CN110787298B - Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial - Google Patents
Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial Download PDFInfo
- Publication number
- CN110787298B CN110787298B CN201911016279.1A CN201911016279A CN110787298B CN 110787298 B CN110787298 B CN 110787298B CN 201911016279 A CN201911016279 A CN 201911016279A CN 110787298 B CN110787298 B CN 110787298B
- Authority
- CN
- China
- Prior art keywords
- star
- antibiotics
- hollow
- nano material
- shaped hollow
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/429—Thiazoles condensed with heterocyclic ring systems
- A61K31/43—Compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula, e.g. penicillins, penems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
- A61K31/545—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
- A61K31/546—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine containing further heterocyclic rings, e.g. cephalothin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/65—Tetracyclines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
Abstract
The invention discloses preparation and application of a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis. The star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis takes hollow polydopamine as a core and dendritic polyamide as an arm; the preparation method comprises the following steps: (1) synthesizing hollow polydopamine nanoparticles; (2) hollow poly-dopamine nanoparticles modified by azide groups; (3) and (3) reacting the azide group modified hollow polydopamine nano particles prepared in the step (2) with a dendritic polyamide aqueous solution to prepare the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis. The star-shaped hollow nano material (h-PDA @ PAMAM) with the hollow polydopamine as the core and the dendritic polyamide as the arm can load various antibiotics, so that the diversity of the nano hollow material is greatly increased, and the application range of the nano hollow material in the field of biological materials is widened.
Description
Technical Field
The invention belongs to the field of biomedical engineering materials, and particularly relates to preparation and application of a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis.
Background
Diseases caused by bacterial infections are one of the causes of death. Despite great efforts in the development of new drugs, diseases induced by bacterial infections still have a significant impact on human health and economy. In recent years, due to advances in research and diagnostic techniques, it has been found that the focus of refractory bacterial infections is largely dominated by a source of infection, the so-called bacterial biofilm, whose formation and persistence have considerable effects on the health of the patient, e.g. many refractory infections are difficult to eradicate, antibiotic therapy is difficult to achieve with significant efficacy, and can lead to chronic or recurrent infections. In fact, the resistance of the bacteria in the biofilm state is significantly increased compared to the resistance of the individual bacteria in the planktonic state, and can be resistant to attacks by adverse factors such as bacteriophages in nature, amoeba, antiseptics and antibiotics used by humans. This is due to the fact that after a certain degree of biofilm formation, the bacteria in the biofilm secrete a large amount of viscous extracellular matrix, which, like armor, generally encapsulates the bacteria and thus creates an incredible resistance to different antibiotics or antimicrobials. Therefore, there are several clinical challenges associated with biofilm infection, including those related to impaired wound healing, chronic inflammation, antibiotic resistance, and the spread of infectious diseases, and there is a strong need for new therapeutic concepts and methods that are effective against biofilms and biofilm-induced infections.
Biofilm researchers have established that most bacteria follow the main phase of the bacterial growth cycle that growth in the biofilm mode is achieved by the alternative switching of biofilm growth and planktonic growth through highly regulated and specific expression of genes. In recent years, the biological signaling molecule, nitric oxide, a diatomic free radical, has been identified as a key regulator of biofilm dispersion, and endogenous NO has been found to have a dispersion-inducing effect on mature biofilms. Molecular analysis suggests that NO triggers intracellular second messenger cyclic diguanylic acid (di-GMP) to reverse activate a series of effectors that lead to biofilm dispersion. Nageshwar et al found that the use of low concentrations of NO donor material, safe and without toxic side effects, was able to induce biofilm bacteria to revert to a range sensitive to antibiotics or antibacterial agents (chem. Furthermore, in a recent report, Louise et al can effectively eliminate bacteria from animal Infection models without much harm to the host by using donor materials that release NO gas under specific conditions (Infection and Immunity p.2705-2713). All of these combined results thus reveal an anti-biofilm strategy, i.e. the eradication of bacterial biofilm infections using low concentrations of NO donor compounds in combination with antibiotics.
However, a single NO donor shows poor antimicrobial efficacy in biological applications, and the antimicrobial efficacy is not significant, especially against the increasingly severe bacterial biofilm phenomenon, which greatly limits any potential applications. In order to overcome the problems, the NO donor material is combined with various antibacterial means (such as antibacterial drugs), so that the dynamic process of the bacterial biofilm formation process, namely the effective inhibition of NO, and the dissipation of the formed biofilm are realized, and the requirements of the antibacterial drugs on the bactericidal effect and the like are obviously improved.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the invention aims to provide a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis, namely a star-shaped hollow nano material (h-PDA @ PAMAM) taking hollow polydopamine as a core and dendritic polyamide as an arm.
The invention also aims to provide a preparation method of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis.
The invention further aims to provide application of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis.
The invention also provides a star-shaped hollow nano particle (h-PDA @ PAMAM/PRL/NONOate) which takes hollow polydopamine as a core and dendritic polyamide as an arm and can simultaneously release Nitric Oxide (NO) and antibiotic Piperacillin (PRL).
The purpose of the invention is realized by the following technical scheme:
a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis takes hollow polydopamine as a core and dendritic polyamide as an arm, and the structural formula is shown in figure 6.
