CN110237268A - 一种载有阿霉素的双响应脂质体微泡复合物的制备方法 - Google Patents
一种载有阿霉素的双响应脂质体微泡复合物的制备方法 Download PDFInfo
- Publication number
- CN110237268A CN110237268A CN201910648607.3A CN201910648607A CN110237268A CN 110237268 A CN110237268 A CN 110237268A CN 201910648607 A CN201910648607 A CN 201910648607A CN 110237268 A CN110237268 A CN 110237268A
- Authority
- CN
- China
- Prior art keywords
- crgd
- peg2000
- dspe
- liposome
- microbubble
- 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
- 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 title claims abstract description 86
- 239000002502 liposome Substances 0.000 title claims abstract description 79
- 150000001875 compounds Chemical class 0.000 title claims abstract description 61
- 229940009456 adriamycin Drugs 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 108010045325 cyclic arginine-glycine-aspartic acid peptide Proteins 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 26
- 229960004679 doxorubicin Drugs 0.000 claims abstract description 21
- 230000008685 targeting Effects 0.000 claims abstract description 21
- 150000002632 lipids Chemical class 0.000 claims abstract description 20
- 239000008187 granular material Substances 0.000 claims abstract description 10
- 229960002685 biotin Drugs 0.000 claims abstract description 9
- 239000011616 biotin Substances 0.000 claims abstract description 9
- 230000006287 biotinylation Effects 0.000 claims abstract description 8
- 238000007413 biotinylation Methods 0.000 claims abstract description 8
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-N 0.000 claims abstract description 7
- TYAQXZHDAGZOEO-KXQOOQHDSA-N 1-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCC TYAQXZHDAGZOEO-KXQOOQHDSA-N 0.000 claims abstract description 7
- 208000014446 corneal intraepithelial dyskeratosis-palmoplantar hyperkeratosis-laryngeal dyskeratosis syndrome Diseases 0.000 claims abstract description 7
- 108090001008 Avidin Proteins 0.000 claims abstract description 6
- 238000006703 hydration reaction Methods 0.000 claims abstract description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 18
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 8
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 238000002390 rotary evaporation Methods 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 4
- 239000007979 citrate buffer Substances 0.000 claims description 3
- 230000036571 hydration Effects 0.000 claims description 3
- 239000002105 nanoparticle Substances 0.000 claims description 3
- 150000003904 phospholipids Chemical class 0.000 claims description 3
