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WO2007105529A1 - Compose chimioluminescent et agent de marquage le contenant - Google Patents

Compose chimioluminescent et agent de marquage le contenant Download PDF

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Publication number
WO2007105529A1
WO2007105529A1 PCT/JP2007/054254 JP2007054254W WO2007105529A1 WO 2007105529 A1 WO2007105529 A1 WO 2007105529A1 JP 2007054254 W JP2007054254 W JP 2007054254W WO 2007105529 A1 WO2007105529 A1 WO 2007105529A1
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chemiluminescent
derivative
compound
hydrogen
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Japanese (ja)
Inventor
Koji Suzuki
Koji Yamada
Akemi Sato
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Kanagawa Academy of Science and Technology
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Kanagawa Academy of Science and Technology
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    • 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
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
    • C07F5/02—Boron compounds
    • C07F5/022—Boron compounds without C-boron linkages
    • 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
    • C09K11/07—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials having chemically interreactive components, e.g. reactive chemiluminescent compositions
    • 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
    • C09K2211/1007—Non-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
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10—Non-macromolecular compounds
    • C09K2211/1003—Carbocyclic compounds
    • C09K2211/1011—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
    • 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
    • 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
    • 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
    • C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms

Definitions

  • the present invention relates to a chemiluminescent compound and a labeling agent comprising the same.
  • Chemiluminescent compounds are widely used as labeling agents in various fields including immunoassay. Many of them are obtained by connecting a chemiluminescent substance having good performance such as a lucigenin derivative and a fluorescent dye such as rhodamine through a spacer (Patent Documents 1 to 13).
  • chemiluminescent materials have problems such as unsatisfactory luminous efficiency, low stability, and difficulty in synthesis.
  • labeling agents it is difficult to change the fluorescence emission wavelength greatly only by changing the substituent.
  • a fluorescent dye having a polonedipyrromethene skeleton is known (Patent Document 14).
  • no chemiluminescent compound in which boron dipyrromethene fluorescent dye is bound to other chemiluminescent groups is known.
  • Patent Document 1 International Publication No. W098 / 54574
  • Patent Document 2 Japanese Patent Laid-Open No. 9-5238
  • Patent Document 3 Japanese Patent Laid-Open No. 10-81659
  • Patent Document 4 Japanese Patent Laid-Open No. 2004-51490
  • Patent Document 5 Japanese Patent Application Laid-Open No. 2004-26665
  • Patent Document 6 Japanese Patent Laid-Open No. 2003-183641
  • Patent Document 7 Japanese Patent Laid-Open No. 2003-137824
  • Patent Document 8 Japanese Patent Laid-Open No. 2002-302483
  • Patent Document 9 JP 2001-281257 A
  • Patent Document 10 JP 2001-115155 A
  • Patent Document 11 Japanese Patent Laid-Open No. 2001-81455
  • Patent Document 12 Japanese Patent Laid-Open No. 2000-245499
  • Patent Document 13 Japanese Patent Laid-Open No. 2000-178550
  • Patent Document 14 US Patent No. 5,723,218
  • Patent Document 15 Japanese Patent Laid-Open No. 2003-379505
  • Patent Document 16 US Pat. No. 5,869,689
  • Non-Patent Document 1 A. Burghart, H. Kim, M. B. Welch, L. H. Thoresen, J. Reibenspies, K
  • Non-Patent Document 2 M. A. T. Rogers, Journal of the Chemical Society, 1943, page 596
  • An object of the present invention is to provide a novel chemiluminescent compound capable of greatly changing the fluorescence emission wavelength only by changing a substituent having high luminous efficiency, and a labeling agent comprising the same. It is.
  • the present invention provides a chemiluminescent compound having a structure represented by the following general formula [I].
  • R 2 , R 4 and R 6 is a chemiluminescent group, and the others are hydrogen or any group which does not inhibit the light emission of the compound, R 5 and R 7 Is Independently of each other, hydrogen or any group that does not inhibit the light emission of the compound, R 8 and R 9 are independently of each other fluorine, an alkoxy group, or a chemiluminescent group (however, both are not chemiluminescent groups at the same time) ).
