CN114099709B - 一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 - Google Patents
一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 Download PDFInfo
- Publication number
- CN114099709B CN114099709B CN202111475996.8A CN202111475996A CN114099709B CN 114099709 B CN114099709 B CN 114099709B CN 202111475996 A CN202111475996 A CN 202111475996A CN 114099709 B CN114099709 B CN 114099709B
- Authority
- CN
- China
- Prior art keywords
- porphyrin
- doxorubicin
- selenoether
- organic polymer
- composite nano
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229960004679 doxorubicin Drugs 0.000 title claims abstract description 113
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 239000002105 nanoparticle Substances 0.000 title claims description 79
- 239000002131 composite material Substances 0.000 title claims description 72
- 229920000620 organic polymer Polymers 0.000 claims abstract description 48
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 45
- 238000001027 hydrothermal synthesis Methods 0.000 claims abstract description 10
- 150000004032 porphyrins Chemical class 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims description 61
- 229940079593 drug Drugs 0.000 claims description 42
- 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 claims description 36
- 229960002918 doxorubicin hydrochloride Drugs 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 19
- UVAJHKGWZQPAEZ-UHFFFAOYSA-N C12=CC=C(N1)C=C1C=CC(=N1)C=C1C=CC(N1)=CC=1C=CC(N1)=C2.NC2=CC=CC=C2 Chemical compound C12=CC=C(N1)C=C1C=CC(=N1)C=C1C=CC(N1)=CC=1C=CC(N1)=C2.NC2=CC=CC=C2 UVAJHKGWZQPAEZ-UHFFFAOYSA-N 0.000 claims description 13
- XIMIGUBYDJDCKI-UHFFFAOYSA-N diselenium Chemical compound [Se]=[Se] XIMIGUBYDJDCKI-UHFFFAOYSA-N 0.000 claims description 10
- 239000004815 dispersion polymer Substances 0.000 claims description 10
- 239000007864 aqueous solution Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 8
- 239000011248 coating agent Substances 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- 230000000379 polymerizing effect Effects 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 3
- 206010060862 Prostate cancer Diseases 0.000 claims description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 3
- 230000000973 chemotherapeutic effect Effects 0.000 claims description 2
- 239000003596 drug target Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000001291 vacuum drying Methods 0.000 claims description 2
- 230000001659 chemokinetic effect Effects 0.000 claims 1
- 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 abstract description 75
- 238000011282 treatment Methods 0.000 abstract description 18
- 230000008685 targeting Effects 0.000 abstract description 13
- 239000000463 material Substances 0.000 abstract description 12
