WO2016133218A1 - Composé de phospha-fluorescéine ou son sel, et colorant fluorescent l'utilisant - Google Patents

Composé de phospha-fluorescéine ou son sel, et colorant fluorescent l'utilisant Download PDF

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WO2016133218A1
WO2016133218A1 PCT/JP2016/054964 JP2016054964W WO2016133218A1 WO 2016133218 A1 WO2016133218 A1 WO 2016133218A1 JP 2016054964 W JP2016054964 W JP 2016054964W WO 2016133218 A1 WO2016133218 A1 WO 2016133218A1
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group
compound
salt
phosphafluorescein
general formula
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PCT/JP2016/054964
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English (en)
Japanese (ja)
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山口　茂弘
愛子 中
真司 須田
正泰 多喜
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国立大学法人名古屋大学
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    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B11/00—Diaryl- or thriarylmethane dyes
    • C09B11/28—Pyronines ; Xanthon, thioxanthon, selenoxanthan, telluroxanthon dyes
    • 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
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers

Definitions

  • the present invention relates to a phosphafluorescein compound or a salt thereof, or a fluorescent dye using the same.
  • a dye having absorption and fluorescence maximum wavelengths in this region (620 nm or more) has attracted attention in deep observation of living tissue.
  • the fluorescence quantum yield is increased while having a fluorescence maximum wavelength at a position of about 600 to 700 nm. It is preferable to improve.
  • fluorescent dyes exhibiting water solubility there are not so many dyes having a fluorescence maximum at 650 nm or more compared to green or yellow phosphors (dyes having a fluorescence maximum at 495 to 570 nm).
  • a fluorescein dye has an absorption maximum wavelength of 491 nm, a fluorescence maximum wavelength of 510 nm, and a quantum yield of 0.85 (Non-Patent Document 1). Since it is necessary to exhibit absorption and fluorescence with respect to the wavelength, there is a demand for a dye having an absorption maximum and a fluorescence maximum at a wavelength of about 600 to 700 nm.
  • Non-Patent Document 2 it is known that when the oxygen atom of the xanthene skeleton of this fluorescein dye is replaced with a silicon atom, the absorption maximum wavelength is 582 nm, the fluorescence maximum wavelength is 598 nm, and the quantum yield is 0.42, which can be shifted to the longer wavelength side.
  • Non-Patent Document 2 the effect is insufficient and further long wavelength shifts are necessary.
  • a dye having an absorption maximum and a fluorescence maximum in a long wavelength region has a low quantum yield and is difficult to sufficiently increase.
  • these fluorescent dyes have a high maximum occupied orbit (HOMO), and thus react with oxygen under light irradiation to give active oxygen species. Therefore, the stability to light is low.
  • HOMO maximum occupied orbit
  • the present invention provides a compound (fluorescent dye) capable of obtaining a sufficient quantum yield and sufficiently reducing HOMO while having an absorption maximum wavelength and a fluorescence maximum wavelength in the range of 600 to 700 nm. ).
  • the present inventors have developed a series of red fluorescent dyes (phosphafluorescein compounds) in which the oxygen atom at the 10-position of the xanthene ring portion of fluorescein is substituted with a phosphorus atom.
  • This compound is anionized under neutral or alkaline conditions, and has a sufficiently high fluorescence quantum yield even under physiological conditions while having an absorption maximum wavelength and a fluorescence maximum wavelength of 600 to 700 nm. We found that rate was obtained.
  • this compound lowered the HOMO level due to the electronic effect of the phosphorus substituent, a remarkable stabilizing effect against light irradiation was also observed.
  • the present invention has been completed by further research based on such knowledge. That is, the present invention includes the following configurations.
  • R 1 represents an optionally substituted aryl group.
  • R 2 represents a hydrogen atom or an organic group.
  • R is the general formula (1A) to (1C):
  • R 3 represents an optionally substituted aryl group, an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group.
  • R 4 represents a substituted group. Represents an alkyl group which may be substituted.) It is group represented by these. ] Or a salt, hydrate or solvate thereof.
  • Item 2 The phosphafluorescein compound according to item 1, or a salt, hydrate or solvate thereof, wherein in the general formula (1), R is a group represented by the general formula (1A).
  • Item 3 The phosphafluorescein compound according to Item 1 or 2, or a salt, hydrate, or solvate thereof, having an anion represented by:
  • Item 4 The phosphafluorescein compound or a salt, hydrate or solvate thereof according to any one of Items 1 to 3, which has a maximum absorption wavelength at 600 to 700 nm.
  • Item 5 The phosphafluorescein compound according to any one of Items 1 to 4, having a maximum absorption wavelength at 600 to 700 nm and a fluorescence quantum yield of 0.25 to 0.60, or a salt, hydrate or Solvate.
  • Item 6. A fluorescent dye containing the phosphafluorescein compound according to any one of Items 1 to 5 or a salt, hydrate or solvate thereof.
  • Item 7. The fluorescent dye according to Item 6, which is a fluorescent dye for bioimaging.
  • Item 8 The fluorescent dye according to Item 7, which is a fluorescent dye for cancer cell bioimaging.
  • Item 9. A cell detection agent comprising the fluorescent dye according to any one of Items 6 to 8.
  • Item 10 The cell detection agent according to Item 9, which is a cancer cell detection agent.
  • Item 11 A cell bioimaging method using the fluorescent dye according to any one of Items 6 to 8, or the cell detection agent according to Item 9 or 10.
  • Item 12. The bioimaging method according to Item 11, wherein the cell is a cancer cell.
  • the phosphafluorescein compound of the present invention or a salt, hydrate or solvate thereof has an absorption maximum wavelength and a fluorescence maximum wavelength of 600 to 700 nm because the oxygen atom at the 10-position of the xanthene ring portion of fluorescein is substituted with a phosphorus atom.
