, 7.7 Hz, 8H), 1.35 – 1.27 (m, 8H). Compound I 1 H NMR spectrum as shown Figure 9 .

[0074] (2) Synthesis of compound J: Compound I (1 equiv.) and potassium carbonate (10 equiv.) were dissolved in DMA under a nitrogen atmosphere and stirred for 10 min. Bromochloromethane (20 equiv.) was added and the mixture was reacted at 50 °C for 5 days. The solvent was removed by vacuum distillation, and dilute hydrochloric acid (1 N) was added to the resulting solid. The mixture was sonicated for 10 min, filtered, washed with deionized water, and dried under vacuum at 80 °C overnight to obtain a white solid product J. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ 7.62 (s, 4H), 6.51 (s, 4H), 5.72 (d, J = 7.5 Hz, 4H), 4.56 (t, J = 7.9 Hz, 4H), 4.50 (t, J = 4.6 Hz, 4H), 4.39 (d, J = 7.6Hz, 4H), 3.51 (dd, J = 11.4, 5.8 Hz, 8H), 2.44 – 2.37 (m, 8H), 1.49 – 1.41 (m, 8H). Compound J's 1 H NMR spectrum as shown Figure 10 .

[0075] (3) Synthesis of compound K: Compound J (1 equiv.) was dissolved in dichloromethane under a nitrogen atmosphere. Triphenylphosphine (6 equiv.) and carbon tetrabromide (6 equiv.) were added sequentially, and the reaction was carried out overnight at room temperature. The solvent was removed by vacuum distillation, and the crude product was purified by column chromatography to obtain a white solid product K. 1 H NMR (400 MHz, CDCl3, 298 K) δ 7.18 (s,4H), 6.51 (s, 4H), 5.76 (d, J = 6.9 Hz, 4H), 4.78 (t, J = 7.9 Hz, 4H), 4.44 (d, J= 7.2 Hz, 4H), 3.57 (t, J = 5.9 Hz, 8H), 2.46 (dd, J = 15.1, 8.3 Hz, 8H), 1.95 (m, J = 14.4, 7.0 Hz, 8H). 13 C NMR (100 MHz, CDCl3, 298 K) δ 155.1, 138.1, 120.5,116.8, 99.5, 35.8, 33.9, 31.1, 28.4. HR-ESI-MS m / z: [M + H] + calcd for[C 44 H 45 O8Br4] + , 1018.9827; found, 1018.9851. Compound K 1 H NMR spectrum of compound K is shown in Figure 11 , compound K 13 C NMR spectrum of compound K is shown in Figure 12 ; HR-ESI-MS spectrum of compound K is shown in Figure 13 .

[0076] (4) Synthesis of compound L: Compound K (1 equiv.) and potassium phthalimide (6 equiv.) were dissolved in DMF under nitrogen atmosphere, and reacted at 40 °C for 1 day. The solvent was removed by distillation under reduced pressure, dichloromethane was added to the obtained solid, and deionized water was washed, and dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure. The obtained solid was dissolved in ethanol under nitrogen atmosphere, hydrazine hydrate was added, and reacted at 80 °C overnight. After cooling to room temperature, the solvent was removed by distillation under reduced pressure. Potassium hydroxide solution (2 M) was added, and ultrasonicated for 2 min, suction filtered, washed with deionized water, and dried at 80 °C under vacuum overnight to obtain white solid product L. 1 H NMR (400 MHz, DMSO- d 6, 298K) δ 7.58 (s, 4H), 6.52 (s, 4H), 5.72 (d, J = 7.5 Hz, 4H), 4.54 (t, J = 7.9 Hz,4H), 4.39 (d, J = 7.3 Hz, 4H), 2.67 (s, 8H), 2.39 (s, 8H), 1.39 (s, 8H). 13 C NMR(100 MHz, DMSO- d6, 298 K) δ 154.55, 138.73, 122.91, 116.91, 99.39, 42.01,36.62, 32.23, 26.83.HR-ESI-MS m / z: [M + H] + calcd for [C 44 H 53 O8N4] + , 765.3863; found, 765.3830. Compound L 1 H NMR spectrum as shown Figure 14 Compound L 13 The C NMR spectrum is as follows Figure 15 The HR-ESI-MS spectrum of compound L is as follows: Figure 16 .

