CN110917348A - 荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 - Google Patents
荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 Download PDFInfo
- Publication number
- CN110917348A CN110917348A CN201911247298.5A CN201911247298A CN110917348A CN 110917348 A CN110917348 A CN 110917348A CN 201911247298 A CN201911247298 A CN 201911247298A CN 110917348 A CN110917348 A CN 110917348A
- Authority
- CN
- China
- Prior art keywords
- phototherapy
- tumor
- fluorescence
- treatment agent
- targeted
- 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.)
- Pending
Links
- 238000011282 treatment Methods 0.000 title claims abstract description 50
- 238000003384 imaging method Methods 0.000 title claims abstract description 42
- 238000001126 phototherapy Methods 0.000 title claims abstract description 40
- 238000003745 diagnosis Methods 0.000 title claims abstract description 37
- 238000010276 construction Methods 0.000 title abstract description 5
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 39
- 230000008685 targeting Effects 0.000 claims abstract description 25
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 23
- 229920000642 polymer Polymers 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 20
- 229940079593 drug Drugs 0.000 claims abstract description 17
- 230000000694 effects Effects 0.000 claims abstract description 17
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 11
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 11
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 10
- 238000001338 self-assembly Methods 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 9
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 8
- 238000009825 accumulation Methods 0.000 claims abstract description 6
- 230000009977 dual effect Effects 0.000 claims abstract description 6
- 238000006069 Suzuki reaction reaction Methods 0.000 claims abstract description 5
- 229910052751 metal Inorganic materials 0.000 claims abstract description 3
- 239000002184 metal Substances 0.000 claims abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 24
- 238000000502 dialysis Methods 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 13
- 210000004027 cell Anatomy 0.000 claims description 12
- 150000003384 small molecules Chemical class 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical group CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 9
- 229920002674 hyaluronan Polymers 0.000 claims description 9
- 229960003160 hyaluronic acid Drugs 0.000 claims description 9
- 239000003504 photosensitizing agent Substances 0.000 claims description 9
- FYNROBRQIVCIQF-UHFFFAOYSA-N pyrrolo[3,2-b]pyrrole-5,6-dione Chemical compound C1=CN=C2C(=O)C(=O)N=C21 FYNROBRQIVCIQF-UHFFFAOYSA-N 0.000 claims description 8
- 229940099500 cystamine Drugs 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- OOTFVKOQINZBBF-UHFFFAOYSA-N cystamine Chemical compound CCSSCCN OOTFVKOQINZBBF-UHFFFAOYSA-N 0.000 claims description 6
- 229940039227 diagnostic agent Drugs 0.000 claims description 6
- 239000000032 diagnostic agent Substances 0.000 claims description 6
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical compound C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 claims description 5
- 230000009471 action Effects 0.000 claims description 5
- 239000005457 ice water Substances 0.000 claims description 5
- 230000005764 inhibitory process Effects 0.000 claims description 5
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 5
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 4
- -1 bromo-dithienyl pyrrolo-pyrrole-dione Chemical compound 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 229960004657 indocyanine green Drugs 0.000 claims description 4
- MOFVSTNWEDAEEK-UHFFFAOYSA-M indocyanine green Chemical compound [Na+].[O-]S(=O)(=O)CCCCN1C2=CC=C3C=CC=CC3=C2C(C)(C)C1=CC=CC=CC=CC1=[N+](CCCCS([O-])(=O)=O)C2=CC=C(C=CC=C3)C3=C2C1(C)C MOFVSTNWEDAEEK-UHFFFAOYSA-M 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- QVMPZNRFXAKISM-UHFFFAOYSA-N tirapazamine Chemical compound C1=CC=C2[N+]([O-])=NC(=N)N(O)C2=C1 QVMPZNRFXAKISM-UHFFFAOYSA-N 0.000 claims description 4
- 229950002376 tirapazamine Drugs 0.000 claims description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 3
- 239000002246 antineoplastic agent Substances 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- OYINILBBZAQBEV-UWJYYQICSA-N (17s,18s)-18-(2-carboxyethyl)-20-(carboxymethyl)-12-ethenyl-7-ethyl-3,8,13,17-tetramethyl-17,18,22,23-tetrahydroporphyrin-2-carboxylic acid Chemical compound N1C2=C(C)C(C=C)=C1C=C(N1)C(C)=C(CC)C1=CC(C(C)=C1C(O)=O)=NC1=C(CC(O)=O)C([C@@H](CCC(O)=O)[C@@H]1C)=NC1=C2 OYINILBBZAQBEV-UWJYYQICSA-N 0.000 claims description 2
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims description 2
- 125000000217 alkyl group Chemical group 0.000 claims description 2
- 229940044683 chemotherapy drug Drugs 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000003495 polar organic solvent Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 229930192474 thiophene Natural products 0.000 claims description 2
- 229940124597 therapeutic agent Drugs 0.000 claims 3
- 125000002524 organometallic group Chemical group 0.000 claims 1
- 230000005760 tumorsuppression Effects 0.000 claims 1
- 238000002604 ultrasonography Methods 0.000 claims 1
- 230000005284 excitation Effects 0.000 abstract description 8
- 238000006731 degradation reaction Methods 0.000 abstract description 2
- 238000001727 in vivo Methods 0.000 abstract description 2
- 230000014759 maintenance of location Effects 0.000 abstract 1
- 230000035699 permeability Effects 0.000 abstract 1
- 230000004614 tumor growth Effects 0.000 abstract 1
- 239000002105 nanoparticle Substances 0.000 description 14
- 239000000243 solution Substances 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000009210 therapy by ultrasound Methods 0.000 description 5
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 230000010354 integration Effects 0.000 description 4
- 239000008055 phosphate buffer solution Substances 0.000 description 4
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000002428 photodynamic therapy Methods 0.000 description 3
- 238000007626 photothermal therapy Methods 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 238000007447 staining method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000002902 bimodal effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 238000010226 confocal imaging Methods 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 125000003929 folic acid group Chemical group 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- ZDWVWKDAWBGPDN-UHFFFAOYSA-O propidium Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 ZDWVWKDAWBGPDN-UHFFFAOYSA-O 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000010837 receptor-mediated endocytosis Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- ODHXBMXNKOYIBV-UHFFFAOYSA-N triphenylamine Chemical compound C1=CC=CC=C1N(C=1C=CC=CC=1)C1=CC=CC=C1 ODHXBMXNKOYIBV-UHFFFAOYSA-N 0.000 description 1
- 125000006617 triphenylamine group Chemical group 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0052—Small organic molecules
-
- 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
- A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/221—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by the targeting agent or modifying agent linked to the acoustically-active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
- A61K49/225—Microparticles, microcapsules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- Radiology & Medical Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Acoustics & Sound (AREA)
- Biomedical Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Nanotechnology (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一种荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用，以吡咯并吡咯二酮基团为电子受体(A)，与有机电子给体基团(D)，通过有机金属催化的suzuki偶联反应，形成疏水性D‑π‑A‑π‑D结构的共轭小分子；将此共轭小分子材料接枝到亲水性的肿瘤靶向生物分子上，得到两亲性聚合物。该两亲性聚合物可通过超分子自组装形成纳米诊疗剂，可以同时负载其他抗肿瘤荧光药物。该纳米诊疗剂在体内循环时抗降解能力强，稳定性好，通过主动靶向和渗透性增强与滞留效应的双重靶向，在肿瘤部位产生药物积累，在近红外光激发下实现靶向荧光成像和光声成像，并且可以产生良好的光疗效果,有效抑制肿瘤细胞的生长。
Description
技术领域
本发明属于荧光/光声成像材料、光疗技术领域，具体涉及将D-π-A-π-D结构的共轭小分子和亲水性肿瘤靶向生物分子结合制备多功能靶向纳米诊疗剂,以及它们在荧光生物成像、光声成像、光疗一体化方面的应用。
背景技术
光疗主要包括光动力和光热治疗，即光敏剂在激光作用下，通过震动产生高温(光热)或者将能量传递给周围的氧气使其变成单线态氧(光动力)，从而破坏杀死恶性疾病细胞的一种治疗方式。光疗一体化即为在同一激发光源下实现光动力与光热联合治疗。光热治疗具有治疗时间短、治疗效率高、毒副作用低的优点，光动力治疗具有选择性好、适用性广、无耐药性等优点；由于二者结合一体化治疗可以互相促进疗效，实现无创性的高效治疗，因而被广大研究人员所关注。同时，光敏剂在发热的过程中也会产生声波，通过仪器捕获，可以实现光声成像。光声成像具有高分辨率、高对比度、组织穿透深等优点，可以实现对病变部位精确检测，从而协助治疗。
吡咯并吡咯二酮(DPP)衍生共轭小分子，具有很宽的吸收峰、很高的光热转化效率、光声特性以及一定的单线态氧产生能力，并且很容易实现近红外吸收，从而减少激发光源对于正常组织的伤害；而高的光热转化效率和单线态氧产生能力可以提供其光疗应用的潜力。DPP分子两侧连接两个富电子的π共轭桥，如噻吩基团，可以通过重原子效应增加激子系间穿越(ISC)能力，提高三线态激子含量，增加了单线态氧产生的可能。通过在其两端引入了供电子基团与DPP形成D-π-A-π-D结构的共轭小分子，供电子基团的引入可以促进吸收红移和增强电荷输运能力，而D-π-A-π-D结构大大提高了电荷的迁移速度，促使光激发电荷运转产生热量，有利于提高材料的光热转化效率。因此，基于DPP的D-π-A-π-D结构共轭小分子将有潜力作为在近红外光激发下进行光疗和光声成像的优秀光敏剂。
亲水性的肿瘤靶向生物分子如透明质酸(HA)或者连接叶酸基团的聚乙二醇，由于具有水溶性、生物相容性、肿瘤靶向性、生物可降解性和易于化学修饰等特点，可以与诊疗剂偶联形成两亲性聚合物，通过超分子自组装形成纳米粒子进入肿瘤细胞内用于成像及治疗。
通过D-π-A-π-D结构的共轭小分子与亲水性肿瘤靶生物分子偶联，可以大大提高其水溶性、生物相容性并提供了肿瘤靶向性，辅助光敏剂实现了定位光疗，避免了对正常组织的损伤，有利于提高治疗效果和光声成像的精确性。另外，利用透析法制备纳米粒子时可以进行抗肿瘤荧光药物包裹，如Ce6、ICG、TPZ等，可以提高纳米粒子的治疗能力，赋予纳米粒子近红外荧光成像能力，实现在近红外光激发下，光声和荧光成像指导光疗。
发明内容
技术问题：本发明的目的在于提出一种荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用，并且提出其在光声成像和荧光成像指导下光动力光热联合治疗中的应用。该类荧光/光声成像及光疗一体化靶向纳米诊疗剂旨在通过主动靶向和EPR效应实现双重靶向输送药物，在肿瘤部位产生药物积累，在近红外激光作用下进行靶向荧光成像和光声成像双模态成像，同时指导光动力光热联合治疗，从而极大地提升纳米诊疗剂的诊断和治疗效果。
技术方案：本发明的一种荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用，将疏水性的D-π-A-π-D结构的共轭小分子材料接枝到亲水性的肿瘤靶向生物分子上，得到两亲性聚合物；该两亲性聚合物可通过超分子自组装形成纳米诊疗剂，可以同时负载其他抗肿瘤荧光药物；该纳米诊疗剂在体内循环时抗降解能力强，稳定性好，通过主动靶向和EPR效应的双重靶向，在肿瘤部位产生药物积累，在近红外激光作用下实现靶向荧光成像和光声成像，产生良好的光疗效果，体现出良好的肿瘤细胞抑制能力。
所述的D-π-A-π-D结构的共轭小分子具有如下分子式：
其中，两端取代的D基团为电子给体基团,噻吩为π共轭桥，吡咯并吡咯二酮(DPP)为电子受体，简称A，吡咯并吡咯二酮侧链R基团为烷基链，组成一种D-π-A-π-D的共轭结构。
所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂，所述的两亲性聚合物中，作为优选方案，D基团为供电子的三苯胺(TPA)基团，R为叔丁基，D-π-A-π-D结构的共轭小分子其结构式如下:
相应的，作为优选方案，可选择亲水性肿瘤靶向生物分子为透明质酸(HA)，两亲性的聚合物结构式如下：
上述的一种荧光/光声成像及光疗一体化靶向纳米诊疗剂的制备方法，包括以下步骤：
1)将D-π-A-π-D结构的共轭小分子接枝到胱胺修饰的HA上，制备两亲性聚合物；
2)将上述两亲性聚合物与疏水性抗肿瘤荧光药物混合，通过透析法自组装制备得到疏水空腔中包裹药物的生物纳米诊疗剂。
其中，该制备方法中的透析法具体为：
将两亲性的聚合物溶于极性有机溶剂二甲基亚砜(DMSO)中，然后将DMSO溶液在超声状态下滴加到纯水中(体积比率为1:1)，搅拌均匀，接着在冰水浴中用尖头型超声仪间隔性超声10min(超声功率为55％，每超声10s间隔2s)，室温搅拌3小时后移入透析袋中，在纯水中透析24小时，用0.45μm滤孔的滤膜过滤后，取滤液冷冻干燥，得到靶向纳米诊疗剂。
作为优选方案，所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂HA-TTDTT的制备方法为：
1)采用有机金属催化的suzuki偶联反应，将4-硼酸三苯胺接枝到溴代二噻吩基吡咯并吡咯二酮两侧，得到D-π-A-π-D结构的共轭小分子TTDTT，然后通过侧链脱叔丁酯得到可修饰的含羧酸钠的侧链。
2)由步骤1)制备得到的D-π-A-π-D结构的共轭小分子TTDTT在1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)催化下，接枝到胱胺修饰的肿瘤靶向生物分子HA上，得到两亲性聚合物。
上述的光疗一体化靶向纳米诊疗剂的应用，所述靶向生物纳米诊疗剂应用在肿瘤靶向细胞成像和光疗中，还可以用于包裹(吲哚菁绿(ICG)、二氢卟吩e6(Ce6)或替拉扎明(TPZ))等光敏剂或者化疗药物，提高材料的肿瘤抑制能力和提供荧光成像能力。该应用包括以下步骤：
1)将D-π-A-π-D结构的共轭小分子接枝到胱胺修饰的HA上，制备两亲性聚合物；
2)将上述两亲性聚合物与疏水性抗肿瘤荧光药物混合，通过透析法自组装制备得到疏水空腔中包裹药物的生物纳米诊疗剂；
3)将上述靶向纳米诊疗剂应用于肿瘤细胞，通过主动靶向和EPR效应的双重靶向，在肿瘤部位产生药物积累，实现靶向荧光成像和光声成像，在近红外光作用下可以产生良好的光疗效果和肿瘤细胞抑制能力。
有益效果：在本发明中，通过超分子自组装制备得到一种荧光/光声成像及光疗一体化靶向纳米诊疗剂。其中，自组装形成纳米诊疗剂之后，通过主动肿瘤靶向和EPR效应双重靶向，多功能的纳米诊疗剂能够稳定地输送到肿瘤部位，再通过HA受体介导的内吞作用选择性地被肿瘤细胞摄取，从而提高细胞内药物积累和增强抗肿瘤功效；并且 D-π-A-π-D结构的共轭小分子TTDTT的光声信号增强，负载的抗肿瘤药物荧光加强，达到肿瘤靶向荧光/光声成像和引导光疗的目的，从而使这种多种功能的纳米诊疗剂在荧光/光声成像和光疗一体化方面有着潜在的广泛应用前景。
附图说明
图1为本发明制备的光声成像及光疗一体化靶向纳米诊疗剂HA-TTDTT纳米粒子(NP)的结构示意图。
图2为本发明制备的负载光敏剂Ce6的荧光/光声成像及光疗一体化靶向纳米诊疗剂Ce6@HA-TTDTT NP的结构示意图。
图3为本发明制备的荧光/光声成像及光疗一体化靶向纳米诊疗剂Ce6@HA-TTDTTNP在水中的紫外吸收与荧光发射光谱。
图4为本发明制备的荧光/光声成像及光疗一体化靶向纳米诊疗剂Ce6@HA-TTDTTNP的TEM图。
图5为本发明制备的光声成像及光疗一体化靶向纳米诊疗剂HA-TTDTT NP在水溶液中的光声信号(PA signal)测试。
图6为本发明制备的荧光/光声成像及光疗一体化靶向纳米诊疗剂Ce6@HA-TTDTTNP 在肿瘤细胞HeLa中的激光共聚焦荧光显微镜近红外的细胞成像(激发波长：635nm，收集波长范围：700nm-800nm)。
图7为本发明制备的荧光/光声成像及光疗一体化靶向纳米诊疗剂Ce6@HA-TTDTTNP 在肿瘤细胞HeLa中，进行钙黄绿素/碘化丙啶细胞染色法(AM/PI染色法)测试光疗一体化对HeLa细胞的抑制效果。
具体实施方式
为了更好地理解本发明的内容，下面通过具体的实施例进一步说明本发明的技术方案，但不能以此来限制本发明的范围。
实施例：
1)通过下图方法制备，将(1)和(2)按物质的量比为2:1投料，然后在Pd[P(C6H5)3]4催化下通过Suzuki反应将二者偶联起来，反应24小时后，即得到D-π-A-π-D结构的共轭小分子；进一步用三氟乙酸进行去叔丁酯(BOC)反应，反应4小时，得到TTDTT。
2)通过下图方法制备透明质酸-胱胺(HA-Cystamine)后，再将TTDTT溶于DMSO 中，将催化剂EDC和NHS分别溶解在少量磷酸盐缓冲液(PBS)中，加至TPO上的DMSO 溶液中，反应2小时，再加入2-巯基乙醇猝灭EDC的反应活性，反应10分钟后，滴加一定量的HA-Cystamine(事先溶于少量的PBS溶液，再加入DMSO溶液，比率为1:15)，之后在室温条件下搅拌过夜。将反应瓶中的溶液移入截留分子量(MWCO)3500Da的透析袋中，透析除去未反应的EDC,NHS,2-巯基乙醇，未反应的TTDTT和其他中间产物，之后冷冻干燥36小时，获得两亲性聚合物HA-TTDTT。
3)将步骤2)中制备的HA-TTDTT溶于DMSO溶液中，在冰水浴中用尖头型超声仪超声使其尽量溶解，然后滴加到等体积的纯水中，之后继续在冰水浴中用尖头型超声仪超声10min，搅拌3小时后移入MWCO为3500Da的透析袋中，在去离子水中透析24小时，期间换水3次，之后用0.45μm滤膜过滤并冷冻干燥，得到靶向纳米粒子HA-TTDTT NP。
4)将光敏剂Ce6溶解于DMSO中，在超声状态下滴加到溶有步骤2)中制备的HA-TTDTT 的DMSO/H20混合溶液中，之后在冰水浴中用尖头型超声仪超声10分钟，搅拌2小时后移入MWCO为3500Da的透析袋中，在去离子水中透析24小时,之后用0.45μm滤膜过滤并冷冻干燥，最终得到负载Ce6的靶向纳米探针Ce6@HA-TTDTT。
5)用含有靶向纳米探针Ce6@HA-TTDTT NP的培养基孵育过量表达CD44受体的肿瘤细胞HeLa，4小时后将细胞置于激光共聚焦荧光显微镜下，以635nm的激光激发，收集700-800nm之间的荧光，拍摄照片，获得细胞成像结果。
6)以HeLa细胞为模型进行光热、光动力协同治疗效果检测，先前步骤与纳米粒子的共聚焦荧光成像相同，玻璃底的共聚焦皿在培养箱中培养24h后。吸取上清液，分两组加入用不完全培养液DMEM配置浓度为20μΜ的HA-TTDTT NP和 Ce6@HA-TTDTT NP溶液，孵育4h，用PBS缓冲液轻微清洗细胞2遍后，进行 PTT/PDT，用635nm激光器光照(0.8W/cm2，8min)，之后加入1mL DMEM包含的AM/PI指示剂(5×10-6M)。一段时间后通过共聚焦成像，PI信号采用515nm的激光作为激发光源，收集600-650nm波段的发射信号。AM信号采用488nm激光作为激发光源，收集500-550nm波段发射信号，并叠加成图。
以上所述仅是本发明的优选实施方式，应当指出：对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (8)
1.一种荧光/光声成像及光疗一体化靶向纳米诊疗剂，其特征在于，由疏水性的D-π-A-π-D结构的共轭小分子材料接枝到亲水性的肿瘤靶向生物分子上得到的两亲性聚合物通过超分子自组装形成该纳米诊疗剂，所述的D-π-A-π-D结构的共轭小分子具有如下分子式：
其中，两端取代的D基团为电子给体基团，噻吩为π共轭桥，吡咯并吡咯二酮DPP为电子受体，吡咯并吡咯二酮侧链R基团为烷基链，组成一种D-π-A-π-D的共轭结构。
2.根据权利要求1所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂，其特征在于，所述D基团为供电子的三苯胺TPA基团，R为叔丁基，D-π-A-π-D结构的共轭小分子TTDTT其结构式如下:
所述亲水性肿瘤靶向生物分子为透明质酸HA，两亲性的聚合物HA-TTDTT结构式如下：
3.如权利要求1或2所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂的构建方法，其特征在于，包括以下步骤：
1)将D-π-A-π-D结构的共轭小分子接枝到胱胺修饰的HA上，制备两亲性聚合物；
2)将上述两亲性聚合物与疏水性抗肿瘤荧光药物混合，通过透析法自组装制备得到疏水空腔中包裹药物的生物纳米诊疗剂。
4.一种如权利要求3所述的一种荧光/光声成像及光疗一体化靶向纳米诊疗剂的构建方法，其特征在于，步骤2)中，所述透析法具体为：
将两亲性的聚合物溶于极性有机溶剂二甲基亚砜,DMSO中,然后将DMSO溶液在超声状态下滴加到纯水中，体积比率为1:1搅拌均匀；接着在冰水浴中用尖头型超声仪间隔性超声10min，超声功率为55％，每超声10s间隔2s；室温搅拌3小时后移入透析袋中，在纯水中透析24小时；用0.45μm滤孔的滤膜过滤后，取滤液冷冻干燥，得到靶向纳米诊疗剂。
5.根据权利要求3所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂的构建方法，其特征在于，步骤1)中，所述D-π-A-π-D结构的共轭小分子材料是以吡咯并吡咯二酮基团为电子受体A，与有机电子给体基团D通过有机金属催化的suzuki偶联反应形成。
6.根据权利要求3或5所述的荧光/光声成像及光疗一体化靶向纳米诊疗剂的构建方法，其特征在于，步骤1)中，所述的两亲性聚合物的制备方法为：
1-1)采用有机金属催化的suzuki偶联反应，将4-硼酸三苯胺接枝到溴代二噻吩基吡咯并吡咯二酮两侧，得到D-π-A-π-D结构的共轭小分子TTDTT，然后通过侧链脱叔丁酯得到可修饰的含羧酸钠的侧链；
1-2)由步骤1-1)制备得到的D-π-A-π-D结构的共轭小分子TTDTT在1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐EDC和N-羟基琥珀酰亚胺NHS催化下，接枝到胱胺修饰的肿瘤靶向生物分子HA上，得到两亲性聚合物。
7.一种如权利要求1或2所述的光疗一体化靶向纳米诊疗剂的应用，其特征在于，所述靶向生物纳米诊疗剂应用在肿瘤靶向细胞成像和光疗中，及用于包裹包括吲哚菁绿ICG、二氢卟吩e6或替拉扎明TPZ)在内的光敏剂或者化疗药物，提高材料的肿瘤抑制能力和提供荧光成像能力。
8.一种如权利要求7所述的光疗一体化靶向纳米诊疗剂的应用，其特征在于，该应用包括以下步骤：
1)将D-π-A-π-D结构的共轭小分子接枝到胱胺修饰的HA上，制备两亲性聚合物；
2)将上述两亲性聚合物与疏水性抗肿瘤荧光药物混合，通过透析法自组装制备得到疏水空腔中包裹药物的生物纳米诊疗剂；
3)将上述靶向纳米诊疗剂应用于肿瘤细胞，通过主动靶向和EPR效应的双重靶向，在肿瘤部位产生药物积累，实现靶向荧光成像和光声成像，在近红外光作用下产生光疗效果和肿瘤细胞抑制能力。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911247298.5A CN110917348A (zh) | 2019-12-09 | 2019-12-09 | 荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911247298.5A CN110917348A (zh) | 2019-12-09 | 2019-12-09 | 荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110917348A true CN110917348A (zh) | 2020-03-27 |
Family
ID=69858329
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911247298.5A Pending CN110917348A (zh) | 2019-12-09 | 2019-12-09 | 荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110917348A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704685A (zh) * | 2021-01-21 | 2021-04-27 | 南通大学 | 一种顺铂配位体及其在制备肿瘤纳米诊疗剂中的应用 |
| CN113773667A (zh) * | 2021-09-22 | 2021-12-10 | 南京邮电大学 | 有机小分子近红外二区荧光染料及其制备方法和应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108210938A (zh) * | 2017-12-26 | 2018-06-29 | 南京邮电大学 | 一种多功能靶向纳米荧光探针及其制备与应用 |
| CN108714222A (zh) * | 2018-05-31 | 2018-10-30 | 南京邮电大学 | 一种新型多功能超分子囊泡的制备与应用 |
| CN108864106A (zh) * | 2018-07-17 | 2018-11-23 | 南京邮电大学 | 新型近红外二区有机小分子荧光探针的制备与应用 |
-
2019
- 2019-12-09 CN CN201911247298.5A patent/CN110917348A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108210938A (zh) * | 2017-12-26 | 2018-06-29 | 南京邮电大学 | 一种多功能靶向纳米荧光探针及其制备与应用 |
| CN108714222A (zh) * | 2018-05-31 | 2018-10-30 | 南京邮电大学 | 一种新型多功能超分子囊泡的制备与应用 |
| CN108864106A (zh) * | 2018-07-17 | 2018-11-23 | 南京邮电大学 | 新型近红外二区有机小分子荧光探针的制备与应用 |
Non-Patent Citations (1)
| Title |
|---|
| 苏哲等: "近红外二区荧光探针在生物成像领域的研究进展", 《应用化学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704685A (zh) * | 2021-01-21 | 2021-04-27 | 南通大学 | 一种顺铂配位体及其在制备肿瘤纳米诊疗剂中的应用 |
| CN112704685B (zh) * | 2021-01-21 | 2021-09-14 | 南通大学 | 一种顺铂配位体及其在制备肿瘤纳米诊疗剂中的应用 |
| CN113773667A (zh) * | 2021-09-22 | 2021-12-10 | 南京邮电大学 | 有机小分子近红外二区荧光染料及其制备方法和应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wen et al. | One-for-all phototheranostics: Single component AIE dots as multi-modality theranostic agent for fluorescence-photoacoustic imaging-guided synergistic cancer therapy | |
| Yin et al. | Organic semiconducting polymer amphiphile for near-infrared-II light-triggered phototheranostics | |
| Tian et al. | Recent advances of multi-dimensional porphyrin-based functional materials in photodynamic therapy | |
| Zheng et al. | Biodegradable hypocrellin derivative nanovesicle as a near-infrared light-driven theranostic for dually photoactive cancer imaging and therapy | |
| Li et al. | Recent advances in the development of NIR-II organic emitters for biomedicine | |
| Su et al. | A supramolecular strategy to engineering a non-photobleaching and near-infrared absorbing nano-J-aggregate for efficient photothermal therapy | |
| Jun et al. | Folic acid–conjugated chitosan-functionalized graphene oxide for highly efficient photoacoustic imaging-guided tumor-targeted photothermal therapy | |
| Gulzar et al. | Nano-graphene oxide-UCNP-Ce6 covalently constructed nanocomposites for NIR-mediated bioimaging and PTT/PDT combinatorial therapy | |
| Feng et al. | A versatile near infrared light triggered dual-photosensitizer for synchronous bioimaging and photodynamic therapy | |
| Li et al. | Core-satellite metal-organic framework@ upconversion nanoparticle superstructures via electrostatic self-assembly for efficient photodynamic theranostics | |
| CN113087877B (zh) | 近红外二区荧光发射水溶性共轭聚合物纳米光疗试剂及其制备方法与应用 | |
| Huang et al. | Hyaluronic acid nanoparticles based on a conjugated oligomer photosensitizer: Target-specific two-photon imaging, redox-sensitive drug delivery, and synergistic chemo-photodynamic therapy | |
| CN104162164A (zh) | 一种包含吲哚青绿的多聚体白蛋白纳米球及其制备方法和应用 | |
| CN109503455B (zh) | 一种以四苯乙烯吲哚衍生物为光敏剂的纳米复合材料及制备方法和在肿瘤治疗方面的应用 | |
| Deng et al. | Zwitterionic diketopyrrolopyrrole for fluorescence/photoacoustic imaging guided photodynamic/photothermal therapy | |
| Deng et al. | 808 nm light responsive nanotheranostic agents based on near-infrared dye functionalized manganese ferrite for magnetic-targeted and imaging-guided photodynamic/photothermal therapy | |
| Yang et al. | A Nd 3+ sensitized upconversion nanosystem with dual photosensitizers for improving photodynamic therapy efficacy | |
| Battogtokh et al. | Glycol chitosan-coated near-infrared photosensitizer-encapsulated gold nanocages for glioblastoma phototherapy | |
| Liu et al. | Thiophene donor for NIR-II fluorescence imaging-guided photothermal/photodynamic/chemo combination therapy | |
| Shu et al. | Enhanced photothermal performance by carbon dot-chelated polydopamine nanoparticles | |
| Li et al. | Degradable multifunctional porphyrin-based porous organic polymer nanosonosensitizer for tumor-specific sonodynamic, chemo-and immunotherapy | |
| Zhang et al. | Novel Donor–Acceptor Conjugated Polymer-Based Nanomicelles for Photothermal Therapy in the NIR Window | |
| CN110917348A (zh) | 荧光/光声成像及光疗一体化靶向纳米诊疗剂及其构建与应用 | |
| CN111249461A (zh) | 藻蓝蛋白-二氢卟酚e6共价纳米颗粒的制备及其应用 | |
| Li et al. | Innovative design strategies advance biomedical applications of phthalocyanines |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200327 |
|
| RJ01 | Rejection of invention patent application after publication |