CN110559451B - Nano medicine and its preparing method and use - Google Patents
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- CN110559451B CN110559451B CN201910792487.4A CN201910792487A CN110559451B CN 110559451 B CN110559451 B CN 110559451B CN 201910792487 A CN201910792487 A CN 201910792487A CN 110559451 B CN110559451 B CN 110559451B
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
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- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
- A61K49/0089—Particulate, powder, adsorbate, bead, sphere
- A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
- A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
Abstract
The invention relates to a nano-drug and a preparation method and application thereof, wherein the nano-drug comprises an amphiphilic cyanine dye and a drug for treating gram-positive bacteria; the preparation method comprises the following steps: and adding the amphiphilic cyanine dye and a medicine for treating gram-positive bacteria into a buffer solution to obtain the nano-medicine. The amphiphilic cyanine dye in the nano-drug provided by the invention can form stable vesicles to entrap drugs for treating gram-positive bacteria, the aggregates are dispersed after the infected parts of the gram-positive bacteria are enriched in a targeted manner, so that the fluorescence signals are enhanced, the infected parts of the bacteria can be effectively marked and effectively treated, and the novel material which combines diagnosis and treatment integrated functions and is developed based on the nano-technology has wide application prospects clinically.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a nano-medicine and a preparation method and application thereof.
Background
The diseases caused by bacterial infection are diseases with high morbidity and mortality. In clinical treatment, doctors' treatment of infection is mainly based on hemogram examination, and the data provided by the single detection means is not enough to guide doctors to make accurate judgment. Therefore, there is an urgent need to develop a quantitative diagnosis method for in vivo infection while image-guiding the doctor's medication to solve the above-mentioned troublesome problems. At present, the non-invasive detection means studied is an isotope in-situ injection labeling technology, but the specificity of the label is poor, and the detection resolution and the detection depth are limited, so that the wide application in clinic is difficult.
For the treatment of bacterial infections, antibiotics have gradually been widely used in the medical field since the discovery of penicillin, opening up a new effective way for the prevention and treatment of diseases. However, one of the biggest challenges facing modern medicine is increased bacterial resistance due to uncontrolled use of antibiotics. The emergence of antibiotic-resistant bacteria has limited the clinical use of antibiotics, including bacterial infections caused by bacteria, viruses, fungi, and the like in chemotherapy, transplantation, and surgery. One of the reasons for the use of a large number of antibiotics is that high-concentration enrichment of drugs cannot be achieved at the site of bacterial infection, so that increasing the local drug concentration at the infected site is an important way to reduce the dosage of the drugs, and the drug targeting at the site of bacterial infection is released by using a carrier material as one of the ways to increase the local drug concentration. The carrier materials for carrying drugs reported at present are polymer materials, liposomes and the like.
CN107970226A discloses a nano-carrier drug, a preparation method and an application thereof, wherein the nano-carrier drug takes polylactic acid-glycolic acid as a carrier, NVP-BEZ235 and chlorin E6 are loaded, and the surface of the carrier is subjected to PEGylation by lecithin and distearoylphosphatidylethanolamine-polyethylene glycol. By using polylactic acid-glycolic acid as a carrier and combining chlorin E6 and NVP-BEZ235 as a therapeutic agent, the combination of photodynamic therapy and biotherapy can be realized, and the killing effect on tumor cells can be improved by synergistic effect. The surface of the nano-particles is subjected to PEGylation by lecithin and DSPE-PEG, so that the blood circulation time can be prolonged, the removal of reticuloendothelial tissues is reduced, the treatment effect is further ensured, the treatment effect of tumor treatment, particularly triple negative breast cancer, is remarkable, and the application prospect is wide.