  • the present invention also provides a labeling agent comprising the chemiluminescent compound of the present invention. Furthermore, the present invention provides a combination of labeling agents comprising a plurality of types of the chemiluminescent compounds of the present invention having different emission wavelength adjusting groups from each other and thereby different emission wavelengths. Furthermore, the present invention provides a method for measuring a labeled substance, which comprises subjecting the substance labeled with the labeling agent of the present invention to a reaction, and after the reaction, causing the labeling agent to emit light and measuring the substance.
  • the present invention uses a plurality of types of the chemiluminescent compounds of the present invention having different emission wavelength adjusting groups from each other and thereby different emission wavelengths as labeling agents, and each labeling with the plurality of labeling agents.
  • the present invention provides a method for simultaneously measuring a plurality of labeled substances, comprising: subjecting the plurality of kinds of substances to a reaction, and after the reaction, causing each of the labeling agents to emit light and simultaneously measuring the plurality of types of the substances.
  • a novel chemiluminescent compound capable of greatly changing the fluorescence emission wavelength only by changing a substituent having high emission efficiency and a labeling agent comprising the same are provided.
  • the boron dipyrromethene skeleton employed in the chemiluminescent compound of the present invention has high luminous efficiency.
  • the chemiluminescent group is not bonded to the boron dipyrromethene skeleton via a spacer. Since it is directly coupled, the luminous efficiency can be further increased.
  • the fluorescence emission wave The length can be changed greatly. Since introduction of an arbitrary substituent on the boron dipyrromethene skeleton can be easily performed by a conventional method, a plurality of chemiluminescent compounds having different fluorescence emission wavelengths can be easily synthesized according to the present invention. be able to. Since these structures may have the same structure other than the modified substituent, in particular, the chemiluminescent group that chemiluminescents by the reaction, these multiple chemiluminescent substances emit light optimally under the same conditions.
  • the plurality of types of substances are respectively the same.
  • it becomes possible to measure This makes it possible to measure a plurality of different types of test substances at the same time, for example, in an immunoassay, and thus provides a new measurement technique according to the present invention.
  • FIG. 1 is a diagram showing a spectrum of chemiluminescence wavelength of a compound of the present invention produced in an example of the present invention in comparison with a known dye.
  • the chemiluminescent compound of the present invention has a structure represented by the above general formula [I].
  • at least one of R 2 and R 6 is a chemiluminescent group, and other than that, hydrogen or any group that does not inhibit the light emission of the compound
  • R 5 and R 7 are each independently hydrogen or any group that does not inhibit the light emission of the compound
  • R 8 and R 9 are each independently fluorine, an alkoxy group or a chemiluminescent group (provided that both are simultaneously Not a chemiluminescent group).
  • the chemiluminescent group is preferably R 2 and / or R 6 .
  • the chemiluminescent group is a group having a structure that emits light by a chemical reaction.
  • the boron dipyrromethene moiety emits fluorescence by the light emitted from the chemiluminescent group, and this fluorescence is measured. Therefore, any chemiluminescent group that emits light that can excite the boron dipyrromethene moiety and emit fluorescence can be used as long as it does not necessarily emit visible light. Can do.
  • a luminescent group if it is used in this field as a labeling agent, it is! Any of these can be used, and various ones are known or well known.
  • Preferred examples include phthalcarbazide derivatives, dioxetane derivatives, oral fin derivatives, atalidine derivatives, indole derivatives, oxalic acid derivatives, diphenol derivatives or luciferin derivatives, all of which are well known in the art.
  • Each of these chemical luminescent groups is represented by the following general formula.
  • R is independently of each other hydrogen, an alkyl group, an aryl group, halogen, alkoxy, aryl alkyl, cycloalkyl, acyl, formyl, sialalkyl, heteroaryl, It is a substituent such as mono- or di-alkylamino, and one R in each general formula is bonded to the boron dipyrromethene skeleton directly or via a spacer.
  • the carbon number of the alkyl group in the group containing an alkyl group or an alkyl group part is usually about 1 to 10.
  • the term “alkyl group” includes both a linear alkyl group and a branched alkyl group.
  • any of the four Rs is -NR a R b (wherein R a R b is independently hydrogen or an alkyl group) is a luminol derivative.
  • a phthalhydrazide derivative capable of easily synthesizing a raw material phthalic acid capable of coupling synthesis to a boron dipyrromethene skeleton having a high emission quantum yield is preferable.
