CN115054699B - 一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 - Google Patents
一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 Download PDFInfo
- Publication number
- CN115054699B CN115054699B CN202210545823.7A CN202210545823A CN115054699B CN 115054699 B CN115054699 B CN 115054699B CN 202210545823 A CN202210545823 A CN 202210545823A CN 115054699 B CN115054699 B CN 115054699B
- Authority
- CN
- China
- Prior art keywords
- pei
- plga
- mir
- peg
- solution
- 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
- 108091061970 miR-26a stem-loop Proteins 0.000 title claims abstract description 36
- 239000003937 drug carrier Substances 0.000 title claims abstract description 24
- 210000004185 liver Anatomy 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 230000008685 targeting Effects 0.000 title description 7
- 238000000502 dialysis Methods 0.000 claims abstract description 20
- 238000004108 freeze drying Methods 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 239000008367 deionised water Substances 0.000 claims abstract description 11
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 11
- 239000011258 core-shell material Substances 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 claims abstract 2
- 229920001223 polyethylene glycol Polymers 0.000 claims description 36
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 32
- 239000000243 solution Substances 0.000 claims description 28
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 26
- 229920002674 hyaluronan Polymers 0.000 claims description 23
- 229960003160 hyaluronic acid Drugs 0.000 claims description 23
- 238000003756 stirring Methods 0.000 claims description 15
- 239000002202 Polyethylene glycol Substances 0.000 claims description 14
- 229920001577 copolymer Polymers 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 7
- 230000003213 activating effect Effects 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 5
- 229920002385 Sodium hyaluronate Polymers 0.000 claims description 4
- 239000012190 activator Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 229940010747 sodium hyaluronate Drugs 0.000 claims description 4
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 2
- 239000003054 catalyst Substances 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 238000007710 freezing Methods 0.000 claims description 2
- 230000008014 freezing Effects 0.000 claims description 2
- 208000014018 liver neoplasm Diseases 0.000 abstract description 26
- 201000007270 liver cancer Diseases 0.000 abstract description 25