CN105708799A discloses a nanostructured lipid carrier pharmaceutical composition and a preparation method thereof, wherein the pharmaceutical composition comprises the following components by weight percent: 0.02% -1% of insoluble drug, solid lipid material: 0.3 to 15 percent of water-soluble emulsifier, 0.5 to 20 percent of liquid lipid material, 0.8 to 10 percent of fat-soluble emulsifier, 1 to 15 percent of water-soluble emulsifier and 39 to 97.38 percent of aqueous solvent. The nano-structure lipid carrier prepared by the invention greatly overcomes the defects that insoluble drugs are not easy to dissolve in water, the oral bioavailability is low and the like, and provides an alternative drug delivery system for the insoluble drugs.
CN101862457A discloses a macromolecular carrier drug and a preparation method and application thereof. The high molecular carrier drug is obtained by photo-immobilizing TNF-alpha and IFN-gamma on a degradable biological material and carrying out surface modification on the degradable biological material. The base material used by the high molecular carrier drug is bioplastic, has no toxicity or irritation, good biocompatibility, biodegradability and absorption, high strength, good plasticity and easy processing and forming, can make the drug effects of TNF-alpha and IFN-gamma more durable and effective after being prepared into the drug, has good treatment effect and small toxic and side effect when being used for preparing the drug for treating ovarian cancer, and is suitable for wide application in medical treatment.
The three patents disclose that several typical carrier drugs can only treat diseases, and the effect is single.
Therefore, there is a need in the art to develop a novel nano-drug that not only can treat diseases, but also can quantitatively detect the content of pathogenic cells.
Disclosure of Invention
In view of the shortcomings of the prior art, one of the objectives of the present invention is to provide a nano-drug. The nano-drug can be used for specific detection and treatment of gram-positive bacteria.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a nano-drug, which comprises an amphiphilic cyanine dye and a drug for treating gram-positive bacteria.
The nano-drug provided by the invention is a near-infrared nano-drug, the amphiphilic cyanine dye can form a stable vesicle which can emit a fluorescence signal under the irradiation of near-infrared light and has the characteristics of ultraviolet absorption blue shift and fluorescence quenching, the vesicle entraps the drug for treating gram-positive bacteria, the fluorescence signal is enhanced due to the dissolution of aggregates after the infection part of the gram-positive bacteria is targeted and enriched, the bacterial infection part can be effectively marked and can be effectively treated; the kit has unique specificity, can effectively distinguish gram-positive bacterial infection, negative bacterial infection and nonbacterial infection inflammation, realizes quantitative detection of the gram-positive bacterial infection in vivo by combining the linear relation of a fluorescence signal and the bacterial concentration, images the bacterial infection part and guides the medication.
The nano-drug has two functions of treatment and detection, and the new material developed by combining the diagnosis and treatment integrated function and the nano-technology has wide application prospect in clinic.
Preferably, the molar weight ratio of the amphiphilic cyanine dye to the drug for treating gram-positive bacteria is 3:1-20:1, such as 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, etc., preferably 10: 1.
Preferably, the amphiphilic cyanine dye comprises any one or at least two combinations of compounds shown in formulas I to VIII;
the R is1And R3All are hydrophobic groups, and the hydrophobic groups contain C1-C11 alkyl and/or C6-C24 aryl;
the R is2Selected from F, Cl, Br or I;
said X—Is selected from Br—Or I—；
Said n is 0, 1 or 2, preferably 1.
The invention preferably selects the amphiphilic cyanine dyes with the specific structures by changing the length of the hydrophobic carbon chain or the number of benzene rings (R) of the cyanine dyes1And R3) To modulate the amphiphilicity of the cyanine dyes.
The synthesis methods of the compounds represented by formulas I to VIII are prior art, and refer to patents CN109796779A and CN109054428A, and those skilled in the art can select specific synthesis methods according to common general knowledge and actual needs.
Preferably, said R is1And R3Each independently selected from any one of the following groups:
m is an integer of 1 to 11, such as 2,3, 4, 5, 6, 7, 8, 9, 10, etc.;
s is an integer from 2 to 4, for example 3;
in the present invention,
represents an access bond to a group.