- GDIYMWAMJKRXRE-UHFFFAOYSA-N (2z)-2-[(2e)-2-[2-chloro-3-[(z)-2-(1,3,3-trimethylindol-1-ium-2-yl)ethenyl]cyclohex-2-en-1-ylidene]ethylidene]-1,3,3-trimethylindole Chemical compound CC1(C)C2=CC=CC=C2N(C)C1=CC=C1C(Cl)=C(C=CC=2C(C3=CC=CC=C3[N+]=2C)(C)C)CCC1 GDIYMWAMJKRXRE-UHFFFAOYSA-N 0.000 claims description 2
- 239000004698 Polyethylene Substances 0.000 claims description 2
- 239000002202 Polyethylene glycol Substances 0.000 claims description 2
- 229920005654 Sephadex Polymers 0.000 claims description 2
- 239000012507 Sephadex™ Substances 0.000 claims description 2
- 239000003054 catalyst Substances 0.000 claims description 2
- 238000004440 column chromatography Methods 0.000 claims description 2
- 125000004122 cyclic group Chemical group 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 2
- 229920000573 polyethylene Polymers 0.000 claims description 2
- 229920001223 polyethylene glycol Polymers 0.000 claims description 2
- 229960002317 succinimide Drugs 0.000 claims description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims 2
- -1 Acyl ethanol Chemical compound 0.000 claims 1
- BYWUNUDERLAEFZ-UHFFFAOYSA-N C(CCCCCCCCCCCCCCC)[P] Chemical compound C(CCCCCCCCCCCCCCC)[P] BYWUNUDERLAEFZ-UHFFFAOYSA-N 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N EtOH Substances CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 claims 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 description 53
- 239000003814 drug Substances 0.000 description 18
- 230000000694 effects Effects 0.000 description 17
- 238000011580 nude mouse model Methods 0.000 description 17
- 241000699660 Mus musculus Species 0.000 description 15
- 229940079593 drug Drugs 0.000 description 15
- 238000000015 thermotherapy Methods 0.000 description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 239000000523 sample Substances 0.000 description 8
- 239000013558 reference substance Substances 0.000 description 7
- 239000007924 injection Substances 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 206010006187 Breast cancer Diseases 0.000 description 4
- 208000026310 Breast neoplasm Diseases 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 230000021164 cell adhesion Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 230000005909 tumor killing Effects 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- ATHHXGZTWNVVOU-UHFFFAOYSA-N N-methylformamide Chemical compound CNC=O ATHHXGZTWNVVOU-UHFFFAOYSA-N 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 210000003716 mesoderm Anatomy 0.000 description 2
- 238000000879 optical micrograph Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000003672 processing method Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000010183 spectrum analysis Methods 0.000 description 2
