CN110507823A - Application of the itaconic acid related substances in diagnosis, prevention and treatment disease of viral infection - Google Patents

Application of the itaconic acid related substances in diagnosis, prevention and treatment disease of viral infection Download PDF

Info

Publication number
CN110507823A
CN110507823A CN201910769226.0A CN201910769226A CN110507823A CN 110507823 A CN110507823 A CN 110507823A CN 201910769226 A CN201910769226 A CN 201910769226A CN 110507823 A CN110507823 A CN 110507823A
Authority
CN
China
Prior art keywords
itaconic acid
virus
disease
leu
infection
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
Application number
CN201910769226.0A
Other languages
Chinese (zh)
Other versions
CN110507823B (en
Inventor
刘洋
曹雪涛
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 Basic Medical Sciences of CAMS
Original Assignee
Institute of Basic Medical Sciences of CAMS
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 Basic Medical Sciences of CAMS filed Critical Institute of Basic Medical Sciences of CAMS
Priority to CN201910769226.0A priority Critical patent/CN110507823B/en
Publication of CN110507823A publication Critical patent/CN110507823A/en
Application granted granted Critical
Publication of CN110507823B publication Critical patent/CN110507823B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/705—Specific hybridization probes for herpetoviridae, e.g. herpes simplex, varicella zoster
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/706—Specific hybridization probes for hepatitis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/136—Screening for pharmacological compounds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Virology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Communicable Diseases (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Epidemiology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

This disclosure relates to application of itaconic acid (Itaconic acid) related substances in diagnosis, prevention and treatment disease of viral infection.Specifically, this disclosure relates to itaconic acid mortifier is preparing the purposes in the product for preventing and/or treating disease of viral infection and/or disease relevant to virus infection and/or symptom, and its corresponding pharmaceutical composition or medicine box.Present disclosure also relates to detect application of the reagent of levels of substance in itaconic acid and/or its metabolic pathway in the kit for preparing judging viral infection disease or its gradient of infection.

Description

Application of the itaconic acid related substances in diagnosis, prevention and treatment disease of viral infection
Technical field
The disclosure belongs to biotechnology and medical domain.Specifically, this disclosure relates to itaconic acid mortifier is cured the disease anti- Levels of substance is in detection virus infection and its journey in application and itaconic acid and/or its metabolic pathway in malicious infectious diseases Application in degree.
Background technique
Virus infection can cause a variety of diseases and threaten human health.Research virus-host's interaction mechanism facilitates depth Enter to understand virus infection and its regulatory molecule mechanism, and new medicine target is provided for prevention and treatment disease of viral infection Point.
Innate immunity receptor can activate the kinases in downstream signaling pathway by the virus of identification infection host cell Thus TBK1 and transcription factor IRF3 etc. induce body to generate interferon (Interferon, IFN) and activate antiviral effect base The expression of cause with resist and remove virus (Schneider WM etc., Annual review of immunology, 2014,32: 513-545)。
In addition to this, cell metabolism also assists in the biological processes such as regulation virus and host's interaction.When identifying or sense After the virus of invasion, host cell can remold the metabolism state of itself energetically to generate substance necessary to viral survival Promote virus infection, or realizes the removing to virus by suppressing virus replication.This process may rely on or not depend on In activation antiviral natural immune response (Phan AT etc., Immunity, 2017,46 (5): 714-729；Olive AJ etc., Nat Rev Microbiol,2016,14(4):221-234).For example, the lncRNA-ACOD1 of the induced generation of virus infection can lead to It crosses and directly combines and promote with metabolic enzyme aspartate transaminase 2 (Glutamic-oxaloacetic transaminase 2, GOT2) Aspartic acid (Aspartate) and α-ketoglutaric acid (α-ketoglutarate) needed for its enzymatic activity carrys out enhanced virus duplication It generates, to promote virus infection (Wang P etc., Science, 2017,358 (6366): 1051-1055).
On the other hand, virus can also complete its life cycle by adjusting the metabolic pathway of host cell, help disease The monitoring of poison escape immune system, and an advantageous cellular environment is created to guarantee that viral persistence constantly infects.For example, gland Virus infection can change the metabolic pathway of cell, and the metabolic pathway of induction Warburg sample is converted into aerobic glycolysis (Aerobic glycolysis), nucleotide needed for synthesizing some virus replications with this and protein (Thai M etc., Cell metabolism,2014,19(4):694-701)。
How host cell passes through adjusting metabolism remodeling process, builds especially by the mode independent of immune response The vertical tolerance to virus infection still has many unknown with resistance state.In addition, participate in regulation virus infection and duplication it is new Metabolin be also required to more study to be identified.
Small molecule metabolites itaconic acid (Itaconic acid), also referred to as methene succinic acid, methylene-succinic acid are a kind of Active binary organic acid containing unsaturated double-bond, molecular formula C5H6O4, there is molecular structural formula as follows:
In vivo in metabolic process, itaconic acid is transformed by aconitate (Cis-aconitate), and the latter is tricarboxylic acids Recycle one of the important intermediate of (TCA circulation).Aconitate decarboxylase is by 1 (Immune- of immune response gene Responsive gene 1, IRG1) coding a kind of metabolic enzyme, using aconitate as substrate directly be catalyzed generation itaconic acid (Michelucci A etc., Proc Natl Acad Sci U S A.2013,10 (19): 7820-7825).In lipopolysaccharides In the case of (Llipopolysaccharide, LPS) stimulation, itaconic acid is that most significant generation is raised in people and mouse macrophage Thank to object.Under LPS stimulation, itaconic acid can be by being alkylated KEAP1 come activating transcription factor Nrf2 to play anti-inflammatory effect (Mills EL etc., Nature, 2018,556 (7699): 113-117)；Itaconic acid can also be by inhibiting succinate dehydrogenase (succinate dehydrogenase, SDH) simultaneously inducing cell metabolism is remolded to adjust mitochondrial respiratory, to play inhibition Effect (LampropoulouV etc., the Cell Metab.2016 of inflammation；24(1):158-66).
Currently, there is no the effect in relation to itaconic acid in virus infection and regulatory mechanism both at home and abroad, and its in virus Research report in terms of diseases related middle treatment use.And there is an urgent need in the art to search out disease of viral infection treatment Potential important target molecule is being faced by its clear being associated with disease of viral infection with promoting and reinforcing the target molecules Application in bed treatment.
Summary of the invention
One of main purpose of the disclosure is to disclose the correlation between itaconic acid and virus infection, and provides using clothing Health acid mortifier come treat the application of disease of viral infection and/or disease relevant to virus infection and/or symptom, method and Product, and by detection itaconic acid and/or its metabolic pathway in substance level come judging viral infection disease and/or its Application, method and the product of gradient of infection.
In in one aspect of the present disclosure, itaconic acid mortifier is provided in preparation in prevention and/or treatment object Purposes in the product of disease of viral infection and/or disease relevant to virus infection and/or symptom.
In in one aspect of the present disclosure, itaconic acid mortifier is provided, is used for virus in prevention and/or treatment object Infectious diseases and/or disease relevant to virus infection and/or symptom.
In in one aspect of the present disclosure, provide in treatment object disease of viral infection and/or with virus infection phase The method of the disease and/or symptom of pass, the method includes giving subject in need a effective amount of itaconic acid mortifier.
In some embodiments, the itaconic acid mortifier is selected from the mortifier for following substance: itaconic acid, clothing health Enzyme needed for acid metabolic precursor (such as aconitate), metabolism generate itaconic acid (such as aconitate decarboxylase IRG1, α -one penta Two acidohydrogenase OGDH).
In some embodiments, the itaconic acid mortifier is selected from: the upstream generation generated for itaconic acid or its precursor Thank the antibody (such as antibody for IRG1 or OGDH) of enzyme, siRNA (such as in SEQ ID NO:3-8 it is any shown in be directed to The siRNA of OGDH), miRNA, antisense oligonucleotides, CRISPR/Cas9 gene editing product, antagonist, blocking agent, inhibit clothing Health acid or its preceding body function and/or active compound lower enzyme (such as OGDH or IRG1) needed for metabolism generates itaconic acid The promoter element and/or expression vector of gene or mRNA or protein expression level.
In some embodiments, itaconic acid metabolic precursor thereof is selected from: the substance of itaconic acid can be generated through processing in vivo, it is such as suitable Aconitic acid.
In some embodiments, infect for DNA virus and/or picornavirus infection, for example, by selected from the group below a kind of or Infection caused by a variety of viruses: herpes simplex virus, vesicular stomatitis virus, influenza virus, encephalomyocarditis virus, celestial platform disease Poison, hepatitis type B virus, adenovirus, poxvirus, small DNA virus, adeno-associated virus.