Preferably, the amphiphilic cyanine dye comprises a compound represented by formula I.
Preferably, the amphiphilic cyanine dye includes the following compounds:
said X—Selected from Cl—、Br—Or I—；
The R is2Selected from F, Cl, Br or I, preferably Cl.
The above compounds are preferred in the present invention because nanovesicles formed from these compounds can specifically recognize gram-positive bacteria with higher accuracy, and can specifically dissolve and turn on fluorescence signals.
Preferably, the medicine for treating gram-positive bacteria comprises amino acid medicines and/or glycopeptide medicines, and preferably glycopeptide medicines.
Preferably, the amino acid based drug comprises HNP-1, PR-39, LL-37, HBD-3, heliomycin (heliomycin), alpha-defensin 6 andA9k (prepared by the method described in biomatic macromolecules 2010; 11(2): 402-.
The amino acid sequences of the amino acid medicaments are as follows:
HNP-1：ACYCRIPACIAGERRYGTCIYQGRLWAFCC；
PR-39：RRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP；
LL-37：LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES；
HBD-3：GIINTLQKYYCRVRGGRCAVLSCLPKEEQIGKCSTRGRKCCRRK；
Indolicidin：ILPWKWPWWPWRR-NH2；
heliomycin:DKLIGSCVWGAVNYTSDCNGECKRRGYKGGYCGSFANVNC WCET；
α-defensin 6：AFTCHCRRSCYSTEYSYGTCTVMGINHRFCCL；
A9K：AAAAAAAAAK。
except for A9In addition to K, the above amino acid drugs are commercially available.
Preferably, the glycopeptide drug comprises any one or combination of at least two of vancomycin derivatives, norvancomycin derivatives, teicoplanin derivatives, leiomycins, ramoplanin, oritavancin and daptomycin, and oritavancin is preferred.
The glycopeptide drugs can be prepared by methods in the prior art or can be obtained by commercial methods.
Preferably, the amphiphilic cyanine dye is in a nanovesicle form, and the drug for treating gram-positive bacteria is encapsulated in the nanovesicle.
Preferably, the nanovesicle has the characteristics of ultraviolet absorption blue shift and fluorescence quenching.
Preferably, the dosage form of the nano-drug comprises a solution or a dry powder.
Preferably, the route of administration of the nano-drug comprises intravenous injection or topical spray.
The second purpose of the present invention is to provide a method for preparing the nano-drug, which comprises the following steps: and adding the amphiphilic cyanine dye and a medicine for treating gram-positive bacteria into a buffer solution to obtain the nano-medicine.
Preferably, the molar weight ratio of the amphiphilic cyanine dye to the drug for treating gram-positive bacteria in the buffer solution is 3:1-20:1, preferably 10: 1.
Preferably, the amphiphilic cyanine dye and the drug for treating gram-positive bacteria are added by a syringe pump.
Preferably, the amphiphilic cyanine dye and the drug for treating gram-positive bacteria are added to the buffer solution at the same time.
Preferably, the amphiphilic cyanine dye and the gram-positive bacterium treatment drug are added to the buffer solution while stirring.
Preferably, the buffer solution includes any one or a combination of at least two of pure water, a phosphate buffer solution, a Tris-HCl buffer solution, and a HEPES buffer solution.
Preferably, after the amphiphilic cyanine dye and the gram-positive bacterium-treating drug are added to the buffer solution, the free gram-positive bacterium-treating drug is removed by dialysis.
The invention also aims to provide application of the nano-medicament in preparing a material or medicament for treating gram-positive bacteria.
