CN112679543B - Lipophilic fluorescein probe and preparation method thereof - Google Patents
Lipophilic fluorescein probe and preparation method thereof Download PDFInfo
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Abstract
The application relates to the field of lipophilic fluorescein probes, and particularly discloses a lipophilic fluorescein probe and a preparation method thereof. The lipophilic fluorescein probe according to the present application is represented by the following chemical formula 1, wherein R is alkylene or cycloalkylene of C1-C20; r' is C1-C20 alkyl, cycloalkyl, aralkyl, aryl cycloalkyl, alkaryl or cycloalkaryl. The lipophilic fluorescein probe has high stability, is not easy to self-hydrolyze, can be rapidly hydrolyzed under the action of lipase to release fluorescein, and therefore has very high affinity action force and very low detection limit on the lipase. In addition, the preparation method is simple to operate and high in yield. [ chemical formula 1]
Description
Technical Field
The application relates to the field of lipophilic fluorescein probes, in particular to a lipophilic fluorescein probe and a preparation method thereof.
Background
Fluorescein, also known as: fluorescein, english name: fluoroescein, formula: c20H12O5Molecular weight: 332.31, CAS number: 2321-07-5. In 1871, bayer is first synthesized to obtain fluorescein by condensation of resorcinol and phthalic acid under the catalysis of zinc chloride, and because it has a high molar extinction coefficient, excitation and emission wavelengths in the visible light region, and has the advantages of high fluorescence quantum yield in water, no toxicity, low cost, etc., it is widely used in research of biochemistry, medicine, pharmaceutical chemistry, etc. Fluorescein belongs to xanthene dye, and has two isomers (shown as follows) of lactone type (yellow) and quinoid type (red), and the chemical names are respectively: 3 ', 6 ' -dihydroxyspiro [ isobenzofuran-1 (3H),9 ' - (9H) -xanthen-3-one]Or 9- (ortho-carboxyphenyl) -6-hydroxy-3H-xanthen-3-one. In its structure, an oxygen bridge bond fixes two benzene rings on one plane, so that the molecule has a coplanar rigid structure,can conjugate to generate delocalized big pi bond, so that strong fluorescence can be generated under the action of exciting light.
The fluorescein probe is an organic fluorescent functional dye which realizes the detection of target molecules by changing fluorescence spectra (including fluorescence intensity, fluorescence excitation and emission wavelength, fluorescence lifetime, fluorescence polarization/anisotropy and the like) after the fluorescein and the derivatives thereof react with the target molecules, has the advantages of extremely low detection limit, extremely high sensitivity and the like, and is particularly suitable for the related specific detection of certain low-content substances, particularly biological materials.
However, the molecular structure of fluorescein has two phenolic hydroxyl groups, which are strong in hydrophilicity, poor in lipophilicity and poor in affinity with cell membrane lipids, and the fluorescein is difficult to penetrate through cell membranes to enter the interior of cells, so that the application of the fluorescein in activity analysis of cells and enzymes is restricted. In order to improve the lipophilicity of fluorescein and make it easy to be absorbed by cells, those skilled in the art have found that the derivative obtained by esterifying two phenolic hydroxyl groups of fluorescein significantly improves its lipophilicity and is easy to permeate into cells, so that it can be used for studying the characteristics of cells and the cell environment. And, although the esterified fluorescein derivative is non-fluorescent, it is hydrolyzed by lipase to release fluorescein after entering the cells, so that the cells fluoresce, and the activity of the lipase is analyzed by detecting the intensity of the fluorescence. The gefengyan (synthesis and characterization of fluorescein derivatives [ D ] institute of pharmaceutical science and technology of Tianjin university, 2005) reports that a series of carboxylic fluorescein ester derivatives with 2-18 carbon atoms are synthesized and are applied to detection and analysis of lipase, and the relationship between the length of a carboxyl carbon chain in the fluorescein ester and the hydrolysis performance of the fluorescein ester is obtained so as to screen a fluorescein probe with better performance for detecting the lipase. However, there is room for further improvement in affinity for lipase and in minimum detection limit of the fluorescein probe.
Therefore, it would be desirable to develop lipophilic fluorescein probes with higher affinity for lipase and lower detection limits by those skilled in the art.
