CN111978247B - 一种双光子荧光探针及其制备方法和应用 - Google Patents
一种双光子荧光探针及其制备方法和应用 Download PDFInfo
- Publication number
- CN111978247B CN111978247B CN202010827965.3A CN202010827965A CN111978247B CN 111978247 B CN111978247 B CN 111978247B CN 202010827965 A CN202010827965 A CN 202010827965A CN 111978247 B CN111978247 B CN 111978247B
- Authority
- CN
- China
- Prior art keywords
- photon
- fluorescent probe
- preparation
- formula
- photon fluorescent
- 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
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 45
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 42
- 150000001875 compounds Chemical class 0.000 claims description 24
- 239000000243 solution Substances 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000001514 detection method Methods 0.000 claims description 3
- 150000008044 alkali metal hydroxides Chemical class 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 238000000799 fluorescence microscopy Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 claims 1
- 230000005693 optoelectronics Effects 0.000 claims 1
- 239000007787 solid Substances 0.000 abstract description 20
- 238000003384 imaging method Methods 0.000 abstract description 16
- 230000015572 biosynthetic process Effects 0.000 abstract description 14
- 238000003786 synthesis reaction Methods 0.000 abstract description 14
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 abstract description 10
- 210000004027 cell Anatomy 0.000 abstract description 9
- 238000004020 luminiscence type Methods 0.000 abstract description 5
- 238000013461 design Methods 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 238000002428 photodynamic therapy Methods 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000002904 solvent Substances 0.000 abstract description 2
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 210000004881 tumor cell Anatomy 0.000 abstract description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 15
- 238000005160 1H NMR spectroscopy Methods 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 239000001257 hydrogen Substances 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- 238000004611 spectroscopical analysis Methods 0.000 description 6
- 238000000967 suction filtration Methods 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 5
- 239000008346 aqueous phase Substances 0.000 description 5
- 229940125904 compound 1 Drugs 0.000 description 5
- 229940125782 compound 2 Drugs 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 238000000295 emission spectrum Methods 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 230000002490 cerebral effect Effects 0.000 description 3
- 229940126214 compound 3 Drugs 0.000 description 3
- 229940125898 compound 5 Drugs 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 230000002792 vascular Effects 0.000 description 3
