CN106544399A - The application of the high specific fluorescence probe of one class detection Cytochrome P450 1A1 - Google Patents
The application of the high specific fluorescence probe of one class detection Cytochrome P450 1A1 Download PDFInfo
- Publication number
- CN106544399A CN106544399A CN201611115346.1A CN201611115346A CN106544399A CN 106544399 A CN106544399 A CN 106544399A CN 201611115346 A CN201611115346 A CN 201611115346A CN 106544399 A CN106544399 A CN 106544399A
- Authority
- CN
- China
- Prior art keywords
- cyp1a1
- probe
- fluorescence
- high specific
- application
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The application of the high specific fluorescence probe of one class detection Cytochrome P450 1A1, which belongs to field of fine chemical.The Specific probe is resorufin class compound ether derivative, and which can be used for the quantitative determination of the presence or absence and its vigor for detecting CYP1A1 in different biological samples.It is as follows that enzyme activity specifically determines flow process：Select resorufin ether derivative hydrolysis to be probe reaction, select suitable substrate concentration in linear reaction interval by the quantitative determination unit interval in its hydrolysis metabolite resorufin growing amount come determine each biological sample, cell, in body and overall organ CYP1A1 enzymes substantial activity.The probe cannot be only used for the qualitative assessment of CYP1A1 enzyme activity in separate sources biological specimen, can be additionally used in the inhibitor of external quick screening CYP1A1 in addition by the probe reaction.
Description
Technical field
The present invention relates to the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1, which belongs to fine
Chemical field.
Background technology
Cytochrome P 450 enzymes (CYP450) are a MO superfamilies containing heme, are predominantly located at endoplasm
Net, is most important metabolic enzyme in body.These enzymes participate in including chemicals in carcinogenic substance, environment, medicine and many interior
The bioconversion of endogenous compound, to maintaining human normal physiological function to play an important role.The I of cytochrome P 450 enzymes catalysis
Phase reaction is the committed step of compound metabolism in vivo, because this single step reaction is typically medicine from the internal rate-limiting step removed
Suddenly, the dynamic characteristics such as half-life, the clearance rate of compound can be affected, and P450 enzymatic activitys is often with inherent cause, age, disease
Impact that state or other medicines interact and change.
Gene of the human genome comprising 57 CYP450, CYP superfamilies are divided into 18 families.Cytochrome P450 1A1
(CYP1A1) it is important I phase metabolic enzymes, mainly expresses in people's extrahepatic tissue, such as people's lung, skin, small intestine.CYP1A1 is front
Play in terms of the bioconversion of carcinogenic substance and poisonous substance (such as polycyclic aromatic hydrocarbon, heterocycle arylamine or acid amides, azo compound etc.) and focus on
Act on.Early-stage Study confirms, the oxidation of precarcinogen can generate fragrant amine oxide, glycol epoxide and its
His electrophilic reactive species, these materials further form adduct with DNA or protein, cause tumour to be formed.
In some specific crowds (such as smoker), the probability that said process occurs then is significantly increased.Additionally, CYP1A1 is also in some medicines
Key player is play in the metabolite clearance of (such as phenaetin, caffeine, Fa Hualin and other curative drugs).
It should be noted that the distribution of CYP1A1 has very big individual difference, the expression of CYP1A1 enzymes is by heredity, year
The impact of age, disease, sex, environment and many factors such as thing of taking medicine altogether.For example, for a long time with polycyclic aromatic hydrocarbon and heterocycle arylamine or
Acid amides, and come from the crowd of the aflatoxins contact of tobacco or deteriorating food, the expression of CYP1A1 and function can show
Write and lifted, cause contents of the CYP1A1 in Different Individual also to have very big difference, its content range is 0-3pmol/mg.
The invention provides the ether derivative of a class resorufin class compound and its as CYP1A1 enzyme fluorescence probes bottom
The application of thing, which can generate hydrolysate of the fluorescence emission spectrum different from prototype Jing after CYP1A1 hydrolysis.The enzymatic reaction has
The features such as selective high, metabolite is easily detected, enzyme activity and inhibitory activity are evaluated rapidly and efficiently.