- 238000006243 chemical reaction Methods 0.000 abstract description 10
- 238000001727 in vivo Methods 0.000 abstract description 10
- 108091070501 miRNA Proteins 0.000 abstract description 7
- 239000002679 microRNA Substances 0.000 abstract description 7
- 238000000338 in vitro Methods 0.000 abstract description 6
- 229920006317 cationic polymer Polymers 0.000 abstract description 4
- 231100000053 low toxicity Toxicity 0.000 abstract description 2
- 108091046553 miR-26 stem-loop Proteins 0.000 abstract description 2
- 108091023821 miR-26-1 stem-loop Proteins 0.000 abstract description 2
- 108091045094 miR-26-2 stem-loop Proteins 0.000 abstract description 2
- 238000001338 self-assembly Methods 0.000 abstract description 2
- 150000001408 amides Chemical class 0.000 abstract 2
- 229920002873 Polyethylenimine Polymers 0.000 description 36
- 210000004027 cell Anatomy 0.000 description 25
- 239000003814 drug Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 150000007523 nucleic acids Chemical group 0.000 description 9
- 229940079593 drug Drugs 0.000 description 7
- 239000002539 nanocarrier Substances 0.000 description 7
- 231100000135 cytotoxicity Toxicity 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 6
- 239000002105 nanoparticle Substances 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000002114 nanocomposite Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 239000012980 RPMI-1640 medium Substances 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 101150108055 CHMP2B gene Proteins 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 2
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 230000004700 cellular uptake Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000012637 gene transfection Methods 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 210000003712 lysosome Anatomy 0.000 description 2
- 230000001868 lysosomic effect Effects 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 102000016736 Cyclin Human genes 0.000 description 1
- 108050006400 Cyclin Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- -1 HA sodium salt Chemical class 0.000 description 1
- 206010019695 Hepatic neoplasm Diseases 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 101150099493 STAT3 gene Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000004900 autophagic degradation Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000035572 chemosensitivity Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 238000001476 gene delivery Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000006676 mitochondrial damage Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 210000000865 mononuclear phagocyte system Anatomy 0.000 description 1
