CN111001000B - Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof - Google Patents

Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof Download PDF

Info

Publication number
CN111001000B
CN111001000B CN201911258855.3A CN201911258855A CN111001000B CN 111001000 B CN111001000 B CN 111001000B CN 201911258855 A CN201911258855 A CN 201911258855A CN 111001000 B CN111001000 B CN 111001000B
Authority
CN
China
Prior art keywords
feag
nano
alloy
liposome
drug release
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
Application number
CN201911258855.3A
Other languages
Chinese (zh)
Other versions
CN111001000A (en
Inventor
李俊琳
郝凌云
张小娟
梁栋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jinling Institute of Technology
Original Assignee
Jinling Institute of Technology
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Jinling Institute of Technology filed Critical Jinling Institute of Technology
Priority to CN201911258855.3A priority Critical patent/CN111001000B/en
Publication of CN111001000A publication Critical patent/CN111001000A/en
Application granted granted Critical
Publication of CN111001000B publication Critical patent/CN111001000B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0042—Photocleavage of drugs in vivo, e.g. cleavage of photolabile linkers in vivo by UV radiation for releasing the pharmacologically-active agent from the administered agent; photothrombosis or photoocclusion
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1277—Processes for preparing; Proliposomes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B22—CASTING; POWDER METALLURGY
    • B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
    • B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
    • B22F1/05—Metallic powder characterised by the size or surface area of the particles
    • B22F1/054—Nanosized particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B22—CASTING; POWDER METALLURGY
    • B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
    • B22F9/00—Making metallic powder or suspensions thereof
    • B22F9/16—Making metallic powder or suspensions thereof using chemical processes
    • B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
    • B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Nanotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Physics & Mathematics (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
  • General Physics & Mathematics (AREA)
  • Manufacturing & Machinery (AREA)
  • Inorganic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medical Informatics (AREA)
  • Composite Materials (AREA)
  • Materials Engineering (AREA)
  • Dispersion Chemistry (AREA)
  • Medicinal Preparation (AREA)

Abstract

The invention prepares a berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release by a supercritical carbon dioxide method, the liposome has dual controlled release functions, namely light-controlled drug release and magnetic-controlled drug release, and the encapsulated FeAg nano-alloy can be used as a photo-thermal switch of the light-controlled drug release and a magneto-thermal switch of the magnetic-controlled drug release to control the drug release of the liposome. In practical application, the drug release method can be selected according to requirements. If the drug is required to be released quickly, the controlled release drug can be selected, and the content of Ag in the FeAg nano alloy is increased, so that the controlled release rate of the controlled release drug of the FeAg nano alloy liposome is improved. If the medicine needs to be slowly released, the medicine can be slowly released by magnetic control, so that the medicine can be slowly released according to the need, and the liposome can achieve the functions of controlled release and sustained release.