The content of the invention
It is an object of the invention to provide the high specific fluorescence probe of class detection Cytochrome P450 1A1 (CYP1A1)
And its Fluorescence Fluorescence attribute of application, the substrate prototype and hydrolysate has notable difference.Can be to many using the probe reaction
The distribution and function for planting CYP1A1 in living things system carries out quantitative assessment.
It is an object of the invention to provide the high specific fluorescence probe of class detection Cytochrome P450 1A1 (CYP1A1)
And its application, the ehter bond of the substrate can be corresponding hydrolysate by CYP1A1 specific for hydrolysis in human tissue or cell, and
Product has fluorescence；The substrate is, with resorufin class compound as raw material, to obtain corresponding 7- hydroxyls ethers by esterification and spread out
Biology, shown in its general structure such as formula (1), wherein, R be methyl, ethyl, chloroethyl, n-propyl, normal-butyl, n-pentyl, just oneself
Base, phenyl, p-methylphenyl, to ethylphenyl, to propyl group phenyl, to hexyl propyl group, p-nitrophenyl, rubigan, 1- how
One kind in base, 2- furyls, 2- thienyls, (4- phenyl) phenyl, 4- ethoxyl phenenyl substituents.
Such as when R is chloroethyl base, the substrate is 7- chloroethoxy resorufins (CHPO).
Present invention also offers the application of the specificity fluorescent probe substrate of CYP1A1, the substrate is used as the special of CYP1A1
Property substrate, there is hydrolysis, enzyme or cell determined by the growing amount of hydrolysate in the quantitative determination unit interval and is prepared
The activity of CYP1A1 in the biological samples such as liquid and cell；Specifically assay method is：
--- the ether derivative in system using resorufin class compound 7- hydroxyls is used as Specific probe；Substrate is dense
Degree selects 1/10～10Km；Concentration of substrate preferred K during single point assaym。
--- in the conventional buffer solution such as PBS or Tris-HCl, reaction temperature is that preferably 37 DEG C are between 20 DEG C to 60 DEG C
The peak optimization reaction time；Between 5.5～10.5, preferred pH 7.4 is peak optimization reaction pH value to incubation system pH；
--- the reaction time is 5～120 minutes, is guaranteeing that the corresponding hydrolysate of above substrate reaches quantitative limit and substrate
Terminating reaction when conversion ratio is less than 20%；
--- evaluation index of the hydrolysate growing amount as CYP1A1 activity in the analytical unit time.
The application of the human-cytochrome P4501A1 specificity fluorescent probe substrates that the present invention is provided, the substrate elimination factor
Or the production rate of hydrolysate should be between 0.1%～20%.
The application of the human-cytochrome P4501A1 specificity fluorescent probe substrates that the present invention is provided, probe substrate and its water
Solution product is respectively provided with fluorescence properties, and the rapid sensitive detection of product and substrate can be realized using fluorescence detector；Fluoroscopic examination bar
Part is：480～530nm of excitation wavelength, carries out the detection of fluorescence emission spectrum in 560～630nm.Additionally, the specific probe bottom
Thing and corresponding CYP1A1 Activity determinations process will not be disturbed by living things system matrix and impurity, can be used in various living things systems
The quantitative determination of CYP1A1 enzyme activity.
The specific probe reaction can be used to recombinate in single enzyme, people and animal tissue's preparation solution and various
The quantitative determination of CYP1A1 enzyme activity, it may also be used for the quantitative assessment of the inhibitor and rejection ability of quick screening CYP1A1 enzymes.
Using the mono- enzyme of recombinant C YP1A1, liver microsomes incubation system is investigated, and by correlation analysis, specificity suppresses
Many evidences such as experiment, the single enzyme metabolic response of restructuring, and enzyme kinetics, it was demonstrated that resorufin class compound 7- hydroxyls
Ether derivative can specificity Jing CYP1A1 metabolism, generate corresponding hydrolysate.
Used as the fluorescence probe substrate of the CYP1A1 of high specific, the compound can be used to the activity for detecting CYP1A1,
It is especially suitable for the CYP1A1 weights to the production of bacterium, insect cell, mammalian cell and saccharomycete clonal expression system
CYP1A1 in the enzyme activity determination of group enzyme, and the prepared product such as the tissue microsomal of various mammalian tissues organ origins, S-9
Activity is demarcated.