The preparation method of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis comprises the following steps:
(1) synthesis of hollow Polydopamine nanoparticles (pDA HNP)
Preparing silicon dioxide nano particles (SiNP), dispersing the silicon dioxide nano particles in a buffer solution, adding dopamine hydrochloride, stirring and reacting for 24-48 hours to obtain poly-dopamine-coated silicon dioxide nano particles (pDA @ SiNP); dispersing pDA @ SiNP into a mixed solution of hydrofluoric acid and ammonium fluoride, and continuously reacting for 24-48 h to obtain hollow polydopamine nano particles (pDA HNP);
(2) hollow poly-dopamine nanoparticle (pDA-N) modified by azide group3 HNP)
Reacting azidoacetic acid (N)3-CH2-COOH) is dissolved in an organic solvent, and then 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl) and N-hydroxysuccinimide (NHS) are added for activation for 30 min-4 h; dispersing the pDA HNP prepared in the step (1) in pure water, then adding the pure water into the azidoacetic acid mixed solution after the activation is finished, and reacting at room temperature for 12-24 h to prepare the azido group modified hollow polydopamine nano particle (pDA-N)3 HNP)；
(3) Synthesis of star-shaped hollow nano material (h-PDA @ PAMAM) with hollow polydopamine as core and dendritic polyamide as arm
Reference is made to the patent: a nitric oxide loaded cationic polymer, a preparation method and application thereof (patent number is ZL 201610356643.9) for preparing alkynyl-containing tri-generation dendritic Polyamide (PAMAM);
the pDA-N prepared in the step (2)3Dispersing HNP in organic solvent, introducing protective gas, adding dendritic Polyamide (PAMAM) water solution, and adding copper sulfate pentahydrate (CuSO) with a certain mass4·5H2O) and sodium ascorbate (NaVc), continuously introducing protective gas, and reacting at 40-75 ℃ for 24-48 h to prepare the star-shaped hollow nano material (h-PDA @ PAMAM) taking hollow polydopamine as a core and dendritic polyamide as an arm.
Further, in the step (1):
the silica nanoparticles are prepared by the following steps: uniformly mixing 25% ammonia water, absolute ethyl alcohol and pure water, adding 99% tetraethyl orthosilicate (TEOS) to change a system solution from transparent to white, continuously stirring for 5-10 h, centrifuging to remove a suspension, and alternately washing with ethanol and pure water to obtain silicon dioxide nanoparticles (SiNP); the volume ratio of the ammonia water to the absolute ethyl alcohol to the pure water to the ethyl orthosilicate is 1: 10-30: 2-5: 0.5 to 2;
the buffer solution is a Tris buffer solution with the pH value of 7.5-9;
the mass ratio of the silicon dioxide nanoparticles to the dopamine hydrochloride is 1: 1-5;
the mass volume ratio of the pDA @ SiNP to the mixed solution of hydrofluoric acid and ammonium fluoride is 50-100 mg: 5-10 mL;
the molar concentration ratio of hydrofluoric acid to ammonium fluoride in the mixed solution of hydrofluoric acid and ammonium fluoride is 1:2 to 5.
Further, in the step (2):
the mole ratio of the hollow polydopamine nanoparticles (pDA HNP), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl), N-hydroxysuccinimide (NHS) and azidoacetic acid is (1: 1) - (10): 1-10: 1-10;
the organic solvent is at least one of N-N-Dimethylformamide (DMF), dimethyl sulfoxide, methanol and the like; adding 1-5 g of azido acetic acid into each 10mL of organic solvent;
the pure water is calculated by adding 0.1-5 g of azido acetic acid into every 10 mL.
Further, in the step (3):
the pDA-N3The mass ratio of HNP to dendritic polyamide is 1: 50-100;
the PAMAM and the CuSO4·5H2The molar ratio of O to NaVc is 1: 7-14: 1-3: 3-9;
the organic solvent is at least one of dimethyl sulfoxide (DMSO), DMF, tetrahydrofuran and the like, and 50-100 mg of pDA-N is added into 10mL of DMSO3An HNP meter; the amount of the pure water is calculated by adding 1-3 g of PAMAM into every 10 mL;
the protective gas is nitrogen, and the dendritic Polyamide (PAMAM) aqueous solution is added after the protective gas is introduced for 10-30 min.
The star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis can be used for loading nitric oxide and antibiotics and used as an antibacterial drug. Besides PRL, the antibiotic can also load other antibiotics such as vancomycin, cefixime, ampicillin, tetracycline and norfloxacin.
Further, the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis is used for loading nitric oxide and piperacillin, and the star-shaped nano material capable of releasing NO and antibiotics for synergistic antibiosis is prepared by the following specific steps:
a. dispersing the star-shaped hollow nano material (h-PDA @ PAMAM) capable of simultaneously loading NO and antibiotics for synergistic antibiosis into pure water at room temperature, then adding a certain mass of antibiotics, and continuously stirring for 24-48 h to obtain the star-shaped hollow nano material loaded with the antibiotics; the piperacillin, cefixime, vancomycin, ampicillin, tetracycline or norfloxacin;
b. drying the antibiotic-loaded star-shaped hollow nano material, dispersing the dried star-shaped hollow nano material in anhydrous methanol, tetrahydrofuran or acetonitrile, adding sodium methoxide, stirring for 10-30 min, placing the mixture in a high-pressure reaction kettle, sealing the high-pressure reaction kettle, introducing high-purity nitrogen to remove air in the high-pressure reaction kettle, introducing NO gas, and reacting at room temperature for 3-7 days to prepare the NO and antibiotic-loaded star-shaped nano material for synergistic antibiosis.
Further, in the above-mentioned case,
the mass ratio of the antibiotic-loaded star-shaped hollow nano material to the antibiotic is 1: 1-10; the pure water is calculated by adding 50-100 mg of h-PDA @ PAMAM into every 10 mL;
the mass ratio of the antibiotic-loaded star-shaped hollow nano material to sodium methoxide is 1: 5-20; the absolute methanol is calculated by adding 0.1-0.5 g of sodium methoxide into every 10mL of absolute methanol;
introducing high-purity nitrogen to maintain the reaction kettle at 10-20 psi for 5-15 min, and removing air in the reaction kettle; the pressure of the high-purity nitrogen is 10-20 psi, and the pressure of the NO gas is 40-80 psi.
In the invention, the preparation method of the absolute ethyl alcohol comprises the following operation steps: adding calcium hydride into methanol, stirring for 6-24 hours, and then distilling at normal pressure to obtain anhydrous ethanol, wherein the addition amount of the calcium hydride is 1-2 g per 500mL of ethanol;
the preparation method of the anhydrous methanol comprises the following operation steps: adding calcium hydride into methanol, stirring for 6-24 hours, and then distilling at normal pressure to obtain anhydrous methanol, wherein the addition amount of the calcium hydride is 1-2 g per 500mL of methanol;
the room temperature is 5-35 ℃.
The synthetic route of the star-shaped nanometer material capable of releasing NO and antibiotics for synergistic antibiosis is shown in figure 7.