In some embodiments, disease relevant to virus infection and/or symptom are one of to be selected from the group or more Kind: pathology damage caused by virus infection；The cells following viral infection factor (such as interferon) generates insufficient or excessive；Endotoxin induction Shock is dead；The inflammatory damage of organ；Multiple organ failure, such as the organ are selected from: liver, spleen, brain, kidney, the heart Dirty, lung, stomach, intestines；Chronic inflammation disease caused by virus infection (such as autoimmune disease such as inflammatory bowel disease, class wind Wet arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, Chronic gastrointestinal diseases).
In some embodiments, the object is mammal, such as people, pet, domestic animal, such as with the innate immunity Defect/impaired and/or interferon anti-reflecting virus treatment of infection is invalid or ineffective or is also easy to produce the object of side effect, such as with certainly Body immunity disease or the object for being susceptible to suffer from autoimmune disease, as IFN insufficiency or too drastic object (such as suffer from inflammatory bowel Disease, rheumatoid arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, Chronic gastrointestinal diseases object).
In some embodiments, the product is pharmaceutical composition or medicine box, such as its form is suitable for by being selected from down The pharmaceutical composition or medicine box that group mode is given: oral, injection (such as direct naked DNA or protein injection method, liposome DNA or protein injection method), gold coating particle bombardment, breeding unsoundness bacterium carry Plasmid DNA method, replication defective adenopathy Poison carries protein coded by target DNA method or target gene, electroporation, nasal-cavity administration, pulmonary administration, oral administration, transdermal Administration in administration, tumor.
In some embodiments, product also include for prevent and/or treat disease of viral infection and/or with virus One of other substances of relevant disease and/or symptom are infected, such as are selected from the group or is a variety of: clinical common antibiotics, Including beta-lactam (penicillins and cephalosporins), aminoglycosides, Tetracyclines, chloromycetin, macrolide, Antifungal antibiotic, treating tuberculosis class antibiotic；(tricyclic amines, pyrophosphoric acid class, protease inhibit the common antiviral drugs of clinic Medicine, nucleoside medicine and interferon, antisense oligonu-cleotides etc.)；The common immunosuppressor of clinic (including glucocorticoid, ring Phosphamide, chloroquine, Ciclosporin A, tripterygium wilfordii, Chinese materia medica preparation, anti-TNF monoclonal antibody).
In the another aspect of this paper, a kind of pharmaceutical composition or medicine box are provided, it includes:
(A) the itaconic acid mortifier of prevention and/or therapeutically effective amount；
(B) acceptable carrier or excipient pharmaceutically or in immunology；
(C) optionally, one or more preventions and/or treatment disease of viral infection or disease relevant to virus infection Other active materials of disease and/or symptom.
In some embodiments, the mortifier and other substances are as described in disclosure context.
In some embodiments, itaconic acid mortifier account for pharmaceutical composition or medicine box total weight 0.001~ 99.9wt%.
In some embodiments, itaconic acid mortifier accounts for 1~95wt% in pharmaceutical composition or medicine box total weight, and 5 ~90wt%, 10~80wt%, surplus are pharmaceutically acceptable carrier, other active materials or other additions that may be present The substances such as agent.
In some embodiments, it prior to, concurrently with, or after the pharmaceutical composition or medicine box for giving the disclosure, gives and adjusts Control other active materials of viral infection resisting.Other active materials have prevention or treatment disease relevant to virus infection The activity of chronic inflammation disease caused by damaging, infecting caused by disease, infection and/or its symptom.The virus infection is DNA Virus and/or picornavirus infection, such as the infection caused by one or more viruses selected from the group below: herpes simplex virus, water Bubble property Stomatovirus, influenza virus, encephalomyocarditis virus, sendai virus, hepatitis type B virus, adenovirus, poxvirus, small DNA Virus, adeno-associated virus.
In in one aspect of the present disclosure, levels of substance in detection itaconic acid body and/or its relevant metabolic pathway is provided Application of the reagent in the kit for preparing judging viral infection disease or its gradient of infection.
In in one aspect of the present disclosure, a kind of judging viral infection disease or the method for its gradient of infection are provided, The method includes levels of substance in detection itaconic acid and/or its relevant metabolic pathway, and according to the variation of the level and/or height It is low come judging viral infection disease or its gradient of infection, wherein levels of substance mentions in itaconic acid and/or its relevant metabolic pathway Height shows the presence of disease of viral infection, and the height improved the standard is corresponding with gradient of infection height.
In in one aspect of the present disclosure, levels of substance in detection itaconic acid and/or its relevant metabolic pathway is provided Reagent is used for judging viral infection disease or its gradient of infection.
In some embodiments, substance is selected from the itaconic acid and/or its relevant metabolic pathway: itaconic acid, clothing health Acid metabolic precursor (such as itaconate, aconitate), itaconic acid metabolite, itaconic acid generate needed for enzyme (such as IRG1 or OGDH)。
In in one aspect of the present disclosure, a kind of kit is provided comprising: detection is obtained from the sample underpants health of object The reagent of levels of substance in acid and/or its relevant metabolic pathway.
In in one aspect of the present disclosure, a kind of screen by inhibiting itaconic acid come the drug of viral infection resisting is provided Method comprising:
(A) infected cell, tissue or animal are handled with candidate substances；
(B) detect the cell, itaconic acid in tissue or animal, its metabolic precursor thereof (such as itaconate, aconitate), Itaconic acid metabolite, itaconic acid generate the level or activity of required enzyme (such as IRG1 or OGDH)；And
(C) if level or activity detected is lower than the respective horizontal or activity before candidate substances processing or lower than normal The infection of respective horizontal or activity and the cell, tissue or animal in control is controlled or is suppressed, then shows described Candidate substances may be as by inhibiting itaconic acid come the drug of viral infection resisting.
In some embodiments, OGDH the or IRG1 level is its DNA, mRNA or protein level.
In some embodiments, the method still further comprise detection the candidate substances processing in or later, The variation of IFN level in cell, tissue or the animal being infected.If IFN level is without significant changes or significant decrease, and disease Poison infection is controlled or is inhibited, then shows that the safety of the candidate substances is higher, in particular for innate immunity imbalance or IFN Safety for insufficiency or autoimmune disease object is higher.
Those skilled in the art can carry out any combination without departing from this public affairs to technical solution above-mentioned and technical characteristic The inventive concept opened and protection scope.The other aspects of the disclosure are due to this disclosure, to those skilled in the art For be obvious.
Detailed description of the invention
The disclosure is described further with reference to the accompanying drawing, wherein these show only for illustrating the reality of the disclosure Scheme is applied, rather than in order to limit to the scope of the present disclosure.In figure, " * " indicates P < 0.05, and " * * " indicates P < 0.01, and " * * * " is indicated P < 0.001, " * * * * " indicate P < 0.0001, and there was no significant difference for " NS " expression, and (double non-paired student t of tail are examined, Two- tailed unpaired Student’s t-test)。
Fig. 1: the mouse primary peritoneal macrophage underpants health acid content of virus infection significantly increases.
Fig. 2: itaconic acid can promote the virus replication in mouse primary peritoneal macrophage in a dose-dependent manner:
A: itaconic acid class compound dimethyl itaconate (Dimethyl itaconate, DMI) can be in a dose-dependent manner Promote the virus replication in mouse primary peritoneal macrophage；
B: itaconic acid class compound 4- octyl itaconic acid (4-Octyl itaconate, OI) can promote in a dose-dependent manner Virus replication into mouse primary peritoneal macrophage.
Fig. 3: itaconic acid has the function of promoting virus replication in Mice Body.
Fig. 4: itaconic acid promotes Mice Body inner virus to infect by the non-dependent mode of interferon
A: itaconic acid promotes the IFN-β protein expression that virus infection is induced in mice serum；
B: itaconic acid promotes the IFN-α protein expression that virus infection is induced in mice serum.
Fig. 5: it strikes low metabolic enzyme OGDH expression and can lower cell underpants health acid content and carry out suppressing virus replication:
A: the jamming effectiveness of the siRNA (siRNA) of targeting mouse OGDH；
B: metabolin itaconic acid and suitable crow in mouse primary peritoneal macrophage can be lowered by striking low metabolic enzyme OGDH expression The content of head acid；
C: low OGDH expression is struck in mouse primary peritoneal macrophage can inhibit virus replication (left figure), which does not depend on To interferon response (right figure).
