CN115192543B - Preparation method of lipid-loaded soluble pigment nanoparticles - Google Patents
Preparation method of lipid-loaded soluble pigment nanoparticles Download PDFInfo
- Publication number
- CN115192543B CN115192543B CN202210761679.0A CN202210761679A CN115192543B CN 115192543 B CN115192543 B CN 115192543B CN 202210761679 A CN202210761679 A CN 202210761679A CN 115192543 B CN115192543 B CN 115192543B
- Authority
- CN
- China
- Prior art keywords
- soluble pigment
- solution
- loaded
- lipid
- nanoparticle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 120
- 239000000049 pigment Substances 0.000 title claims abstract description 53
- 150000002632 lipids Chemical class 0.000 title claims abstract description 18
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 229920001690 polydopamine Polymers 0.000 claims abstract description 100
- 229920001661 Chitosan Polymers 0.000 claims abstract description 54
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 60
- 239000000243 solution Substances 0.000 claims description 43
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 42
- 239000002202 Polyethylene glycol Substances 0.000 claims description 41
- 229920001223 polyethylene glycol Polymers 0.000 claims description 41
- 238000003756 stirring Methods 0.000 claims description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 239000008367 deionised water Substances 0.000 claims description 20
- 229910021641 deionized water Inorganic materials 0.000 claims description 20
- 238000005406 washing Methods 0.000 claims description 20
- 239000011259 mixed solution Substances 0.000 claims description 16
- 238000004108 freeze drying Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 10
- CTENFNNZBMHDDG-UHFFFAOYSA-N Dopamine hydrochloride Chemical compound Cl.NCCC1=CC=C(O)C(O)=C1 CTENFNNZBMHDDG-UHFFFAOYSA-N 0.000 claims description 10
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 10
- 229960001149 dopamine hydrochloride Drugs 0.000 claims description 10
- 229920001992 poloxamer 407 Polymers 0.000 claims description 10
- 238000005303 weighing Methods 0.000 claims description 9
- 239000000839 emulsion Substances 0.000 claims description 8
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 239000002244 precipitate Substances 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 11
- 229940079593 drug Drugs 0.000 abstract description 10
- 206010028980 Neoplasm Diseases 0.000 abstract description 8
- 239000011148 porous material Substances 0.000 abstract description 7
- 239000011248 coating agent Substances 0.000 abstract description 4
- 238000000576 coating method Methods 0.000 abstract description 4
- 238000011068 loading method Methods 0.000 abstract description 4
- 238000001727 in vivo Methods 0.000 abstract description 3
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 206010013911 Dysgeusia Diseases 0.000 abstract description 2
- 230000001934 delay Effects 0.000 abstract description 2
- 230000002209 hydrophobic effect Effects 0.000 abstract description 2
- 239000002086 nanomaterial Substances 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 230000001225 therapeutic effect Effects 0.000 abstract 1
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 84
- 229940109262 curcumin Drugs 0.000 description 58
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 43
- JEVVKJMRZMXFBT-XWDZUXABSA-N Lycophyll Natural products OC/C(=C/CC/C(=C\C=C\C(=C/C=C/C(=C\C=C\C=C(/C=C/C=C(\C=C\C=C(/CC/C=C(/CO)\C)\C)/C)\C)/C)\C)/C)/C JEVVKJMRZMXFBT-XWDZUXABSA-N 0.000 description 43
- OAIJSZIZWZSQBC-GYZMGTAESA-N lycopene Chemical compound CC(C)=CCC\C(C)=C\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C=C(/C)CCC=C(C)C OAIJSZIZWZSQBC-GYZMGTAESA-N 0.000 description 43
- 239000001751 lycopene Substances 0.000 description 43
- 235000012661 lycopene Nutrition 0.000 description 43
- 229960004999 lycopene Drugs 0.000 description 43
- ZCIHMQAPACOQHT-ZGMPDRQDSA-N trans-isorenieratene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/c1c(C)ccc(C)c1C)C=CC=C(/C)C=Cc2c(C)ccc(C)c2C ZCIHMQAPACOQHT-ZGMPDRQDSA-N 0.000 description 43
- 235000012754 curcumin Nutrition 0.000 description 42
- 239000004148 curcumin Substances 0.000 description 42
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 42
- 230000015572 biosynthetic process Effects 0.000 description 19
- 238000003786 synthesis reaction Methods 0.000 description 19
- 239000002245 particle Substances 0.000 description 16
- 239000012530 fluid Substances 0.000 description 12
- 238000001179 sorption measurement Methods 0.000 description 12
- 238000009826 distribution Methods 0.000 description 8
- 230000000968 intestinal effect Effects 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical compound O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 description 6
- 230000005540 biological transmission Effects 0.000 description 6
- 239000000969 carrier Substances 0.000 description 6
- 230000002496 gastric effect Effects 0.000 description 6
- 238000000502 dialysis Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 210000005229 liver cell Anatomy 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000002336 sorption--desorption measurement Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 238000007605 air drying Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000012876 carrier material Substances 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000013270 controlled release Methods 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 210000004051 gastric juice Anatomy 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 210000003494 hepatocyte Anatomy 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 238000001000 micrograph Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 244000241235 Citrullus lanatus Species 0.000 description 1
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 description 1
- 244000164480 Curcuma aromatica Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010019851 Hepatotoxicity Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 238000006845 Michael addition reaction Methods 0.000 description 1
- 208000027089 Parkinsonian disease Diseases 0.000 description 1
- 206010034010 Parkinsonism Diseases 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 241000234299 Zingiberaceae Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 229920006317 cationic polymer Polymers 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000009982 effect on human Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000010579 first pass effect Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 230000036449 good health Effects 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 230000007686 hepatotoxicity Effects 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- -1 polyphenol compound Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Images
Classifications
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5161—Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
-
- 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
- A61K31/01—Hydrocarbons
-
- 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
- A61K31/12—Ketones
-
- 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
- A61K47/56—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
- A61K47/59—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 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—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 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Nanotechnology (AREA)
- Optics & Photonics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to the technical field of nano materials, and discloses a preparation method of lipid-loaded pigment nanoparticles. The carrier mesoporous polydopamine has higher specific surface area and nano-pore structure, can adsorb hydrophobic drugs, and can greatly improve the loading efficiency of fat-soluble pigments. The polydopamine shell layer taking the modified chitosan as the coating has pH sensitivity and gating function, effectively responds to the pH value of a tumor part, improves the concentration of effective liposoluble pigment in tumor cells, delays the effective acting time of the liposoluble pigment, and improves the bad taste of the liposoluble pigment; can improve bioavailability and in vivo and in vitro stability of liposoluble pigment, and improve tumor therapeutic effect.
