CN111632038A - A kind of platelet drug-loaded micro-nano motor and its preparation method and application - Google Patents
A kind of platelet drug-loaded micro-nano motor and its preparation method and application Download PDFInfo
- Publication number
- CN111632038A CN111632038A CN202010509926.9A CN202010509926A CN111632038A CN 111632038 A CN111632038 A CN 111632038A CN 202010509926 A CN202010509926 A CN 202010509926A CN 111632038 A CN111632038 A CN 111632038A
- Authority
- CN
- China
- Prior art keywords
- platelet
- drug
- micro
- nano motor
- preparation
- 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.)
- Granted
Links
- 239000003814 drug Substances 0.000 title claims abstract description 73
- 229940079593 drug Drugs 0.000 title claims abstract description 71
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 229920001690 polydopamine Polymers 0.000 claims abstract description 44
- 239000011248 coating agent Substances 0.000 claims abstract description 32
- 238000000576 coating method Methods 0.000 claims abstract description 32
- 150000001491 aromatic compounds Chemical class 0.000 claims abstract description 7
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 7
- -1 amine compound Chemical class 0.000 claims abstract description 6
- 239000000725 suspension Substances 0.000 claims description 46
- 239000000872 buffer Substances 0.000 claims description 34
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 27
- 229960004679 doxorubicin Drugs 0.000 claims description 13
- 238000005119 centrifugation Methods 0.000 claims description 10
- 239000000203 mixture Substances 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 9
- 229960001149 dopamine hydrochloride Drugs 0.000 claims description 9
- 206010028980 Neoplasm Diseases 0.000 claims description 8
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 7
- 229930192392 Mitomycin Natural products 0.000 claims description 6
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 claims description 6
- 201000011510 cancer Diseases 0.000 claims description 6
- 229960004857 mitomycin Drugs 0.000 claims description 6
- 229960001420 nimustine Drugs 0.000 claims description 6
- VFEDRRNHLBGPNN-UHFFFAOYSA-N nimustine Chemical compound CC1=NC=C(CNC(=O)N(CCCl)N=O)C(N)=N1 VFEDRRNHLBGPNN-UHFFFAOYSA-N 0.000 claims description 6
- 150000001412 amines Chemical class 0.000 claims description 5
- 210000004369 blood Anatomy 0.000 claims description 5
- 239000008280 blood Substances 0.000 claims description 5
- 206010027476 Metastases Diseases 0.000 claims description 4
- 230000009401 metastasis Effects 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 210000003743 erythrocyte Anatomy 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 230000033001 locomotion Effects 0.000 abstract description 10
- 239000011859 microparticle Substances 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 7
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 5
- 238000001089 thermophoresis Methods 0.000 abstract description 5
- 206010061218 Inflammation Diseases 0.000 abstract description 4
- 230000004054 inflammatory process Effects 0.000 abstract description 4
- 229960003638 dopamine Drugs 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 238000011065 in-situ storage Methods 0.000 abstract description 2
- 230000020411 cell activation Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 25
- 239000000243 solution Substances 0.000 description 8
- 238000012377 drug delivery Methods 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 description 6
- 229960002918 doxorubicin hydrochloride Drugs 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 230000035515 penetration Effects 0.000 description 5
- 206010052428 Wound Diseases 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 238000004163 cytometry Methods 0.000 description 4
- 230000008827 biological function Effects 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 230000035605 chemotaxis Effects 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 239000002262 Schiff base Substances 0.000 description 2
- 150000004753 Schiff bases Chemical class 0.000 description 2
- 208000002847 Surgical Wound Diseases 0.000 description 2
- 208000007536 Thrombosis Diseases 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000004700 cellular uptake Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000003399 chemotactic effect Effects 0.000 description 2
- 229940044683 chemotherapy drug Drugs 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- 210000004623 platelet-rich plasma Anatomy 0.000 description 2
- 238000002271 resection Methods 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000010118 platelet activation Effects 0.000 description 1
- 230000000379 polymerizing effect Effects 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 210000001243 pseudopodia Anatomy 0.000 description 1
- 239000002520 smart material Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000002525 ultrasonication 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
- A61K9/5192—Processes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
-
- 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
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0042—Photocleavage of drugs in vivo, e.g. cleavage of photolabile linkers in vivo by UV radiation for releasing the pharmacologically-active agent from the administered agent; photothrombosis or photoocclusion
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
- A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
- A61K47/6935—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
-
- 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
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
-
- 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
- A61K9/5176—Compounds of unknown constitution, e.g. material from plants or animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nanotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Optics & Photonics (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Zoology (AREA)
- Dermatology (AREA)
- Immunology (AREA)
- Oncology (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种血小板载药微纳米马达及其制备方法和应用，该微纳米马达包括血小板外侧包裹的聚多巴胺涂层以及与聚多巴胺结合的含胺类化合物或芳香族化合物药物分子。本发明利用多巴胺在弱碱性条件下易于自聚合的特点，在血小板表面原位聚合形成聚多巴胺涂层，再利用聚多巴胺与药物分子结合装载药物，形成血小板载药微纳米马达。本发明利用血小板本身的不对称性，在近红外光的照射下，具有光热效应的聚多巴胺涂层可形成不均匀的热泳现象，推动血小板载药微纳米马达的运动。本发明制备方法原料简单、反应条件温和、操作简便，制得的材料具有良好的生物相容性可以自主趋向炎症或伤口部位，并通过血小板细胞活化产生血小板衍生微粒，从而释放药物。
The invention discloses a platelet drug-carrying micro-nano motor and a preparation method and application thereof. The micro-nano motor comprises a polydopamine coating wrapped on the outside of the platelet and a drug molecule containing an amine compound or an aromatic compound combined with the polydopamine. The invention utilizes the characteristics of easy self-polymerization of dopamine under weak alkaline conditions, in-situ polymerization of the platelet surface forms a polydopamine coating, and then the polydopamine is combined with drug molecules to load the drug to form the platelet drug-loaded micro-nano motor. The invention utilizes the asymmetry of the platelet itself, and under the irradiation of near-infrared light, the polydopamine coating with photothermal effect can form an uneven thermophoresis phenomenon and promote the movement of the platelet drug-loaded micro-nano motor. The preparation method of the invention has simple raw materials, mild reaction conditions and simple operation, and the prepared material has good biocompatibility and can autonomously tend to inflammation or wound sites, and generates platelet-derived microparticles through platelet cell activation, thereby releasing drugs.