  • a sufficiently high fluorescence quantum yield can be obtained even under physiological conditions.
  • the absorption maximum wavelength of 627 nm in the examples is almost the same as the excitation wavelength of 633 nm of the HeNe laser normally provided in a laser microscope, so that the excitation efficiency as a fluorescent dye is extremely high.
  • the phosphafluorescein compound of the present invention or a salt thereof can reduce the HOMO level due to the electronic effect of the phosphorus substituent, and can dramatically improve the stability to light. It is possible to observe for a long time.
  • FIG. 3 is a result of X-ray crystal structure analysis of a phosphafluorescein compound (compound POF) obtained in Example 1.
  • FIG. 2 is an absorption spectrum and a fluorescence spectrum of the phosphafluorescein compound (compound POF) of the present invention when adjusted to pH 3, pH 7, and pH 9 prepared in Test Example 3.
  • FIG. It is an absorption spectrum (upper figure) of the phosphafluorocein compound (compound POF) of the present invention at each pH, and a graph (lower figure) showing the relationship between relative absorption at 627 nm and pH.
  • (A) is a graph showing the absorbance maintenance rate at the absorption maximum wavelength when the phosphafluorescein compound of the present invention and known fluorescein compounds (TokyoGreen and TokyoMagenta) are used.
  • (B) is a graph showing the absorbance maintenance rate at 630 nm of test solutions 7 to 10 prepared in Test Example 5.
  • FIG. 6 is a photograph showing the results of cell incubation and imaging in Test Example 6.
  • the left figure (a) is a Confocal fluorescence image obtained by excitation at 633 nm.
  • the middle figure (b) is a bright field image.
  • the right figure (c) is a merged image of (a) and (b). In the figure, the scale bar is 50 ⁇ m.
  • 6 is a graph showing the results of cytotoxicity in Test Example 6.
  • Phosphafluorescein compound or a salt, hydrate or solvate thereof has the general formula (1):
  • R 1 represents an optionally substituted aryl group.
  • R 2 represents a hydrogen atom or an organic group.
  • R is the general formula (1A) to (1C):
  • R 3 represents an optionally substituted aryl group, an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group.
  • R 4 represents a substituted group. Represents an alkyl group which may be substituted.) It is group represented by these. ] Or a salt thereof.
  • the phosphafluorescein compound represented by the general formula (1) or a salt thereof is a novel compound not described in any literature.
  • any of a monocyclic aryl group, a polycyclic aryl group, and a heteroaromatic ring group can be employed.
  • the substituent which the aryl group represented by R 1 may have is not particularly limited, and is a hydroxyl group, a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.), alkyl group (methyl group, ethyl group). Group, n-propyl group, etc.), formyl group, carboxyl group, ester group, amide group (amide group, methylamide group, dimethylamide group etc.), azido group (azidephenyl group etc.), alkynyl group (ethynyl group, propargyl group) Etc.), alkenyl groups (vinyl group, allyl group, etc.) and the like.
  • the number of such substituents is not particularly limited and is preferably 0 to 6, more preferably 0 to 3.
  • R 1 is preferably a substituted or unsubstituted monocyclic aryl group (substituted or unsubstituted phenyl group) from the viewpoint of further improving the fluorescence quantum yield and making it easier to recognize during bioimaging.
  • a substituted monocyclic aryl group having a substituent at the position (substituted phenyl group) is more preferred, and an o-tolyl group is more preferred.
  • the organic group represented by R 2 may be an alkyl group which may have a substituent, an acyl group which may have a substituent, or a substituent.
  • Good cyclic ether groups and the like can be mentioned.
  • alkyl group as the organic group represented by R 2 , both a linear alkyl group and a branched alkyl group can be employed.
  • a straight-chain alkyl group having 1 to 6 carbon atoms is preferable.
  • Specific examples include a methyl group, an ethyl group, an n-propyl group, an n-butyl group, and an n-pentyl group.
  • branched alkyl group a branched alkyl group having 3 to 6 carbon atoms (particularly 3 to 5 carbon atoms) is preferable.
  • isopropyl group, isobutyl group, t-butyl group, s-butyl group, neopentyl group examples thereof include an isohexyl group and a 3-methylpentyl group.
  • the substituent that the alkyl group as the organic group represented by R 2 may have is not particularly limited, and examples thereof include a hydroxyl group and a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.). .
  • the number of such substituents is not particularly limited and is preferably 0 to 6, more preferably 0 to 3.
  • examples of the acyl group as the organic group represented by R 2 include a methanoyl group and an ethanoyl group.
  • examples of the cyclic ether group as the organic group represented by R 2 include a tetrahydropyranyl group and a tetrahydrofuranyl group.
  • the substituent that the cyclic ether as the organic group represented by R 2 may have is not particularly limited, and is a hydroxyl group, a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.), acyl group ( Methanoyl group, ethanoyl group, etc.), silyl group (trimethylsilyl group, triethylsilyl group, etc.) and the like.
  • the number of such substituents is not particularly limited, but is preferably 0 to 2, more preferably 0 to 1.
  • R 2 is preferably a hydrogen atom, an acyl group, or a substituted cyclic ether group, and more preferably a hydrogen atom, from the viewpoint of easy anionization and easy shift of the absorption maximum wavelength and the fluorescence maximum wavelength.
  • both a monocyclic aryl group and a polycyclic aryl group are employed.
  • examples thereof include a phenyl group, an oligoaryl group (naphthyl group, anthryl group, etc.), a biphenyl group, a terphenyl group, a pyrenyl group, a phenanthrenyl group, and a fluorenyl group.
  • the substituent which the aryl group represented by R 3 may have is not particularly limited, and is a hydroxyl group, a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.), alkyl group (methyl group, ethyl group). Group, n-propyl group, etc.).
  • the number of such substituents is not particularly limited, but is preferably 0 to 6, more preferably 0 to 3.