Description
Description of the division
The invention discloses a divisional application of a lipid-loaded soluble pigment nanoparticle with the application number of 202011631054X and the name of 'a preparation method thereof', wherein the application date is 12/31/2020.
Technical Field
The invention relates to the technical field of nano materials, in particular to a preparation method of lipid-loaded soluble pigment nanoparticles.
Background
Curcumin is a natural active ingredient extracted from the root of curcuma species of the family Zingiberaceae, belongs to a polyphenol compound, has wide pharmacological effects of reducing blood fat, resisting tumor, resisting inflammation, promoting bile flow, resisting oxidation, resisting hepatotoxicity, resisting rheumatism, inhibiting bacteria and the like, has low toxicity and good clinical application potential, and becomes a hotspot for domestic and foreign research. Lycopene is a carotenoid extracted from plants such as tomatoes, watermelons and the like, belongs to a fat-soluble natural red pigment, belongs to hydrocarbon, has strong antioxidant function, has the effects of protecting cardiac and cerebral vessels, enhancing immunity, resisting tumors and the like, has the effects of preventing or treating neurodegenerative diseases such as parkinsonism, epilepsy, depression and the like, is basically nontoxic in dosage range, has good health care value, and has become a hotspot of domestic and foreign research. Curcumin and lycopene both belong to liposoluble pigments, which are insoluble in water, are not easy to be absorbed by oral administration, have serious liver first pass effect, have relatively high metabolism and clearance rate in vivo, have relatively low bioavailability and poor stability, and are easy to decompose by visible light, so that a proper drug delivery system is required to be prepared to solve the problems, improve the water solubility and stability of the liposoluble pigments, prevent the drugs from being hydrolyzed and oxidized to be inactivated after entering organisms, and prolong the in vivo release time of the drugs. Polydopamine (PDA) is a main component of natural biological pigment-melanin, can be obtained through oxidation self-polymerization reaction of dopamine, has good stability, biodegradability, biocompatibility and photo-thermal conversion characteristics, and is an ideal carrier material. The polydopamine has adhesiveness and can be coated on the surfaces of various materials. Polydopamine is also pH sensitive and depolymerizes in the slightly acidic environment of tumors. The mesoporous polydopamine nanoparticle (MPDA) can be prepared by a soft template method, and has a pore structure, a high specific surface area, high drug loading efficiency and good photo-thermal conversion performance. Chitosan is a cationic polymer composed of glucosamine, has good biocompatibility, low toxicity and biodegradability, has intestinal mucosa adhesion property, and is favorable for oral absorption of medicines as a medicine auxiliary material. The polyethylene glycol modification is carried out on the chitosan, so that the adsorption effect of plasma protein on the chitosan coated mesoporous polydopamine nanoparticles can be reduced, the ingestion of macrophages on the chitosan coated mesoporous polydopamine nanoparticles is reduced, the process that the drug-carrying nanoparticles are removed from the plasma is delayed, and the passive targeting function of the chitosan mesoporous polydopamine nanoparticles is further improved through the enhanced permeation and retention effect.
Disclosure of Invention
The invention aims to: aiming at the problems existing in the prior art, the invention provides the preparation method of the lipid-loaded soluble pigment nanoparticle, which can improve the bioavailability of the lipid-soluble pigment, improve the water solubility and stability of the lipid-soluble pigment and improve the bioavailability of the lipid-soluble pigment, and is expected to achieve the purposes of tumor permeation and directional slow release.
The technical scheme is as follows: the invention provides lipid-loaded soluble pigment nanoparticles, which comprise the following components in percentage by mass: 8-13: 5-10 mesoporous polydopamine nanoparticles, a fat-soluble pigment and polyethylene glycol modified chitosan, wherein the mesoporous polydopamine nanoparticles are used as a carrier, the fat-soluble pigment is adsorbed by physical and chemical adsorption, and the polyethylene glycol modified chitosan is wrapped on the outermost layer.
Preferably, the fat-soluble pigment is curcumin or lycopene.
The invention also provides a preparation method of the lipid-loaded soluble pigment nanoparticle, which comprises the following steps: (1) Adding dopamine hydrochloride and Pluronic F127 into an ethanol water solution, stirring at room temperature, then dropwise adding TMB to form white emulsion, then adding an ammonia water solution, stirring, centrifuging, ultrasonically washing the precipitate with ethanol and water for several times, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking the mesoporous polydopamine nanoparticles as MPDA; the weight-volume ratio of the dopamine hydrochloride to the Pluronic F127 to the TMB to the ammonia solution is 0.3-0.5 g: 1.0-1.2 g: 0.6-1.0 mL: 4.0-5.0 mL; (2) Adding the mesoporous polydopamine nanoparticle obtained in the step (1) and fat-soluble pigment powder into anhydrous DMSO, stirring at room temperature for reaction, centrifuging, flushing with a mixed solution of DMSO and deionized water and flushing with deionized water for several times to obtain the lipid-soluble pigment nanoparticle; the mass ratio of the mesoporous polydopamine nanoparticle to the fat-soluble pigment powder is 9-11: 1, a step of; (3) Weighing a certain amount of chitosan and polyethylene glycol, dissolving in a dilute acetic acid solution, uniformly mixing, and stirring at room temperature overnight to obtain a polyethylene glycol modified chitosan solution; (4) Dissolving the lipid-loaded soluble pigment nanoparticle obtained in the step (2) in acetic acid aqueous solution, dropwise adding the polyethylene glycol modified chitosan solution obtained in the step (3), stirring at room temperature, centrifuging, and freeze-drying to obtain the lipid-loaded soluble pigment nanoparticle.