Description
技术领域technical field
本发明属于生物医学材料，具体涉及一种血小板载药微纳米马达及其制备方法和应用。The invention belongs to biomedical materials, in particular to a platelet drug-carrying micro-nano motor and a preparation method and application thereof.
背景技术Background technique
近年来，多功能的人工合成微纳米马达在催化降解、生物传感、毒素吸附和药物运输等领域表现出良好的应用潜能。然而，为了实现微纳米马达在生物体内的实际应用，需要设计开发完全生物兼容的智能材料，使合成的微纳米马达能够在复杂生物环境中发挥作用，同时不产生有害的影响。In recent years, multifunctional synthetic micro-nanomotors have shown promising potential in the fields of catalytic degradation, biosensing, toxin adsorption, and drug delivery. However, in order to realize the practical application of micro-nanomotors in living organisms, it is necessary to design and develop fully biocompatible smart materials that enable the synthesized micro-nanomotors to function in complex biological environments without harmful effects.
部分天然细胞和微生物具有固有的趋化特性和生物功能。目前已有研究将这些生物组件与纳米材料相结合制造细胞基微纳米马达，用于生物环境的操作和执行各种生物功能。细胞基微纳米马达能够充分发挥生物组件固有的趋化特性，促进微纳米马达在复杂的生物环境中有效运动；并模仿天然细胞的生物功能，在特定位置发挥作用。细胞基微米马达由于不需要额外的燃料或动力装置，在生物医学领域具有广阔的应用前景。Some natural cells and microorganisms have inherent chemotactic properties and biological functions. There have been studies combining these biological components with nanomaterials to fabricate cell-based micro-nanomotors for the manipulation of biological environments and to perform various biological functions. Cell-based micro-nanomotors can give full play to the inherent chemotactic properties of biological components, promote the efficient movement of micro-nanomotors in complex biological environments; and imitate the biological functions of natural cells to function at specific locations. Cell-based micromotors have broad application prospects in the biomedical field because they do not require additional fuel or power devices.
尽管这类细胞基微米马达具有独特的优点，但它们在体内的应用潜力仍面临重大挑战。其中包括对运动速度与方向的控制以及微米级马达的尺寸限制。例如：巨噬细胞马达尺寸达几十微米，很难将药物有效递送至目标细胞内部，极大地限制其作为药物运输载体的功效。Despite the unique advantages of such cell-based micromotors, their potential for in vivo applications still faces significant challenges. These include control over the speed and direction of motion and size constraints for micron-scale motors. For example, the size of the macrophage motor is tens of microns, which makes it difficult to effectively deliver drugs to the interior of target cells, which greatly limits its efficacy as a drug delivery carrier.
发明内容SUMMARY OF THE INVENTION
发明目的：针对现有技术存在的问题，尤其是现有大多数微纳米马达生物相容性欠佳的问题，本发明提供了一种血小板载药微纳米马达及其制备方法。本发明提供的血小板载药微纳米马达以天然细胞血小板为基材，采用温和的表面改性技术在其表面包裹聚多巴胺涂层，再利用聚多巴胺的邻苯二醌基团与含胺类药物的氨基发生席夫碱反应，或利用聚多巴胺和含氨基或者芳香族药物之间的π-π堆积负载药物。该方法制备工艺简单，材料生物相容性良好，制得的血小板载药微纳米马达可以利用血小板固有的趋化性自主趋向炎症、血栓、手术伤口等病理环境的组织和细胞，通过微纳米马达的运动可以增强细胞摄取和组织渗透，从而提高药物递送效率，在生物医学领域具有广阔的应用前景。Purpose of the invention: Aiming at the problems existing in the prior art, especially the problem of poor biocompatibility of most existing micro-nano motors, the present invention provides a platelet drug-loaded micro-nano motor and a preparation method thereof. The platelet drug-loading micro-nano motor provided by the present invention takes natural cell platelets as the base material, uses a mild surface modification technology to coat the polydopamine coating on the surface, and then utilizes the o-phthaloquinone group of polydopamine and amine-containing drugs. The amino group undergoes a Schiff base reaction, or the π-π stacking between polydopamine and amino-containing or aromatic drugs is used to load drugs. The method has the advantages of simple preparation process and good biocompatibility of materials. The obtained platelet drug-loaded micro-nano motor can use the inherent chemotaxis of platelets to autonomously tend to tissues and cells in pathological environments such as inflammation, thrombus, and surgical wound. The movement of ions can enhance cellular uptake and tissue penetration, thereby improving the efficiency of drug delivery, which has broad application prospects in the biomedical field.
本发明提供一种具有刺激响应性形貌变化且生物相容性良好的血小板微纳米马达，以天然细胞血小板为基材，采用温和的表面改性技术在其表面部分包裹聚多巴胺涂层，一方面可以利用血小板天然的趋化性，自主趋向到手术伤口或炎性环境；另一方面还可以利用聚多巴胺的光热效应，在近红外激光的照射下产生不均匀的热泳现象，驱动血小板及其衍生微粒载药微纳米马达的运动。The invention provides a platelet micro-nano motor with stimuli-responsive morphology change and good biocompatibility. The natural cell platelet is used as the base material, and a polydopamine coating is partially wrapped on its surface by using a mild surface modification technology. On the one hand, the natural chemotaxis of platelets can be used to automatically tend to surgical wounds or inflammatory environments; on the other hand, the photothermal effect of polydopamine can be used to generate uneven thermophoresis under the irradiation of near-infrared lasers, which drives platelets and The motion of its derivative microparticle-loaded micro-nanomotors.
技术方案：为了实现上述目的，如本发明所述一种血小板载药微纳米马达，包括血小板外侧包裹的聚多巴胺涂层以及物理或化学方式与聚多巴胺结合的含胺类化合物或芳香族化合物药物分子，在近红外照射下能够运动，成为一种载药微纳米马达。作为优选，所述聚多巴胺涂层的厚度为0.1-1μm。Technical solution: In order to achieve the above purpose, a platelet drug-loaded micro-nano motor according to the present invention includes a polydopamine coating wrapped on the outside of the platelets and a drug containing amine compounds or aromatic compounds that is physically or chemically combined with polydopamine. Molecules, which can move under near-infrared irradiation, become a drug-loaded micro-nanomotor. Preferably, the thickness of the polydopamine coating is 0.1-1 μm.