  • a straight-chain alkyl group having 1 to 6 carbon atoms is preferable.
  • Specific examples include a methyl group, an ethyl group, an n-propyl group, an n-butyl group, and an n-pentyl group.
  • branched alkyl group a branched alkyl group having 3 to 6 carbon atoms (particularly 3 to 5 carbon atoms) is preferable.
  • isopropyl group, isobutyl group, t-butyl group, s-butyl group, neopentyl group examples thereof include an isohexyl group and a 3-methylpentyl group.
  • the substituent that the alkyl group as the organic group represented by R 2 may have is not particularly limited, and examples thereof include a hydroxyl group and a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.). .
  • the number of such substituents is not particularly limited, but is preferably 0 to 6, more preferably 0 to 3.
  • examples of the alkenyl group represented by R 3 include a vinyl group and an allyl group.
  • examples of the alkynyl group represented by R 3 include an alkynyl group and a propargyl group.
  • the electronic structure can be finely adjusted and the physical properties (solubility, cell permeability, etc.) can be adjusted.
  • a straight-chain alkyl group having 1 to 6 carbon atoms is preferable.
  • Specific examples include a methyl group, an ethyl group, an n-propyl group, an n-butyl group, and an n-pentyl group.
  • branched alkyl group a branched alkyl group having 3 to 6 carbon atoms (particularly 3 to 5 carbon atoms) is preferable.
  • isopropyl group, isobutyl group, t-butyl group, s-butyl group, neopentyl group examples thereof include an isohexyl group and a 3-methylpentyl group.
  • the substituent that the alkyl group as the organic group represented by R 2 may have is not particularly limited, and examples thereof include a hydroxyl group and a halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom, etc.). .
  • the number of such substituents is not particularly limited, but is preferably 0 to 6, more preferably 0 to 3.
  • R has higher electron withdrawing properties, and it is easier to reduce the HOMO level (energy level of the highest occupied orbit) and LUMO level (energy level of the lowest unoccupied orbit). From the viewpoint of easily improving the stability, a group represented by the general formula (1A) is preferable.
  • Examples of the compound of the present invention that satisfies such conditions include, for example,
  • Ph represents a phenyl group. The same applies hereinafter.
  • a phosphafluorescein compound represented by the above, or a salt, hydrate or solvate thereof is preferred.
  • the compound of the present invention having such a structure has a high electron withdrawing property due to the presence of a phosphorus-containing group, and the HOMO level (energy level of the highest occupied orbit) and LUMO level (energy level of the lowest unoccupied orbit). ) Can be reduced.
  • the LUMO level can be effectively reduced compared to the HOMO level, so the HOMO-LUMO energy gap when changed to an anion type is reduced to 2.00-2.50 eV, especially 2.25-2.48 eV. (B3LYP / 6-31 + G value obtained by quantum chemistry calculations performed at the G level). For this reason, it is also possible to shift the absorption maximum wavelength and the fluorescence maximum wavelength by longer wavelengths.
  • HOMO and LUMO levels are measured by structural optimization using Gaussian 09 program.
  • a neutral compound (compound represented by the general formula (1)) can have a fluorescence maximum wavelength of about 600 to 650 nm, particularly 610 to 640 nm.
  • the salt form of the compound represented by the general formula (1) is anionized, the absorption maximum wavelength is shifted to a wavelength longer than 600 to 700 nm (particularly about 600 to 650 nm). It is also possible to further shift the fluorescence maximum wavelength (about 650 to 700 nm, particularly about 655 to 670 nm) and further improve the fluorescence quantum yield (about 0.25 to 0.60, especially 0.35 to 0.50).
  • the compound of the present invention is preferably a salt of the compound represented by the general formula (1), and the general formula (2):
  • R 1 and R are the same as those described above.
  • the compound of the present invention may exist as a hydrate or a solvate, and any of these substances is included in the scope of the present invention.
  • the phosphafluorescein compound of the present invention or a salt, hydrate or solvate thereof is not particularly limited, and examples thereof include silicon-substituted fluorescein compounds. It can be synthesized according to the method already reported for the production method (Chem. Commun. 2011, 47, 4162.) (except for changing the raw material compound).
  • R and R 1 are the same as defined above.
  • R 5 and R 6 are the same or different and represent a protecting group.
  • X 1 and X 2 are the same or different and each represents a halogen atom.
  • the protecting group represented by R 5 and R 6 is not particularly limited as long as it is a group capable of protecting a hydroxyl group, and any group can be used.
  • an alkanoyl group (formyl group, C1-C4 alkanoyl groups such as acetyl group and propionyl), optionally substituted aralkyl groups (benzyl group, p-methoxybenzyl group, p-nitrobenzyl group, etc.), silyl groups (trimethylsilyl group, triethylsilyl group) Group, t-butyldimethylsilyl group, etc.), alkoxyalkyl group (methoxymethyl group etc.), tetrahydropyranyl (THP) group and the like.
  • a silyl group is preferable and a t-butyldimethylsilyl group is more preferable from the viewpoint of protecting the hydroxyl group and facilitating the progress of the nucleophilic addition
  • any of a fluorine atom, a chlorine atom, a bromine atom and an iodine atom can be adopted, but from the viewpoint of easy progress of the cyclization reaction, a chlorine atom , A bromine atom, an iodine atom and the like are preferable, and a bromine atom is more preferable.
  • the compound (3) in the above reaction formula 1 can be a known or commercially available compound, or can be synthesized. In the case of synthesis, it can be synthesized by reacting 3-halogenated-N, N-diallylaniline obtained by reacting 3-halogenated aniline and allyl halide with formaldehyde.
  • the organic lithium compound is not particularly limited, and known compounds can be used.
  • Alkyl lithium such as lithium
  • cycloalkyl lithium such as cyclohexyl lithium
  • aryl lithium such as phenyl lithium; and the like.