- AXPBAQGPJYCXKE-UHFFFAOYSA-N 1-[4-(ethylamino)phenyl]ethanone Chemical compound CCNC1=CC=C(C(C)=O)C=C1 AXPBAQGPJYCXKE-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 125000000609 carbazolyl group Chemical group C1(=CC=CC=2C3=CC=CC=C3NC12)* 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 230000000171 quenching effect Effects 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- DGPBVJWCIDNDPN-UHFFFAOYSA-N 2-(dimethylamino)benzaldehyde Chemical compound CN(C)C1=CC=CC=C1C=O DGPBVJWCIDNDPN-UHFFFAOYSA-N 0.000 description 1
- BGNGWHSBYQYVRX-UHFFFAOYSA-N 4-(dimethylamino)benzaldehyde Chemical compound CN(C)C1=CC=C(C=O)C=C1 BGNGWHSBYQYVRX-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 125000003342 alkenyl group Chemical group 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000010226 confocal imaging Methods 0.000 description 1
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- 125000001511 cyclopentyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100001085 no phototoxicity Toxicity 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D215/14—Radicals substituted by oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/06—Peri-condensed systems
-
- 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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- 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
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明属于有机荧光材料技术领域,具体涉及一种双光子荧光探针及其制备方法和应用。所述双光子荧光探针分子不含咔唑或其它较大的共轭结构,可以在固体条件下或者水相中表现出强的发光,该探针可以用于单光子和双光子细胞成像、小鼠脑血管双光子成像、肿瘤细胞的光动力治疗,具有很好的应用前景。并且其制备方法的设计、合成具有理论的指导,具有原料来源丰富、制备方法简单、不需要有毒溶剂、合成条件温和、合成产率高等优点,本发明制备得到的目标产品在醇的水溶液中为固体,通过抽滤即可分离,分离方法简单,产率高达90%以上。
Description
技术领域
本发明属于有机荧光材料技术领域,具体涉及一种双光子荧光探针及其制备方法和应用。
背景技术
荧光探针作为研究生物系统必不可少的工具,借助双光子显微成像可以直观便捷地对生物活性化合物或生物功能客体成分进行实时动态三维观测与监控。近十年来发展的双光子激发荧光探针,较单光子荧光探针具有显著的优点,如高分辨率、高清晰度、高灵敏度、无光漂白、无光致毒、定靶激发、高横向与纵向分辨率、低的生物组织吸光系数及低的组织自发荧光干扰等。双光子荧光探针已被广泛应用于临床诊断、疾病监测和药物筛选上,这推动了生物化学、医学和生命研究的发展。
例如,中国专利文献CN110003173A,公开了一种基于咔唑的双光子极性荧光探针及其制备方法和用途。该专利文献通过分子设计得到一种合适的荧光探针结构,使其具有基于咔唑的较大的共轭体系和共平面结构,双光子吸收性能优异,高选择性和透膜性,低细胞毒性,高光稳定性和生物相容性等优点,以实现定量检测溶液的极性变化和双光子共聚焦成像检测细胞凋亡过程中溶酶体的极性变化。但是,该基于咔唑基的双光子极性探针在有机溶液中具有很高的荧光量子效率,由于其在水相中或固体状态下表现出聚集淬灭的现象,严重限制了它们在生物检测、成像和光电器件中的应用前景。第二,该探针以咔唑为母体,其在合成过程中,产物的分离操作复杂,分离难度大,目标产物收率较低。
因此,开发在固体状态下或水相中具有强的发光的材料且合成步骤简单、收率高的新型双光子荧光探针将具有很好的应用前景。
发明内容
因此,本发明要解决的技术问题在于克服现有技术中的双光子荧光探针在水相中或固体状态下表现出聚集淬灭的现象,合成方法复杂、分离难度大、目标产品收率低等缺陷,从而提供一种双光子荧光探针及其制备方法和应用。
为此,本发明采用如下技术方案:
本发明提供一种双光子荧光探针,具有如下式所示结构:
其中,Ar1、Ar2独立的选自:
所述R、R1独立的选自具有1-16个碳原子的直链或支链烃基,具有6-16个碳原子的芳香环取代基中的一种。
进一步地,所述Ar1、Ar2独立的选自:
进一步地,所述R、R1独立的选自具有1-16个碳原子的直链或支链烷基、烯基、炔基,具有6-16个碳原子的芳香环取代基中的一种。
进一步地,所述R、R1独立的选自甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、环戊基、环己基、苯基或苄基中的一种;
优选的,所述所述R、R1独立的选自甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基中的一种。
进一步地,具有如下式之一所示的结构:
本发明还提供一种上述双光子荧光探针的制备方法,合成路线如下:
进一步地,式(I)所示化合物与式(II)所示化合物的摩尔比为1:0.9-1.1。
进一步地,所述碱金属氢氧化物为氢氧化钠或氢氧化钾;
所述醇溶液为醇的水溶液;优选的,所述醇溶液中醇和水的体积比为4-10:1;
优选的,所述醇为甲醇或乙醇。