Preferably, in the step (1), in the ethanol aqueous solution, the volume ratio of ethanol to water is 1:1.
preferably, in the step (2), in the DMSO-deionized water mixed solution, the volume ratio of DMSO to deionized water is 3 to 4: 7-8.
Preferably, in the step (3), the mass ratio of chitosan to polyethylene glycol is 1:0.2 to 0.3.
Preferably, in the step (3), the mass fraction of the dilute acetic acid aqueous solution is 1-2%.
Preferably, in the step (4), the mass fraction of the acetic acid aqueous solution is 0.5-1%.
Preferably, in the step (4), the freeze-drying temperature is-40 to-70 ℃ and the freeze-drying time is 12-24 hours.
The beneficial effects are that: compared with the prior art, the invention has the following beneficial effects:
(1) According to the invention, the drug-loaded mesoporous polydopamine nanoparticle which takes the modified chitosan as a coating and can improve the bioavailability of the fat-soluble pigment is constructed by taking polydopamine as a base material and through synthesis of mesoporous polydopamine nanoparticle, loading of fat-soluble pigment and coating of modified chitosan molecules, so that the water solubility and stability of the fat-soluble pigment are improved, the bioavailability of the fat-soluble pigment is improved, and the drug-loaded nanoparticle is expected to achieve the purposes of tumor permeation and directional slow release.
(2) The carrier Mesoporous Polydopamine (MPDA) has higher specific surface area and nano pore canal structure, has strong adsorption capacity, can adsorb hydrophobic drug fat-soluble pigment on the surface and in the pore canal of the mesoporous polydopamine, can generate pi-pi electron transition, and forms carbonyl bond with fat-soluble pigment molecules (the fat-soluble pigment is curcumin) or generates michael addition reaction (the fat-soluble pigment is lycopene), and can greatly improve the loading efficiency of the polydopamine on the fat-soluble pigment through physical adsorption and chemical adsorption combination.
(3) The lipid-loaded soluble pigment nanoparticle provided by the invention has the functions of pH responsiveness and gating in the release of the lipid-soluble pigment, can effectively respond to the pH value of a tumor part, improves the effective concentration of the lipid-soluble pigment in a tumor cell, delays the effective acting time of the lipid-soluble pigment, can improve the bad taste of the lipid-soluble pigment, and avoids bitter components of some lipid-soluble pigments from being dissolved in the mouth.
(4) The modified chitosan can be absorbed and utilized by human body, has good biocompatibility, can be biodegraded, and the chitosan oligosaccharide produced in the degradation process is not accumulated in the body, has almost no immunogenicity, and has good water solubility, and the surface electrical property of the mesoporous polydopamine nano-carrier is changed into positive electricity through chitosan modification, so that the adhesiveness of the mesoporous polydopamine carrier to tumor cells is increased. The chitosan can be adsorbed in intestinal tracts, and can be discharged out of the body in a delayed manner, so that more fat-soluble pigments are absorbed by human bodies, the bioavailability is improved, and the particle storage stability can be improved by wrapping the chitosan on the surfaces.
(5) The mesoporous polydopamine carrier constructed by the invention is safe, nontoxic, simple in preparation, single in component, capable of improving the stability of the fat-soluble pigment and convenient for storage.
Drawings
FIG. 1 is a graph showing a particle size distribution of mesoporous polydopamine carrier and mesoporous polydopamine curcumin-loaded nanoparticles;
FIG. 2 is a transmission electron microscope image of mesoporous polydopamine carrier and mesoporous polydopamine curcumin-loaded nanoparticles;
FIG. 3 is a graph of nitrogen adsorption/desorption for mesoporous polydopamine MPDA;
fig. 4 biosafety inspection of normal human hepatocytes LO2 with blank vector;
FIG. 5 is a graph showing the slow release of mesoporous polydopamine curcumin-loaded nanoparticles in simulated gastric fluid and intestinal fluid;
FIG. 6 is a graph showing the distribution of the particle size of mesoporous polydopamine carrier and mesoporous polydopamine lycopene-loaded nanoparticles;
FIG. 7 is a transmission electron microscope image of mesoporous polydopamine carrier and mesoporous polydopamine lycopene nanoparticle;
FIG. 8 is a graph of the photostability of lycopene-loaded mesoporous polydopamine nanoparticles;
figure 9 is a graph showing the slow release of mesoporous polydopamine-loaded lycopene nanoparticles in simulated gastric fluid and intestinal fluid.
FIG. 10 cytotoxicity of mesoporous polydopamine nanoparticles on liver cancer cells HepG 2.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings.
Embodiment 1:
the embodiment provides a preparation method of mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR, which is implemented by the following steps:
step 1, synthesis of MPDA
Adding 0.4. 0.4 g dopamine hydrochloride and 0.8 g Pluronic F127 to the mixed solution of ethanol water (1:1, v/v), stirring at room temperature, and then dropwise adding 0.8 mL TMB to form a white emulsion; adding 4.0 mL ammonia water solution, stirring at 50deg.C for 30 min, centrifuging, ultrasonically washing the precipitate with ethanol and water for 3 times, centrifuging to obtain mesoporous polydopamine nanoparticle, and marking as MPDA;
The mesoporous polydopamine nanoparticle and curcumin powder are mixed according to the mass ratio of 8:1 into anhydrous DMSO, stirring at room temperature for reaction 24 h, centrifuging, washing with mixed solution of DMSO and deionized water (3:7, v/v) for one time and washing with deionized water for 3 times to obtain curcumin-loaded nanoparticle labeled MPDA@CUR;
step 3, synthesis of PEG-CS@MPDA@CUR
Weighing 1 g chitosan and 0.2 g polyethylene glycol, dissolving in 1% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving the curcumin-loaded nanoparticle in 100 mL of 0.5% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-60 ℃ to 16 h to obtain the mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR.