Specific embodiment
The present inventor passes through long-term in-depth study discovery: metabolin itaconic acid in mouse cell and Mice Body in The effect for remarkably promoting virus infection and duplication is all had, this implies the strategy that targeted inhibition itaconic acid is generated or played a role The potential using value of effective therapy target as control virus infection.
On the one hand, pass through (such as itaconic acid metabolic precursor thereof (such as clothing in detection itaconic acid and/or its relevant metabolic pathway Health acid esters, aconitate), itaconic acid metabolite, itaconic acid generate needed for enzyme) level or activity method with it is related try Agent box, to judge whether body is infected and gradient of infection.This is for the diagnosis of viral infection related disease and pre- It is anti-to have significant application value.
On the other hand, using itaconic acid mortifier, such as the suppression of upstream metabolic enzyme IRG1 and/or OGDH that itaconic acid generates Preparation, siRNA and CRISPR/Cas9 gene editing product, clothing for itaconic acid upstream metabolic enzyme IRG1 and/or OGDH Health acid metabolic pathway inhibitor and/or specific antibody, come prepare for prevent and/or treat infectious diseases and/or with sense It contaminates the product of relevant disease and/or symptom, thus itaconic acid is prevented to generate or function, suppressing virus replication protects machine Body is from virus infection.Its corresponding pharmaceutical composition or medicine box, the disclosure can be used for preventing and treating viral infection correlation Disease has extensive potential applicability in clinical practice.
By being shown the results show that using itaconic acid as drug target for the following viral infection clinically Disease treatment application potential has very big directive significance especially for the patient that innate immune function is damaged.The strategy Another advantage is that, the excess generation of interferon can be avoided using the treatment of itaconic acid blocked method, and avoid day Inflammatory autoimmune disease caused by right immune activation.Therefore, the method for targeted inhibition itaconic acid has stronger general Adaptive and safety.
Accordingly, this disclosure relates to which itaconic acid and the correlation and its inhibitor of virus infection pass through non-natural immune dependent Property effect reach the application of antiviral effect, and by detection itaconic acid content come judging viral infection and gradient of infection Methods and applications.Specifically, this disclosure relates to itaconic acid mortifier is in preparation for preventing and/or treating infectious diseases And/or the purposes in the product of disease relevant to infection and/or symptom, corresponding treatment method, product.The disclosure also relates to And the substance of detection itaconic acid level is preparing the purposes in the product for diagnosing infectious diseases and/or its gradient of infection, Its corresponding diagnostic method and product.The disclosure can be used for diagnosing, prevent and/or treat viral infection related disease, have Broad application prospect.
All numberical ranges provided herein be intended to clearly include fall in all numerical value between endpoints of ranges and it Between numberical range.The feature that can be mentioned to the feature or embodiment that the disclosure is mentioned is combined.This specification is taken off All features shown can be used in combination with any composition form, and each feature disclosed in specification any can provide phase The alternative characteristics of same, impartial or similar purpose replace.Therefore except there is special instruction, revealed feature is only impartial or similar The general example of feature.
As used herein, " containing ", " having " or " comprising " include "comprising", " mainly by ... constitute ", " substantially By ... constitute " and " by ... constitute "；" mainly by ... constitute ", " substantially by ... constitute " and " by ... constitute " Belong to the subordinate concept of " containing ", " having " or " comprising ".
Itaconic acid related substances
As used herein, term " itaconic acid related substances " refers to when being related to diagnostic context for detecting itaconic acid And/or in its relevant metabolic pathway substance reagent, the substance in these metabolic pathways includes but is not limited to: before itaconic acid metabolism Enzyme needed for body (such as itaconate, aconitate), itaconic acid metabolite, itaconic acid generation (such as such as aconitate Decarboxylase IRG1, ketoglurate dehydrogenase OGDH).
As used herein, term " itaconic acid related substances ", can be with " itaconic acid suppression when being related to preventing and treating context Object processed " is used interchangeably, be refer to directly to inhibit itaconic acid and/or by inhibit in itaconic acid relevant metabolic pathway substance to Inhibit the substance of itaconic acid indirectly.In some embodiments, substance can be for example but unlimited in itaconic acid relevant metabolic pathway In following substance: enzyme needed for itaconic acid metabolic precursor thereof (such as aconitate), itaconic acid metabolite, metabolism generate itaconic acid (such as aconitate decarboxylase IRG1, ketoglurate dehydrogenase OGDH) etc..
In some embodiments, the itaconic acid mortifier is selected from: inhibit itaconic acid substance (such as compound, such as Small molecule compound), the antibody of the upstream metabolic enzyme generated for itaconic acid or its precursor (such as the anti-of IRG1 or OGDH Body), siRNA (such as in SEQ ID NO:3-8 it is any shown in be directed to OGDH siRNA), miRNA, antisense oligonucleotides, CRISPR/Cas9 gene editing product, blocking agent, inhibits itaconic acid or its preceding body function and/or active chemical combination at antagonist Object lowers the gene of enzyme (such as OGDH or IRG1) or the promoter of mRNA or protein expression level needed for metabolism generates itaconic acid Element and/or expression vector.
By taking the ketoglurate dehydrogenase (OGDH) in itaconic acid metabolic pathway as an example, for during tricarboxylic acid cycle First rate-limiting enzyme.Through the verification experimental verification in the disclosure, the level and activity of OGDH can influence itaconic acid and its precursor along the rhizome of Chinese monkshood The level and activity of acid, by strike the expression of the metabolic enzyme OGDH in low itaconic acid metabolic pathway can lower in cell itaconic acid with Aconitate content is to play the role of suppressing virus replication, control virus infection.
OGDH polypeptide is well known in the art, such as it is in mouse [Mus musculus (house mouse)] Corresponding to NCBI Gene ID:18293, amino acid sequence can be as shown in SEQ ID NO:12；It is in people [Homo sapiens (human)] correspond to NCBI Gene ID:4967 in, amino acid sequence can be as shown in SEQ ID NO:14.For generating The nucleic acid molecules of OGDH polypeptide refer to the nucleic acid molecules that OGDH albumen or polypeptide can be obtained by it.The nucleic acid molecules can for Such as: OGDH genomic dna sequence (see, for example, https: //www.ncbi.nlm.nih.gov/nuccore/NC_ 000077.6 from=6291597&to=6359094&report=genbank (mouse, Mus musculus) or Https: //www.ncbi.nlm.nih.gov/nuccore/NG_023260.1 from=5001&to=107549&rep Ort=genbank (homo sapiens, Homo sapiens)), its transcript profile mRNA sequence it is (such as (small as shown in SEQ ID NO:11 Mouse, Mus musculus) or as shown in SEQ ID NO:13 (homo sapiens Homo sapiens)) or its precursor or variant or homologous sequence Column.
Those of ordinary skill in the art can provide its corresponding antibody, siRNA (such as SEQ ID NO:3-8 for OGDH In it is any shown in), miRNA, antisense oligonucleotides, CRISPR/Cas9 gene editing product etc., by blocking or inhibiting the generation Thank to expression or the activity of enzyme to reduce the generation of itaconic acid or inhibit itaconic acid activity, to generate prevention and/or treatment virus The effect of infectious diseases.
Those of ordinary skill in the art can be can promote virus replication and be inhibited clothing based on the principle of the disclosure, i.e. itaconic acid Health acid can inhibit virus replication and infection, can get or design the various mortifiers for being directed to itaconic acid.
The mortifier of the disclosure can effectively block by non-natural immune dependent sexual approach or inhibit itaconic acid multiple to virus System, proliferation and infection facilitation, caused by avoiding because of the innate immunity dependence treatment interferon excess generation and because The caused inflammatory autoimmune disease of innate immunity activation.Therefore, the mortifier of the disclosure and method have stronger Universality and safety.
The antiviral mode of the innate immunity (or interferon) dependent/non-dependent
As described in the background section, in virus infection, innate immunity receptor can be thin by identification infection host The virus of born of the same parents activates the kinases TBK1 in downstream signaling pathway and transcription factor IRF3 etc., thus body is promoted to generate interferon (Interferon, IFN) is to activate a series of expression of interferon-induced property genes, to resist and remove virus.Wherein, I Type interferon (Type I IFN, including IFN-α and IFN-β) be body resist virus infection key innate immune cells because Son, classical Antiviral Mechanism mainly pass through the expression of up-regulation I type interferon.
However dependent on the treatment of the clinical disease of viral infection of interferon, there are certain drawbacks.For example, interferon is controlled Treatment is only effective to certain types of disease of viral infection, and effective percentage is to be improved；Can be due to there is interferon resistance in body Influence therapeutic effect；Interferon therapy application can also generate the side effects such as flu syndrome, alopecia；Interferon can induce certainly The generation (Kretschmer S etc., Curr Opin Immunol, 2017,49:96-102) of body immunity disease and inflammation.