Compared with the prior art, the invention has the following beneficial effects:
the nano-drug provided by the invention is a near-infrared nano-drug, the amphiphilic cyanine dye can form a stable vesicle which can emit a fluorescence signal under the irradiation of near-infrared light and has the characteristics of ultraviolet absorption blue shift and fluorescence quenching, the vesicle entraps the drug for treating gram-positive bacteria, targets and enriches infected parts of the gram-positive bacteria, aggregates are dispersed after being triggered by the bacterial wall specificity of the gram-positive bacteria, so that the fluorescence signal is enhanced, the infected parts of the bacteria can be effectively marked and can be effectively treated; the kit has unique specificity, can effectively distinguish gram-positive bacterial infection, negative bacterial infection and nonbacterial infection inflammation, realizes quantitative detection of the gram-positive bacterial infection in vivo by combining the linear relation of a fluorescence signal and the bacterial concentration, images the bacterial infection part and guides the medication.
The nano-drug has two functions of treatment and detection, and the new material developed by combining the diagnosis and treatment integrated function and the nano-technology has wide application prospect in clinic.
Drawings
FIG. 1 is a MALDI-TOF spectrum of the nanomolecular amphiphilic cyanine dye Cy obtained in Synthesis example 1.
Fig. 2 is a TEM image of the nano-drug a obtained in example 1.
FIG. 3 is an image showing the specificity of the nano-drug A for different bacteria in test example 1 (wherein, a-Staphylococcus aureus, b-Staphylococcus epidermidis, c-Escherichia coli, d-Serratia, e-Pseudomonas aeruginosa).
FIG. 4 is a graph of near-infrared fluorescence imaging of the nano-drug A in test example 2 for detection of different bacteria (wherein, a-Staphylococcus aureus, b-Staphylococcus epidermidis, c-Escherichia coli, d-Serratia, e-Pseudomonas aeruginosa).
FIG. 5 is a graph of near infrared fluorescence imaging of various treatment conditions in test example 3 (where a-blank control, b-has been injected with oritavancin, and c-has been injected with Nanrug A).
FIG. 6 is a diagram of quantitative analysis of near-infrared fluorescence imaging of mice in test example 3 after treatment with various drugs.
Fig. 7 is a graph of the ultraviolet absorption spectrum of amphiphilic cyanine dye Cy monomers and aggregates.
Fig. 8 is a graph of fluorescence spectra of amphiphilic cyanine dye Cy monomers and aggregates.
Detailed Description
For the purpose of facilitating an understanding of the present invention, the present invention will now be described by way of examples. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
Synthesis example 1:
the synthesis example provides a preparation method of an amphiphilic cyanine dye Cy, which specifically comprises the following steps:
2,3, 3-trimethylindole (20 mmol) and bromododecane (20 mmol) were refluxed in a toluene solution for 24 hours. The reaction was cooled to room temperature, then filtered and washed 3 times with petroleum ether (10 ml). The product 1 was obtained as a violet solid and used in the next reaction without further purification.
Dimethylformamide (40 ml) and dichloromethane (40 ml) were mixed in an ice bath, and then a mixed solution of phosphorus oxychloride (19 ml) and dichloromethane (19 ml) was added dropwise. After the addition was complete, 5 g of cyclohexanone were added. The reaction solution is refluxed for 3 hours, and after the reaction is finished, the reaction solution is cooled to room temperature. The reaction was then added dropwise to a 100 g ice-water bath. The solution was left to stand at 4 ℃ overnight. Finally, filtration gave product 2 as a yellow solid. It was used in the next reaction without further isolation.
The above two reaction products 1(1 mmol), 2(1 mmol) and 41 mg of sodium acetate were dissolved in the acetic anhydride solution and stirred at room temperature for 3 hours. And separating and purifying a crude product obtained by the reaction by column chromatography to obtain a compound Cy.
MALDI-TOF characterization was performed on the amphiphilic cyanine dye Cy, and the results are shown in FIG. 1.
Example 1
The embodiment provides a preparation method of a nano-drug, which comprises the following specific steps:
by means of a syringe pump, 0.4. mu.g of oritavancin and 18. mu.g of the amphiphilic cyanine dye Cy were simultaneously and slowly injected into pure water at a concentration ratio of 1:10 (molar ratio) while stirring. Then, free drug molecules are removed by a dialysis method to obtain a nano drug a, wherein the amphiphilic cyanine dye Cy is in a nano vesicle form, oritavancin is wrapped in the amphiphilic cyanine dye Cy, and the form of the obtained nano particle is shown in fig. 2 through a Transmission Electron Microscope (TEM) image.