The prepared mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR comprises the following components in percentage by mass: 8:5, mesoporous polydopamine nanoparticles, curcumin and polyethylene glycol modified chitosan are taken as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of curcumin.
Embodiment 2:
the preparation method of the mesoporous polydopamine curcumin-loaded nanoparticle is implemented by the following steps:
step 1, synthesis of MPDA
Adding 0.5 g dopamine hydrochloride and 1.2 g Pluronic F127 into a mixed solution of ethanol water (1:1, v/v), stirring at room temperature, then dropwise adding 1.0 mL TMB to form white emulsion, adding 4.0 mL ammonia water solution, stirring at 50 ℃ for 40 min, centrifuging, ultrasonically washing the precipitate with ethanol and water for 5 times, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking the mesoporous polydopamine nanoparticles as MPDA;
The mesoporous polydopamine nanoparticle and curcumin powder are mixed according to the mass ratio of 9:1 into anhydrous DMSO, stirring at room temperature to react for 12 h, centrifuging, washing with mixed solution of DMSO and deionized water (4:7, v/v) for one time and washing with deionized water for 5 times to obtain curcumin-loaded nanoparticle, and marking as MPDA@CUR;
step 3, synthesis of PEG-CS@MPDA@CUR
Weighing 1 g chitosan and 0.3 g polyethylene glycol, dissolving in 2% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving the curcumin-loaded nanoparticle in 100 mL of 0.5% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-40 ℃ for 24 h to obtain the mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR.
The prepared mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR comprises the following components in percentage by mass: 9:7, mesoporous polydopamine nanoparticles, curcumin and polyethylene glycol modified chitosan are taken as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of curcumin.
Embodiment 3:
the preparation method of the mesoporous polydopamine curcumin-loaded nanoparticle is implemented by the following steps:
step 1, synthesis of MPDA
Adding 0.3 g dopamine hydrochloride and 0.5 g Pluronic F127 into a mixed solution of ethanol water (1:1, v/v), stirring at room temperature, then dropwise adding 0.8 mL TMB to form white emulsion, adding 3 mL ammonia water solution, stirring at 40 ℃ for 30 min, centrifuging, washing the precipitate with ethanol and water for 5 times by ultrasonic, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking as MPDA;
Adding mesoporous polydopamine nanoparticles and curcumin powder into anhydrous DMSO according to the mass ratio of 10:1, stirring at room temperature for reaction 24 h, centrifuging, washing once with a mixed solution of DMSO and deionized water (4:9, v/v) and washing 5 times with deionized water to obtain curcumin-loaded nanoparticles, wherein the curcumin-loaded nanoparticles are marked as MPDA@CUR;
step 3, synthesis of PEG-CS@MPDA@CUR
Weighing 1 g chitosan and 0.2 g polyethylene glycol, dissolving in 1% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving the curcumin-loaded nanoparticle in 100 mL of 0.5% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-40 ℃ for 24 h to obtain the mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR.
The prepared mesoporous polydopamine curcumin-loaded nanoparticle PEG-CS@MPDA@CUR comprises the following components in percentage by mass: 9:6, mesoporous polydopamine nanoparticles, curcumin and polyethylene glycol modified chitosan are taken as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of curcumin.
The particle size distribution of the mesoporous polydopamine carrier and the mesoporous polydopamine-loaded curcumin nanoparticles of embodiments 1 to 3 was analyzed using a malvern laser particle sizer. The mesoporous polydopamine carrier and the mesoporous polydopamine curcumin-loaded nanoparticle are dispersed in water, and the particle size distribution is measured, and the hydrodynamic diameters of the mesoporous polydopamine carrier and the mesoporous polydopamine curcumin-loaded nanoparticle are 125+/-10 nm and 135+/-10 nm respectively as shown in figure 1.
The morphology of the mesoporous polydopamine carrier and the mesoporous polydopamine-loaded curcumin nanoparticles of embodiments 1 to 3 was observed by Transmission Electron Microscopy (TEM): and (3) taking 10 mu L of the solution, dripping the solution on the surface carbon-coated copper mesh, and naturally air-drying at room temperature. And under the condition of 200KV voltage, observing the morphology, the particle size and the dispersion condition of the nano particles by a transmission electron microscope. The carrier transmission electron microscope picture is shown in figure 2, the obtained MPDA has a narrow particle size distribution range, uniform particle size and obvious pore channel structure on the surface.
MPDA nitrogen adsorption/desorption curve determination: taking a dried 80 mg MPDA sample, measuring a nitrogen adsorption/desorption curve by an instrument, and calculating the specific surface area of the prepared MPDA nano particle to be 63.6510 m per gram by using a BJH method as shown in figure 3.
The growth inhibition effect of the blank vector on human normal hepatocytes LO2 was examined by MTT method. The normal human liver cell LO2 is used, 200 mu L/hole of blank carrier solution with different concentrations is added into an experimental group, 200 mu L of culture solution is added into a control group, and the relative cell viability is taken as an investigation index under two pH conditions to investigate the cell viability of the normal human liver cell LO2 under different concentration conditions. As shown in FIG. 4, when the concentration of the empty nanoparticle reaches 1000 mug/mL, the survival rate of the human normal liver cell LO2 is over 80%, which indicates that the carrier material has good biocompatibility within the concentration of 0.98-1000 mug/mL.
