CN112121162A - Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof - Google Patents
Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof Download PDFInfo
- Publication number
- CN112121162A CN112121162A CN202010930177.7A CN202010930177A CN112121162A CN 112121162 A CN112121162 A CN 112121162A CN 202010930177 A CN202010930177 A CN 202010930177A CN 112121162 A CN112121162 A CN 112121162A
- Authority
- CN
- China
- Prior art keywords
- microcapsule
- carboxymethyl chitosan
- oil
- shell
- sensitizer
- 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.)
- Pending
Links
Images
Classifications
-
- 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
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
-
- 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
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- 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
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
-
- 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
- A61K47/51—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 non-active ingredient being a modifying agent
- A61K47/54—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 non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/545—Heterocyclic compounds
-
- 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
- A61K47/6921—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 the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6925—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 the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a microcapsule, nanocapsule, microbubble or nanobubble
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/08—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/36—Materials or treatment for tissue regeneration for embolization or occlusion, e.g. vaso-occlusive compositions or devices
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Surgery (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Nanotechnology (AREA)
- Inorganic Chemistry (AREA)
- Materials Engineering (AREA)
- Gynecology & Obstetrics (AREA)
- Pregnancy & Childbirth (AREA)
- Endocrinology (AREA)
- Reproductive Health (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a microcapsule with chemotherapeutic drugs and thermotherapy sensitizers and a preparation method thereof. The diameter of the microcapsule is 0.5 to 250 μm, and the thickness of the shell of the microcapsule is 0.05 to 30 μm. The microcapsule with the chemotherapeutics and the thermotherapy sensitizer comprises a shell and a core material, wherein the shell is made of carboxymethyl chitosan, the core material is wrapped in the shell, the core material comprises the photothermal sensitizer and the chemotherapeutics, and the outer surface of the shell is coupled with targeting molecules. The microcapsule of the invention is prepared according to the following steps: emulsifying, interfacial polymerization, folic acid coupling and washing to obtain the material for wrapping photothermal heating and tumor embolization. The microcapsule prepared by the invention has the advantages of simple process, good repeatability, high production efficiency and easy realization of large-scale production. The microcapsule prepared by the invention has controllable granularity, size and thickness. The shell material contains a large amount of amino groups, and can endow functions such as drug controlled release, antibody coupling and the like. The microcapsule is a carboxymethyl chitosan microcapsule carrier with the advantages of photo-thermal sensitization, chemotherapy drug sustained and controlled release, targeting and good biocompatibility, and has good application prospect when being used as an embolic agent in breast cancer treatment.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a novel sustained-release embolism carboxymethyl chitosan microcapsule carrying chemotherapeutic medicines and thermotherapy sensitizers together, and a preparation method and application thereof.
Background
Breast cancer is one of main malignant tumors threatening the health of women in China, and currently, clinical treatment methods for primary breast cancer mainly adopt surgical excision, embolization, radiotherapy, chemotherapy and the like, but the methods have certain limitations, such as high surgical risk, large side effect, lack of specificity to the tumor, drug resistance and the like in the treatment process, and cannot achieve the purpose of treatment. The traditional treatment method has extremely limited effect on the tumor after the metastasis, which is also a big reason that the fatality rate of the malignant tumor is difficult to effectively control. In view of the complexity of breast cancer tumor biology and the limitations of various therapeutic approaches, comprehensive treatment using multidisciplinary collaboration, with coexistence of multiple therapeutic approaches, has become a consensus in the field of primary breast cancer therapy.
Photothermal therapy is a method of using near infrared light having a strong penetrating power to human tissue as an energy source to concentrate heat at an affected part, thereby achieving the purpose of destroying tumor tissue and treating tumor (Clinical depletion and potential of photothermal and dynamic therapeutics for cancer, Nature Reviews Clinical Oncology, 2020), and has gradually attracted people's attention due to its advantages of high tumor specificity, small wound, few complications, and the like. However, with the continuous and deep clinical research, the problem of photothermal therapy is highlighted. Firstly, the transmission capacity of near infrared light is limited, so that the near infrared light cannot be incident to the deep layer of the tumor, the tumor cannot be killed completely, and the tumor is easy to recur; secondly, the heating range is difficult to be limited to the tumor site, and the damage of normal tissues or organs around the tumor is easily caused. In addition, the photothermal material can be used as a Multifunctional diagnostic agent effective against tumors by contrast by means of surface modification or the like, or by synergistic therapy with chemotherapy, radiotherapy, immunotherapy, or the like (Chemical Society Reviews, 2020). The photothermal therapy based on the micro-nano material is used as a new means for treating the tumor, so that the nano photothermal therapeutic agent actively or passively enriched on the affected part by various targeting technologies generates heat under the irradiation of near infrared light, thereby achieving the purposes of destroying tumor tissues and treating the tumor. Photothermal sensitizer molecules (ACS Applied Materials & Interfaces, 2017) can efficiently absorb near infrared light and convert the near infrared light into heat energy and singlet oxygen, and are used in multiple fields of disease imaging diagnosis, Photothermal Therapy and the like. However, the current drug-carrying systems based on indocyanine green and the like still have the problems of local accumulation, slow metabolism, excessive tissue toxicity and the like, and the research of safe drug systems and highly efficient targeting technologies is still a serious challenge.