The amphiphilic cyanine dye Cy (monomer) in synthesis example 1 and the amphiphilic cyanine dye Cy nano-vesicle (aggregate) obtained in example 1 were tested by ultraviolet absorption spectroscopy (UV-2600, shimadzu) and fluorescence spectroscopy (F-280, tianjin hongkongdong science and technology ltd.), and ultraviolet absorption spectrograms of the amphiphilic cyanine dye Cy monomer and aggregate shown in fig. 7 were obtained, which show that the positions of absorption peaks of the monomer and aggregate are different, and fluorescence spectrograms of the amphiphilic cyanine dye Cy monomer and aggregate shown in fig. 8 were obtained, which show that the monomer has fluorescence and the aggregate is not fluorescent, and when the aggregate reaches a target position, the aggregate is decomposed into the monomer and has a fluorescent signal, thereby effectively labeling a bacterial infection site.
Test example 1 Nanoparticulate in vitro specific bacterial Targeted Observation
Selecting Staphylococcus aureus and Staphylococcus epidermidis as gram-positive bacteria model strains, Escherichia coli, Serratia and Pseudomonas aeruginosa as gram-negative bacteria model strains, and adding 1 × 10 of Bacillus subtilis8The bacterial culture solution of CFU and 40 mu mol/L nano-drug A are incubated for 30 minutes in the same volume, and the mixture is processed by a laser confocal microscope (resonant demo, lambda)EX650nm) and the results are shown in figure 3, wherein the bacteria of a, b, c, d and e in the figure are respectively staphylococcus aureus, staphylococcus epidermidis, escherichia coli, serratia and pseudomonas aeruginosa, and from figure 3, the bacteria of a and b are marked (the small circle point with the contrast different from the black background is a fluorescent signal), and c, d and e are completely in a dark state, so that the nano vesicle in the nano-drug A can specifically target the gram-positive bacteria, namely effectively distinguish gram-positive bacterial infection from gram-negative bacterial infection.
The bactericidal effect is determined by the Minimum Inhibitory Concentration (MIC) value, wherein the specific calculation method of the MIC is as follows:
preparation of MIC plate: aseptic operation, adding the antibacterial solution with different concentrations after dilution by multiple times into a sterilized 96-hole polystyrene plate, adding the liquid medicine into the 1 st to 11 th holes with 10 mu L of each hole, using no medicine in the 12 th hole as a growth control, freeze-drying, sealing and storing below-20 ℃ for later use.
Preparation of inoculum: diluting the bacterial suspension prepared by a growth method or a direct bacterial suspension method and having the concentration equivalent to 0.5 McLeod's turbidimetry standard by MH broth 1:1000, adding 100 mu L of the bacterial suspension into each hole, sealing the hole, putting the hole in a common air incubator at 35 ℃, and incubating for 16-24 h to judge the result. At this time, the drug concentrations in the 1 st to 11 th wells were 12.8, 6.4, 3.2, 1.6, 0.8, 0.4, 0.2, 0.1, 0.05, 0.025, 0.0125. mu.g/ml, respectively.
And (5) judging a result: the MIC was taken as the lowest drug concentration that completely inhibited bacterial growth in the wells. The MIC value calculated by the above method was 0.8 micrograms per ml (methicillin-resistant staphylococcus aureus). The result shows that the nano-drug A has good bactericidal effect.