Disclosure of Invention
[ problem ] to
In view of the disadvantages of the prior art, an object of the present application is to provide a lipophilic fluorescein probe, which has no fluorescence, high stability, and is not easy to self-hydrolyze, and can rapidly hydrolyze to release fluorescein under the action of lipase, so that the lipophilic fluorescein probe has very high affinity to lipase and very low detection limit, and has a great application prospect.
An object of the present invention is to provide a method for preparing the lipophilic fluorescein probe, which is simple to operate and has high yield.
[ solution ]
In order to accomplish the above object, according to one embodiment of the present application, there is provided a lipophilic fluorescein probe, which is represented by the following chemical formula 1:
[ chemical formula 1]
Wherein R is alkylene or cycloalkylene of C1-C20;
r' is C1-C20 alkyl, cycloalkyl, aralkyl, aryl cycloalkyl, alkaryl or cycloalkaryl.
In the application, a phosphono hydrocarbon group is introduced to two hydroxyl groups of fluorescein, so that a fluorescein probe with very good lipophilicity is prepared, and particularly the phosphono hydrocarbon group is close to the phospholipid structure of a cell membrane, so that the penetration is excellent, and the fluorescein probe can very smoothly penetrate the cell membrane to enter cytoplasm. In addition, the fluorescein probe of the application also has very high affinity action force on lipase in cells, the Km is very small, the enzymatic reaction is very easy to carry out, so that fluorescein can be hydrolyzed to release and fluorescence can be detected, and the fluorescein probe has very high responsiveness to the lipase and extremely low detection limit.
Further, in the lipophilic fluorescein probe according to the present application, R may be preferably alkylene or cycloalkylene of C1 to C10.
More preferably, R may be a C2 to C8 alkylene or cycloalkylene group.
Still more preferably, R may be
Wherein A represents the end of the residue linked to a carbonyl group and B represents the end of the residue linked to a phosphono group.
By defining the above groups, the lipophilic fluorescein probe according to the present application has a higher affinity for lipase and a lower detection limit.
Further, in the lipophilic fluorescein probe according to the present application, R' may preferably be an alkyl group or an aralkyl group of C1 to C10.
More preferably, R' may be preferably an alkyl group or an aralkyl group of C2 to C8.
Still more preferably, R' may be
Wherein denotes a residue end to which a phosphinyloxy group is bonded.
By defining the above groups, the lipophilic fluorescein probe according to the present application has better lipophilicity and affinity for cell membrane phospholipids.
According to another embodiment of the present application, there is provided a method for preparing the lipophilic fluorescein probe, which includes the steps of:
(1) slowly adding an acyl chlorination reagent into Br-R-COOH, reacting at room temperature for 2-5 hours, and distilling under reduced pressure after the reaction is finished;
(2) adding fluorescein into dichloromethane, slowly adding the obtained substance in the step (1) under stirring at room temperature, then heating and refluxing for reaction for 2-6 hours, washing with a saturated sodium bicarbonate aqueous solution after the reaction is finished, separating liquid, distilling an organic phase under reduced pressure, washing the residue with alcohol, and drying;
(3) mixing the product obtained in the step (2) with P (OR')3And dispersing NaI and NaI in dioxane, carrying out reflux reaction for 4-9 hours, carrying out reduced pressure distillation, and purifying by column chromatography.
In this application, phosphono groups are introduced by first converting bromo carboxylic acid to bromo acid chloride, then reacting the bromo acid chloride with fluorescein to esterify the two phenolic hydroxyl groups of the fluorescein, and finally adding a trihydrocarbyl phosphite to replace the bromine element in the molecule. The preparation method has the advantages of stable reaction, simple operation and high yield.
The step (1) is an acyl chlorination reaction, and the reactivity of bromocarboxylic acid and the phenolic hydroxyl of fluorescein is improved by converting bromocarboxylic acid into bromoacyl chloride.
Preferably, in the step (1), the weight ratio of Br-R-COOH to the acyl chlorination reagent is 1 (3-4). By adding a sufficient excess of the acid chloride reagent, Br-R-COOH can be completely converted to bromoacid chloride Br-R-COCl.
Preferably, the acid chloride reagent in step (1) is thionyl chloride or oxalyl chloride. The product obtained after the reaction of thionyl chloride or oxalyl chloride is a gas, and is easily removed from the reaction system, and both of them themselves have a low boiling point, and therefore, they can be removed by distillation under reduced pressure after the completion of the reaction.