进一步地,式(I)所示化合物与式(II)所示化合物的在0-80℃下反应,反应时间为45-50h,固液分离,得式(III)所示化合物;
优选的,在室温下反应。
本发明还提供一种上述双光子荧光探针在生物检测,荧光成像或光电器件中的应用。
本发明技术方案,具有如下优点:
1.本发明提供的新型双光子荧光探针,不含咔唑或其它较大的共轭结构,可以在固体条件下或者水相中表现出强的发光,该探针可以用于单光子和双光子细胞成像、小鼠脑血管双光子成像、肿瘤细胞的光动力治疗,具有很好的应用前景。
2.本发明提供的新型双光子荧光探针的制备方法,设计、合成具有理论的指导,并且其结构容易修饰(比如:引入PDT、细胞器或肿瘤靶向官能团等)、原料来源丰富、制备方法简单、不需要有毒溶剂、合成条件温和、合成产率高等优点,本发明制备得到的目标产品在醇的水溶液中为固体,通过抽滤即可分离,分离方法简单,产率高达90%以上,这为将来设计、合成近红外生物探针和光电器件具有重要的指导意义。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案,下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图是本发明的一些实施方式,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本发明实施例1的1H-NMR图;
图2为本发明实施例2的1H-NMR图;
图3为本发明实施例1和2固体状态在365nm紫外光照射下的图片;
图4为本发明实施例1在水相中的吸收和发射光谱对比;
图5为本发明实施例2在水相中的吸收和发射光谱对比;
图6为本发明实施例1双光子和单光子细胞成像对比;
图7为本发明实施例2双光子和单光子细胞成像对比;
图8为本发明实施例1和2小鼠脑血管成像对比。
具体实施方式
提供下述实施例是为了更好地进一步理解本发明,并不局限于所述最佳实施方式,不对本发明的内容和保护范围构成限制,任何人在本发明的启示下或是将本发明与其他现有技术的特征进行组合而得出的任何与本发明相同或相近似的产品,均落在本发明的保护范围之内。
实施例中未注明具体实验步骤或条件者,按照本领域内的文献所描述的常规实验步骤的操作或条件即可进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规试剂产品。
实施例1
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在1.91g(约10mmol)对乙胺基苯乙酮、1.89g(约10mmol)1-1、20mL的乙醇混合溶液(醇水体积比为7:1)中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.30g的化合物1(产率约91%)。
化合物1的1H NMR(400MHz,CDCl3)δ(ppm)=7.98(d,J=7.6Hz,2H),7.72(d,J=15.6Hz,1H),7.38–7.26(m,3H),6.65(d,J=7.6Hz,2H),6.56(d,J=8.4Hz,1H),3.44–3.34(m,6H),3.31(t,J=4.4Hz,2H),2.76(t,J=6.4Hz,2H),1.94(t,J=5.2Hz,2H),1.22–1.13(m,9H),通过核磁共振氢谱(图1)证明了化合物1的正确性。
实施例2
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在1.91g(约10mmol)对乙胺基苯乙酮、2.01g(约10mmol)1-1、20mL的乙醇混合溶液(醇水体积比为9:1中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.48g的化合物2(产率约93%)。
化合物2的1H NMR(400MHz,CDCl3)δ(ppm)=7.99(d,J=6.8Hz,2H),7.68(d,J=15.2Hz,1H),7.33(d,J=15.2Hz,1H),7.10(s,2H),6.66(d,J=8.0Hz,2H),3.43(q,J=5.2Hz,4H),3.23(t,J=5.1Hz,4H),2.76(t,J=6.1Hz,4H),2.00–1.93(m,4H),1.21(t,J=6.4Hz,6H),通过核磁共振氢谱(图2)证明了化合物2的正确性。
实施例3
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在1.49g(约10mmol)对二甲胺基苯甲醛、2.03g(约10mmol)3-1、20mL的乙醇混合溶液(醇水体积比为8:1中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.04g的化合物3(产率约91%)。
化合物3的1H NMR(400MHz,DMSO)δ(ppm)=7.63–7.53(m,5H),7.13(d,J=15.6Hz,1H),6.92(d,J=14.7Hz,1H),6.78(d,J=9.6Hz,2H),3.44–3.34(m,4H),3.02(s,6H)2.80(t,J=3.3Hz,2H),2.03–1.93(m,2H),1.12(t,J=3.3Hz,3H),通过核磁共振氢谱证明了化合物3的正确性。
实施例4
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在2.15g(约10mmol)4-0、2.03g(约10mmol)4-1、20mL的乙醇混合溶液(醇水体积比为8:1中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.61g的化合物4(产率约94%)。
化合物4的1H NMR(400MHz,CDCl3)δ(ppm)=7.59–7.53(m,2H),7.43(s,2H),7.13(d,J=9.3Hz,1H),7.02(s,1H),6.68(d,J=9.6Hz,1H),3.40–3.37(m,8H),2.79(t,J=5.4Hz,6H),1.99–1.93(m,6H),1.13(t,J=6.3Hz,3H),通过核磁共振氢谱证明了化合物4的正确性。
实施例5
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在2.15g(约10mmol)对5-0、2.01g(约10mmol)5-1、20mL的乙醇混合溶液(醇水体积比为10:1中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.59g的化合物5(产率约90%)。