From CYP1A1 of the present invention Specific probe detection CYP1A1 enzyme external activities have it is following prominent
Advantage：
(1) high specific：The ether derivative of resorufin class compound 7- hydroxyls can be by CYP1A1 metabolism with high specificity
Into a metabolite, the i.e. hydrolysate of 7 ether bond ruptures.
(2) it is cheap and easy to get：The ether derivative and its hydrolysate of resorufin class compound 7- hydroxyls can Jing chemistry conjunctions
Into acquisition, synthesis technique is simple.
(3) high sensitivity：Compound with resorufin mother nucleus structure is respectively provided with good fluorescence emission spectral property
(560～630nm), the substrate and its hydrolysis metabolite have different fluorescence emission spectrum signatures, can preferably carry out area
Go-on-go is surveyed, while can be quantitative determined by drawing calibration curve.
Description of the drawings
The mono- enzyme selectivities of Fig. 1
Fig. 2 7- chloroethoxies resorufins and Cytochrome P450 1A1 concentration linear relationship
Fig. 3 7- chloroethoxies resorufins and Cytochrome P450 1A1 dynamic curve
Fig. 4 7- chloroethoxy resorufin cytotoxicities
Fig. 5 7- chloroethoxy resorufin cell imagings
Specific embodiment
The following examples will be further described to the present invention, not thereby limiting the invention.
The synthesis of 1 7- ethoxyresorufins (EHPO) of embodiment
By 1mmol resorufins, 2mmol potassium carbonate, and 5mmol iodoethane are placed in 10mL two-mouth bottles, add 5mL
DMF, is heated to 100 DEG C under nitrogen protective condition, and reaction is stirred overnight.Room temperature is cooled to, reactant liquor 50mL is added to into cold
In water, 10min is stirred vigorously, separates out a large amount of dark red solids, filtered, filter cake adopts column chromatography separation, and (solvent is acetic acid
Ethyl ester: petroleum ether=1: 1, v: v), obtains 135.7mg dark red solids (yield 56.3%).1H NMR(400MHz,DMSO-
D6) δ 7.77 (1H, d, J=8.9), 7.53 (1H, d, J=9.8), 7.11 (1H, s), 7.05 (1H, d, J=9.0), 6.78 (1H,
D, J=9.9), 6.27 (1H, d, J=1.6), 4.20 (2H, dd, J=13.3,6.4), 1.37 (3H, t, J=6.9).13C NMR
(100MHz,DMSO-d6)δ185.76,163.13,150.21,145.80,145.52,135.37,134.14,131.80,
128.28,114.53,106.08,101.16,65.06,14.84.HRMS(ESI positive)[M+H]+Theoretical value
242.0812, measured value 242.0807.
The synthesis of 2 7- chloroethoxy resorufins (EHPO) of embodiment
By 1mmol resorufins, 2mmol potassium carbonate, and 5mmol iodo chloroethanes are placed in 10mL two-mouth bottles, add 5mL
DMF, is heated to 100 DEG C under nitrogen protective condition, and reaction is stirred overnight.Room temperature is cooled to, reactant liquor 50mL is added to into cold
In water, 10min is stirred vigorously, separates out a large amount of dark red solids, filtered, filter cake adopts column chromatography separation, and (solvent is acetic acid
Ethyl ester: petroleum ether=1: 1, v: v), obtains 75.8mg dark red solids (yield 27.6%).1H NMR(400MHz,DMSO-d6)
δ 7.79 (1H, d, J=8.8), 7.54 (1H, d, J=9.8), 7.17 (1H, s), 7.10 (1H, d, J=8.7), 6.80 (1H, d, J
=9.8), 6.28 (1H, s), 4.44 (2H, m), 4.01 (2H, m).13C NMR(100MHz,DMSO-d6)δ185.81,162.39,
150.17,145.91,145.71,135.41,134.29,131.85,128.58,114.51,106.15,101.49,69.50,
43.23.HRMS(ESI positive)[M+H]+Theoretical value 276.0422, measured value 276.0426.