  • chemiluminescent groups are usually used in each compound, but a plurality of types may be combined.
  • the chemiluminescent group can be bonded to the boron dipyrromethene skeleton through a spacer such as an alkylene group, but by bringing the chemiluminescent group as close as possible to the boron dipyrromethene skeleton, a fluorescence that can generate partial dipyrromethene partial force is also generated.
  • the chemiluminescent group is directly bonded to the boron dipyrromethene skeleton without using a spacer. By selecting the boron dipyrromethene skeleton as the fluorescent dye, the chemiluminescent group can be easily coupled directly.
  • the boron dipyrromethene skeleton emits fluorescence
  • the boron dipyrromethene skeleton emits fluorescence.
  • groups other than the chemiluminescent group are not particularly limited. It may be any group that does not inhibit the light emission of the compound. Examples of the optional group include a light emission wavelength adjusting group, a binding group, and a polar group described later. As mentioned above, any group that does not fall under any of these may be hydrogen or any group that does not inhibit light emission of the compound, but preferred examples include a hydrogen atom and an alkyl group having 1 to 4 carbon atoms. be able to.
  • the fluorescence emission wavelength can be changed without changing other parts by changing the substituent bonded to the boron dipyrromethene skeleton. Can be greatly changed. It is preferable to have a light emission wavelength adjusting group as at least one of R 1 to R 7 utilizing this feature.
  • the “emission wavelength adjusting group” means a group in which the emission wavelength of fluorescence changes when this group is bonded as compared to the case where this group is not bonded.
  • the emission wavelength adjusting group is a group that can be combined with a boron dipyrromethene skeleton, and the wavelength of the fluorescence emitted by the boron dipyrromethene moiety.
  • the emission wavelength adjusting group may be any group.
  • the fluorescence wavelength changes relatively large by changing a single group.
  • the fluorescence wavelength may change on the longer wavelength side compared to the case where no emission wavelength adjusting group is provided. Therefore, preferred examples of the emission wavelength adjusting group include an alkyl group, an alkoxyl group, an aryl group, a heteroaryl group, and derivatives thereof.
  • carbon number of the alkyl part in an alkyl group or an alkoxyl group 1-10 are preferable.
  • a aryl group a phenol group and a naphthalene group are preferable.
  • a heteroaryl group one or more carbon atoms in the phenol group and the naphthalene group are nitrogen, oxygen, and Z or Groups substituted with iow are preferred.
  • a hydrogen atom bonded to one or a plurality of carbon atoms constituting these groups is a halogen, a cyano group, an alkoxyl group, an amino group, a hydroxyl group, a mono- or dialkyl.
  • a group such as an amino group, a carbo group, a carboxyl group, a thiol group, or a disulfide group are preferred, and the carbon number of the alkyl moiety in these substituents is preferably 1 to 10 forces, and 1 to 6 is preferred. Further preferred.
  • the above alkyl group or alkoxyl group is constituted. A group in which one or a plurality of carbon atoms formed is replaced with at least one of oxygen, nitrogen and nitrogen is also preferable as the derivative of the alkyl group or alkoxyl group.
  • the hydrogen atom bonded to one or more carbon atoms constituting these groups is halogen, cyano group, alkoxyl group, amino group, hydroxyl group, mono- Alternatively, those substituted with a group such as a di-alkylamino group, a carbonyl group, a carboxyl group, a thiol group, or a disulfide group are preferred.
  • the carbon number of the alkyl moiety in these substituents is preferably 1 to 10 1 ⁇ 6 is more preferred.
  • the emission wavelength adjusting group a substituent such as benzocrown, N-arylazacrown, amide group, etc., whose electronic structure is changed by molecular recognition or chemical reaction and whose emission wavelength is changed is used. .
  • the fluorescence wavelength of the boron dipyrromethene fluorescent dye is greatly influenced by the type of substituents bonded to R 1 and R 7 in the general formula [I]
  • the above-described emission wavelength adjusting group is represented by R 1 And Z or R 7 is preferred.
  • a plurality of emission wavelength adjusting groups can be bonded, and different types of emission wavelength adjusting groups can be bonded to one boron dipyrromethene skeleton.
  • each labeling agent must be subjected to the chemiluminescence reaction under the same conditions. Therefore, it is preferable that the chemiluminescent group of each labeling agent is the same.