[0077] (5) Synthesis of compound GCP-CA: Compound H (4 equiv.) and 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate (4.5 equiv.) were dissolved in DMF under a nitrogen atmosphere. [The remaining text appears to be incomplete and requires further context.] N , N - Diisopropylethylamine (5 equiv.), stirred for 40 min. Add L (1 equiv.), react at room temperature for 1 day. Remove the solvent by vacuum distillation, add EA to the resulting yellow oil, and stir for 10 min. Filter, wash with EA, and dry under vacuum at 70 °C for 6 h. Dissolve the resulting solid in dichloromethane, add trifluoroacetic acid, and react at room temperature for 2 h. Remove the solvent by vacuum distillation, add EA to the resulting yellow oil, and stir for 10 min. Filter, wash with EA, and dry under vacuum at 70 °C overnight to obtain the gray-green solid product GCP-CA. 1 H NMR (400 MHz, DMSO- d 6, 298 K) δ 12.28 (s, 4H), 11.13 (s, 4H), 8.38 (d, J = 41.8 Hz, 16H), 7.64 (s, 4H), 7.05 (s, 4H), 6.83 (s, 4H), 6.56 (s, 4H), 5.74 (d, J = 6.6 Hz,4H), 4.68 – 4.60 (m, 4H), 4.41 (d, J = 7.2 Hz, 4H), 3.39 (s, 8H), 1.58 (s, 8H). 13 C NMR (100 MHz, DMSO- d6, 298 K) δ 160.1, 159.9, 159.6, 159.5, 159.3, 155.6,154.7, 138.5, 133.3, 125.7, 122.8, 118.8, 117.1, 115.8, 112.8, 99.4, 36.5,31.4, 28.1, 26.8. HR-ESI-MS m / z: [M + Na] + calcd for [C 80 H 80 O 24 N 20 F 12 Na] + ,1955.5360; found, 1955.5381. Figure 17 For compound GCP-CA 1 H NMR spectrum; Figure 18 For compound GCP-CA 13 C NMR spectrum; Figure 19 The image shows the HR-ESI-MS spectrum of compound GCP-CA.

[0078] Example 2 Preparation of binary nanoparticles using guanidinocarbonylpyrrole functionalized amphiphilic resorcinarene calixarene-norfloxacin Figures 22-23 Guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticles are constructed by using guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene as the host and norfloxacin as the guest, through host-guest interactions.

[0079] The construction method is as follows: (1) At room temperature, guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene GCP-CA and norfloxacin (Nor) were dissolved in dimethyl sulfoxide (DMSO) to prepare GCP-CA mother liquor (concentration 18.75 mM) and Nor mother liquor (concentration 93.75 μM).

[0080] After mixing 40 μL of GCP-CA stock solution and 20 μL of Nor stock solution evenly, the mixture was quickly injected into 2940 μL of deionized water to obtain binary nanoparticles. Solution A has host and guest concentrations of 0.25 mM and 0.625 μM, respectively. For example... Figures 20-21 As shown, the constructed guanidinocarbonylpyrrole-functionalized amphiphilic resorcinol calixarene-norfloxacin binary nanoparticles have uniform size and a hydrated diameter of approximately 458 nm.