- 229920000642 polymer Polymers 0.000 abstract description 9
- 239000000126 substance Substances 0.000 abstract description 7
- 239000002246 antineoplastic agent Substances 0.000 abstract description 5
- 210000001519 tissue Anatomy 0.000 abstract description 5
- 229940044683 chemotherapy drug Drugs 0.000 abstract description 4
- 230000002035 prolonged effect Effects 0.000 abstract description 4
- 230000002829 reductive effect Effects 0.000 abstract description 4
- 230000017531 blood circulation Effects 0.000 abstract description 3
- 239000003642 reactive oxygen metabolite Substances 0.000 abstract 2
- 210000000805 cytoplasm Anatomy 0.000 abstract 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 22
- 230000000694 effects Effects 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 18
- 229960003180 glutathione Drugs 0.000 description 11
- 238000011068 loading method Methods 0.000 description 10
- 239000000872 buffer Substances 0.000 description 9
- 201000011510 cancer Diseases 0.000 description 6
- 238000005538 encapsulation Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 238000011580 nude mouse model Methods 0.000 description 6
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 6
- 241000699660 Mus musculus Species 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000002329 infrared spectrum Methods 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 229940124597 therapeutic agent Drugs 0.000 description 5
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 150000002466 imines Chemical class 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 3
- 229940009456 adriamycin Drugs 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000001186 cumulative effect Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000013558 reference substance Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 206010067484 Adverse reaction Diseases 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 206010048610 Cardiotoxicity Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000003917 TEM image Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 231100000259 cardiotoxicity Toxicity 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 230000009036 growth inhibition Effects 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 238000001931 thermography Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 201000004384 Alopecia Diseases 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 206010065553 Bone marrow failure Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000031229 Cardiomyopathies Diseases 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 208000012641 Pigmentation disease Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- HPZOOQSXPMEJBV-ODCFVKFUSA-N Tirilazad mesylate Chemical compound