CN103217463A - Electrochemistry method for screening BACE1 (beta-secretase) inhibitor - Google Patents
Electrochemistry method for screening BACE1 (beta-secretase) inhibitor Download PDFInfo
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Abstract
The invention discloses an electrochemistry method for screening BACE1 (beta-secretase) inhibitor, and the method comprises the following steps: assembling polypeptide molecules labelled by biotin on a gold electrode surface for enclosing the surface of the gold electrode; inhibiting digestion reaction of the gold electrode with enclosed surface in the condition that the BACE 1 inhibitor to be screened and the BACE 1 are existing; deriving mercapto ferrocene modified nano-gold/avidin compound to the surface of the obtained gold electrode after finishing inhibition of the digestion reaction, and screening the BACE 1 inhibitor using the obtained gold electrode by an electrochemistry detection; the method has the advantages of strong response, high test sensitivity, simple step, easy and fast operation, and simultaneously the method can be used for screening a plurality of BACE 1 inhibitors with wide application.
Description
Technical field
The present invention relates to a kind of electrochemical method of the BACE1 of being used for inhibitor screening, belong to the drug screening field.
Background technology
Alzheimer disease (Alzheimer ' s Disease, AD) be a kind of common nerve degenerative diseases, the senile plaque expelling that amyloid-beta (β-amyloid Protein, A β) deposition forms in patient's brain is its main pathological characters.At present major part studies show that the substantial connection that exists of the unconventionality expression of A β and AD.Therefore, become the main means of prevention and treatment AD from the generation of source minimizing A β.
A β is that (Amyloid Precursor Protein is APP) through β-cutting forms with gamma-secretase by transmembrane protein beta amyloid precursor protein.Therefore, the biologically active that suppresses beta-secretase can reduce generation and the gathering of A β subsequently of A β to a certain extent, thereby prevention effect is played in generation, the development of AD.Because APP is a transmembrane protein that molecular weight is bigger, the target polypeptides sequence of employing short chain is simulated beta-secretase the cutting behavior of APP and the inhibitor of screening inhibition beta-secretase activity is had important Research Significance.
Therefore, seeking the medicine that effectively suppresses the beta-secretase activity is one of current research focus.Inhibitor mainly is to screen from the synthetic or natural drug molecule of kinds of artificial at present, although experiment in vitro shows these molecules and can interact with beta-secretase, the activity of inhibitory enzyme does not show that these molecules can be used for the treatment of AD but have clinical trial at present.In addition, the problem that above-mentioned inhibitor exists is that molecular dimension is bigger, and lipophilicity is relatively poor, is difficult to penetrate blood brain barrier.At these problems, people tend to seek the activity that some drug molecules are used to suppress beta-secretase from Drug Storage or natural products.Reported method as FRET (fluorescence resonance energy transfer) (FRET), mainly is to screen inhibitor by medicine or parent molecule (fragment among the APP) are carried out fluorescence labeling.But the polypeptide of fluorophor mark synthesizes more complicated, and the size of labelling groups can influence the polypeptide configuration; In addition, adopt fluorescence labeling to screen that the method step of inhibitor is loaded down with trivial details, the cycle is long, and photobleaching, less stable easily take place in fluorophor, therefore very difficultly from thousands of kinds of drug molecules, filter out effective enzyme inhibitor.
Summary of the invention
The method step that the present invention is directed to present FRET (fluorescence resonance energy transfer) (FRET) screening beta-secretase inhibitor is loaded down with trivial details, the cycle is long, and photobleaching, less stable easily take place in fluorophor when adopting fluorescence labeling, thereby cause to carry out a series of problems such as effective screening of drug molecule, provide a kind of highly sensitive, step is simple, easy and simple to handle, quick, can screen the electrochemical method of multiple BACE1 inhibitor simultaneously.
The invention provides a kind of electrochemical method of the BACE1 of being used for inhibitor screening, this method is after biotin labeled peptide molecule is assembled in the surface of gold electrode, with the surface sealing of gold electrode; Sealed surperficial gold electrode and under the condition that BACE1 inhibitor to be screened and BACE1 exist, suppressed endonuclease reaction with above-mentioned; Nm of gold/avidin compound that the sulfydryl ferrocene is modified is derived to the surface of the gold electrode that suppresses to obtain after endonuclease reaction is finished again, and carries out the screening of BACE1 inhibitor by Electrochemical Detection with the resulting gold electrode of deriving.