本发明所述的血小板载药微纳米马达的制备方法，其特征在于，包括以下步骤：The preparation method of the platelet drug-loaded micro-nano motor according to the present invention is characterized in that, comprising the following steps:
(1)将提取的血小板(PLTs)，分散于PBS缓冲液，得到血小板悬浊液，以防止不必要的血小板活化，室温保存备用；(1) Disperse the extracted platelets (PLTs) in PBS buffer to obtain a platelet suspension to prevent unnecessary platelet activation, and store at room temperature for later use;
(2)将盐酸多巴胺溶解于Tris-HCl缓冲液(pH＝8.5)，并与上述血小板悬浊液按一定体积比混合后，在光照下聚合，得到含有聚多巴胺涂层的血小板微纳米马达悬浊液；离心洗涤后分散于PBS缓冲液保存备用；(2) Dissolving dopamine hydrochloride in Tris-HCl buffer (pH=8.5), mixing with the above-mentioned platelet suspension according to a certain volume ratio, and polymerizing under light to obtain a platelet micro-nano motor suspension containing polydopamine coating Turbid liquid; after centrifugal washing, it is dispersed in PBS buffer and stored for later use;
(3)将含胺类化合物或芳香类化合物药物溶解于PBS缓冲液，与上述含有聚多巴胺涂层的血小板微纳米马达悬浊液在室温下混合振荡，得到负载药物的血小板载药微纳米马达悬浊液；离心洗涤后分散于PBS缓冲液保存备用。(3) Dissolve the amine compound or aromatic compound drug in PBS buffer, mix and shake with the above-mentioned platelet micro-nano motor suspension containing polydopamine coating at room temperature to obtain a drug-loaded platelet drug-loaded micro-nano motor suspension; centrifuged and washed, dispersed in PBS buffer and stored for later use.
其中，所述聚多巴胺涂层的厚度为0.1-1μm。Wherein, the thickness of the polydopamine coating is 0.1-1 μm.
作为优选，所述步骤(1)提取血小板为使用梯度离心法从新鲜全血中分离提取血小板，梯度离心法先用1000-1500rpm离心15-20分钟去除红细胞，再使用2500-3000rpm离心10-15分钟富集血小板。Preferably, the step (1) to extract platelets is to use gradient centrifugation to separate and extract platelets from fresh whole blood. The gradient centrifugation method is firstly centrifuged at 1000-1500rpm for 15-20 minutes to remove red blood cells, and then centrifuged at 2500-3000rpm for 10-15 minutes. Platelet enrichment in minutes.
作为优选，步骤(2)所述盐酸多巴胺的Tris-HCl溶液浓度为0.1-2mg/mL，与血小板悬浊液的体积比为1:1-1:10，血小板悬浊液中血小板密度为1×107-8个/mL，聚合反应时间为2-6h。Preferably, the concentration of the Tris-HCl solution of dopamine hydrochloride in step (2) is 0.1-2 mg/mL, and the volume ratio to the platelet suspension is 1:1-1:10, and the platelet density in the platelet suspension is 1 ×10 7-8 pieces/mL, and the polymerization reaction time is 2-6h.
其中，步骤(4)所述药物为含胺类化合物或芳香类化合物，包括如阿霉素、尼莫司汀、丝裂霉素等。Wherein, the medicine described in step (4) is an amine-containing compound or an aromatic compound, including, for example, doxorubicin, nimustine, mitomycin and the like.
作为优选，步骤(3)所述药物浓度为0.1-2mg/mL，与含有聚多巴胺涂层的血小板微纳米马达悬浊液的体积比为1:1-5:1，其中血小板密度为1×107-8个/mL，与含有聚多巴胺涂层的血小板微纳米马达悬浊液在室温下混合2-12小时。Preferably, the concentration of the drug in step (3) is 0.1-2 mg/mL, and the volume ratio to the platelet micro-nano motor suspension containing the polydopamine coating is 1:1-5:1, wherein the platelet density is 1× 10 7-8 /mL, mixed with the platelet micro-nanomotor suspension containing polydopamine coating for 2-12 hours at room temperature.
本发明所述的血小板微纳米马达在制备用于防止癌症的转移和复发药物中的应用。The application of the platelet micro-nano motor of the present invention in the preparation of a medicament for preventing the metastasis and recurrence of cancer.
其中，所述血小板载药微纳米马达可以自主趋向手术切除后的伤口，受到炎性环境或肿瘤微环境刺激，活化释放纳米尺寸的血小板衍生微粒，促进化疗药物如阿霉素释放。施加的近红外激光照射，可以促进血小板衍生微粒被癌细胞摄取，更好地发挥阿霉素的药效。Among them, the platelet-loaded micro-nanomotor can autonomously tend to the wound after surgical resection, and is stimulated by the inflammatory environment or the tumor microenvironment to activate and release nano-sized platelet-derived particles, and promote the release of chemotherapeutic drugs such as doxorubicin. The applied near-infrared laser irradiation can promote the uptake of platelet-derived microparticles by cancer cells, and better exert the efficacy of doxorubicin.
具体而言，本发明利用多巴胺在弱碱性条件下易于自聚合的特点，在血小板表面原位聚合形成聚多巴胺涂层，再利用聚多巴胺丰富的邻苯二醌基团与含胺类药物发生席夫碱反应，或利用聚多巴胺和含胺类化合物或芳香族药物之间的π-π堆积装载药物，形成血小板载药微纳米马达。利用血小板本身的不对称性，在近红外光的照射下，具有光热效应的聚多巴胺涂层可形成不均匀的热泳现象，由此推动血小板载药微纳米马达的运动。Specifically, the present invention utilizes the characteristics of easy self-polymerization of dopamine under weak alkaline conditions to form a polydopamine coating on the platelet surface by in-situ polymerization, and then utilizes the rich o-phthaloquinone group of polydopamine to form a drug containing amines. Schiff base reaction, or using π-π stacking between polydopamine and amine-containing compounds or aromatic drugs to load drugs to form platelet drug-loaded micro-nanomotors. Taking advantage of the asymmetry of platelets, under the irradiation of near-infrared light, the polydopamine coating with photothermal effect can form an uneven thermophoresis phenomenon, thereby promoting the movement of platelet drug-loaded micro-nanomotors.