In addition, the reduced pressure distillation in the step (1) is carried out at 70-80 ℃. At this temperature, it is possible to facilitate the complete removal of the excess acid chlorinating reagent.
The step (2) is a reaction of acyl chloride and phenolic hydroxyl, and the fluorescein molecule can be easily esterified by adopting bromoacyl chloride with higher reaction activity.
Preferably, in the step (2), the molar ratio of the fluorescein to the product obtained in the step (1) is (1.05-1.25): 1. The bromine chloride in the step (1) can be completely reacted through the excessive fluorescein, and the residual fluorescein can be removed by water washing by utilizing the water solubility of the fluorescein.
In addition, the weight ratio of the fluorescein to the dichloromethane can be 1 (8-10).
And (3) carrying out reduced pressure distillation at 35-45 ℃ in the step (2). Beyond this temperature range, there is a risk of bumping.
The alcohol may be at least one selected from methanol, ethanol, or n-propanol.
The drying can be carried out at 55-65 ℃ for 3-4 hours.
Said step (3) belongs to the Michell-Albuzov (Michaelis-Arbuzov) reaction, P (OR')3Can react with the brominated hydrocarbon end of the obtained product in the step (2) to bond the two. Where NaI is added to further catalyze the promotion of the reaction.
Preferably, in the step (3), the product of the step (2), P (OR')3The molar ratio of NaI to NaI is 1 (1.1-1.3) to 0.03-0.05. Under the above molar ratio range, the reaction can be fully promoted, and the final product with higher yield can be obtained.
The weight ratio of the product obtained in the step (2) to dioxane may be 1 (8-10).
And (3) carrying out reduced pressure distillation at 60-70 ℃ in the step (2).
Preferably, in the step (3), the eluent for the column chromatography purification is ethyl acetate-petroleum ether-2: 1. Purifying by column chromatography to obtain lipophilic fluorescein probe product with high purity.
[ advantageous effects ]
In summary, the present application has the following beneficial effects:
the lipophilic fluorescein probe according to the present application has high stability, is not easily self-hydrolyzed, and can be rapidly hydrolyzed to release fluorescein under the action of lipase, thereby having very high affinity action force and very low detection limit to the lipase. In addition, the preparation method of the lipophilic fluorescein probe is simple to operate and high in yield.
Detailed Description
In order to make the present application more clearly understood by those skilled in the art, the present application is described in further detail with reference to the following examples, but it should be understood that the following examples are only preferred embodiments of the present application, and the scope of the present application as claimed is not limited thereto.
Sources of materials
Oxalyl chloride, thionyl chloride, fluorescein, lipase, available from Bailingwei technologies, Inc. of Beijing,
NaI, available from Shanghai Aladdin Biotechnology Ltd,
5-bromo-4-methylpentanoic acid, 6-bromohexanoic acid, 6-bromo-5-methylhexanoic acid, 2- (4- (bromomethyl) cyclohexyl) acetic acid, tris (3, 3-dimethylbutyl) phosphite, triphenylethyl phosphite, tris (2-methylbutyl) phosphite, and was designed from Tianjin Min concur medicine development Co., Ltd.
< example >
Example 1
The lipophilic fluorescein probe according to the present application was prepared using the following method:
(1) slowly adding 35g of oxalyl chloride into 10g of 5-bromo-4-methylpentanoic acid, reacting at room temperature for 5 hours, and distilling at 80 ℃ under reduced pressure after the reaction is finished to obtain 10.33g of product 5-bromo-4-methylpentanoyl chloride;
(2) adding 17.68g (0.05321mol) of fluorescein into 110mL (145.8g) of dichloromethane, slowly adding the 5-bromo-4-methyl valeryl chloride obtained in the step (1) under stirring at room temperature, then heating and refluxing for 5 hours, washing with saturated aqueous sodium bicarbonate solution after the reaction is finished, separating liquid, distilling an organic phase under reduced pressure at 40 ℃, washing a residue with ethanol, and then drying at 55 ℃ for 3 hours to obtain 28.08g of a product, namely bis (5-bromo-4-methyl pentanoic acid) fluorescein ester;
(3) the product obtained in step (2), 15.73g (0.04908mol) of tris (3, 3-dimethylbutyl) phosphite and 0.1839g (1.23mmol) of NaI were dispersed in 240mL (247.9g) of dioxane, refluxed for 9 hours, distilled under reduced pressure at 60 ℃ and purified by column chromatography (ethyl acetate: petroleum ether ═ 2:1) to obtain 30.92g of a final product, a lipophilic fluorescein probe (purity 99.83%, total yield 58.73%).