  • alkyllithium is preferable and n-butyllithium is more preferable from the viewpoint of yield.
  • the organophosphorus compound is not particularly limited as long as a compound having a group represented by R can be obtained in this step, and known compounds can be used, for example, dihalogenated arylphosphine such as dichlorophenylphosphine and dibromophenylphosphine. Can be used.
  • the amount of the organolithium compound, organophosphorus compound and hydrogen peroxide used is not particularly limited, and from the viewpoint of yield and the like, 1 to 20 moles (especially 2 to 2 moles) of the organolithium compound per mole of the compound (3) It is preferable to use 0.1 to 10 mol (especially 0.5 to 5 mol) of the organic phosphorus compound. Further, it is preferable to use an excess amount of an aqueous solution of hydrogen peroxide.
  • the organic solvent that can be used in this step known ones can be adopted, and in this step, for example, cyclic ethers such as tetrahydrofuran and dioxane are preferable.
  • the reaction conditions are preferably such that the reaction proceeds sufficiently.
  • the reaction can be carried out at ⁇ 150 to 0 ° C., particularly ⁇ 100 to ⁇ 50 ° C. for 5 minutes to 12 hours, particularly 10 minutes to 6 hours.
  • the oxidizing agent is not particularly limited, and permanganate (such as potassium permanganate) can be used.
  • the amount of the oxidizing agent used is not particularly limited, and is preferably used in an amount of 1 to 10 mol (especially 2 to 5 mol) with respect to 1 mol of compound (4) from the viewpoint of yield and the like.
  • the organic solvent that can be used in this step a known one may be employed.
  • cyclic ethers such as tetrahydrofuran and dioxane are preferable.
  • the same solvent as the said cyclization process can be used.
  • the reaction conditions may be such that the reaction proceeds sufficiently.
  • the reaction conditions may be -50 to 100 ° C., particularly 0 to 50 ° C., 1 to 48 hours, particularly 2 to 24 hours.
  • the amount of the palladium catalyst and 1,3-dimethylbarbituric acid used is not particularly limited. From the viewpoint of yield and the like, 0.1 to 1 mol of palladium catalyst (especially 0.2 to 1 mol) per 1 mol of compound (5). 0.5 mol) and 2 to 50 mol (especially 5 to 30 mol) of 1,3-dimethylbarbituric acid are preferably used.
  • the organic solvent that can be used in this step a known one may be employed.
  • cyclic ethers such as tetrahydrofuran and dioxane are preferable.
  • the same solvent as the said cyclization process can be used.
  • the reaction conditions may be such that the reaction proceeds sufficiently.
  • the reaction conditions may be 1 to 96 hours, particularly 2 to 72 hours at 0 to 150 ° C., particularly 50 to 100 ° C.
  • nitrous acid or a salt thereof in addition to nitrous acid, nitrite (sodium nitrite) or the like can be adopted.
  • the amount of nitrous acid or a salt thereof used is not particularly limited, but from the viewpoint of yield and the like, it is preferable to use 1 to 10 mol (particularly 2 to 5 mol) with respect to 1 mol of compound (6). .
  • the organic solvent that can be used in this step known ones can be adopted, and in this step, for example, cyclic ethers such as tetrahydrofuran and dioxane are preferable.
  • the same solvent as the said cyclization process can be used.
  • the reaction conditions may be such that the reaction proceeds sufficiently, for example, 50 to 200 ° C., particularly 100 to 150 ° C., 5 minutes to 5 hours, particularly 10 minutes to 2 hours.
  • R 1 MgX 3 organic magnesium represented by R 1 MgX 3 (wherein R 1 is the same as above, X 3 represents a halogen atom). Compounds are preferred.
  • halogen atom represented by X 3 those described above can be adopted. The same applies to preferred embodiments.
  • Grignard reagents that satisfy these conditions include:
  • the amount of Grignard reagent and oxidizing agent used is not particularly limited, and from the viewpoint of yield, etc., 1 to 10 moles (especially 2 to 5 moles) of Grignard reagent should be used per 1 mole of compound (5). Is preferred.
  • the oxidizing agent is preferably used in an excess amount as an aqueous solution.
  • the organic solvent that can be used in this step known ones can be adopted, and in this step, for example, cyclic ethers such as tetrahydrofuran and dioxane are preferable.
  • the same solvent as the said cyclization process can be used.
  • the reaction conditions may be such that the reaction proceeds sufficiently.
  • the reaction conditions may be ⁇ 50 to 100 ° C., particularly 0 to 50 ° C., 30 minutes to 10 hours, particularly 1 to 5 hours.
  • the phosphafluorescein compound represented by the general formula (1) can be obtained, and can be used through ordinary isolation and purification steps as necessary.
  • the phosphafluorescein compound represented by the general formula (1) is a neutral compound under acidic conditions (for example, pH 1 to 5), but is made to be under neutral conditions or alkaline conditions (for example, pH 6 to 14). Thus, it can be easily converted into a salt having an anion represented by the general formula (2), and can be used through ordinary isolation and purification steps as necessary.
  • the absorption maximum wavelength is shifted to a longer wavelength in the range of 600 to 700 nm (especially about 600 to 650 nm) and the fluorescence maximum wavelength is further shifted (about 650 to 700 nm, particularly 655 to 650 nm). It is possible to further improve the fluorescence quantum yield (about 0.25 to 0.60, especially 0.35 to 0.50).
  • the fluorescent dye of the present invention contains the phosphafluorescein compound of the present invention or a salt thereof.
  • the oxygen atom at the 10-position of the xanthene ring portion of fluorescein is substituted with a phosphorus atom, so that it has a maximum absorption wavelength and a maximum fluorescence wavelength in the range of 600 to 700 nm, but it is sufficient even under physiological conditions.
  • a high fluorescence quantum yield is obtained.