CS(O)(=O)=O.O=C([C@@H]1[C@@]2(C)CC=C3[C@@]4(C)C=CC(=O)C=C4CC[C@H]3[C@@H]2C[C@H]1C)CN(CC1)CCN1C(N=1)=CC(N2CCCC2)=NC=1N1CCCC1 HPZOOQSXPMEJBV-ODCFVKFUSA-N 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 231100000360 alopecia Toxicity 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000003972 antineoplastic antibiotic Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000011284 combination treatment Methods 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 208000001297 phlebitis Diseases 0.000 description 1
- 239000003504 photosensitizing agent Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 208000003265 stomatitis Diseases 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000001757 thermogravimetry curve Methods 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 230000008673 vomiting 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
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nanotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Manufacturing & Machinery (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Composite Materials (AREA)
- Materials Engineering (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种用于化学‑光热‑化学动力学联合肿瘤治疗的光热纳米制剂的制备方法及其应用,该方法首先在通过水热反应得到卟啉多孔共价有机聚合物,将阿霉素封装在其中,制备获得光敏性材料,通过利用多孔共价聚合物存在的特异性环境敏感的共价键被动靶向,不仅可以有效延长其血液循环时间,而且可以在肿瘤部位有效富集。到达肿瘤组织后,在激光的照射下,多孔有机聚合物以吸收激光的能量并转化为热能,随后光敏性多孔有机聚合物会加速释放化疗药物阿霉素,同时双硒键被肿瘤细胞质过表达的GSH还原产生活性氧(ROS),从而实现光热治疗‑化疗‑化学动力学的联合肿瘤治疗。
Description
技术领域
本发明涉及生物制药技术领域,尤其涉及一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用。
背景技术
靶向药物(也称作靶向制剂)是指被赋予了靶向(Targeting)能力的药物或其制剂。其目的是使药物或其载体能瞄准特定的病变部位,并在目标部位蓄积或释放有效成分。靶向制剂可以使药物在目标局部形成相对较高的浓度,从而在提高药效的同时抑制毒副作用,减少对正常组织、细胞的伤害。
近红外光热治疗剂是指利用在近红外光区具有较高光热转换效率的试剂,将其通过血管注射入生物内部,利用靶向性识别技术聚集在肿瘤组织附近,并在近红外光的照射下将光能转化为热能来杀死癌细胞的一种治疗试剂。但是目前的光热治疗剂的靶向性差近红外区吸收能力差等特点,限制了光热治疗剂在临床中的应用。
阿霉素(DOX)是在恶性肿瘤临床治疗中被广泛使用的小分子化学治疗药物,作为一种蒽环类抗肿瘤抗生素,阿霉素具有较广的抗瘤谱和较强的抗肿瘤疗效,但是有许多研究表明,阿霉素具有非常严重的不良反应和副反应,如心脏毒性、骨髓抑制、恶心、呕吐、口腔炎和脱发等。晚期的心脏毒性则与其用药剂量相关,是不可逆转的严重的心肌病变。60%~80%的恶性肿瘤患者注射阿霉素之后可能会发生骨髓抑制,临床表现为白细胞相应减少、血小板一定水平的减少甚至贫血。除此之外,DOX的不良反应还有静脉炎、皮肤色素沉着、肝脏功能的损害等,临床应用受到很大限制。而且由于药物本身的非靶向性,会无差异的在人体分布,利用率低,达不到理想的治疗效果。
因此,如何提供一种具有优异有效性和安全性的制剂材料,在增加药物被动靶向效果,实现药物在肿瘤内部的特异性释放,增强药物的释放,提高药物的利用度,提高治疗效果的同时,降低药物的毒副作用,对癌症治疗意义重大。
发明内容
本发明的目的在于提供一种光热和化学联合肿瘤治疗的响应性释放药物光热制剂,即所述卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用。通过利用多孔共价聚合物存在得特异性环境敏感的共价键被动靶向,不仅可以有效延长其血液循环时间,而且可以在肿瘤部位有效富集。到达肿瘤组织后,在激光的照射下,多孔有机聚合物以吸收激光的能量并转化为热能,随后热敏性多孔有机聚合物会加速释放化疗药物阿霉素,从而实现光热治疗和化疗的联合肿瘤治疗。
本发明的第一个目的是提供一种卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,所述方法包括将5,10,15,20-四(4-氨基苯)-21H,23H-卟啉和二硒醚聚合,形成卟啉-硒醚多孔有机聚合物,将盐酸阿霉素包覆在所述卟啉多孔有机聚合物内。
进一步的,所述方法包括以下步骤:
1)将盐酸阿霉素溶解于水中,得到盐酸阿霉素的水溶液;
2)将5,10,15,20-四(4-氨基苯)-21H,23H-卟啉与二硒醚通过水热反应进行聚合,真空干燥,得到卟啉-硒醚多孔有机聚合物;
将所述卟啉-硒醚多孔有机聚合物分散于水中,得到卟啉-硒醚多孔有机聚合物分散溶液;
3)将步骤2)所述卟啉-硒醚多孔有机聚合物分散溶液与所述步骤1)中盐酸阿霉素溶液混合,剧烈搅拌直至完成包覆,得到卟啉-硒醚-阿霉素复合纳米颗粒。
所述步骤1)和2)没有先后顺序限制。