The star-shaped nano material (h-PDA @ PAMAM/PRL/NONONAte) capable of releasing NO and antibiotics and used for synergistic antibacterial can be used as a novel antibacterial drug.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the high-generation dendritic polyamide is used as an NO donor, so that the NO loading is greatly improved, the effect on a bacterial biofilm is obviously inhibited, and the negatively charged bacteria can be well adsorbed and have a certain killing effect;
(2) the star-shaped hollow nano material (h-PDA @ PAMAM) with the hollow polydopamine as the core and the dendritic polyamide as the arm can load various antibiotics, so that the diversity of the nano hollow material is greatly increased, and the application range of the nano hollow material in the field of biological materials is widened;
(3) compared with single NO or PRL, the star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONAte) loaded with NO and PRL together have more obvious effect on resisting planktonic bacteria;
(4) the design and construction of using star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONAte) as NO donors realize high-efficiency loading of NO and quick dissipation of biological membranes, and induce stubborn bacteria in the biological membranes to recover to planktonic bacteria sensitive to antibiotics, and at the moment, a large amount of PRL is released from the hollow nanoparticles to quickly improve local concentration, so that the synergistic effect of two released medicines in the biological field is realized to resist the problem of drug-resistant bacteria caused by the existence of the biological membranes;
(5) the star-shaped hollow nano-particle (h-PDA @ PAMAM/PRL/NONOnoate)) which takes the hollow polydopamine as the core and the dendritic polyamide as the arm and can simultaneously release nitric oxide and the antibiotic piperacillin has the advantages of effectively inhibiting the growth and the reproduction of bacteria and fungi, having obvious inhibiting effects on common pathogenic bacteria, dermatophytes, wound infectious bacteria and the like, having the functions of promoting wound healing, diminishing inflammation and the like, and providing support for the application of the star-shaped hollow nano-particle in the preparation of biomedical engineering materials.
Drawings
FIG. 1 is a TEM image of hollow polydopamine nanoparticles (pDA HNPs);
FIG. 2 is a diagram showing potential changes in the synthesis process of star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONAte);
FIG. 3 is a dynamic process of release of NO and PRL by star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONOATE) under simulated human physiological conditions;
FIG. 4 is a graph showing the bactericidal effect of star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONOATE) on E.coli in a floating state compared to other materials;
FIG. 5 shows the bactericidal effect of h-PDA @ PAMAM, h-PDA @ PAMAM/PRL and h-PDA @ PAMAM/PRL/NONOATE;
FIG. 6 is a structural formula of a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial of the present invention;
FIG. 7 is a synthetic route of the star-shaped nano material capable of releasing NO and antibiotics for synergistic antibacterial action according to the present invention.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto.
The preparation of anhydrous methanol as described in the following examples follows the following operating steps: adding calcium hydride into methanol, stirring for 6-24 hours, and then distilling at normal pressure to obtain anhydrous methanol, wherein the addition amount of the calcium hydride is 1-2 g per 500mL of methanol.
The preparation of the anhydrous ether described in the following examples follows the following operating steps: adding absolute ethyl alcohol into ethyl ether, stirring for 6-24 hours, and then distilling at normal pressure to obtain the absolute ethyl alcohol, wherein the addition amount of the calcium hydride is 1-2 g per 500mL of the ethyl alcohol.
Example 1
Synthesis of hollow Polydopamine nanoparticles (pDA HNP)
a. Uniformly mixing 25% ammonia water, absolute ethyl alcohol and pure water in certain volume, adding 99% tetraethoxysilane in certain volume concentration to change the system solution from transparent to white, continuously stirring for 10h, centrifuging at 8000rpm to remove the suspension, and alternately washing with ethanol and pure water for 4 times to obtain silicon dioxide nano particles (SiNP); the volume ratio of the ammonia water, the ethanol, the pure water and the TEOS is 1: 30: 5: 2;
b. washing the SiNP obtained in the step a by using 10mM Tris buffer solution (pH9) for 3 times, then dispersing the SiNP into 50mL Tris buffer solution again, then adding dopamine hydrochloride with a certain mass, stirring and reacting for 48 hours, washing by pure water centrifugation (8000rpm) for 2-3 times, and finally obtaining the polydopamine-coated silicon dioxide nanoparticles (pDA @ SiNP); the mass ratio of the silicon dioxide nano particles (SiNP) to the dopamine hydrochloride is 1: 5;
c. and c, dispersing 100mg of the pDA @ SiNP nano particles obtained in the step b into 10mL of mixed solution of hydrofluoric acid and ammonium fluoride, continuing to react for 48h, and then centrifuging (8000rpm) to wash with pure water for 3 times to finally obtain hollow polydopamine nano particles (pDA HNP). The molar concentration ratio of the hydrofluoric acid to the ammonium fluoride is 1: 5.
Example 2
Synthesis of hollow Polydopamine nanoparticles (pDA HNP)
d. Uniformly mixing 25% ammonia water, absolute ethyl alcohol and pure water in certain volume, adding 99% tetraethoxysilane in certain volume concentration to change the system solution from transparent to white, continuously stirring for 5h, centrifuging at 5000rpm to remove the suspension, and alternately washing with ethanol and pure water for 2 times to obtain silicon dioxide nano particles (SiNP); the volume ratio of the ammonia water, the ethanol, the pure water and the TEOS is 1:10:2: 0.5;
e. b, washing the SiNP obtained in the step a with 10mM Tris buffer solution (pH7.5) for 2-3 times, then dispersing the SiNP into 30mL Tris buffer solution again, then adding dopamine hydrochloride with a certain mass, stirring and reacting for 24 hours, washing with pure water in a centrifugal mode (5000rpm) for 2 times, and finally obtaining poly-dopamine-coated silicon dioxide nanoparticles (pDA @ SiNP); the mass ratio of the silicon dioxide nano particles (SiNP) to the dopamine hydrochloride is 1: 1;
f. and c, dispersing 50mg of the pDA @ SiNP nano particles obtained in the step b into 5mL of mixed solution of hydrofluoric acid and ammonium fluoride, continuing to react for 24h, and then centrifuging (5000rpm) to wash with pure water for 2 times to finally obtain hollow polydopamine nano particles (pDA HNP). The molar concentration ratio of the hydrofluoric acid to the ammonium fluoride is 1: 2.