本发明利用了血小板天然特征，生理条件下的血小板处于静息态，呈圆盘状，在血管壁附近循环。当受到激活信号刺激时，血小板发生黏附、激活和聚集。激活态血小板伸出伪足，呈树突状，同时释放血小板衍生微粒(血小板的质膜)，直径约0.1-1微米。本发明从全血中提取血小板，并在其膜表面修饰聚多巴胺涂层(静息态)，然后添加凝血酶体外激活修饰后的血小板，得到激活态血小板及血小板衍生微粒，这两种产物都来源于原来的血小板，因此它们表面也覆盖有聚多巴胺涂层。而与聚多巴胺结合的药物分子也存在于激活态血小板和血小板衍生微粒表面，形成纳米级的血小板衍生微粒携带药物，比微米级的细胞更有利于药物的组织渗透。The invention utilizes the natural characteristics of platelets, and the platelets under physiological conditions are in a resting state, in the shape of a disc, and circulate near the blood vessel wall. When stimulated by activating signals, platelets adhere, activate and aggregate. Activated platelets protrude pseudopodia, dendritic-like, and release platelet-derived microparticles (the plasma membrane of platelets), about 0.1-1 μm in diameter. The present invention extracts platelets from whole blood, modifies polydopamine coating (resting state) on the membrane surface, and then adds thrombin to activate the modified platelets in vitro to obtain activated platelets and platelet-derived microparticles, both of which are Derived from the original platelets, so they are also coated with polydopamine. The drug molecules combined with polydopamine also exist on the surface of activated platelets and platelet-derived particles, forming nano-scale platelet-derived particles to carry drugs, which are more conducive to the tissue penetration of drugs than micron-sized cells.
有益效果：与现有技术相比，本发明具有以下优点：Beneficial effect: Compared with the prior art, the present invention has the following advantages:
本发明选用天然细胞血小板和生物相容性良好的多巴胺为原料制备血小板载药微纳米马达。该方法原料简单、反应条件温和、操作简便，制得的材料具有良好的生物相容性，可以自主趋向炎症或伤口部位，并通过血小板细胞活化产生血小板衍生微粒，从而释放药物。本发明制备的血小板载药微纳米马达与现有的细胞基微纳米马达相比，本发明提供的血小板载药微纳米马达具有刺激响应性形貌变化，激活后可以产生纳米级衍生微粒，与现有的活细胞给药体系相比(如：巨噬细胞、精细胞、细菌等)克服了当前生物类微米马达给药体系本身尺寸较大的不足，更有利于药物在组织内的渗透，同时利用血小板本身的不对称性，在近红外光的照射下，具有光热效应的聚多巴胺涂层可形成不均匀的热泳现象，由此推动血小板载药微纳米马达的运动，微纳米马达的运动性能可以增强细胞摄取和组织渗透，从而提高药物递送效率。The invention selects natural cell platelets and dopamine with good biocompatibility as raw materials to prepare platelet drug-loaded micro-nano motors. The method has simple raw materials, mild reaction conditions and simple operation, and the prepared material has good biocompatibility, can autonomously tend to inflammation or wound sites, and activate platelet cells to generate platelet-derived particles, thereby releasing drugs. Compared with the existing cell-based micro-nano motor, the platelet drug-loading micro-nano motor prepared by the present invention has a stimuli-responsive morphology change, and can generate nano-scale derivative particles after activation, which is compatible with the Compared with the existing live cell drug delivery systems (such as macrophages, sperm cells, bacteria, etc.), it overcomes the shortcomings of the current biological micro-motor drug delivery system itself, which is relatively large in size, and is more conducive to the penetration of drugs in tissues. At the same time, using the asymmetry of platelets, under the irradiation of near-infrared light, the polydopamine coating with photothermal effect can form a non-uniform thermophoresis phenomenon, thereby promoting the movement of the platelet drug-loaded micro-nano motor, and the micro-nano motor. Kinetic properties can enhance cellular uptake and tissue penetration, thereby increasing drug delivery efficiency.
本发明制得的血小板载药微纳米马达可以利用血小板固有的趋化性自主趋向炎症、血栓、术后创口等病理环境的组织和细胞，再通过微纳米马达的运动可以增强细胞摄取和组织渗透，从而提高药物递送效率，可以有效用于制备用于防止癌症的转移和复发药物中，在生物医学领域具有广阔的应用前景。The platelet drug-loaded micro-nano motor prepared by the invention can utilize the inherent chemotaxis of platelets to autonomously tend to tissues and cells in pathological environments such as inflammation, thrombosis, postoperative wounds, etc., and then through the movement of the micro-nano motor, cell uptake and tissue penetration can be enhanced , thereby improving the drug delivery efficiency, can be effectively used in the preparation of drugs for preventing cancer metastasis and recurrence, and has broad application prospects in the field of biomedicine.
附图说明Description of drawings
图1为血小板的透射电镜图；Figure 1 is a transmission electron microscope image of platelets;
图2为血小板微纳米马达的透射电镜图；Figure 2 is a transmission electron microscope image of the platelet micro-nanomotor;
图3为负载阿霉素的血小板载药微纳米马达的免疫荧光图像；Figure 3 is an immunofluorescence image of the platelet drug-loaded micro-nanomotor loaded with doxorubicin;
图4为近红外激光(808nm，2.5W/cm2)照射下负载阿霉素的血小板载药微纳米马达运动轨迹(5s)；Figure 4 shows the trajectory (5s) of the platelet drug-loaded micro-nanomotor loaded with doxorubicin under the irradiation of near-infrared laser (808nm, 2.5W/cm 2 );
图5为不同材料与MCF-7细胞共孵育24小时后的细胞存活率。Figure 5 shows the cell viability after co-incubating different materials with MCF-7 cells for 24 hours.
具体实施方式Detailed ways
下面通过具体实施例对本发明所述的技术方案给予进一步详细的说明，但有必要提出，以下实施例只用于对发明内容的描述，并不构成对本发明保护范围的限制。The technical solutions of the present invention will be further described in detail below through specific examples, but it is necessary to mention that the following examples are only used to describe the content of the invention, and do not constitute a limitation on the protection scope of the present invention.