The synthesis of 3 7- propargyl resorufins (PHPO) of embodiment
By 1mmol resorufins, the 3- propargyl bromides of 2mmol potassium carbonate, and 5mmol are placed in 10mL two-mouth bottles, add 5mL
DMF, is heated to 100 DEG C under nitrogen protective condition, and reaction is stirred overnight.Room temperature is cooled to, reactant liquor 50mL is added to into cold
In water, 10min is stirred vigorously, separates out a large amount of dark red solids, filtered, filter cake adopts column chromatography separation, and (solvent is acetic acid
Ethyl ester: petroleum ether=1: 1, v: v), obtains 58.7mg dark red solids (yield 23.4%).1H NMR(400MHz,DMSO-d6)
δ 7.81 (1H, d, J=8.9), 7.54 (1H, d, J=9.8), 7.17 (1H, d, J=2.6), 7.10 (1H, dd, J=8.9,
2.6), 6.80 (1H, dd, J=9.8,2.0), 6.29 (1H, d, J=2.0), 5.00 (2H, d, J=2.2), 3.70 (1H, s).13C
NMR(100MHz,DMSO-d6)δ185.87,161.44,150.16,146.13,145.49,135.43,134.34,131.76,
128.70,114.57,106.20,101.87,79.69,78.78,56.99.HRMS(ESI positive)[M+H]+Theoretical value
252.0655, measured value 252.0656.
The synthesis of 4 7- benzyl resorufins (PHPO) of embodiment
By 1mmol resorufins, 2mmol potassium carbonate, and 5mmol benzyl iodides are placed in 10mL two-mouth bottles, add 5mL
DMF, is heated to 100 DEG C under nitrogen protective condition, and reaction is stirred overnight.Room temperature is cooled to, reactant liquor 50mL is added to into cold
In water, 10min is stirred vigorously, separates out a large amount of dark red solids, filtered, filter cake adopts column chromatography separation, and (solvent is acetic acid
Ethyl ester: petroleum ether=1: 1, v: v), obtains 170.9mg dark red solids (yield 56.4%).1H NMR(400MHz,DMSO-
D6) δ 7.79 (1H, d, J=8.8), 7.58-7.30 (5H, m), 7.21 (1H, s), 7.14 (1H, d, J=8.7), 6.79 (1H, d,
J=9.6), 6.28 (1H, s), 5.28 (2H, s).13C NMR(100MHz,DMSO-d6)δ185.81,162.78,150.20,
145.75,145.70,136.47,135.40,134.21,131.81,129.05,128.70,128.45,114.80,106.12,
101.67,70.79.HRMS(ESI positive)[M+H]+Theoretical value 304.0968, measured value 304.0969.
Selectivity in the recombinant expressed Cytochrome P450 of embodiment 5 each hypotype
57 μ L of buffer solution of potassium phosphate, G-6-P dehydrogenation are separately added in final volume is 100 μ L incubation systems
10 μ L of enzyme, 10 μ L of G-6-P, MgCl2The mono- enzymes of 10 μ L and CYP (CYP1A1, CYP1A2, CYP1B1, CYP2A6,
CYP2A13、CYP2B6、CYP2A13、CYP2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP2J2、
CYP3A4, CYP3A5, CYP4F2 or CYP4F3) 2 μ L (final concentration 20nM), add 1 μ L of EHPO, CHPO, PHPO or BHPO (eventually
50 μM of concentration), with 10 μ L NADP+Initial action, is incubated 60min altogether, is subsequently adding 100 μ L ice acetonitrile terminating reactions, mixes,
20000 × g is centrifuged 20min, takes 150 μ L of supernatant liquid, is detected using ELIASA.(see Fig. 1)
Embodiment 6 is recombinated the protein concentration of single enzyme CYP1A1 linear responses
Buffer solution of potassium phosphate, glucose-6-phosphate dehydrogenase (G6PD) 20 are separately added in final volume is 200 μ L incubation systems
μ L, 20 μ L of G-6-P, MgCl2(final concentration is distinguished for 20 2 μ L (final concentration of 1 μM) of μ L and CHPO, 2 μ L of CYP1A1
For 0,0.15,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9nM, be diluted to respective concentration with buffer solution of potassium phosphate), use
20 μ L NADP+ initial actions.Reaction time 10min, adds 200 μ L acetonitrile terminating reactions, mixes, 20000 × g centrifugations
20min, is taken 150 μ L of supernatant liquid, is detected using ELIASA.(see Fig. 2).Linearity curve equation is Y=2027*X-276 (R2
=0.9960), wherein Y represents fluorescence intensity, and X represents Cytochrome P450 1A1 concentration.