Example 3
Hollow poly-dopamine nanoparticle (pDA-N) modified by azide group3 HNP)
Reacting azidoacetic acid (N)3-CH2-COOH) was dissolved in N-N-Dimethylformamide (DMF) and then activated for 30min by the sequential addition of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl) and N-hydroxysuccinimide (NHS) (meaning a total of 30min after addition of EDC. HCl and NHS). The same applies below); after the activation, the hollow poly dopamine nanoparticles obtained in example 1 were dispersed in pure water, and then slowly added to the azido-acetic acid mixed solution, followed by reaction at room temperature for 12 hours. Centrifugation (5)000rpm) was washed 2 times with pure water (after centrifugation, pure water was resuspended and centrifuged again to remove pure water, then fresh pure water was added for dispersion and centrifuged again, and this step was repeated several times, and this step was collectively referred to as pure water centrifugal washing. The same below) to finally obtain the hollow polydopamine nano-particle (pDA-N)3HNP). The molar ratio of the hollow polydopamine nanoparticles (pDA HNP), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl), N-hydroxysuccinimide (NHS) and azidoacetic acid is 1: 1: 1: 1; the N, N-dimethylformamide is calculated by adding 1g of azidoacetic acid into every 10 milliliters; the pure water is calculated by adding 0.1g of azidoacetic acid into every 10 mL.
Example 4
Hollow poly-dopamine nanoparticle (pDA-N) modified by azide group3 HNP)
Reacting azidoacetic acid (N)3-CH2-COOH) in N-N-Dimethylformamide (DMF), followed by the sequential addition of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl), N-hydroxysuccinimide (NHS) for 4h activation; after the activation, the hollow poly dopamine nanoparticles obtained in example 2 were dispersed in pure water, and then slowly added to the azido-acetic acid mixed solution, followed by reaction at room temperature for 24 hours. Washing with pure water at 8000rpm for 3 times to obtain hollow polydopamine nanoparticles (pDA-N)3HNP). The molar ratio of the hollow polydopamine nanoparticles (pDA HNP), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl), N-hydroxysuccinimide (NHS) and azidoacetic acid is 1:10: 10: 10; the N, N-dimethylformamide is calculated by adding 5g of azidoacetic acid into every 10 milliliters; the pure water is calculated by adding 5g of azidoacetic acid into every 10 mL.
Example 5
Synthesis of alkynyl-containing tri-generation dendritic Polyamide (PAMAM)
Reference is made to the patent: a nitric oxide loaded cationic polymer, a preparation method and application thereof (patent number ZL 201610356643.9) which are specifically obtained by implementation 4.
Example 6
Synthesis of alkynyl-containing tri-generation dendritic Polyamide (PAMAM)
Reference is made to the patent: a nitric oxide loaded cationic polymer, a preparation method and application thereof (patent No. ZL 201610356643.9) as well as a product obtained by the specific implementation 5.
Example 7
Synthesis of star-shaped hollow nano material (h-PDA @ PAMAM) with hollow polydopamine as core and dendritic polyamide as arm
50mg of pDA-N obtained in example 33Dispersing HNP in dimethyl sulfoxide (DMSO), introducing nitrogen for 20min, and slowly adding the aqueous solution of dendritic Polyamide (PAMAM) obtained in example 5; pDA-N3The mass ratio of HNP to dendritic polyamide is 1: 50. Then adding a certain mass of copper sulfate pentahydrate (CuSO)4·5H2O) and sodium ascorbate (NaVc), continuously introducing nitrogen, and reacting for 24h at 60 ℃. And finally, centrifuging the reaction solution (6000rpm) and washing with pure water for 2 times to obtain the star-shaped hollow nano material (h-PDA @ PAMAM) taking the hollow polydopamine as a core and the dendritic polyamide as an arm. The PAMAM and the CuSO4·5H2The molar ratio of O to NaVc is 1:10:2: 6; the DMSO is added with 50mg of pDA-N per 10mL3An HNP meter; the amount of the pure water is calculated by adding 2g of PAMAM in every 10 mL.
Example 8
Synthesis of star-shaped hollow nano material (h-PDA @ PAMAM) with hollow polydopamine as core and dendritic polyamide as arm
100mg of pDA-N obtained in example 43Dispersing HNP in dimethyl sulfoxide (DMSO), introducing nitrogen for 30min, and slowly adding the aqueous solution of dendritic Polyamide (PAMAM) obtained in example 6; pDA-N3The mass ratio of HNP to dendritic polyamide is 1: 100. then adding a certain mass of copper sulfate pentahydrate (CuSO)4·5H2O) and sodium ascorbate (NaVc), introducing nitrogen continuously, and reacting for 48h at 75 ℃. And finally, centrifuging the reaction solution (8000rpm) and washing with pure water for 3 times to obtain the star-shaped hollow nano material (h-PDA @ PAMAM) taking the hollow polydopamine as a core and the dendritic polyamide as an arm. The PAMAM and the CuSO4·5H2The molar ratio of O to NaVc is 1:14:3: 9; the DMSO is added with 100mg of pDA-N per 10mL3An HNP meter; the amount of the pure water is calculated by adding 3g of PAMAM in each 10 mL.
