CN114181253B - 一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 - Google Patents
一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 Download PDFInfo
- Publication number
- CN114181253B CN114181253B CN202111476712.7A CN202111476712A CN114181253B CN 114181253 B CN114181253 B CN 114181253B CN 202111476712 A CN202111476712 A CN 202111476712A CN 114181253 B CN114181253 B CN 114181253B
- Authority
- CN
- China
- Prior art keywords
- mito
- mitochondria
- application
- mitochondrial
- 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.)
- Active
Links
- 239000003504 photosensitizing agent Substances 0.000 title claims abstract description 19
- 230000008685 targeting Effects 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 210000003470 mitochondria Anatomy 0.000 title claims description 12
- 238000002428 photodynamic therapy Methods 0.000 claims abstract description 12
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 5
- 230000002438 mitochondrial effect Effects 0.000 claims abstract description 4
- 239000003814 drug Substances 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 4
- 201000010099 disease Diseases 0.000 claims 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 2
- 230000001225 therapeutic effect Effects 0.000 claims 1
- 206010034972 Photosensitivity reaction Diseases 0.000 abstract description 7
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Substances C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000036211 photosensitivity Effects 0.000 abstract description 4
- -1 triphenylphosphine cation Chemical class 0.000 abstract description 4
- 238000005886 esterification reaction Methods 0.000 abstract description 3
- 210000001700 mitochondrial membrane Anatomy 0.000 abstract description 3
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 238000001179 sorption measurement Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 24
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 15
- 239000003642 reactive oxygen metabolite Substances 0.000 description 10
- 239000000523 sample Substances 0.000 description 9
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 8
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000011534 incubation Methods 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 210000003463 organelle Anatomy 0.000 description 5
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 4
- 230000006907 apoptotic process Effects 0.000 description 4
- 230000008045 co-localization Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- ZKSVYBRJSMBDMV-UHFFFAOYSA-N 1,3-diphenyl-2-benzofuran Chemical compound C1=CC=CC=C1C1=C2C=CC=CC2=C(C=2C=CC=CC=2)O1 ZKSVYBRJSMBDMV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 208000007578 phototoxic dermatitis Diseases 0.000 description 3