The Chemical Inhibition experiment of 7 probe CHPO of embodiment
116 μ L of buffer solution of potassium phosphate are separately added in final volume is 200 μ L incubation systems, G-6-P takes off
20 μ L of hydrogen enzyme, 20 μ L of G-6-P, MgCl220 μ L, P450 enzyme selectivities inhibitor and its concentration are distinguished as follows：White lamb's-quarters
Reed alcohol (resveratrol, 25 μM, CYP1A specific inhibitors), α-naphthoflavene (ANF, 1 μM, CYP1A specific inhibitors),
Montelukast (5 μM, montelukast, CYP2C8 specific inhibitor), sulfaphenazolum (10 μM, sulfaphenazole,
CYP2C9 specific inhibitors), Omeprazole (omeprazole, 20 μM, CYP2C19 specific inhibitors), quinindium
(quinidine, 10 μM, CYP2D6 specific inhibitors), (clomethiazole, 50 μM, CYP2E1 is specific for clormethiazole
Inhibitor), ketoconazole (ketoconazole, 1 μM, CYP3A4 specific inhibitors), parnitene (2.5 μM,
Tranylcypromine, CYP2A6 specific inhibitor), triethylene thiophosphoramide (50 μM,
Triethylenephosphor amide, TEPA, CYP2B6 specific inhibitor), troleandomycin (troleandomycin,
2.5 μM, CYP3A specific inhibitors) and wide spectrum P450s enzyme inhibitor amino BTAs (1-aminobenzotriazole,
ABT, 500 μM), 1 μ L of CHPO compounds (40 μM of final concentration), 2 μ L of people's hepatomicrosome (final concentration of 0.2 μ g/mL), with 20 μ L
NADP+Initial action, suppresses the percentage that fraction is reduced by fluorescence intensity level to represent.
Embodiment 8 recombinate single enzyme CYP1A1 linear responses time dynamics determine
Buffer solution of potassium phosphate, glucose-6-phosphate dehydrogenase (G6PD) 20 are separately added in final volume is 200 μ L incubation systems
20 μ L and CHPO2 μ L (final concentration of 1 μM) of μ L, 20 μ L of G-6-P, MgCl2,2 μ L of CYP1A1 (are buffered with potassium phosphate
Solution dilutes 5 times, final concentration of 2nM), with 20 μ L NADP+ initial actions.Reaction time be respectively 0,3,6,9,12,15,18,
21st, 24,27min, adds 200 μ L acetonitrile terminating reactions, mixes, 20000 × g centrifugation 20min, takes 150 μ L of supernatant liquid, using enzyme
Mark instrument is detected.(see Fig. 3)
9 probe CHPO cytotoxicities of embodiment are detected
The cytotoxicity of probe CHPO is determined using Sulfonyl rhodamine-B assay (Sulforhodamine B, SRB).At selection
In the A549 cells of exponential phase, the probe of variable concentrations is added in culture medium, after incubation 48h, by three chloroethenes of ice
Sour (10%, w/v, 100 μ L) is added in adherent cell, is kept 1h at 4 DEG C, is subsequently adding SRB solution (final concentration 0.5mg/
L), washed with 1% acetic acid solution (v/v) after 30min being incubated at 37 DEG C, be subsequently adding 100 μ L tris-HCl cushioning liquid, survey
The absorbance being scheduled at 570nm.(see Fig. 4)
The activity of CYP1A1 in the quantitative determination human pulmonary epithelial cells of embodiment 10
A549 cells MEM/EBSS cultivates base (containing 10%FBS) and cultivates, when cell covers with 70% blake bottle, at digestion
Reason, and the incubator containing 5%CO2,37 DEG C of incubated overnights are placed in the concentration plantation in 1 × 105/hole in copolymerization Jiao's capsule.
Culture medium of the attached cell without FBS, adds 2.5 μM of CHPO of probe to continue culture 2h at 37 DEG C, wherein suppressing after rinsing 2 times
Group adds inhibitor Resveratrol (10 μM) preincubate 30min to add probe culture in advance.Cell Jing PBS are rinsed 3 times
Afterwards, it is placed under Laser Scanning Confocal Microscope and observes.Excitation wavelength 543nm, fluorescent collecting scope 570-620nm (see Fig. 5).