实施例1Example 1
一种血小板载药微纳米马达的制备方法，具体包括以下步骤：A preparation method of a platelet drug-loaded micro-nano motor, which specifically comprises the following steps:
(1)血小板微纳米马达的制备：(1) Preparation of platelet micro-nanomotors:
采集新鲜兔全血，1500rpm离心15分钟，小心吸取收集上层富血小板血浆；3000rpm离心10分钟，弃去上层血浆，将沉淀血小板缓慢重悬于PBS(pH＝7.4，下同)缓冲溶液，得到血小板悬浊液。血小板的透射电镜图如图1所示，从图1可以看出血小板呈圆盘状。称取10mg盐酸多巴胺，溶于10mL，10mM Tris-HCl缓冲液(pH＝8.5)。通过细胞计数法调节血小板悬浊液中血小板密度为1×108个/mL，取前述盐酸多巴胺溶液1mL与1mL(1×108个)血小板悬浊液混合，置于静音混合器，室温下聚合2小时，得到含有聚多巴胺涂层的血小板微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。图2为血小板微纳米马达的透射电镜图，从图2可以看出聚多巴胺包裹在圆盘状血小板外周，其厚度约为0.3-1μm，其外圈颜色较浅的为聚多巴胺涂层厚度约为0.1-1μm。Collect fresh rabbit whole blood, centrifuge at 1500 rpm for 15 minutes, carefully draw and collect the upper platelet-rich plasma; centrifuge at 3000 rpm for 10 minutes, discard the upper plasma, and slowly resuspend the precipitated platelets in PBS (pH=7.4, the same below) buffer solution to obtain platelets suspension. The TEM image of the platelets is shown in Figure 1, and it can be seen from Figure 1 that the platelets are disc-shaped. 10 mg of dopamine hydrochloride was weighed and dissolved in 10 mL of 10 mM Tris-HCl buffer (pH=8.5). The platelet density in the platelet suspension was adjusted to 1 × 10 8 /mL by the cell counting method, and 1 mL of the aforementioned dopamine hydrochloride solution was mixed with 1 mL (1 × 10 8 ) of the platelet suspension, and placed in a silent mixer at room temperature. After polymerization for 2 hours, a platelet micro-nanomotor suspension containing polydopamine coating was obtained, centrifuged at 3000 rpm for 10 min to remove the lower layer precipitate, added with PBS buffer and mixed, centrifuged and washed, and dispersed in PBS buffer for use. Figure 2 is a transmission electron microscope image of the platelet micro-nanomotor. It can be seen from Figure 2 that polydopamine is wrapped around the disk-shaped platelet with a thickness of about 0.3-1 μm. 0.1-1μm.
(2)血小板载药微纳米马达的制备：(2) Preparation of platelet drug-loaded micro-nanomotors:
称取10mg盐酸阿霉素，超声溶解于10mL PBS缓冲液(pH＝7.4，下同)。通过细胞计数法调节(1)中含有聚多巴胺涂层的血小板微纳米马达悬浊液中血小板的密度为1×108个/mL。取前述盐酸阿霉素溶液4mL与1mL含有聚多巴胺涂层的血小板微纳米马达悬浊液(1×108个)混合，置于静音混合器，室温下避光反应2小时，得到负载阿霉素的血小板载药微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。将所得负载阿霉素的血小板载药微纳米马达与DiO染料共孵育，标记血小板膜并置于共聚焦显微镜下观察，其免疫荧光图像如图3所示。从图3可以看出红色荧光(阿霉素DOX，中图)与绿色荧光(血小板PLTs，左图)几乎重合(右图)，证明了药物的成功负载。将所得的负载阿霉素的血小板载药微纳米马达在近红外激光(808nm，2.5W/cm2)照射下的运动视频截图如图4所示，黑色细线代表血小板载药微纳米马达在5s内的运动轨迹，证明血小板载药微纳米马达在近红外激光的照射下可以运动。10 mg of doxorubicin hydrochloride was weighed and dissolved in 10 mL of PBS buffer (pH=7.4, the same below) by ultrasonication. The density of platelets in the platelet micro-nanomotor suspension containing the polydopamine coating in (1) was adjusted to 1×10 8 /mL by cytometry. Mix 4 mL of the aforementioned doxorubicin hydrochloride solution with 1 mL of the platelet micro-nano motor suspension containing polydopamine coating (1 × 10 8 ), place it in a silent mixer, and react at room temperature for 2 hours in the dark to obtain a loaded doxorubicin The platelet drug-loaded micro-nano motor suspension was prepared by centrifugation at 3000 rpm for 10 min to remove the lower layer of sediment, and PBS buffer was added to mix evenly. After centrifugal washing, it was dispersed in PBS buffer for use. The obtained doxorubicin-loaded platelet drug-loaded micro-nanomotor was co-incubated with DiO dye, and the platelet membrane was labeled and observed under a confocal microscope. The immunofluorescence image was shown in Figure 3. From Figure 3, it can be seen that the red fluorescence (doxorubicin DOX, middle panel) almost coincides with the green fluorescence (platelet PLTs, left panel) (right panel), demonstrating the successful loading of the drug. A screenshot of the motion video of the obtained doxorubicin-loaded platelet drug-loaded micro-nanomotor under near-infrared laser (808nm, 2.5W/cm 2 ) irradiation is shown in Figure 4. The black thin line represents the platelet-loaded drug-loaded micro-nanomotor in the The trajectory within 5s proves that the platelet drug-loaded micro-nanomotor can move under the irradiation of near-infrared laser.
将0.1mL细胞密度为5×104个/mL的人乳腺癌细胞(MCF-7)接种于96孔板，置于37℃、5％CO2培养箱中培养12h。然后吸出培养基，实验组每孔分别加入0.2mL(1×108个/mL)血小板(PLTs)或上述血小板载药微纳米马达或新鲜培养基，加入新鲜培养基的组别每孔再用近红外激光器(808nm，2.5W/cm2)照射10min，空白组(Control)每孔加入等量新鲜细胞培养基作对照，孵育24h后采用MTT法测试细胞存活率。结果如图5所示，实验组细胞存活率均大于90％，表明本研究制备的血小板微纳米马达具有良好的生物相容性。0.1 mL of human breast cancer cells (MCF-7) with a cell density of 5×10 4 cells/mL were seeded in a 96-well plate, and cultured in a 37° C., 5% CO 2 incubator for 12 h. Then the medium was aspirated, and 0.2 mL (1×10 8 /mL) platelets (PLTs) or the above-mentioned platelet-loaded micro-nanomotors or fresh medium were added to each well of the experimental group. The near-infrared laser (808nm, 2.5W/cm 2 ) was irradiated for 10min, and the blank group (Control) was added with an equal amount of fresh cell culture medium to each well as a control, and the cell viability was tested by MTT method after incubation for 24h. The results are shown in Figure 5. The cell survival rates of the experimental groups were all greater than 90%, indicating that the platelet micro-nanomotors prepared in this study have good biocompatibility.