- JGVWCANSWKRBCS-UHFFFAOYSA-N tetramethylrhodamine thiocyanate Chemical compound [Cl-].C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=C(SC#N)C=C1C(O)=O JGVWCANSWKRBCS-UHFFFAOYSA-N 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- IRPKBYJYVJOQHQ-UHFFFAOYSA-M (2e)-2-[(2e)-2-[2-chloro-3-[(e)-2-(3,3-dimethyl-1-propylindol-1-ium-2-yl)ethenyl]cyclohex-2-en-1-ylidene]ethylidene]-3,3-dimethyl-1-propylindole;iodide Chemical compound [I-].CC1(C)C2=CC=CC=C2N(CCC)\C1=C\C=C/1C(Cl)=C(\C=C/C=2C(C3=CC=CC=C3[N+]=2CCC)(C)C)CCC\1 IRPKBYJYVJOQHQ-UHFFFAOYSA-M 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 208000003170 Bronchiolo-Alveolar Adenocarcinoma Diseases 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 206010020843 Hyperthermia Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 208000021712 Soft tissue sarcoma Diseases 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 241000256856 Vespidae Species 0.000 description 1
- 208000008383 Wilms tumor Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 230000003444 anaesthetic effect Effects 0.000 description 1
- 150000001454 anthracenes Chemical class 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 239000003972 antineoplastic antibiotic Substances 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000006793 arrhythmia Effects 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 210000003038 endothelium Anatomy 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 230000036031 hyperthermia Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000022013 kidney Wilms tumor Diseases 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000013532 laser treatment Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- 230000003680 myocardial damage Effects 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- DPBLXKKOBLCELK-UHFFFAOYSA-N n-pentylamine Natural products CCCCCN DPBLXKKOBLCELK-UHFFFAOYSA-N 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 201000008026 nephroblastoma Diseases 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229940100684 pentylamine Drugs 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 210000002706 plastid Anatomy 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Classifications
-
- 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
- A61K41/0047—Sonopheresis, i.e. ultrasonically-enhanced transdermal delivery, electroporation of a pharmacologically active agent
-
- 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
- A61K47/62—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 a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- 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
- A61K47/6905—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 colloid or an emulsion
- A61K47/6911—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 colloid or an emulsion the form being a liposome