- 230000009437 off-target effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229920001601 polyetherimide Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 150000003511 tertiary amides Chemical class 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- 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
- A61K47/58—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 by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
-
- 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
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- 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
- A61K47/593—Polyesters, e.g. PLGA or polylactide-co-glycolide
-
- 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
- A61K47/51—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
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Nanotechnology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种肝靶向递送miR‑26a类似物的纳米药物载体及其制备方法,采用三次酰胺反应,可将不同分子量的PLGA、PEG、HA接枝到阳离子聚合物支化PEI上,通过透析、过滤、冻干等方法得到用于miRNA体外及体内递送的载体PLGA‑PEI‑PEG‑HA。递送载体在去离子水中水化后,自组装形成球型的核壳结构,表面电荷为40~60mV,能以N/P比为20:1完全有效地包载miR‑26amimics,可用于体外和体内低毒高效地主动递送至肝癌细胞。
Description
技术领域
本发明属于药物载体技术领域,具体涉及到一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法。
背景技术
近年来,在大量临床样本检测中发现miR-26a的表达水平变化与肝炎、肝硬化、肝癌呈显著相关,证实miR-26a可作为临床早期鉴定及肝癌分期的生物标志物。同时,临床数据分析显示,与miR-26a表达水平较低的患者相比,miR-26a表达水平高的患者总生存期和复发时间更长。针对临床肝细胞癌(HCC)细胞显著低表达的miR-26a,许多研究者探索了miR-26a与HCC发生发展的分子机制,证明了miR-26a通过介导肝癌发生发展各通路的关键因子的表达,如细胞周期蛋白家族、IL-6/Stat3、生长因子等,来抑制肝癌细胞的增殖、迁移、血管生成等,并且miR-26可通过抑制细胞自噬,增强化学敏感性,促进肝癌细胞凋亡。鉴于miR-26a对肝癌发展的抑制作用,外源补充miR-26a,使肝癌细胞恢复miR-26a水平,从而抑制癌细胞生长,成为了一种前景良好的肝癌治疗手段。
近年来随着高分子材料领域的不断发展,越来越多的聚合物分子被证明具有良好的基因递送潜力。聚乙烯亚胺(PEI)是基础研究中被广泛用于纳米药物载体构建的聚合物。PEI是一种水溶性高分子聚合物,具有质子海绵效应,即进入细胞后引起溶酶体破裂从而导致线粒体损伤和细胞凋亡,被广泛用于基因转染和药物递送中。该聚合物具有超支化结构,并带有许多伯胺基团,这些伯胺基团在生理条件下被质子化。PEI的高正电荷特性可使该聚合物与带负电的细胞膜发生静电相互作用,从而促进细胞摄取。
聚乳酸-羟基乙酸共聚物(PLGA)是美国FDA批准的药用辅料,在体内可经过三羧酸循环最后降解为水和二氧化碳,具有良好的生物相容性和生物降解性,此外,PLGA还具有良好的成囊成膜的性能,广泛用于制药、医用工程材料等。研究表明,经PLGA修饰后的PEI(PLGA-PEI)纳米粒的基因转染效果优于PEI纳米粒,可以显著降低PEI的细胞毒性。
聚乙二醇(PEG)是最常用亲水性修饰材料,也是药物载体系统想载体材料之一,其免疫原性及抗原性低,能有效屏蔽阳离子聚合物纳米粒的部分正电荷,降低聚阳离子的细胞毒性,同时降低网状内皮系统的识别和吞噬。PEG的亲水性有利于延长载体在体内的循环时间,增强载体在血液中的稳定性,并有利于通过“增强渗透与滞留”(EPR)效应增加肿瘤组织的蓄积量。对于载生物药物的纳米粒,PEG可以屏蔽生物药物的免疫原性,减少外界条件(酸、碱、酶、水和热等)对生物药物活性的影响,延长生物药物在体内的半衰期。
透明质酸(HA)是一种酸性粘多糖,能识别肿瘤细胞表面高表达的CD44受体,与之结合使药物入胞,成为介导药物细胞膜内转运的非常有潜力的物质。已有研究表明,HA修饰的PEI阳离子纳米粒能靶向肿瘤细胞,显著提高肿瘤药物的细胞摄取,提高药物疗效。