Example 2
The lipophilic fluorescein probe according to the present application was prepared using the following method:
(1) slowly adding 30g of oxalyl chloride into 10g of 6-bromohexanoic acid, reacting at room temperature for 3.5 hours, and distilling at 75 ℃ under reduced pressure after the reaction is finished to obtain 10.11g of product 6-bromohexanoic acid chloride;
(2) adding 19.67g (0.05919mol) of fluorescein into 140mL (185.5g) of dichloromethane, slowly adding the 6-bromohexanoyl chloride obtained in the step (1) while stirring at room temperature, then heating and refluxing for 6 hours, washing with a saturated aqueous solution of sodium bicarbonate after the reaction is finished, separating, distilling the organic phase at 45 ℃ under reduced pressure, washing the residue with methanol, and then drying at 55 ℃ for 3.5 hours to obtain 26.27g of the product, namely bis (6-bromohexanoate) fluorescein ester;
(3) the product obtained in step (2), 19.63g (0.04976mol) of triphenylethyl phosphite and 0.2295g (1.53mmol) of NaI were dispersed in 210mL (216.9g) of dioxane, refluxed for 6 hours, distilled under reduced pressure at 70 ℃, and purified by column chromatography (ethyl acetate: petroleum ether ═ 2:1) to obtain 29.56g of the lipophilic fluorescein probe as a final product (purity 99.67%, total yield 51.94%).
Example 3
The lipophilic fluorescein probe according to the present application was prepared using the following method:
(1) slowly adding 40g of thionyl chloride into 10g of 6-bromo-5-methylhexanoic acid, reacting at room temperature for 2 hours, and distilling at 70 ℃ under reduced pressure after the reaction is finished to obtain 10.41g of a product, namely 6-bromo-5-methylhexanoic chloride;
(2) adding 15.96g (0.04803mol) of fluorescein into 100mL (132.5g) of dichloromethane, slowly adding the 6-bromo-5-methylhexanoyl chloride obtained in the step (1) under stirring at room temperature, then heating and refluxing for 2 hours, washing with a saturated aqueous solution of sodium bicarbonate after the reaction is finished, separating liquid, distilling an organic phase at 45 ℃ under reduced pressure, washing a residue with methanol, and then drying at 60 ℃ for 4 hours to obtain 26.71g of a product, namely bis (6-bromo-5-methylhexanoic acid) fluorescein ester;
(3) the product obtained in step (2), 13.12g (0.04485mol) of tris (2-methylbutyl) phosphite and 0.2801g (1.87mmol) of NaI were dispersed in 250mL (258.3g) of dioxane, refluxed for 6 hours, distilled under reduced pressure at 65 ℃, and purified by column chromatography (ethyl acetate: petroleum ether: 2:1) to obtain 28.88g of a final product, a lipophilic fluorescein probe (purity 99.91%, total yield 60.50%).
Example 4
The lipophilic fluorescein probe according to the present application was prepared using the following method:
(1) to 10g of 2- (4- (bromomethyl) cyclohexyl) acetic acid was slowly added 40g of oxalyl chloride, followed by reaction at room temperature for 3 hours, and after completion of the reaction, distillation under reduced pressure at 75 ℃ was carried out to obtain 9.62g of the product, 2- (4- (bromomethyl) cyclohexyl) acetyl chloride;
(2) 14.49g (0.04361mol) of fluorescein was added into 100mL (132.5g) of dichloromethane, the 2- (4- (bromomethyl) cyclohexyl) acetyl chloride obtained in the step (1) was slowly added under stirring at room temperature, and then heated under reflux for 3 hours, after the reaction was completed, the mixture was washed with a saturated aqueous solution of sodium bicarbonate, subjected to liquid separation, and the organic phase was distilled under reduced pressure at 35 ℃ and the residue was washed with n-propanol, and then dried at 65 ℃ for 3 hours to obtain 22.88g of bis (2- (4- (bromomethyl) cyclohexyl) acetic acid) fluorescein ester as a product;
(3) the product obtained in step (2), 12.95g (0.03283mol) of triphenylethyl phosphite and 0.2237g (1.49mmol) of NaI were dispersed in 220mL (227.2g) of dioxane, refluxed for 4 hours, and purified by column chromatography (ethyl acetate: petroleum ether ═ 2:1) under reduced pressure at 60 ℃ to obtain 28.48g of a lipophilic fluorescein probe as a final product (purity 99.71%, overall yield 56.34%).