[0081] Take 20 μL GCP-CA mother liquor, 10 μL Nor mother liquor mixed evenly and quickly injected into 2970 μL deionized water to obtain binary nanoparticle (GCP-CA-Nor) solution B, at this time the host and guest concentrations are 0.125 mM and 0.3125 μM respectively. As shown in the figure, the guanidyl carbonyl pyrrole functionalized amphiphilic resorcinarene nanoparticle size is uniform, and the hydration diameter is about 531 nm. Performance test examples As a control, the nanoparticle construction method of the host or the guest alone is as follows: Take 40 μL GCP-CA mother liquor and quickly inject into 2960 μL deionized water to obtain GCP-CA nanoparticle solution C, at this time the host concentration is 0.25 mM.

[0082] Take 20 μL GCP-CA mother liquor and quickly inject into 2980 μL deionized water to obtain GCP-CA nanoparticle solution D, at this time the host concentration is 0.125 mM.

[0083] Take 20 μL Nor mother liquor and quickly inject into 2980 μL deionized water to obtain Nor nanoparticle solution E, at this time the guest concentration is 0.625 μM.

[0084] Take 10 μL Nor mother liquor and quickly inject into 2990 μL deionized water to obtain Nor nanoparticle solution F, at this time the guest concentration is 0.3125 μM.

[0085] S. aureus The GCP-CA prepared in Example 1 and the carbonyl pyrrole functionalized amphiphilic resorcinarene-norfloxacin binary nanoparticle (GCP-CA-Nor) constructed in Example 2 were tested for antibacterial performance: the antibacterial effects of GCP-CA, Nor, on Staphylococcus aureus (ATCC 6538) and Escherichia coli (ATCC 8739) were evaluated by plate counting method. coli E. S. aureus

[0086] 1. Staphylococcus aureus antibacterial test The test groups are as follows: Control group: no any drug is applied; GCP-CA group: GCP-CA nanoparticle solution C (concentration is 0.25 mM) Nor group: Nor nanoparticle solution E (concentration is 0.625 μM): Group: binary nanoparticle (GCP-CA-Nor) solution B (concentration of host and guest is 0.125 mM and 0.3125 μM respectively). ​​​​Solution A; wherein the concentrations of the host and guest are 0.25 mM and 0.625 μM, respectively.

[0087] Test method: Staphylococcus aureus ( E. coli ATCC 6538 bacterial suspension diluted to a concentration of 2.0 × 10⁻⁶. 6 CFU / mL; Take 200 μL of bacterial suspension and add 200 μL of the corresponding drug solution for each experimental group (the control group is added with an equal volume of deionized water); After incubation at 37 ℃ for 2 h, dilute the drug-bacterial mixture 100 times, take 50 μL and spread it on MH agar plates, incubate overnight at 37 ℃, and count the bacterial colonies on the agar plates to calculate the bacterial survival rate. Three parallel groups were set up for each sample.

[0088] 2. Escherichia coli antibacterial test The experimental groups are as follows: Control group: No drugs were administered; GCP-CA group: GCP-CA nanoparticle solution D (concentration of 0.125 mM) Nor group: Nor nanoparticle solution F (concentration 0.3125 μM): Group: Binary nanoparticles ( Solution B; the host and guest concentrations were 0.125 mM and 0.3125 μM, respectively.

[0089] Test method: Escherichia coli (E. coli) Figure 24 The bacterial suspension (ATCC 8739) was diluted to a concentration of 2.0 × 10⁻⁶. 6 CFU / mL; Take 200 μL of bacterial suspension and add 200 μL of the corresponding drug solution for each experimental group (the control group is added with an equal volume of deionized water); After incubation at 37 ℃ for 2 h, dilute the drug-bacterial mixture 100 times, take 50 μL and spread it on MH agar plates, incubate overnight at 37 ℃, and count the bacterial colonies on the agar plates to calculate the bacterial survival rate. Three parallel groups were set up for each sample.

[0090] The results are as follows Figure 25 As shown, when GCP-CA or Nor is co-incubated with bacteria alone, they exhibit very weak antibacterial activity due to their concentrations being far below the MIC value, and a large number of bacteria still survive in the culture dish. However, when the same concentration of GCP-CA and Nor is used to prepare ( When these two act together on bacteria, It exhibited excellent synergistic antibacterial activity, with almost no colonies visible in the culture dish.

