CN109045308B - Preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide - Google Patents
Preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide Download PDFInfo
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 37
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 37
- 150000004676 glycans Chemical class 0.000 title claims abstract description 36
- 240000000588 Hericium erinaceus Species 0.000 title claims abstract description 33
- 235000007328 Hericium erinaceus Nutrition 0.000 title claims abstract description 33
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 26
- 239000002041 carbon nanotube Substances 0.000 title claims abstract description 24
- 229910021393 carbon nanotube Inorganic materials 0.000 title claims abstract description 24
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 239000002048 multi walled nanotube Substances 0.000 claims abstract description 15
- 238000011068 loading method Methods 0.000 claims abstract description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 238000005303 weighing Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 238000009210 therapy by ultrasound Methods 0.000 claims description 6
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 6
- 239000012498 ultrapure water Substances 0.000 claims description 6
- 238000002474 experimental method Methods 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 claims description 3
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims 2
- 239000003814 drug Substances 0.000 abstract description 18
- 229940079593 drug Drugs 0.000 abstract description 13
- 238000005538 encapsulation Methods 0.000 abstract description 5
- 239000003937 drug carrier Substances 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 238000011160 research Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000011796 hollow space material Substances 0.000 description 1
- 230000008799 immune stress Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 238000007709 nanocrystallization Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
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- 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
- A61K47/6927—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 solid microparticle having no hollow or gas-filled cores
- A61K47/6929—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 solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
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- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- 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
- A61K47/56—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 macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—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 macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract
The invention provides a preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide, which is characterized in that hericium erinaceus polysaccharide is loaded on chitosan-carboxylated multi-wall carbon nano tubes, and the modified carbon nano tubes can reduce the toxicity and increase the application on organisms. High drug loading rate and encapsulation rate. The membrane-spanning capability in cells is strong, and a theoretical basis is provided for the application of the functionalized carbon nanotube on a drug carrier material.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide.
Background
Nowadays, nano-drugs are a key research direction in the field of biological medicine, and people hope that the nano-drugs can avoid the disadvantages of the existing drugs and improve the curative effect of the drugs. In the past decades, some nanocarriers such as liposomes, micelles, silica nanoparticles, carbon nanotubes, etc. have been synthesized. A large number of modern researches show that the hericium erinaceus polysaccharide has the effects of resisting oxidation, enhancing immunity, resisting aging, reducing blood sugar and the like, but the bioavailability is not high, and the loss of effective components is serious. According to the research, the hericium erinaceus polysaccharide is modified by the chitosan/carbon nano tubes, the carbon nano tubes have the structure of ultra-light weight, hollow space inside and high surface area, so that the carbon nano tubes can well encapsulate the medicine, the strongest capacity of the carbon nano tubes is the transmembrane capacity of cells, and the carbon nano tubes can become an ideal carrier in the medicine transportation process. The untreated carbon nanotubes have poor biocompatibility and potential toxicity. The chitosan has good biocompatibility, low toxicity and easy preparation. The chitosan is used for functionally modifying the carbon nano tube to improve the water solubility of the carbon nano tube, and then hericium erinaceus polysaccharide is used as a model drug to find that the hericium erinaceus polysaccharide loaded by the chitosan carbon nano tube can enhance the drug loading rate and the encapsulation rate compared with the traditional polysaccharide nanocrystallization method, thereby providing a theoretical basis for the application of the functionalized carbon nano tube on a drug carrier material.
There are two problems in the present research on drug loading of hericium erinaceus polysaccharide: 1. the effective ingredients of Chinese medicine are rarely studied on carbon nanotubes. The study adopts hericium erinaceus polysaccharide, and fills the blank in traditional Chinese medicine. 2. The drug loading after loading the drug onto the carrier is low. 3. After the vector study was successful, it was not studied subsequently.
Disclosure of Invention
The invention aims to provide a preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide comprises the following steps:
(1) preparing chitosan-carboxylated multi-walled carbon nanotubes;
1) weighing 0.2 g of chitosan CS, using 0.1 mol/L acetic acid to fix the volume to 100 mL, and carrying out ultrasonic treatment for 30 min;
2) adding 50 mg of multi-walled carbon nanotubes (MWCNTs), and performing ultrasonic treatment for 30 min;
3) uniformly dropwise adding 40 mL of 1mg/mL TPP solution, and magnetically stirring for 5 hours;
4) centrifuging the mixture at 13000 rpm for 30 min, and washing with ultrapure water for 3 times;
5) further experiments with freeze-drying to obtain chitosan-carboxylated multiwalled carbon nanotube (CS-MWCNTs) powder.
(2) Loading hericium erinaceus polysaccharide.