Therefore, seek the innate immunity/interferon response dependent/non-dependent regulation virus infection new target drone and method for facing Bed treatment disease of viral infection is significant.
Specific test in the disclosure shows that itaconic acid can promote virus infection and duplication in cell or Mice Body, and And the raising of this process simultaneous I type interferon expression amount.This demonstrate that itaconic acid does not pass through inhibition interferon expression This classical antiviral natural immunologic mechanism promotes virus infection, i.e., the effect that itaconic acid promotees virus infection is naturally to exempt from Epidemic disease/interferon response is non-dependent.This, which is found to be, understands the non-dependent cytophylaxis mode of the innate immunity and removing pathogen Mechanism provides new visual angle.
Using itaconic acid mortifier treat disease of viral infection and/or its associated disease compare other innate immunitys or IFN dependent drug has advantage outstanding, to have huge application potential.These advantages may include but be not limited to:
It (1), can't when the strategy using targeted inhibition itaconic acid and its relevant metabolic pathway carries out antiviral therapy Enhance the generation of interferon, this itself can exempt to avoid inflammatory caused by interferon excess and/or innate immunity activation The generation of epidemic disease disease.So that relative to other innate immunitys or IFN dependent drug, with itaconic acid and its metabolic pathway Have for the drug target of clinical treatment compared with strong security；
(2) inhibition of itaconic acid and its metabolic pathway/blocking treatment method is particularly suitable for innate immune function and is damaged/defect Or the object that interferon therapy is invalid or ineffective.The object of/defect impaired for innate immune function, using the innate immunity Or IFN dependent drug carries out treating the expression that can not effectively activate the downstream antiviral effect factor, to can not effectively send out It waves resistance and removes the effect of virus.And itaconic acid promotes the effect of virus infection not by the regulation innate immunity and interferon Response realizes, therefore, the method for targeted inhibition itaconic acid and its metabolic pathway can/defect impaired in innate immune function or Antiviral effect still is played in the case that interferon therapy is invalid/ineffective, this is infectious for prevention and control and treatment poison The clinical application of disease has popularity applicability and significance.
The mortifier of the itaconic acid (including its metabolic pathway) of the disclosure is particularly well suited to lack with the innate immunity as a result, Fall into/impaired and/or interferon anti-reflecting virus treatment of infection is invalid or ineffective or is also easy to produce the object of side effect, such as suffer from itself Immunity disease or the object for being susceptible to suffer from autoimmune disease, as IFN insufficiency or too drastic object (such as suffer from inflammatory bowel Disease, rheumatoid arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, Chronic gastrointestinal diseases object).
Product and its application
The disclosure additionally provides a kind of product, can be drug, pharmaceutical composition or medicine box, wherein containing a effective amount of Disclosed itaconic acid mortifier, and acceptable carrier pharmaceutically or in immunology.As used herein, term " active material " Or " active constituent " is used interchangeably, and refers to itaconic acid mortifier.
The mortifier and product of the disclosure can be used for preventing and/or treating virus infection or associated disease and/or Illness.Infection is DNA virus and/or picornavirus infection, such as the infection caused by one or more viruses selected from the group below: Herpes simplex virus, vesicular stomatitis virus, influenza virus, encephalomyocarditis virus, sendai virus, hepatitis type B virus, adenopathy Poison, poxvirus, small DNA virus, adeno-associated virus.
The mortifier and product of the disclosure can be used for preventing and/or treating disease relevant to virus infection and/or symptom One of to be selected from the group or a variety of: pathology damage caused by virus infection；The cells following viral infection factor (such as interferon) It generates insufficient or excessive；Endotoxic shock or death；The inflammatory damage of organ；Multiple organ failure, such as the organ It is selected from: liver, spleen, brain, kidney, heart, lung, stomach, intestines；Chronic inflammation disease caused by virus infection (such as autoimmunity Property disease such as inflammatory bowel disease, rheumatoid arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, chronic gastrointestinal disease Disease).
The mortifier or product of the disclosure can be used for different objects, preferably mammalian object, such as people, pet, family Poultry.Also, the mortifier or product of the disclosure have advantage when preventing or treating the object with innate immunity defect, such as With innate immunity defect/impaired and/or interferon anti-reflecting virus treatment of infection is invalid or ineffective or is also easy to produce side effect Object such as suffers from autoimmune disease or is susceptible to suffer from the object of autoimmune disease, such as IFN insufficiency or too drastic object (as suffered from inflammatory bowel disease, rheumatoid arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, chronic gastrointestinal disease The object of disease).
Therefore, itaconic acid mortifier or product comprising the mortifier are additionally provided in the disclosure have in treatment naturally exempt from The application in disease of viral infection and/or associated disease and/or illness in the object of epidemic disease defect.In the disclosure also Providing itaconic acid mortifier or the product comprising the mortifier has the disease in IFN insufficiency or too drastic object in treatment Application in malicious infectious diseases and/or associated disease and/or illness.Itaconic acid mortifier is additionally provided in the disclosure Or the product comprising the mortifier disease of viral infection and/or phase therewith in object of the treatment with autoimmune disease The disease of pass and/or the application in illness.
As used herein, term " containing " or " comprising " include "comprising", " substantially by ... constitute " and " by ... constitute ".As used herein, term " pharmaceutically acceptable " ingredient is suitable for people and/or animal and without excessively not Good reaction (such as toxicity, stimulation and allergy), that is, there is the substance of reasonable benefit/risk ratio.As used herein, term " effective quantity ", which refers to, to generate function or amount that is active and being received by people and/or animal to people and/or animal.
As used herein, term " pharmaceutically acceptable carrier " refers to for preventing and/or the carrier of Therapeutic Administration, packet Include various excipient and diluent.The term refers to medicament carriers some in this way: it themselves is not necessary active constituent, and There is no excessive toxicity after application.Suitably carrier is well known to those of ordinary skill in the art, in " Remington drug section Learn " it can find about medicine in (Remington ' s Pharmaceutical Sciences, Mack Pub.Co., N.J.1991) Acceptable excipient discusses fully on.
Pharmaceutically acceptable carrier can contain liquid in the composition, such as water, salt water, glycerol and ethyl alcohol.In addition, these There is likely to be complementary substances in carrier, such as filler, disintegrating agent, lubricant, glidant, effervescent agent, wetting agent or cream Agent, corrigent, pH buffer substance etc..In general, these substances can be formulated in nontoxic, inert and pharmaceutically acceptable In aqueous carrier medium, wherein pH is usually about 5-8, preferably, pH is about 6-8.
Active material in the product of the disclosure accounts for 0.001~99.9wt% of composition total weight；Preferably composition 1~95wt% of total weight is more preferably 5~90wt%, more preferable 10~80wt%.Surplus can be pharmaceutically acceptable load The substances such as body and other additives.
As used herein, term " unit dosage forms " refers in order to convenient to take, and the product of the disclosure is prepared into single clothes With required dosage form, including but not limited to various solid formulations (such as tablet), liquid agent, capsule, sustained release agent.
In other embodiments of the disclosure, the product is unit dosage form or multi-form, and wherein active material Content be 0.001~2000mg/ agent, 0.01~1500mg/ agent, 0.1~1000mg/ agent.In other implementations of the disclosure In mode, the composition of 1~6 dose of disclosure, such as 1~3 dose or 1 dose are applied daily.
It should be understood that the effective dose of mortifier used can change with the severity of to be administered or treatment object.Tool Body situation determines according to the individual instances (such as object weight, age, physical condition, required effect reached) of object, this In the range of skilled practitioners may determine that.
The product of the disclosure can be solid-state (such as granule, tablet, freeze-dried powder, suppository, capsule, sublingual lozenge) or liquid (such as oral solution) or other suitable shapes.Administration route can be used but be not limited to according to the type of mortifier: oral, injection (such as direct naked DNA or protein injection method, liposome DNA or protein injection method), gold coating particle bombardment, Breeding unsoundness bacterium carries Plasmid DNA method, replication defective adenoviral carries protein coded by target DNA method or target gene, Electroporation, pulmonary administration, oral administration, cutaneous penetration, is administered in tumor nasal-cavity administration.