不断深入的研究发现,miRNA药物的体内递送面临许多挑战。例如,要实现组织特异性递送、靶细胞的有效摄取;要避免核酸酶的降解、脱靶效应、免疫系统激活;要降低递送材料的毒副作用等。
发明内容
本发明的目的是提供一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法,可以利用其透明质酸靶向于肝脏肿瘤部位,将基因药物miR-26a类似物(miR-26amimics)递送至肝癌细胞,实现靶点可控定位释药,从而恢复肝癌细胞miR-26a水平,达到抑制肝癌发展的治疗作用。
为达上述目的,本发明提供了一种肝靶向递送miR-26a类似物的纳米药物载体的制备方法,包括以下步骤:
(1)将b-PEI溶液滴加于活化后的聚乳酸-羟基乙酸共聚物溶液中,恒温搅拌后经透析、过滤和冷冻干燥,制得PLGA-PEI;
(2)将步骤(1)制得的PLGA-PEI与聚乙二醇共溶于二甲基亚砜中,加入活化剂,恒温搅拌后经透析、过滤和冷冻干燥,制得PLGA-PEI-PEG;
(3)将透明质酸溶于二甲基亚砜中,加入活化剂,室温搅拌后制得活化的透明质酸溶液;
(4)将步骤(2)制得的PLGA-PEI-PEG与步骤(3)制得的透明质酸溶液混合后,恒温搅拌后经透析、冷冻干燥后制得,命名为PLGA-PEI-PEG-HA。
进一步地,步骤(1)中的活化后的聚乳酸-羟基乙酸共聚物溶液通过以下方法制备得到:将聚乳酸-羟基乙酸共聚物溶于二甲基亚砜中,加入活化剂后室温搅拌3-5h,制得;聚乳酸-羟基乙酸共聚物与活化剂的质量比为3.5-4.0:0.85。
进一步地,b-PEI(枝状聚乙烯亚胺Branched polyethylenimine,b-PEI)溶液通过以下方法制备得到:将b-PEI溶于二甲基亚砜溶液中,于室温下搅拌3-5h,制得;b-PEI溶解后的浓度为0.08-0.1g/mL。
进一步地,步骤(1)、步骤(2)和步骤(4)中恒温的温度为30-40℃,搅拌的时间为20-25h,透析的时间为45-50h,冷冻干燥的温度为-80℃,冷冻干燥的时间为2h以上。
进一步地,活化剂为1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐和N-羟基丁二酰亚胺按照质量比8-250:5-150的混合物。
进一步地,步骤(2)中的PLGA-PEI与聚乙二醇的质量比为3.3:0.15-0.17,所述聚乙二醇溶于二甲基亚砜后的浓度为2.0*10-3-2.1*10-3g/mL。
进一步地,步骤(3)中的透明质酸通过以下方法制备得到:将透明质酸钠盐溶解于去离子水中,以pH为3.5的盐酸溶液为透析液,透析10-14h后,取透析袋内混合液于-80℃冷冻2h以上,制得;
溶解于去离子水中的透明质酸钠盐浓度为0.01-0.03g/mL。
进一步地,步骤(4)中PLGA-PEI-PEG与透明质酸溶液中透明质酸的质量比为1.5-2.0:0.07。
进一步地,聚乳酸-羟基乙酸共聚物溶液的分子量为6-36kDa。
本发明还提供了一种肝靶向递送miR-26a类似物的纳米药物载体的制备方法制备得到的PLGA-PEI-PEG-HA。
采用上述方案的有益效果是:该纳米药物载体可以递送miR-26a类似物,并且具有肝靶向作用。
综上所述,本发明具有以下优点:
1、本发明所用的纳米复合物形成的原料生物安全性均良好,且具有生物可降解性,已被广泛应用于生物医药领域。
2、通过本发明制备的肝靶向递送miR-26a类似物的纳米药物载体为具有包载肝癌抑制功能的miRNA的纳米复合物,制备过程简便温和,制备原料廉价易得,载体粒径范围适宜。
3、本发明制备的纳米药物载体在小鼠肝癌模型上应用时,可采用尾静脉注射的方式,给药后纳米复合物通过肝癌的高渗透性和滞留效应被动蓄积到肿瘤部位。载体表面的HA可主动靶向肝癌细胞,使载体被靶细胞摄取,并且基于合成的递送载体的高入胞和内涵体/溶酶体逃逸能力可实现目标miRNA(miR-26a mimics)的高效递送,在miRNA的肝癌治疗方面有良好的应用前景。
4、本发明采用三次酰胺反应,可将不同分子量的PLGA、PEG、HA接枝到阳离子聚合物支化PEI上,通过透析、过滤、冻干等方法得到用于miRNA体外及体内递送的载体PLGA-PEI-PEG-HA。
递送载体在去离子水中水化后,自组装形成球型的核壳结构,表面电荷为40~60mV,能以N/P比为20:1完全有效地包载miR-26a mimics,可用于体外和体内低毒高效地主动递送至肝癌细胞。
附图说明
图1为PLGA-PEI、PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的透射电镜图;
其中A为PLGA-PEI;B为PLGA-PEI-PEG,C为PLGA-PEI-PEG-HA;
图2为PLGA-PEI的核磁共振氢谱;
图3为PLGA-PEI-PEG的核磁共振氢谱;
图4为PLGA-PEI-PEG-HA的核磁共振氢谱;
图5为PLGA-PEI、PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的傅里叶变换红外光谱图;
图6为PLGA-PEI、PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的粒径和电位示意图;
图7为PLGA-PEI、PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的PEI结枝率;
其中由上至下依次为结合HA(%)、结合PEG(%)、结合PLGA(%)和游离PEI(%);
图8为PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的琼脂糖凝胶电泳图像:
图9和图10为PLGA-PEI-PEG、PLGA-PEI-PEG-HA纳米载体的细胞毒性的实验结果;
图11为PLGA-PEI-PEG、PLGA-PEI-PEG-HA载体的琼脂糖凝胶电泳图像;
图12为PLGA-PEI-PEG、PLGA-PEI-PEG-HA载体的细胞毒性实验结果;
图13为实施例4中PLGA24kDa-PEI-PEG、PLGA24kDa-PEI-PEG-HA载体包载si-CHMP2B转染SNU449细胞后(载体包载si-DHX36转染Huh7细胞后),基因CHMP2B(DHX36)的蛋白表达情况。
具体实施方式
以下结合实施例对本发明的原理和特征进行描述,所举实例只用于解释本发明,并非用于限定本发明的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
实施例1
本实施例提供了PLGA-PEI的制备方法,包括以下步骤:
(1)将0.3840g PLGA(Mw=24kDa)溶于5mL DMSO,加入0.0530gEDC·HCl和0.0320gNHS,于室温下搅拌4h,活化PLGA的羧基基团,制得PLGA反应液。
(2)将0.400g b-PEI溶于另一5mL的DMSO溶液中,于室温下搅拌4h后于搅拌下缓慢滴入PLGA反应液中,恒温35℃搅拌反应24h。
(3)将步骤(2)的反应液转至去离子水中透析(MWCO=25kDa)48h,期间多次更换透析液。
(4)透析结束后,将透析袋内混合液过滤(0.45μm),收集滤液置于-80℃冷冻2h以上,经冷冻干燥获得PLGA-PEI。
实施例2
本实施例提供了PLGA-PEI-PEG的制备方法,包括以下步骤:
(1)将0.3300g PLGA-PEI、0.0165g PEG(2kDa)溶于8mL DMSO,0.2190g EDC·HCl和0.1320g NHS,于恒温35℃搅拌反应24h。
(2)反应结束后,将反应液转至去离子水中透析(MWCO=2kDa)48h,期间多次更换透析液。
(3)透析结束后,将透析袋内混合液过滤(0.45μm),收集滤液置于-80℃冷冻2h以上,再经冷冻干燥获得PLGA-PEI-PEG。
实施例3
本实施例提供了PLGA-PEI-PEG-HA的制备方法,包括以下步骤:
(1)将0.100g HA钠盐溶解在5mL去离子水中,并使用MWCO=2kDa透析袋对稀HCl溶液(pH=3.5)透析12h,期间多次更换透析液。
(2)透析结束后,将透析袋内混合液置于-80℃冷冻2h以上,经冷冻干燥获得HA。
(3)将冻干所得的HA取0.0070g溶于5mL DMSO中,加入0.0093g EDC·HC、0.0056gNHS,室温搅拌2h,制得活化的HA。
(4)将冻干所得的PLGA-PEI-PEG取0.1785g加入活化的HA中,于恒温35℃搅拌反应24h。
(5)反应结束后,将反应液转至去离子水中透析(MWCO=2kDa),期间多次更换透析液。
(6)透析结束后,将透析袋内混合液置于-80℃冷冻2h以上,经冷冻干燥获得PLGA-PEI-PEG-HA。
本实施例中的PLGA的分子量可按反应比例做出相应调整。例如,使用6kDa、12kDa、24kDa、36kDa的PLGA均可通过上述制备步骤获得PLGA-PEI-PEG-HA载体(如图11)。
试验例1
PLGA-PEI-PEG-HA包载miR-26a mimics对肝癌细胞的体外和体内递送检测试验
S1:分别对实施例1~3制备得到的纳米载体进行透射电子显微镜检测(如图1),傅里叶变换红外光谱分析(如图5),核磁共振氢谱分析(如图2-4),粒径与电位测定(如图6)。
S2:分别对实施例1~3制备得到的纳米药物载体进行PEI结枝率测定(如图7)、miRNA包载效率测定(如图8)、细胞毒性测试(如图9-10)、转染效率测定(如图13)测试,筛选得到转染效率最高且细胞毒性最低的递送载体,即可用于体外递送miR-26a mimics以达到抑制肝癌细胞增殖和迁移的作用;用于体内递送mIR-26a mimics以发挥肝癌的治疗作用。
从图1-7可以看出,PLGA、PEG、HA三部分成功接枝到阳离子聚合物PEI上,合成的递送载体具有明显且规整的球形的核壳结构,粒径约为150~250nm,表面Zeta电位约为40~60mV,有利于核酸片段的包载。
从图8中可以看出,PLGA-PEI-PEG、PLGA-PEI-PEG-HA载体与包载的核酸片段的N/P比在10:1及以上时,核酸片段可被完全有效地包载。
从图11-12可以看出,当PLGA分子量为24kDa时,PLGA-PEI-PEG-HA载体以N/P=20:1包载核酸片段,载体的毒性最低。其中PPP24载体即为PLGA(24kDa)-PEI-PEG,HA-PPP载体即为PLGA(24kDa)-PEI-PEG-HA载体。
试验例2
PLGA-PEI-PEG-HA载体包载核酸片段si-CHMP2B对肝癌细胞SNU499、包载核酸片段si-DHX36对肝癌细胞Huh7的递送效果的验证
S1:按照实施例1~3的制备方法合成PLGA(24kDa)-PEI-PEG、PLGA(24kDa)-PEI-PEG-HA载体,分别将其分散在去离子水中,形成一定浓度(1mg/mL或10mg/mL)的分散液。将合成载体以不同的N/P比与一定量的核酸片段si-CHMP2B(或si-DHX36)混合后常温静置孵育30min,使合成载体通过静电吸附作用包载核酸片段。
S2:将处于对数生长期且生长状态良好的肝癌细胞SNU449(Huh7),加入胰蛋白酶消化,RPMI 1640(DMEM)完全培养基终止消化后,将细胞悬液离心,弃去上清,用RPMI 1640(DMEM)完全培养基重悬后接种于24孔板中。
S3:待细胞贴壁后,弃去培养液,分别加入用RPMI 1640(DMEM)培养基稀释纳米复合物si-CHMP2B@PLGA-PEI-PEG、si-DHX36@PLGA-PEI-PEG-HA,孵育36h后,收取细胞蛋白,通过检测基因CHMP2B(或DHX36)的表达来反映出合成载体对肝癌细胞SNU449(或Huh7)的递送效果。其中,N/P比为PEI游离氨基的摩尔数与核酸片段磷酸基团的摩尔数之比。
实验结果:如图13所示,在N/P比为20:1~30:1时,PLGA-PEI-PEG、PLGA-PEI-PEG-HA载体均表现出对肝癌细胞SNU449(或Huh7)良好的递送效果。