  • the absorption maximum wavelength of 627 nm in the examples is almost the same as the excitation wavelength of 633 nm of a HeNe laser that is normally equipped with a laser microscope, so that the excitation efficiency as a fluorescent dye is extremely high.
  • the phosphafluorescein compound or a salt thereof of the present invention has high permeability into cells, and when a desired substituent is introduced as R 1 , desired cells (for example, cancer cells such as HeLa cells) in vivo. ) Can be selectively localized. That is, only desired cells (for example, cancer cells such as HeLa cells) can emit light.
  • the phosphafluorescein compound or a salt thereof of the present invention has a high fluorescence quantum that is not found in conventional fluorescent dyes that have an absorption maximum wavelength and a fluorescence maximum wavelength in the range of 600 to 700 nm but have a maximum wavelength in this region.
  • the phosphafluorescein compound of the present invention or a salt thereof can reduce the HOMO level by the electronic effect of the phosphorus substituent, and can dramatically improve the stability to light. It is possible to observe for a long time.
  • the cell detection agent of the present invention contains the phosphafluorescein compound of the present invention or a salt thereof, preferably dissolved in an organic solvent to form a solution, From the viewpoint of detecting more desired cells (for example, cancer cells such as HeLa cells) and color rendering (visualization in real time) the desired cells (for example, cancer cells such as HeLa cells) more selectively.
  • the content of the fluorescein compound is preferably 1 ⁇ 10 ⁇ 8 to 1 ⁇ 10 ⁇ 4 mol / L, more preferably 1 ⁇ 10 ⁇ 7 to 1 ⁇ 10 ⁇ 5 mol / L.
  • content of a phosphafluorescein compound can be restrained low.
  • the organic solvent that can be used is not particularly limited, and both polar solvents and nonpolar solvents are used. it can.
  • polar solvents examples include ether compounds (tetrahydrofuran, anisole, 1,4-dioxane, cyclopentylmethyl ether, etc.), alcohols (methanol, ethanol, allyl alcohol, etc.), ester compounds (ethyl acetate, etc.), ketones (acetone, etc.) , Halogenated hydrocarbons (dichloromethane, chloroform), dimethyl sulfoxide, amide solvents (N, N-dimethylformamide, dimethylacetamide, 1,3-dimethyl-2-imidazolidinone, N-methylpyrrolidone, etc.) .
  • ether compounds tetrahydrofuran, anisole, 1,4-dioxane, cyclopentylmethyl ether, etc.
  • alcohols methanol, ethanol, allyl alcohol, etc.
  • ester compounds ethyl acetate, etc.
  • ketones acetone, etc.
  • Halogenated hydrocarbons
  • nonpolar solvent examples include aliphatic organic solvents such as pentane, hexane, cyclohexane and heptane; aromatic solvents such as benzene, toluene, xylene and mesitylene.
  • the cell detection agent of the present invention is preferably in the form of a solution, but has a sufficiently high fluorescence quantum yield even under physiological conditions while having an absorption maximum wavelength and a fluorescence maximum wavelength at 600 to 700 nm.
  • the pH is preferably about 5 to 11 and more preferably about 6.5 to 7.5 from the viewpoint of introduction into cells.
  • Buffers Hepes buffer, Tris buffer, tricine-sodium hydroxide buffer, phosphate buffer, phosphate buffered saline, etc. are used to adjust the pH of the cell detection agent of the present invention. May be.
  • the general operating melting point (mp) or decomposition temperature was measured with a Yanaco MP-S3 instrument (MP-S3).
  • 1 H, 13 C ⁇ 1 H ⁇ and 31 P ⁇ 1 H ⁇ NMR spectra were measured using JEOL AL-400 spectrometer (400 MHz for 1 H, 100 MHz for 13 C and 161.70 MHz for 31 P), JEOL JNM-ECS400 ( CDCl 3 , CD 2 Cl 2 or 400 MHz for 1 H, 100 MHz for 13 C and 161.70 MHz for 31 P) or JEOL A-600 spectrometer (600 MHz for 1 H and 150 MHz for 13 C) Measured in CD 3 OD.
  • Mass spectra were measured by electrospray ionization (ESI) using a Bruker micrOTOF Focus spectrometry system using atmospheric pressure chemical ionization (APCI) or Thermo Fisher Scientific Exactive.
  • Thin layer chromatography (TLC) was performed using a glass plate coated with silica gel 60F 254 (Merck).
  • Column chromatography was performed using neutral silica gel PSQ100B (Fuji Silysia Chemical) or silica gel 60 (Kanto Chemical).
  • Preparative recycling HPLC was performed using a YMC LC-forte / R equipped with a reverse phase column (YMC-Actus Triart C18). Unless otherwise stated, all reactions were performed under a nitrogen atmosphere.
  • compound 2 was obtained by the following method.
  • Bis [2-bromo-4- (N, N-diallylamino) phenyl] methane (Compound 1) (0.978 g, 1.89 mmol) in dehydrated THF (9 mL) at -78 ° C with s-butyl Lithium in cyclohexane and hexane (0.99 M, 4.00 mL, 3.96 mmol) was added dropwise over 5 minutes.
  • dichlorophenylphosphine PhPCl 2 ; 0.290 mL, 0.383 g, 2.14 mmol
  • compound 5 was also obtained by the following method. NaNO 2 (171 mg, 2.48 mmol) was added in air at 0 ° C. to a 96% sulfuric acid (2.5 mL) solution of compound 4 (247 mg, 0.739 mmol) in air. After stirring for 3 hours, the mixture was slowly added dropwise to ice and stirred at 110 ° C. for 0.5 hours. The resulting precipitate was collected by filtration, washed with distilled water, and dispersed in methanol. The obtained dark brown precipitate was filtered, and the filtrate was concentrated under reduced pressure to obtain 158 mg (0.470 mmol, yield 64%) of Compound 5 as a yellow solid.