Test example 2 detection of bacterial infection in vivo with Nanopaharmaceutical
Selecting 5 BALB/c mice with 6-8 weeks, and injecting 1 x 10 mice into the hind leg muscle respectively8And (3) culturing the bacteria culture solution of the CFU staphylococcus aureus, the staphylococcus epidermidis, the escherichia coli, the serratia and the pseudomonas aeruginosa for 48 hours to form five groups of mouse hind leg myositis models. 200 mu L of nano-drug A with the concentration of 200 mu M is respectively injected through tail veins, time nodes of 1 hour, 4 hours, 8 hours, 12 hours and 24 hours are selected, and imaging of the myositis infection at different time points is observed through a small animal optical 3D living body imaging system (IVIS spectrum, Perkin Elmer company).
Intercepting the imaging result of 1 hour, as shown in fig. 4, the bacteria of a, b, c, d and e in the figure are staphylococcus aureus, staphylococcus epidermidis, escherichia coli, serratia and pseudomonas aeruginosa respectively, the position pointed by the arrow in the figure is a bacterial infection part, the observation shows that the infection parts in a and b have fluorescence signals (in a dotted line), and no fluorescence signal is shown in c, d and e, which proves that the nano-drug A can selectively detect gram-positive bacteria and effectively distinguish gram-positive bacterial infection and gram-negative bacterial infection.
Furthermore, the gram-positive bacteria in the mouse can be quantitatively analyzed by the following specific method:
the fluorescence signal intensity is collected by a small animal fluorescence living body imaging system, and the bacterial concentration is calculated through the linear relation between the fluorescence signal and the bacterial concentration.
Wherein the linear relation standard curve of the fluorescence signal and the bacterial concentration is as follows:
y＝4.3×10-2x+7.47×106。
test example 3 treatment of Nanoparticulate in vivo bacterial infection
Selecting 3 BALB/c mice with 6-8 weeks, and injecting 1 x 10 in the hind leg muscle8And (3) forming a mouse hind leg myositis model by using the CFU methicillin-resistant staphylococcus aureus culture solution for 48 hours. 2 mice are selected, 200 muL of nano-drug A with the concentration of 200 muM and 200 muL of oritavancin with the concentration of 200 muM are respectively injected into the mice through tail veins, time nodes of 1 hour, 4 hours, 8 hours, 12 hours and 24 hours are selected, and the treatment effect of the mice on the myositis at different time points is observed through a small animal fluorescence living body imaging system instrument.
The time is prolonged to 4 days, the treatment conditions of three mice are recorded, and the results are shown in fig. 5, wherein the mice in a are not injected with the medicament (blank control group), the mice in b are injected with oritavancin, the mice in c are injected with the nano medicament A, the area in the dotted line is an infected part, and the infected part of the mice injected with the nano medicament A has a smaller fluorescence signal area (c), which shows that the bacteria content is greatly reduced after treatment and the curative effect is good; and compared with untreated mice, the fluorescence signal area of the infected part of the mice injected with oritavancin is only reduced in a small range, and the curative effect is poor.
The mean fluorescence signal value per unit area was obtained by quantitative analysis of the fluorescence imaging area of the infected sites, and figure 6, which shows that the mice in group a of the nano-drugs had the weakest fluorescence signal after treatment, indicating the best treatment effect.
The results of the test examples prove that the nano-drug provided by the invention can be used for specific detection and treatment of gram-positive bacteria, can specifically distinguish gram-positive bacteria infection and gram-negative bacteria infection, and has good curative effect.
Comparative example 1
The difference from example 1 is that the amphiphilic cyanine dye Cy is replaced by rhodamine of equal mass to obtain the nano-drug B.
Comparative test example 1
The difference from the test example 2 is that the nano-drug A is replaced by the nano-drug B, and the test result shows that the effect of the nano-drug A and the gram-positive bacteria can not generate fluorescence, namely the nano-drug B can not realize the specific marking of the gram-positive bacteria.