  • At least one of R 1 to R 7 in the general formula [I] has a binding group.
  • the binding group means a group that can be used for binding to another compound. Since chemiluminescent labeling agents are often bound to biologically related substances such as proteins, polypeptides, and sugars, the binding groups include amino groups, hydroxyl groups, and carboxyls that are convenient for binding to these biologically related substances. Preferred examples include groups, sulfonic acid groups, thiol groups, disulfide groups, isocyanate groups, thioisocyanate groups, succinimidyl ester groups, pentafluorophenyl ester groups, maleimide groups, and the like.
  • these groups need only be used for bonding with other compounds, at least any group including these groups (for example, an aminoalkyl group, etc.) is also a bonding group. It can be used as One binding group is sufficient, and one binding site is specified. It is usually preferred to have one, but it may contain two or more linking groups.
  • At least one of R 1 to R 7 may contain a group for adjusting solubility. Since the chemiluminescent labeling agent is often used in an aqueous system, when it is desired to improve the solubility in the aqueous system by increasing the hydrophilicity of the chemiluminescent compound, R 1 to R 7 At least one of them may contain a hydrophilic group. Examples of such a hydrophilic group include a sulfonic acid group or a salt thereof, a hydroxyl group, an amino group, a carboxylic group, a carboxyl group, or an alkyl group containing at least one of these groups or one or more ether bonds.
  • Examples thereof include (but preferably have 1 to 4 carbon atoms) or alkoxyl groups (preferably 1 to 4 carbon atoms), but are not limited thereto. Further, such a hydrophilic group may be used in combination with the above-described binding group, emission wavelength adjusting group, and the like.
  • R 8 and R 9 are each independently a fluorine, alkoxyl group or the above-described luminescent group, and are preferably independently a fluorine or an alkoxyl group, and an alkoxy group.
  • the number of carbon atoms is preferably 1-6.
  • chemiluminescent compound of the present invention include a compound having a structure represented by the following general formula [la].
  • R la and R 7a are independently of each other hydrogen, an alkyl group, an alkoxyl group, a halogen, a phenol group, or a substituted phenol group (the substituent is 1 to 5 alkyl groups, alkoxyl group or a mono - or di - alkylamino group) (wherein, R la and both R 7a is not simultaneously hydrogen) or hetero aromatic group, R 2a is hydrogen, halogen, terminus binding functional group Substituted carbonyl group or aryl group having the above, emission wavelength adjusting group, hydrophilic R 3a and R 5a are each independently hydrogen or an alkyl group, R 1 ⁇ 2 is hydrogen, an alkyl group, a phenyl group, or a substituted alkyl group or aryl having a binding functional group at the terminal.
  • the phthalcarbazide derivative portion is a chemiluminescent group
  • R la and R 7a are emission wavelength adjusting groups (except for hydrogen)
  • R 3a and R 5a are hydrogen or an arbitrary substituent
  • R 1 ⁇ 2 Corresponds to hydrogen or any substituent or linking group.
  • R lb and R 7b are each independently an alkyl group having 1 to 4 carbon atoms, a phenol group or an alkoxy phenol group (the alkyl moiety has 1 to 10 carbon atoms),
  • R 2b is hydrogen or a sulfonic acid group or a salt thereof,
  • R 3b and R 5b are independently of each other hydrogen or an alkyl group having 1 to 4 carbon atoms,
  • R 4b is hydrogen or
  • R 1Q and R 11 are each independently hydrogen or an alkyl group having 1 to 4 carbon atoms, Represents an integer of 1 to 4).
  • the chemiluminescent compound of the present invention can be produced by bonding a chemiluminescent group to a boron dipyrromethene skeleton to which the above-described substituent is bonded, if necessary. Since the boron dipyrromethene skeleton and the chemiluminescent group are known and the production method thereof is also known, the chemiluminescent compound of the present invention can be easily produced by ordinary chemical synthesis. Some specific examples are described in detail in the examples below.
  • R 2 corresponds to R 1Q and R 11 in the substituent of R 4b in the general formula [lb], and R represents R 3b and R 11b in the general formula [lb].
  • Ar corresponds to R lb and R 7b in the general formula [lb].
  • dipyrromethene derivative VII When cyano-1,4-benzoquinone (DDQ) is sequentially added, the dipyrromethene derivative VII is produced.