Example 5
The lipophilic fluorescein probe according to the present application was prepared using the following method:
(1) slowly adding 40g of thionyl chloride into 10g of 2- (4- (bromomethyl) cyclohexyl) acetic acid, then reacting at room temperature for 4.5 hours, and distilling at 70 ℃ under reduced pressure after the reaction is completed to obtain 9.91g of product 2- (4- (bromomethyl) cyclohexyl) acetyl chloride;
(2) adding 15.58g (0.04689mol) of fluorescein into 110mL (145.7g) of dichloromethane, slowly adding the 2- (4- (bromomethyl) cyclohexyl) acetyl chloride obtained in the step (1) into the dichloromethane under stirring at room temperature, heating and refluxing for 6 hours, washing the obtained mixture with saturated sodium bicarbonate solution after the reaction is finished, separating the obtained solution, distilling the organic phase at 45 ℃ under reduced pressure, washing the residue with ethanol, and drying the washed residue at 55 ℃ for 3.5 hours to obtain 24.66g of the product bis (2- (4- (bromomethyl) cyclohexyl) acetic acid) fluorescein ester;
(3) the product obtained in step (2), 10.35g (0.03539mol) of tris (2-methylbutyl) phosphite and 0.1447g (0.97mmol) of NaI were dispersed in 200mL (206.6g) of dioxane, refluxed for 7 hours, and purified by column chromatography (ethyl acetate: petroleum ether ═ 2:1) at 70 ℃ under reduced pressure to obtain 25.24g of a final product, a lipophilic fluorescein probe (purity 99.65%, overall yield 56.36%).
Test examples
The hydrolysis performance of the lipophilic fluorescein probes prepared according to examples 1 to 5 of the present application under the action of lipase was determined with reference to gefeng swallow (synthesis and characterization of fluorescein derivatives [ D ]. department of pharmaceutical science and technology, university of tianjin, 2005) chapter 3.14.
Specifically, the lipophilic fluorescein probes were dissolved in methylcellulose to a concentration of 10-3moL/L of a lipophilic fluorescein probe to be detected solution, and determining the following items by using the to-be-detected solution: (1) and (3) natural hydrolysis: diluting the solution to be detected by 100 times with Tris buffer (pH 7.2), detecting the change rate of fluorescence intensity with time at excitation wavelength of 492nm with a fluorescence spectrophotometer, and expressing the result as the average change value of fluorescence unit per minute in 14 days; (2) lipase detection limit and mie constant Km: the fluorescence spectrophotometer was zeroed by diluting the appropriate amount of the above-mentioned solution 100-fold with tris buffer (pH 7.2), and then lipase solutions of different concentrations were added, and the rate of change in fluorescence intensity with time of the solution was measured at an emission wavelength of 515nm, with the concentration required for doubling the enzymatic rate from the blank rate as a detection limit, and the mie constant Km was measured according to the Lineweaver-Burk plot method. The results are shown in table 1 below.
[ Table 1]
As can be seen from table 1 above, the lipophilic fluorescein probes prepared according to examples 1 to 5 of the present application all had very low or even negligible natural hydrolysis rates, indicating that they had very high stability and could be stably stored for a long time; in addition, the lipophilic fluorescein probe also has a very low detection limit for lipase, indicating excellent sensitivity, and a low mie constant indicating very high affinity for lipase, thereby greatly facilitating the enzymatic reaction.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.
Claims (10)
1. A lipophilic fluorescein probe, which is represented by the following chemical formula 1:
[ chemical formula 1]
Wherein R is alkylene or cycloalkylene of C1-C20;
r' is C1-C20 alkyl, cycloalkyl, aralkyl, aryl cycloalkyl, alkaryl or cycloalkaryl.