  • the spectral data of Compound 6 is as follows.
  • compound 6 was also obtained by the following method.
  • Compound 5 (229 mg, 0.680 mmol) and imidazole (232 mg, 3.40 mmol) against dehydration CH 2 Cl 2 (25 mL) solution of, t- butyl chloro dimethyl silane (512 mg, 3.40 mmol) dehydrated CH 2 of A solution of Cl 2 (7 mL) was added. After stirring for 3 hours, water was added and the two layers were separated. The aqueous layer was extracted with CH 2 Cl 2 . The combined organic layers were dried over dehydrated sodium sulfate and filtered.
  • the obtained solid was dissolved in toluene, put into a separatory funnel, 1N HCl aqueous solution was added, and the separatory funnel was shaken.
  • the combined organic layers were washed with saturated brine and then dried over anhydrous sodium sulfate.
  • Sodium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to obtain Compound POF as a red solid in 30 mg (0.0730 mmol, 18% yield).
  • the spectral data of the compound POF is as follows.
  • the compound POF was also obtained by the following method. To a dehydrated THF (3 mL) solution of 2-bromotoluene (89 ⁇ L, 0.74 mmol), a cyclohexane and hexane solution (0.99 M, 0.90 mL, 0.89 mmol) of s-BuLi was added at ⁇ 78 ° C. The mixture was stirred at ⁇ 78 ° C. for 2 hours and a solution of compound 6 (139 mg, 0.246 mmol) in dehydrated THF (3 mL) was added slowly. The temperature was raised to room temperature and the reaction mixture was stirred for 1.5 hours. Thereafter, 0.5 M hydrochloric acid (20 mL) was added to quench the reaction.
  • the compound POF (37 mg, 0.090 mmol) obtained in Example 1 was dissolved in 5 mL of dry pyridine.
  • Acetic anhydride (85 ⁇ L, 0.90 mmol) was added at room temperature and the mixture was stirred for 3 hours.
  • the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (CH 2 Cl 2 / ethyl acetate 3/1 to 2/1), then further purified by recrystallization from CH 2 Cl 2 / hexane. Then, 33 mg (0.073 mmol, yield 81%) of compound AcPOF was obtained as a yellow powder.
  • the spectral data of the compound AcPOF is as follows. Mp. 175-179 ° C.
  • Synthetic compound 8 (800 mg, 1.57 mmol), 1,3-dimethylbarbituric acid (1.14 g, 7.30 mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh 3 ) 4 ; 1.01 g, 0.874 mmol was dissolved in degassed dehydrated 1,2-dichloroethane (16.0 mL), stirred at 85 ° C. for 3 days, and extracted with CH 2 Cl 2 . The combined organic layers were washed with saturated brine and then dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a crude product.
  • compositional organisms obtained under a nitrogen atmosphere and imidazole (85.0 mg, 1.25 mmol) were dissolved in dehydrated CH 2 Cl 2 (21 mL) and stirred for 30 minutes.
  • water was added and extracted with CH 2 Cl 2 .
  • the combined organic layers were washed with saturated brine and then dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a crude product.
  • the obtained solid was dissolved in toluene, placed in a separatory funnel, and extracted with 1N aqueous HCl.
  • the combined organic layers were washed with saturated brine and then dried over anhydrous sodium sulfate.
  • Sodium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to obtain Compound PSF as a red-purple solid in 18 mg (0.0422 mmol, yield 32%).
  • the spectral data of the compound PSF is as follows.
  • Example 1 LUMO and HOMO
  • the phosphafluorescein compound obtained in Example 1 (position 10 is a phosphorus atom), a known fluorescein compound (position 10 is an oxygen atom; TokyoGreen; JACS, 2005, 127, 4888), and a known silicon-substituted fluorescein compound (position 10)
  • the HOMO level and the LUMO level were calculated by structural optimization using the Gaussian 09 program. The calculation was performed at the B3LYP / 6-31 + G level. The results are shown in Table 1.
  • the phosphafluorescein compound of the present invention can reduce LUMO and HOMO (particularly LUMO) as well as the energy gap as compared with conventional compounds. For this reason, it is expected that the stability to light is further improved.
  • Example 2 X-ray crystal structure analysis
  • position 10 is a phosphorus atom
  • the structure was determined by the direct method (SIR-2003) and by the full matrix least squares method of F 2 (SHELXL-2013).
  • the crystal data is as follows.
  • the fluorescence spectrum was measured with a Hitachi F-4500 spectrometer with a resolution of 1 nm using a sample solution dissolved with 10 ⁇ 6 M.
  • the excitation spectrum was measured with a Horiba SPEX Fluorolog 3 spectrofluorometer equipped with Hamamatsu PMA R5509-73 and cooling system C9940-01, using a sample solution dissolved with 10 ⁇ 6 M.
  • the absolute fluorescence quantum yield was measured by Hamamatsu photonics PMA-11.
  • the phosphafluorescein compound obtained in Example 1 is dissolved in dimethyl sulfoxide (DMSO) as a solvent, and then the citric acid aqueous solution adjusted to pH 3 and Na 2 HPO are used.
  • DMSO dimethyl sulfoxide
  • a test solution in which the phosphafluorescein compound obtained in Example 1 was diluted to 7.4 ⁇ 10 ⁇ 6 M was prepared by diluting 100 times with a mixed aqueous solution of 4 aqueous solutions (pH 3 test solution).
  • test solution was prepared by diluting 100 times with a mixed aqueous solution of the aqueous solutions and dissolving the phosphafluorescein compound obtained in Example 1 so as to be 7.4 ⁇ 10 ⁇ 6 M (test solution of pH 7).
  • Example 1 when using a pH 9 buffered aqueous solution, the phosphafluorescein compound obtained in Example 1 was dissolved in dimethyl sulfoxide (DMSO), and then a mixed aqueous solution of Na 2 CO 3 aqueous solution and NaHCO 3 aqueous solution adjusted to pH 9 A test solution was prepared by diluting the phosphafluorescein compound obtained in Example 1 to 7.4 ⁇ 10 ⁇ 6 M (pH 9 test solution).