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
- A61K49/0089—Particulate, powder, adsorbate, bead, sphere
- A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
-
- 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
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
- A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
-
- 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)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Dermatology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种载有阿霉素的双响应脂质体微泡复合物的制备方法,该法由以下步骤组成:(1)将DSPE‑PEG2000‑NHS与cRGD反应得到DSPE‑PEG2000‑cRGD;(2)将DPPC、MSPC、DSPE‑PEG2000‑cRGD和DSPE‑PEG2000‑Biotin水化反应,得到空载携cRGD肽的靶向温敏脂质体纳米粒;(3)用靶向温敏脂质体纳米粒携带阿霉素得到携cRGD肽靶向阿霉素温敏脂质体;(4)将IR780溶液加入到生物素化脂质微泡中;(5)先取所述载IR780脂质微泡加入亲和素孵育,再加入携cRGD肽靶向阿霉素温敏脂质体即得所述的双响应脂质体微泡复合物。本方法所得到的复合物既可减少阿霉素在循环中的损失,又可使阿霉素集中分布于肿瘤中心部位。
Description
技术领域
本发明涉及以所用的非有效成分为特征的医用配制品,具体涉及载有阿霉素的抗肿瘤控释药物。
技术背景
阿霉素是一种蒽环类抗肿瘤抗生素,可阻止肿瘤细胞的生长,抗瘤谱较广,对多种肿瘤均有作用,属周期非特异性药物,对各种生长周期的肿瘤细胞都有杀灭作用。阿霉素对急性淋巴细胞白血病、急性髓细胞性白血病、肾母细胞瘤、神经母细胞瘤,软组织和骨肉瘤、乳腺癌、卵巢癌、膀胱癌,甲状腺癌,胃癌,恶性淋巴瘤和支气管癌都有疗效。目前临床给药多为静脉滴注,但阿霉素不能透过血脑屏障,且静注后该药迅速分布全身,有强烈的毒副作用,主要表现在:心脏毒性,轻者表现为心律失常,重者出现进行性心肌病变而发生充血性心力衰竭;致白细胞和血小板减少,骨髓抑制;毛发脱落;消化道反应,恶心、食欲减退;药物溢出血管外可引起组织溃疡及坏死。这些毒副作用限制了阿霉素在临床化疗的广泛应用,尽管用药剂量很大,能达到病灶部位发挥疗效的比例却很低。为降低其毒性、提高疗效,阿霉素新剂型的研究成为亟待解决的问题。
如何将阿霉素递送至深部肿瘤组织是提高实体肿瘤临床疗效的重要问题。含溶血磷脂温敏脂质体是一种快速释药型阿霉素温敏脂质体制剂,其在预热(41-43℃)的肿瘤血管内可以快速释放大量阿霉素,促使血管内的阿霉素顺浓度梯度弥散进入肿瘤间质,提高肿瘤的药物摄取,可以有效提高阿霉素的抑瘤效果。但是体循环中的一些生物大分子(如血清蛋白)会与温敏脂质体上的溶血磷脂结合,破坏脂质体的稳定性,造成包裹药物的泄露,阻碍药物在肿瘤深部的递送。而以往此类温敏脂质体研究多集中于肿瘤血管内释药的应用,温敏脂质体在间质内释药的应用受到其有效循环时间短的限制一直未有突破。
超声微泡靶向破坏技术(UTMD)是一种具有发展潜力的药物/基因局部靶向递送方法。既往研究表明,微泡表面连接纳米药物形成复合物联合超声辐照可以提高药物递送效率,微泡破裂时产生的瞬态空化作用(微射流、喷射流、瞬间高压)可以使局部的血管内皮间隙增大,血管通透性升高。但以往研究鲜少关注UTMD技术在超声辐照后短时间内纳米药物在肿瘤间质蓄积效应。
发明内容
本发明所要解决的技术问题是提供一种载有阿霉素的双响应脂质体微泡复合物的制备方法,该方法所得到的复合物既可减少阿霉素在循环中的损失,又可使阿霉素集中分布于肿瘤中心部位。
本发明解决上述问题的技术案如下:
一种载有阿霉素的双响应脂质体微泡复合物的制备方法,该法由以下步骤组成:
(1)按DSPE-PEG2000-NHS︰cRGD=1︰2的摩尔比取DSPE-PEG2000-NHS和cRGD溶于二甲基甲酰胺(DMF)中,加入DSPE-PEG2000-NHS和cRGD质量和0.2%的催化剂三乙胺,室温下搅拌反应4h,真空旋转蒸发去除溶剂及三乙胺后得到DSPE-PEG2000-cRGD;其中,所述的DSPE-PEG2000-NHS为分子量2000的二棕榈酰磷脂酰乙醇胺-聚乙二醇-琥珀酰亚胺酯,所述的cRGD为环状低聚肽;
(2)按DPPC︰MSPC︰DSPE-PEG2000-cRGD︰DSPE-PEG2000-Biotin=82︰8︰8︰2的摩尔比取DPPC、MSPC、DSPE-PEG2000-cRGD和DSPE-PEG2000-Biotin溶于氯仿中,常温常压旋转混合均匀;然后,室温下抽真空,旋转蒸发挥去溶剂,并在容器底部形成一层白色磷脂薄膜,真空干燥,加入pH4.0的柠檬酸盐缓冲液水化处理,待磷脂水化完全后,用孔径为100nm滤膜的脂质体挤出器挤压,得到空载携cRGD肽的靶向温敏脂质体纳米粒;其中,所述的DPPC为二棕榈酰磷脂酰胆碱,所述的MSPC为溶血磷脂,所述的DSPE-PEG2000-Biotin为生物素化二棕榈酰磷脂酰乙醇胺-聚乙二醇;
(3)将所得到的靶向温敏脂质体纳米粒的pH值调至7.4,然后按阿霉素︰靶向温敏脂质体纳米粒=1︰20的质量比加入阿霉素(DOX)水溶液,混匀,37℃水浴20min后,用G-50葡聚糖凝胶柱层析,得到携cRGD肽靶向阿霉素温敏脂质体;
(4)取IR-780加入二甲基亚砜(DMSO)配制成浓度为2mg/mL的IR780溶液,然后按IR780溶液︰生物素化脂质微泡=1︰100的体积比将IR780溶液加入到生物素化脂质微泡中,混匀,4℃条件下低速离心,取上层载IR780脂质微泡;其中,所述的IR780为七甲川花菁染料;
(5)按携cRGD肽靶向阿霉素温敏脂质体︰亲和素(Avidin)︰载IR780脂质微泡=5︰1︰5的体积比先取所述载IR780脂质微泡加入亲和素,混匀,冰上孵育30min,然后加入所述携cRGD肽靶向阿霉素温敏脂质体,轻柔混匀后4℃条件下孵育30min,洗涤,得到所述的双响应脂质体微泡复合物。