Example 9
Antibiotic vancomycin (NVA), Cefixime (CFM) and Nitric Oxide (NO) gas co-loading is synthesis of final product
(1) a, dispersing the star-shaped hollow nano material (h-PDA @ PAMAM) obtained in a certain mass in certain volume of pure water at room temperature of 25 ℃, then slowly adding PRL with a certain mass, continuously stirring for 48h, finally centrifuging (8000rpm) and washing with pure water for 3 times, and finally obtaining the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/PRL). The mass ratio of the h-PDA @ PAMAM to the PRL is 1: 10; the pure water is calculated by adding 100mg of h-PDA @ PAMAM into every 10 mL;
b. dispersing the star-shaped hollow nano material (h-PDA @ PAMAM) obtained in a certain mass in pure water with a certain volume at room temperature of 25 ℃, slowly adding vancomycin (NVA) with a certain mass, continuously stirring for 48h, and finally centrifuging (8000rpm) to wash with pure water for 3 times to finally obtain the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/NVA). The mass ratio of the h-PDA @ PAMAM to the NVA is 1: 10; the pure water is calculated by adding 100mg of h-PDA @ PAMAM into every 10 mL;
c. dispersing the star-shaped hollow nano material (h-PDA @ PAMAM) obtained in a certain mass in pure water with a certain volume at the room temperature of 25 ℃, then slowly adding Cefixime (CFM) with a certain mass, continuously stirring for 48h, finally centrifuging (8000rpm) and washing with pure water for 3 times, and finally obtaining the CFM-loaded star-shaped hollow nano material (h-PDA @ PAMAM/CFM). The mass ratio of the h-PDA @ PAMAM to the PRL is 1: 10; the pure water is calculated by adding 100mg of h-PDA @ PAMAM into every 10 mL;
(2) and (2) drying the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/PRL) obtained in the step (1), dispersing the dried star-shaped hollow nano material in absolute methanol, adding sodium methoxide, stirring and stabilizing for 30min, placing the mixture in a high-pressure reaction kettle, sealing, and detecting the air tightness. The reaction kettle (20psi) is maintained by high-purity nitrogen for 15min to remove air in the reaction kettle, and then NO gas (80psi) is introduced to react for 7 days at room temperature. After the reaction is finished, discharging NO by high-purity nitrogen with the pressure of 20psi, continuously maintaining for 20min, opening the reaction kettle, and taking out a reaction product. Washing with anhydrous methanol for 3 times by centrifugation (8000rpm), vacuum drying, and storing at low temperature to obtain final product, i.e. star hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONOATE) with hollow polydopamine as core and 3 generation dendritic polyamide as arm and loaded with antibiotic Piperacillin (PRL) and Nitric Oxide (NO). The mass ratio of the h-PDA @ PAMAM/PRL to the sodium methoxide is 1: 10; the anhydrous methanol was calculated as 0.5g of sodium methoxide per 10 mL.
Example 10
Synthesis of final product h-PDA @ PAMAM/PRL/NONOate by co-loading antibiotic Piperacillin (PRL) and Nitric Oxide (NO) gas
(1) Dispersing the star-shaped hollow nano material (h-PDA @ PAMAM) obtained in the certain mass example 8 into pure water with a certain volume at the room temperature of 35 ℃, then slowly adding PRL with a certain mass, continuously stirring for 24h, finally centrifuging (5000rpm) and washing with pure water for 2 times, and finally obtaining the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/PRL). The mass ratio of the h-PDA @ PAMAM to the PRL is 1: 5; the pure water is calculated by adding 50mg of h-PDA @ PAMAM into every 10 mL;
(2) and (2) drying the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/PRL) obtained in the step (1), dispersing the dried star-shaped hollow nano material in absolute methanol, adding sodium methoxide, stirring for 10min, placing the mixture in a high-pressure reaction kettle, sealing and detecting the air tightness. Keeping the reaction kettle (10psi) for 15min by using high-purity nitrogen to remove air in the reaction kettle, and then introducing NO gas (40psi) to react for 3-7 days at room temperature. After the reaction is finished, the NO is discharged by high-purity nitrogen of 10psi and continuously maintained for 20min, then the reaction kettle is opened, and the reaction product is taken out. Washing with anhydrous methanol for 2 times by centrifugation (5000rpm), vacuum drying, and storing at low temperature to obtain final product, i.e. star hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONOATE) with hollow polydopamine as core and 3 generation dendritic polyamide as arm and loaded with antibiotic Piperacillin (PRL) and Nitric Oxide (NO). The mass ratio of the h-PDA @ PAMAM/PRL to the sodium methoxide is 1: 5; the anhydrous methanol was calculated by adding 0.1g of sodium methoxide per 10 mL.
Example 11
5mg of the hollow polydopamine nanoparticles (pDA HNP) obtained in example 1 are dispersed into 10mL of ethanol solution, and the solution is ultrasonically dispersed uniformly and then is dripped onto a copper net for observation, and the result is shown in figure 1, the particle size of the polydopamine nanoparticles (pDA HNP) is uniform, the hollow structure is obvious, and the fact that the hollow polydopamine nanoparticles (pDA HNP) are successfully prepared is proved.
Example 12
1mg of the hollow polydopamine nanoparticles (pDAHNP) obtained in example 2 and the azide group-modified hollow polydopamine nanoparticles (pDA-N) obtained in example 3 were each collected3HNP), the star-shaped hollow nano material (h-PDA @ PAMAM) taking hollow polydopamine as a core and dendritic polyamide as an arm obtained in example 7, the PRL-loaded star-shaped hollow nano material (h-PDA @ PAMAM/PRL) obtained in example 9, and the star-shaped hollow nano particles (h-PDA @ PAMAM/PRL/NONONOATE) taking hollow polydopamine as a core and 3 generations of dendritic polyamide as an arm co-loaded with antibiotics Piperacillin (PRL) and Nitric Oxide (NO) are dispersed in 1mL of pure water and then the surface potential changes of different nano particles are measured, and as shown in FIG. 2, we find that the nano particles subjected to azide modification have lower potential compared with the hollow polydopamine nano particles (pDAHNP) because of negative potential changes caused by successful modification of azide groups. However, the potential of the h-PDA @ PAMAM obtained by clicking the dendritic PAMAM to the hollow polydopamine nanoparticles is changed greatly, and the original negative potential is changed into a positive potential, which indicates that the PAMAM is successfully modified to the surfaces of the hollow polydopamine nanoparticles. After the PRL and the NO are continuously loaded to the h-PDA @ PAMAM, compared with the PRL, the potential of the surface of the nanoparticle after the NO is loaded is greatly reduced, because the NO is loaded to cause the occupation of secondary amine groups, the potential of the surface of the nanoparticle is obviously reduced, and the successful preparation of the final product, namely the star-shaped hollow nanoparticle h-PDA @ PAMAM/PRL/NONOATE, is proved.