[0091] Figure 26Statistics on bacterial survival rates show that after incubation with GCP-CA or Nor alone, approximately 70% of the bacteria still survive. However, with... After incubation, the bacterial survival rate was close to 0. This was observed with GCP-CA, Nor, and [other bacteria / organisms]. After co-incubation, the morphology of the bacteria is as follows: ​ As shown, the bacteria in the control group (without any drugs applied) had smooth, flat surfaces, clear boundaries, and full morphology. The bacteria in the GCP-CA and Nor groups showed slight boundary blurring and collapse. The bacterial membranes in the group showed obvious wrinkles and damage, and bacterial collapse caused by leakage of contents could be observed, while the boundaries were blurred and the bacteria were stuck together.

[0092] The above results indicate that GCP-CA and Nor have excellent synergistic antibacterial effects and can exhibit very good in vitro antibacterial activity at concentrations far below their respective MIC values.

[0093] Specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various changes or modifications within the scope of the claims. This does not affect the essence of the present invention. Unless otherwise specified, the embodiments and features described in these embodiments can be arbitrarily combined with each other.

Claims

1. A guanidinylcarbonyl pyrrole compound, having a structure as shown in the following formula: 。 2. The method for synthesizing the guanidinocarbonylpyrrole compound as described in claim 1, characterized in that, comprising the following steps: (a) reacting a pyrrole with a carboxylic acid derivative to form a compound A, having a structure as shown in the following formula: ； (b) dissolving the compound A in a halogenated hydrocarbon solvent under inert gas protection to obtain a compound A solution; adding a solution of an alkoxy alkali metal compound dropwise into the compound A solution to react, forming a compound B, having a structure as shown in the following formula: ； (c) performing a formylation reaction on the compound B in the presence of a Vilsmeier-Haack reagent to form a compound C, having a structure as shown in the following formula: ； (d) oxidizing the compound C in the presence of an oxidizing agent to form a carboxylic acid compound D, having a structure as shown in the following formula: ； (e) reacting a guanidine or a guanidine salt with a protection reagent under alkaline conditions to form a protected guanidine compound E, having a structure as shown in the following formula: ； (f) performing a condensation reaction on the carboxylic acid compound D and the protected guanidine compound E in the presence of a coupling agent and an organic base to form a compound F, having a structure as shown in the following formula: ； (g) performing a catalytic hydrogenation deprotection reaction on the compound F in the presence of a catalyst and a base in a reducing atmosphere to form a compound G, having a structure as shown in the following formula: ； (h) performing a final acidification reaction on the compound G under acidic conditions to obtain the guanidinylcarbonyl pyrrole compound.

3. A guanidinocarbonyl pyrrole functionalized amphiphilic resorcinarene calixarene (GCP-CA) characterized in that, having a structure as shown in the following formula (1): Formula (1); wherein the number of carbon atoms n in the alkyl chain is 1-10.

4. The guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene GCP-CA according to claim 3, characterized in that, The guanidinylcarbonyl pyrrole functionalized amphiphilic resorcinol calixarene has a structure as shown in the following formula: 。 5. A method for preparing a guanidylcarbonyl pyrrole functionalized amphiphilic catechol calixarene GCP-CA, characterized in that, comprising the following steps: S1, reacting a polyphenol compound with a cyclization reagent under acidic conditions to form a cyclic compound I, having a structure as shown in the following formula (2): Formula (2); S2, reacting the cyclic compound I with a halomethylation reagent under alkaline conditions to obtain a compound J, having a structure as shown in the following formula (3): Formula (3); S3, performing a halogen exchange reaction on the compound J in the presence of a bromination reagent to obtain a bromomethyl compound K, having a structure as shown in the following formula (4): Formula (4); S4, reacting the bromomethyl compound K with an amine source to form an amine compound L, having a structure as shown in the following formula (5): Formula (5); S5, performing a condensation reaction on the amine compound L and the guanidinylcarbonyl pyrrole compound of claim 1 in the presence of a coupling agent and an organic base, and removing the protection group in the presence of trifluoroacetic acid to obtain the guanidinylcarbonyl pyrrole functionalized amphiphilic resorcinol calixarene, denoted as GCP-CA.