The screening of described BACE1 inhibitor is at least 3 to have been assembled biotin labeled peptide molecule and sealed surperficial gold electrode to place concentration be that the mixed liquor of the BACE1 inhibitor to be screened of the BACE1 of 5～15nM and variable concentrations suppresses endonuclease reaction; Finish the derive nm of gold/avidin compound of sulfydryl ferrocene modification of the surface of the resulting gold electrode in back suppressing endonuclease reaction, carry out Electrochemical Detection again; Determine to suppress the relation of effect and inhibitor concentration by electrochemical response, and use IC
50Value is judged the inhibition effect of BACE1 inhibitor to be screened, and carries out the screening of BACE1 inhibitor.
The screening of described BACE1 inhibitor need be done the inhibitory enzyme of the BACE1 inhibitor to be screened of 3 groups of variable concentrations at least and cut reaction experiment, up to the inhibition effect that can determine BACE1 inhibitor to be screened and the linear relationship of inhibitor concentration.
Described IC
50Reach the concentration of a highest half for the inhibition effect of inhibitor.
The reaction conditions of described inhibition endonuclease reaction is to react 30～90min in 37 ℃ environment.
Described BACE1 pH is 4～5 HAc-NaAc damping fluid dissolving, and the BACE1 inhibitor dissolves with DMSO.
Nm of gold/avidin compound that described sulfydryl ferrocene is modified obtains by the following method: with HS-(CH
2)
6The Fc ultrasonic dissolution adds the nm of gold/avidin compound of PBS dilution in above-mentioned solution in normal hexane, shake 20～28h under the rotating speed of 800～1200rps, centrifugal organic phase and the water of discarding, promptly.
The pH of described PBS is 7～7.5.
Described sealing be will assemble the gold electrode of biotin labeled peptide molecule soak 4～10min with the HT of 0.1mM earlier, again with BSA immersion 0.5～1.5h of 1%.
Described biotin labeled peptide molecule is soluble in water.
Described biotin labeled peptide molecule sequence is that the Bition-KTEEISEVN LDA EFRHDKC(producer such as bio tech ltd that can shine by force in the prosperous Bioisystech Co., Ltd of Sigma-Aldrich, Shanghai bioengineering company limited, Beijing ancient cooking vessel state, Shanghai buys).
Nm of gold/avidin that the present invention adopts is commercially available conventional biological reagent (S9059-2mL can buy the producers such as bio tech ltd that shine by force in Sigma-Aldrich, Shanghai bioengineering company limited, the prosperous Bioisystech Co., Ltd of Beijing ancient cooking vessel state, Shanghai).
Described Electrochemical Detection is that the gold electrode that will assemble places 0.1～0.5M KClO
4Aqueous solution in carry out.
Described BACE1 inhibitor to be screened only with the BACE1 effect, and not with the peptide substrate effect.
The present invention carried out the endonuclease reaction of BACE1 earlier before suppressing endonuclease reaction, conditions such as the concentration when BACE1 is carried out endonuclease reaction, temperature and time preferred; Concentration is preferably 5～15nM; Temperature is preferably 37 ℃; Time is preferably 30～90min.
The concrete preparation method of nm of gold/avidin compound that sulfydryl ferrocene of the present invention is modified is:
(1) pipettes 1.7 μ L HS-(CH with liquid-transfering gun
2)
6Fc(or take by weighing 0.0008g) to 600～800 μ L normal hexanes, ultrasonic dissolution;
(2) adopting PBS(pH is 7～7.5) 100 μ L nm of gold (the about 10nm of diameter)/avidin compound is diluted to 200 μ L;
(3) step (1) and (2) gained solution are mixed, place on the circumference vibration shaking table, under the rotating speed of 800～1200rps, shake 20～28h;
(4) centrifugally discard normal hexane organic phase and unreacted water, thereby obtain nm of gold-avidin compound that ferrocene is modified;
(5) resulting nano-Au composite is with normal hexane drip washing twice, is placed in the refrigerator (0～4 ℃) and preserves standby.