1) Accurately weighing 5 mg of chitosan-carboxylated multi-walled carbon nanotubes (CS-MWCNTs), dissolving in 8 mL of deionized water, and performing ultrasonic dispersion for 5 min;
2) weighing 20 mg of hericium erinaceus polysaccharide, dissolving the hericium erinaceus polysaccharide in deionized water, adding the dissolved hericium erinaceus polysaccharide into the solution obtained in the step (1), and ultrasonically mixing and dispersing for 30 min; ensuring that the chitosan-carboxylated multi-walled carbon nano-tube (CS-MWCNTs) is 0.5 mg/mL and the hericium erinaceus polysaccharide is 2.0 mg/mL;
3) centrifuging the mixture at 13000 rpm for 30 min, and washing with ultrapure water for 3 times;
4) measuring the sugar concentration of the supernatant, and freeze-drying the mixture of chitosan/carbon nano tube/hericium erinaceus polysaccharide (CS-MWCNTs-Hep) for later use.
The invention has the advantages that:
1. the modified carbon nano tube can reduce the toxicity and increase the application in organisms.
2. High drug loading rate and encapsulation rate.
3. The membrane-spanning capability in cells is strong, and a theoretical basis is provided for the application of the functionalized carbon nanotube on a drug carrier material.
Detailed Description
Example 1
A preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide comprises the following steps:
(1) preparing chitosan-carboxylated multi-walled carbon nanotubes;
1) weighing 0.2 g of CS, diluting to 100 mL with 0.1 mol/L acetic acid, and carrying out ultrasonic treatment for 30 min;
2) adding 50 mg MWCNTs, and performing ultrasonic treatment for 30 min;
3) uniformly dropwise adding 40 mL of 1mg/mL TPP solution, and magnetically stirring for 5 hours;
4) centrifuging the mixture at 13000 rpm for 30 min, and washing with ultrapure water for 3 times;
5) further experiments with freeze drying to obtain CS-MWCNTs powder.
(2) Loading hericium erinaceus polysaccharide:
1) accurately weighing 5 mg of CS-MWCNTs, dissolving in 8 mL of deionized water, and performing ultrasonic dispersion for 5 min;
2) weighing 20 mg of hericium erinaceus polysaccharide, dissolving the hericium erinaceus polysaccharide in deionized water, adding the dissolved hericium erinaceus polysaccharide into the solution obtained in the step (1), and ultrasonically mixing and dispersing for 30 min; ensuring that CS-MWCNTs is 0.5 mg/mL, and hericium erinaceus polysaccharide is 2.0 mg/mL;
3) centrifuging the mixture at 13000 rpm for 30 min, and washing with ultrapure water for 3 times;
4) measuring the sugar concentration of the supernatant, and freeze-drying the CS-MWCNTs-Hep mixture for later use.
The zeta potential of CS-MWCNTs modified hericium erinaceus polysaccharide is larger than that of the original carbon nano tube, and the polysaccharide solution is more stable. Compared with other nano-polysaccharides, the CS-MWCNTs-Hep has the characteristics of large drug loading rate and high encapsulation rate, and the drug loading rate and the encapsulation rate of the CS-MWCNTs-Hep are 41.7% and 98.60%, respectively. CS-MWCNTs-Hep has proliferation effect when acting on IPEC-J2 cells, and can reduce the content of H2O2(ii) induced apoptosis; in animal experiments, compared with hericium erinaceus polysaccharide, the protective effect of the polysaccharide on immune stress mice can be obviously enhanced, and the immunity of the mice is improved.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (1)
1. A preparation method of chitosan/carbon nano tube/hericium erinaceus polysaccharide is characterized by comprising the following steps: the method comprises the following steps:
(1) preparing chitosan-carboxylated multi-walled carbon nanotubes;
(2) loading hericium erinaceus polysaccharide;
the preparation method of the chitosan-carboxylated multi-wall carbon nanotube comprises the following steps:
(1) weighing 0.2 g of chitosan, using 0.1 mol/L acetic acid to fix the volume to 100 mL, and carrying out ultrasonic treatment for 30 min;
(2) adding 50 mg of multi-wall carbon nano-tube, and carrying out ultrasonic treatment for 30 min;
(3) uniformly dropwise adding 40 mL of 1mg/mL TPP solution, and magnetically stirring for 5 hours;
(4) centrifuging the mixture, and cleaning with ultrapure water for 3 times;
(5) freeze-drying to obtain chitosan-carboxylated multi-walled carbon nanotube powder for further experiments;
the loading steps of the hericium erinaceus polysaccharide are as follows:
(1) accurately weighing 5 mg of chitosan-carboxylated multi-walled carbon nanotube, dissolving in 8 mL of deionized water, and performing ultrasonic dispersion for 5 min;
(2) weighing 20 mg of hericium erinaceus polysaccharide, dissolving the hericium erinaceus polysaccharide in deionized water, adding the solution in the step (1), and ultrasonically mixing and dispersing for 30 min; ensuring that the chitosan-carboxylated multi-walled carbon nano-tube is 0.5 mg/mL and the hericium erinaceus polysaccharide is 2.0 mg/mL;
(3) centrifuging the mixture, and cleaning with ultrapure water for 3 times;
(4) measuring the sugar concentration of the supernatant, and freeze-drying the CS-MWCNTs-Hep mixture for later use.
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| CF01 | Termination of patent right due to non-payment of annual fee |