Claims (5)
1. the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1 (CYP1A1), it is characterised in that：It is described
The ehter bond of probe substrate can be corresponding hydrolysate by CYP1A1 specific for hydrolysis and produce the corresponding fluorescence of product；By quantitative
In the detection unit interval, the growing amount of hydrolysate is determining the activity of CYP1A1 in different enzyme sources；
Formula（1）
The probe substrate be resorufin ether derivative, its general structure such as formula（1）Shown, wherein R is methyl, ethyl, chlorine
Ethyl, n-propyl, normal-butyl, n-pentyl, n-hexyl, phenyl, p-methylphenyl, to ethylphenyl, to propyl group phenyl, to hexyl
Propyl group, p-nitrophenyl, rubigan, 1- naphthyls, 2- furyls, 2- thienyls,（4- phenyl）Phenyl, 4- ethoxyl phenenyls take
One kind of Dai Jizhong.
2. the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1 according to claim 1, which is special
Levy and be：The enzyme source is human or animal tissues preparation solution, the mono- enzymes of recombinant expressed CYP1A1 or various biology sample
Product.
3. the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1 according to claim 1, which is special
Levy and be：The hydrolysis reaction system pH is between 5.5 ~ 10.5；The concentration of probe substrate is between 1/10 ~ 10K mIt
Between；Incubation system reaction temperature is between 20 ~ 60oBetween C, at the same the conversion ratio of hydrolysate should between 0.1% ~ 20% it
Between.
4. the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1 according to claim 1, which is special
Levy and be：The probe substrate does not possess fluorescence and its hydrolysate has fluorescence properties, can realize producing using fluorescence detector
The rapid sensitive detection of thing and substrate；Fluoroscopic examination condition is：480 ~ 530 nm of excitation wavelength, is carried out in 560 ~ 630 nm
The detection of fluorescence emission spectrum.
5. the application of the high specific fluorescence probe of class detection Cytochrome P450 1A1 according to claim 1, which is special
Levy and be：The probe substrate and its hydrolysis can be used for the quick screening of CYP1A1 inhibitor and quantitatively commenting for rejection ability
Valency.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611115346.1A CN106544399B (en) | 2016-12-07 | 2016-12-07 | Application of high-specificity fluorescent probe for detecting cytochrome P4501A1 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611115346.1A CN106544399B (en) | 2016-12-07 | 2016-12-07 | Application of high-specificity fluorescent probe for detecting cytochrome P4501A1 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106544399A true CN106544399A (en) | 2017-03-29 |
| CN106544399B CN106544399B (en) | 2020-04-03 |
Family
ID=58396573
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611115346.1A Active CN106544399B (en) | 2016-12-07 | 2016-12-07 | Application of high-specificity fluorescent probe for detecting cytochrome P4501A1 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106544399B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113720813A (en) * | 2020-06-08 | 2021-11-30 | 徐州医科大学 | Near-infrared fluorescent probe for detecting cytochrome P4502C 9 and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6143492A (en) * | 1998-12-14 | 2000-11-07 | Aurora Biosciences Corporation | Optical molecular sensors for cytochrome P450 activity |
-
2016
- 2016-12-07 CN CN201611115346.1A patent/CN106544399B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6143492A (en) * | 1998-12-14 | 2000-11-07 | Aurora Biosciences Corporation | Optical molecular sensors for cytochrome P450 activity |
Non-Patent Citations (3)
| Title |
|---|
| ZI-RU DAI ET AL.: "A Highly Selective Ratiometric Two-Photon Fluorescent Probe for Human Cytochrome P450 1A", 《JOURNAL OF THE AMERICAN CHEMICAL SOCIETY》 * |
| 张翠丽: "大蒜油对二乙基亚硝胺诱发肝癌的预防作用及机制研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 李照等: "试卤灵类光学探针的研究进展", 《影像科学与光化学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113720813A (en) * | 2020-06-08 | 2021-11-30 | 徐州医科大学 | Near-infrared fluorescent probe for detecting cytochrome P4502C 9 and application thereof |