Example 13
Infrared spectrum characterization is carried out on the hollow polydopamine nanoparticles (pDAHNP) obtained in example 1, the hollow polydopamine-core and dendritic polyamide-arm star-shaped hollow nanomaterials (h-PDA @ PAMAM) obtained in example 7, the single-load PRL star-shaped hollow nanomaterials (h-PDA @ PAMAM/PRL) obtained in example 9, and the double-antibiotic Piperacillin (PRL) and Nitric Oxide (NO) -loaded star-shaped hollow nanoparticles (h-PDA @ PAMAM/PRL/NONONAte) by a potassium bromide tabletting method. The results are shown in FIG. 3 for h-PDA @ PAMAM is at 3265cm-1Is the absorption peak of N-H; 2940cm-1And 2840cm-1 is-CH 2-and the absorption peak of the asymmetric stretching vibration and the symmetric stretching vibration; 1650cm-1And 1560cm-1The band is the characteristic band of amide group, which is called amide I band and amide II band, the former is caused by carboxyl stretching vibration, the latter is caused by N-H bond bending deformation vibration and C-N bond stretching vibration in-CONH-; 1120cm-1And 1035cm-1Respectively, the stretching vibration peak of primary amine and tertiary amine, and the two absorption peaks are weaker. The analysis result of the infrared spectrum shows that the sample contains characteristic groups such as-NH 2, -CH2-, -CONH-and the like, and the structural characteristics of the PAMAM are consistent, and the result proves that the surface of the hollow polydopamine nano particle (pDA HNP) successfully modifies the third generation of dendritic polyamide, namely the hollow polydopamine is used as a core and the dendritic polyamide is used as an arm star-shaped hollow nano material (h-PDA @ PAMAM). The same h-PDA @ PAMAM/PRL spectrum was found to be 3046cm-1、1607cm-1And 1500cm-1Is a characteristic absorption peak of anti-biological PRL lactam bond, 1374cm-1And 2970cm-1Is PCR upper-C (CH)3)2Characteristic absorption peaks, thus demonstrating successful loading of the antibiotic PRL. Also at 3265cm-1Decrease of characteristic peak at N-H, and 1300cm-1The NONONOate characteristic absorption peak appears, the NO is proved to be successfully modified and loaded, and the star-shaped hollow nano-particle (h-PDA @ PAMAM/PRL/NONONOate) loaded with the antibiotics Piperacillin (PRL) and Nitric Oxide (NO) in a double way is proved to be successfully prepared.
Example 14
10mg of final product h-PDA @ PAMAM/PRL/NONONAte co-loaded with the antibiotic Piperacillin (PRL) and Nitric Oxide (NO) gas obtained in example 9, and h-PDA @ PAMAM/NVA and h-PDA @ PAMAM/CFM obtained by single loading of other antibiotics (vancomycin (NVA) and Cefixime (CFM)) were dispersed in 10mL of PBS (pH7.4) buffer and placed in a constant temperature shaking table at 37 ℃, 1mL of solution was centrifuged after the nanoparticles were shaken uniformly at intervals, supernatant was collected, absorbance of the released solution of the nanoparticles h-PDA PAMAM/PRL/NONONONAte at OD540 nm was measured using Grids, and a NO standard curve (Y ═ 0.0052X-0.0118, R @ 0.20.999; wherein X isTable NO molarity, Y represents absorbance using OD 540) the NO release of the final h-PDA @ PAMAM/PRL/NONOate nanoparticles at each time point was calculated. Meanwhile, the absorbance of the supernatant at OD 206nm, 288nm and 281nm was measured using HPLC and the PRL, CFM and NVA release amounts were calculated. The experimental result is shown in fig. 4, we find that the release speed of NO is relatively high, the release half-life period is about 1.5h, and the release period is 20h, however, the release rate of PRL, CFM and NVA is relatively low, we speculate that the release rate of the small molecule drug is slow due to the internal structure of the dendritic PAMAM, and the small molecule drug has a certain slow release effect, and the potential advantages of the h-PDA @ PAMAM nanoparticle in the aspect of drug delivery are indirectly proved due to the loading characteristics of various antibiotics.
Example 15
Respectively taking the same mass of the star-shaped hollow nano material (h-PDA @ PAMAM) taking the hollow polydopamine as the core and the dendritic polyamide as the arm obtained in the example 8, the star-shaped hollow nano material (h-PDA @ PAMAM/PRL) taking the PRL loaded hollow polydopamine as the core and the 3 generations of dendritic polyamides as the final product, culturing the star-shaped hollow nano particles (h-PDA @ PAMAM/PRL/NOnoate) taking the hollow polydopamine as the core and the 3 generations of dendritic polyamides as the arms, and loading antibiotics Piperacillin (PRL) and Nitric Oxide (NO) together with escherichia coli with the same concentration and in the logarithmic phase for growth for 4h, centrifugally removing the material, plating the bacterial liquid and placing the bacterial liquid in an incubator for 12h, then calculating the bacterial number on the agar plate, and finding that the antibiotic-loaded h-PDA @ PAM/PRL is obviously enhanced in antibacterial effect compared with the h-PDA @ PAMAM as shown in figure 5, however, the sterilization effect is still not obvious, but the h-PDA @ PAMAM/PRL/NONOATE sterilization effect after NO loading is obviously improved, which shows that compared with a single antibacterial mode, the synergistic effect of the NO and the PRL acting on bacteria together is obvious, and the rationality of the material design is also intuitively proved.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. A preparation method of a star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis is characterized by comprising the following steps:
(1) synthesis of hollow polydopamine nanoparticles
Preparing silicon dioxide nanoparticles, dispersing the silicon dioxide nanoparticles in a buffer solution, adding dopamine hydrochloride, and stirring to react for 24-48 hours to prepare poly-dopamine-coated silicon dioxide nanoparticles; dispersing the poly-dopamine-coated silicon dioxide nanoparticles into a mixed solution of hydrofluoric acid and ammonium fluoride, and continuing to react for 24-48 h to prepare hollow poly-dopamine nanoparticles;
(2) hollow poly-dopamine nanoparticle modified by azide group
Dissolving azidoacetic acid in an organic solvent, and then adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide for activation for 30 min-4 h; dispersing the hollow polydopamine nanoparticles prepared in the step (1) in pure water, then adding the dispersed hollow polydopamine nanoparticles into an azidoacetic acid mixed solution after activation, and reacting at room temperature for 12-24 hours to prepare azido group modified hollow polydopamine nanoparticles;
(3) synthesis of star-shaped hollow nano material with hollow polydopamine as core and dendritic polyamide as arm
Dispersing the azide group modified hollow polydopamine nano particles prepared in the step (2) in an organic solvent, introducing protective gas, adding a dendritic polyamide aqueous solution, then adding copper sulfate pentahydrate and sodium ascorbate, continuing introducing the protective gas, and reacting at 40-75 ℃ for 24-48 h to prepare the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis.