The release conditions of the mesoporous polydopamine curcumin-loaded nanoparticles in the simulated gastric fluid and the simulated intestinal fluid in the embodiments 1 to 3 are examined by a dialysis bag method. Placing the 1 mL mesoporous polydopamine curcumin-loaded nanoparticle suspension in a dialysis bag, shaking at constant temperature at 37 ℃ with release medium being simulated artificial gastric juice and artificial intestinal juice, sampling at different time points, and drawing an accumulated drug release curve. The experimental result is shown in fig. 5, and it can be seen from the graph that the release rate of the mesoporous polydopamine curcumin-loaded nanoparticle in simulated gastric fluid is higher than that in simulated intestinal fluid, the cumulative release rate is more than 80%, and the release is complete. The mesoporous polydopamine curcumin-loaded nanoparticle is slowly released from the beginning of an experiment and is gradually stable along with the time, so that the mesoporous polydopamine curcumin-loaded nanoparticle has obvious effect in the aspect of curcumin controlled release.
Embodiment 4:
the embodiment provides a preparation method of mesoporous polydopamine supported lycopene nanoparticle PEG-CS@MPDA@LYC, which is implemented by the following steps:
step 1, synthesis of MPDA
Adding 0.4. 0.4 g dopamine hydrochloride and 0.9 g Pluronic F127 to the mixed solution of ethanol water (1:1, v/v), stirring at room temperature, and then dropwise adding 1.0 mL TMB to form a white emulsion; adding 4.5 mL ammonia water solution, stirring at 50deg.C for 30 min, centrifuging, ultrasonically washing the precipitate with ethanol and water for 3 times, centrifuging to obtain mesoporous polydopamine nanoparticle, and marking as MPDA;
The mesoporous polydopamine nanoparticle and lycopene powder are mixed according to the mass ratio of 9:1 into anhydrous DMSO, stirring at room temperature for reaction 24 h, centrifuging, washing with mixed solution of DMSO and deionized water (4:7, v/v) for one time and washing with deionized water for 3 times to obtain lycopene-loaded nanoparticle, and marking as MPDA@LYC;
step 3, synthesis of PEG-CS@MPDA@LYC
Weighing 1 g chitosan and 0.25 g polyethylene glycol, dissolving in 2% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving lycopene-carrying nanoparticles in 100 mL of 0.5% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-40 ℃ for 24 h to obtain mesoporous polydopamine-carrying nanoparticles PEG-CS@MPDA@LYC.
The prepared mesoporous polydopamine lycopene-loaded nanoparticle PEG-CS@MPDA@LYC comprises the following components in percentage by mass: 9:6, mesoporous polydopamine nanoparticles, lycopene and polyethylene glycol modified chitosan are used as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of lycopene.
Embodiment 5:
the preparation method of the mesoporous polydopamine lycopene-loaded nanoparticle is specifically implemented according to the following steps:
step 1, synthesis of MPDA
Adding 0.5 g dopamine hydrochloride and 1.0 g Pluronic F127 into a mixed solution of ethanol water (1:1, v/v), stirring at room temperature, then dropwise adding 0.8 mL TMB to form white emulsion, adding 4.0 mL ammonia water solution, stirring at 50 ℃ for 40 min, centrifuging, ultrasonically washing the precipitate with ethanol and water for 5 times, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking as MPDA;
The mesoporous polydopamine nanoparticle and lycopene powder are mixed according to the mass ratio of 10:1 into anhydrous DMSO, stirring at room temperature to react for 12 h, centrifuging, washing with mixed solution of DMSO and deionized water (3:7, v/v) for one time and washing with deionized water for 5 times to obtain lycopene-carrying nanoparticle, and marking as MPDA@LYC;
step 3, synthesis of PEG-CS@MPDA@LYC
Weighing 1 g chitosan and 0.2 g polyethylene glycol, dissolving in 1.5% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving lycopene-carrying nanoparticles in 100 mL of 1.0% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-40 ℃ for 24 h to obtain mesoporous polydopamine-carrying nanoparticles PEG-CS@MPDA@LYC.
The prepared mesoporous polydopamine lycopene-loaded nanoparticle PEG-CS@MPDA@LYC comprises the following components in percentage by mass: 11: and 8, mesoporous polydopamine nanoparticles, lycopene and polyethylene glycol modified chitosan are taken as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of lycopene.
Embodiment 6:
the preparation method of the mesoporous polydopamine lycopene-loaded nanoparticle is specifically implemented according to the following steps:
step 1, synthesis of MPDA
Adding 0.4 g dopamine hydrochloride and 1.2 g Pluronic F127 into a mixed solution of ethanol water (1:1, v/v), stirring at room temperature, then dropwise adding 0.8 mL TMB to form white emulsion, adding 4 mL ammonia water solution, stirring at 40 ℃ for 30 min, centrifuging, ultrasonically washing the precipitate with ethanol and water for 5 times, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking the mesoporous polydopamine nanoparticles as MPDA;
Adding mesoporous polydopamine nanoparticles and lycopene powder into anhydrous DMSO according to the mass ratio of 11:1, stirring at room temperature to react for 24 h, centrifuging, washing once with a mixed solution of DMSO and deionized water (3:8, v/v) and washing 5 times with deionized water to obtain lycopene-carrying nanoparticles, wherein the lycopene-carrying nanoparticles are marked as MPDA@LYC;
step 3, synthesis of PEG-CS@MPDA@LYC
Weighing 1 g chitosan and 0.3 g polyethylene glycol, dissolving in 2% dilute acetic acid solution, mixing uniformly, and stirring overnight at room temperature to obtain polyethylene glycol modified chitosan solution; dissolving lycopene-carrying nanoparticles in 100 mL of 0.5% acetic acid water, dropwise adding 20 mL polyethylene glycol modified chitosan solution, stirring at room temperature, centrifuging, and freeze-drying at-40 ℃ for 24 h to obtain mesoporous polydopamine-carrying nanoparticles PEG-CS@MPDA@LYC.