化合物5的1H NMR(400MHz,CDCl3)δ(ppm)=7.59(d,J=6.6Hz,1H),7.43(s,2H),7.13(d,J=6.6Hz,1H),6.92(s,2H),3.37(t,J=5.4Hz,8H),2.79(t,J=6.3Hz,8H),2.00–1.93(m,8H),通过核磁共振氢谱证明了化合物5的正确性。
实施例6
本实施例提供一种新型双光子荧光探针的合成,合成路线如下:
在1.19g(约10mmol)对二甲氨基苯甲醛、2.15g(约10mmol)6-1、20mL的乙醇混合溶液(醇水体积比为5:1中,缓慢加入5mL氢氧化钠溶液(含1.00g氢氧化钠)。常温反应48h后,抽滤。得到橙红色固体,冷乙醇洗涤、干燥,得到3.15g的化合物6(产率约91%)。
化合物6的1H NMR(400MHz,DMSO)δ(ppm)=7.63–7.59(m,3H),7.44(s,2H),7.14(d,J=12.3Hz,1H),6.77(d,J=6.3Hz,2H),3.38(t,J=5.4Hz,4H),3.03(s,6H),2.80(t,J=6.3Hz,4H),2.01–1.95(m,4H),通过核磁共振氢谱证明了化合物6的正确性。
实验例
1、发光性能测试
(1)将本发明实施例1和2制备得到的化合物装入试管中,在365nm紫外光照射条件下,固体化合物1和2的发光图片入图3所示(图中,数字1代表化合物1,数字2代表化合物2),图3证明了材料可以在固体条件下发光;
(2)采用如下方法分别检测化合物1、化合物2的吸收光谱:
分别用分光光度计和发光光谱仪测试化合物在2mL DMSO溶液(10-6M)中的吸收和发射光谱。通过吸收和发射光谱(图4和图5)对比,证明了化合物1和2在水相中具有强的发射荧光。
2、细胞成像实验
将染料溶于DMSO溶液中,配制成10-5mol/L的溶液,将2μL上述溶液加入2mL细胞培养介质中,由10%胎牛血清以及90%含青霉素(80U/mL)和链霉素(80μg/mL)的不完全DMEM(高糖)培养基所配成的DMEM培养液,以此作为培养基在37℃下培养SKOV3细胞1小时,通过激光共聚焦扫描显微镜,分别以405nm和808nm作为激发波长,可以在红光通道下观察到在细胞中有较强的信号。通过化合物1和2的细胞成像实验(图6和图7),证明了它们可以用于单光子和双光子激发的细胞成像。
3、小鼠脑血管成像实验
将染料溶于DMSO溶液中,配制成10-3mol/L的溶液,将30μL上述溶液加入150μLPBS(磷酸缓冲溶液)中,稀释成2×10-4mol/L的溶液。通过眼球注射,将稀释后的溶液注射到小鼠体内,1h后监测小鼠脑血管成像。
通过化合物1和2的小鼠脑血管成像实验(图8),证明了它们可以用于小鼠脑血管成像。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引申出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (8)
3.根据权利要求2所述的双光子荧光探针的制备方法,其特征在于,式I所示化合物与式1-1或式2-1所示化合物的摩尔比为1:0.9-1.1。
4.根据权利要求2或3所述的双光子荧光探针的制备方法,其特征在于,所述碱金属氢氧化物为氢氧化钠或氢氧化钾;
所述醇溶液为醇的水溶液;所述醇溶液中醇和水的体积比为4-10:1。
5.根据权利要求4所述的双光子荧光探针的制备方法,其特征在于,所述醇为甲醇或乙醇。
6.根据权利要求2所述的双光子荧光探针的制备方法,其特征在于,式I所示化合物与式1-1或式2-1所示化合物在0-80℃下反应,反应时间为45-50h,固液分离,得式(III)所示化合物。
7.根据权利要求6所述的双光子荧光探针的制备方法,其特征在于,在室温下反应。
8.一种权利要求1所述的双光子荧光探针或权利要求2-7任一项所述制备方法制备得到的双光子荧光探针在生物检测,荧光成像或光电器件中非疾病诊断或治疗目的的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010827965.3A CN111978247B (zh) | 2020-08-17 | 2020-08-17 | 一种双光子荧光探针及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010827965.3A CN111978247B (zh) | 2020-08-17 | 2020-08-17 | 一种双光子荧光探针及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111978247A CN111978247A (zh) | 2020-11-24 |
CN111978247B true CN111978247B (zh) | 2022-01-25 |
Family
ID=73434586
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010827965.3A Active CN111978247B (zh) | 2020-08-17 | 2020-08-17 | 一种双光子荧光探针及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111978247B (zh) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1143340A (en) * | 1967-02-03 | 1969-02-19 | Eastman Kodak Co | Electrophotographic material |
JPS63145302A (ja) * | 1986-12-09 | 1988-06-17 | Canon Inc | 光重合開始剤及び記録媒体 |
JPH10300952A (ja) * | 1997-04-25 | 1998-11-13 | Tomoegawa Paper Co Ltd | 光導波回路及びその位相調整方法 |
CN103097339A (zh) * | 2010-08-27 | 2013-05-08 | 新加坡国立大学 | 用于胚胎干细胞探测的查耳酮结构荧光染料 |
CN108795088A (zh) * | 2018-04-17 | 2018-11-13 | 南京邮电大学 | 一种具有增强光动力和光热效果的近红外染料及其制备和应用 |
CN110003173A (zh) * | 2019-04-26 | 2019-07-12 | 安徽大学 | 一种基于咔唑的双光子极性荧光探针及其制备方法和用途 |
-
2020
- 2020-08-17 CN CN202010827965.3A patent/CN111978247B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1143340A (en) * | 1967-02-03 | 1969-02-19 | Eastman Kodak Co | Electrophotographic material |