2. The lipophilic fluorescein probe as claimed in claim 1, wherein R is C1-C10 alkylene or cycloalkylene.
3. The lipophilic fluorescein probe of claim 2, wherein R is
Wherein A represents the end of the residue linked to a carbonyl group and B represents the end of the residue linked to a phosphono group.
4. The lipophilic fluorescein probe as claimed in claim 1, wherein R' is C1-C10 alkyl or aralkyl.
5. The lipophilic fluorescein probe of claim 4, wherein R' is
Wherein denotes a residue end to which a phosphinyloxy group is bonded.
6. A method for preparing the lipophilic fluorescein probe as described in any one of claims 1 to 5, comprising the steps of:
(1) slowly adding an acyl chlorination reagent into Br-R-COOH, reacting at room temperature for 2-5 hours, and distilling under reduced pressure after the reaction is finished;
(2) adding fluorescein into dichloromethane, stirring at room temperature, slowly adding the obtained substance in the step (1), heating, refluxing and reacting for 2-6 hours, washing with a saturated sodium bicarbonate aqueous solution after the reaction is finished, separating liquid, distilling an organic phase under reduced pressure, washing the residue with alcohol, and drying;
(3) mixing the product obtained in step (2) with P (OR')3And dispersing NaI and NaI in dioxane, carrying out reflux reaction for 4-9 hours, carrying out reduced pressure distillation, and purifying by column chromatography.
7. The preparation method according to claim 6, wherein in the step (1), the weight ratio of Br-R-COOH to acyl chloride reagent is 1 (3-4),
the acyl chlorination reagent in the step (1) is thionyl chloride or oxalyl chloride.
8. The method according to claim 6, wherein in the step (2), the molar ratio of fluorescein to the product obtained in the step (1) is (1.05-1.25): 1.
9. The process according to claim 6, wherein in the step (3), the product obtained in the step (2), P (OR')3The molar ratio of NaI to NaI is 1 (1.1-1.3) to 0.03-0.05.
10. The method according to claim 6, wherein in the step (3), the eluent for the column chromatography purification is ethyl acetate-petroleum ether-2: 1.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101983194A (en) * | 2008-04-03 | 2011-03-02 | 港大科桥有限公司 | Fluorophore compounds |
| CN102947266A (en) * | 2010-04-09 | 2013-02-27 | 生物状态有限公司 | Method of analysing a cell or other biological material containing a nucleic acid |
| CN104034705A (en) * | 2013-03-06 | 2014-09-10 | 常州欣宏科生物化学有限公司 | Method for detecting activity of enzyme by fluorescence probe |
| CN107778317A (en) * | 2016-08-26 | 2018-03-09 | 华中师范大学 | Methoxyl group fluorescein derivative and preparation method thereof and carboxypeptidase y detection method and detection kit |
| CN111212917A (en) * | 2017-08-24 | 2020-05-29 | 联邦科学技术研究组织 | BRET sensor molecules for detecting hydrolases |
-
2020
- 2020-12-29 CN CN202011592735.XA patent/CN112679543B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101983194A (en) * | 2008-04-03 | 2011-03-02 | 港大科桥有限公司 | Fluorophore compounds |
| CN102947266A (en) * | 2010-04-09 | 2013-02-27 | 生物状态有限公司 | Method of analysing a cell or other biological material containing a nucleic acid |
| CN104034705A (en) * | 2013-03-06 | 2014-09-10 | 常州欣宏科生物化学有限公司 | Method for detecting activity of enzyme by fluorescence probe |
| CN107778317A (en) * | 2016-08-26 | 2018-03-09 | 华中师范大学 | Methoxyl group fluorescein derivative and preparation method thereof and carboxypeptidase y detection method and detection kit |
| CN111212917A (en) * | 2017-08-24 | 2020-05-29 | 联邦科学技术研究组织 | BRET sensor molecules for detecting hydrolases |
Non-Patent Citations (2)
| Title |
|---|
| 检测活性氧物种的氧杂蒽类光学探针的研究进展;陈巍等;《分析化学》;20120915(第09期);全文 * |
| 荧光素二磷酸酯的合成和荧光分光光度法分析碱性、酸性磷酸酯酶;龚国权等;《高等学校化学学报》;19920715(第07期);全文 * |
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