The prepared mesoporous polydopamine lycopene-loaded nanoparticle PEG-CS@MPDA@LYC comprises the following components in percentage by mass: 11:7, mesoporous polydopamine nanoparticles, lycopene and polyethylene glycol modified chitosan are used as carriers, and the polyethylene glycol modified chitosan is wrapped on the outermost layer through physical and chemical adsorption of lycopene.
The particle size distribution of the mesoporous polydopamine carrier and the mesoporous polydopamine-loaded lycopene nanoparticles of embodiments 4 to 6 was analyzed by using a malvern laser particle sizer. The mesoporous polydopamine carrier and the mesoporous polydopamine lycopene nanoparticle are dispersed in water, and the particle size distribution is measured, and as shown in figure 6, the hydrodynamic diameters are 125+/-10 nm and 140+/-10 nm respectively.
The morphology of the mesoporous polydopamine carrier and the mesoporous polydopamine-loaded lycopene nanoparticle according to embodiments 4 to 6 was observed by Transmission Electron Microscopy (TEM): and (3) taking 10 mu L of the solution, dripping the solution on the surface carbon-coated copper mesh, and naturally air-drying at room temperature. And under the condition of 200KV voltage, observing the morphology, the particle size and the dispersion condition of the nano particles by a transmission electron microscope. The carrier transmission electron microscope picture is shown in fig. 7a, and the obtained MPDA has a narrow particle size distribution range, uniform particle size and obvious pore channel structure on the surface. As shown in figure 7b, the mesoporous polydopamine lycopene-loaded nanoparticle has uniform particle size and spherical shape, and regularly distributed pore channels become blurred due to adsorption of lycopene on the surface and modification of chitosan.
Placing a certain amount of lycopene-carrying nanoparticle and lycopene powder in indoor scattering light environment, spreading, performing full light irradiation to make light energy contact with each surface of lycopene, weighing a certain amount of powder at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h and 12 h respectively, adding 10 mL DMSO, dissolving thoroughly, performing ultrasound for 10 min, repeating for 3 times to obtain average value, and measuring content by ultraviolet spectrophotometry. As shown in figure 8, the stability of the lycopene is obviously improved by preparing the nanocapsules, and the lycopene content after 12 h is improved from 80.75% before the original non-adsorbed embedding to about 98.72%.
The release conditions of the mesoporous polydopamine-loaded lycopene nanoparticles in the simulated gastric fluid and the simulated intestinal fluid in the embodiments 4 to 6 are examined by a dialysis bag method. Placing 1 mL mesoporous polydopamine lycopene nanoparticle suspension in a dialysis bag, shaking at constant temperature at 37deg.C with release medium simulating artificial gastric juice and artificial intestinal juice, sampling at different time points, and drawing accumulated drug release curve. The experimental result is shown in fig. 9, and it can be seen from the graph that the release rate of the mesoporous polydopamine lycopene-loaded nanoparticle in simulated gastric fluid is higher than that in simulated intestinal fluid, the cumulative release rate is also higher than 80%, and the release is more complete. And the slow release is carried out from the beginning of the experiment, and the slow release is gradually stable along with the time, which proves that the mesoporous polydopamine lycopene-loaded nanoparticle has obvious effect in the aspect of lycopene controlled release.
The toxic effect of free lycopene, PEG-CS@MPDA@LYC on human hepatoma cells HepG2 was examined by MTT assay. The results are shown in FIG. 10, and lycopene shows obvious dose-dependent inhibition effect on human hepatoma cell HepG2 under two pH conditions. This enhanced antitumor effect of lycopene after being supported on a carrier may be due to the excellent antiproliferative activity of the chelated lycopene and the synergistic antitumor effect of lycopene and the surface modified chitosan coating.
The foregoing embodiments are merely illustrative of the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the present invention and to implement the same, not to limit the scope of the present invention. All equivalent changes or modifications made according to the spirit of the present invention should be included in the scope of the present invention.
Claims (7)
1. The preparation method of the lipid-loaded soluble pigment nanoparticle is characterized by comprising the following steps of:
(1) Adding dopamine hydrochloride and Pluronic F127 into an ethanol water solution, stirring at room temperature, then dropwise adding TMB to form white emulsion, then adding an ammonia water solution, stirring, centrifuging, ultrasonically washing the precipitate with ethanol and water for several times, centrifuging to obtain mesoporous polydopamine nanoparticles, and marking the mesoporous polydopamine nanoparticles as MPDA;
the weight-volume ratio of the dopamine hydrochloride to the Pluronic F127 to the TMB to the ammonia solution is 0.3-0.5 g: 1.0-1.2 g: 0.6-1.0 mL: 4.0-5.0 mL;
(2) Adding the mesoporous polydopamine nanoparticle obtained in the step (1) and fat-soluble pigment powder into anhydrous DMSO, stirring at room temperature for reaction, centrifuging, flushing with a mixed solution of DMSO and deionized water and flushing with deionized water for several times to obtain the lipid-soluble pigment nanoparticle;
the mass ratio of the mesoporous polydopamine nanoparticle to the fat-soluble pigment powder is 9-11: 1, a step of;
(3) Weighing a certain amount of chitosan and polyethylene glycol, dissolving in a dilute acetic acid solution, uniformly mixing, and stirring at room temperature overnight to obtain a polyethylene glycol modified chitosan solution;
(4) Dissolving the lipid-loaded soluble pigment nanoparticle obtained in the step (2) in acetic acid aqueous solution, dropwise adding the polyethylene glycol modified chitosan solution obtained in the step (3), stirring at room temperature, centrifuging, and freeze-drying to obtain the lipid-loaded soluble pigment nanoparticle.
2. The method of claim 1, wherein in the step (1), the volume ratio of ethanol to water in the aqueous ethanol solution is 1:1.
3. the method for preparing lipid-loaded soluble pigment nanoparticles according to claim 1, wherein in the step (2), the volume ratio of DMSO to deionized water in the mixed solution of DMSO and deionized water is 3-4: 7-8.