进一步的,步骤1)所述盐酸阿霉素的水溶液的浓度为1~10mg/mL,优选的,所述盐酸阿霉素的水溶液的浓度为1mg/mL。
进一步的,步骤2)所述5,10,15,20-四(4-氨基苯)-21H,23H-卟啉与二硒醚的物质的摩尔比为1:2。
进一步的,步骤2)所水热反应的溶剂为1,4二氧六环,所述溶剂用量与5,10,15,20-四(4-氨基苯)-21H,23H-卟啉摩尔比为700:1,水热反应的条件为100℃~160℃,12~36h,优选的,水热反应条件为120℃,24h。
进一步的,步骤2)所述卟啉-硒醚多孔有机聚合物分散溶液浓度为1mg/mL。
进一步的,步骤3)所述卟啉-硒醚多孔有机聚合物分散溶液与盐酸阿霉素溶液的质量比为1:0.25~1:5,优选的,所述卟啉-硒醚多孔有机聚合物分散溶液与盐酸阿霉素溶液的质量比为1:2。
进一步的,步骤3)所述搅拌时间为8~24h,优选的,所述搅拌时间为12h;所述搅拌在黑暗环境下进行。
本发明的第二个目的是提供基于前述卟啉-硒醚-阿霉素复合纳米颗粒的制备方法得到的卟啉-硒醚-阿霉素复合纳米颗粒。
本方法采用全新的材料来包覆阿霉素,不同于线性聚合物,卟啉-硒醚多孔有机聚合物具有固有的多孔结构,可以实现对药物的高效吸附,大大提高载药量,解决线性聚合物载药量低的难题。相比于小分子的卟啉光敏剂,聚合物具有更大的共轭结构,光热性能更强,更好的发挥药物跟载体的协同作用。此外,亚胺跟二硒的同时引入,使得材料对癌症微环境的响应性更强。卟啉单元、动态双Se键、亚胺键的同时引入使材料具有光-GSH-pH三重响应性,几乎可以实现载入药物的完全释放,达到光-化学-化学动力学疗法协同高效治疗肿瘤的目的。本发明的第三个目的是提供前述的卟啉-硒醚-阿霉素复合纳米颗粒在制备靶向肿瘤药物中的应用。
优选的,所述靶向肿瘤药物为同时具有光热治疗、化学治疗、化学动力学治疗效果的药物;
优选的,所述靶向肿瘤药物靶向治疗前列腺癌的肿瘤药物。
所述卟啉-硒醚-阿霉素复合纳米颗粒中的动态共价键二硒键的氧化还原响应靶向释放阿霉素,从而可以实现氧化还原敏感的释药。动态亚胺键,可实现的pH响应性释药。所述卟啉-硒醚-阿霉素复合纳米颗粒还可以发挥良好的光热性能,通过近红外光照射可以转换为热能,促进药物释放。从而实现光-GSH-pH三重响应性的释放,药物释放量可以接近85%。
本发明采用上述技术方案,与现有技术相比,具有如下技术效果:
本发明为了提高抗肿瘤药物的靶向性和在体内的肿瘤抑制效率,设计了一种封装阿霉素的卟啉-硒醚-阿霉素复合纳米颗粒,将具有特异性质的单体聚合制备同时具有亚胺和双硒动态共价键的多孔聚合物,得到具有光、GSH和pH三重响应性的智能载体。利用多孔结构实现对药物的有效负载,实现光热-化学-化学动力学联合治疗癌症。载体与盐酸阿霉素结合后,可以得到更优越的光热性能,升温后会进一步增强药物的释放。通过材料的包覆,降低药物的毒副作用,增加其被动靶向,实现药物在肿瘤内部的特异性释放,提高阿霉素的利用度,提高治疗效果意义重大。另外,双Se键的存在可以实现优越的化学动力疗法,进而实现三位一体治疗,药物释放量可以接近85%。
本发明提供卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,首先在通过水热反应得到卟啉-硒醚多孔有机聚合物,将阿霉素封装在其中,制备获得光敏性材料,通过利用多孔共价聚合物存在得特异性环境敏感的共价键被动靶向,相比于单独的盐酸阿霉素,纳米颗粒不仅可以有效延长其血液循环时间,而且可以在肿瘤部位有效富集。到达肿瘤组织后,在激光的照射下,多孔有机聚合物可以吸收激光的能量并转化为热能,体内外光热性能优越,808nm激光照射5min可以达到54.4℃;体内照射5min可达到52.3℃。随后光敏性多孔有机聚合物会加速释放化疗药物阿霉素,从而实现光热治疗-化疗-化学动力学的联合肿瘤治疗。
本发明制备的封装阿霉素的卟啉-硒醚-阿霉素复合纳米颗粒具有毒性低、绿色环保、研制成本低、可生物相容等优点,是一种制备新型光热治疗剂的理想材料,同时还可以用作化疗药物载体。聚合形成的共价键更有利于肿瘤细胞对高还原性谷胱甘肽(GSH)表达、低pH等特定肿瘤微环境的响应,实现被动靶向的药物释放。
附图说明
图1为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒的电镜图。
图2为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒的红外吸收光谱图。
其中a)为单体及卟啉-硒醚多孔有机聚合物的红外图谱;b)为卟啉-硒醚多孔有机聚合物以及卟啉-硒醚-阿霉素复合纳米颗粒的红外图谱。
图3为实施例3(2)得到的盐酸阿霉素的药物标准曲线。
图4为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒对不同pH,还原环境下的响应性释药曲线。
其中a)在不同pH值的缓释缓冲液中卟啉-硒醚-阿霉素复合纳米颗粒中盐酸阿霉素的累积释放曲线。(b)在不同浓度GSH的释放缓冲液(pH7.4)中,卟啉-硒醚-阿霉素复合纳米颗粒中盐酸阿霉素的累积释放曲线。(c)在不同浓度GSH的释放缓冲液(pH5.5)中,卟啉-硒醚-阿霉素复合纳米颗粒中盐酸阿霉素的累积释放曲线。(d)在释放缓冲液中,卟啉-硒醚-阿霉素复合纳米颗粒加激光照射干预对DOX的累积释放曲线。
图5为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒在808nm激光照射6分钟的条件下,卟啉-硒醚多孔有机聚合物和卟啉-硒醚-阿霉素复合纳米颗粒升温曲线图。
其中a)为不同浓度卟啉-硒醚多孔有机聚合物在1.5W/cm2功率下照射6分钟温度记录图;b)为不同浓度卟啉-硒醚-阿霉素复合纳米颗粒在1.5W/cm2功率下照射6分钟温度记录图。
图6为本实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒在808nm激光照射5分钟的条件下的动物体内红外热成像图。
图7为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒细胞毒性试验
其中a)游离DOX对PC-3细胞存活率的影响的示意图;b)卟啉-硒醚-阿霉素复合纳米颗粒对PC-3细胞存活率的影响的示意图;c)卟啉-硒醚-阿霉素复合纳米颗粒在1.5W/cm2光照5分钟条件下对PC-3细胞存活率的影响的示意图。
图8为卟啉-硒醚-阿霉素复合纳米颗粒与盐酸阿霉素的药物代谢动力学曲线。
图9为实施例1制备得到的卟啉-硒醚-阿霉素复合纳米颗粒肿瘤抑制试验