In addition, also containing other active matters for improving and treating disease of viral infection in the product of the disclosure Matter, other active materials include but is not limited to: clinical common antibiotics, including beta-lactam (penicillins and head Spore bacteriums), aminoglycosides, Tetracyclines, chloromycetin, macrolide, antifungal antibiotic, treating tuberculosis class antibiosis Element；Clinic common antiviral drugs (tricyclic amines, pyrophosphoric acid class, albumen enzyme inhibitor, nucleoside medicine and interferon, antisense Oligonucleotides etc.)；The common immunosuppressor of clinic (including glucocorticoid, cyclophosphamide, chloroquine, Ciclosporin A, Thunder God Rattan, Chinese materia medica preparation, anti-TNF monoclonal antibody).
Also, the mortifier or product of the disclosure each other can be with use in conjunction, can also be with other medicines and treatment hand Duan Lianhe, the treatment for disease of viral infection.
Drug screening method
Screening is additionally provided in the disclosure using itaconic acid as the method for the viral infection resisting drug candidate of target spot.
In some embodiments, screening includes as follows using itaconic acid as the method for the viral infection resisting drug candidate of target spot Step:
(A) infected cell, tissue or animal are handled with candidate substances；
(B) detect the cell, itaconic acid in tissue or animal, its metabolic precursor thereof (such as itaconate, aconitate), Itaconic acid metabolite, itaconic acid generate the level or activity of required enzyme (such as IRG1 or OGDH)；And
(C) if level or activity detected is lower than the respective horizontal or activity before candidate substances processing or lower than normal The infection of respective horizontal or activity and the cell, tissue or animal in control is controlled or is suppressed, then shows described Candidate substances may be as by inhibiting itaconic acid come the drug of viral infection resisting.
In some embodiments, the method still further comprise detection the candidate substances processing in or later, The variation of IFN level in cell, tissue or the animal being infected.If IFN level is without significant changes or significant decrease, and disease Poison infection is controlled or is inhibited, then shows that the safety of the candidate substances is higher, in particular for innate immunity imbalance or IFN Safety for insufficiency or autoimmune disease object is higher.
Embodiment
Combined with specific embodiments below, the disclosure is further described.It should be understood that these embodiments are merely to illustrate the disclosure Rather than it limits the scope of the present disclosure.Those skilled in the art can make modification appropriate, variation to the disclosure, these modifications All within the scope of the present disclosure with variation.
In the following examples, the experimental methods for specific conditions are not specified, the conventional method in this field can be used, such as join Examine " Molecular Cloning:A Laboratory guide " (third edition, New York, CSH Press, New York:Cold Spring Harbor Laboratory Press, 1989) or according to condition proposed by supplier.The sequencing approach of DNA is that this field is normal The method of rule can also provide test by commercial company.
Unless otherwise stated, otherwise percentage and number are calculated by weight.Unless otherwise defined, as used herein all Professional and scientific terms have the same meanings as commonly understood by one of ordinary skill in the art.In addition, any similar or equal to described content Deng method and material all can be applied in method of disclosure.Preferred implement methods and materials described in the text only present a demonstration it With.
Embodiment 1: the mouse primary peritoneal macrophage underpants health acid content of virus infection significantly increases
Mouse primary peritoneal macrophage be isolated from intraperitoneal injection 2ml 3% thioglycolate salt (according to % be m/v ratio, Dissolved using distilled water, carry out high pressure sterilization processing later) C57BL/6 mouse (6-8 week old, male) after 3 days, it is incubated at In the DMEM in high glucose culture medium of endotoxin-free containing 10% fetal calf serum (FBS).
Using vesicular stomatitis virus (Vesicular Stomatitis Virus, VSV are purchased from ATCC) with the agent of MOI=1 Amount stimulation cell different time points.Cell is collected, 0.9% physiology salt being pre-chilled with 4 DEG C is washed three times, and -80 DEG C are added later in advance Cold 80% methanol-water containing the D4- succinic acid (Succinic acid-d4, as internal controls) that concentration is 10 μ g/ml Solution (v/v).Sample is collected, ultrasound is centrifuged 15 minutes under the conditions of 16,000g, 4 DEG C after 30 seconds and removes insoluble fragment.
Join in 8060 liquid chromatography-tandem mass spectrometry of LC-MS/MS of the positive/negative ionization mode equipped with electrospray ionisation source Targeting metabolism group is carried out to the cell sample of preparation on instrument to detect to obtain the absolute content of itaconic acid.Mass Spectrometer Method data use Shimadzu LabSolutions Postrun Analysis (version 5.89) software carries out processing analysis, target metabolic The integration peak area of object is normalized with internal controls to carry out sxemiquantitative.
D4- succinic acid is purchased from Sigma Aldrich；8060 liquid chromatography-tandem mass spectrometry of LC-MS/MS connection instrument is purchased from Shimadzu Corporation company；Shimadzu LabSolutions Postrun Analysis(version 5.89) Analysis software is developed by Shimadzu company.Each numerical point in figure indicates that independent biochemical repeats.
As the result is shown: virus infection can be such that mouse primary peritoneal macrophage underpants health acid content significantly increases.
Result prompt: the level of metabolin itaconic acid in cell, and virus infection can significantly be raised in virus infection Degree is higher, and itaconic acid content is more, this illustrates that itaconic acid can play regulating and controlling effect during virus infection.It can pass through Whether the content of kit or metabolism group means detection itaconic acid is infected judging body and gradient of infection.
Embodiment 2: itaconic acid can promote the virus replication in mouse primary peritoneal macrophage in a dose-dependent manner
Mouse primary peritoneal macrophage is isolated from the mouse after intraperitoneal injection thioglycolate salt 3 days, is incubated at and contains (with embodiment 1) in the DMEM in high glucose culture medium of the endotoxin-free of 10% fetal calf serum (FBS), and trained overnight in 37 DEG C of incubators It supports.
Change within second day fresh complete medium into, and can be in body with the existing document report of various concentration shown in Fig. 2 The cell for being inside converted into itaconic acid can absorb derivatives from itaconic acid dimethyl itaconate (Dimethyl itaconate, DMI) (LampropoulouV etc., Cell Metab.2016；24 (1): 158-66) (Fig. 2A) or 4- octyl itaconic acid (4-Octyl Itaconate, OI) (Mills EL etc., Nature, 2018,556 (7699): 113-117) (Fig. 2 B) respective pretreatment cell 3 Hour.Above-mentioned derivatives from itaconic acid needs to be configured to the fresh concentrate that concentration is 200mM, and root with DMSO before every use Use is diluted according to needs.The DMSO of equivalent volumes is then added into culture medium in the cell of control group (dosage 0).Later, Cell stimulates 8 hours with VSV virus, and extracts RNA and carry out reverse transcription.VSV virus replication level, which is pressed, uses quantitative fluorescent PCR (qPCR) method is tested and analyzed.
The used absorbable derivatives from itaconic acid of cell include: dimethyl itaconate (Dimethyl itaconate, DMI) it is purchased from Sigma company；4- octyl itaconic acid (4-Octyl itaconate, OI) is purchased from MedChemExpress company.
Detect the qPCR primer sequence of VSV virus replication sheet:
As the result is shown: can significantly increase mouse original with two kinds of derivatives from itaconic acid DMI and OI respective pretreatment cell For the virus replication level in peritoneal macrophage, and promote the effect of virus replication that there is dose dependent.
Result prompt: itaconic acid has the function of remarkably promoting virus replication, can be by for itaconic acid upstream generation The siRNA for thanking to enzyme presses down with CRISPR/Cas9 gene editing product, specific antibody or itaconic acid metabolic pathway inhibitor The content of metabolin itaconic acid processed, to inhibit virus infection to protect body from virus infection.
Embodiment 3: itaconic acid has the function of promoting virus replication in Mice Body
According to the ratio of 50mg/kg weight, 4- octyl itaconic acid (4-Octyl is injected to mouse peritoneal within 2 hours in advance Itaconate, OI) (being dissolved in 40% (v/v) cyclodextrin by PBS configuration), later according to 5 × 107PFU/g weight is to mouse VSV virus is injected intravenously to infect 18 hours.Liver, spleen and the lungs for collecting mouse later extract RNA and carry out reverse transcription, Using virus replication level in the qPCR method detection internal organs in embodiment 2.
The used absorbable derivatives from itaconic acid 4- octyl itaconic acid of cell (4-Octyl itaconate, OI) purchase From in MedChemExpress company.
As the result is shown: itaconic acid class compound injection to mouse peritoneal, which is carried out pretreatment, can remarkably promote mice organs Middle virus replication level.