实施例2Example 2
一种血小板载药微纳米马达的制备方法，具体包括以下步骤：A preparation method of a platelet drug-loaded micro-nano motor, which specifically includes the following steps:
(1)血小板微纳米马达的制备：(1) Preparation of platelet micro-nanomotors:
按照实施例1所述梯度离心法分离提取血小板，得血小板悬浊液。通过细胞计数法调节血小板悬浊液中血小板密度为1×107个/mL。称取20mg盐酸多巴胺，溶于10mL，10mMTris-HCl缓冲液(pH＝8.5)，取出1mL与10mL(1×107个)血小板悬浊液混合，置于静音混合器，室温下聚合2小时，得到血小板微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。The platelets were separated and extracted according to the gradient centrifugation method described in Example 1 to obtain a platelet suspension. The platelet density in the platelet suspension was adjusted to 1×10 7 /mL by cell counting. Weigh 20 mg of dopamine hydrochloride, dissolve it in 10 mL, 10 mM Tris-HCl buffer (pH=8.5), take out 1 mL and mix it with 10 mL (1×10 7 ) platelet suspension, put it in a silent mixer, and polymerize at room temperature for 2 hours. The platelet micro-nano motor suspension was obtained, centrifuged at 3000 rpm for 10 min to remove the lower layer precipitate, added PBS buffer to mix well, centrifuged and washed, and dispersed in PBS buffer for use.
(2)血小板载药微纳米马达的制备：(2) Preparation of platelet drug-loaded micro-nanomotors:
称取10mg盐酸阿霉素，超声溶解于10mL PBS缓冲液。通过细胞计数法调节(1)中含有聚多巴胺涂层的血小板微纳米马达悬浊液中血小板的密度为1×107个/mL。移取前述盐酸阿霉素溶液1mL与1mL含有聚多巴胺涂层的血小板微纳米马达悬浊液混合(1×107个)，置于静音混合器，室温下避光反应2小时，得到负载阿霉素的血小板载药微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。Weigh 10 mg of doxorubicin hydrochloride and dissolve it in 10 mL of PBS buffer by sonication. The density of platelets in the platelet micro-nanomotor suspension containing the polydopamine coating in (1) was adjusted to 1×10 7 /mL by cytometry. Pipette 1 mL of the aforementioned doxorubicin hydrochloride solution and mix it with 1 mL of the platelet micro-nano motor suspension containing polydopamine coating (1×10 7 pieces), place it in a silent mixer, and react at room temperature in the dark for 2 hours to obtain the loaded doxorubicin. The platelet drug-loaded micro-nanomotor suspension of tetracycline was centrifuged at 3000 rpm for 10 min to remove the lower layer of the precipitate, and PBS buffer was added to mix evenly. After centrifugal washing, it was dispersed in PBS buffer for use.
实施例3Example 3
一种血小板载药微纳米马达的制备方法，具体包括以下步骤：A preparation method of a platelet drug-loaded micro-nano motor, which specifically comprises the following steps:
(1)血小板微纳米马达的制备：(1) Preparation of platelet micro-nanomotors:
采集新鲜兔全血，1000rpm离心20分钟，小心吸取收集上层富血小板血浆；2500rpm离心15分钟，弃去上层血浆，将沉淀血小板缓慢重悬于PBS缓冲溶液，得到血小板悬浊液。通过细胞计数法调节血小板悬浊液中血小板密度为1×108个/mL。称取1mg盐酸多巴胺，溶于10mL，10mM Tris-HCl缓冲液(pH＝8.5)，取出1mL与1mL(1×108个)血小板悬浊液混合，置于静音混合器，室温下聚合6小时，得到血小板微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。Fresh rabbit whole blood was collected, centrifuged at 1000rpm for 20 minutes, and the upper layer of platelet-rich plasma was carefully collected; centrifuged at 2500rpm for 15 minutes, the upper layer of plasma was discarded, and the precipitated platelets were slowly resuspended in PBS buffer solution to obtain a platelet suspension. The platelet density in the platelet suspension was adjusted to 1×10 8 /mL by cell counting. Weigh 1 mg of dopamine hydrochloride, dissolve it in 10 mL, 10 mM Tris-HCl buffer (pH=8.5), take out 1 mL and mix it with 1 mL (1×10 8 ) platelet suspension, put it in a silent mixer, and polymerize at room temperature for 6 hours , to obtain a platelet micro-nano motor suspension, centrifuged at 3000 rpm for 10 min to remove the lower layer of the precipitate, added PBS buffer to mix well, centrifuged and washed, and dispersed in PBS buffer for use.
(2)血小板载药微纳米马达的制备：(2) Preparation of platelet drug-loaded micro-nanomotors:
称取4mg盐酸尼莫司汀，溶解于4mL PBS缓冲液。通过细胞计数法调节(1)中含有聚多巴胺涂层的血小板微纳米马达悬浊液中血小板的密度为1×108个/mL，取1mL与前述4mL盐酸尼莫司汀溶液混合，室温下避光反应2小时，得负载尼莫司汀的血小板载药马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液保存备用。Weigh 4 mg of nimustine hydrochloride and dissolve it in 4 mL of PBS buffer. The density of platelets in the platelet micro-nanomotor suspension containing polydopamine coating in (1) was adjusted to 1×10 8 /mL by cytometry, and 1 mL was mixed with the aforementioned 4 mL of nimustine hydrochloride solution, and at room temperature The reaction was performed in the dark for 2 hours to obtain a nimustine-loaded platelet drug-loaded motor suspension, centrifuged at 3000 rpm for 10 min to remove the lower layer precipitate, added with PBS buffer and mixed, and dispersed in PBS buffer after centrifugation and washed for later use.