JPS63145302A (ja) * | 1986-12-09 | 1988-06-17 | Canon Inc | 光重合開始剤及び記録媒体 |
JPH10300952A (ja) * | 1997-04-25 | 1998-11-13 | Tomoegawa Paper Co Ltd | 光導波回路及びその位相調整方法 |
CN103097339A (zh) * | 2010-08-27 | 2013-05-08 | 新加坡国立大学 | 用于胚胎干细胞探测的查耳酮结构荧光染料 |
CN108795088A (zh) * | 2018-04-17 | 2018-11-13 | 南京邮电大学 | 一种具有增强光动力和光热效果的近红外染料及其制备和应用 |
CN110003173A (zh) * | 2019-04-26 | 2019-07-12 | 安徽大学 | 一种基于咔唑的双光子极性荧光探针及其制备方法和用途 |
Non-Patent Citations (3)
Title |
---|
Characterization of the Fluorescence Properties of 4-Dialkylaminochalcones and Investigation of the Cytotoxic Mechanism of Chalcones;Bo Zhou et al.;《Arch. Pharm. Chem. Life Sci. 》;20160523;第349卷;第1-14页 * |
Characterization of the Fluorescence Properties of 4-Dialkylaminochalcones and Investigation of the Cytotoxic Mechanism of Chalcones;Bo Zhou et al.;《Arch. Pharm. Chem. Life Sci.》;20160523;第349卷;第1-14页 * |
Fluorescence water sensor based on covalent immobilization of chalcone derivative;Cheng-Gang Niu et al.;《Analytica Chimica Acta》;20060627;第577卷;第264-270页 * |
Also Published As
Publication number | Publication date |
---|---|
CN111978247A (zh) | 2020-11-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Jiang et al. | A NIR BODIPY dye bearing 3, 4, 4 a-trihydroxanthene moieties | |
CN102702768B (zh) | 一种新型红光bodipy荧光染料及其制备方法和应用 | |
CN111995579B (zh) | 一种含咪唑环结构的四苯乙烯衍生物制备方法和应用 | |
Liu et al. | A series of carbazole cationic compounds with large two-photon absorption cross sections for imaging mitochondria in living cells with two-photon fluorescence microscopy | |
CN110305026B (zh) | 固体荧光染料及其制备方法 | |
CN113620963B (zh) | 一种线粒体粘度探针及其制备方法和应用 | |
Han et al. | A diphenylacrylonitrile conjugated porphyrin with near-infrared emission by AIE–FRET | |
CN111333660B (zh) | 一类550nm激发的罗丹明类染料及其制备方法 | |
CN109369684B (zh) | 一类电子供体-受体-供体荧光分子及制备方法和应用 | |
CN111793371B (zh) | 一种3,5位不对称修饰bodipy类近红外荧光染料及其制备方法 | |
CN111978247B (zh) | 一种双光子荧光探针及其制备方法和应用 | |
US11639360B2 (en) | Oxazine compound and application thereof | |
CN111961072B (zh) | 一种溶酶体靶向的红外二窗发射荧光染料及其制备方法和应用 | |
CN115385825A (zh) | 一种具有活性氧产生能力的聚集诱导发光特性光敏剂及其制备方法和应用 | |
CN113004264B (zh) | 一种聚集诱导发光光敏剂、制备方法和应用 | |
CN114907311A (zh) | 一种基于aie性能的脂滴特异性荧光探针及制备方法和应用 | |
CN112552901B (zh) | 一种比率型锌离子荧光探针及其制备与应用 | |
CN111334080B (zh) | 一种高亮度、高光稳定性的碳酸酐酶荧光探针 | |
Guo et al. | One-pot synthesis and applications of two asymmetrical benzoxanthene dyes | |
Li et al. | Photostable fluorescent probes based on multifunctional group substituted naphthalimide dyes for imaging of lipid droplets in live cells | |
CN116836565B (zh) | 一类水溶性方酸菁染料及其合成方法和应用 | |
CN114315643B (zh) | 一种靶向脂滴和水环境的双色荧光探针及其合成方法和应用 | |
CN115353460B (zh) | 一种含苯酚的酮基-水杨醛联肼类化合物在内质网成像中的应用 | |
CN115960087B (zh) | 黏度响应型双光子荧光化合物及其合成与应用 | |
KR100870243B1 (ko) | 세포의 실시간 모니터링용 이광자 염료, 그 제조방법 및이를 이용한 세포의 실시간 모니터링 방법 |
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 |