Should the result shows that: itaconic acid have the function of in Mice Body promote virus replication, indication targeted inhibition itaconic acid The strategy for generating or playing a role is used clinically for preventing and/or treating the potential using value of disease of viral infection.Tool It, can be by siRNA for itaconic acid upstream metabolic enzyme and CRISPR/Cas9 gene editing product, special for body Property antibody or itaconic acid metabolic pathway inhibitor inhibit the content of metabolin itaconic acid, to inhibit virus infection to protect machine Body is from virus infection.
Embodiment 4: itaconic acid promotes Mice Body inner virus to infect by the non-dependent mode of interferon
Using the method in embodiment 3, according to the ratio of 50mg/kg weight, 2 hours in advance pungent to mouse peritoneal injection 4- Base itaconic acid (4-Octyl itaconate, OI) (is dissolved in 40% (v/v) cyclodextrin by PBS configuration), later according to 5 × 107PFU/g weight infects 18 hours (with embodiment 3) to mouse mainline VSV virus.The serum of mouse is collected, is used Antiviral relevant I type interferon (Type I in ELISA kit (being purchased from R&D company) detection mice serum Interferon) the protein level of IFN-β (Fig. 4 A) or IFN-α (Fig. 4 B).
As the result is shown: compared to control group mice, in the case of virus stimulation, itaconic acid class compound is pre-processed so that mouse IFN-β (Fig. 4 A) or IFN-α (Fig. 4 B) protein level in serum significantly increase.
The result proves: itaconic acid promotes the effect of virus infection and duplication in Mice Body, disease-resistant not by inhibition The generation of toxic effect factor interferon and realize.This can be to avoid using targeted inhibition itaconic acid progress antiviral therapy strategy When, since interferon generates generation that is excessive and leading to autoimmune disease, it was demonstrated that using itaconic acid as the medicine of clinical treatment Object target spot has compared with strong security.
Embodiment 5: it strikes low metabolic enzyme OGDH expression and can lower cell underpants health acid content and carry out suppressing virus replication
A: the jamming effectiveness of the siRNA (siRNA) of targeting mouse metabolism enzyme OGDH
Mouse primary peritoneal macrophage is isolated from the mouse (with embodiment 1) after intraperitoneal injection thioglycolate salt 3 days, It is incubated in the DMEM in high glucose culture medium of the endotoxin-free containing 10% fetal calf serum (FBS).
SiRNAs for interference mice OGDH gene uses TransIT-siQUEST Transfection Reagent Transfection reagent (being purchased from Mirus company) is transfected into primary peritoneal macrophage (12 hole cell culture according to operating instruction respectively The transfection parameters of plate: 100 μ l, TransIT-siQUEST Transfection Reagent of serum free medium 3 μ l, 25nM 2.8 μ l of siRNA of final concentration).After 48 hours, use Western blot detection OGDH protein expression level to verify three Target the jamming effectiveness (Fig. 5 A) of the siRNA of mouse OGDH.
It is purchased from Genepharma company for the siRNAs of mouse OGDH gene, specific siRNAs sequence is as follows:
B: metabolin itaconic acid and suitable crow in mouse primary peritoneal macrophage can be lowered by striking low metabolic enzyme OGDH expression The content of head acid
Using TransIT-siQUEST Transfection Reagent transfection reagent (being purchased from Mirus company) according to behaviour The siRNA that will shine and target respectively mouse OGDH is explained (using the 3 siRNA mixing for targeting mouse OGDH in embodiment 5A Using) be transfected into primary peritoneal macrophage (transfection parameters of 12 porocyte culture plates: 100 μ l of serum free medium, 2.8 μ l of siRNA of 3 μ l, 25nM final concentration of TransIT-siQUEST Transfection Reagent).Interference 48 hours Afterwards, using VSV virus (MOI=1 is purchased from ATCC) 6 hours of infection cell.0.9% physiology salt that cell is pre-chilled with 4 DEG C is washed Three times, D4- succinic acid (the Succinic acid-d4, as interior for being 10 μ g/ml containing concentration of -80 DEG C of pre-coolings is added later Ministerial standard product) 80% methanol aqueous solution (v/v).Sample is collected, ultrasound is centrifuged 15 points under the conditions of 16,000g, 4 DEG C after 30 seconds Clock removes insoluble fragment.
Metabolism group is targeted in 8060 liquid chromatogram of LC-MS/MS-of the positive/negative ionization mode equipped with electrospray ionisation source It is detected on tandem mass spectrum connection instrument.Mass Spectrometer Method data use Shimadzu LabSolutions Postrun Analysis (version 5.89) software carries out processing analysis, and the integration peak area of target metabolite is normalized with internal controls to carry out Sxemiquantitative.
D4- succinic acid is purchased from Sigma Aldrich；8060 liquid chromatography-tandem mass spectrometry of LC-MS/MS connection instrument comes from Shimadzu Corporation company；Shimadzu LabSolutions Postrun Analysis(version 5.89) Analysis software is developed by Shimadzu company.
C: low OGDH expression is struck in mouse primary peritoneal macrophage can inhibit virus replication (left figure), which does not depend on Regulation (right figure) to interferon response
With embodiment 5A, after interfering OGDH to express 48 hours in Turnover of Mouse Peritoneal Macrophages, using VSV virus (MOI= 1, it is purchased from ATCC) infection cell 8 hours, RNA is extracted later and carries out reverse transcription, using real time fluorescence quantifying PCR method (qPCR) VSV viral RNA levels (Fig. 5 C is left) and the level of IFN-β mRNA are detected (Fig. 5 C is right).
The qPCR primer sequence of VSV viral RNA levels is detected with embodiment 2；Detect the qPCR of mouse IFN-β mRNA level in-site Primer sequence is as follows:
Test result and discussion
As the result is shown: the albumen table of OGDH in low cell can be effectively struck using the siRNAs for mouse metabolism enzyme OGDH Up to (Fig. 5 A).And interfere OGDH that can lower the content (figure of itaconic acid and itaconic acid upstream metabolin aconitate in cell 5B), thus inhibit the expression (Fig. 5 C) of duplication amount and IFN-β mRNA viral in cell.
The result proves: by the strategy of targeted inhibition itaconic acid, such as the table by interfering or inhibiting metabolic enzyme OGDH It reaches and/or activity is to reduce itaconic acid upstream metabolin aconitate content, the duplication of virus can be inhibited to remove viral sense Dye, and this effect is independent of interferon response.The inhibition itaconic acid content dominance of strategies is to cause excessive Interferon generates, and avoids the damage and side effect to body.
It is incorporated herein by reference in all documents that the disclosure refers to, it is independent just as each document It is incorporated as with reference to such.In addition, it should also be understood that, those skilled in the art can after having read the above-mentioned teaching content of the disclosure To make various changes or modifications to the disclosure, such equivalent forms equally fall within model defined by the application the appended claims It encloses.