| CN113720813B (en) * | 2020-06-08 | 2022-07-19 | 徐州医科大学 | Near-infrared fluorescent probe for detecting cytochrome P4502C 9 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106544399B (en) | 2020-04-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105219374B (en) | Cytochrome oxidase CYP1A Ratiometric fluorescent probe substrate and its application | |
| Tolosa et al. | Neonatal livers: a source for the isolation of good-performing hepatocytes for cell transplantation | |
| Grkovic et al. | National Cancer Institute (NCI) program for natural products discovery: Rapid isolation and identification of biologically active natural products from the NCI prefractionated library | |
| CN110746410B (en) | Leucine aminopeptidase and monoamine oxidase activated near-infrared fluorescent probe, synthetic method and biological application | |
| CN105884734B (en) | A kind of two-photon fluorescence probe of quick detection nitroreductase | |
| Marc et al. | New N-(oxazolylmethyl)-thiazolidinedione active against Candida albicans biofilm: potential Als proteins inhibitors | |
| Marc et al. | Design, synthesis and biological evaluation of new piperazin-4-yl-(acetyl-thiazolidine-2, 4-dione) norfloxacin analogues as antimicrobial agents | |
| CN107022349A (en) | Cytochrome oxidase CYP1A1 specificity fluorescent probes and preparation method and application | |
| CN109928927A (en) | Application of two-photon type fluorescent probe for detecting cytochrome oxidase CYP3A4 | |
| Liu et al. | Structure based discovery of novel hexokinase 2 inhibitors | |
| CN106749153A (en) | The specificity fluorescent probe of nitroreductase and its preparation and the application for cancer target fluorescence imaging and monitoring tumor hypoxia degree | |
| Scalese et al. | Heteroleptic Oxidovanadium (V) complexes with activity against infective and non-infective stages of Trypanosoma cruzi | |
| kumar Marvadi et al. | Synthesis, in vitro, and in vivo (Zebra fish) antitubercular activity of 7, 8-dihydroquinolin-5 (6H)-ylidenehydrazinecarbothioamides | |
| Huang et al. | Elevated hypochlorous acid levels in asthmatic mice were disclosed by a near-infrared fluorescence probe | |
| Dai et al. | In situ target enzyme-activated near-infrared fluorescent probe: a case study of CYP2J2 using three-fragmentary molecular assembly engineering | |
| Shi et al. | The cocultured Nigrospora oryzae and Collectotrichum gloeosporioides, Irpex lacteus, and the plant host Dendrobium officinale bidirectionally regulate the production of phytotoxins by anti-phytopathogenic metabolites | |
| Carter et al. | Natural-product-inspired compounds as countermeasures against the liver carcinogen aflatoxin B1 | |
| Pontecorvi et al. | Novel insights on human carbonic anhydrase inhibitors based on coumalic acid: design, synthesis, molecular modeling investigation, and biological studies | |
| CN106544399A (en) | The application of the high specific fluorescence probe of one class detection Cytochrome P450 1A1 | |
| Casertano et al. | Identifying human PTP1B enzyme inhibitors from marine natural products: perspectives for developing of novel insulin-mimetic drugs | |
| Shi et al. | Endoplasmic Reticulum-Targeting Two-Photon Fluorescent Probe for CYP1A Activity and Its Imaging Application in Endoplasmic Reticulum Stress | |
| Etzerodt et al. | Elucidating the transformation pattern of the cereal allelochemical 6-methoxy-2-benzoxazolinone (MBOA) and the trideuteriomethoxy analogue [D3]-MBOA in soil | |
| Hou et al. | Culture and establishment of self-renewing human liver 3D organoids with high uric acid for screening antihyperuricemic functional compounds | |
| Bon et al. | Synthesis and biological activity of a cytostatic inhibitor of MLLr leukemia targeting the DOT1L protein | |
| CN104910074B (en) | One class contains pyrazole derivatives of hydroxamic acid group as well as preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230403 Address after: 201102 room 801, No. 56, Lane 1266, Gudai Road, Minhang District, Shanghai Patentee after: Wang Zheng Address before: No. 6, Anmin Road, Huangdai Town, Xiangcheng District, Suzhou City, Jiangsu Province, 215131 Patentee before: SUZHOU SHANGJI ELECTRONIC TECHNOLOGY Co.,Ltd. |