- 231100000018 phototoxicity Toxicity 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000002132 lysosomal effect Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 102000011727 Caspases Human genes 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 102100030497 Cytochrome c Human genes 0.000 description 1
- 108010075031 Cytochromes c Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 231100000991 Reactive Oxygen Species (ROS) Photosafety Assay Toxicity 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 208000006593 Urologic Neoplasms Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000004103 aerobic respiration Effects 0.000 description 1
- 238000004873 anchoring Methods 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000010001 cellular homeostasis Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000007810 chemical reaction solvent Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000027829 mitochondrial depolarization Effects 0.000 description 1
- 230000004065 mitochondrial dysfunction Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000010627 oxidative phosphorylation Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000006552 photochemical reaction Methods 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000005381 potential energy Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000010282 redox signaling Effects 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/54—Quaternary phosphonium compounds
- C07F9/5442—Aromatic phosphonium compounds (P-C aromatic linkage)
-
- 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
- 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
-
- 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
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1014—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
- C09K2211/1055—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms with other heteroatoms
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明提供了一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用。基于重原子引入具有增强光敏性的BODIPY染料(BD)，通过酯化反应将广泛认可的具有线粒体靶向的三苯基膦阳离子(TPP)衍生在BODIPY核的头部得到mitoBD。利用TPP和线粒体膜电位的静电吸附作用以及脂溶性，mitoBD可以选择性精准地靶向到细胞器‑线粒体，进而增强对肿瘤细胞的光动力治疗效果，同时具有荧光成像的应用潜力。
Description
技术领域
本发明属于化学生物学领域，研究内容是基于三苯基膦共价修饰的氟硼二吡咯(BODIPY)染料分子，通过靶向细胞器-线粒体增强对肿瘤细胞的光动力治疗效果，同时具有荧光成像的应用潜力。
背景技术
光动力治疗(PDT)是一种众所周知的广泛用于治疗各种癌症的非侵入性疗法，尤其是对膀胱癌、泌尿道癌、肺癌和食管癌。PDT的作用机制是通过光敏剂、光和氧气相互作用引发光化学反应产生活性氧(ROS)，进而触发一系列反应杀伤癌细胞：(1)活性氧能够氧化细胞内的生物大分子直接引起细胞凋亡或者坏死；(2)它还能诱导局部的炎症反应，激活靶细胞的免疫反应从而对微脉管***产生损伤，间接杀伤靶细胞。然而，1O2的寿命很短(≈3.0μs)并且扩散半径有限(0.02μm)，只会对其附近的细胞构造产生光毒性。因此，如何将光敏剂精确地运输到重要或者较为脆弱细胞器对于提高光动力的治疗效果尤其重要。
随着对光动力机制认识的深入，人们逐渐开发靶向特定细胞器(细胞膜、细胞核、内质网、溶酶体和线粒体)的光敏剂用于对抗肿瘤。目前，细胞核靶向的光动力治疗虽然对癌细胞有很大的杀伤性，但同时伴有遗传变异的风险。考虑到光敏剂小分子脂溶性以及细胞膜不可避免地快速摄取特性，实现长时间的锚定在细胞膜上仍然是一个挑战。溶酶体靶向的光动力疗法并没有像其它靶向疗法那样引起人们的兴趣，主要是因为含有蛋白质和多肽的治疗剂对溶酶体微环境不具有耐受性。内质网靶向的光敏剂目前研究较少，相关的靶向作用机制尚不清楚，需要进一步探索。相比于前面那些缺陷，线粒体因其独特结构特点以及在细胞中重要功能使其在细胞器靶向的PDT中脱颖而出，进而受到了科学家们越来越多的关注。