  • DMSO dimethyl sulfoxide
  • the absorption maximum wavelength and the fluorescence maximum wavelength can be made 600 to 700 nm, and the fluorescence quantum yield can be particularly improved.
  • the relative absorption at 627 nm increases with increasing pH and is almost the same in the neutral to alkaline region.
  • 627 nm with increasing pH. It can also be understood from the fact that the ratio of the excitation intensity at 532 nm to the excitation intensity at 532 nm is almost the same in the neutral to alkaline region. This behavior is considered to be due to the anionization of the phosphafluorescein compound of the present invention by increasing the pH.
  • Test Example 4 Photophysical characteristics (2)
  • the photophysical properties of test solutions 2 and 3 obtained in Test Example 3 above are reported values (TokyoGreen: JACS, 2005, 127, 4888, TokyoMagenda: Chem. Commun. 2011, 47, 4162, Nap-Fluorescein: Cytometry. 1989, 10, 151).
  • the results are shown in Table 2.
  • the phosphafluorescein compound of the present invention has the highest fluorescence quantum yield among compounds having an absorption maximum wavelength and a fluorescence maximum wavelength at 600 nm or more.
  • the absorption maximum wavelength is maximum, and self-absorption of organelles and light damage to cells can be minimized in bioimaging.
  • Test Example 5 Stability to light
  • DMSO dimethyl sulfoxide
  • a mixed aqueous solution of Na 2 HPO 4 solution and Na 2 HPO 4 solution adjusted to pH 7 was added and diluted 100 times.
  • a test solution 4 was obtained in which the obtained phosphafluorescein compound was dissolved to 7.4 ⁇ 10 ⁇ 6 M.
  • test solutions 4, 5 and 6 are irradiated with white light having a wavelength of 350 nm or more using a xenon lamp, and the absorption maximum wavelength (test solution 4: 627 nm; test solution 5: 491 nm; test solution 6: The absorbance maintenance rate at 582 nm was measured.
  • the results are shown in FIG.
  • the upper line (along with POF) is the phosphafluorescein compound of the present invention
  • the lower line (along with TM) is a known silicon-containing fluorescein compound
  • the line between them (along with TG) is known. It is a fluorescein compound.
  • Phosphorfluorescein dye has higher light stability than Cy5, which is a well-known representative red fluorescent dye, and is known as Alexa Fluor (registered trademark) 633, which is known as a red fluorescent dye excellent in light stability. It was shown to have the same light stability as Alexa Fluor (registered trademark) 647.
  • the compound AcPOF of Example 2 is 5 ⁇ M, 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (HEPES; pH 7.4) is 10 mM, DMSO. Was used, and DMEM containing Pluronic F-127 0.02% was used. The cells were stained at 37 ° C.
  • the phosphafluorocein compound of the present invention does not permeate the nuclear membrane, but permeates the cell membrane and enters the cell, and can be used as a fluorescent dye of the cell (HeLa cell etc.). .
  • Cytotoxicity assessment HeLa cells were seeded in flat bottom 96-well plates (1 ⁇ 10 4 cells / well) and cultured in DMEM containing 10% FBS at 37 ° C. in an incubator with 5% CO 2 /95% air for 24 hours. . The medium was then replaced with medium having various concentrations of compound AcPOF of Example 2 (1 ⁇ M, 5 ⁇ M and 10 ⁇ M) and the cells were further incubated at 37 ° C. for 24 hours.
  • 3- (4,5-di-methylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) reagent is added to each well (final concentration 0.5 mg / mL) and the plate was incubated for an additional 4 hours in a CO 2 incubator. Excess MTT in tetrazolium was removed and the cells were washed once with PBS. Formazan crystals were solubilized in DMSO (100 ⁇ L / well) at room temperature for 30 minutes, and then the absorbance of each well was measured with SpectraMax i3 (Molecular Devices) at a wavelength of 535 nm.
  • MTT 3- (4,5-di-methylthiazol-2-yl) -2,5-diphenyltetrazolium bromide
  • results are shown in FIG. In FIG. 7, the results represent the survival rate as a percentage relative to the case where no fluorescent dye was used. All data are shown by mean standard deviation (number of measurements n is 12). From this result, it can be understood that the phosphafluorocein compound of the present invention can cause cells to fluoresce within a range that can significantly reduce damage to the cells.

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Abstract

L'invention concerne un composé de phospha-fluorescéine représenté par la formule générale (I), ou un sel, un hydrate ou un solvate du composé de phospha-fluorescéine, qui est capable d'atteindre un rendement quantique suffisant, tout en présentant une longueur d'onde d'absorption maximale et une longueur d'onde de fluorescence maximale dans la plage de 600 à 700 nm. Ce composé de phospha-fluorescéine, ou son sel, son hydrate ou son solvate est également capable de réduire suffisamment HOMO. (Dans la formule (I), R1 représente un groupe aryle éventuellement substitué; R2 représente un atome d'hydrogène ou un groupe organique; et R représente un groupe qui est représenté par la formule (II) dans laquelle R3 représente un groupe aryle éventuellement substitué, un groupe alkyle éventuellement substitué, un groupe alcényle éventuellement substitué ou un groupe alcynyle éventuellement substitué; et R4 représente un groupe alkyle éventuellement substitué.