虽然对本发明的具体实施方式进行了详细地描述,但不应理解为对本专利的保护范围的限定。在权利要求书所描述的范围内,本领域技术人员不经创造性劳动即可作出的各种修改和变形仍属本专利的保护范围。
Claims (6)
1.一种肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述肝靶向递送miR-26a类似物的纳米药物载体为PLGA、PEG和HA均接枝到b-PEI并成核壳结构;
所述肝靶向递送miR-26a类似物的纳米药物载体的制备包括以下步骤:
(1)将b-PEI溶液滴加于活化后的聚乳酸-羟基乙酸共聚物溶液中,恒温搅拌后经透析、过滤和冷冻干燥,制得PLGA-PEI;
(2)将步骤(1)制得的PLGA-PEI与聚乙二醇共溶于二甲基亚砜中,加入活化剂,恒温搅拌后经透析、过滤和冷冻干燥,制得PLGA-PEI-PEG;
(3)将透明质酸溶于二甲基亚砜中,加入活化剂,室温搅拌后制得活化的透明质酸溶液;
(4)将步骤(2)制得的PLGA-PEI-PEG与步骤(3)制得的透明质酸溶液混合后,恒温搅拌后经透析、冷冻干燥后制得;
所述步骤(1)中的活化后的聚乳酸-羟基乙酸共聚物溶液通过以下方法制备得到:将聚乳酸-羟基乙酸共聚物溶于二甲基亚砜中,加入活化剂后室温搅拌3-5h,制得;所述聚乳酸-羟基乙酸共聚物与活化剂的质量比为3.5-4.0:0.85;
所述步骤(2)中的PLGA-PEI与聚乙二醇的质量比为3.3:0.15-0.17,所述聚乙二醇溶于所述二甲基亚砜后的浓度为2.0*10-3-2.1*10-3g/mL;
所述步骤(4)中PLGA-PEI-PEG与透明质酸溶液中透明质酸的质量比为1.5-2.0:0.07;
所述miR-26a类似物为si-DHX36。
2.如权利要求1所述的肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述b-PEI溶液通过以下方法制备得到:将b-PEI溶于二甲基亚砜溶液中,于室温下搅拌3-5h,制得;所述b-PEI溶解后的浓度为0.08-0.1g/mL。
3.如权利要求1所述的肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述步骤(1)、步骤(2)和步骤(4)中恒温的温度为30-40℃,搅拌的时间为20-25h,透析的时间为45-50h,冷冻干燥的温度为-80℃,冷冻干燥的时间为2h以上。
4.如权利要求1所述的肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述活化剂为1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐和N-羟基丁二酰亚胺按照质量比8-250:5-150的混合物。
5.如权利要求1所述的肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述步骤(3)中的透明质酸通过以下方法制备得到:将透明质酸钠盐溶解于去离子水中,以pH为3.5的盐酸溶液为透析液,透析10-14h后,取透析袋内混合液于-80℃冷冻2h以上,制得;
所述溶解于去离子水中的透明质酸钠盐浓度为0.01-0.03g/mL。
6.如权利要求1所述的肝靶向递送miR-26a类似物的纳米药物载体,其特征在于,所述聚乳酸-羟基乙酸共聚物溶液的分子量为6-36kDa。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210545823.7A CN115054699B (zh) | 2022-05-18 | 2022-05-18 | 一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210545823.7A CN115054699B (zh) | 2022-05-18 | 2022-05-18 | 一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115054699A CN115054699A (zh) | 2022-09-16 |
CN115054699B true CN115054699B (zh) | 2024-01-26 |
Family
ID=83198157
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210545823.7A Active CN115054699B (zh) | 2022-05-18 | 2022-05-18 | 一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115054699B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116425973B (zh) * | 2023-04-03 | 2024-01-26 | 深圳市华元生物技术股份有限公司 | 一种pH响应型的肝靶向的药物递送载体及其制备方法和应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105169400A (zh) * | 2015-08-10 | 2015-12-23 | 东华大学 | 一种透明质酸靶向的多功能支化聚乙烯亚胺药物载体的制备方法 |
CN105646887A (zh) * | 2016-01-05 | 2016-06-08 | 湖北大学 | 一种两亲性高分子聚合物及其制备方法、应用 |
CN105873613A (zh) * | 2013-08-13 | 2016-08-17 | 贝勒医学院 | 用于核酸和药物递送的新型plga-修饰的聚乙烯亚胺自组装纳米技术 |
CN107715121A (zh) * | 2017-09-19 | 2018-02-23 | 暨南大学 | 一种磁共振成像纳米药物载体、纳米载药系统及其制备方法 |
