CN108588167A - A kind of medical compounds high-throughput screening method - Google Patents
A kind of medical compounds high-throughput screening method Download PDFInfo
- Publication number
- CN108588167A CN108588167A CN201810381029.7A CN201810381029A CN108588167A CN 108588167 A CN108588167 A CN 108588167A CN 201810381029 A CN201810381029 A CN 201810381029A CN 108588167 A CN108588167 A CN 108588167A
- Authority
- CN
- China
- Prior art keywords
- compound
- added
- nadph
- medical compounds
- throughput screening
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5067—Liver cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Cell Biology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to drug screening technology fields, disclose a kind of medical compounds high-throughput screening method.Metabolic enzyme source and its assisted reaction factor is added in the present invention in traditional medicament high flux screening active ingredients incubation system, environment in aids drug body, both drug effects with target effect are filtered out according to activity suppression curve while there is the preferable medical compounds of metabolic stability, the compound filtered out has higher druggability, it is also possible to find active metabolite simultaneously, and then finds new candidate compound.
Description
Technical field
The present invention relates to drug screening technology field, more particularly to a kind of medical compounds high-throughput screening method.
Background technology
Original new drug screening needs pharmaceutical chemistry, pharmacology, drug metabolism, toxicology and preparation etc. are multidisciplinary to match jointly
It closes.Current original new drug from the beginning research and develop often several sequential progress of subject, pharmaceutical chemistry application combinatorial chemistry technique according to
Target construction design synthesizes a large amount of compounds, and pharmacology subject application High Throughput Screening Assay carries out active Fast Evaluation and then sieves
Symptom of a trend compound, drug metabolism and toxicology are selected using early stage drug metabolism and finds that the cooperation progress of toxicological techniques platform is high
Flux evaluation further determines that out lead compound, although promoting the speed and success rate of new drug development under this pattern, still
There are some major issues.Since every subjects technology platform is relatively independent, the screening of whole wheel take it is longer, need chemical combination object amount compared with
Greatly, there are resources and time to waste, and the time is not only money during new drug development, it is often more important that is only quickly
Patient provides medicine, rescue slight illness is only the objective of new drug development.
High flux screening is the important model that current original new drug is found, but high flux screening normally only considers that activity is commented
Valence, the i.e. non-metabolic activity system such as enzyme, receptor, ion channel, nucleic acid and cell of application expression are evaluated as screening model
Compound is to specific actions such as combination, the inhibition of known target, as the starting point of drug screening, although the compound tool filtered out
There is target to combine activity, but far can not also become drug.Determine that can compound not only be decided by its activity as drug,
Drug metabolism stability is also most important.Metabolic stability can determine drug bioavailability, half-life period, therefore metabolic stability
It is to determine that can compound become one of the important feature of drug.
Current conventional medicament high flux screening is generally in 96-384 orifice plates by screening compounds and target receptor, enzyme etc.
Be incubated simultaneously, in incubation system be added isotope labelling competitive inhibitor, or be incubated after using addition fluorescent dye,
The methods of fluorescent reporter protein carries out high-throughput radioactivity or fluoroscopic examination to compound and enzyme conjugates, quickly determines chemical combination
Whether object inhibits with target or activation screens drug activity, does not consider that drug is all in vivo be present in generation in environment
The compound thanked in active environment, therefore evaluated is had a greatly reduced quality as the success rate of drug.
Invention content
The present invention is to overcome the problems, such as that traditional high flux screening is concerned only with target activity, and it is high to provide a kind of medical compounds
Thoroughput screening method, the compound that this method filters out have the double dominant of pharmacological activity and metabolic stability, will carry significantly
The success rate of high medicament high flux screening.
Technical scheme is as follows:
Medical compounds high-throughput screening method of the present invention, includes the following steps:
High flux screening is added in compound to be screened to be incubated in incubation system, the incubation system includes
Two groups for being added and being not added with NADPH response factors are arranged in parallel in specific target receptor, hepatomicrosome or liver S9;Measurementization
It closes object be added without NADPH and be added in the incubation system of NADPH after incubation to the inhibitory activity of target receptor, obtains two work
Property suppression curve;
Compound to be screened is judged according to the activity suppression curve, if two activity suppression curve co-insides, compound
Stability and druggability it is good;If relative to the activity suppression curve that NADPH systems are not added, add the activity suppression of NADPH systems
Koji-making line moves to right, then the stability of compound and druggability are poor;If relative to the activity suppression curve that NADPH systems are not added,
Add the activity suppression curve of NADPH systems to move to left, then prompts compound in vivo can be through the Viability metabolin of metabolism activation.
Preferably, the specific target receptor is opiate receptor or glucagon receptor.
Preferably, the compound to be screened, which is added after being serially diluted in incubation system, is incubated.
It is furthermore preferred that the compound to be screened in incubation system a concentration of 10-12~10-2mol/L。
Preferably, a concentration of 0.2~0.5mg/ml of the hepatomicrosome in incubation system, the liver S9 are being incubated body
A concentration of 1~the 2mM of a concentration of 0.5~1.0mg/ml in system, the NADPH in incubation system.
Preferably, the incubation system 3~10min of first preincubate before NADPH is added, is further continued for incubating after NADPH is added
Educate 10~60min.
Preferably, the temperature of the incubation is 36~38 DEG C.
The prior art is compared, and the present invention has the following advantages:
Medical compounds high-throughput screening method of the present invention adds in traditional medicament high flux screening active ingredients incubation system
Enter hepatomicrosome/liver S9 and NADPH.Contain main I phases drug metabolic enzyme in hepatomicrosome/liver S9, it is anti-by redox
Drug should be carried out to metabolic conversion, NADPH is important the confactor for starting redox reaction.Hepatomicrosome/liver S9 and
NADPH is as metabolic activity system, drug effect environment in analogue body, so that the compound that a wheel filters out both is had and makees to target
Activity has preferable metabolic stability simultaneously, eliminates without activity and metabolism unstable compound, greatly improves screening
Go out the druggability of compound, while being also possible to find active metabolite, find new candidate compound, is to find noval chemical compound
One of important channel.
Description of the drawings
Fig. 1 is several the selection results that screening technique of the present invention is likely to occur;
Fig. 2~6 are respectively compound ZBH1# to be screened, 2#, 3# in embodiment 1, and 4#, 5# are being added and are being added without drug
Two suppression curves of the inhibition opioid receptor activity obtained when metabolic enzyme.
Specific implementation mode
The present invention provides a kind of medical compounds high-throughput screening methods, it is intended to by drug effect target screening active ingredients and metabolism
Stability screening combines, and this approach includes the following steps:
High flux screening is added in compound to be screened to be incubated in incubation system, the incubation system includes
Two groups for being added and being not added with NADPH response factors are arranged in parallel in specific target receptor, hepatomicrosome or liver S9;Measurementization
It closes object be added without NADPH and be added in the incubation system of NADPH after incubation to the inhibitory activity of target receptor or enzyme, obtains two
Activity suppression curve;
Compound to be screened is judged according to the activity suppression curve, if two activity suppression curve co-insides, compound
Stability and druggability it is good;If relative to the activity suppression curve that NADPH systems are not added, add the activity suppression of NADPH systems
Koji-making line moves to right, then the stability of compound and druggability are poor;If relative to the activity suppression curve that NADPH systems are not added,
Add the activity suppression curve of NADPH systems to move to left, then prompts compound in vivo can be through the Viability metabolin of metabolism activation.
Compound to be screened of the present invention preferably sieves the diluted chemical compound for series concentration in screening
Choosing, the concentration after the dilution is preferably 10-12~10-2Mol/L, more preferably 10-10~10-4mol/L.The present invention is in principle
The classification of drug is not limited, all medical compounds can method using the present invention screened.
The type of specific target receptor is not particularly limited in the present invention, those skilled in the art can be according to research
The type of the property setting target receptor of target compound, such as opiate receptor or glucagon receptor.Opiate receptor is needle
The target receptor of the drugs such as, analgesia calm to research and development, drug rehabilitation.Target receptor of the present invention can be with chemical combination to be screened
Object reacts, and the effect reacted with target receptor according to compound screens to obtain has certain active pharmaceutical compound to target receptor
Object.
The present invention is also added into hepatomicrosome or liver S9 in incubation system, is preferably people's hepatomicrosome or people liver S9.People
Contain drug metabolic enzyme in hepatomicrosome and people liver S9, can simulated in vivo environment to drug carry out metabolic conversion.It is of the present invention
Hepatomicrosome is preferably 0.2~0.5mg/ml, more preferably 0.3~0.4mg/ml.The concentration of liver S9 is preferably 0.5~1.0mg/
Ml, more preferably 0.8~1.0mg/ml.
The present invention, which is arranged in parallel, is added and is not added the incubation system of NADPH, when adding NADPH in incubation system
When, a concentration of 1~2mMs of the preferably described NADPH in incubation system.Drug metabolism enzymatic activity is activated after NADPH is added,
There is detection compound to be combined with target receptor or inhibiting effect and metabolic stability respond simultaneously in incubation system.
In the present invention, the compound to be screened is incubated in the incubation system of high flux screening, is preferably incubated
Temperature is 36~38 DEG C, more preferably 37 DEG C;The incubation system preferred elder generation 3~10min of preincubate before NADPH is added, more
It is preferred that 5~8min of preincubate, 10~60min of incubation, more preferably 30~40min are further continued for after NADPH is added.
Various concentration drug competition is measured after incubation, in conjunction with the ability of labelled with radioisotope substrate and target receptor
It is fitted with mapping software Prism and draws competitive activity suppression curve.The preparation method of activity suppression curve of the present invention is adopted
With method well-known to those skilled in the art, specially using medicine series log concentration value as abscissa line, corresponding competition
Binding ability (%) is the longitudinal axis, is fitted suppression curve.
The two groups of activity suppression curves obtained according to not being added and being added NADPH, judge pharmaceutical compound according to suppression curve
The target of object combines activity and metabolic stability.
Assuming that five compounds of ABCDE activity in high-throughput screening active ingredients quite, has the rejection ability of same level, but
If the metabolic activity system that hepatomicrosome/liver S9 and NADPH is added is screened simultaneously, it may occur in which following several results (see Fig. 1):
1) the target association reaction of compound A is not affected because metabolic enzyme source is added, two curve co-insides, A chemical combination
The druggability of object is best;
2) compound B has certain metabolic stability sex chromosome mosaicism, because drug prototype degradation actual concentrations decrease, occurs
The slight of activity suppression curve moves to right;And so on, the druggability ratio B of compound C is poor, and the druggability of compound D is worst;
3) moving to left occurs in the suppression curve that NADPH is added in compound E, and prompts compound E in vivo can be through metabolism activation
Viability metabolin is even stronger than drug prototype to the rejection ability of target.
For the active metabolite that compound E is generated, increases metabolism zymoprotein concentration and compound concentration is metabolized
Product conversion, separation, Structural Identification research can further confirm metabolite, this is also the important channel for finding noval chemical compound.
To make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiment to the present invention into
Row detailed description, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
The screening active ingredients that opiate receptors ligands combine are carried out at the same time metabolic stability screening experiment
Experimental procedure:
1. expressing opiate receptor on Chinese hamster ovary celI:Specific steps are shown in document report:Li Yulei, etc., people recombinate opium sample by
Body Chinese hamster ovary celI stablize expression system foundation, Military Medical Science Institute's proceeding, 2009,33, (5):409-485);
2. cell is handled:Cell is collected, is cracked, Membrane protein extraction:The specific steps are take out be filled 90% cell bottle, abandon
Culture solution is removed, is rinsed twice with 3mlPBS.3ml trypsin solutions are added in every bottle of culture bottle, digest thin under 8-10min after-blow
Born of the same parents collect into 10ml centrifuge tubes, 1500rpm, 4 DEG C, centrifuge 8min.Supernatant is removed, PBS buffer solution is added and is incubated 30min.Ice
Bath played syringe needle (No. 4 syringe needles, 0.45 × 13mm disposable syringes) 5-10 times.The lysis of 2/3 volume is added in centrifuge tube
Buffer (about 7ml), high speed centrifugation (40,000 × g, 4 DEG C, 20min).Supernatant is removed, 50mMTris-HCl1ml, ice is added
Bath played syringe needle (No. 4 syringe needles, 0.45 × 13mm disposable syringes) 5 times.2/3 volume is added in centrifuge tube
50mMTris-HCl (pH7.4) (about 7ml), high speed centrifugation (40,000 × g, 4 DEG C, 20min).Supernatant is removed, is added
50mMTris-HCl1ml, ice bath played syringe needle (No. 4 syringe needles, 0.45 × 13mm disposable syringes) 5 times.Add in centrifuge tube
To the 50mMTris-HCl (about 7ml) of 2/3 volume, high speed centrifugation (40,000 × g, 4 DEG C, 20min).Supernatant is removed, will be carried
The albumen taken be dissolved in 1ml reaction buffers (50mMTris-HCl, 1.5mM CaCl2,5mMEDTA, 5mMKCl, 5mMMgCl2,
120mMNaCl.) in (pH7.4).
3. full cell dosage is set to 400,000 in reaction system, with [3H]-Diprenorphine be tagged ligand, setting flag ligand
A concentration of 30nM;
4. waiting for screening compound 5, ZBH1#, 2#, 3#, 4#, (5 of the brand new of designed, designed synthesis are candidates by 5#
Close object), respectively series concentration is diluted to PBS buffer solution.Experiment packet, each compound are divided into two groups, are separately added into cracking
Cell, [3H]-Diprenorphine (final concentration of 30nM), series concentration compound (final concentration of 0,0.04,0.12,0.37,
1.11,3.33,10,30 μM), after 37 DEG C of preincubate 5min of people's hepatomicrosome solution (final concentration of 0.5mg/ml), one of which
NADPH (1mM) is added and starts reaction, another group is added without NADPH, continues after being incubated 60min, adds the Tris punchings of 4 DEG C of precoolings
(Tris50mM is terminated with HCl tune pH value to 7.4) 4ml and is reacted washing lotion.It is filtered through GF/C filter membranes (Whatman), porous cell is received
Storage negative pressure leaching, then 4mlTris flushing liquors is added to rinse 3 times.80 DEG C of filter membrane, which is dried, to be placed in 1ml scintillation solutions overnight,
LS6500 type BECKMAN liquid scintillation instruments measure cpm (countperminute).
Draw activity suppression curve, the results showed that, other than ZBH2#, ZBH3# compound, other compounds do not occur
Competitive binding curve obviously drifts about, and stability is preferable.The competitive binding curve of ZBH3# and receptor occurs after NADPH is added
It moves to right (see Fig. 4), (see Fig. 3), prompting occurs obviously moving to right after NADPH is added in the competitive binding curve of ZBH2# and receptor
Although the compound has receptor-binding activity, metabolic stability poor.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. a kind of medical compounds high-throughput screening method, includes the following steps:
High flux screening is added in compound to be screened to be incubated in incubation system, the incubation system includes special
Property target receptor, hepatomicrosome or liver S9, be arranged in parallel and be added and be not added with two groups of NADPH response factors;Measure compound
It is being added without NADPH and is being added in the incubation system of NADPH after incubation to the inhibitory activity of target receptor, obtaining two activity suppressions
Koji-making line;
Judge compound to be screened according to the activity suppression curve, if two activity suppression curve co-insides, compound it is steady
Qualitative and druggability is good;If relative to the activity suppression curve that NADPH systems are not added, add the activity suppression of NADPH systems bent
Line moves to right, then the stability of compound and druggability are poor;If relative to the activity suppression curve that NADPH systems are not added, add
The activity suppression curve of NADPH systems moves to left, then prompts compound in vivo can be through the Viability metabolin of metabolism activation.
2. medical compounds high-throughput screening method according to claim 1, which is characterized in that it is described specificity target by
Body is opiate receptor or glucagon receptor.
3. medical compounds high-throughput screening method according to claim 1, which is characterized in that the compound to be screened
It is added in incubation system and is incubated after being serially diluted.
4. medical compounds high-throughput screening method according to claim 3, which is characterized in that the compound to be screened
A concentration of 10 in incubation system-12~10-2mol/L。
5. medical compounds high-throughput screening method according to claim 1, which is characterized in that the hepatomicrosome is being incubated
Educate a concentration of 0.2~0.5mg/ml in system.
6. medical compounds high-throughput screening method according to claim 1, which is characterized in that the liver S9 is being incubated body
A concentration of 0.5~1.0mg/ml in system.
7. according to the medical compounds high-throughput screening method described in any one of claim 1 or 5~6, which is characterized in that institute
State a concentration of 1~2mMs of the NADPH in incubation system.
8. medical compounds high-throughput screening method according to claim 1, which is characterized in that the incubation system is adding
Enter 3~10min of first preincubate before NADPH, 10~60min of incubation is further continued for after NADPH is added.
9. the medical compounds high-throughput screening method according to claim 1 or 8, which is characterized in that the temperature of the incubation
Degree is 36~38 DEG C.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810381029.7A CN108588167B (en) | 2018-04-25 | 2018-04-25 | A kind of medical compounds high-throughput screening method |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810381029.7A CN108588167B (en) | 2018-04-25 | 2018-04-25 | A kind of medical compounds high-throughput screening method |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108588167A true CN108588167A (en) | 2018-09-28 |
CN108588167B CN108588167B (en) | 2019-06-11 |
Family
ID=63609743
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810381029.7A Active CN108588167B (en) | 2018-04-25 | 2018-04-25 | A kind of medical compounds high-throughput screening method |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108588167B (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110289055A (en) * | 2019-06-25 | 2019-09-27 | 中国人民解放军军事科学院军事医学研究院 | Prediction technique, device, computer equipment and the storage medium of drug targets |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006026100A2 (en) * | 2004-08-26 | 2006-03-09 | Qualyst, Inc. | Methods and kits for determining metabolic stability of compounds |
CN103937869A (en) * | 2014-04-13 | 2014-07-23 | 中国检验检疫科学研究院 | Kit for screening CYP450 enzyme inhibition medicament with high flux and research method thereof |
CN106405027A (en) * | 2015-07-29 | 2017-02-15 | 上海医药集团股份有限公司 | Method for high throughput determination of in vitro metabolism stability of compound, and applications thereof |
CN106834206A (en) * | 2015-12-07 | 2017-06-13 | 江苏齐氏生物科技有限公司 | A kind of induction of rats'liver S9 and preparation method |
-
2018
- 2018-04-25 CN CN201810381029.7A patent/CN108588167B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2006026100A2 (en) * | 2004-08-26 | 2006-03-09 | Qualyst, Inc. | Methods and kits for determining metabolic stability of compounds |
CN103937869A (en) * | 2014-04-13 | 2014-07-23 | 中国检验检疫科学研究院 | Kit for screening CYP450 enzyme inhibition medicament with high flux and research method thereof |
CN106405027A (en) * | 2015-07-29 | 2017-02-15 | 上海医药集团股份有限公司 | Method for high throughput determination of in vitro metabolism stability of compound, and applications thereof |
CN106834206A (en) * | 2015-12-07 | 2017-06-13 | 江苏齐氏生物科技有限公司 | A kind of induction of rats'liver S9 and preparation method |
Non-Patent Citations (3)
Title |
---|
ALBERT P.LI等: "Screening for human ADME/Tox drug properties in drug discovery", 《DRUG DISCOVERY TODAY》 * |
SAMANTHA J. RICHARDSON等: "Efficiency in Drug Discovery: Liver S9 Fraction Assay As a Screen for Metabolic Stability", 《DRUG METABOLISM LETTERS》 * |
翁俊等: "药物代谢稳定性筛选的研究进展", 《国外医学药学分册》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110289055A (en) * | 2019-06-25 | 2019-09-27 | 中国人民解放军军事科学院军事医学研究院 | Prediction technique, device, computer equipment and the storage medium of drug targets |
CN110289055B (en) * | 2019-06-25 | 2021-09-07 | 中国人民解放军军事科学院军事医学研究院 | Method and device for predicting drug target, computer equipment and storage medium |
Also Published As
Publication number | Publication date |
---|---|
CN108588167B (en) | 2019-06-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Giustino et al. | Coronavirus and cardiovascular disease, myocardial injury, and arrhythmia: JACC focus seminar | |
Ding et al. | Emerging new concepts of degrader technologies | |
Kapoor et al. | Identification of rodent homologs of hepatitis C virus and pegiviruses | |
Stramer et al. | Nucleic acid testing to detect HBV infection in blood donors | |
Weber et al. | Absorption, excretion, and metabolism of the endothelin receptor antagonist bosentan in healthy male subjects | |
Uddayasankar et al. | The pharmacokinetics and pharmacodynamics of carfentanil after recreational exposure: a case report | |
Anwaar et al. | Combined deep learning and molecular docking simulations approach identifies potentially effective FDA approved drugs for repurposing against SARS-CoV-2 | |
Li et al. | Physiologically based pharmacokinetic prediction of telmisartan in human | |
Capitano et al. | Intrapulmonary penetration of voriconazole in patients receiving an oral prophylactic regimen | |
Ton et al. | Targeting SARS-CoV-2 papain-like protease in the postvaccine era | |
Barker Jr et al. | Aminoindoles, a novel scaffold with potent activity against Plasmodium falciparum | |
Eckert et al. | Pharmacokinetics and biotransformation of 2-[N-[(S)-1-ethoxycarbonyl-3-phenylpropyl]-L-alanyl]-(1S, 3S, 5S)-2-azabicyclo [3.3. 0] octane-3-carboxylic acid (Hoe 498) in rat, dog and man. | |
CN104749009A (en) | Immunosuppressant medicine extracting reagent for immunoassay | |
Tran et al. | Titers of antibodies against ancestral SARS-CoV-2 correlate with levels of neutralizing antibodies to multiple variants | |
Chen et al. | Activity of a potent hepatitis C virus polymerase inhibitor in the chimpanzee model | |
CN108588167B (en) | A kind of medical compounds high-throughput screening method | |
Font et al. | Determination of zidovudine triphosphate intracellular concentrations in peripheral blood mononuclear cells from human immunodeficiency virus-infected individuals by tandem mass spectrometry | |
Igloi et al. | Requirement for chloride channel function during the hepatitis C virus life cycle | |
Rocamora et al. | PfMFR3: a multidrug-resistant modulator in Plasmodium falciparum | |
Procko | Deep mutagenesis in the study of COVID-19: a technical overview for the proteomics community | |
Peng et al. | Nonproductive hepatitis B virus covalently closed circular DNA generates HBx-related transcripts from the HBx/Enhancer I region and acquires reactivation by superinfection in single cells | |
Souto et al. | further evidence for hepatitis E in the Brazilian Amazon. | |
Gudima et al. | Primary human hepatocytes are susceptible to infection by hepatitis delta virus assembled with envelope proteins of woodchuck hepatitis virus | |
Wu et al. | Mass spectrometry-based phosphoproteomics in clinical applications | |
Joyamma et al. | Biochemical mechanisms and effects of Mimosa pudica (Linn) on experimental urolithiasis in rats. |
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 |