Sequence table
<110>Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences
<120>application of the itaconic acid related substances in diagnosis, prevention and treatment disease of viral infection
<130> 196629 1CNCN
<160> 14
<170> PatentIn version 3.3
<210> 1
<211> 19
<212> DNA
<213>artificial sequence
<400> 1
acggcgtact tccagatgg 19
<210> 2
<211> 19
<212> DNA
<213>artificial sequence
<400> 2
ctcggttcaa gatccaggt 19
<210> 3
<211> 21
<212> RNA
<213>artificial sequence
<400> 3
ggacuugugc ugcuaaguut t 21
<210> 4
<211> 21
<212> RNA
<213>artificial sequence
<400> 4
aacuuagcag cacaagucct t 21
<210> 5
<211> 21
<212> RNA
<213>artificial sequence
<400> 5
cccgcugaca uuaucucaut t 21
<210> 6
<211> 21
<212> RNA
<213>artificial sequence
<400> 6
augagauaau gucagcgggt t 21
<210> 7
<211> 21
<212> RNA
<213>artificial sequence
<400> 7
gggaaagacc aaagcugaat t 21
<210> 8
<211> 21
<212> RNA
<213>artificial sequence
<400> 8
uucagcuuug gucuuuccct t 21
<210> 9
<211> 21
<212> DNA
<213>artificial sequence
<400> 9
cagctccaag aaaggacgaa c 21
<210> 10
<211> 21
<212> DNA
<213>artificial sequence
<400> 10
ggcagtgtaa ctcttctgca t 21
<210> 11
<211> 3117
<212> RNA
<213>mouse (Mus musculus)
<400> 11
atgtttcatt taaggacttg tgctgctaag ttaaggccat tgacagcctc ccagactgtt 60
aagacatttt cacaaaacaa accagcagca attaggacgt ttcaacagat tcggtgctat 120
tctgcacctg tagctgctga accatttctt agtgggacta gttcgaacta tgtggaggaa 180
atgtactgtg cctggttgga gaatcccaaa agtgtacata agtcatggga catttttttc 240
cgaaacacca atgctggagc cccaccgggc actgcctacc agagccccct ttccctgagt 300
cgaagctccc tggctaccat ggcccatgca cagtccctgg tggaagcaca acctaacgtc 360
gacaaactcg tggaggacca cttggcggtg cagtctctca tcagggcata tcagatacga 420
gggcaccatg tagcacagct ggaccccctg gggattttgg atgctgatct ggactcctcc 480
gtgcccgctg acattatctc atccacagac aaacttgatc ttgcagtttt caaggaacga 540
cttcgaatgc taacagtagg agggttctat ggcctacacg agtctgacct tgacaaggtc 600
ttccacttac ccaccaccac tttcatcggg ggacaggagc cagcacttcc tcttcgggag 660
atcatccgtc ggctggagat ggcctactgc cagcacattg gtgtggagtt catgttcatt 720
aatgatttgg aacaatgcca gtggatccga cagaagtttg agacccctgg aatcatgcag 780
ttcaccaatg aggagaagcg gaccttgctg gccaggcttg tacgatccac caggtttgag 840
gagttcctac agcgaaagtg gtcctcggag aagcgttttg gtctggaagg ctgtgaggtg 900
ctgatccctg ccctcaagac aatcattgat atgtcaagtg caaatggagt ggactatgtg 960
atcatgggga tgccacacag aggacggctg aacgtgcttg caaatgtcat caggaaggag 1020
ctggagcaaa tattctgtca gtttgactca aagctggagg cagctgatga gggttctggg 1080
gacatgaagt accacctggg catgtatcac cgcaggatca accgtgtgac cgacagaaac 1140
atcactttgt ccttggtggc taacccttcc catctagagg ctgctgaccc tgtcgtgatg 1200
ggaaagacca aagctgaaca gttctactgt ggagacactg aagggaaaaa ggtgatgtct 1260
atcctgctgc atggggatgc tgcttttgct ggccagggca tcgtatatga gaccttccat 1320
ctcagcgact tgccgtccta cacaacccat ggcactgttc atgtggttgt caacaaccag 1380
attggcttca ccacagaccc tcggatggcc cgctcctctc cctaccccac tgatgtggcc 1440
cgagtggtga atgcccccat tttccatgtc aactcagatg accctgaagc tgtcatgtat 1500
gtatgcaagg tggcagctga gtggagaaac accttccaca aggatgttgt agttgatctg 1560
gtgtgttatc gacgaaatgg ccacaatgag atggacgaac ctatgtttac acagccactc 1620
atgtacaagc agatccgcaa gcagaagcct gtactgcaga agtatgcaga attgctagtc 1680
tcccagggtg tcgtcaatca gcctgagtac gaggaggaaa tctccaagta tgataagatc 1740
tgtgaggaag catttaccag atccaaagat gagaagatct tgcacatcaa gcactggctg 1800
gattccccct ggcctggctt tttcaccctg gatggacagc ccaggagcat gacctgcccc 1860
tccactggcc tggaggagga tgtcttgttc cacattggaa aggtggccag ctctgtacct 1920
gtggagaact ttactatcca tggagggctg agccggatct tgaagacccg cagagagctt 1980
gtgacgaacc ggactgtgga ctgggccctg gcagagtaca tggcatttgg ctcactgctg 2040
aaggaaggca tccatgtgcg gctgagtggc caggatgtgg agcggggcac cttcagccat 2100
cgccaccatg tgctccatga tcagaatgtt gacaaaagaa cctgcatccc catgaaccac 2160
ctttggccaa atcaggcccc ttacactgta tgcaacagct cgctgtctga gtacggtgtc 2220
ctgggctttg agctgggctt tgccatggct agccctaatg ctctggttct ctgggaggcc 2280
cagtttggtg acttcaacaa catggcacag tgcatcattg accagttcat ctgcccagga 2340
caggcaaagt gggtgcggca gaatggcatt gtgctcctgc tgcctcatgg catggaaggc 2400
atgggtcccg agcattcctc tgcccgccca gagcggtttc tgcagatgtg caatgatgac 2460
ccagatgtcc tgcctgacct gcaggaagaa aactttgaca tcaatcagct ctatgactgc 2520
aactggattg ttgtcaactg ttccacccct ggcaacttct tccatgtgct gcgacgacag 2580
atcttgctgc ccttccggaa gccgttaatc gtcttcactc ccaaatccct cctgcgccac 2640
cctgaggcaa gaactagctt tgacgagatg ctgccaggaa cgcacttcca gcgtgtgatc 2700
ccagaaaatg gacctgcagc tcaggaccca cacaaagtca agagacttct cttctgcact 2760
gggaaggtgt actatgacct cacccgagag cgcaaagcca ggaacatgga ggaggaggtg 2820
gctattacaa ggattgagca gctatcacca ttcccctttg acctcctgct gaaagaggca 2880
cagaagtatc ccaatgctga gctggcctgg tgccaggaag agcacaagaa ccaaggctac 2940
tatgactatg tcaagccaag acttcgtacc accattgacc gtgctaagcc tgtctggtat 3000
gctggccgag acccggcagc tgctccagcc actggcaaca agaaaacaca cctgacagag 3060
ctgcagcgct ttctggacac agcctttgac ctggacgcat tcaagaaatt ctcttag 3117
<210> 12
<211> 1038
<212> PRT
<213>mouse (Mus musculus)
<400> 12
Met Phe His Leu Arg Thr Cys Ala Ala Lys Leu Arg Pro Leu Thr Ala
1 5 10 15
Ser Gln Thr Val Lys Thr Phe Ser Gln Asn Lys Pro Ala Ala Ile Arg
20 25 30
Thr Phe Gln Gln Ile Arg Cys Tyr Ser Ala Pro Val Ala Ala Glu Pro
35 40 45
Phe Leu Ser Gly Thr Ser Ser Asn Tyr Val Glu Glu Met Tyr Cys Ala
50 55 60
Trp Leu Glu Asn Pro Lys Ser Val His Lys Ser Trp Asp Ile Phe Phe
65 70 75 80
Arg Asn Thr Asn Ala Gly Ala Pro Pro Gly Thr Ala Tyr Gln Ser Pro
85 90 95
Leu Ser Leu Ser Arg Ser Ser Leu Ala Thr Met Ala His Ala Gln Ser
100 105 110
Leu Val Glu Ala Gln Pro Asn Val Asp Lys Leu Val Glu Asp His Leu
115 120 125
Ala Val Gln Ser Leu Ile Arg Ala Tyr Gln Ile Arg Gly His His Val
130 135 140
Ala Gln Leu Asp Pro Leu Gly Ile Leu Asp Ala Asp Leu Asp Ser Ser
145 150 155 160
Val Pro Ala Asp Ile Ile Ser Ser Thr Asp Lys Leu Asp Leu Ala Val
165 170 175
Phe Lys Glu Arg Leu Arg Met Leu Thr Val Gly Gly Phe Tyr Gly Leu
180 185 190
His Glu Ser Asp Leu Asp Lys Val Phe His Leu Pro Thr Thr Thr Phe
195 200 205
Ile Gly Gly Gln Glu Pro Ala Leu Pro Leu Arg Glu Ile Ile Arg Arg