The concrete grammar that the present invention screens the BACE1 inhibitor is:
(1) gold electrode being immersed in 50 μ L concentration is in the biotin labeled polypeptide of 1 μ M (sequence the is Bition-KTEEISEVN LDA EFRHDKC) solution, behind 8～12h, repeatedly washes with redistilled water, uses N
2Dry up;
(2) the HT sealing 4～10min about 50 μ L of gold electrode elder generation employing 0.1mM(consumption that step (1) obtained) rinses N then well with redistilled water
2Dry up;
(3) on the basis of step (2), adopt the about 50 μ L of 1%(consumption again) BSA sealing gold electrode surfaces 0.5～1.5h, rinse N then well with redistilled water
2Dry up;
(4) endonuclease reaction and inhibition endonuclease reaction divide two groups of experiments to carry out successively:
(a) endonuclease reaction:
(I) gold electrode after many (at least 3) above-mentioned steps (3) sealing being immersed in 50 μ L concentration respectively is in the BACE1 solution of 10nM; Place 37 ℃ of environment, the reaction time rinses gold electrode well between 0～120min with redistilled water, N
2Dry up;
(II) nm of gold/avidin compound that 10 μ L ferrocene are modified is added drop-wise to the surface of the every gold electrode that step (I) obtains, and rinses N behind 20～60min well with redistilled water
2Dry up;
(III) at last the gold electrode that assembles is placed 0.1～0.5M KClO
4Carry out Electrochemical Detection in the aqueous solution;
Optimize best endonuclease reaction reaction time 30～90min of BACE1;
(IV) repeat (I)～(III) step, the soak time in (I) is selected in 30～90min, the concentration of BACE1 is between 0～20nM, and other condition is constant;
The best endonuclease reaction desired concn that optimizes BACE1 is 5～15nM;
(b) suppress endonuclease reaction:
(i) BACE1 inhibitor to be screened and the concentration that the gold electrode after many (at least 3) above-mentioned (3) sealings is immersed in variable concentrations is in the mixed liquor of 5～15nM BACE1; Place 37 ℃ environment to react 30～90min, again each gold electrode is rinsed well N with redistilled water
2Dry up;
(ii) nm of gold/avidin compound that 10 μ L ferrocene are modified is added drop-wise to the resulting every gold electrode surfaces of step (i), and reaction 20～60min rinses N then well with redistilled water
2Dry up;
(iii) at last the gold electrode that assembles is placed 0.1～0.5M KClO
4Carry out Electrochemical Detection in the aqueous solution.
The Electrochemical Detection that the present invention adopts is at ambient temperature, adopt three-electrode system, in the inventive method by endonuclease reaction or the gold electrode that finally obtains after suppressing endonuclease reaction be working electrode, the Ag/AgCl electrode is a contrast electrode, the Pt silk is an auxiliary electrode, sweep speed and be 0.1V/s, sweep limit is 0～0.6V.
Beneficial effect of the present invention: it is substrate that the inventor adopts biotin labeled peptide molecule first, and nm of gold/avidin compound of modifying by the sulfydryl ferrocene amplifies electrochemical response, carries out the screening of BACE1 inhibitor; The inventive method adopts stable and the good substrate of dissolubility, and the nm of gold/avidin compound that adopts the sulfydryl ferrocene to modify simultaneously amplifies electrochemical response, has strengthened measurement sensitivity greatly; The inventive method step is simple, and is easy and simple to handle, quick, can screen multiple BACE1 inhibitor simultaneously, really realized the electrochemical method screening of BACE1 inhibitor.
Description of drawings
[Fig. 1] is the experimental principle figure of BACE1 cutting polypeptide Electrochemical Detection and inhibitor screening thereof.