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018164252A1 (fr) * 2017-03-09 2018-09-13 富士フイルム株式会社 Composition, substance dichroïque, film anisotrope absorbant la lumière, stratifié et dispositif d'affichage d'images
WO2018181529A1 (fr) * 2017-03-28 2018-10-04 国立大学法人名古屋大学 Composé de phospha-rhodol, sel de celui-ci, et colorant fluorescent l'utilisant
CN110204575A (zh) * 2019-07-10 2019-09-06 郑州大学 含有芳香多环体系的磷杂六元环化合物及其合成方法
WO2020098138A1 (fr) * 2018-11-15 2020-05-22 武汉华星光电半导体显示技术有限公司 Matériau à fluorescence retardée thermo-activée bleu foncé et son application
CN112500714A (zh) * 2020-09-25 2021-03-16 四川大学 基于磷原子取代罗丹明衍生物骨架的染色试剂及其制备方法和应用
CN114716479A (zh) * 2022-05-05 2022-07-08 广州青苗新材料科技有限公司 一种具有热活化延迟荧光性质的膦氧类化合物及其制备与应用

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006257427A (ja) * 2000-02-07 2006-09-28 Applera Corp 電子欠乏窒素複素環で置換したフルオレセイン色素
JP2011521635A (ja) * 2008-05-27 2011-07-28 ダコ・デンマーク・エー／エス 新規なハイブリダイゼーションバッファーを用いた染色体異常の検出のための組成物及び方法
JP2013520962A (ja) * 2010-02-26 2013-06-10 ヴェンタナ メディカル システムズ， インク． ポリタグプローブ
CN103183710A (zh) * 2011-12-31 2013-07-03 昆山维信诺显示技术有限公司 一种膦氧基氢蒽二-三芳胺类化合物及其应用
US8506655B1 (en) * 2012-08-02 2013-08-13 Enzo Life Sciences, Inc. Fluorescent dyes containing phosphorus or arsenic
WO2014106957A1 (fr) * 2013-01-07 2014-07-10 国立大学法人 東京大学 SYNTHÈSE DE RHODAMINE Si ASYMÉTRIQUE ET DE RHODOL
JP2014529586A (ja) * 2011-08-10 2014-11-13 メルク パテント ゲーエムベーハー 金属錯体

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006257427A (ja) * 2000-02-07 2006-09-28 Applera Corp 電子欠乏窒素複素環で置換したフルオレセイン色素
JP2011521635A (ja) * 2008-05-27 2011-07-28 ダコ・デンマーク・エー／エス 新規なハイブリダイゼーションバッファーを用いた染色体異常の検出のための組成物及び方法
JP2013520962A (ja) * 2010-02-26 2013-06-10 ヴェンタナ メディカル システムズ， インク． ポリタグプローブ
JP2014529586A (ja) * 2011-08-10 2014-11-13 メルク パテント ゲーエムベーハー 金属錯体
CN103183710A (zh) * 2011-12-31 2013-07-03 昆山维信诺显示技术有限公司 一种膦氧基氢蒽二-三芳胺类化合物及其应用
US8506655B1 (en) * 2012-08-02 2013-08-13 Enzo Life Sciences, Inc. Fluorescent dyes containing phosphorus or arsenic
WO2014106957A1 (fr) * 2013-01-07 2014-07-10 国立大学法人 東京大学 SYNTHÈSE DE RHODAMINE Si ASYMÉTRIQUE ET DE RHODOL

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHAI, XIAOYUN ET AL.: "Near-Infrared Phosphorus- Substituted Rhodamine with Emission Wavelength above 700 nm for Bioimaging", CHEM. EUR. J., vol. 21, 30 September 2015 (2015-09-30), pages 16754 - 16758 *
SHINJI SUDA ET AL.: "Sekishoku Hakko o Shimesu Phospha-fluorescein Shikiso no Kaihatsu to Saibo Imaging eno Oyo", THE 95TH ANNUAL MEETING OF THE CHEMICAL SOCIETY OF JAPAN IN SPRING (2015) KOEN YOKOSHU IV, 11 March 2015 (2015-03-11), pages 1463 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018164252A1 (fr) * 2017-03-09 2018-09-13 富士フイルム株式会社 Composition, substance dichroïque, film anisotrope absorbant la lumière, stratifié et dispositif d'affichage d'images
CN110461951A (zh) * 2017-03-09 2019-11-15 富士胶片株式会社 组合物、二色性物质、吸光各向异性膜、层叠体及图像显示装置
JPWO2018164252A1 (ja) * 2017-03-09 2020-04-23 富士フイルム株式会社 組成物、二色性物質、光吸収異方性膜、積層体および画像表示装置
CN110461951B (zh) * 2017-03-09 2022-04-08 富士胶片株式会社 组合物、二色性物质、吸光各向异性膜、层叠体及图像显示装置
WO2018181529A1 (fr) * 2017-03-28 2018-10-04 国立大学法人名古屋大学 Composé de phospha-rhodol, sel de celui-ci, et colorant fluorescent l'utilisant
WO2020098138A1 (fr) * 2018-11-15 2020-05-22 武汉华星光电半导体显示技术有限公司 Matériau à fluorescence retardée thermo-activée bleu foncé et son application
CN110204575A (zh) * 2019-07-10 2019-09-06 郑州大学 含有芳香多环体系的磷杂六元环化合物及其合成方法
CN110204575B (zh) * 2019-07-10 2021-07-23 郑州大学 含有芳香多环体系的磷杂六元环化合物及其合成方法
CN112500714A (zh) * 2020-09-25 2021-03-16 四川大学 基于磷原子取代罗丹明衍生物骨架的染色试剂及其制备方法和应用
CN112500714B (zh) * 2020-09-25 2021-10-22 四川大学 基于磷原子取代罗丹明衍生物骨架的染色试剂及其制备方法和应用
CN114716479A (zh) * 2022-05-05 2022-07-08 广州青苗新材料科技有限公司 一种具有热活化延迟荧光性质的膦氧类化合物及其制备与应用
CN114716479B (zh) * 2022-05-05 2024-01-05 广州青苗新材料科技有限公司 一种具有热活化延迟荧光性质的膦氧类化合物及其制备与应用

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