6. The preparation method according to claim 5, characterized in that, In step S1, the polyphenol compound is resorcinol; the cyclization reagent is 2,3-dihydrofuran; the molar ratio of resorcinol to 2,3-dihydrofuran is 1:0.9-1.1; the acidic conditions are provided by concentrated hydrochloric acid; the reaction is performed at a temperature ranging from -20 ℃ to 50 ℃, and the reaction time is 12 hours to 5 days. And / or, in step S2, the basic condition is provided by potassium carbonate, the molar ratio of compound I to potassium carbonate is 1:9-11; the halomethylating agent is bromochloromethane; the molar ratio of compound I to bromochloromethane is 1:19-21; the reaction temperature is 45-55℃, and the reaction time is 4-6 days. And / or, in step S3, the brominating agent is a combination of triphenylphosphine and carbon tetrabromide; the molar ratio of compound J to triphenylphosphine is 1:5-7; the molar ratio of compound J to carbon tetrabromide is 1:5-7; the reaction is carried out at 20-30℃, and the reaction time is 10-24 hours.

7. The preparation method according to claim 5, characterized in that, Step S4 comprises the following steps: S4a, reacting compound K with a phthalimide salt to obtain a protected amine intermediate; the molar ratio of compound K to the phthalimide salt is 1:5-7; the reaction temperature of this step is 35-45℃, and the reaction time is 20-28 hours; S4b, deprotecting the protected amine intermediate using a hydrazine reagent to obtain the amine compound L; the deprotection reaction is carried out at 70-90℃, and the reaction time is 10-24 hours; In S4a, the phthalimide salt is potassium phthalimide; and / or the hydrazine reagent is hydrazine hydrate; And / or, step S5 comprises the following steps: S5a, activating the carboxyl group of the guanidinocarbonyl pyrrole compound in the presence of a coupling agent and an organic base; the molar ratio of the carboxyl group of the guanidinocarbonyl pyrrole compound, the coupling agent and the organic base is 1:1.0-1.2:1.2-1.5, and the activation time is 30-60 minutes; S5b, condensing the activated carboxyl group of the guanidinocarbonyl pyrrole compound with the amine compound L to obtain a protected calixarene derivative; the molar ratio of the carboxyl group of the guanidinocarbonyl pyrrole compound to the amine compound L is 3.5-4.5:1; the condensation reaction is carried out at 20-30℃, and the reaction time is 20-28 hours; S5c, removing the protecting group in the presence of trifluoroacetic acid to obtain the final product guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene GCP-CA; the deprotection reaction is carried out at 20-30℃, and the reaction time is 1-3 hours.

8. A preparation method of a binary nanoparticle based on guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene, comprising the following steps: Step 1, dissolving the guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene GCP-CA of claim 3 or 4 and norfloxacin in organic solvents respectively to obtain GCP-CA mother liquor and norfloxacin mother liquor; Step 2, mixing the GCP-CA mother liquor and the norfloxacin mother liquor to obtain a host-guest mixture; Step 3, injecting the host-guest mixture into an aqueous medium, uniformly mixing to obtain the binary nanoparticle solution based on guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene.

9. A binary nanoparticle based on guanidinocarbonyl pyrrole functionalized amphiphilic resorcinol calixarene obtained by the preparation method of claim 8.

10. Use of a guanidylcarbonyl pyrrole-based functionalized amphiphilic resorcinarene-based binary nanoparticle according to claim 9 for the preparation of an antibacterial medicament.

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