4. The method for preparing lipid-loaded soluble pigment nanoparticles according to claim 1, wherein in the step (3), the mass ratio of chitosan to polyethylene glycol is 1:0.2-0.3.
5. The method of claim 4, wherein in the step (3), the mass fraction of the dilute aqueous acetic acid solution is 1-2%.
6. The method for preparing lipid-loaded soluble pigment nanoparticles according to claim 1, wherein in the step (4), the mass fraction of the aqueous acetic acid solution is 0.5-1%.
7. The method for preparing lipid-loaded soluble pigment nanoparticles according to any one of claims 1 to 6, wherein in the step (4), the freeze-drying temperature is-40 to-70 ℃ and the freeze-drying time is 12 to 24 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210761679.0A CN115192543B (en) | 2020-12-31 | 2020-12-31 | Preparation method of lipid-loaded soluble pigment nanoparticles |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210761679.0A CN115192543B (en) | 2020-12-31 | 2020-12-31 | Preparation method of lipid-loaded soluble pigment nanoparticles |
| CN202011631054.XA CN112546027B (en) | 2020-12-31 | 2020-12-31 | Fat-soluble pigment-loaded nanoparticle and preparation method thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011631054.XA Division CN112546027B (en) | 2020-12-31 | 2020-12-31 | Fat-soluble pigment-loaded nanoparticle and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115192543A CN115192543A (en) | 2022-10-18 |
| CN115192543B true CN115192543B (en) | 2023-06-30 |
Family
ID=75035050
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210761679.0A Active CN115192543B (en) | 2020-12-31 | 2020-12-31 | Preparation method of lipid-loaded soluble pigment nanoparticles |
| CN202011631054.XA Active CN112546027B (en) | 2020-12-31 | 2020-12-31 | Fat-soluble pigment-loaded nanoparticle and preparation method thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011631054.XA Active CN112546027B (en) | 2020-12-31 | 2020-12-31 | Fat-soluble pigment-loaded nanoparticle and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN115192543B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114225098B (en) * | 2021-11-24 | 2023-04-11 | 中国科学院理化技术研究所 | Medical dressing with antibacterial and wound healing promoting functions |
| CN114225048B (en) * | 2021-12-30 | 2023-05-09 | 上海交通大学医学院附属第九人民医院 | Mesoporous dopamine drug delivery system and application thereof in treating knee osteoarthritis |
| CN114887117B (en) * | 2022-04-09 | 2023-10-13 | 中国医科大学附属第一医院 | Drug-loaded nanoparticle and vascular stent for reducing restenosis rate in blood vessel after operation |
| CN117224708B (en) * | 2023-11-10 | 2024-02-06 | 烟台大学 | Illicit Mo Nami preparation and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107312205A (en) * | 2017-06-15 | 2017-11-03 | 淮阴工学院 | The preparation and its application of recessed native loading chitosan cross linked ciclodextrines compound |
| CN109276721A (en) * | 2018-09-19 | 2019-01-29 | 中山大学 | It is a kind of to target mesoporous poly-dopamine multifunctional nano diagnosis and treatment agent and the preparation method and application thereof |
| CN110201163A (en) * | 2019-06-17 | 2019-09-06 | 重庆医科大学 | A kind of load medicine mesoporous TiO 2 nanoparticle of hyaluronic acid and poly-dopamine modification |
| CN111110652A (en) * | 2020-01-06 | 2020-05-08 | 重庆医科大学 | Drug-loaded hyaluronic acid polydopamine-coated mesoporous polydopamine nanoparticle and preparation method thereof |
| WO2020172618A2 (en) * | 2019-02-21 | 2020-08-27 | Bambu Vault Llc | Remotely triggered therapy |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160346208A1 (en) * | 2015-05-28 | 2016-12-01 | University Of South Carolina | Dual responsive brain targeted nanoparticles and their applications |
| CN105963702A (en) * | 2016-04-27 | 2016-09-28 | 湖北大学 | Drug delivery controlled-release system having tumor induced targeting capacity and preparation method of drug delivery controlled-release system |
| CN106806343B (en) * | 2017-02-17 | 2020-10-13 | 清华大学深圳研究生院 | Folic acid and polydopamine modified tumor targeting mesoporous silica nanoparticle and preparation method and application thereof |
| CN106806344B (en) * | 2017-02-17 | 2020-10-30 | 清华大学深圳研究生院 | Polydopamine and polyethylene glycol vitamin E succinate modified mesoporous silica nanoparticle and preparation method and application thereof |
| CN108653754B (en) * | 2018-07-06 | 2021-05-18 | 重庆医科大学 | Hyaluronic acid targeted polydopamine coated phase-change type liquid fluorocarbon nano ultrasonic contrast agent |
| CN110204711B (en) * | 2019-05-31 | 2021-11-30 | 中国药科大学 | Preparation method and application of pomegranate-like structure hollow mesoporous molecularly imprinted polydopamine nanoparticle adsorbent |
| CN110693851B (en) * | 2019-10-24 | 2022-03-25 | 西安医学院 | Mesoporous silica drug-loaded nanoparticle and preparation method and application thereof |
| CN110755613A (en) * | 2019-10-24 | 2020-02-07 | 暨南大学 | Preparation and application of light-triggered erythrocyte membrane-coated NO nano bionic donor material |
| CN111671914B (en) * | 2020-06-29 | 2023-05-26 | 广东省医疗器械研究所 | Near infrared light response nanoparticle and controlled release system |
-
2020
- 2020-12-31 CN CN202210761679.0A patent/CN115192543B/en active Active
- 2020-12-31 CN CN202011631054.XA patent/CN112546027B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107312205A (en) * | 2017-06-15 | 2017-11-03 | 淮阴工学院 | The preparation and its application of recessed native loading chitosan cross linked ciclodextrines compound |