其中a)肿瘤治疗期各组肿瘤体积的变化的示意图,肿瘤体积每隔一天测量一次;b)肿瘤治疗期间裸鼠的体重变化的示意图。
图10为光热-化学-化学动力联合肿瘤治疗的示意流程图
具体实施方式
下面结合附图和实施例,对本发明的具体实施方式作进一步描述。以下实施例仅用于更加清楚地说明本发明的技术方案,而不能以此来限制本发明的保护范围。
实施例1制备卟啉-硒醚-阿霉素复合纳米颗粒
步骤a)将盐酸阿霉素溶解于水中,得到1mg/mL的盐酸阿霉素的水溶液;
步骤b)称量5,10,15,20-四(4-氨基苯)-21H,23H-卟啉(0.338g,0.5mmol),购于江苏艾康生物医药研发有限公司、二硒醚(0.368g,1mmol)、于30mL的1,4-二氧六环溶液中溶解,放入高温玻璃管中,超声反应30min,经三次冷冻泵-解冻循环后空密封。反应管在120℃恒温反应24h后,完成水热反应。冷却并打开,过滤分离紫色产物。进一步纯化时,用四氢呋喃(THF)洗脱紫色未反应产物,并进一步在真空干燥过后得到卟啉-硒醚多孔有机聚合物;
步骤c)将步骤b)得到的到卟啉-硒醚多孔有机聚合物分散在水中,对水化溶液进行超声处理以形成均匀的卟啉-硒醚多孔有机聚合物悬浮液,浓度为1mg/mL;
步骤d)将质量比为2:1的步骤a)所获得的盐酸阿霉素溶液与步骤c)获得的卟啉-硒醚多孔有机聚合物悬浮液混合,在冰浴中,用500w的功率超声2h(超声波频率:19-25kHz,3s的时间间隔),使得载体与阿霉素充分混合。然后在黑暗环境下剧烈搅拌12小时,离心收集卟啉-硒醚-阿霉素纳米颗粒(11000rpm,10分钟),得到水分散体系。然后,在黑暗中,透析除去多余未包载的阿霉素。冻干后得到了制备的纳米颗粒。
实施例2卟啉-硒醚多孔有机聚合物以及卟啉-硒醚-阿霉素复合纳米颗粒形态学特征及表征研究
将实施例1步骤b)制备得到卟啉-硒醚多孔有机聚合物以及实施例1步骤d)制备得到卟啉-硒醚-阿霉素纳米颗粒进行形态学研究。室温下透射电镜(TEM)观察其形态(图1)。
采用溴化钾压片法检测样品的红外吸收光谱。将卟啉-硒醚-阿霉素聚合物,阿霉素,卟啉-硒醚-阿霉素纳米颗粒固体粉末分别与溴化钾粉未混合研磨,取1-2mg样品加100-200mg溴化钾在玛瑙硏钵中硏成细粉,研磨时不断用小不锈钢铲把样品刮至研钵中心,以便硏磨得更细避兔颗粒不均匀产生散射造成基线不平。硏磨5-15min即可压片,油压机压力通常为8000-15000kg/cm2加压时间至少保持1min,得到透明锭片。然后将锭片至于傅里叶红外光谱仪之中,检测其吸收波长。(图2)
图1上排由左至右分别为卟啉-硒醚多孔有机聚合物(DiSe-Por)在1μm、200nm、100nm、5nm比例尺下的TEM图,下排由左至右分别为卟啉-硒醚-阿霉素复合纳米颗粒(DiSe-Por-DOX)在1μm、200nm、100nm、5nm比例尺下的TEM图。
图2为卟啉-硒醚多孔有机聚合物和卟啉-硒醚-阿霉素复合纳米颗粒的红外光谱。
结果显示:卟啉-硒醚多孔有机聚合物具有多孔性状,加载阿霉素之后仍保持多孔性状(图1)。卟啉-硒醚多孔有机聚合物的红外光谱综合了5,10,15,20-四(4-氨基苯)-21H,23H-卟啉和二硒醚的特征,其中,二硒醚在3425cm-1(C=O),1635cm-1(C=C),1134cm-1,833cm-1(Se-Se)和584cm-1(Se-C)处的特征峰,以及5,10,15,20-四(4-氨基苯)-21H,23H-卟啉中C=N(1600-1)和N-H(3378cm-1)的特征峰均可在卟啉-硒醚多孔有机聚合物中观察到。除此之外,,10,15,20-四(4-氨基苯)-21H,23H-卟啉的氨基(3378cm-1)和二硒醚中醛(3425cm-1)的吸收峰几乎消失,取而代之的是亚胺(1610cm-1)的吸收峰,这归因于胺和醛的聚合。除了上述提到的卟啉-硒醚多孔有机聚合物的特征峰外,卟啉-硒醚-阿霉素复合纳米颗粒的红外光谱中还可以观察到盐酸阿霉素在2930,1623cm-1的特征吸收峰(图2)。
此外,实施例1制备的卟啉-硒醚-阿霉素复合纳米颗粒粒径范围为~200nm~400nm。
实施例3实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒包封率、载药量的测定
(1)称取阿霉素对照品适量,蒸馏水溶解后紫外分光光度仪在200~800nm进行扫描,得最大吸收波长在483nm处。
(2)标准曲线的绘制
用蒸馏水配制0.5g/L的盐酸阿霉素标准溶液储备液。分别取得质量浓度为5.0、10.0、20.0、30.0、40.0、50.0μg/L的标准溶液供试液。在483nm波长处分别测定不同质量浓度盐酸阿霉素供试液的吸光度(A)值,以A值对质量浓度进行线性回归,回归方程为A=0.0179C+0.0167(R2=0.9998,n=3),线性关系良好。
(3)卟啉-硒醚-阿霉素复合纳米颗粒的包封率、载药量的测定
将制实施例1得到的步骤d)“透析除去多余未包载的阿霉素”前的水分散体系,在10000r/min超速离心20min。收集上清,通过紫外可见光光度计测定483nm下的吸光度(A)值,带入标准曲线计算游离阿霉素的量。阿霉素包封率与载药量分别按下式计算,结果包封率和载药量分别为(35.12±2.251)%和(27.15±2.65)%(n=3)。
包封率=(阿霉素总量-游离阿霉素的量)/阿霉素总量
载药量=(阿霉素总量-游离阿霉素的量)/纳米粒子的量
结果显示:紫外检测到的盐酸阿霉素吸光度,计算得出的标准曲线为y=0.0179x+0.0167(R2=0.9998)。卟啉-硒醚多孔有机聚合物与盐酸阿霉素聚合物质量比例为1:2时,计算得出包封率和载药量分别为(35.12±2.251)%和(27.15±2.65)%(n=3)。(图3)
实施例4实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒中阿霉素的响应性体外释放
为研究实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒在不同介质的影响下的释放行为,取卟啉-硒醚-阿霉素纳米制剂悬液于透析袋中,采用透析法测盐酸阿霉素的体外释放。转速50r/min,温度(37.0±0.5)℃。
释放介质:
a)PBS缓冲液(pH 7.4)
b)PBS缓冲液(pH 6.5)
c)PBS缓冲液(pH 5.5)
d)PBS缓冲溶液(pH7.4+GSH)
e)PBS缓冲溶液(pH5.5+GSH)
f)PBS缓冲溶液(pH5.5+GSH+808nm激光照射)。