线粒体是在大多数细胞中含有的由两层膜包被的细胞器，它负责有氧呼吸的第二和第三阶段，即三羧酸循环与氧化磷酸化，是细胞进行产生ATP的主要场所，因而被称为“power house”。同时，线粒体是大部分ROS产生的主要场所，它拥有一系列维持癌细胞稳态的调节***，在调节氧化还原信号、细胞凋亡以及坏死途径中起重要作用。当线粒体中ROS过表达时，表现为氧化应激增加，进而发生线粒体膜去极化以及释放细胞色素c和激活半胱天冬酶等细胞凋亡过程。此外，线粒体自身结构特点较为突出：外膜具有高渗透性，而内膜是不可渗透的。内膜上分布的电子传递链在产生能量时会将电化学势能存储，在内膜两侧造成质子及其他离子浓度的不对称分布而形成远远高于其他细胞器的线粒体膜电位(MMP，-180mV)。因此，线粒体是PDT的理想靶标细胞器，它可以切断细胞的能量供应并导致线粒体功能障碍引发细胞凋亡。
发明内容
本发明提供了一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用。基于重原子引入具有增强光敏性的BODIPY染料(BD)，通过酯化反应将广泛认可的具有线粒体靶向的三苯基膦阳离子(TPP)衍生在BODIPY核的头部得到mitoBD。利用TPP和线粒体膜电位的静电吸附作用以及脂溶性，mitoBD可以选择性精准地靶向到细胞器-线粒体，进而增强对肿瘤细胞的光动力治疗效果，同时具有荧光成像的应用潜力。
本发明提供的mitoBD分子，其结构式如图1所示
本发明所提供合成方法如图2所示，通过酯化反应，以二氯甲烷作为反应溶剂，在1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDCI，0.10mmol)和4-二甲基氨基吡啶(DMAP，0.10mmol)催化下，分别加入BD和三苯基膦(0.95g，0.42g)于圆底烧瓶中，常温搅拌反应6小时，即得mitoBD。
本发明所提供了mitoBD分子的紫外-可见光吸收光谱，如图3所示：当mitoBD处于有机溶液中以单分子形式存在时，其最大吸收峰在678nm。
本发明所提供了mitoBD分子荧光性质的考察结果，如图4所示：当mitoBD处于有机溶液中以单分子形式存在时，其可以发射红色荧光，最大发射峰在723nm。
本发明所提供了mitoBD分子光敏性的考察结果，如图5(A)和5(B)所示：使用DPBF作为检测ROS的探针，评价mitoBD分子在黑暗以及655nm激光下照射90秒活性氧的产生，每间隔15秒利用紫外分光光度计扫描记录探针吸收变化。在90秒内，DPBF在413nm的吸收从1.0降至0.2，表明mitoBD分子具有作为光敏剂的潜能。
选用HeLa细胞，利用Leica TCS SP8共焦激光扫描仪对mitoBD分子在细胞内的荧光以及共定位进行了考察，结果如图6所示：mitoBD分子可以发射明亮的红色荧光，具有成像诊断的应用潜力。同时选用商用的线粒体探针 Green FM根据相关操作进行了共定位实验，mitoBD分子与线粒体皮尔森相关系数达到0.91，即实现了很好的共定位。
选用HeLa细胞，利用DCFH-DA探针和MTT法分别检测mitoBD分子在细胞内ROS产生以及抑制癌细胞生长的情况，结果如图7(A)和7(B)所示：同黑暗条件相比，光照组mitoBD分子在655nm激光下可以产生明显的ROS，进而产生光毒性杀死癌细胞。
附图说明
图1为本发明中所合成的化合物分子结构式。
图2为本发明中所设计分子的合成路线。
图3为本发明中mitoBD分子在DMSO中的紫外可见吸收光谱。
图4为本发明中mitoBD分子在DMSO中的荧光发射图谱。
图5为本发明中mitoBD分子在黑暗以及光照下活性氧的产生情况。
图6为本发明中mitoBD分子在细胞中荧光以及与线粒体共定位的情况。
图7为本发明中mitoBD分子在细胞中产生活性氧情况以及MTT实验的结果。
具体实施方式
下面结合具体实施例对本发明做出进一步的解释和说明，基于本发明中的实施例，本领域的技术人员，在没有做出创造性的劳动成果下所获得的其他实施例，都属于本发明的保护范围。
实施例1。光敏剂氟硼二吡咯-三苯基膦(mitoBD)分子的合成，合成路线见图2：向化合物BD(0.95g，1.0mmol)和三苯基膦溴丁酸(0.42g，1.0mmol)的二氯甲烷混合溶液中加入1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDCI，0.10mmol)和4-二甲基氨基吡啶(DMAP，0.10mmol)，将混合物在室温避光条件下搅拌过夜。待反应完全后停止反应，将所得的混合物用饱和盐水洗涤三次，无水硫酸钠干燥，减压下旋蒸除去溶剂。将粗产物在硅胶柱上进行纯化分离，以二氯甲烷/甲醇＝100/4作为洗脱剂，得到深绿色固体0.54g，命名为mitoBD，产率为40.5％。1H NMR(400MHz,CDCl3)δ(ppm)8.08(d,J＝16.5Hz,2H),7.93–7.84(m,6H),7.79(m,3H),7.70(m,6H),7.60(d,J＝16.5Hz,2H),7.24(d,J＝8.4Hz,2H),7.16(s,2H),6.91(d,J＝8.4Hz,2H),6.76(s,2H),4.23(m,2H),4.17(m,2H),4.07(m,2H),3.97(s,6H),3.93(s,6H),3.73(m,6H),2.96(m,2H),2.09(s,6H),1.93(m,4H),1.46(s,6H).13C NMR(100MHz,CDCl3)δ(ppm)173.16,150.45,149.20,148.17,140.33,138.86,136.98,135.00,134.97,133.83,133.73,131.63,130.50,130.38,130.16,121.64,118.63,117.77,116.41,114.48,111.23,109.86,70.81,70.65,69.76,68.95,67.35,63.93,56.03,55.98,20.03,18.03,12.65.HRMS(ESI):m/z[M+NH4]+calcd.for C67H69BBr2F2N2O9P 1285.3148；found1285.3155.