本方法得到的双响应脂质体微泡复合物较现有技术具有以下有益效果:
联合超声/激光辐照提高含溶血磷脂的温敏脂质体在肿瘤血管外间质的递送效率及抗肿瘤效果。携cRGD肽靶向的阿霉素温敏脂质体与载IR780的微泡通过生物素亲和素桥连接成复合物,进入循环的复合物通过cRGD肽靶向肿瘤新生血管,富集于肿瘤区域,在局部超声作用下,复合物破碎为小粒径纳米药物,同时肿瘤血管屏障打开,粒径均一的靶向纳米药物可以通过血管内皮间隙进入肿瘤内部,通过cRGD肽与肿瘤细胞上的受体结合,特异性蓄积于肿瘤血管外间质内。同时,肿瘤区域累积的IR780碘化物在激光辐照下引起局部温度升高,达到温敏脂质体释药温度(约42℃)时,阿霉素从脂质体中快速释放,增强化疗药物的生物利用度及抗肿瘤作用。本方法得到的双响应脂质体微泡复合物还具有超声造影成像及荧光成像功能。
附图说明
图1为本方法得到的双响应脂质体微泡复合物的紫外光谱分析图。
图2为本方法得到的双响应脂质体微泡复合物的效果图,其中,A图为所述复合物激光共聚焦的照片,B图为所述复合物的粒径分布曲线图。
图3为本方法得到的双响应脂质体微泡复合物经超声处理后产物的效果图,其中,A图为该产物的透射电镜图,B图为该产物的粒径分布曲线图。
图4为本方法得到的双响应脂质体微泡复合物体外细胞黏附实验及超声靶向成像效果图,其中,A图为光学显微镜下观察非靶向微泡与本方法得到的双响应脂质体微泡复合物(简称复合物)对乳腺癌MCF-7细胞的靶向黏附的显微照片,B图为对MCF-7细胞黏附的微泡数量统计结果的条形图,图中***表示本方法得到的双响应脂质体微泡复合物(简称复合物)与非靶向微泡比,p<0.001;C图为荷瘤裸鼠体内的超声造影声强度统计结果的条形图,图中***表示本方法得到的双响应脂质体微泡复合物(简称复合物)与非靶向微泡比,p<0.001。
图5为本方法得到的双响应脂质体微泡复合物(简称复合物)、生理盐水、携cRGD肽靶向阿霉素温敏脂质体(简称靶向温敏脂质体)和载IR780微泡在荷瘤裸鼠体内抑瘤实验结果的折线图。
图6为本方法得到的双响应脂质体微泡复合物瘤内递送效率的效果图,其中,A图为携cRGD肽靶向阿霉素温敏脂质体(简称靶向温敏脂质体)联合热疗(下图)和本方法得到的双响应脂质体微泡复合物(简称复合物)联合超声及激光辐射(上图)的肿瘤内阿霉素荧光显微照片,B图则为平均荧光强度统计结果的条形图,图中***表示本方法得到的双响应脂质体微泡复合物(简称复合物)与携cRGD肽靶向阿霉素温敏脂质体(简称靶向温敏脂质体)联合热疗比,p<0.001。
具体实施式
实施例1:
、双响应脂质体微泡复合物的制备
(1)称取1×10-5molDSPE-PEG2000-NHS和2×10-5mol的cRGD溶于1mL二甲基甲酰胺(DMF)中,加入0.065mg的三乙胺,室温下搅拌反应4h,真空旋转蒸发去除溶剂及三乙胺后得到DSPE-PEG2000-cRGD。
(2)取1.025×10-4molDPPC、1×10-5molMSPC、1×10-5molDSPE-PEG2000-cRGD和2.5×10-6molDSPE-PEG2000-Biotin溶于1.5mL氯仿中,常温常压旋转混合均匀;真空旋转蒸发去除溶剂,形成一层白色磷脂薄膜,真空干燥,加入pH4.0的柠檬酸盐缓冲液水化处理,待磷脂水化完全后,用孔径为100nm滤膜的脂质体挤出器挤压,得到空载携cRGD肽靶向温敏脂质体纳米粒。
(3)将所得到的靶向温敏脂质体纳米粒的pH值调至7.4,加入600μL阿霉素水溶液(5mg/L)与1.5mL空载靶向温敏脂质体纳米粒溶液(含60mg空载靶向温敏脂质体纳米粒)混匀,37℃水浴20min后,用G-50葡聚糖凝胶柱层析,得到携cRGD肽靶向阿霉素温敏脂质体;其中所述的阿霉素溶液为阿霉素水溶液。
(4)取2mg IR-780加入1mL二甲基亚砜(DMSO)配制成IR780溶液,然后取20μLIR780溶液加入到2mL生物素化脂质微泡中,混匀,4℃条件下400×g离心3min,取上层载IR780脂质微泡;其中,所述的IR780为七甲川花菁染料;
(5)取1mL浓度为109/ml的载IR780脂质微泡和200μL浓度为0.3mg/ml的亲和素混匀,冰上孵育30min,然后加入所述1mL携cRGD肽靶向阿霉素温敏脂质体,轻柔混匀后4℃条件下孵育30min,洗涤,得到所述的双响应脂质体微泡复合物。
、双响应脂质体微泡复合物的鉴定:
将上述步骤(2)纯化后得到的空载携cRGD肽靶向温敏脂质体、步骤(3)得到的携cRGD肽靶向阿霉素温敏脂质体及游离阿霉素进行紫外光谱分析,游离阿霉素在波长480nm处有特征行紫外吸收峰;磷脂的紫外吸收峰在波长220-330nm处;携cRGD肽靶向阿霉素温敏脂质体的紫外光谱图具有阿霉素及磷脂的吸收峰(见图1),说明阿霉素成功装载于携cRGD肽靶向阿霉素温敏脂质体中。
将本发明复合物用去离子水按比例稀释,使用动态光散射仪测量粒径大小;使用激光共聚焦显微镜TRITC通道观察阿霉素荧光的分布位置,确定复合物的形态。激光共聚焦显微镜观察可见复合物粒径大小较均匀,散在分布,无明显聚集现象,TRITC通道激光激发下,阿霉素发射的荧光信号(软件标记为绿色信号)集中出现在微泡外壳层(见图2),表明带携cRGD肽靶向温敏脂质体成功连接于微泡上。本发明复合物经超声辐照处理1分钟后,破碎成形态较均一,平均粒径约60nm的纳米颗粒(见图3)。紫外分光光度法测定载阿霉素温敏脂质体及复合物载药量;温敏脂质体载药量约4.6%,复合物的载药量约20.3μg DOX/108MBs;10.5μg IR780/108MBs。
实施例2:(体内外肿瘤靶向能力检测)
一、样品及对照品
样品为实施例1的双响应脂质体微泡复合物;对照品为非靶向微泡,其制备方法如下,取20μL二甲基亚砜加入到2mL生物素化脂质微泡中,混匀,4℃条件下400×g离心3min,取上层非靶向微泡。
二、体外肿瘤细胞黏附实验
通过比较非靶向微泡与本发明复合物对人乳腺癌MCF-7细胞黏附微泡数量的差异,验证双响应脂质体微泡复合物的肿瘤靶向性。光学显微镜下观察结果发现非靶向微泡组细胞表面没有明显微泡附着现象,而复合物组在细胞的表面可见大量复合物黏附(见图4中A图)。对MCF-7细胞黏附的微泡数量进行统计(见图4中B图),发现复合物组每细胞平均附着微泡数与非靶向微泡组比较具有显著差异(p<0.001)。说明复合物上的cRGD肽配体可以和肿瘤细胞表面的整合素受体结合,且这种结合是特异性且牢固的,复合物具有肿瘤靶向性。