[Fig. 2] cuts the cyclic voltammogram of peptide sequence front and back for the BACE1 enzyme: a is a hac buffer; B is the mixed liquor of 10nM BACE1 and 120nM BACE1 inhibitor 1; C is the mixed liquor of 10nM BACE1 and 6 μ M BACE1 inhibitor 2; D is 4.5 acetate buffer solution for 10nM BACE1(is dissolved in pH); Wherein, sweep speed and be 0.1V/s, supporting electrolyte is 0.1M KClO
4, e is the direction of scanning.
[Fig. 3] cuts the electrochemical response figure of polypeptide different time for the BACE1 enzyme: wherein, sweep speed and be 0.1V/s, supporting electrolyte is 0.1M KClO
4
[Fig. 4] cuts design sketch for the enzyme of the BACE1 of variable concentrations.
[Fig. 5] is the inhibition design sketch of variable concentrations BACE1 inhibitor 1: wherein, the concentration fixed of BACE1 is 10nM.
[Fig. 6] is the inhibition design sketch of variable concentrations BACE1 inhibitor 2: wherein, the concentration fixed of BACE1 is 10nM.
Embodiment
Following examples are to further specify of the present invention, rather than restriction the present invention.
The BACE1 enzyme is cut the influence of time:
(1) will handling 7 clean gold electrodes, to be immersed in 50 μ L concentration respectively be in the biotin labeled polypeptide of 1 μ M (sequence the is Bition-KTEEISEVN LDA EFRHDKC) solution, behind 8～12h, repeatedly washes N with redistilled water
2Dry up;
(2) adopt the HT of 50 μ L, 0.1mM to seal gold electrode surfaces 5min, rinse N then with redistilled water well
2Dry up;
(3) adopt 50 μ L, 1% BSA sealing gold electrode surfaces 1h, rinse N then with redistilled water well
2Dry up;
(4) enzyme of design different time is cut process:
It is in the BACE1 solution of 10nM that gold electrode after the sealing is immersed in 50 μ L concentration respectively; Place 37 ℃ water-bath to react, soak time is respectively 0,10,30,45,60,90, and 120min rinses gold electrode well with redistilled water, N
2Dry up;
(5) nm of gold/avidin compound that 10 μ L ferrocene are modified is added drop-wise to every gold electrode surfaces, and reaction 30min rinses N well with redistilled water
2Dry up;
(6) at last the gold electrode that assembles is put into 5mL, 0.1M KClO
4Carry out Electrochemical Detection in the aqueous solution.
Electrochemical Detection adopts three-electrode system, and the gold electrode that said method obtains is a working electrode, and the Ag/AgCl electrode is a contrast electrode, the Pt silk is an auxiliary electrode, sweeps speed and is 0.1V/s, and sweep limit is 0～0.6V, the result as shown in Figure 3, all Electrochemical Detection are all carried out at ambient temperature.
The enzyme of variable concentrations BACE1 is cut effect:
Cut the step in the influence experiment of time with reference to the BACE1 enzyme, just change different time in the step (4) into different BACE1 concentration, be respectively 0,0.5,1,2,5,10,20nM, the reaction time is 45min, other condition is constant, and the gold electrode that assembles is placed 0.1M KClO
4Carry out Electrochemical Detection in the aqueous solution, as Fig. 4.
Variable concentrations BACE1 inhibitor 1(sequence is KTEEI-Statine-EVNVAEF) the inhibition effect:
(1) will handle clean gold electrode and be immersed in 50 μ L, the biotin labeled polypeptide solution of 1 μ M, behind 8～12h, repeatedly wash N with redistilled water
2Dry up;
(2) adopt the HT of 50 μ L, 0.1mM to seal gold electrode surfaces 5min, rinse N then with redistilled water well
2Dry up;
(3) adopt 50 μ L, 1% BSA sealing gold electrode surfaces 1h, rinse N then with redistilled water well
2Dry up;
(4) inhibitory enzyme is cut process: the gold electrode after 8 sealings is immersed in BACE1 inhibitor 1 and concentration is in the mixed liquor of 10nMBACE1, volume is 50 μ L, and the concentration of BACE1 inhibitor 1 is respectively 0,5,10,25,40,80,120,160nM places 37 ℃ water-bath to react 45min gold electrode, rinse N well with redistilled water
2Dry up;
(5) nm of gold/avidin compound that 10 μ L ferrocene are modified is added drop-wise to gold electrode surfaces, and reaction 30min rinses N then well with redistilled water
2Dry up;
(6) at last the gold electrode that assembles is placed 5mL, 0.1M KClO
4Carry out Electrochemical Detection in the aqueous solution.