The invention discloses a preparation method and application of a photosensitive magnetic nanoparticle system for inhibiting the growth of breast cancer cells, and belongs to the Chinese patent with the application number of 201710035771.8. The test comprises the following main steps: 1. magnetic nano particle Fe wrapped by oleic acid3O4-OA is subjected to amination treatment to activate the carboxyl groups on the photosensitizer; 2. grafting of photosensitizers to Fe by condensation of amino and carboxyl groups3O4-OA surface to obtain the photosensitive magnetic nanoparticle system. The magnetic field has the effect of photodynamic therapy of breast cancer under the illumination condition, realizes the combination of breast cancer targeting and folate cell targeting of a magnetic field, can efficiently and targetedly inhibit the growth of breast cancer cells, and has the characteristics of good slow release, stability, dispersibility and uniformity and low toxic and side effects. However, the photothermal therapeutic agent is not good enough in the aspect of treating malignant tumors and the like, and cannot be used for tumor embolization treatment and/or photothermal treatment and/or chemotherapy at the same time.
Carboxymethyl chitosan is a water-soluble chitosan derivative, has many characteristics, such as strong antibacterial property and fresh-keeping effect, and is an amphoteric polyelectrolyte. The chitosan derivative has various applications in the aspects of cosmetics, fresh keeping, medicines and the like, and is one of the chitosan derivatives which are researched more recently. The carboxymethyl chitosan is an important shell material for preparing the microcapsule, and has the advantages of good biocompatibility, biodegradability and the like.
At present, aiming at the problems of local accumulation, slow metabolism, overlarge tissue toxicity and the like of a medicine carrying system such as indocyanine green and the like, no research is found for preparing microcapsules carrying chemotherapeutic drugs, indocyanine green and other hyperthermia sensitizers from carboxymethyl chitosan and using the microcapsules in tumor photothermal therapy and chemotherapy.
Disclosure of Invention
The first purpose of the invention is to provide an embolism carboxymethyl chitosan microcapsule compound with good biocompatibility and low toxicity to human bodies, which is used for the functions of light sensitization and the sustained and controlled release of chemotherapeutic drugs.
The second purpose of the invention is to provide a preparation method of the embolism carboxymethyl chitosan microcapsule compound with the functions of light sensitization and chemotherapy drug sustained and controlled release. The size of the microcapsules and the thickness of the shell can be adjusted. The invention has simple process, good repeatability and high production efficiency, and is easy to realize large-scale production.
The third purpose of the invention is to provide the chemotherapy drug and the photothermal sensitizer which are encapsulated in the microcapsule, so that the combination of chemotherapy and photothermal therapy can be realized.
In view of the first object, the present invention adopts the following technical solutions:
the microcapsule comprises a shell, wherein the shell is made of carboxymethyl chitosan, a core material is wrapped in the shell and comprises a chemotherapeutic drug and a photothermal sensitizer, and the outer surface of the shell is coupled with a targeting molecule. The microcapsule is dispersed in water, and generates hyperpyrexia of 10-40 ℃ within 5min, so that tumor cells are thermally killed.
Preferably, the diameter of the microcapsule is 0.5 to 250 μm, and the thickness of the shell of the microcapsule is 0.05 to 30 μm.
Preferably, the chemotherapeutic agent is selected from the group consisting of alkylating agents, antimetabolites, antitumor antibiotics, plant-based anticancer agents, hormones and immunological agents in combination with one or more thereof.
Preferably, the thermotherapy sensitizer is one or more of indocyanine green, porphyrin, phenyl porphyrin and hematoporphyrin monomethyl ether.
Preferably, the targeting molecule is folic acid.
Preferably, the carboxymethyl chitosan has a molecular weight of 2 to 20 ten thousand.