Description

Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof
Technical Field
The invention discloses a preparation method and performance of berberine hydrochloride FeAg nano-alloy liposome with light control performance and magnetic control performance, belonging to the nano field.
Background
Controlled release of drugs has received increasing attention from researchers over the last several years. The controlled release refers to a preparation which actively releases the drug from the preparation at a certain speed and constant speed to an action organ or a specific target tissue within a predetermined time by changing the structure of the preparation through physical, chemical and other methods, and maintains the drug concentration within an effective concentration for a long time, namely, the controlled release preparation has two characteristics of slow release and controlled release. Liposomes, as a nanovesicle with a bilayer structure and a drug carrier with excellent performance, are applied to more and more clinical applications. Among them, achieving controlled release of drugs in liposomes is one of the hot spots of research. Nowadays, most researches are carried out on photosensitive liposome, namely, photosensitive additives are added into the liposome, so that the photosensitive additives in the liposome absorb light energy under special illumination conditions, the light energy is converted into heat energy, the local temperature of the liposome is increased, the liposome reaches the low critical phase transition temperature, phase transition is carried out, and medicines are released. Noble metals, such as gold and silver, have been used in the study of liposome light-controlled drug release.
Nowadays, another common functional material, magnetic nanoparticles, is also paid attention by controlled release researchers, and is mainly applied to biomedical applications, such as magnetic drug targeting and remote magnetic controlled drug release. Researchers have found that magnetic materials can release some heat when an external magnetic field is applied. According to the point, the magnetic nano material can be added into the liposome, and under the action of an external magnetic field, the magnetic material releases a certain amount of heat, so that the temperature of the liposome is raised to reach the low critical phase transition temperature of the liposome, and the drug is released by phase transition.
In summary, the FeAg nano alloy prepared by the invention has the optical property of noble metal and the magnetic property of iron, is wrapped in a bilayer of liposome, takes berberine hydrochloride as a model drug, inspects the appearance and the property of the FeAg nano alloy liposome, respectively studies the light-controlled drug release and the magnetic-controlled drug release of the FeAg nano alloy liposome through an illumination experiment and a magnetic control experiment, compares the advantages and the disadvantages of the two, and lays a theoretical foundation for the clinical application of the light-controlled drug release and the magnetic-controlled drug release of the liposome.
Disclosure of Invention
The invention aims to develop a liposome with light-controlled-magnetron drug release property and wrapped with magnetic nanoparticles. In order to realize the aim, the invention discloses a preparation method of berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic control drug release.
The technical scheme of the invention is as follows:
a preparation method of berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release mainly adopts a microemulsion method to prepare FeAg nano-alloy, and then prepares the berberine hydrochloride FeAg nano-alloy liposome by a supercritical carbon dioxide method, and comprises the following specific steps:
step 1, preparation of FeAg nano alloy: the FeAg nano alloy is prepared by adopting a reverse microemulsion method, 2-ethylhexyl succinate sodium sulfonate (AOT) is used as a surfactant, n-heptane is used as a continuous phase, and a stable AOT n-heptane solution with the concentration of 0.2-0.4 mol/L is prepared. The precursor concentrations are respectively: 0.1-0.2 mol/L of ferric nitrate nonahydrate aqueous solution, 0.1-0.2 mol/L of silver nitrate aqueous solution and 1-1.5 mol/L of sodium borohydride aqueous solution prepared by ice water. Taking a certain volume of prepared AOT n-heptane solution, dropwise adding ferric nitrate aqueous solution into the solution, and fully stirring under the protection of nitrogen until a stable reversed microemulsion system is formed. Transferring the reversed-phase microemulsion containing the ferric nitrate into a three-neck flask, fixing the reversed-phase microemulsion on a mechanical stirrer, dripping a sodium borohydride aqueous solution into the rapidly-stirred reversed-phase microemulsion of the ferric nitrate under the protection of nitrogen, immediately adding a certain amount of silver nitrate aqueous solution after the color is changed, and continuously stirring for 1-1.5 hours until the reaction is complete. After the reaction is finished, adding excessive methanol into the reaction solution to remove excessive AOT, centrifuging at a high rotating speed of 10000 r/min, washing with absolute ethyl alcohol, repeating the operation for several times after ultrasonic dispersion, and finally putting FeAg alloy-NPs into a vacuum drying oven for drying for later use. In the reaction process, fixing the molar ratio (omega) of water/AOT of the whole reaction system to be 3, and changing the molar ratio (R) of Fe3+ and Ag +, so as to prepare FeAg nano alloys with omega =3, R =1:1, 1:5 and 3:1 respectively.
Step 2, preparation of berberine hydrochloride FeAg nano-alloy liposome:
weighing a certain amount of lecithin and cholesterol according to the mass ratio of 3:1-4:1, completely dissolving the lecithin and cholesterol in a round-bottom flask by using 10-12mL of chloroform-methanol solution with the volume ratio of 2:1-2.5, then adding 10-12mL of FeAg nano alloy chloroform-methanol (with the volume ratio of 2: 1-2.5) solution, controlling proper pressure at 40 ℃, and performing rotary evaporation to volatilize the solvent so that the sample is in a uniform film on the bottle wall of the round-bottom flask; continuing to vacuumize, then putting the round-bottom flask attached with the sample film into a constant-temperature vacuum drying oven, and drying for 6-6.5 h; taking a dry sample obtained by rotary evaporation, adding 10-12mL of berberine solution with concentration of 1-1.5 mg/mL under electromagnetic stirring for dissolving, then injecting the mixed solution into an autoclave, sealing the autoclave and putting the autoclave into a constant temperature water bath, introducing CO2Allowing the pressure to reach 16 Mpa, maintaining the pressure at about 15.5-16.5 Mpa, incubating for 30-35min, slowly releasing scCO2, and taking out the sample in the kettle; 3000r min-1Centrifuging for 3-5min to remove unencapsulated FeAg nano alloy to obtain berberine hydrochloride FeAg nano alloy liposome suspension.
In the step 2, the incubation temperature is 42 ℃, the carbon dioxide pressure is kept at 16.00MPa, and the incubation time is 30 min. The lecithin is 10.0 mg/mL-1Cholesterol concentration 3.3 mg/mL-1. The berberine hydrochloride concentration is 1.0 mg/mL-1 。
Preparation of berberine hydrochloride FeAg nano-alloy liposomes with different FeAg nano-alloy addition amounts: according to the preparation method in the step 2, the fixed lecithin concentration is 10-11 mg.mL-1And the cholesterol concentration is 3.3-3.5 mg-1And the concentration of berberine hydrochloride is 1-1.2 mg.mL-1The FeAg nanoalloy (Fe/Ag molar ratio 1: 1) added to the liposomes had a concentration of 36-108. mu. mol/L (for ease of discussion, below)All expressed as C0), berberine hydrochloride and FeAg nano-alloy liposomes with different FeAg nano-alloy addition amounts are respectively prepared.
Preparing berberine hydrochloride FeAg nano-alloy liposomes with different Fe/Ag molar ratios: according to the preparation method in the step 2, the fixed lecithin concentration is 10-11 mg.mL-1And the cholesterol concentration is 3.3-3.5 mg-1And the concentration of berberine hydrochloride is 1-1.2 mg.mL-1And adding FeAg nano alloys with R =1:1, 1:5 and 3:1 to respectively prepare berberine hydrochloride FeAg nano alloy liposomes with different Fe/Ag molar ratios.
The berberine hydrochloride FeAg nano-alloy liposome prepared by the experiment adopts a Transmission Electron Microscope (TEM) to observe the appearance of a sample.
Advantageous effects
1. The berberine hydrochloride FeAg nano-alloy liposome is mainly prepared by a supercritical carbon dioxide method, and the light-controlled drug release rate and the magnetic-controlled drug release rate of the berberine hydrochloride FeAg nano-alloy liposome are different according to different FeAg nano-alloy addition amounts and different Fe/Ag molar ratios. The drug release degree under different conditions can be controlled by regulating the addition amount of the FeAg nano alloy and the Fe/Ag molar ratio, and the method is suitable for research and production.
2. The invention has both light-controlled release performance and magnetic-controlled release performance, and both the light-controlled release and the magnetic-controlled release are influenced by the proportion of Fe and Ag. The performance parameters of the optically controlled release and the magnetically controlled release are obtained through an illumination experiment and a magnetic control experiment.
3. The invention prepares a berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release by a supercritical carbon dioxide method, the liposome has dual controlled release functions, namely light-controlled drug release and magnetic-controlled drug release, and the encapsulated FeAg nano-alloy can be used as a photo-thermal switch of the light-controlled drug release and a magneto-thermal switch of the magnetic-controlled drug release to control the drug release of the liposome. In practical application, the drug release method can be selected according to needs. If the drug is required to be released quickly, the controlled release drug can be selected, and the content of Ag in the FeAg nano alloy is increased, so that the controlled release rate of the controlled release drug of the FeAg nano alloy liposome is improved. If the medicine needs to be slowly released, the medicine can be slowly released by magnetic control, so that the medicine can be slowly released according to the need, and the liposome can achieve the functions of controlled release and sustained release.
Drawings
FIG. 1 is an electron microscope photograph of berberine hydrochloride FeAg nano-alloy liposome synthesized by the invention and common berberine hydrochloride liposome dyed by a sodium phosphotungstate solution;
reference numbers in the figures: (a) a TEM picture of a common berberine hydrochloride liposome after positive dyeing (b) a TEM picture of a berberine hydrochloride FeAg nano-alloy liposome after negative dyeing.
FIG. 2 is a time-dependent change curve of the drug release rate of the berberine hydrochloride FeAg nano-alloy liposome respectively under the irradiation of visible light and ultraviolet light.
FIG. 3 is a graph showing the change of the release rate of berberine hydrochloride FeAg nano-alloy liposomes with different FeAg nano-alloy additions under UV irradiation at different times.
FIG. 4 is a curve showing the drug release rate of berberine hydrochloride FeAg nano-alloy liposomes synthesized by the present invention with different Fe/Ag molar ratios under UV irradiation at different times.
FIG. 5 is a graph showing the variation of the drug release rate of berberine hydrochloride FeAg nano-alloy liposomes of different FeAg nano-alloy additions synthesized by the invention under the action of a magnetic field for different times.
FIG. 6 is a graph showing the variation of the drug release rate of berberine hydrochloride FeAg nano-alloy liposomes synthesized by the present invention with different Fe/Ag molar ratios at different times under the action of a magnetic field.
FIG. 7 is a light-controlled release curve and a magnetron release curve of the berberine hydrochloride FeAg nano-alloy liposome synthesized by the invention;
reference numbers in the figures: (a) a release curve of the common berberine hydrochloride liposome under the action of ultraviolet light (b) a release curve of the berberine hydrochloride FeAg nano-alloy liposome under the action of a magnetic field.
Detailed Description
For the purpose of enhancing the understanding of the present invention, the present invention will be further described in detail with reference to the following examples and the accompanying drawings, which are only used for explaining the present invention and are not to be construed as limiting the scope of the present invention.
Example 1
A preparation method of berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release comprises the following steps:
step 1, preparation of FeAg nano alloy: the FeAg nano alloy is prepared by adopting a reverse microemulsion method, 2-ethylhexyl succinate sodium sulfonate (AOT) is used as a surfactant, n-heptane is used as a continuous phase, and a stable AOT n-heptane solution with the concentration of 0.2-0.4 mol/L is prepared. The precursor concentrations are respectively: 0.1-0.2 mol/L of ferric nitrate nonahydrate aqueous solution, 0.1-0.2 mol/L of silver nitrate aqueous solution and 1-1.5 mol/L of sodium borohydride ice water solution. Transferring the reversed-phase microemulsion containing the ferric nitrate into a three-neck flask, fixing the reversed-phase microemulsion on a mechanical stirrer, dripping a sodium borohydride aqueous solution into the rapidly-stirred reversed-phase microemulsion of the ferric nitrate under the protection of nitrogen, immediately adding a certain amount of silver nitrate aqueous solution after the color is changed, and continuously stirring for 1-1.5 hours until the reaction is complete. After the reaction is finished, adding excessive methanol into the reaction solution to remove excessive AOT, centrifuging at a high rotating speed of 10000 r/min, washing with absolute ethyl alcohol, repeating the operation for several times after ultrasonic dispersion, and finally putting FeAg alloy-NPs into a vacuum drying oven for drying for later use.
Step 2, weighing a certain amount of lecithin (0.1 g) and cholesterol (0.0333 g) according to the mass ratio of 3:1 into a round-bottom flask, completely dissolving the lecithin and cholesterol with 10 mL of chloroform-methanol solution with the volume ratio of 2:1, then adding 10 mL of FeAg nano alloy (water/surfactant molar ratio omega =3, Fe/Ag molar ratio 1: 1) solution prepared in the step 1 into the round-bottom flask, controlling the proper pressure at 40 ℃, and performing rotary evaporation to volatilize the solvent so that the sample presents a uniform film on the bottle wall of the round-bottom flask. Continuing to vacuumize, and then putting the round-bottom flask attached with the sample film into a constant-temperature vacuum drying oven to dry for 6 hours. Taking the dried sample obtained by rotary evaporation, adding 10 mL of berberine hydrochloride solution with the concentration of 1 mg/mL under electromagnetic stirring to dissolve, then injecting the mixed solution into an autoclave, sealing the autoclave and putting the autoclave into a water bath with constant temperature (42 ℃), introducing CO2 to enable the pressure to reach 16 Mpa, maintaining the pressure at about 16 Mpa, and incubating for 30 min. Then slowly releasing scCO2, taking out the sample in the kettle, centrifuging for 3 min at 3000 r.min-1 to remove the unencapsulated FeAg nano alloy, and obtaining the suspension of berberine hydrochloride FeAg nano alloy liposome.
Taking a drop of berberine hydrochloride FeAg nano-alloy liposome prepared by a supercritical carbon dioxide method, then dropwise adding a sodium phosphotungstate coloring agent solution (pH = 5-7), sucking redundant liquid by using filter paper, drying in a vacuum drying oven for 6 hours, and observing by using a transmission electron microscope, wherein the result is shown in figure 1, in the figure, (a) and (b) correspond to an electron microscope photo of the common berberine hydrochloride liposome and the berberine hydrochloride FeAg nano-alloy liposome dyed by a sodium phosphotungstate solution respectively, and the TEM photo shows that the FeAg nano-alloy liposome presents irregular spherical shapes due to coating FeAg nano-alloy, the average grain diameter is close to about 200 nm, and the size of the FeAg nano-alloy liposome is much larger than that of the common liposome. The encapsulated FeAg nano alloy occupies part of the lipid layer space of the liposome, so that the FeAg nano alloy liposome is deformed to different degrees in appearance and is enlarged in size, and the FeAg nano alloy is encapsulated in the liposome to a certain extent.
Example 2
A preparation method of berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release comprises the following steps:
step 1, preparation of FeAg nano alloy: the FeAg nano alloy is prepared by adopting a reverse microemulsion method, 2-ethylhexyl succinate sodium sulfonate (AOT) is used as a surfactant, n-heptane is used as a continuous phase, and a stable AOT n-heptane solution with the concentration of 0.2-0.4 mol/L is prepared. The precursor concentrations are respectively: 0.1-0.2 mol/L of ferric nitrate nonahydrate aqueous solution, 0.1-0.2 mol/L of silver nitrate aqueous solution and 1-1.5 mol/L of sodium borohydride ice water solution. Transferring the reversed-phase microemulsion containing the ferric nitrate into a three-neck flask, fixing the reversed-phase microemulsion on a mechanical stirrer, dripping a sodium borohydride aqueous solution into the rapidly-stirred reversed-phase microemulsion of the ferric nitrate under the protection of nitrogen, immediately adding a certain amount of silver nitrate aqueous solution after the color is changed, and continuously stirring for 1-1.5 hours until the reaction is complete. After the reaction is finished, adding excessive methanol into the reaction solution to remove excessive AOT, centrifuging at a high rotating speed of 10000 r/min, washing with absolute ethyl alcohol, repeating the operation for several times after ultrasonic dispersion, and finally putting FeAg alloy-NPs into a vacuum drying oven for drying for later use. In the reaction process, fixing the molar ratio (omega) of water/AOT of the whole reaction system to be 3, and changing the molar ratio (R) of Fe3+ and Ag +, so as to prepare FeAg nano alloys with omega =3, R =1:1, 1:5 and 3:1 respectively.
Step 2, preparation of berberine hydrochloride FeAg nano-alloy liposome:
weighing a certain amount of lecithin and cholesterol according to the mass ratio of 3:1-4:1, completely dissolving the lecithin and cholesterol in a round-bottom flask by using 10-12mL of chloroform-methanol solution with the volume ratio of 2:1-2.5, then adding 10-12mL of FeAg nano alloy chloroform-methanol (with the volume ratio of 2: 1-2.5) solution, controlling proper pressure at 40 ℃, and performing rotary evaporation to volatilize the solvent so that the sample is in a uniform film on the bottle wall of the round-bottom flask; continuing to vacuumize, then putting the round-bottom flask attached with the sample film into a constant-temperature vacuum drying oven, and drying for 6-6.5 h; taking a dried sample obtained by rotary evaporation, adding 10-12mL of berberine hydrochloride solution with the concentration of 1-1.5 mg/mL under electromagnetic stirring to dissolve, then injecting the mixed solution into an autoclave, sealing the autoclave and putting the autoclave into a constant-temperature water bath, introducing CO2 to ensure that the pressure reaches 16 MPa and is kept at about 15.5-16.516 MPa, incubating for 30-35min, then slowly releasing scCO2, and taking out the sample in the autoclave; centrifuging at 3000r min-1 for 3-5min to remove unencapsulated FeAg nano alloy to obtain berberine hydrochloride FeAg nano alloy liposome suspension;
preparing berberine hydrochloride FeAg nano-alloy liposomes with different FeAg nano-alloy (Fe/Ag molar ratio of 1: 5) adding amounts (36-108 mu mol/L) according to the method, and then preparing the berberine hydrochloride FeAg nano-alloy liposomes (adding amount of 108 mu mol/L) with different Fe/Ag molar ratios (1: 5,1:3, 1:1, 3: 1). Placing berberine hydrochloride FeAg nano-alloy liposome with determined encapsulation efficiency under 80W ultraviolet light (254 nm) (room temperature), irradiating for 5-30 min, placing 1 mL of FeAg nano-alloy liposome suspension into a treated dialysis bag, dialyzing with 100 mL of distilled water for four times at 4 ℃ for 12 h, wherein the dialyzed external liquid is released drug solution, and mixing the solutions dialyzed for four times and fixing the volume. Using distilled water as reference, measuring absorbance value of free berberine hydrochloride at 345 nm, calculating berberine hydrochloride concentration according to standard curve, and calculating drug release degree at different time according to formula release rate C release/C encapsulation × 100% (wherein C release is berberine hydrochloride amount released by liposome, and C encapsulation is total berberine hydrochloride amount encapsulated by liposome).
As shown in FIG. 2, under the irradiation of visible light, the drug release of the berberine hydrochloride FeAg nano-alloy liposome is almost 0. Under the irradiation of ultraviolet light, the drug release rate of the berberine hydrochloride FeAg nano-alloy liposome reaches 35% in 10 min. The FeAg nano alloy can absorb ultraviolet light energy and convert the absorbed light energy into heat energy, so that the temperature of the liposome is increased to reach the low critical phase transition temperature of the liposome, and the medicine is released.
As can be seen from fig. 3, the reason why the FeAg nano-alloy liposome can release the drug under the ultraviolet irradiation condition is the presence of the FeAg nano-alloy. The larger the addition amount of the FeAg nano alloy is, the more ultraviolet energy can be absorbed, so that the FeAg nano alloy can be converted into more heat, and the liposome can release more medicaments.
As can be seen from FIG. 4, the amount of Ag directly affects the drug release effect of the FeAg nano-alloy liposome, and the smaller the Fe/Ag molar ratio, i.e., the lower the Fe content, the higher the Ag content, and the higher the drug release rate of the FeAg nano-alloy liposome.
Example 3
Berberine hydrochloride FeAg nano-alloy liposomes with different FeAg nano-alloy (Fe/Ag molar ratio of 3: 1) adding amounts (36-108 mu mol/L) are prepared according to the preparation method in the example 2, and the berberine hydrochloride FeAg nano-alloy liposomes (adding amount of 108 mu mol/L) with different Fe/Ag molar ratios (1: 5,1:3, 1:1, 3: 1) are prepared according to the preparation steps. Placing the berberine hydrochloride FeAg nano-alloy liposome with the determined encapsulation efficiency in an external magnetic field (room temperature), controlling the placing time to be 0.25-6 hours, then placing 1 mL of FeAg nano-alloy liposome suspension into a treated dialysis bag, dialyzing for 12 hours at 4 ℃ for four times by using 100 mL of distilled water, wherein the dialyzed external liquid is the released drug solution, and mixing the solutions dialyzed for four times and then fixing the volume. Using distilled water as reference, measuring absorbance value of free berberine hydrochloride at 345 nm, calculating berberine hydrochloride concentration according to standard curve, and calculating drug release degree at different time according to formula release rate C release/C encapsulation × 100% (wherein C release is berberine hydrochloride amount released by liposome, and C encapsulation is total berberine hydrochloride amount encapsulated by liposome).
As shown in FIG. 5, when the addition amount of the FeAg nano alloy is 0, the common berberine hydrochloride liposome can release no drug under the action of the magnetic field for 6 h. The drug release of the berberine hydrochloride FeAg nano-alloy liposome under the action of the magnetic field is slow, the release can be basically finished after 6 hours, but the release rate is high. When the addition amount is 108 mu M, the drug release rate of the berberine hydrochloride FeAg nano-alloy liposome reaches 40 percent in 30 min. The release rate remained substantially stable after 6 h of magnetic field, indicating that the drug release was complete, at which point the drug release rate reached 90%. We also find that the drug release rate of the berberine hydrochloride FeAg nano-alloy liposome under the action of the magnetic field is reduced along with the reduction of the addition amount of the FeAg nano-alloy. The experiment shows that the common berberine hydrochloride can not release the medicine under the action of the magnetic field. Due to the FeAg nano alloy, the berberine hydrochloride FeAg nano alloy liposome can convert magnetic field energy into heat energy under the action of a magnetic field, releases the heat energy, raises the temperature of the liposome, reaches a phase transition temperature to release a medicament, and correspondingly reduces the medicament release rate of the corresponding liposome along with the reduction of the addition amount of the FeAg nano alloy.
As can be seen from FIG. 6, the release rate of the berberine hydrochloride FeAg nano-alloy liposome in the magnetic field decreases with the decrease of the Fe/Ag molar ratio. Especially when the Fe/Ag molar ratio is 0, i.e. the liposome is wrapped by pure Ag particles without the presence of Fe, the liposome hardly releases the drug under the action of the magnetic field. The berberine hydrochloride FeAg nano alloy liposome can release the medicine under the action of the magnetic field because the FeAg nano alloy contains Fe, so that the FeAg nano alloy has super paramagnetic property, and under the action of the magnetic field, the magnetic energy can be converted into heat energy to increase the temperature of the liposome so as to generate phase change to release the medicine. Therefore, the content of Fe directly influences the drug release effect of the berberine hydrochloride FeAg nano-alloy liposome under the action of the magnetic field, and the larger the Fe/Ag molar ratio is, the stronger the drug release capability of the berberine hydrochloride FeAg nano-alloy liposome in the magnetic field is.
Example 4
According to the berberine hydrochloride FeAg nano-alloy liposome prepared by the invention, the bilayer is coated with the FeAg nano-alloy with both magnetic and optical properties, so that the liposome can be subjected to light-operated drug release by ultraviolet illumination due to the photo-thermal conversion effect of Ag; and the magnetic heat conversion effect of superparamagnetic Fe can realize magnetic control drug release on the liposome under the action of a magnetic field. Therefore, FeAg nano alloy liposomes with the same FeAg nano alloy addition amount and the same Fe/Ag molar ratio are selected, and the respective drug release effects are compared and analyzed. The FeAg nano-alloy is prepared according to the preparation method of the embodiment 2, the Fe/Ag molar ratio of the FeAg nano-alloy is 1: 1), the adding amount of berberine hydrochloride FeAg nano-alloy liposome is 108 mu mol/L, the berberine hydrochloride FeAg nano-alloy liposome with the determined encapsulation efficiency is placed in ultraviolet light (0-40 minutes) and an external magnetic field (0-6 hours) (room temperature), then 1 mL of FeAg nano-alloy liposome suspension is placed in a treated dialysis bag, 100 mL of distilled water is used for dialysis for 12 hours at 4 ℃ for four times, the dialyzed external liquid is the released drug solution, and the solutions dialyzed for four times are mixed and then subjected to volume fixing. Using distilled water as reference, measuring absorbance value of free berberine hydrochloride at 345 nm, calculating berberine hydrochloride concentration according to standard curve, and calculating drug release degree at different time according to formula release rate C release/C encapsulation × 100% (wherein C release is berberine hydrochloride amount released by liposome, and C encapsulation is total berberine hydrochloride amount encapsulated by liposome).
As can be seen from FIG. 7-a, the FeAg nano-alloy liposome is released after 5min under UV irradiation, the drug release rate can reach 40%, and the drug release rate is almost kept unchanged after 20 min, which indicates that the drug release process is completed. From the figure 7-b, it can be known that the FeAg nano-alloy liposome has a drug release rate of only 20% when acted by a magnetic field for 30min, the drug release rate is continuously increased within 2 h under the action of the magnetic field, the drug release rate is almost unchanged after 4 h, and the drug release rate can reach more than 80%. Therefore, the ultraviolet light-controlled drug release rate of the FeAg nano alloy liposome is faster, the drug can be released in a short time, and the drug release can be finished within 20 min, but the drug release rate is not high and can only reach 45 percent as can be seen by comparing the analysis of figures 7-a and 7-b. The FeAg nano-alloy liposome has slower magnetic control drug release rate, the whole drug release process needs about 4 hours, but the drug release rate is higher and can reach nearly 80 percent of the drug release rate.