实施例4Example 4
一种血小板载药微纳米马达的制备方法，具体包括以下步骤：A preparation method of a platelet drug-loaded micro-nano motor, which specifically includes the following steps:
(1)血小板微纳米马达的制备：(1) Preparation of platelet micro-nanomotors:
按照实施例1所述梯度离心法分离提取血小板，得血小板悬浊液。通过细胞计数法调节血小板悬浊液中血小板密度为1×108个/mL。称取5mg盐酸多巴胺，溶于10mL，10mMTris-HCl缓冲液(pH＝8.5)，取出1mL与1mL(1×108个)血小板悬浊液混合，置于静音混合器，室温下聚合2小时，得到血小板微纳米马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液备用。The platelets were separated and extracted according to the gradient centrifugation method described in Example 1 to obtain a platelet suspension. The platelet density in the platelet suspension was adjusted to 1×10 8 /mL by cell counting. Weigh 5 mg of dopamine hydrochloride, dissolve it in 10 mL, 10 mM Tris-HCl buffer (pH=8.5), take out 1 mL and mix it with 1 mL (1×10 8 ) platelet suspension, put it in a silent mixer, and polymerize at room temperature for 2 hours. The platelet micro-nano motor suspension was obtained, centrifuged at 3000 rpm for 10 min to remove the lower layer precipitate, added PBS buffer to mix well, centrifuged and washed, and dispersed in PBS buffer for use.
(2)血小板载药微纳米马达的制备：(2) Preparation of platelet drug-loaded micro-nanomotors:
称取2mg丝裂霉素，溶解于2mL PBS缓冲液。通过细胞计数法调节(1)中含有聚多巴胺涂层的血小板微纳米马达悬浊液中血小板的密度为1×108个/mL，取1mL与前述2mL丝裂霉素溶液混合，室温下避光反应2小时，得负载丝裂霉素的血小板载药马达悬浊液，3000rpm离心10min取下层沉淀，加入PBS缓冲液混匀，离心洗涤后分散于PBS缓冲液保存备用。Weigh 2 mg of mitomycin and dissolve it in 2 mL of PBS buffer. The density of platelets in the platelet micro-nanomotor suspension containing polydopamine coating in (1) was adjusted to 1 × 10 8 /mL by cytometry, and 1 mL was mixed with the aforementioned 2 mL of mitomycin solution, and kept at room temperature. After photoreaction for 2 hours, a mitomycin-loaded platelet drug-loaded motor suspension was obtained, centrifuged at 3000 rpm for 10 min to remove the lower sediment, added with PBS buffer to mix well, centrifuged and washed, and dispersed in PBS buffer for later use.
实施例5Example 5
实施例5与实施例1制备方法相同，不同之处在于，盐酸阿霉素药物浓度为0.1mg/mL，与含有聚多巴胺涂层的血小板微纳米马达悬浊液的体积比为4:1，其中血小板密度为1×107个/mL，在室温下混合4小时。The preparation method of Example 5 is the same as that of Example 1, except that the drug concentration of doxorubicin hydrochloride is 0.1 mg/mL, and the volume ratio to the platelet micro-nano motor suspension containing the polydopamine coating is 4:1, The platelet density was 1×10 7 /mL, and the mixture was mixed at room temperature for 4 hours.
实施例6Example 6
实施例6与实施例1制备方法相同，不同之处在于，盐酸阿霉素药物浓度为2mg/mL，与含有聚多巴胺涂层的血小板微纳米马达悬浊液的体积比为5:1，其中血小板密度为1×107个/mL，在室温下混合12小时。The preparation method of Example 6 is the same as that of Example 1, except that the drug concentration of doxorubicin hydrochloride is 2 mg/mL, and the volume ratio with the platelet micro-nano motor suspension containing polydopamine coating is 5:1, wherein The platelet density was 1×10 7 /mL, and the mixture was mixed at room temperature for 12 hours.
本发明实施例制备的血小板微纳米马达在制备用于防止癌症的转移和复发药物中，血小板载药微纳米马达可以自主趋向手术切除后的伤口，受到炎性环境或肿瘤微环境刺激，活化释放纳米尺寸的血小板衍生微粒，促进化疗药物如阿霉素释放。本发明通过施加的近红外激光照射，利用血小板本身的不对称性，在近红外光的照射下，具有光热效应的聚多巴胺涂层可形成不均匀的热泳现象，由此推动血小板载药微纳米马达的运动，可以促进血小板衍生微粒被癌细胞摄取，更好地发挥如阿霉素、尼莫司汀或丝裂霉素的药效。The platelet micro-nanomotor prepared in the embodiment of the present invention is used for the preparation of a drug for preventing cancer metastasis and recurrence. The platelet drug-loaded micro-nanomotor can autonomously tend to the wound after surgical resection, and is stimulated by the inflammatory environment or the tumor microenvironment to activate and release Nano-sized platelet-derived microparticles that facilitate the release of chemotherapeutic drugs such as doxorubicin. The present invention utilizes the asymmetry of the platelet itself through the applied near-infrared laser irradiation, and under the irradiation of near-infrared light, the polydopamine coating with photothermal effect can form an uneven thermophoresis phenomenon, thereby promoting the platelet drug-loaded microparticles. The movement of nanomotors can promote the uptake of platelet-derived microparticles by cancer cells to better exert the efficacy of drugs such as doxorubicin, nimustine or mitomycin.