In order to achieve the second object, the invention adopts the following technical scheme:
the preparation method of the microcapsule comprises the following steps:
(1) dissolving carboxymethyl chitosan, chemotherapeutic medicine and photo-thermal sensitizer in water solution to obtain water phase;
(2) then the oil and the emulsifier are stirred and mixed evenly to form an oil phase;
(3) adding the water phase into the oil phase, and stirring to form an emulsion;
(4) adding a cross-linking agent into the obtained emulsion for curing to obtain carboxymethyl chitosan microcapsules;
(5) adding folic acid, NHS and a DMSO solution containing EDC into the obtained carboxymethyl chitosan microcapsule; magnetically stirring overnight at room temperature in the dark; and then centrifuging the liquid sample, and removing redundant small molecular substances such as folic acid, EDC and the like to obtain the folic acid modified carboxymethyl chitosan microcapsule.
Preferably, the concentrations of the chemotherapeutic drug and the photothermal sensitizer in the aqueous phase in the step (1) are 0.01-10 mg/L and 0.05-30 mg/L respectively, and the concentration of the carboxymethyl chitosan in the aqueous phase is 0.01-5 g/mL; and/or stirring at 500-1500 r/min in the step (1) to form a water phase; and/or, the oil in the step (2) is selected from one or more of peanut oil, olive oil, rapeseed oil, corn oil, sesame oil, cottonseed oil, rice oil, sunflower seed oil, linseed oil, safflower seed oil, soybean oil, paraffin, n-octane or isooctane; and/or the emulsifier in the step (2) is selected from one or more of Span80, Span85, Span60, sodium-calcium stearoyl lactylate, sodium stearoyl lactylate, triglyceryl, propylene glycol fatty acid ester, sucrose ester, soybean lecithin, lauric acid monoglyceride, ethylene glycol fatty acid ester or polyoxyethylene monolaurate; and/or stirring at 500-1500 r/min in the step (2) to form an oil phase; and/or in the step (3), the emulsification process is stirring at 500-1500 r/min, the emulsification temperature is 20-50 ℃, and the emulsification time is 20-90 min; and/or, in the step (4), the curing time is 0.5-3 h, and the curing temperature is 30-70 ℃; and/or, the cross-linking agent in the step (4) is selected from one or more of dicumyl peroxide, benzoyl peroxide, di-tert-butyl peroxide, dicumyl peroxide, 2-ethyl-4-methylimidazole, 2-phenylimidazole, hexahydrophthalic anhydride, triethylenetetramine, dimethylaminopropylamine, diethylaminopropylamine, melamine, polyethylene glycol, divinylbenzene, formaldehyde and glutaraldehyde; and/or, in the step (4), the curing is carried out under the stirring of 500-1500 r/min; and/or the dosage ratio of the carboxymethyl chitosan microcapsule, the folic acid, the NHS and the EDC in the step (5) is (0.01-0.2): (0.01-0.2) by molar mass ratio of 1.
In order to achieve the third object, the present invention adopts the following technical solutions:
the microcapsule of the invention is used for preparing antitumor diagnosis and treatment agents.
Preferably, the antitumor medical agent is used for tumor embolism treatment and/or photothermal treatment and/or chemotherapy and/or disease imaging diagnosis.
The carboxymethyl chitosan microcapsule carrying the chemotherapeutic drug and the thermotherapy sensitizer limits the photosensitizer in a certain space, and reduces the toxic and side effects of the thermotherapy sensitizer on normal tissues of a body. After the tumor cell is gathered in the tumor, the temperature can be rapidly raised under the illumination, the photothermal conversion efficiency in the tumor is improved, meanwhile, the chemotherapeutic drug is slowly controlled, and the photothermal and the chemotherapeutic can be synergistically and continuously acted on the breast cancer tumor cells. The surface of the carboxymethyl chitosan microcapsule is coupled with target molecule folic acid, and the carboxymethyl chitosan microcapsule can target breast cancer tumor tissues and can continuously kill residual tumor cells in the tumors. The carboxymethyl chitosan microcapsule with the size of micron preferably selected by the invention can cause the tumor vascular flow retardation after being injected into the artery, thereby achieving the effect of blocking the tumor tissue blood vessel and greatly improving the treatment effect. The carboxymethyl chitosan is easier to release chemotherapeutic drugs after being degraded in a weak acid environment of tumor, can kill tumor cells in a high-efficiency targeted manner, and has good clinical application prospect.
In addition, the carboxymethyl chitosan microcapsule provided by the invention has rich functions, can be synchronously used for tumor embolization treatment, photothermal treatment or chemotherapy after being injected into an artery, and can be used for imaging detection through the light effect of molecules such as indocyanine green and the like. Therefore, the dosage of the medicine for the patient can be reduced, and the pain of the patient can be relieved.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 is a schematic structural diagram of a carboxymethyl chitosan microcapsule provided by the present invention;
FIG. 2 is a flow chart of the preparation of carboxymethyl chitosan microcapsule provided by the present invention;
FIG. 3 is an optical microscope photograph of microcapsules prepared in accordance with an embodiment of the present invention;
FIG. 4 is a photo-thermal temperature-rise curve of the microcapsules in examples 3 to 5 of the present invention;
FIG. 5 is a drug release profile of microencapsulated doxorubicin in examples 3-5 of the present invention;
fig. 6 shows the targeting effect of the carboxymethyl chitosan microcapsule and the folic acid modified carboxymethyl chitosan microcapsule on breast cancer cells in example 2 of the present invention under an optical microscope.
Detailed Description
The raw materials of the invention are all commercial products.
Example 1
Dissolving carboxymethyl chitosan in 5mL of aqueous solution to make the concentration of the carboxymethyl chitosan be 0.1g/mL, and stirring the solution at 500r/min to form a water phase; stirring 20mL of soybean oil and 0.5mL of Span80 emulsifier at 500r/min to form an oil phase; the water phase is added into the oil phase dropwise, and the mixture is stirred at room temperature of 25 ℃ and 500r/min for emulsification for 60 min.
The reaction solution was placed in a hot water bath at 70 ℃ and 0.5mL of 50% glutaraldehyde was added, followed by stirring and curing at 500r/min for 1 hour. After the reaction is finished, the microcapsule is obtained by washing with water and isopropanol and drying, and the size of the obtained microcapsule is about 5 mu m (see figure 3).
Example 2
Dissolving carboxymethyl chitosan and chemotherapeutic drug adriamycin in 10mL of aqueous solution to make the concentrations of the carboxymethyl chitosan and the chemotherapeutic drug adriamycin respectively 0.02g/mL and 100 mu g/mL, and stirring at 1400r/min to form a water phase; stirring 100mL of olive oil and 1.5mL of Span60 emulsifier at 1400r/min to form an oil phase; the aqueous phase was added dropwise to the oil phase and emulsified at 35 ℃ for 40min with stirring at 1400 r/min.
The reaction solution was placed in a hot water bath at 60 ℃ and 0.5mL of 50% formaldehyde was added thereto, followed by stirring and curing at 1400r/min for 2 hours. After the reaction is finished, washing with water and isopropanol respectively, and drying to obtain the microcapsule, wherein the size of the obtained microcapsule is about 500 nm.
Preparing a mixed solution containing 0.05mol/L NHS and 0.025mol/L EDC by taking DMSO as a solvent, taking 50mg of microcapsules, respectively adding 1.0mg of folic acid into 0.1mL of the mixed solution, magnetically stirring overnight at room temperature in the dark, then centrifuging a liquid sample, and removing redundant folic acid, EDC and other small molecular substances to obtain the folic acid modified carboxymethyl chitosan microcapsules.
Example 3
Dissolving carboxymethyl chitosan, chemotherapeutic drug adriamycin and photo-thermal sensitizer indocyanine green in 10mL of aqueous solution to make the concentrations of the carboxymethyl chitosan, the chemotherapeutic drug adriamycin and the photo-thermal sensitizer indocyanine green respectively be 0.1g/mL, 10 mu g/mL and 0.05mg/mL, and stirring at 800r/min to form a water phase; stirring 80mL of linseed oil and 0.8mL of triglycerin emulsifier at 800r/min to form an oil phase; the water phase is added into the oil phase dropwise, and emulsification is carried out at 35 ℃ and 800r/min under stirring for 45 min.
The reaction solution was placed in a hot water bath at 70 ℃ and 0.8mL of glutaraldehyde was added, and the mixture was stirred at 800r/min for 0.5h to cure. After the reaction is finished, washing with water and isopropanol respectively, and drying to obtain the microcapsule, wherein the size of the obtained microcapsule is about 20 mu m.
Preparing a mixed solution containing 0.01mol/L NHS and 0.005mol/L EDC by taking DMSO as a solvent, taking 20mg of microcapsules, respectively adding 0.3mg of folic acid into 0.2mL of the mixed solution, magnetically stirring overnight at room temperature in the dark, then centrifuging a liquid sample, removing redundant folic acid, EDC and other small molecular substances, and obtaining the folic acid modified carboxymethyl chitosan microcapsules.
Testing the photo-thermal performance: dispersing 25mg of the prepared microcapsule in 1mL of pure water, taking 1mL of pure water as a control group, and using a 808nm laser (1W/cm) for the experimental group and the control group2) Irradiating for 5min, respectively, measuring temperature change with digital thermometer (see figure 4 a), raising temperature 16 deg.C higher than pure water
Example 4
Dissolving carboxymethyl chitosan, chemotherapeutic drug adriamycin and photo-thermal sensitizer indocyanine green in 10mL of aqueous solution to make the concentrations of the carboxymethyl chitosan, the chemotherapeutic drug adriamycin and the photo-thermal sensitizer indocyanine green be 1g/mL, 100 mu g/mL and 1.0mg/mL respectively, and stirring at 900r/min to form a water phase; stirring 100mL of peanut oil and 5mL of triglycerin emulsifier at 900r/min to form an oil phase; adding the water phase into the oil phase dropwise, and emulsifying at 30 deg.C and 900r/min under stirring for 45 min.
The reaction mixture was placed in a hot water bath at 70 ℃ and 1.5mL of dicumyl peroxide was added thereto, followed by stirring and curing at 900r/min for 2.5 hours. After the reaction is finished, washing with water and isopropanol respectively, and drying to obtain the microcapsule, wherein the size of the obtained microcapsule is about 100 mu m.
Preparing a mixed solution containing 0.5mol/L NHS and 0.15mol/L EDC by taking DMSO as a solvent, taking 20mg of microcapsules, respectively adding 0.6mg of folic acid into 0.5mL of the mixed solution, magnetically stirring overnight at room temperature in the dark, then centrifuging a liquid sample, removing redundant folic acid, EDC and other small molecular substances, and obtaining the folic acid modified carboxymethyl chitosan microcapsules.
Testing the photo-thermal performance: dispersing 25mg of the prepared microcapsule in 1mL of pure water, taking 1mL of pure water as a control group, and using a 808nm laser (1W/cm) for the experimental group and the control group2) Irradiating for 5min, respectively, measuring temperature change with digital thermometer (see FIG. 4 b), raising temperature 13.5 deg.C higher than pure water
Example 5
Dissolving carboxymethyl chitosan, chemotherapeutic drug adriamycin and photo-thermal sensitizer indocyanine green in 10mL of aqueous solution to make the concentrations of the carboxymethyl chitosan, the chemotherapeutic drug adriamycin and the photo-thermal sensitizer indocyanine green respectively 5g/mL, 200 mu g/mL and 3.0mg/mL, and stirring at 500r/min to form a water phase; stirring 150mL of sunflower seed oil and 5mL of sodium stearoyl lactate emulsifier at 500r/min to form an oil phase; the water phase is added dropwise into the oil phase, and emulsified at 40 deg.C under stirring at 500r/min for 45 min.
The reaction solution was placed in a hot water bath at 60 ℃ and 3mL of benzoyl peroxide was added thereto, followed by stirring and curing at 500r/min for 3 hours. After the reaction is finished, washing with water and isopropanol respectively, and drying to obtain the microcapsule, wherein the size of the obtained microcapsule is about 200 mu m.
Preparing a mixed solution containing 0.9mol/L NHS and 0.45mol/L EDC by taking DMSO as a solvent, taking 20mg of microcapsules and 1.0mg of folic acid, respectively adding 2mL of the mixed solution, magnetically stirring overnight at room temperature in the dark, then centrifuging a liquid sample, and removing redundant folic acid, EDC and other small molecular substances to obtain the folic acid modified carboxymethyl chitosan microcapsules.
Testing the photo-thermal performance: dispersing 25mg of the prepared microcapsule in 1mL of pure water, taking 1mL of pure water as a control group, and using a 808nm laser (1W/cm) for the experimental group and the control group2) The irradiation was carried out for 5min, and the temperature change was measured with a digital thermometer, see FIG. 4c, which increased the temperature 11.6 ℃ higher than that of pure water.
As can be seen from the experimental results of examples 3-5, the microcapsules provided by the invention have good photo-thermal properties.
Example 6
Simulating the drug loading and encapsulation rate of the microcapsules on the adriamycin in vitro by adopting a vibration dialysis method: the drug-loaded microspheres (i.e., microcapsules) containing 25mg of doxorubicin prepared in examples 3-5 were placed in 100mL of PBS buffer solution for in vitro release experiments. Samples were taken at defined time points and the supernatant was uv-vis-spectrographed and the doxorubicin release profile was plotted. The cumulative release at 24 hours for the loaded microspheres in examples 3-5 (corresponding to a, b, and c in FIG. 5, respectively) was 35.3%, 31.4%, and 28%, respectively. The cumulative drug release amount of the microspheres decreases with time and the release duration is long. The microcapsule of the invention has good slow release effect.
Example 7
In vitro targeting experiment of breast cancer: in vitro culture of Breast cancer cell line 4T1 (cell culture conditions: 10% fetal bovine serum, 90% RPMI 1640, 5% CO)237 ℃ C.). And (3) test groups: a carboxymethyl chitosan microcapsule group which is not modified by folic acid is set (the preparation method of the microcapsule is consistent with that of example 2, except that folic acid is not added for modification), and the carboxymethyl chitosan microcapsule group is modified by folic acid (the preparation method of the microcapsule is consistent with that of example 2). The log phase 4T1 breast cancer cells were collected and seeded in 24-well plates. When the cells are completeAfter the adherence, equal amounts of the carboxymethyl chitosan microcapsule prepared by the method of example 2 and the folic acid modified carboxymethyl chitosan microcapsule are respectively added, the incubation is carried out for 2 hours at 37 ℃, then the washing is carried out for 3 times by using PBS liquid, and the coupling condition of the microcapsule and the breast cancer cells is observed by using a laser confocal microscope. Under a laser confocal microscope (figure 6), the folic acid modified carboxymethyl chitosan microcapsule is tightly combined with the breast cancer cells, and the fact that the microcapsule is tightly combined with the breast cancer cells in a targeted manner is proved to have good targeting capability; the experiment result of the carboxymethyl chitosan microcapsule is only less combined with breast cancer cells, and the breast cancer targeting agent on the surface of the microcapsule disclosed by the invention is proved to have the specific targeting combination capability of breast cancer.
Examples 8 to 10
The difference is that the indocyanine green added is replaced by one or more of porphyrin, phenyl porphyrin and hematoporphyrin monomethyl ether as in example 2.
Examples 11 to 12
The difference is that the indocyanine green added is replaced by one or more of porphyrin, phenyl porphyrin and hematoporphyrin monomethyl ether as in example 3.
Examples 11 to 23
The method is the same as example 4, except that the added peanut oil is replaced by one or more of olive oil, rapeseed oil, corn oil, sesame oil, cottonseed oil, rice oil, sunflower oil, linseed oil, safflower oil, soybean oil, paraffin, n-octane or isooctane.
Examples 24 to 34
The method is the same as example 5, except that the sodium stearoyl lactylate is replaced by one or more of Span80, Span85, Span60, sodium stearoyl lactylate-calcium, triglyceride, propylene glycol fatty acid ester, sucrose ester, soybean lecithin, monolaurate, ethylene glycol fatty acid ester or polyoxyethylene monolaurate.
Examples 35 to 40
The difference from example 3 is that doxorubicin is replaced with a combination of one or more of an alkylating agent, an antimetabolite, an antitumor antibiotic, a plant anticancer agent, a hormone, and an immunological agent.
In conclusion, the experiments of examples 3-5 prove that the microcapsules provided by the invention have good photo-thermal properties. The experiment of example 6 shows that the microcapsule of the present application has the effect of slowly releasing chemotherapeutic drugs. The experiment in example 7 proves that the breast cancer targeting agent on the surface of the microcapsule provided by the application has the specific targeting binding capacity for breast cancer. Therefore, the embolism carboxymethyl chitosan microcapsule compound with the light sensitization function and the chemotherapy drug sustained-release function has good application potential in the combined application of chemotherapy and photothermal therapy of malignant tumors.
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention, and it will be obvious to those skilled in the art that other variations or modifications may be made on the basis of the above description, and all embodiments may not be exhaustive, and all obvious variations or modifications may be included within the scope of the present invention.
Claims (10)
1. A sustained-release carboxymethyl chitosan microcapsule carrying chemotherapeutic drugs and thermotherapy sensitizers is characterized in that: the microcapsule comprises a shell and a core material, wherein the shell wraps the core material, the shell is made of carboxymethyl chitosan, the core material is a chemotherapeutic drug and a photothermal sensitizer, and the outer surface of the shell is coupled with a targeting molecule.
2. The slow-release carboxymethyl chitosan microcapsule for co-loading chemotherapeutic drugs and hyperthermia sensitizers according to claim 1, wherein: the diameter of the microcapsule is 0.5 to 250 μm, and the thickness of the shell of the microcapsule is 0.05 to 30 μm.
3. The slow-release carboxymethyl chitosan microcapsule for co-loading chemotherapeutic drugs and hyperthermia sensitizers according to claim 1, wherein: the chemotherapeutic drug is selected from one or more of alkylating agent, antimetabolite, antitumor antibiotic, plant anticancer drug, hormone and immunological preparation.
4. The slow-release carboxymethyl chitosan microcapsule for co-loading chemotherapeutic drugs and hyperthermia sensitizers according to claim 1, wherein: the heat treatment sensitizer is one or more of indocyanine green, porphyrin, phenyl porphyrin and hematoporphyrin monomethyl ether.
5. A thermal therapy sensitizer sustained-release carboxymethyl chitosan microcapsule according to claim 2, characterized in that: the targeting molecule is folic acid.
6. A thermal therapy sensitizer sustained-release carboxymethyl chitosan microcapsule according to claim 2, characterized in that: the molecular weight of the carboxymethyl chitosan is 2-20 ten thousand.
7. A process for the preparation of microcapsules according to claim 1, comprising the steps of:
(1) dissolving carboxymethyl chitosan, chemotherapeutic medicine and photo-thermal sensitizer in water solution to obtain water phase;
(2) then the oil and the emulsifier are stirred and mixed evenly to form an oil phase;
(3) adding the water phase into the oil phase, and stirring to form an emulsion;
(4) adding a cross-linking agent into the obtained emulsion for curing to obtain carboxymethyl chitosan microcapsules;
(5) adding folic acid, NHS and a DMSO solution containing EDC into the obtained carboxymethyl chitosan microcapsule; magnetically stirring overnight at room temperature in the dark; and then centrifuging the liquid sample to obtain the folic acid modified carboxymethyl chitosan microcapsule.
8. The method of claim 4, wherein: the concentrations of the chemotherapeutic drug and the photothermal sensitizer in the aqueous phase in the step (1) are respectively 0.01-10 mg/L and 0.05-30 mg/L, and the concentration of the carboxymethyl chitosan in the aqueous phase is 0.01-5 g/mL; and/or stirring at 500-1500 r/min in the step (1) to form a water phase; and/or, the oil in the step (2) is selected from one or more of peanut oil, olive oil, rapeseed oil, corn oil, sesame oil, cottonseed oil, rice oil, sunflower seed oil, linseed oil, safflower seed oil, soybean oil, paraffin, n-octane or isooctane; and/or the emulsifier in the step (2) is selected from one or more of Span80, Span85, Span60, sodium-calcium stearoyl lactylate, sodium stearoyl lactylate, triglyceryl, propylene glycol fatty acid ester, sucrose ester, soybean lecithin, lauric acid monoglyceride, ethylene glycol fatty acid ester or polyoxyethylene monolaurate; and/or stirring at 500-1500 r/min in the step (2) to form an oil phase; and/or in the step (3), the emulsification process is stirring at 500-1500 r/min, the emulsification temperature is 20-50 ℃, and the emulsification time is 20-90 min; and/or, in the step (4), the curing time is 0.5-3 h, and the curing temperature is 30-70 ℃; and/or, the cross-linking agent in the step (4) is selected from one or more of dicumyl peroxide, benzoyl peroxide, di-tert-butyl peroxide, dicumyl peroxide, 2-ethyl-4-methylimidazole, 2-phenylimidazole, hexahydrophthalic anhydride, triethylenetetramine, dimethylaminopropylamine, diethylaminopropylamine, melamine, polyethylene glycol, divinylbenzene, formaldehyde and glutaraldehyde; and/or, in the step (4), the curing is carried out under the stirring of 500-1500 r/min; and/or the dosage ratio of the carboxymethyl chitosan microcapsule, the folic acid, the NHS and the EDC in the step (5) is (0.01-0.2): 0.01-0.2) by molar mass ratio of 1.
9. Use of the microcapsule according to any one of claims 1 to 6 for preparing an antitumor medical agent.
10. Use according to claim 9, characterized in that: the antitumor diagnosis and treatment agent is used for tumor embolism treatment and/or photothermal treatment and/or chemotherapy and/or disease imaging diagnosis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010930177.7A CN112121162A (en) | 2020-09-07 | 2020-09-07 | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010930177.7A CN112121162A (en) | 2020-09-07 | 2020-09-07 | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112121162A true CN112121162A (en) | 2020-12-25 |
Family
ID=73848281
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010930177.7A Pending CN112121162A (en) | 2020-09-07 | 2020-09-07 | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112121162A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101724163A (en) * | 2009-06-23 | 2010-06-09 | 河南科技大学 | Composite microsphere of chitosan derivative, preparing method and applications thereof |
| WO2014141289A1 (en) * | 2013-03-12 | 2014-09-18 | Amrita Vishwa Vidyapeetham University | Photo - chemo composition on the basis of microcapsules with a core -shell structure |
| CN104225631A (en) * | 2014-09-05 | 2014-12-24 | 中国科学院理化技术研究所 | Microcapsule with functions of magnetic resonance imaging and microwave treatment of tumors and preparation method and application thereof |
| CN110433143A (en) * | 2019-07-29 | 2019-11-12 | 同济大学 | Perfluocarbon oxygen carrying microballoon and its preparation method and application |
| CN111588853A (en) * | 2020-06-01 | 2020-08-28 | 南京林业大学 | Preparation method and application of chemotherapy-phototherapy synergistic antitumor microspheres |
-
2020
- 2020-09-07 CN CN202010930177.7A patent/CN112121162A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101724163A (en) * | 2009-06-23 | 2010-06-09 | 河南科技大学 | Composite microsphere of chitosan derivative, preparing method and applications thereof |
| WO2014141289A1 (en) * | 2013-03-12 | 2014-09-18 | Amrita Vishwa Vidyapeetham University | Photo - chemo composition on the basis of microcapsules with a core -shell structure |
| CN104225631A (en) * | 2014-09-05 | 2014-12-24 | 中国科学院理化技术研究所 | Microcapsule with functions of magnetic resonance imaging and microwave treatment of tumors and preparation method and application thereof |
| CN110433143A (en) * | 2019-07-29 | 2019-11-12 | 同济大学 | Perfluocarbon oxygen carrying microballoon and its preparation method and application |
| CN111588853A (en) * | 2020-06-01 | 2020-08-28 | 南京林业大学 | Preparation method and application of chemotherapy-phototherapy synergistic antitumor microspheres |
Non-Patent Citations (2)
| Title |
|---|
| I-CHIEH LIN ET AL: "Enhanced Targeted Delivery of Cyclodextrin-Based Supermolecules by Core–Shell Nanocapsules for Magnetothermal Chemotherapy", 《MACROMOL. BIOSCI.》 * |
| 杨栋梁 等: "羧甲基壳聚糖及其复合纳米粒子作为药物载体的应用", 《中南药学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108434462B (en) | Multifunctional nano diagnosis and treatment agent with mesoporous polydopamine loaded carbonyl manganese and preparation method and application thereof | |
| US9433800B2 (en) | Activatable particles, preparation and uses | |
| Zhang et al. | Near-infrared-triggered in situ hybrid hydrogel system for synergistic cancer therapy | |
| Wang et al. | Pullulan-coated phospholipid and Pluronic F68 complex nanoparticles for carrying IR780 and paclitaxel to treat hepatocellular carcinoma by combining photothermal therapy/photodynamic therapy and chemotherapy | |
| WO2019114066A1 (en) | Black phosphorus-based hydrogel near-infrared light-controllable drug release system and preparation method therefor | |
| CN107007835B (en) | Prussian blue-loaded targeting nano-composite and preparation method thereof | |
| Guo et al. | Emerging biocompatible nanoplatforms for the potential application in diagnosis and therapy of deep tumors | |
| CN108452303A (en) | It is a kind of to carry double medicine nanometer formulations and preparation method thereof | |
| CN107638566B (en) | Multifunctional nanocapsule integrating near-infrared fluorescence imaging and chemotherapy/photothermal therapy | |
| CN104623658A (en) | Water-soluble ferrate composite nano-particle as well as preparation method and application thereof | |
| CN104829652A (en) | Near infrared light-controlled nitrogen monoxide release nanosystem, and preparation method and application thereof | |
| AU2004206770B2 (en) | Microparticles for selectively targeted hyperthermia | |
| CN114010799B (en) | Targeting degradable nano-drug carrier for photo-thermal synergistic chemotherapy and preparation method thereof | |
| CN108912349A (en) | Polylactic acid microsphere and preparation method thereof and the application in medicament slow release | |
| CN111450269A (en) | Multifunctional ultrasonic contrast agent and preparation method thereof | |
| Wang et al. | Polyethylenimine (PEI)-modified poly (lactic-co-glycolic) acid (PLGA) nanoparticles conjugated with tumor-homing bacteria facilitate high intensity focused ultrasound-mediated tumor ablation | |
| Alvi et al. | Chitosan IR806 dye-based polyelectrolyte complex nanoparticles with mitoxantrone combination for effective chemo-photothermal therapy of metastatic triple-negative breast cancer | |
| Bao et al. | Applications of phase change materials in smart drug delivery for cancer treatment | |
| Zhou et al. | Photothermally responsive theranostic nanocomposites for near‐infrared light triggered drug release and enhanced synergism of photothermo‐chemotherapy for gastric cancer | |
| CN112546025B (en) | Preparation method of Ce6@CMCS-DSP-IPI549 anti-tumor nano-delivery system | |
| Cheng et al. | Injectable hydrogels as emerging drug-delivery platforms for tumor therapy | |
| Meng et al. | Short-wavelength aggregation-induced emission photosensitizers for solid tumor therapy: Enhanced with white-light fiber optic | |
| CN104069087B (en) | Microcapsule for microwave tumor sensitization treatment and preparation method and application thereof | |
| CN108524470A (en) | A kind of Nano medication system and its preparation method and application for treating triple negative breast cancer | |
| CN112121162A (en) | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application 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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201225 |