  • Boron dipyrromethene derivative VIII is produced by adding trifluoroborane jetyl ether complex to toluene in the presence of triethylamine and refluxing. When this VIII is reacted with N-bromosuccinimide in black mouth form in the same manner as in Patent Document 16, bromine adduct IX is produced. In this reaction, bromine diadducts can be easily controlled by controlling the equivalent amount of NBS. Can be synthesized.
  • H, R2 CH3
  • chlorosulfonic acid in a methylene chloride solution.
  • the 8-position unsubstituted boron dipyrromethene XVII synthesized by the method described in Non-Patent Document 2 is treated in the same manner as in the synthesis route of II above, so that the 8-position unsubstituted chemistry of the present invention can be obtained in 4 steps.
  • the luminescent dye XV could also be synthesized.
  • the chemiluminescent compound of the present invention can be used as a chemiluminescent labeling agent.
  • the chemiluminescent labeling agent itself is well known, and the chemiluminescent compound of the present invention can also be used in the same manner as conventional chemiluminescent labeling agents.
  • the labeling substance is labeled by binding the chemiluminescent compound of the present invention to the labeling substance to be labeled.
  • the chemiluminescent compound of the present invention has the above-described binding group
  • the chemiluminescent compound of the present invention is bound to the labeling substance via the binding group by a conventional method.
  • a conventional method Do not have a linking group!
  • an intermediate having a binding group introduced can be prepared by a conventional method and bound to a labeling substance.
  • the mode of use of the chemiluminescent labeling agent is exactly the same as in the prior art.
  • the labeling substance (referred to herein as "labeling substance") ) Is subjected to a reaction, and a washing step or the like is performed as necessary. Then, the label is chemiluminescent and the luminescence is measured.
  • the labeling agent can be used as follows.
  • a primary antibody against the target antigen is bound to a solid phase and reacted with the test sample, so that the target antigen in the test sample is immobilized on the solid phase. Conjugated to the primary antibody. After washing, a second antibody labeled with a chemiluminescent labeling agent is reacted. After washing, the chemiluminescent labeling agent of the second antibody bound to the target antigen is chemiluminescent by a chemical reaction according to a conventional method, and luminescence is measured.
  • “measurement” includes any of detection, quantification, and semi-quantification.
  • the chemiluminescence of the labeling agent can be performed by conducting a chemical reaction according to the chemiluminescent group in each chemiluminescent labeling agent by a conventional method.
  • a chemiluminescent group is preferred, and an example phthalcarbazide derivative is used as a chemiluminescent group, as well known, in the presence of the catalyst hexocyano potassium (III) potassium (ferricyani potassium), hydrogen peroxide and By making it react, a chemiluminescent group can be light-emitted.
  • the boron dipyrromethene skeleton is excited by light from the chemiluminescent group, and fluorescence is emitted from the boron dipyrromethene skeleton. The fluorescence emission is measured.
  • the fluorescence wavelength can be greatly changed to be discernable by changing the above-described emission wavelength adjusting group. Bonding a substituent on the boron dipyrromethene skeleton is easily accomplished by conventional chemical synthesis. be able to. Therefore, a plurality of types of chemiluminescent compounds of the present invention having different fluorescence emission wavelengths can be easily produced. By using a plurality of types of chemiluminescent compounds of the present invention having different wavelengths of fluorescence emission so that they can be distinguished as chemiluminescent labeling agents, it becomes possible to label a plurality of types of substances with different chemiluminescent labeling agents.
  • a plurality of different test substances can be measured simultaneously.
  • different types of secondary antibodies can be labeled with different labeling agents so that the fluorescence wavelength can be distinguished.
  • the multiple types of labeling agents used need only differ so that the respective fluorescence emission wavelengths can be distinguished (that is, the emission wavelength ranges do not overlap or hardly overlap), and one of them is the emission wavelength adjustment. It may have no group. That is, the present invention has the emission wavelength adjusting groups different from each other.
  • the chemiluminescent labeling agent of the present invention can use a plurality of types of chemiluminescent labeling agents in which the chemiluminescent group and the skeleton of the fluorescent dye are identical to each other, and only the substituents on the boron dipyrromethene skeleton are different.
  • the chemiluminescent labeling agent of the present invention can be preferably used as a labeling agent in the dideoxy method when used for determining the base sequence of DNA.