Claims (8)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010509926.9A CN111632038B (en) | 2020-06-05 | 2020-06-05 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010509926.9A CN111632038B (en) | 2020-06-05 | 2020-06-05 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111632038A true CN111632038A (en) | 2020-09-08 |
| CN111632038B CN111632038B (en) | 2022-02-11 |
Family
ID=72322792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010509926.9A Active CN111632038B (en) | 2020-06-05 | 2020-06-05 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111632038B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113633788A (en) * | 2021-08-02 | 2021-11-12 | 郑州大学第一附属医院 | A kind of preparation method of high-adhesion drug sustained-release body that can be developed for nerve block |
| CN114425043A (en) * | 2021-12-08 | 2022-05-03 | 深圳先进技术研究院 | A macrophage-based living cell drug delivery system and its preparation method and application |
| CN116159152A (en) * | 2022-12-27 | 2023-05-26 | 中国医学科学院放射医学研究所 | Synthesis method of platelet nanomotor loaded with gold nanoparticles in situ |
| CN117982987A (en) * | 2024-04-03 | 2024-05-07 | 四川厚浦生物科技有限公司 | Leukocyte filtering material and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233302A (en) * | 2017-07-25 | 2017-10-10 | 中国科学院青岛生物能源与过程研究所 | A kind of preparation method of nano-cellulose/poly-dopamine composite intelligent gel medicine slow-release material |
| US20180087116A1 (en) * | 2017-02-11 | 2018-03-29 | Mahmoud Amouzadeh Tabrizi | Methods and biosensors for tumor detection |
| CN108042802A (en) * | 2017-12-11 | 2018-05-18 | 哈尔滨工业大学 | A kind of artificial red cells motor and preparation method thereof |
| CN109091673A (en) * | 2018-09-11 | 2018-12-28 | 浙江理工大学 | It is a kind of integrate targeting, photo-thermal red blood cell biomimetic type nanoparticle preparation method |
| CN110251479A (en) * | 2019-06-06 | 2019-09-20 | 南京师范大学 | A red blood cell membrane-wrapped bionic blood hexavalent chromium reduction remover/magnetic nanomotor and its preparation method and application |
-
2020
- 2020-06-05 CN CN202010509926.9A patent/CN111632038B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180087116A1 (en) * | 2017-02-11 | 2018-03-29 | Mahmoud Amouzadeh Tabrizi | Methods and biosensors for tumor detection |
| CN107233302A (en) * | 2017-07-25 | 2017-10-10 | 中国科学院青岛生物能源与过程研究所 | A kind of preparation method of nano-cellulose/poly-dopamine composite intelligent gel medicine slow-release material |
| CN108042802A (en) * | 2017-12-11 | 2018-05-18 | 哈尔滨工业大学 | A kind of artificial red cells motor and preparation method thereof |
| CN109091673A (en) * | 2018-09-11 | 2018-12-28 | 浙江理工大学 | It is a kind of integrate targeting, photo-thermal red blood cell biomimetic type nanoparticle preparation method |
| CN110251479A (en) * | 2019-06-06 | 2019-09-20 | 南京师范大学 | A red blood cell membrane-wrapped bionic blood hexavalent chromium reduction remover/magnetic nanomotor and its preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| MIMI WAN ET AL.: "Platelet-derived porous nanomotor for thrombus therapy", 《SCIENCE ADVANCES》 * |
| MIMI WAN ET AL.: "Systematic Research and Evaluation Models of Nanomotors for Cancer Combined Therapy", 《ANGEWANDTE》 * |
| 吴志光等: "红细胞伪装微纳米马达", 《中国化学会第十五届胶体与界面化学会议论文集（第二分会）》 * |
| 孔磊等: "自驱动微纳米马达的设计原理与结构简化方法", 《科学通报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113633788A (en) * | 2021-08-02 | 2021-11-12 | 郑州大学第一附属医院 | A kind of preparation method of high-adhesion drug sustained-release body that can be developed for nerve block |
| CN114425043A (en) * | 2021-12-08 | 2022-05-03 | 深圳先进技术研究院 | A macrophage-based living cell drug delivery system and its preparation method and application |
| CN116159152A (en) * | 2022-12-27 | 2023-05-26 | 中国医学科学院放射医学研究所 | Synthesis method of platelet nanomotor loaded with gold nanoparticles in situ |
| CN117982987A (en) * | 2024-04-03 | 2024-05-07 | 四川厚浦生物科技有限公司 | Leukocyte filtering material and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111632038B (en) | 2022-02-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111632038A (en) | A kind of platelet drug-loaded micro-nano motor and its preparation method and application | |
| Bigham et al. | MOFs and MOF‐based composites as next‐generation materials for wound healing and dressings | |
| Feng et al. | The effect of nanofibrous galactosylated chitosan scaffolds on the formation of rat primary hepatocyte aggregates and the maintenance of liver function | |
| CN111265533A (en) | Preparation method of core-shell nanoparticles based on lipid membrane and metal organic framework | |
| Zhang et al. | Porous electrospun fibers with self‐sealing functionality: An enabling strategy for trapping biomacromolecules | |
| Wu et al. | Mucus-permeable polymyxin B-hyaluronic acid/poly (lactic-co-glycolic acid) nanoparticle platform for the nebulized treatment of lung infections | |
| Cao et al. | Multifunctional photo-and magnetoresponsive graphene oxide–Fe3O4 nanocomposite–alginate hydrogel platform for ice recrystallization inhibition | |
| CN108553650B (en) | A biomimetic nano-red blood cell gene carrier and its preparation method and application | |
| Kim et al. | Macromolecular Engineering of Thermoresponsive Metal–Phenolic Networks | |
| CN101259097A (en) | A magnetic tumor dual-targeting polymer nanomicelle and its preparation method | |
| CN110292652B (en) | Mercaptophenylboronic acid-activated gold nanoparticles, preparation method and application thereof | |
| CN101390826A (en) | A magnetic tumor targeting polymer nanovesicle and its preparation method | |
| CN113633625A (en) | A kind of nano-medicine carrying oxidative phosphorylation inhibitor on heteromembrane and preparation method thereof | |
| CN105456233B (en) | A kind of long-acting slow-release antibacterial film of high drug load and preparation method thereof | |
| Chen et al. | Photothermal Nanozyme‐Encapsulating Microneedles for Synergistic Treatment of Infected Wounds | |
| CN115192708B (en) | Nanocomposite loaded with antitumor drug, nano drug-carrying system, preparation and application | |
| Bai et al. | Blood cellular membrane-coated Au/polydopamine nanoparticle-targeted NIR-II antibacterial therapy | |
| Li et al. | Development of magnetic poly (L-lactic Acid) nanofibrous microspheres for transporting and delivering targeted cells | |
| CN114796153B (en) | A preparation method of drug-loaded degradable molecularly imprinted polymer nanoparticles | |
| CN113999410B (en) | Preparation method of double-crosslinked antibacterial hydrogel for repairing burn and scald wound | |
| CN108743971B (en) | Preparation method and application of drug-loaded polypyrrole nanoparticles | |
| CN114796151A (en) | Nano-drug hydrogel interpenetrating network system microcapsule with targeted tumor microenvironment controlled release | |
| Wang et al. | Reprogrammed Lung Metastasis Immunodeficiency via Targeted Penetrated Delivery of M1 Macrophage‐Wrapped NanoCubes‐Mediated T Cell Infiltration | |
| CN117447663A (en) | Porphyrin-based conjugated organic framework material based on membrane-penetrating peptide modification, and preparation method and application thereof | |
| CN114642742B (en) | Multiple stimulus response nano material-polymer composite hydrogel and preparation method 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 |