The applicant states that the present invention is illustrated by the above examples to show the detailed process equipment and process flow of the present invention, but the present invention is not limited to the above detailed process equipment and process flow, i.e. it does not mean that the present invention must rely on the above detailed process equipment and process flow to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (20)
1. A nano-drug, characterized in that the nano-drug comprises an amphiphilic cyanine dye and a drug for treating gram-positive bacteria; the amphiphilic cyanine dye is in a nano vesicle form, and the drug for treating gram-positive bacteria is wrapped in the nano vesicle;
the medicine for treating gram-positive bacteria comprises amino acid medicines and/or glycopeptide medicines;
the amphiphilic cyanine dye comprises a compound shown as a formula I;
the R is1And R3Each independently selected from any one of the following groups:
m is an integer of 1-11;s is an integer of 2-4; wherein the content of the first and second substances,
an access bond representing a group;
the R is2Selected from F, Cl, Br or I;
said X—Selected from Cl—、Br—Or I—；
And n is 1.
2. The nano-drug of claim 1, wherein the molar weight ratio of the amphiphilic cyanine dye to the drug for treating gram-positive bacteria is 3:1-20: 1.
3. The nano-drug of claim 2, wherein the molar weight ratio of the amphiphilic cyanine dye to the drug for treating gram-positive bacteria is 10: 1.
4. The nano-drug of claim 1, wherein the amphiphilic cyanine dye comprises the following compounds:
said X—Selected from Cl—、Br—Or I—；
The R is2Selected from F, Cl, Br or I.
5. The nano-drug of claim 4, wherein X is—Is Cl— 。
6. The nano-drug of claim 1, wherein the drug for treating gram-positive bacteria is a glycopeptide drug.
7. According to claim1, the nano-drug is characterized in that the amino acid drugs comprise HNP-1, PR-39, LL-37, HBD-3, heliomicin, alpha-defensin 6 and A9Any one or a combination of at least two of K.
8. The nano-drug of claim 1, wherein the glycopeptide drug comprises any one or a combination of at least two of vancomycin derivatives, norvancomycin derivatives, teicoplanin derivatives, ereomycin, ramoplanin, oritavancin, and daptomycin.
9. The nano-drug of claim 1, wherein the glycopeptide drug is oritavancin.
10. The nano-drug of claim 1, wherein the dosage form of the nano-drug comprises a solution or a dry powder.
11. The nano-drug of claim 1, wherein the route of administration of the nano-drug comprises intravenous injection or topical spray.
12. A method of preparing a nano-drug according to any one of claims 1 to 11, comprising: and adding the amphiphilic cyanine dye and a medicine for treating gram-positive bacteria into a buffer solution to obtain the nano-medicine.
13. The method for preparing nano-drugs according to claim 12, wherein the molar weight ratio of the amphiphilic cyanine dye in the buffer solution to the drug for treating gram-positive bacteria is 3:1-20: 1.
14. The method for preparing nano-drugs according to claim 13, wherein the molar weight ratio of the amphiphilic cyanine dye in the buffer solution to the drug for treating gram-positive bacteria is 10: 1.
15. The method for preparing nano-drugs according to claim 12, wherein the amphiphilic cyanine dye and the drug for treating gram-positive bacteria are added by a syringe pump.
16. The method for preparing a nano-drug according to claim 12, wherein the amphiphilic cyanine dye and the drug for treating gram-positive bacteria are simultaneously added to the buffer solution.
17. The method for preparing nano-drug according to claim 12, wherein the amphiphilic cyanine dye and the drug for treating gram-positive bacteria are added to the buffer solution while stirring.
18. The method for preparing a nano-drug according to claim 12, wherein the buffer solution comprises any one or a combination of at least two of pure water, a phosphate buffer solution, a Tris-HCl buffer solution and a HEPES buffer solution.
19. The method for preparing nano-drug according to claim 12, wherein the amphiphilic cyanine dye and the gram-positive drug are added to a buffer solution, and then the free gram-positive drug is removed by dialysis.
20. Use of a nano-drug according to any one of claims 1 to 11 for the preparation of a material or a medicament for the treatment of gram-positive bacteria.
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