  • V and 4 kinds of dideoxynucleotides are labeled with fluorescent dyes of different fluorescence wavelengths.
  • fluorescent dyes of different fluorescence wavelengths instead of these four types of fluorescent dyes, four types of labeling agents of the present invention having different emission wavelengths can be used.
  • the labeling agent of the present invention is used instead of the fluorescent dye. (1) Since a light source such as a laser is not required for detection, the structure of the measuring device is simplified and can be supplied at low cost.
  • the sensitivity is improved at least several times, so the amount of primer and dye-labeled dideoxynucleotide can be reduced, and the amplification time is shortened, so that measurement can be performed quickly.
  • Methylene chloride 100 mL was placed in a 300 mL three-necked flask and N-published for 30 minutes at room temperature.
  • ops was added and stirred at room temperature for 1 hour. The color changed from yellow to magenta.
  • DDQ (383 mg, 1.69 mmol, 1 eq) was added and stirred at room temperature for 30 minutes.
  • the reaction mixture was allowed to cool, and black mouth form and water were added to extract the organic phase.
  • the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was concentrated under reduced pressure.
  • H 0 130 mL
  • KOH 33.6 g
  • M 110Z / ⁇ I , ⁇ 600] ⁇ ⁇ ⁇ ⁇ ⁇ 0 os ⁇ nc ⁇ SZ) ⁇ 3 ⁇ 4 ⁇ ⁇ 9 S) N3 ⁇ 4 ⁇ ° ⁇ Y ⁇
  • the mixture was degassed again and stirred at 80 ° C for 1 day under Ar.
  • the mixture was degassed again and stirred at 80 ° C. for 24 hours under Ar.
  • the mixture was degassed again and stirred at 80 ° C for 24 hours under Ar.
  • Luminescence experiments were performed on newly synthesized KCB-lime2, orange2 and apple2.
  • ABEI aminobutylethylisoluminol

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
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Abstract

L'invention concerne un nouveau composé chimioluminescent présentant une efficacité lumineuse élevée, dont la longueur d'onde d'émission de fluorescence peut être modifiée dans une large mesure par simple remplacement d'un substituant par un autre. L'invention concerne également un agent de marquage contenant ce composé chimioluminescent. Le composé selon l'invention présente une structure de formule générale [I], dans laquelle au moins R2, R4 ou R6 représente un groupe chimioluminescent et les autres représentent indépendamment un groupe hydrogène ou tout autre groupe qui n'interfère pas dans l'émission lumineuse du composé; R1, R3, R5 et R7 représentent indépendamment un groupe hydrogène ou tout autre groupe qui n'interfère pas dans l'émission lumineuse du composé; et R8 et R9 représentent indépendamment un groupe fluor, un groupe alcoxy ou un groupe chimioluminescent (à condition qu'il soit impossible que R8 et R9 représentent ensemble un groupe chimioluminescent).
PCT/JP2007/054254 2006-03-07 2007-03-06 Compose chimioluminescent et agent de marquage le contenant Ceased WO2007105529A1 (fr)

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JP2006061786A JP2007238489A (ja) 2006-03-07 2006-03-07 化学発光性化合物及びそれから成る標識剤

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Cited By (5)

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WO2009056782A1 (fr) * 2007-10-29 2009-05-07 Ucl Business Plc Luminescence
WO2010098119A1 (fr) * 2009-02-27 2010-09-02 三井化学株式会社 Composé complexe et dispositif électroluminescent organique contenant le composé complexe
WO2010098098A1 (fr) * 2009-02-27 2010-09-02 出光興産株式会社 Composés du complexe pyrrométhène-bore et éléments organiques électroluminescents les utilisant
CN102333783A (zh) * 2009-02-27 2012-01-25 出光兴产株式会社 亚甲基吡咯硼配位化合物及使用了其的有机电致发光元件
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CN111139066A (zh) * 2019-12-24 2020-05-12 山东大学 一种广谱型低电位电化学发光体系及其构建方法
CN111139066B (zh) * 2019-12-24 2021-05-07 山东大学 一种广谱型低电位电化学发光体系及其构建方法
CN115298286A (zh) * 2020-03-20 2022-11-04 日东电工株式会社 含硼环状发光化合物和包含该含硼环状发光化合物的色彩转换膜

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