间歇收集释放液1mL,同时补充等量的释放介质。将收集的释放液稀释,其中阿霉素的定量测定方法见实施例3中的(1)(2)。卟啉-硒醚-阿霉素复合纳米制剂的体外释放(n=3)结果见图3。
结果显示,可以看出,实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒在pH=5.5和pH=6.5的酸性环境下可以实现更多的药物释放(图4a)。释药行为也与GSH的浓度呈正相关,GSH浓度越高,释放药物越多,证明可以更好的响应肿瘤环境(图4b)在更近似肿瘤微环境的酸性(pH=5.5)和高GSH表达(GSH=10mM)(图4c)的环境下,药物释放更完全,可以达到更好的靶向释药结果。可以看出纳米制剂具有较好的缓释效果,符合水不溶性骨架的释药性能,适合应用于缓控释药物制剂。并且在特定的模拟肿瘤微环境中,高效释放药物。施加激光照射干预可以进一步增强药物释放(图4d)。
实施例5光热性能研究
对实施例1步骤b)中制备的卟啉-硒醚多孔有机聚合物,实施例1步骤d)中制备的卟啉-硒醚-阿霉素复合纳米颗粒的相关光热性能进行如下研究:
将500μL不同浓度(0、100μg/ml、200μg/ml、400μg/ml、600μg/ml)卟啉-硒醚多孔有机聚合物水溶液和卟啉-硒醚-阿霉素复合纳米颗粒水溶液加入管中,用808nm的(1.5W/cm2)激光照射6min。
使用热成像仪(FLIRE5,FLIRSystemAB,Sweden)记录温度变化。记录了材料在辐照和激光关闭下的温度变化,0~5分钟动物体内红外热成像结果如图6所示。然后根据之前的报告计算光热转换效率,结果见图5。
结果显示:可以看得出卟啉-硒醚多孔有机聚合物和卟啉-硒醚-阿霉素复合纳米颗粒在1.5W/cm2功率下照射6分钟,在浓度为600μg/ml时,加载阿霉素之后的卟啉-硒醚-阿霉素复合纳米颗粒升温更明显,显现出了更优异的光热转换性能。
实施例6MTT法测定实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒作为靶向纳米制剂对PC-3细胞的抑制率
(1)细胞培养
人前列腺癌PC-3细胞株(来源:ATCC)培养于含10%胎牛血清的RPMI 1640培养基中,置于5%CO2细胞培养箱中,恒温37℃培养。每1~2d换液1次,使用0.25%胰蛋白酶消化传代,取对数生长期的细胞进行实验。
(2)MTT法测定实施例1制得的卟啉-硒醚-阿霉素复合纳米颗粒对PC-3细胞的抑制率
取(1)中处于对数生长期的PC-3,胰酶消化后进行细胞计数并将细胞的浓度调整为1×105个/mL,用新鲜培养液制备成单细胞悬液,按照每孔100μL接种于96孔板中,放置于37℃、含5%CO2的二氧化碳培养箱中培养24h观察细胞贴壁情况,弃去原培养液,加入不同浓度的卟啉-硒醚-阿霉素复合纳米颗粒(其中所含阿霉素浓度分别为0.125,0.25,0.5,1,2,3,4,5μg/mL)。共同培养12小时后,用1.5W/cm2的808mm激光器照射5分钟,然后,将细胞再孵育24小时进行MTT测定以测量细胞活力,抑制作用结果见图7。
细胞增殖抑制率=(对照组平均A值-给药组平均A值)/对照组平均A值
由结果可知,各组盐酸阿霉素制剂在不同时间作用于PC-3细胞后,细胞的生长均受到不同程度明显的抑制作用,随着制剂浓度增大,作用时间延长,细胞的生长抑制作用也更为明显,可以看出卟啉-硒醚-阿霉素符合纳米制剂对PC-3细胞的生长抑制作用具有较明显的量效和时效关系;施加808nm激光照射后,可以发挥更好的光热作用,达到杀伤细胞的作用。
实施例7卟啉-硒醚-阿霉素复合纳米颗粒的药物代谢动力学研究
将卟啉-硒醚-阿霉素复合纳米颗粒与盐酸阿霉素(其中盐酸阿霉素的浓度均为2.5mg/kg)尾静脉注入大鼠体内,在不同时间尾静脉取大鼠血液,检测血液中盐酸阿霉素的含量。详细结果见图8。
由结果可知,注射卟啉-硒醚-阿霉素复合纳米颗粒后,药物在大鼠体内的循环时间远远优于单纯的盐酸阿霉素。这可以延长药物在体内作用的时间,可以更多的将药物积累在肿瘤部位,达到更好的治疗效果。
实施例8卟啉-硒醚-阿霉素复合纳米颗粒体内联合治疗性能评估:
将100uL(200万)的PC-3细胞皮下接种于3-5周的雄性裸鼠右侧大腿处,带肿瘤体积长到100mm3时,将裸鼠分为8组(每组5只):(1)生理盐水、(2)游离的阿霉素、(3)卟啉-硒醚多孔有机聚合物、(4)卟啉-硒醚-阿霉素复合纳米颗粒、(5)生理盐水+光照(1.5W/cm2、808mm)、(6)阿霉素+光照(1.5W/cm2、808mm)、(7)卟啉-硒醚多孔有机聚合物+光照(1.5W/cm2、808mm)、(8)卟啉-硒醚-阿霉素复合纳米颗粒+光照(1.5W/cm2、808mm)。通过腹腔注射(200pL)到荷瘤小鼠体内,每隔一天测量肿瘤的体积大小每天记录裸鼠的体重。
结果显示:注射卟啉-硒醚-阿霉素复合纳米颗粒后,在1.5W/cm2功率下照射5分钟,肿瘤增长比阴性对照组慢,肿瘤体积减小(图9a);在整个治疗过程中,单独静脉注射阿霉素的裸鼠体重明显下降,其他治疗组裸鼠体重基本保持不变(图9b),显示本发明卟啉-硒醚-阿霉素复合纳米颗粒在具有高效的化疗-光热联合治疗肿瘤效果的同时,还具有优良的生物相容性。
Claims (16)
1.一种卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,所述方法包括将5,10,15,20-四(4-氨基苯)-21H,23H-卟啉和二硒醚聚合,形成卟啉-硒醚多孔有机聚合物,将盐酸阿霉素包覆在所述卟啉多孔有机聚合物内,所述二硒醚为对二苯甲醛二硒醚。
2.根据权利要求1所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,所述方法包括以下步骤:
1)将盐酸阿霉素溶解于水中,得到盐酸阿霉素的水溶液;
2)将5,10,15,20-四(4-氨基苯)-21H,23H-卟啉与二硒醚通过水热反应进行聚合,真空干燥,得到卟啉-硒醚多孔有机聚合物;
将所述卟啉-硒醚多孔有机聚合物分散于水中,得到卟啉-硒醚多孔有机聚合物分散溶液;
3)将步骤2)所述卟啉-硒醚多孔有机聚合物分散溶液与所述步骤1)中盐酸阿霉素溶液混合,剧烈搅拌直至完成包覆,得到卟啉-硒醚-阿霉素复合纳米颗粒。
3.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤1)所述盐酸阿霉素的水溶液的浓度为1~10mg/mL。