Electrochemical Detection adopts three-electrode system, and the gold electrode that said method obtains is a working electrode, and the Ag/AgCl electrode is a contrast electrode, the Pt silk is an auxiliary electrode, sweeps speed and is 0.1V/s, and sweep limit is 0～0.6V, the result as shown in Figure 5, all Electrochemical Detection are all carried out at ambient temperature.
The effect of BACE1 endonuclease reaction time effects and variable concentrations such as embodiment 1;
BACE1 inhibitor 2(Ph-LL-4,5-dehydro-L-CHO) inhibitory enzyme is cut reaction experiment:
The inhibition effect of variable concentrations BACE1 inhibitor 2:
(1) will handle clean gold electrode and be immersed in 50 μ L, the biotin labeled polypeptide solution of 1 μ M, behind 8～12h, repeatedly wash N with redistilled water
2Dry up;
(2) adopt the HT of 50 μ L, 0.1mM to seal gold electrode surfaces 5min, rinse N then with redistilled water well
2Dry up;
(3) adopt 50 μ L, 1% BSA sealing gold electrode surfaces 1h, rinse N then with redistilled water well
2Dry up;
(4) inhibitory enzyme is cut process: the gold electrode after 8 sealings is immersed in BACE1 inhibitor 2 and concentration is in the mixed liquor of 10nMBACE1, volume is 50 μ L, and the concentration of BACE1 inhibitor 2 is respectively 0,0.5,1,2,4,6,10,15 μ M.Place 37 ℃ water-bath to react 45min above-mentioned gold electrode, gold electrode is rinsed well N with redistilled water
2Dry up;
(5) nm of gold/avidin compound that 10 μ L ferrocene are modified is added drop-wise to gold electrode surfaces, and reaction 30min rinses N then well with redistilled water
2Dry up;
(6) at last the gold electrode that assembles is placed 5mL, 0.1M KClO
4Carry out Electrochemical Detection in the aqueous solution.
Electrochemical Detection adopts three-electrode system, and the gold electrode that said method obtains is a working electrode, and the Ag/AgCl electrode is a contrast electrode, and the Pt silk is an auxiliary electrode, sweeps speed and is 0.1V/s, and sweep limit is 0～0.6V, result such as Fig. 6.
Claims (7)
1. an electrochemical method that is used for the BACE1 inhibitor screening is characterized in that, biotin labeled peptide molecule is assembled in the surface of gold electrode after, with the sealing of the surface of gold electrode; Sealed surperficial gold electrode and under the condition that BACE1 inhibitor to be screened and BACE1 exist, suppressed endonuclease reaction with above-mentioned; Nm of gold/avidin compound that the sulfydryl ferrocene is modified is derived to the surface of the gold electrode that suppresses to obtain after endonuclease reaction is finished again, and carries out the screening of BACE1 inhibitor by Electrochemical Detection with the resulting gold electrode of deriving.
2. the method for claim 1, it is characterized in that the screening of described BACE1 inhibitor is at least 3 to have been assembled biotin labeled peptide molecule and sealed surperficial gold electrode to place concentration be that the mixed liquor of the BACE1 inhibitor to be screened of the BACE1 of 5～15nM and variable concentrations suppresses endonuclease reaction; Finish the derive nm of gold/avidin compound of sulfydryl ferrocene modification of the surface of the resulting gold electrode in back suppressing endonuclease reaction, carry out Electrochemical Detection again; Determine to suppress the relation of effect and inhibitor concentration by electrochemical response, and use IC
50Value is judged the inhibition effect of BACE1 inhibitor to be screened, and carries out the screening of BACE1 inhibitor.
3. method as claimed in claim 2 is characterized in that, the reaction conditions of described inhibition endonuclease reaction is to react 30～90min in 37 ℃ environment.
4. method as claimed in claim 1 or 2 is characterized in that, nm of gold/avidin compound that described sulfydryl ferrocene is modified obtains by the following method: with HS-(CH
2)
6The Fc ultrasonic dissolution adds the nm of gold/avidin compound of PBS dilution in above-mentioned solution in normal hexane, shake 20～28h under the rotating speed of 800～1200rps, centrifugal organic phase and the water of discarding, promptly.
5. method as claimed in claim 1 or 2 is characterized in that, described sealing be will assemble the gold electrode of biotin labeled peptide molecule soak 4～10min with the HT of 0.1mM earlier, again with BSA immersion 0.5～1.5h of 1%.
6. method as claimed in claim 5 is characterized in that, described biotin labeled peptide molecule is soluble in water.
7. method as claimed in claim 1 or 2 is characterized in that, described Electrochemical Detection is that the gold electrode that will assemble places 0.1～0.5M KClO
4Aqueous solution in carry out.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104807865A (en) * | 2015-05-08 | 2015-07-29 | 江苏省农业科学院 | Preparation method of electrochemical aptamer sensor applied to myoglobin detection |
| CN104880441A (en) * | 2015-05-14 | 2015-09-02 | 上海皓拓生物技术有限公司 | Screening method and screening system for beta-secretase specific inhibitor |
| CN106093160A (en) * | 2016-06-07 | 2016-11-09 | 中南大学 | A kind of hydroxy apatite-base electrochemical probe construction method and mensuration BACE1 activity and the method for inhibition |
| CN113929748A (en) * | 2020-07-13 | 2022-01-14 | 中国科学技术大学 | Kit for detecting activity of BACE1 enzyme and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101262863A (en) * | 2005-07-08 | 2008-09-10 | 马泰克生物科学公司 | Polyunsaturated fatty acids for treatment of dementia and pre-dementia-related conditions |
-
2013
- 2013-03-14 CN CN2013100811755A patent/CN103217463A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101262863A (en) * | 2005-07-08 | 2008-09-10 | 马泰克生物科学公司 | Polyunsaturated fatty acids for treatment of dementia and pre-dementia-related conditions |
Non-Patent Citations (3)
| Title |
|---|
| 廖国超 等: "β-分泌酶抑制剂的研究进展", 《中国药物化学杂志》 * |
| 赵彬 等: "β分泌酶抑制剂体外筛选模型的构建与验证", 《中国药理学通报》 * |
| 韩洪星: "基于信号放大检测miRNA和BACE1酶切多肽的电化学研究", 《中国优秀硕士学位论文全文数据库（医药卫生科技辑）》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104807865A (en) * | 2015-05-08 | 2015-07-29 | 江苏省农业科学院 | Preparation method of electrochemical aptamer sensor applied to myoglobin detection |
| CN104807865B (en) * | 2015-05-08 | 2017-06-30 | 江苏省农业科学院 | It is applied to the preparation method of the electrochemical aptamer sensor of myoglobins detection |
| CN104880441A (en) * | 2015-05-14 | 2015-09-02 | 上海皓拓生物技术有限公司 | Screening method and screening system for beta-secretase specific inhibitor |
| CN104880441B (en) * | 2015-05-14 | 2017-12-22 | 上海皓拓生物技术有限公司 | The screening technique and its screening system of beta-secretase specific inhibitor |
| CN106093160A (en) * | 2016-06-07 | 2016-11-09 | 中南大学 | A kind of hydroxy apatite-base electrochemical probe construction method and mensuration BACE1 activity and the method for inhibition |
| CN106093160B (en) * | 2016-06-07 | 2018-11-06 | 中南大学 | A kind of method of hydroxy apatite-base electrochemical probe construction method and measurement BACE1 activity and inhibition |
| CN113929748A (en) * | 2020-07-13 | 2022-01-14 | 中国科学技术大学 | Kit for detecting activity of BACE1 enzyme and application |
| CN113929748B (en) * | 2020-07-13 | 2023-10-20 | 中国科学技术大学 | Kit for detecting BACE1 enzyme activity and application thereof |
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Application publication date: 20130724 |