2. The preparation method of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial according to claim 1, characterized in that in the step (1):
the silica nanoparticles are prepared by the following steps: uniformly mixing 25% ammonia water, absolute ethyl alcohol and pure water, adding 99% tetraethoxysilane to change the system solution from transparent to white, continuously stirring for 5-10 h, centrifuging to remove the suspension, and alternately washing with ethanol and pure water to obtain silicon dioxide nanoparticles; the volume ratio of the ammonia water to the absolute ethyl alcohol to the pure water to the ethyl orthosilicate is 1: 10-30: 2-5: 0.5 to 2;
the buffer solution is a tris buffer solution with pH of 7.5-9;
the mass ratio of the silicon dioxide nanoparticles to the dopamine hydrochloride is 1: 1-5;
the mass volume ratio of the poly-dopamine-coated silicon dioxide nanoparticles to the mixed solution of hydrofluoric acid and ammonium fluoride is 50-100 mg: 5-10 mL;
the molar concentration ratio of hydrofluoric acid to ammonium fluoride in the mixed solution of hydrofluoric acid and ammonium fluoride is 1:2 to 5.
3. The preparation method of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial according to claim 1, characterized in that in the step (2):
the mole ratio of the hollow polydopamine nanoparticles to the 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride to the N-hydroxysuccinimide to the azidoacetic acid is (1: 1) - (10): 1-10: 1-10;
the organic solvent is at least one of N-N-dimethylformamide, dimethyl sulfoxide and methanol; adding 1-5 g of azido acetic acid into each 10mL of organic solvent;
the pure water is calculated by adding 0.1-5 g of azido acetic acid into every 10 mL.
4. The preparation method of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial according to claim 1, characterized in that in the step (3):
the mass ratio of the hollow poly-dopamine nano particles modified by the azide groups to the dendritic polyamide is 1: 50-100;
the molar ratio of the dendritic polyamide to the copper sulfate pentahydrate to the sodium ascorbate is 1: 7-14: 1-3: 3-9;
the organic solvent is at least one of dimethyl sulfoxide, DMF and tetrahydrofuran, and is calculated by adding 50-100 mg of azide group modified hollow polydopamine nano particles into every 10 mL; the amount of the pure water is calculated by adding 1-3 g of dendritic polyamide into every 10 mL;
and the protective gas is nitrogen, and the dendritic polyamide aqueous solution is added after the protective gas is introduced for 10-30 min.
5. A star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis is characterized in that the star-shaped hollow nano material is prepared by using hollow polydopamine as a core and dendritic polyamide as an arm through the preparation method of any one of claims 1 to 4.
6. The application of the star-shaped hollow nanometer material capable of simultaneously loading NO and antibiotics for synergistic antibiosis as claimed in claim 5 is characterized in that the star-shaped hollow nanometer material is used for preparing an antibacterial drug loading nitric oxide and antibiotics.
7. The use according to claim 6, wherein the antibiotic is piperacillin, vancomycin, cefixime, ampicillin, tetracycline or norfloxacin.
8. The star-shaped nanometer material capable of releasing NO and antibiotics for synergistic antibiosis is characterized by comprising the following specific preparation steps:
a. dispersing the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis, which is disclosed by claim 5, in pure water at room temperature, then adding a certain mass of antibiotics, and continuously stirring for 24-48 h to obtain the star-shaped hollow nano material loaded with the antibiotics;
b. drying the antibiotic-loaded star-shaped hollow nano material, dispersing the dried star-shaped hollow nano material in anhydrous methanol, tetrahydrofuran or acetonitrile, adding sodium methoxide, stirring for 10-30 min, placing the mixture in a high-pressure reaction kettle, sealing the high-pressure reaction kettle, introducing high-purity nitrogen to remove air in the high-pressure reaction kettle, introducing NO gas, and reacting at room temperature for 3-7 days to prepare the NO and antibiotic-loaded star-shaped nano material for synergistic antibiosis.