实施例2。对mitoBD分子吸收以及荧光性质的研究：通过紫外-可见吸收分光光度计和荧光分光光度计(λEx＝676nm)测量mitoBD分子在DMSO中的荧光光谱。结果如图3和图4所示。
实施例3。对mitoBD分子光敏性的研究：通常，将DPBF探针溶液稀释添加到2mLmitoBD/DMSO溶液中并置于比色皿，同时确保探针在412nm处的吸收为1.0，然后用655nm激光照射90秒。每隔15秒记录探针在412nm处吸光度的变化。mitoBD分子在黑暗以及光照下产生1O2结果如图5(A)和5(B)所示。
实施例4。对mitoBD分子在细胞内荧光成像以及共定位进行研究。将HeLa细胞(密度约1×105)接种到规格为20mm的共聚焦细胞培养皿中，置于细胞培养箱(5％CO2，37℃)孵育过夜。待细胞完全贴壁除去旧培养液，然后用药物继续孵育。随后在避光的环境下，加入1mLMito-tracker探针溶液(浓度为200nM)，置于培养箱染色30分钟。然后将细胞用PBS(10mM，pH 7.4)洗涤几次，通过Leica TCS SP8共焦激光扫描仪拍摄(mitoBD：λEx＝633nm，λEm＝675-735nm，Tracker：λEx＝488nm，λEm＝500-580nm)，实验结果如图6所示。
实施例5。为了测定细胞内活性氧产生以及对癌细胞的抑制情况。首先选用DCFH-DA探针进行ROS测定。铺板以及给药孵育时间同上，随后，在避光的环境下加入新鲜配置的DCFH-DA探针溶液(1mL，浓度为20μM)，置于培养箱共同孵育30分钟。洗涤几次并置于黑暗或者655nm激光下照射1分钟。最后,通过在倒置荧光显微分别在明场和荧光场下成像，实验结果如图7(A)所示。其次，利用MTT法进行光毒性测定。将HeLa细胞(密度约5×103)接种到96孔板中，置于细胞培养箱(5％CO2，37℃)孵育过夜。待细胞完全贴壁除去旧培养液，然后用不同浓度的药物继续孵育。最后，洗涤几次并置于黑暗或者655nm激光下照射1分钟，37℃继续孵育，随后加入MTT，37℃孵育4小时后加入DMSO，用酶标仪测试96孔板的吸收值。实验结果如图7(B)所示。
以上所述仅是本发明的优选实施方式，应当指出，对于使用本技术领域的研究人员，在不脱离本发明技术原理的前提下，对这些实施例进行的多种修改，这些修改应该视为本发明的保护范围。
Claims (5)
1.一种靶向线粒体的光敏剂mitoBD，其特征在于，结构式如下：
2.一种根据权利要求1所述的靶向线粒体的光敏剂mitoBD的制备方法，其特征在于，制备路线如下：
3.一种靶向线粒体的光敏剂mitoBD的应用，其特征在于，将权利要求1所述的靶向线粒体的光敏剂mitoBD应用于制备光动力治疗药物。
4.一种靶向线粒体的光敏剂mitoBD的应用，其特征在于，将权利要求1所述的靶向线粒体的光敏剂mitoBD应用于制备靶向线粒体的光动力治疗药物。
5.一种靶向线粒体的光敏剂mitoBD的应用，其特征在于，将权利要求1所述的靶向线粒体的光敏剂mitoBD应用于非疾病诊断目的或非疾病治疗目的的荧光成像。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111476712.7A CN114181253B (zh) | 2021-12-06 | 2021-12-06 | 一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111476712.7A CN114181253B (zh) | 2021-12-06 | 2021-12-06 | 一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114181253A CN114181253A (zh) | 2022-03-15 |
| CN114181253B true CN114181253B (zh) | 2023-10-31 |
Family
ID=80542402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111476712.7A Active CN114181253B (zh) | 2021-12-06 | 2021-12-06 | 一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114181253B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115006527B (zh) * | 2022-05-20 | 2023-03-17 | 苏州大学 | 一种线粒体靶向光敏剂及其制备方法和应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109054807A (zh) * | 2018-08-27 | 2018-12-21 | 华南理工大学 | 一种双细胞器靶向的纳米探针及其制备及应用 |
| CN111848658A (zh) * | 2020-06-17 | 2020-10-30 | 浙江理工大学 | 一种靶向线粒体的氟硼二吡咯类化合物及其脂质体包裹纳米粒子的制备方法和用途 |
-
2021
- 2021-12-06 CN CN202111476712.7A patent/CN114181253B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109054807A (zh) * | 2018-08-27 | 2018-12-21 | 华南理工大学 | 一种双细胞器靶向的纳米探针及其制备及应用 |
| CN111848658A (zh) * | 2020-06-17 | 2020-10-30 | 浙江理工大学 | 一种靶向线粒体的氟硼二吡咯类化合物及其脂质体包裹纳米粒子的制备方法和用途 |
Non-Patent Citations (2)
| Title |
|---|
| A Photosensitive Polymeric Carrier with a Renewable Singlet Oxygen Reservoir Regulated by Two NIR Beams for Enhanced Antitumor Phototherapy;Yang Chun 等;Small;第17卷(第29期);第1-12页 * |