Claims (10)

1. A preparation method of berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release is characterized by comprising the following steps:
step 1, preparing the FeAg nano alloy by a microemulsion method:
and 2, preparing the berberine hydrochloride FeAg nano-alloy liposome by a supercritical carbon dioxide method.
2. The preparation method of the berberine hydrochloride FeAg nano-alloy liposome with the functions of optically controlled drug release and magnetically controlled drug release according to claim 1, wherein the step 1 comprises the following specific steps:
accurately weighing a certain amount of 2-ethylhexyl sulfosuccinate AOT, fully dissolving the 2-ethylhexyl sulfosuccinate AOT in n-heptane, and preparing a stable AOT n-heptane solution with the concentration of 0.2-0.4 mol/L;
accurately preparing 0.1-0.2 mol/L ferric nitrate nonahydrate aqueous solution, 0.1-0.2 mol/L silver nitrate aqueous solution and 1-1.5 mol/L sodium borohydride aqueous solution prepared by ice water respectively in the step (12);
step (13) taking a certain volume of prepared AOT (argon oxygen decarburization) n-heptane solution, dropwise adding a ferric nitrate nonahydrate aqueous solution into the AOT n-heptane solution, and fully stirring the solution under the protection of nitrogen until a stable ferric nitrate reversed-phase micro-emulsion system is formed;
transferring the reversed-phase microemulsion containing the ferric nitrate into a three-neck flask, fixing the reversed-phase microemulsion on a mechanical stirrer, dripping a sodium borohydride aqueous solution into the rapidly-stirred reversed-phase microemulsion of the ferric nitrate under the protection of nitrogen, immediately adding a certain amount of silver nitrate aqueous solution after the color is changed, and continuously stirring for 1-1.5 hours until the reaction is complete;
and (15) after the reaction is finished, adding excessive methanol into the reaction liquid to remove excessive AOT, centrifuging at a high rotating speed of 10000 r/min, washing with absolute ethyl alcohol, repeating the operation for several times after ultrasonic dispersion, and finally drying the FeAg nano alloy in a vacuum drying oven for later use.
3. The preparation method of the berberine hydrochloride FeAg nano-alloy liposome with the functions of optically controlled drug release and magnetically controlled drug release according to claim 1, wherein the step 2 comprises the following specific steps:
weighing a certain amount of lecithin and cholesterol in a round-bottom flask according to the mass ratio of 3:1-4:1, and adding 10-12mL of lecithin and cholesterol in a volume ratio of 2:1-2.5 of chloroform-methanol solution is completely dissolved, and then 10-12mL of chloroform-methanol with the volume ratio of 2:1-2.5 of FeAg nano alloy solution, controlling proper pressure at 40 ℃, and performing rotary evaporation to volatilize the solvent so that the sample presents a layer of uniform film on the bottle wall of the round-bottom flask;
continuously vacuumizing the step (22), and then putting the round-bottom flask attached with the sample film into a constant-temperature vacuum drying oven to dry for 6-6.5 h; taking a dry sample obtained by rotary evaporation, adding 10-12mL of berberine hydrochloride solution with concentration of 1-1.5 mg/mL under electromagnetic stirring to dissolve, then injecting the mixed solution into an autoclave, sealing the autoclave and putting the autoclave into a constant temperature water bath, introducing CO2Allowing the pressure to reach 16 Mpa, maintaining the pressure at 15.5-16.5 Mpa, incubating for 30-35min, and slowly releasing scCO2Taking out the sample in the kettle;
step (23) sample is put at 3000 r.min-1Centrifuging for 3-5min to remove non-encapsulatedFeAg nano alloy to obtain suspension of berberine hydrochloride and FeAg nano alloy liposome.
4. The method for preparing the berberine hydrochloride FeAg nano-alloy liposome with the optically-controlled drug release and the magnetically-controlled drug release functions as claimed in claim 3, wherein in the step (23), the concentration of the FeAg nano-alloy in the suspension of the berberine hydrochloride FeAg nano-alloy liposome is 36-108 μmol/L.
5. The method for preparing the berberine hydrochloride FeAg nano-alloy liposome with the optically-controlled drug release and the magnetically-controlled drug release functions as claimed in claim 3, wherein in the step (22), the incubation temperature is 42 ℃, the carbon dioxide pressure is kept at 16.00MPa, and the incubation time is 30 min.
6. The method for preparing the berberine hydrochloride FeAg nano-alloy liposome with the optically-controlled drug release and the magnetically-controlled drug release functions as claimed in claim 3, wherein in the step (21), the lecithin is 10.0 mg-mL-1Cholesterol concentration 3.3 mg/mL-1。
7. The method for preparing the berberine hydrochloride FeAg nano-alloy liposome with the optically-controlled drug release and the magnetically-controlled drug release functions as claimed in claim 3, wherein in the step (22), the berberine hydrochloride concentration is 1.0 mg-mL-1 。
8. The method for preparing berberine hydrochloride FeAg nano-alloy liposome with light-operated drug release and magnetron drug release as claimed in claim 2, wherein in the step 1, the molar ratio omega of water/AOT for fixing the whole reaction system is 3, and Fe is changed3+And Ag+With a molar ratio of R, the FeAg nanoalloys were prepared with R =1:1, 1:5 and 3: 1.
9. The method for preparing berberine hydrochloride FeAg nano-alloy liposome with light-controlled release and magnetron release of drugs according to claim 1, which comprisesCharacterized in that, in the step 1, the liquid reagent to be used is subjected to N bubbling before use2The oxygen is removed first.
10. The berberine hydrochloride FeAg nano-alloy liposome with the functions of optically controlled drug release and magnetically controlled drug release, which is prepared by the preparation method of any one of the claims 1 to 9.
CN201911258855.3A 2019-12-10 2019-12-10 Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof Active CN111001000B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201911258855.3A CN111001000B (en) 2019-12-10 2019-12-10 Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201911258855.3A CN111001000B (en) 2019-12-10 2019-12-10 Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof

Publications (2)

Publication Number Publication Date
CN111001000A CN111001000A (en) 2020-04-14
CN111001000B true CN111001000B (en) 2022-02-01

Family

ID=70114363

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911258855.3A Active CN111001000B (en) 2019-12-10 2019-12-10 Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof

Country Status (1)

Country Link
CN (1) CN111001000B (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101683322A (en) * 2008-09-26 2010-03-31 华东理工大学 Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006051732A1 (en) * 2004-11-10 2006-05-18 Konica Minolta Medical & Graphic, Inc. Pharmaceutical preparation containing coated magnetic particles and method for production thereof, and diagnosis therapy system

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101683322A (en) * 2008-09-26 2010-03-31 华东理工大学 Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"盐酸小檗碱磁向光敏脂质体的制备及性质研究";王冬青;《中国优秀硕士学位论文全文数据库工程科技I辑》;20150715(第07期);第1-89页 *
T. J. JACKSON等."Superparamagnetism in Heterogeneous AgFe Thin Films – A Low Energy μSR Study".《Hyperfine Interactions》.2001,第403-408页. *

Also Published As

Publication number Publication date
CN111001000A (en) 2020-04-14

Similar Documents

Publication Publication Date Title
CN111265533B (en) Preparation method of core-shell nano-particles based on lipid membrane and metal organic framework
Zhao et al. Colorimetric detection of blood glucose based on GOx@ ZIF-8@ Fe-polydopamine cascade reaction
CN103100725B (en) Preparation method of silver/carbon quantum dot composite nanometer materials
CN114656645B (en) Preparation method of cyclodextrin metal organic framework stably existing in water phase
CN105776350B (en) A kind of preparation method of hollow bar-shaped ferroso-ferric oxide and its compound
CN111185235B (en) Preparation of gold nanoparticle/metal organic framework compound and application of gold nanoparticle/metal organic framework compound in p-nitrophenol reduction
Qiu et al. Triple-stimuli (protease/redox/pH) sensitive porous silica nanocarriers for drug delivery
CN107998183B (en) Osmanthus fragrans phenylethanoid glycoside liposome coated with chitosan and preparation method thereof
CN107648606A (en) Collect chemotherapy and the preparation method of thermotherapy integrated nuclear shell nanometer material and products thereof and application
CN110026568B (en) Method for synthesizing medicinal dendritic nano gold particles by mediation of polyhydroxy natural products
CN109463751B (en) Preparation method of coenzyme Q10 liposome health food
CN109295716A (en) A kind of preparation method of magnetic, temperature collaboration stimuli responsive hydrogel
Wang et al. Mesostructured TiO2 Gated Periodic Mesoporous Organosilica‐Based Nanotablets for Multistimuli‐responsive Drug Release
Sun et al. A porous material excited by near-infrared light for photo/chemodynamic and photothermal dual-mode combination therapy
Gao et al. Synthesis of ‘dual-key-and-lock’drug carriers for imaging and improved drug release
CN111001000B (en) Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof
CN107970224B (en) Preparation method and application of lipid-modified magnetic graphene oxide composite material
Tian et al. Design of a near‐infrared‐triggered photo/thermal dual‐responsive composite carrier with excellent biocompatibility for controllable drug release
Peng et al. Application and Development Prospect of Nanoscale Iron Based Metal-Organic Frameworks in Biomedicine
CN110961055B (en) Ionic liquid polymeric microspheres and preparation method and application thereof
CN111847524B (en) Fe (Fe) 3 O 4 /Fe 2 O 3 Magnetic heterogeneous nanotube and preparation method thereof
CN116327737A (en) Preparation method of resveratrol microcapsule based on metal-organic framework
CN110205115B (en) Switch-type CQDs @ Ag core-shell nano fluorescent probe and preparation method and application thereof
Li et al. Using magnetic and photic stimuli-responsive liposomes to serve up chemotherapy drugs to cancer cells
CN108014092A (en) A kind of preparation method and application of magnetic oxygenated graphene-nucleoprotamine/sodium carboxymethylcellulose compound

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