-
2022
- 2022-05-18 CN CN202210545823.7A patent/CN115054699B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105873613A (zh) * | 2013-08-13 | 2016-08-17 | 贝勒医学院 | 用于核酸和药物递送的新型plga-修饰的聚乙烯亚胺自组装纳米技术 |
CN105169400A (zh) * | 2015-08-10 | 2015-12-23 | 东华大学 | 一种透明质酸靶向的多功能支化聚乙烯亚胺药物载体的制备方法 |
CN105646887A (zh) * | 2016-01-05 | 2016-06-08 | 湖北大学 | 一种两亲性高分子聚合物及其制备方法、应用 |
CN107715121A (zh) * | 2017-09-19 | 2018-02-23 | 暨南大学 | 一种磁共振成像纳米药物载体、纳米载药系统及其制备方法 |
Also Published As
Publication number | Publication date |
---|---|
CN115054699A (zh) | 2022-09-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Park et al. | Advances in the synthesis and application of nanoparticles for drug delivery | |
Jin et al. | Current progress in gene delivery technology based on chemical methods and nano-carriers | |
CN105727307B (zh) | 一种硫辛酸修饰的纳米多肽载体及其制备方法和应用 | |
CN110801431B (zh) | 一种核-壳型智能纳米递送系统的构建及应用 | |
Zheng et al. | Biodegradable and redox-responsive chitosan/poly (L-aspartic acid) submicron capsules for transmucosal delivery of proteins and peptides | |
Li et al. | Advances and potential applications of chitosan nanoparticles as a delivery carrier for the mucosal immunity of vaccine | |
Laroui et al. | Gastrointestinal delivery of anti-inflammatory nanoparticles | |
CN109498548B (zh) | 一种蛋白质与光敏剂共传递pH响应性聚氨基酸纳米凝胶及其制备方法 | |
CN104262638A (zh) | 透明质酸-胱胺-聚乳酸-羟基乙酸接枝聚合物及其制备方法 | |
CN104586816A (zh) | 还原触发型多肽修饰透明质酸偶联物载体及其制备 | |
Huang et al. | Design and application of dextran carrier | |
CN115054699B (zh) | 一种肝靶向递送miR-26a类似物的纳米药物载体及其制备方法 | |
CN112168975A (zh) | 一种抗肿瘤靶向药物缓释载体、制剂及其制备方法 | |
Wang et al. | Applications and functions of γ-poly-glutamic acid and its derivatives in medicine | |
CN109662956B (zh) | 一种齐墩果酸接枝的壳聚糖载药纳米颗粒的应用 | |
Song et al. | Preparation and evaluation of insulin-loaded nanoparticles based on hydroxypropyl-β-cyclodextrin modified carboxymethyl chitosan for oral delivery | |
CN115337272B (zh) | 一种天然多糖基化学-物理双交联水凝胶微粒及其制备与应用 | |
EP2907876B1 (en) | Reduction stimuli-responsive gene vector system and preparation and use thereof | |
Blanco-Fernandez et al. | Fabrication of magnetic and fluorescent chitin and dibutyrylchitin sub-micron particles by oil-in-water emulsification | |
CN113908137B (zh) | 一种注射用硬核软膜型纳米缓释递药系统的制备方法 | |
CN106421812B (zh) | 一种自组装四氧化三铁纳米颗粒的制法和用途 | |
WO2016101405A1 (zh) | 一种安全高效的蛋白质转染进入细胞的载体 | |
CN112843251A (zh) | 细胞穿膜肽修饰的药物载体及其制备方法和应用 | |
CN115260286A (zh) | Dmp-f11多肽偶联物及其制备方法和用途 | |
CN115137846B (zh) | 基因药物或基因疫苗纳米递送系统和应用 |
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 |