| CN109276721A (en) * | 2018-09-19 | 2019-01-29 | 中山大学 | It is a kind of to target mesoporous poly-dopamine multifunctional nano diagnosis and treatment agent and the preparation method and application thereof |
| WO2020172618A2 (en) * | 2019-02-21 | 2020-08-27 | Bambu Vault Llc | Remotely triggered therapy |
| CN110201163A (en) * | 2019-06-17 | 2019-09-06 | 重庆医科大学 | A kind of load medicine mesoporous TiO 2 nanoparticle of hyaluronic acid and poly-dopamine modification |
| CN111110652A (en) * | 2020-01-06 | 2020-05-08 | 重庆医科大学 | Drug-loaded hyaluronic acid polydopamine-coated mesoporous polydopamine nanoparticle and preparation method thereof |
Non-Patent Citations (7)
| Title |
|---|
| Biological Evaluation of Black Chokeberry Extract Free and Embedded in Two Mesoporous Silica-Type Matrices;Buda V等;PHARMACEUTICS;第21卷(第9期);1-10 * |
| Highly Efficient Regioselective Decanoylation of Hyperoside Using Nanobiocatalyst of Fe3O4@PDA-Thermomyces lanuginosusLipase: Insights of Kinetics and Stability Evaluation;Bi YH，等;FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY;第8卷;1-8 * |
| Mesoporous Silica Nanoparticles (MCM-41) Coated PEGylated Chitosan as a pH-Responsive Nanocarrier for Triggered Release of Erythromycin;Ali Pourjavadi，等;International Journal of Polymeric Materials and Polymeric Biomaterials;第63卷;692-693 * |
| Mitochondria-Targeting Polydopamine Nanoparticles To Deliver Doxorubicin for Overcoming Drug Resistance;Li WQ，等;ACS APPLIED MATERIALS & INTERFACES;第9卷(第20期);16794-16803 * |
| Redox-Responsive Magnetic Nanoparticle for Targeted Convection-Enhanced Delivery of O-6-Benzylguanine to Brain Tumors;Stephen ZR，等;ACS NANO;第8卷(第10期);10383-10395 * |
| The design strategy of intelligent biomedical magnesium with controlled release platform;Bi YZ等;MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS;第97卷;254-263 * |
| 响应性纳米药物递送系统构建及肿瘤治疗研究;戴亮亮等;医药生物力学;第34卷;27-28 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112546027B (en) | 2022-08-23 |
| CN115192543A (en) | 2022-10-18 |
| CN112546027A (en) | 2021-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115192543B (en) | Preparation method of lipid-loaded soluble pigment nanoparticles | |
| Manatunga et al. | pH responsive controlled release of anti-cancer hydrophobic drugs from sodium alginate and hydroxyapatite bi-coated iron oxide nanoparticles | |
| CN111265533B (en) | Preparation method of core-shell nano-particles based on lipid membrane and metal organic framework | |
| Xu et al. | Gelatin–mesoporous silica nanoparticles as matrix metalloproteinases-degradable drug delivery systems in vivo | |
| CN114452268B (en) | Preparation method of mesoporous polydopamine anthocyanin-carrying nanoparticle | |
| CN115040496B (en) | Preparation method of hollow mesoporous polydopamine lipid-loaded soluble pigment nanoparticles | |
| Chen et al. | Chitosan-based oral colon-specific delivery systems for polyphenols: recent advances and emerging trends | |
| Braga | Cyclodextrin superstructures for drug delivery | |
| CN114392248B (en) | Preparation method of anthocyanin-carrying nanoparticles | |
| Walia et al. | A bimodal molecular imaging probe based on chitosan encapsulated magneto-fluorescent nanocomposite offers biocompatibility, visualization of specific cancer cells in vitro and lung tissues in vivo | |
| Pourmadadi et al. | Synthesis and characterization of biological macromolecules double emulsion based on carboxymethylcellulose/gelatin hydrogel incorporated with ZIF-8 as metal organic frameworks for sustained anti-cancer drug release | |
| Najafi et al. | Formulation of double nanoemulsions based on pH-sensitive carboxymethyl cellulose/Starch copper doped carbon quantum dots for quercetin controlled release | |
| Kumar et al. | Therapeutic nanomaterials: from a drug delivery perspective | |
| CN109953974B (en) | Preparation method of enzyme-reduction dual-responsiveness hyaluronic acid-polypropylene sulfide copolymer nanocapsule | |
| CN108938593B (en) | Preparation method of lipid-coated solid drug nanoparticles | |
| CN113244234B (en) | Aureoamidol ester-barbaloin-palmitate self-assembled nanoparticles and preparation method and application thereof | |
| Bao et al. | Taste masking of a drug by pH-responsive coordination polymer-coated mesoporous silica nanoparticles | |
| Shashidhara et al. | Pectin nanoforms—a multifaceted polysaccharide and a propitious nanocarrier for medical ailments | |
| CN113476425A (en) | PH-responsive enrofloxacin nanoparticles and preparation method and application thereof | |
| CN110327311A (en) | A kind of preparation method of oleanolic acid slow release nanometer microcapsules | |
| CN116270549B (en) | Colitis-targeted matrine oxide nanoparticle, and preparation method and application thereof | |
| CN114470229B (en) | Preparation and application of carrier-free double-drug self-assembled nanoparticle | |
| CN111888460B (en) | Preparation method of nano drug-loaded micelle with daptomycin-entrapped small-molecule hydrophobic drug | |
| CN110591016B (en) | MAA-co-DAA-co-PEGMA copolymer and preparation method and application thereof | |
| US20240122874A1 (en) | Oral anti-radiation microalgae-nanoparticle compound preparation, and preparation method and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20221018 Assignee: Zibo Hefeng Seed Technology Co.,Ltd. Assignor: HUAIYIN INSTITUTE OF TECHNOLOGY Contract record no.: X2023990000959 Denomination of invention: Preparation method of lipid soluble pigment nanoparticles Granted publication date: 20230630 License type: Common License Record date: 20231205 |
|
| EE01 | Entry into force of recordation of patent licensing contract |