三、荷瘤裸鼠体内超声靶向成像功能检测
通过比较普通非靶向微泡与本复合物的超声造影效果及肿瘤靶向能力,结果显示,爆破前后复合物组肿瘤区域的声强度显著高于非靶向微泡组(见图4中C图),差异具有统计学意义(p<0.001),提示本发明复合物靶向结合于肿瘤组织的微泡量多于非靶向微泡,显示出较强的肿瘤靶向性。
实施例3(体内抑瘤效果检测)
一、样品及对照品
样品为实施例1的双响应脂质体微泡复合物,对照品1为生理盐水;对照品2为实施例1中步骤(1)-(3)得到的携cRGD肽靶向阿霉素温敏脂质体,简称为靶向温敏脂质体;对照品3为实施例1中步骤(4)得到的载IR780微泡。
二、荷瘤动物模型建立
人乳腺癌MCF-7肿瘤组织块无菌条件下解剖分离成2mm×2mm×2mm大小的瘤块,再把瘤组织块移植于裸鼠右侧下背部。
三、实验分组及处理方法
1、实验分组:
(I)生理盐水组;(II)靶向温敏脂质体联合热疗组;(III)载IR780微泡联合超声辐照组;(IV)复合物联合超声及激光辐照组
2、超声辐照方法
药物经尾静脉注射后,在荷瘤裸鼠肿瘤部位涂布适量耦合剂,消除超声治疗仪探头与裸鼠皮肤之间的空气,立即将探头固定于裸鼠肿瘤上,持续超声辐照1分钟。
3、激光辐照方法
药物经尾静脉注射后,使用1%戊巴比妥钠对荷瘤裸鼠进行麻醉,立即将激光探头固定于裸鼠肿瘤部位上方3cm处,持续激光辐照5分钟。
4、温和热疗方法
热疗组在尾静脉注射靶向温敏脂质体后,荷瘤裸鼠给予麻醉处理,将肿瘤部位暴露于43℃温水中进行热疗处理1h,非肿瘤部位使用隔热材料进行保护,热疗过程中注意监测裸鼠体温及心跳呼吸变化。
四、抑瘤实验
各实验组给药量均含等量阿霉素或IR780(按裸鼠体重1.23mg/kg(阿霉素),0.3mg/kg(IR780))或每只裸鼠200μL生理盐水,每5天予以一次相应处理,共治疗三次。治疗过程中,每3天测量一次肿瘤体积。统计及分析各处理组对裸鼠皮下移植瘤的抑制效果。
结果发现复合物联合超声及激光辐照组的抑瘤效果最佳,联合治疗后肿瘤体积缩小,肿瘤生长受到抑制,协同抗肿瘤效应明显优于靶向温敏脂质体联合热疗组及载IR780微泡联合超声辐照组。结果如图5所示。
实施例4(瘤内药物递送效果检测)
一、样品及对照品
样品为实施例1的双响应脂质体微泡复合物,对照品为实施例1步骤(1)-(3)中得到的携cRGD肽靶向阿霉素温敏脂质体,简称为靶向温敏脂质体。
二、荷瘤动物模型建立
荷瘤动物模型建立方法同实施例3。
三、实验分组及处理方法
1、实验分组:
(I)靶向温敏脂质体联合热疗组;(II)复合物联合超声及激光辐照组
2、实验方法
荷瘤裸鼠经尾静脉注射含相同阿霉素剂量的靶向温敏脂质体或复合物(阿霉素/裸鼠体重:3mg/Kg)后,分别进行温和热疗和超声及激光处理(方法同实施例3),药物注射1小时后处死裸鼠并完整分离肿瘤组织,肿瘤组织切片进行DAPI和CD31染色,使用全自动数字切片扫描机检测肿瘤组织内的阿霉素荧光水平。
结果发现复合物联合超声及激光辐照组肿瘤组织内阿霉素平均荧光强度高于靶向温敏脂质体联合热疗组,且复合物组阿霉素多分布于肿瘤中心部位,差异具有统计学意义(p<0.001)。结果如图6所示。
Claims (1)
1.一种载有阿霉素的双响应脂质体微泡复合物的制备方法,该法由以下步骤组成:
(1)按DSPE-PEG2000-NHS︰cRGD=1︰2的摩尔比取DSPE-PEG2000-NHS和cRGD溶于二甲基甲酰胺中,加入DSPE-PEG2000-NHS和cRGD质量和0.2%的催化剂三乙胺,室温下搅拌反应4h,真空旋转蒸发去除溶剂及三乙胺后得到DSPE-PEG2000-cRGD;其中,所述的DSPE-PEG2000-NHS为分子量2000的二棕榈酰磷脂酰乙醇胺-聚乙二醇-琥珀酰亚胺酯,所述的cRGD为环状低聚肽;
(2)按DPPC︰MSPC︰DSPE-PEG2000-cRGD︰DSPE-PEG2000-Biotin=82︰8︰8︰2的摩尔比取DPPC、MSPC、DSPE-PEG2000-cRGD和DSPE-PEG2000-Biotin溶于氯仿中,常温常压旋转混合均匀;然后,室温下抽真空,旋转蒸发挥去溶剂,并在容器底部形成一层白色磷脂薄膜,真空干燥,加入pH4.0的柠檬酸盐缓冲液水化处理,待磷脂水化完全后,用孔径为100nm滤膜的脂质体挤出器挤压,得到空载携cRGD肽的靶向温敏脂质体纳米粒;其中,所述的DPPC为二棕榈酰磷脂酰胆碱,所述的MSPC为溶血磷脂,所述的DSPE-PEG2000-Biotin为生物素化二棕榈酰磷脂酰乙醇胺-聚乙二醇;
(3)将所得到的靶向温敏脂质体纳米粒的pH值调至7.4,然后按阿霉素︰靶向温敏脂质体纳米粒=1︰20的质量比加入阿霉素水溶液,混匀,37℃水浴20min后,用G-50葡聚糖凝胶柱层析,得到携cRGD肽靶向阿霉素温敏脂质体;
(4)取IR-780加入二甲基亚砜配制成浓度为2mg/mL的IR780溶液,然后按IR780溶液︰生物素化脂质微泡=1︰100的体积比将IR780溶液加入到生物素化脂质微泡中,混匀,4℃条件下低速离心,取上层载IR780脂质微泡;其中,所述的IR780为七甲川花菁染料;
(5)按携cRGD肽靶向阿霉素温敏脂质体︰亲和素︰载IR780脂质微泡=5︰1︰5的体积比先取所述载IR780脂质微泡加入亲和素,混匀,冰上孵育30min,然后加入所述携cRGD肽靶向阿霉素温敏脂质体,轻柔混匀后4℃条件下孵育30min,洗涤,得到所述的双响应脂质体微泡复合物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910648607.3A CN110237268B (zh) | 2019-07-18 | 2019-07-18 | 一种载有阿霉素的双响应脂质体微泡复合物的制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910648607.3A CN110237268B (zh) | 2019-07-18 | 2019-07-18 | 一种载有阿霉素的双响应脂质体微泡复合物的制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110237268A true CN110237268A (zh) | 2019-09-17 |
CN110237268B CN110237268B (zh) | 2023-02-03 |
Family
ID=67892688
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910648607.3A Active CN110237268B (zh) | 2019-07-18 | 2019-07-18 | 一种载有阿霉素的双响应脂质体微泡复合物的制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110237268B (zh) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110859971A (zh) * | 2019-10-21 | 2020-03-06 | 哈尔滨医科大学 | 一种携ir-780的靶向声释氧纳米微聚体及其制备方法和应用 |
CN111249471A (zh) * | 2020-01-18 | 2020-06-09 | 广州医科大学附属第三医院(广州重症孕产妇救治中心、广州柔济医院) | 一种基因递送的聚乙烯亚胺纳米粒微泡复合物的制备方法 |
CN113144172A (zh) * | 2021-02-22 | 2021-07-23 | 温州医科大学附属第二医院(温州医科大学附属育英儿童医院) | 一种含有万古霉素、ir780与携氧全氟己烷的脂质体的制备方法 |
CN114632071A (zh) * | 2022-03-23 | 2022-06-17 | 长春工业大学 | 一种脂质体纳米粒子及其制备方法和应用 |
CN115006368A (zh) * | 2022-07-01 | 2022-09-06 | 重庆大学 | 细胞膜包被纳米药物及其应用 |
CN115040526A (zh) * | 2022-06-23 | 2022-09-13 | 重庆大学 | 一种靶向纳米药物复合物及其制备方法 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020102298A1 (en) * | 1998-06-18 | 2002-08-01 | David Needham | Temperature-sensitive liposomal formulation |
CN101327190A (zh) * | 2008-07-29 | 2008-12-24 | 北京大学 | 一种供注射用的抗肿瘤长循环靶向脂质体 |
CN101485629A (zh) * | 2008-01-16 | 2009-07-22 | 沈阳药科大学 | 一种给药系统及其制备方法 |
CN101864071A (zh) * | 2010-05-21 | 2010-10-20 | 北京中海康医药科技发展有限公司 | 聚乙二醇-二硬脂酰磷脂酰乙醇胺衍生物及其制备方法 |
CN106166299A (zh) * | 2016-09-14 | 2016-11-30 | 南方医科大学 | 一种载有阿霉素的可视化微泡复合物及其制备方法 |
CN107847444A (zh) * | 2015-05-26 | 2018-03-27 | 通用医疗公司 | 脂质体纳米构建体及其制备和使用方法 |
-
2019
- 2019-07-18 CN CN201910648607.3A patent/CN110237268B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020102298A1 (en) * | 1998-06-18 | 2002-08-01 | David Needham | Temperature-sensitive liposomal formulation |
CN101485629A (zh) * | 2008-01-16 | 2009-07-22 | 沈阳药科大学 | 一种给药系统及其制备方法 |
CN101327190A (zh) * | 2008-07-29 | 2008-12-24 | 北京大学 | 一种供注射用的抗肿瘤长循环靶向脂质体 |
CN101864071A (zh) * | 2010-05-21 | 2010-10-20 | 北京中海康医药科技发展有限公司 | 聚乙二醇-二硬脂酰磷脂酰乙醇胺衍生物及其制备方法 |
CN107847444A (zh) * | 2015-05-26 | 2018-03-27 | 通用医疗公司 | 脂质体纳米构建体及其制备和使用方法 |
CN106166299A (zh) * | 2016-09-14 | 2016-11-30 | 南方医科大学 | 一种载有阿霉素的可视化微泡复合物及其制备方法 |
Non-Patent Citations (6)
Title |
---|
WANXIAN LUO,ET AL: ""Dual-targeted and pH-sensitive Doxorubicin Prodrug-Microbubble Complex with Ultrasound for Tumor Treatment "", 《THERANOSTICS》 * |
孙飞等: "热敏脂质体的研究进展", 《药学进展》 * |
杨阳等: ""优化制备生物素化超声微泡的实验研究"", 《昆明医科大学学报》 * |
王冬晓等: ""超声辐照联合双配体载药纳米粒对人乳腺癌耐紫杉醇细胞株细胞毒性效果评价"", 《临床超声医学杂志》 * |
罗婉贤等: ""超声辐照联合酸敏双配体载阿霉素前药-微泡复合物体内抗肿瘤特性研究"", 《临床超声医学杂志》 * |
陆媛媛等: "热敏脂质体在肿瘤靶向治疗中的研究进展", 《中国医院药学杂志》 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110859971A (zh) * | 2019-10-21 | 2020-03-06 | 哈尔滨医科大学 | 一种携ir-780的靶向声释氧纳米微聚体及其制备方法和应用 |
CN111249471A (zh) * | 2020-01-18 | 2020-06-09 | 广州医科大学附属第三医院(广州重症孕产妇救治中心、广州柔济医院) | 一种基因递送的聚乙烯亚胺纳米粒微泡复合物的制备方法 |
CN113144172A (zh) * | 2021-02-22 | 2021-07-23 | 温州医科大学附属第二医院(温州医科大学附属育英儿童医院) | 一种含有万古霉素、ir780与携氧全氟己烷的脂质体的制备方法 |
CN113144172B (zh) * | 2021-02-22 | 2023-10-03 | 温州医科大学附属第二医院(温州医科大学附属育英儿童医院) | 一种含有万古霉素、ir780与携氧全氟己烷的脂质体的制备方法 |
CN114632071A (zh) * | 2022-03-23 | 2022-06-17 | 长春工业大学 | 一种脂质体纳米粒子及其制备方法和应用 |
CN115040526A (zh) * | 2022-06-23 | 2022-09-13 | 重庆大学 | 一种靶向纳米药物复合物及其制备方法 |
CN115006368A (zh) * | 2022-07-01 | 2022-09-06 | 重庆大学 | 细胞膜包被纳米药物及其应用 |
CN115006368B (zh) * | 2022-07-01 | 2023-03-14 | 重庆大学 | 细胞膜包被纳米药物及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN110237268B (zh) | 2023-02-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110237268A (zh) | 一种载有阿霉素的双响应脂质体微泡复合物的制备方法 | |
Song et al. | Liposomes co-loaded with metformin and chlorin e6 modulate tumor hypoxia during enhanced photodynamic therapy | |
Son et al. | Folate-modified PLGA nanoparticles for tumor-targeted delivery of pheophorbide a in vivo | |
Wahab et al. | Current trends and future perspectives of nanomedicine for the management of colon cancer | |
CN108354901A (zh) | 用于肿瘤化疗与光热联合治疗的pH/还原双重敏感多功能纳米胶束及其应用 | |
US9844656B2 (en) | Localization of agents at a target site with a composition and an energy source | |
CN108653733B (zh) | 具有气泡生成功能的双载蒽环类药物及光敏剂的聚合物囊泡与制备 | |
Yang et al. | NIR-activated self-sensitized polymeric micelles for enhanced cancer chemo-photothermal therapy | |
Rao et al. | Size-adjustable micelles co-loaded with a chemotherapeutic agent and an autophagy inhibitor for enhancing cancer treatment via increased tumor retention | |
CN109771391A (zh) | 血小板膜包被的阿霉素-吲哚菁绿仿生纳米颗粒及其用途 | |
CN108578711B (zh) | 一种乙酰化糖酯-聚乙二醇-磷脂酰乙醇胺共轭物及其制备方法与应用 | |
CN108653754A (zh) | 一种透明质酸靶向聚多巴胺包覆相变型液态氟碳纳米超声造影剂 | |
CN107050040A (zh) | Hifu控释的脑胶质瘤靶向纳米递药系统及其制备方法和用途 | |
CN107019801A (zh) | 一种磁热释放的热敏脂质体 | |
CN108938594A (zh) | 一种药物复合物及其制备方法与应用 | |
Li et al. | iRGD peptide-mediated liposomal nanoparticles with photoacoustic/ultrasound dual-modality imaging for precision theranostics against hepatocellular carcinoma | |
CN112535676A (zh) | 提高阿霉素肿瘤主动靶向性和肾脏保护的纳米结构脂质制剂及制备方法 | |
Liu et al. | Liposome-based multifunctional nanoplatform as effective therapeutics for the treatment of retinoblastoma | |
CN113440610B (zh) | 共载cd73抗体和阿霉素的脂质体及其制备方法和应用 | |
Su et al. | Photo-responsive NIR-II biomimetic nanomedicine for efficient cancer-targeted theranostics | |
CN110448699A (zh) | 包含功能性多肽修饰七甲川花菁素类染料的肿瘤细胞核靶向载药纳米粒子及制备方法 | |
CN104546722B (zh) | 米铂脂质体和制法 | |
CN109675052B (zh) | 生物点击触发的高效靶向偶联物及其多元组合物、制备方法和应用 | |
Wang et al. | Dual-functional melanin-based nanoliposomes for combined chemotherapy and photothermal therapy of pancreatic cancer | |
WO2022228230A1 (zh) | 一种双亲性材料及其在制备脂质体中的应用 |
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 |