9. Star-shaped nanomaterial capable of releasing NO and antibiotics for synergistic antibacterial use according to claim 8,
the mass ratio of the star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibiosis to the antibiotics is 1: 1-10;
the mass ratio of the antibiotic-loaded star-shaped hollow nano material to sodium methoxide is 1: 5-20;
introducing high-purity nitrogen to maintain the reaction kettle at 10-20 psi for 5-15 min, and removing air in the reaction kettle; the pressure of the high-purity nitrogen is 10-20 psi, and the pressure of the NO gas is 40-80 psi.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911016279.1A CN110787298B (en) | 2019-10-24 | 2019-10-24 | Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911016279.1A CN110787298B (en) | 2019-10-24 | 2019-10-24 | Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110787298A CN110787298A (en) | 2020-02-14 |
| CN110787298B true CN110787298B (en) | 2021-09-17 |
Family
ID=69441262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911016279.1A Active CN110787298B (en) | 2019-10-24 | 2019-10-24 | Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110787298B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111588860B (en) * | 2020-05-15 | 2023-04-07 | 暨南大学 | Spherical PAMAM polymer capable of simultaneously loading NO and amphotericin B as well as preparation method and application thereof |
| CN111729131A (en) * | 2020-08-07 | 2020-10-02 | 北京邦塞科技有限公司 | Intelligent controlled-release medicine-carrying bone cement and preparation method and application thereof |
| CN114887057B (en) * | 2022-05-07 | 2023-05-09 | 北京化工大学 | Yolk-Shell type nanoparticle as well as preparation method and application thereof |
| CN114949253B (en) * | 2022-06-01 | 2022-12-16 | 暨南大学附属第一医院（广州华侨医院） | Double-drug-loading polyrotaxane nano drug delivery system and preparation method and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199297A (en) * | 2011-03-25 | 2011-09-28 | 中山大学孙逸仙纪念医院 | Star-shaped cationic polymer, preparation method thereof and application thereof |
| CN106046382A (en) * | 2016-05-25 | 2016-10-26 | 暨南大学 | Nitric oxide loaded cationic polymer, preparation method therefor and application of nitric oxide loaded cationic polymer |
| CN107661504A (en) * | 2017-10-25 | 2018-02-06 | 暨南大学 | A kind of golden nanometer particle of dendritic macromole modification and its preparation method and application |
| CN107739448A (en) * | 2017-10-25 | 2018-02-27 | 暨南大学 | Magnetic control discharges nitric oxide production composite film material and its preparation method and application |
| CN107739506A (en) * | 2017-10-25 | 2018-02-27 | 暨南大学 | Light-operated nitric oxide production composite film material of release and its preparation method and application |
| CN107812188A (en) * | 2017-10-25 | 2018-03-20 | 暨南大学 | Magnetic material with nitric oxide/photo-thermal synergetic antibacterial effect and preparation method and application |
| CN108969769A (en) * | 2018-07-11 | 2018-12-11 | 暨南大学 | Penicillin and nitric oxide production polymer and its preparation method and application are loaded simultaneously |
-
2019
- 2019-10-24 CN CN201911016279.1A patent/CN110787298B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199297A (en) * | 2011-03-25 | 2011-09-28 | 中山大学孙逸仙纪念医院 | Star-shaped cationic polymer, preparation method thereof and application thereof |
| CN106046382A (en) * | 2016-05-25 | 2016-10-26 | 暨南大学 | Nitric oxide loaded cationic polymer, preparation method therefor and application of nitric oxide loaded cationic polymer |
| CN107661504A (en) * | 2017-10-25 | 2018-02-06 | 暨南大学 | A kind of golden nanometer particle of dendritic macromole modification and its preparation method and application |
| CN107739448A (en) * | 2017-10-25 | 2018-02-27 | 暨南大学 | Magnetic control discharges nitric oxide production composite film material and its preparation method and application |
| CN107739506A (en) * | 2017-10-25 | 2018-02-27 | 暨南大学 | Light-operated nitric oxide production composite film material of release and its preparation method and application |
| CN107812188A (en) * | 2017-10-25 | 2018-03-20 | 暨南大学 | Magnetic material with nitric oxide/photo-thermal synergetic antibacterial effect and preparation method and application |
| CN108969769A (en) * | 2018-07-11 | 2018-12-11 | 暨南大学 | Penicillin and nitric oxide production polymer and its preparation method and application are loaded simultaneously |
Non-Patent Citations (2)
| Title |
|---|
| Dendritic Fe3O4@Poly(dopamine)@PAMAM Nanocomposite as Controllable NO-Releasing Material: A Synergistic Photothermal and NO Antibacterial Study;Siming Yu等;《Adv. Funct. Mater》;20181231;第1-14页 * |
| Polydopamine Hollow Nanoparticle Functionalized with N-diazeniumdiolates as a Nitric Oxide Delivery Carrier for Antibacterial Therapy;Dongsik Park等;《Adv. Healthcare Mater》;20161231;第2019-2024页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110787298A (en) | 2020-02-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110787298B (en) | Preparation and application of star-shaped hollow nano material capable of simultaneously loading NO and antibiotics for synergistic antibacterial | |
| Lu et al. | Synergistic bactericidal activity of chlorhexidine-loaded, silver-decorated mesoporous silica nanoparticles | |
| JP6266510B2 (en) | Metal chelate compositions and methods for controlling the growth or activity of living cells or organisms | |
| Upadya et al. | Role of efflux pump inhibitors on the antibiofilm efficacy of calcium hydroxide, chitosan nanoparticles, and light-activated disinfection | |
| CN106637992A (en) | Photodynamic antibacterial cellulose material and preparation method thereof | |
| JP2016529325A (en) | Antimicrobial compounds and methods for their production and use | |
| CN110123801B (en) | Application of multi-arm AIE molecule in preparation of antibacterial drug and antibacterial drug | |
| JP2008528761A (en) | Β-cyclodextrin derivatives as antibacterial agents | |
| CN108969769B (en) | Polymer simultaneously carrying penicillin and nitric oxide as well as preparation method and application thereof | |
| US9308185B2 (en) | Glyco-substituted dihydroxy-chlorins and β-functionalized chlorins for anti-microbial photodynamic therapy | |
| Zhao et al. | Antibacterial carbon dots: mechanisms, design, and applications | |
| Bajaj et al. | Stabilized cationic dipeptide capped gold/silver nanohybrids: Towards enhanced antibacterial and antifungal efficacy | |
| TW201811180A (en) | Polyamine carbon dot and use thereof | |
| CN107513145A (en) | The synthesis of biodegradable amphiphilic block class antibacterial peptide and the preparation method and application of class antibacterial peptide vesica | |
| Gavara et al. | Broad-spectrum photo-antimicrobial polymers based on cationic polystyrene and rose bengal | |
| CN107596452B (en) | Preparation method and application of intelligent antibacterial coating sensitive to bacteria | |
| US20230082968A1 (en) | Exogenous nitric oxide for improved susceptibility and lowered antibiotic resistance in resistant respiratory bacteria | |
| Shaikh et al. | Nanobiotechnological approaches against multidrug resistant bacterial pathogens: An update | |
| JP4970931B2 (en) | Diamine-based guanidine derivatives with antibacterial activity | |
| CN105555312A (en) | Medicinal composition for promoting synthesis of protoporphyrin IX | |
| CN115433367B (en) | Porphyrin COF material, preparation method and application | |
| US10800732B2 (en) | Substituted malonamides and their use as antibacterial drugs | |
| CN114272246A (en) | Application of uracil in preparing anti-infective medicine | |
| WO2018010403A1 (en) | Use of medicinal nanomaterial composition dg-5 applied to anti-drug resistant bacteria | |
| AU2018272839A2 (en) | Fractionated antimicrobial compositions and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |