CN111307982A - In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora - Google Patents

In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora Download PDF

Info

Publication number
CN111307982A
CN111307982A CN202010186816.3A CN202010186816A CN111307982A CN 111307982 A CN111307982 A CN 111307982A CN 202010186816 A CN202010186816 A CN 202010186816A CN 111307982 A CN111307982 A CN 111307982A
Authority
CN
China
Prior art keywords
intestinal
effect
strain
platform
drug candidate
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.)
Pending
Application number
CN202010186816.3A
Other languages
Chinese (zh)
Inventor
刘双江
刘畅
周楠
姜成英
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Microbiology of CAS
Original Assignee
Institute of Microbiology of CAS
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Institute of Microbiology of CAS filed Critical Institute of Microbiology of CAS
Priority to CN202010186816.3A priority Critical patent/CN111307982A/en
Publication of CN111307982A publication Critical patent/CN111307982A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/62Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode

Landscapes

  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Electrochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses an in vitro high-flux screening platform for the effect of a drug candidate on intestinal flora, which is characterized in that a plurality of representative intestinal strains are selected according to the characteristics of the drug candidate and the intestinal flora of target people to simulate a simplified human intestinal microbial environment; the method comprises the steps of diluting the intestinal canal strain in a microbial culture medium, placing the diluted intestinal canal strain in an integrated pore plate, keeping the initial concentration of a bacterial suspension of the intestinal canal strain between 0.2 and 1.0, and realizing high-throughput screening of the action effect of a target drug candidate and the intestinal canal strain according to growth phenotype detection of the intestinal canal strain and microbial metabolite detection of a target compound.

Description

In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora
Technical Field
The invention relates to the technical field of biological detection, in particular to an in-vitro high-throughput screening platform for the effect of a drug candidate on intestinal flora.
Background
In past studies, in vitro screening methods based on in vitro cell culture and target-based drug activity screening (including enzymatic targets, GPCR targets, ion channel targets, nuclear receptor targets, etc.) have been commonly used for new drugs and drug candidates. However, in recent years, with the progress of studies on intestinal microorganisms, it has been found that a considerable part of drugs act on the body indirectly by interacting with the intestinal flora of the human body, in addition to directly interacting with the host by being absorbed into the blood or acting on the target of host cells. In recent researches, two groups in succession respectively prove that the intestinal flora participates in the metabolism and the efficacy of various medicines in Nature and Science journal texts. At present, animal models are mostly adopted to evaluate the interaction between the drug and the intestinal flora, and a high-throughput rapid screening method similar to an in vitro cell screening platform is lacked. Therefore, we developed here an in vitro high throughput screening platform for drug-gut flora effect based on pure culture system of gut strains.
Disclosure of Invention
The invention aims to provide an in-vitro high-throughput screening platform for the effect of a drug candidate on intestinal flora, which aims to solve the existing problems.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention comprises the following steps:
A. selecting a plurality of representative intestinal strains to simulate a simplified human intestinal microbial environment according to the drug candidates and the characteristics of the intestinal flora of the target population;
b, diluting the intestinal tract strains in a microbial culture medium, and then placing the diluted intestinal tract strains in an integrated pore plate, wherein the initial concentration of the bacterial suspension of the intestinal tract strains is kept between 0.2 and 1.0;
C. respectively dripping the target drug candidates into the integrated pore plate through a microfluidic high-flux liquid distribution system, and then carrying out anaerobic culture on the intestinal strain in a continuous gas supply device;
D. culturing for 12-72 hours, and performing growth phenotype detection and metabolite detection based on optical parameters on the intestinal bacteria;
E. and according to the growth phenotype detection of the intestinal bacteria and the microbial metabolite detection of the target compound, the interaction effect of the drug candidate and the intestinal microorganisms is screened at high flux for the interaction effect of the target drug candidate and the intestinal bacteria.
Further, the metabolite detection reflects the effect of different drug candidates on the gut species metabolite detection activity through the gut species biomass change in the integrated well plate.
Specifically, the metabolite detection method can be a chromatography method, a mass spectrometry method or a chromatography-mass spectrometry combined method.
Further, the growth parameters include, but are not limited to, absorbance values in the visible range and fluorescence absorbance values.
Specifically, the method for selecting the intestinal bacteria comprises the steps of determining a target population targeted by the screening according to a candidate drug to be screened, extracting characteristic information such as relative abundance, occurrence frequency and remarkable difference degree of each bacteria according to an analysis result of the characteristics of the target population, and determining and selecting the number and the type of specific representative intestinal bacteria by taking the information as a standard so as to construct the integrated porous plate for bearing the intestinal bacteria.
Further, the growth phenotype detection judges whether the drug candidate affects the growth phenotype of the intestinal bacteria by calculating the absorbance value ratio of the plurality of intestinal bacteria processing groups added with the drug candidate and the corresponding blank control group.
Specifically, when the ratio of the absorbance values is <1.0, it is judged that the bacterial species can metabolize and utilize the compound, and when the ratio > <1.0, it is judged that the bacterial species cannot metabolize and utilize the compound.
Compared with the prior art, the invention has the beneficial effects that:
the method overcomes the defects of high cost, long period, low flux and poor repeatability of the method for evaluating the interaction of the drug and the intestinal flora based on the animal model through the high-flux screening method, has quite high flux, stability and repeatability, provides a feasible screening platform for the active drug with the action effect of the intestinal flora, avoids the shortage of funds and blindness of direction selection in the research of the prior new drug, and reduces the adverse reaction of the active drug caused by the pharmacological action generated by clinical medication.
Drawings
FIG. 1 is a flow chart of an in vitro high throughput screening platform for the effect of drug candidates on gut flora according to the present invention;
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
A. Selecting a plurality of representative intestinal strains to simulate a simplified human intestinal microbial environment according to the drug candidates and the characteristics of the intestinal flora of the target population;
b, diluting the intestinal tract strains in a microbial culture medium, and then placing the diluted intestinal tract strains in an integrated pore plate, wherein the initial concentration of the bacterial suspension of the intestinal tract strains is kept between 0.2 and 1.0;
C. respectively dripping the target drug candidates into the integrated pore plate through a microfluidic high-flux liquid distribution system, and then carrying out anaerobic culture on the intestinal strain in a continuous gas supply device;
D. culturing for 12-72 hours, and performing growth phenotype detection based on optical parameters and metabolite detection based on chromatography and mass spectrometry on the intestinal bacterial strain;
E. and according to the growth phenotype detection of the intestinal bacteria and the microbial metabolite detection of the target compound, the interaction effect of the drug candidate and the intestinal microorganisms is screened at high flux for the interaction effect of the target drug candidate and the intestinal bacteria.
Example one
In this implementationIn the case, Inosine (Inosine) is selected as a medicament to be screened, and Inosine is also called hypoxanthine nucleoside, is a biological fermentation product, can be used for the adjuvant treatment of acute and chronic hepatitis, leucopenia and thrombocytopenia caused by various reasons in medical treatment, and is also used for treating heart diseases such as pulmonary heart disease and the like and treating heart or liver toxicity reaction caused by anti-schistosome medicaments. The effect of the compound on intestinal flora is unknown, and the effect of the compound on intestinal flora of people with liver injury is expected to be researched through the screening. Determining that the in vitro intestinal flora environment to be simulated is the intestinal tract of the non-alcoholic fatty liver disease crowd, and then analyzing the high-throughput sequencing data of 324 non-alcoholic fatty liver disease human intestinal flora to determine the relative abundance of strains in the crowd sample>0.1%, frequency of occurrence in all samples>50% of 22 species of the genus Enterobacter (Alisipes, Bacteroides, Blautia, Clostridium, Coprococcus, Diarister, Dorea, Dysosmobacter, Eubacterium, Novgen _2619, Flint acter, Novgen _2710, Lachnospira, Megasphaera, Parabacteraides, Phascolatobacter, Prevotella, Roseburia, Novgen _ L23, Ruminococcus, Streptococcus, Vellonella) were selected as the species of the gut for constructing an in vitro gut bacteria screening system integrated in a multi-well plate. After selecting representative strains, respectively inoculating the strains into self-improved mGMB anaerobic culture medium for seed liquid preparation, and measuring the OD value of the culture liquid at 595nm every 12 hours after inoculation until the OD value is up to595nmNo further increase in value was considered to be complete. The growth rate of the selected gut bacteria species was scored as the length of time it took for the OD value to increase to a maximum value. The scoring criteria were as follows:
Figure BDA0002414486410000061
according to the growth rate, the seed liquid of each strain was diluted to different concentrations (as OD) with the culture medium595nmValue characterization) was used to prepare the intestinal bacterial integrated plate, with the following dilution criteria:
Figure BDA0002414486410000062
and respectively adding the intestinal strain suspensions diluted to different concentrations into a multi-hole plate, wherein each strain is added into at least 3 parallel holes, and the adding amount of each hole is 200 microliters, so as to prepare the intestinal strain integrated plate. After the screening system was constructed, inosine was added sequentially to the wells of the target multi-well plate using a microfluidic based liquid separation system to a final concentration of 20 mM. Each intestinal strain is provided with 1 blank strain control group without inosine and 2 experiment groups with inosine. Then, continuously culturing the porous plate in an anaerobic culture device with continuous gas supply for 24 hours, detecting the absorbance value in an anaerobic operation platform by a small microplate reader, and calculating the ratio (OD) of each intestinal strain treatment group added with salicin to the corresponding blank control grouptreated/ODck) Judging whether the added inosine has influence on the growth phenotype of the intestinal strain, wherein the specific standard is as follows:
Figure BDA0002414486410000063
the evaluation score of the effect of inosine on the growth of the selected 22 intestinal strains was a, and the evaluation score of 2 strains was a, that is, inosine promoted the growth of the 2 strains (Streptococcus anguinosus, Roseburia intestinalis); 1 species (Dysosmabacter welbionis) was evaluated as C, i.e., inosine inhibited the growth of the species, and the remaining 19 were evaluated as B, i.e., had no or only a weak effect on the growth of these intestinal species.
Whether the compound has influence on representative intestinal bacteria can be judged by measuring the growth phenotype, information such as a specific mode and a molecular mechanism of the influence can be further judged by detecting metabolites, and evaluation of the action mode of the most intestinal microorganisms of the target drug candidate can be finally completed by comprehensively analyzing two results. The detailed steps of metabolite detection are as follows: taking 100 microliter of fermentation liquor from each hole of the multi-hole plate subjected to growth phenotype evaluation, centrifuging, filtering, and performing liquid chromatography-tandem mass spectrometry, wherein the liquid chromatography analysis parameters are different according to different drug candidates to be detectedParameters for inosine detection were set as: c18 column, mobile phase: a is water (containing 0.5% phosphoric acid), B is methanol; the flow rate was 0.60mL/min, and the detection wavelength (. lamda.) was 248 nm. When in analysis, a standard substance of the analytical pure inosine compound is used as a positive control, and a standard curve of inosine is established at the same time; intestinal strain pure culture solution without inosine addition was used as negative control. The initial inosine content (C) was determined by measuring the concentration of inosine in a blank sample without any addition of any enteric bacteria and with 20mM inosine added to the medium aloneInitial) Subsequently, all the treatment group samples were subjected to liquid chromatography to determine the inosine concentration (C)Remainder of). Using the residual concentration C of inosine in the fermentation liquorRemainder of(mM) and initial inosine concentration CInitialThe ratio in mM measures the mode of action of intestinal species on inosine. The specific evaluation criteria are as follows:
Figure BDA0002414486410000071
in the 30 intestinal strain treatment groups, the action mode scores of 2 samples are M, and the other 20 samples are N. And then, further analyzing the chromatogram of the treatment group with the score of M, wherein the two samples have a chromatographic characteristic peak which is not contained in the initial sample group of the negative control combination, the samples with the peak-off time of the characteristic peak are collected and subjected to tandem mass spectrometry, and the mass spectrometry result is analyzed by a control database, so that the compound corresponding to the characteristic peak is hypoxanthine, the metabolism of two intestinal bacteria with inosine scored as M is indicated, and the metabolite of the two intestinal bacteria is hypoxanthine.
Performing combined analysis on the growth phenotype scoring result, the drug metabolism phenotype scoring result and the metabolite analysis result to finally give an interaction screening result of the representative intestinal strain and the target drug candidate, wherein the quantitative evaluation standard is as follows:
Figure BDA0002414486410000081
by comprehensive assessment, the action modes of inosine and 22 representative species are respectively scored as 2A + M, 1C + N and 19B + N, namely, inosine and 2 representative species (Streptococcus anguinosus, Roseburia intestinalis) have interaction, and the action mode is to promote the growth of the intestinal representative species by participating in the metabolism of the representative species to generate hypoxanthine; in addition, inosine has direct growth inhibition effect on 1 strain (Dysosmobacter welbeiis); inosine did not interact with 19 other representative species (alitistips sp., bacteriodes fragiliss, blautiassiliensis, Clostridium symbolosum, Coprococcus eutectis, Dialister sp., Dorea sp., Eubacterium elegans, Novgen _2619, Flintibacter sp., Novgen _2710, Lachnospira sp., Megasphaera sp., parabacter discosonnis, phascobacter sp., Prevotella copri, Novgen _ L23, ruminococcus biciruses, Veillonella parvula). So far, the in vitro screening platform is used for realizing high-throughput screening and comprehensive evaluation on the action effect of a drug compound inosine and typical intestinal microorganisms of nonalcoholic fatty liver patients.
Example two:
in the embodiment, salicin is selected as the drug to be screened, and the salicin has the effects of treating cold, fever and infection, relieving arthritis pain, waist and back pain and the like. The action mechanism of the compound and intestinal flora is unknown, and the effect of the compound and intestinal flora of Asian healthy people is expected to be researched through the screening. Determining that the environment of intestinal flora to be simulated in vitro is the intestinal tract of healthy Chinese people, and then analyzing the high-throughput sequencing data of 516 intestinal flora of healthy people to determine the relative abundance of strains in a population sample>0.1%, frequency of occurrence in all samples>50% of 30 enterobacteria Alistepes, Bacteroides, Bifidobacterium, Blautia, Butyricicoccus, Clostridium, Collinsela, Coprococcus, Dyssomobacter, Eubacterium, Faecalibacillus, Intestibacter, Lactobacilli, Novgen _2710, Odoribacter, Oscilobacter, Parabacter, Phascolatobacter, Prevotella, Romboutsia, Roseburia, Ruminococcus, Streptomyces, Absysivirga, Anaetreruccus, Catenibacillus, Cellulosilyticum, Christensella, Mitsukelella were selected from each genus as an enterobacteria strain for in vitro selection of multi-well plates. At the time of selectionAfter representative strains, the strains are respectively inoculated into self-improved mGMB anaerobic culture medium for seed solution preparation, and OD values of the culture solution at 595nm are measured every 12 hours after inoculation until OD is reached595nmNo further increase in value was considered to be complete. The growth rate of the selected gut bacteria species was scored as the length of time it took for the OD value to increase to a maximum value. The scoring criteria were as follows:
Figure BDA0002414486410000101
according to the growth rate, the seed liquid of each strain was diluted to different concentrations (as OD) with the culture medium595nmValue characterization) was used to prepare the intestinal bacterial integrated plate, with the following dilution criteria:
Figure BDA0002414486410000102
and respectively adding the intestinal strain suspensions diluted to different concentrations into a multi-hole plate, wherein each strain is added into at least 3 parallel holes, and the adding amount of each hole is 200 microliters, so as to prepare the intestinal strain integrated plate. After the screening system was constructed, salicin was sequentially added to the wells of the target multi-well plate using a microfluidic based liquid separation system to a final concentration of 50 mM. Each intestinal bacterial strain is provided with 1 blank bacterial strain control group without salicin and 2 experimental groups with salicin. Then, the porous plate is continuously cultured for 48 hours in an anaerobic culture device with continuous gas supply, then the absorbance value is detected in an anaerobic operation platform by a small microplate reader, and the ratio (OD) of each intestinal strain treatment group added with salicin to the corresponding blank control group is calculatedtreated/ODck) Judging whether the added salicin has influence on the growth phenotype of the intestinal bacteria, and the specific standard is as follows:
Figure BDA0002414486410000111
the judgment score of the effect of the salicin on the growth of the selected 30 intestinal strains is that the evaluation score of the 13 strains is A, namely the growth of the 13 strains is promoted; the remaining 17 were judged as B, i.e. had no or only a weak effect on the growth of these intestinal species; there were no intestinal species scored as C.
Whether the compound has influence on representative intestinal bacteria can be judged by measuring the growth phenotype, information such as a specific mode and a molecular mechanism of the influence can be further judged by detecting metabolites, and evaluation of the action mode of the most intestinal microorganisms of the target drug candidate can be finally completed by comprehensively analyzing two results. The detailed steps of metabolite detection are as follows: from each well of the multi-well plate after growth phenotype evaluation, 100. mu.l of fermentation broth was subjected to a liquid chromatography-tandem mass spectrometry analysis after centrifugation, with the liquid chromatography analysis parameters set to C18 column, in methanol: water (10: 90) was used as the mobile phase, the flow rate was 1.0mL/min, and the detection wavelength was 269 nm. During analysis, a salicin compound standard substance is used as a positive control, a salicin standard curve is established, and a pure intestinal strain culture solution without salicin is used as a negative control. Firstly, measuring the salicin concentration corresponding to a blank sample which is not added with any intestinal canal strain and is only added with 50mM salicin in a culture medium as an initial salicin content value, and then sequentially carrying out liquid chromatography on all treatment group samples to measure the salicin concentration. The action mode of intestinal bacteria to salicin is measured by the ratio of the residual salicin concentration (mM) to the initial salicin concentration (mM) in the fermentation broth. The specific evaluation criteria are as follows:
Figure BDA0002414486410000121
the treatment groups of 30 intestinal species at this time, wherein 13 samples had an action pattern score of M and the remaining 17 were N. And then, further analyzing the chromatograms of the treatment groups with the scores of M, and if a chromatographic characteristic peak which is not existed in the initial sample group of the negative control combination is found, carrying out subsequent tandem mass spectrometry to determine which product the target candidate drug is metabolized by the strains. (in this example, no subsequent tandem mass spectrometry was performed since no new characteristic peak was generated in the chromatogram for all samples scored by M).
And (3) carrying out combined analysis on the growth phenotype scoring result and the drug metabolism phenotype scoring result to finally give a screening result of the interaction between the representative intestinal strain and the target drug candidate, wherein the quantitative evaluation standard is as follows:
Figure BDA0002414486410000122
by comprehensive assessment, the mode of action of salicin and 30 representative species is scored as 13 a + M and 17B + N, respectively, i.e., salicin interacts with 13 representative species (Abyssivirga sp., anaerobactuncus sp., bacteroides fragilis, Blautia maliensis, catebiaceae sp., celluliticum sp., christensella sp., Clostridium symbolosum, coprococus sp., Mitsuokella sp., paramylobacter disco, Roseburia sp., strepcoccus sp.) by participating in the metabolism of these representative species to promote the growth of these intestinal representative species; in addition, salicin did not interact with 17 other representative species (alitistips sp., Bifidobacterium bifidum, butyricucusfaecionis, collinesila _ intestinalis, dysmicrobacterium welblinis, eubacterium elegins, faecibium sp., intestinobacterium sp., Lactobacillus sp., Novgen _2710, oribacter sp., Oscillibacter sp., phascobacter sp., Prevotella copri, rombouutosia sp., ruminococcus bicirus). So far, the in vitro screening platform is used for realizing high-throughput screening and comprehensive evaluation on the action effect of a target drug candidate salicin and typical intestinal microorganisms of Asian healthy people.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims (7)

1. An in vitro high-throughput screening platform for the effect of drug candidates on intestinal flora, comprising the steps of:
A. selecting a plurality of representative intestinal strains to simulate a simplified human intestinal microbial environment according to the drug candidates and the characteristics of the intestinal flora of the target population;
b, diluting the intestinal tract strains in a microbial culture medium, and then placing the diluted intestinal tract strains in an integrated pore plate, wherein the initial concentration of the bacterial suspension of the intestinal tract strains is kept between 0.2 and 1.0;
C. respectively dripping the target drug candidates into the integrated pore plate through a microfluidic high-flux liquid distribution system, and then carrying out anaerobic culture on the intestinal strain in a continuous gas supply device;
D. culturing for 12-72 hours and performing phenotype detection and metabolite detection based on growth parameters on the intestinal bacteria;
E. and according to the growth phenotype detection of the intestinal bacteria and the microbial metabolite detection of the target compound, the high-throughput screening of the interaction effect of the drug candidate and the intestinal microorganisms is realized.
2. The platform of claim 1, wherein said metabolite testing reflects the effect of different drug candidates on the activity of said gut bacteria metabolite testing by altering the biomass of said gut bacteria species in an integrated well plate.
3. The platform for in vitro high-throughput screening of effect of drug candidates on gut flora according to claim 1, wherein the metabolite detection method is chromatography, mass spectrometry or a combination of chromatography and mass spectrometry.
4. The platform of claim 1, wherein the growth parameters include but are not limited to absorbance values in the visible range and fluorescence absorbance values.
5. The in vitro high-throughput screening platform for the effect of drug candidates on intestinal flora according to claim 1, wherein the selection method of intestinal bacteria species comprises the steps of determining a target population targeted by the current screening according to the drug candidates to be screened, extracting characteristic information such as relative abundance, occurrence frequency and degree of significant difference of each bacteria species according to the analysis result of the characteristics of the target population, and determining and selecting the number and the type of specific representative intestinal bacteria species according to the information as a standard to construct the integrated multi-well plate for bearing the intestinal bacteria species.
6. The in vitro high-throughput screening platform for effect of drug candidate on intestinal flora according to claim 1, wherein said growth phenotype detection determines whether said drug candidate affects the growth phenotype of intestinal flora by calculating absorbance ratio of multiple said intestinal flora treatment groups added with drug candidate and its corresponding blank control group.
7. The platform of claim 6, wherein the absorbance value ratio is <1.0, the bacterial species is determined to be able to metabolize and utilize the compound, and the ratio > is 1.0, the bacterial species is determined to be not able to metabolize and utilize the compound.
CN202010186816.3A 2020-03-17 2020-03-17 In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora Pending CN111307982A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010186816.3A CN111307982A (en) 2020-03-17 2020-03-17 In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010186816.3A CN111307982A (en) 2020-03-17 2020-03-17 In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora

Publications (1)

Publication Number Publication Date
CN111307982A true CN111307982A (en) 2020-06-19

Family

ID=71151274

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010186816.3A Pending CN111307982A (en) 2020-03-17 2020-03-17 In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora

Country Status (1)

Country Link
CN (1) CN111307982A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115232855A (en) * 2022-07-08 2022-10-25 华南农业大学 Method for screening drugs influencing xanthine oxidase activity by targeting intestinal flora

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150376716A1 (en) * 2013-02-12 2015-12-31 Japanese Foundation For Cancer Research Method for examination of carcinogenic risk
CN109439722A (en) * 2018-10-24 2019-03-08 浙江工商大学 Measuring method of the lactic acid bacteria based on gut simulation model to enteron aisle prebiotic effect
CN109706111A (en) * 2019-02-21 2019-05-03 中山大学 The quick screening model and its construction method of P. aeruginosa bacteria quorum sensing system inhibitor
CN110643686A (en) * 2019-10-24 2020-01-03 上海交通大学医学院附属仁济医院 Method for screening drugs with potential microorganism regulation ability in vivo

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150376716A1 (en) * 2013-02-12 2015-12-31 Japanese Foundation For Cancer Research Method for examination of carcinogenic risk
CN109439722A (en) * 2018-10-24 2019-03-08 浙江工商大学 Measuring method of the lactic acid bacteria based on gut simulation model to enteron aisle prebiotic effect
CN109706111A (en) * 2019-02-21 2019-05-03 中山大学 The quick screening model and its construction method of P. aeruginosa bacteria quorum sensing system inhibitor
CN110643686A (en) * 2019-10-24 2020-01-03 上海交通大学医学院附属仁济医院 Method for screening drugs with potential microorganism regulation ability in vivo

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
LISA MAIER 等: "Extensive impact of non-antibiotic drugs on human gut bacteria", 《NATURE》 *
MICHAEL ZIMMERMANN 等: "Mapping human microbiome drug metabolism by gut bacteria and their genes", 《NATURE》 *
史权 等: "对大鼠肠道菌群有调整作用的中药筛选", 《黑龙江医药科学》 *
张怡红 等: "离体培养人肠道菌群对黄山药总皂苷的代谢研究", 《中国现代药物应用》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115232855A (en) * 2022-07-08 2022-10-25 华南农业大学 Method for screening drugs influencing xanthine oxidase activity by targeting intestinal flora

Similar Documents

Publication Publication Date Title
Sellick et al. Effective quenching processes for physiologically valid metabolite profiling of suspension cultured mammalian cells
Kristoffersen et al. Headspace gas chromatographic determination of ethanol: the use of factorial design to study effects of blood storage and headspace conditions on ethanol stability and acetaldehyde formation in whole blood and plasma
Andreoli et al. Reference ranges of urinary biomarkers of oxidized guanine in (2′-deoxy) ribonucleotides and nucleic acids
US20160116461A1 (en) Biomarkers Related To Metabolic Age and Methods Using The Same
Møller et al. Harmonising measurements of 8-oxo-7, 8-dihydro-2′-deoxyguanosine in cellular DNA and urine
CN110366596A (en) For being analyzed the Whole microbial strain in complex heterogeneous group, determining its functional relationship and interaction and being identified based on this and the method, apparatus and system of synthesis of biologically active modifying agent
JP2016513259A (en) Method for identifying bacterial species in a biological sample by gas chromatography mass spectrometry (GC / MS)
Manoni et al. Field evaluation of a second-generation cytometer UF-100 in diagnosis of acute urinary tract infections in adult patients
JPH10513048A (en) Screening for modulators of biomolecules
AU2018235992B2 (en) Device, method, and system for identifying organisms and determining their sensitivity to toxic substances using the changes in the concentrations of metabolites present in growth medium
Pietersen et al. Tween 80 induces a carbon flux rerouting in Mycobacterium tuberculosis
CN112505179B (en) Method for measuring isotope dilution ultra-performance liquid chromatography-mass spectrometry combination
CN111307982A (en) In-vitro high-throughput screening platform for effect of drug candidate on intestinal flora
CN105004825A (en) GC/MS metabonomics analysis method based on osteoblast cell tissue
Levin et al. Discovering radical-dependent enzymes in the human gut microbiota
Velivasi et al. Modeling postmortem ethanol production by C. albicans: Experimental study and multivariate evaluation
Gao et al. Flow injection spectrophotometric determination of sulfated bile acids in urine with immobilized enzyme reactors using water soluble tetrazolium blue-5
CN117210527A (en) Tumor organoid drug-resistant clone-based clinical drug-resistant post-treatment guidance system
Lagat et al. Development of an ELISA-based method for testing aflatoxigenicity and aflatoxigenic variability among Aspergillus species in culture
CN109187814A (en) A kind of method and its detection kit for identifying kidney transplant prognosis biomarker
Lee et al. Experimental design in metabolomics
CN111370116A (en) Intestinal microbial marker for predicting curative effect of bipolar affective disorder and screening application thereof
Adekanye et al. Abstract 2073 KNOCKDOWN OF CES1 IN THP-1 MACROPHAGES PROMOTES A PROINFLAMMATORY PHENOTYPE
WO2021152936A1 (en) Method and system for analyzing metabolites
Miralles et al. In vitro culture and biochemical characterization of six trypanosome isolates from Peru and Brazil

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
RJ01 Rejection of invention patent application after publication

Application publication date: 20200619

RJ01 Rejection of invention patent application after publication