4.根据权利要求3所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤1)所述盐酸阿霉素的水溶液的浓度为1mg/mL。
5.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤2)所述5,10,15,20-四(4-氨基苯)-21H,23H-卟啉与二硒醚的摩尔比为1:2。
6.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤2)水热反应的溶剂为1,4二氧六环,所述溶剂用量与5,10,15,20-四(4-氨基苯)-21H,23H-卟啉摩尔比为700:1,水热反应的条件为100℃~160℃,12~36h。
7.根据权利要求6所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤2),水热反应条件为120℃,24h。
8.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤2)所述卟啉-硒醚多孔有机聚合物分散溶液浓度为1mg/mL。
9.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤3)中所述卟啉-硒醚多孔有机聚合物分散溶液与盐酸阿霉素溶液的质量比为1:0.25~1:5。
10.根据权利要求9所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤3)所述卟啉-硒醚多孔有机聚合物分散溶液与盐酸阿霉素溶液的质量比为1:2。
11.根据权利要求2所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤3)所述搅拌时间为8-24h。
12.根据权利要求11所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法,其特征在于,步骤3)所述搅拌时间为12h;所述搅拌在黑暗环境下进行。
13.基于权利要求1~12任意一项所述的卟啉-硒醚-阿霉素复合纳米颗粒的制备方法得到的卟啉-硒醚-阿霉素复合纳米颗粒。
14.权利要求13所述的卟啉-硒醚-阿霉素复合纳米颗粒在制备靶向肿瘤药物中的应用。
15.根据权利要求14所述的应用,其特征在于,所述靶向肿瘤药物为同时具有光热治疗、化学治疗、化学动力学治疗效果的药物。
16.根据权利要求14所述的应用,其特征在于,所述靶向肿瘤药物靶向治疗前列腺癌的肿瘤药物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111475996.8A CN114099709B (zh) | 2021-12-06 | 2021-12-06 | 一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111475996.8A CN114099709B (zh) | 2021-12-06 | 2021-12-06 | 一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114099709A CN114099709A (zh) | 2022-03-01 |
CN114099709B true CN114099709B (zh) | 2023-10-27 |
Family
ID=80366766
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111475996.8A Active CN114099709B (zh) | 2021-12-06 | 2021-12-06 | 一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114099709B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115137845B (zh) * | 2022-08-30 | 2022-11-01 | 潍坊医学院附属医院 | 一种含动态亚胺键的金属有机框架共价同时固载阿霉素和卟啉复合物及其制备方法和应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101210024A (zh) * | 2006-12-30 | 2008-07-02 | 南京大学 | 中位-四芳基二菲并二硒代卟啉衍生物的合成及其应用 |
CN109223729A (zh) * | 2018-09-21 | 2019-01-18 | 华南理工大学 | 一种缩硫酮键键合阿霉素和聚磷酸酯的材料及其制备方法与应用 |
KR20210015692A (ko) * | 2019-08-02 | 2021-02-10 | 전남대학교병원 | 클로린 e6 다중결합체 및 이를 이용한 나노광증감제 |
-
2021
- 2021-12-06 CN CN202111475996.8A patent/CN114099709B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101210024A (zh) * | 2006-12-30 | 2008-07-02 | 南京大学 | 中位-四芳基二菲并二硒代卟啉衍生物的合成及其应用 |
CN109223729A (zh) * | 2018-09-21 | 2019-01-18 | 华南理工大学 | 一种缩硫酮键键合阿霉素和聚磷酸酯的材料及其制备方法与应用 |
KR20210015692A (ko) * | 2019-08-02 | 2021-02-10 | 전남대학교병원 | 클로린 e6 다중결합체 및 이를 이용한 나노광증감제 |
Non-Patent Citations (4)
Title |
---|
Antibacterial Activities of Novel Diselenide-bridged bis(Porphyrin)s on Staphylococcus aureus Investigated by Microcalorimetry;Xiang-jiao Xu et al;Biol Trace Elem Res;185-192 * |
Guanidinium-Based Ionic Covalent Organic Porous Polymer as Natamycin Delivery Agents for Anti-Candida albicans;Xia Liu et al;ChemistrySelect;第6卷;10788-10792 * |
Red light responsive diselenide-containing block copolymer micelles;Peng Han et al;J. Mater. Chem. B;第1卷;740-743 * |
共价有机多孔聚合物合成新策略;周宝龙 等;化学学报;第73卷;487-497 * |
Also Published As
Publication number | Publication date |
---|---|
CN114099709A (zh) | 2022-03-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Jiang et al. | Nanozyme catalyzed cascade reaction for enhanced chemodynamic therapy of low-H2O2 tumor | |
Yang et al. | Recent advances in nanosized metal organic frameworks for drug delivery and tumor therapy | |
Li et al. | Black phosphorus nanophototherapeutics with enhanced stability and safety for breast cancer treatment | |
CN113456816B (zh) | 一种自供氧中空普鲁士蓝纳米粒及其制备方法与应用 | |
Liu et al. | Enhancing the photodynamic therapy efficacy of black phosphorus nanosheets by covalently grafting fullerene C 60 | |
CN114099709B (zh) | 一种卟啉-硒醚-阿霉素复合纳米颗粒及其制备方法和应用 | |
CN112641946A (zh) | 聚多巴胺包裹金纳米复合物及其制备方法与在肿瘤多模态诊疗中的应用 | |
Chen et al. | Glucose oxidase-loaded colloidal stable WS2 nanobowls for combined starvation/photothermal therapy of colorectal tumors | |
CN111135313B (zh) | 一种近红外染料共配位型纳米配位聚合物及其制备方法和应用 | |
CN110790922B (zh) | 一种聚卟啉类化合物的制备方法及应用 | |
CN112206221A (zh) | 一种负载斑蝥素的巨噬细胞膜包封的金属有机框架纳米颗粒及其制备方法 | |
Xiao et al. | Boron-based nanosheets for ultrasound-mediated synergistic cancer therapy | |
Du et al. | A dual-nanozyme-loaded black phosphorus multifunctional therapeutic platform for combined photothermal/photodynamic/starvation cancer therapy | |
Xu et al. | Thermoresponsive injectable self-healing hydrogel containing polydopamine-coated Fe/Mo-doped TiO2 nanoparticles for efficient synergistic sonodynamic-chemodynamic-photothermal-chemo therapy | |
CN112656944B (zh) | 一种齐墩果酸纳米凝胶的制备方法及其应用 | |
Xiang et al. | Tumor microenviroment-responsive self-assembly of barium titanate nanoparticles with enhanced piezoelectric catalysis capabilities for efficient tumor therapy | |
Zhang et al. | Hollow mesoporous polyaniline nanoparticles with high drug payload and robust photothermal capability for cancer combination therapy | |
Jähde et al. | Protection of cultured malignant cells from mitoxantrone cytotoxicity by low extracellular pH: a possible mechanism for chemoresistance in vivo | |
CN116350800A (zh) | 葡萄糖氧化酶-金属-姜黄素自组装纳米颗粒的制备与应用 | |
Zhou et al. | Vapor deposition synthesis of polypyrrole nanoparticles with a tunable photothermal conversion capacity | |
CN113930335B (zh) | 一种纳米酶级联生物反应器及其制备方法和应用 | |
CN113769087B (zh) | 仿生预淬灭双光敏剂共组装纳米粒及其制备和应用 | |
CN110665005B (zh) | 一种掺铁聚合物纳米粒及其制备方法和应用 | |
Li et al. | Neutrophil membrane camouflaged hybrid nanozymes for enhanced starvation/photothermal tumor therapy | |
CN114028565B (zh) | 一种治疗乳腺癌的3d-cof载药体系及其制备方法 |
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 |