| A Photostable Near-Infrared Fluorescent Tracker with pH-Independent Specificity to Lysosomes for Long Time and Multicolor Imaging;Zhang Xinfu 等;ACS Applied Materials & Interfaces;第06卷(第23期);第21669-21676页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114181253A (zh) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhou et al. | Enhancing the ROS generation ability of a rhodamine-decorated iridium (III) complex by ligand regulation for endoplasmic reticulum-targeted photodynamic therapy | |
| Dai et al. | A cell-membrane-permeable europium complex as an efficient luminescent probe for singlet oxygen | |
| Wang et al. | Discrimination between cancerous and normal cells/tissues enabled by a near-infrared fluorescent HClO probe | |
| CN111875604B (zh) | 一类线粒体靶向和光动力治疗的β-咔啉鎓盐的荧光化合物及其制备方法和应用 | |
| Wu et al. | Hydrogen peroxide-responsive AIE probe for imaging-guided organelle targeting and photodynamic cancer cell ablation | |
| CN111875603B (zh) | 一种β-咔啉吡啶鎓盐荧光探针及其制备方法与应用 | |
| CN106866721B (zh) | 一种硅酞菁衍生物及其制备生物素受体靶向硅酞菁光敏剂的应用 | |
| Mahajan et al. | Synthesis, photophysical properties and application of new porphyrin derivatives for use in photodynamic therapy and cell imaging | |
| Yang et al. | Diketopyrrolopyrrole-based multifunctional ratiometric fluorescent probe and γ-glutamyltranspeptidase-triggered activatable photosensitizer for tumor therapy | |
| CN110256313A (zh) | 一种光敏剂前药化合物及其制备方法和应用 | |
| CN114181253B (zh) | 一种基于氟硼二吡咯的线粒体靶向型光敏剂的制备及应用 | |
| Li et al. | Construction of a near-infrared fluorescent probe for ratiometric imaging of peroxynitrite during tumor progression | |
| CN111592482A (zh) | 一种pH可逆激活型光热/光动力/荧光一体化探针分子 | |
| Karges et al. | Synthesis, Characterisation and Biological Evaluation of π-Extended Fe (II) Bipyridine Complexes as Potential Photosensitizers for Photodynamic Therapy | |
| CN110857310B (zh) | 一种具有光活性的多联吡啶钌配合物及其应用 | |
| Krasnovskaya et al. | Photoinduced Reduction of Novel Dual-Action Riboplatin Pt (IV) Prodrug | |
| CN108640902A (zh) | 一种识别纯水体系内二氧化硫的荧光探针及其应用 | |
| Dukh et al. | Impact of mono-and di-β-galactose moieties in in vitro/in vivo anticancer efficacy of pyropheophorbide-carbohydrate conjugates by photodynamic therapy | |
| EP3757111A1 (en) | Ruthenium (ii) complexes and conjugates thereof for use as photosensitizer agent in photodynamic therapy | |
| EP3792265A1 (en) | Oxazine compound and application thereof | |
| Wang et al. | Synthesis and evaluation of new fluorinated pyropheophorbide-a derivatives for photodynamic therapy | |
| CN114315829A (zh) | 一类具有硝基还原酶响应的β-咔啉苯并[c,d]吲哚鎓盐及其制备方法和应用 | |
| CN109678888B (zh) | 噁嗪类化合物及其用途 | |
| Jiang et al. | Novel selenium-containing photosensitizers for near-infrared fluorescence imaging-guided photodynamic therapy | |
| Bornhuetter et al. | Synthesis, Photophysics and PDT Evaluation of Mono‐, Di‐, Tri‐and Hexa‐PEG Chlorins for Pointsource Photodynamic Therapy |
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 |