210 215 220
Leu Glu Met Ala Tyr Cys Gln His Ile Gly Val Glu Phe Met Phe Ile
225 230 235 240
Asn Asp Leu Glu Gln Cys Gln Trp Ile Arg Gln Lys Phe Glu Thr Pro
245 250 255
Gly Ile Met Gln Phe Thr Asn Glu Glu Lys Arg Thr Leu Leu Ala Arg
260 265 270
Leu Val Arg Ser Thr Arg Phe Glu Glu Phe Leu Gln Arg Lys Trp Ser
275 280 285
Ser Glu Lys Arg Phe Gly Leu Glu Gly Cys Glu Val Leu Ile Pro Ala
290 295 300
Leu Lys Thr Ile Ile Asp Met Ser Ser Ala Asn Gly Val Asp Tyr Val
305 310 315 320
Ile Met Gly Met Pro His Arg Gly Arg Leu Asn Val Leu Ala Asn Val
325 330 335
Ile Arg Lys Glu Leu Glu Gln Ile Phe Cys Gln Phe Asp Ser Lys Leu
340 345 350
Glu Ala Ala Asp Glu Gly Ser Gly Asp Met Lys Tyr His Leu Gly Met
355 360 365
Tyr His Arg Arg Ile Asn Arg Val Thr Asp Arg Asn Ile Thr Leu Ser
370 375 380
Leu Val Ala Asn Pro Ser His Leu Glu Ala Ala Asp Pro Val Val Met
385 390 395 400
Gly Lys Thr Lys Ala Glu Gln Phe Tyr Cys Gly Asp Thr Glu Gly Lys
405 410 415
Lys Val Met Ser Ile Leu Leu His Gly Asp Ala Ala Phe Ala Gly Gln
420 425 430
Gly Ile Val Tyr Glu Thr Phe His Leu Ser Asp Leu Pro Ser Tyr Thr
435 440 445
Thr His Gly Thr Val His Val Val Val Asn Asn Gln Ile Gly Phe Thr
450 455 460
Thr Asp Pro Arg Met Ala Arg Ser Ser Pro Tyr Pro Thr Asp Val Ala
465 470 475 480
Arg Val Val Asn Ala Pro Ile Phe His Val Asn Ser Asp Asp Pro Glu
485 490 495
Ala Val Met Tyr Val Cys Lys Val Ala Ala Glu Trp Arg Asn Thr Phe
500 505 510
His Lys Asp Val Val Val Asp Leu Val Cys Tyr Arg Arg Asn Gly His
515 520 525
Asn Glu Met Asp Glu Pro Met Phe Thr Gln Pro Leu Met Tyr Lys Gln
530 535 540
Ile Arg Lys Gln Lys Pro Val Leu Gln Lys Tyr Ala Glu Leu Leu Val
545 550 555 560
Ser Gln Gly Val Val Asn Gln Pro Glu Tyr Glu Glu Glu Ile Ser Lys
565 570 575
Tyr Asp Lys Ile Cys Glu Glu Ala Phe Thr Arg Ser Lys Asp Glu Lys
580 585 590
Ile Leu His Ile Lys His Trp Leu Asp Ser Pro Trp Pro Gly Phe Phe
595 600 605
Thr Leu Asp Gly Gln Pro Arg Ser Met Thr Cys Pro Ser Thr Gly Leu
610 615 620
Glu Glu Asp Val Leu Phe His Ile Gly Lys Val Ala Ser Ser Val Pro
625 630 635 640
Val Glu Asn Phe Thr Ile His Gly Gly Leu Ser Arg Ile Leu Lys Thr
645 650 655
Arg Arg Glu Leu Val Thr Asn Arg Thr Val Asp Trp Ala Leu Ala Glu
660 665 670
Tyr Met Ala Phe Gly Ser Leu Leu Lys Glu Gly Ile His Val Arg Leu
675 680 685
Ser Gly Gln Asp Val Glu Arg Gly Thr Phe Ser His Arg His His Val
690 695 700
Leu His Asp Gln Asn Val Asp Lys Arg Thr Cys Ile Pro Met Asn His
705 710 715 720
Leu Trp Pro Asn Gln Ala Pro Tyr Thr Val Cys Asn Ser Ser Leu Ser
725 730 735
Glu Tyr Gly Val Leu Gly Phe Glu Leu Gly Phe Ala Met Ala Ser Pro
740 745 750
Asn Ala Leu Val Leu Trp Glu Ala Gln Phe Gly Asp Phe Asn Asn Met
755 760 765
Ala Gln Cys Ile Ile Asp Gln Phe Ile Cys Pro Gly Gln Ala Lys Trp
770 775 780
Val Arg Gln Asn Gly Ile Val Leu Leu Leu Pro His Gly Met Glu Gly
785 790 795 800
Met Gly Pro Glu His Ser Ser Ala Arg Pro Glu Arg Phe Leu Gln Met
805 810 815
Cys Asn Asp Asp Pro Asp Val Leu Pro Asp Leu Gln Glu Glu Asn Phe
820 825 830
Asp Ile Asn Gln Leu Tyr Asp Cys Asn Trp Ile Val Val Asn Cys Ser
835 840 845
Thr Pro Gly Asn Phe Phe His Val Leu Arg Arg Gln Ile Leu Leu Pro
850 855 860
Phe Arg Lys Pro Leu Ile Val Phe Thr Pro Lys Ser Leu Leu Arg His
865 870 875 880
Pro Glu Ala Arg Thr Ser Phe Asp Glu Met Leu Pro Gly Thr His Phe
885 890 895
Gln Arg Val Ile Pro Glu Asn Gly Pro Ala Ala Gln Asp Pro His Lys
900 905 910
Val Lys Arg Leu Leu Phe Cys Thr Gly Lys Val Tyr Tyr Asp Leu Thr
915 920 925
Arg Glu Arg Lys Ala Arg Asn Met Glu Glu Glu Val Ala Ile Thr Arg
930 935 940
Ile Glu Gln Leu Ser Pro Phe Pro Phe Asp Leu Leu Leu Lys Glu Ala
945 950 955 960
Gln Lys Tyr Pro Asn Ala Glu Leu Ala Trp Cys Gln Glu Glu His Lys
965 970 975
Asn Gln Gly Tyr Tyr Asp Tyr Val Lys Pro Arg Leu Arg Thr Thr Ile
980 985 990
Asp Arg Ala Lys Pro Val Trp Tyr Ala Gly Arg Asp Pro Ala Ala Ala
995 1000 1005
Pro Ala Thr Gly Asn Lys Lys Thr His Leu Thr Glu Leu Gln Arg
1010 1015 1020
Phe Leu Asp Thr Ala Phe Asp Leu Asp Ala Phe Lys Lys Phe Ser
1025 1030 1035
<210> 13
<211> 3105
<212> RNA
<213>homo sapiens (Homo sapiens)
<400> 13
atgtttcatt taaggacttg tgctgctaag ttgaggccat tgacggcttc ccagactgtt 60
aagacatttt cacaaaacag accagcagca gctaggacat ttcaacagat tcggtgctat 120
tctgcacctg ttgctgctga gccctttctc agtgggacta gttcgaacta tgtggaggag 180
atgtactgtg cttggctgga aaaccccaaa agtgtacata agtcatggga catttttttt 240
cgcaacacga atgccggagc cccaccgggc actgcctacc agagtcccct tcccctgagc 300
cgaggctccc tggctgctgt ggcccatgca cagtccctgg tagaagcaca gcccaacgtg 360
gacaagctcg tggaggacca cctggcagtg cagtcgctca tcagggcata tcaggtcagg 420
ggtcaccaca ttgcaaaact tgatcctctc ggaattagtt gtgtaaattt tgatgatgct 480
ccagtaactg tttcttcaaa cgtggatctt gcagttttca aggaacgact tcgaatgcta 540
acagtaggag ggttctatgg cctggatgag tctgacctcg acaaggtctt ccacttgccc 600
accaccactt tcatcggggg acaggaatca gcacttcctc tgcgggagat catccgtcgg 660
ctggagatgg cctactgcca gcatattggg gtggagttca tgttcatcaa tgacctggag 720
cagtgccagt ggatccggca gaagtttgag acccctggga tcatgcagtt cacaaatgag 780
gagaaacgga ccctgctggc caggcttgtg cggtccacca ggtttgagga gttcctacag 840
cggaagtggt cctctgagaa gcgctttggt ctagaaggct gcgaggtact gatccctgcc 900
ctcaagacca tcattgacaa gtctagtgag aatggcgtgg actacgtgat catgggcatg 960
ccacacagag ggcggctgaa cgtgcttgca aatgtcatca ggaaggagct ggaacagatc 1020
ttctgtcaat tcgattcaaa gctggaggca gctgatgagg gctccggaga tgtgaagtac 1080
cacctgggca tgtatcaccg caggatcaat cgtgtcaccg acaggaacat taccttgtcc 1140
ttggtggcca acccttccca ccttgaggcc gctgaccccg tggtgatggg caagaccaaa 1200
gccgaacagt tttactgtgg cgacactgaa gggaaaaagg tcatgtccat cctgttgcat 1260
ggggatgctg catttgctgg ccagggcatt gtgtacgaga ccttccacct cagcgacctg 1320
ccatcctaca caactcatgg caccgtgcac gtggtcgtca acaaccagat cggcttcacc 1380
accgaccctc ggatggcccg ctcctccccc taccccactg acgtggcccg agtggtgaat 1440
gcccccattt tccacgtgaa ctcagatgac cccgaggctg tcatgtacgt gtgcaaagtg 1500
gcggccgagt ggaggagcac cttccacaag gacgtggttg tcgatttggt gtgttaccgg 1560
cgcaacggcc acaacgagat ggatgagccc atgttcacgc agccgctcat gtacaagcag 1620
atccgcaagc agaagcctgt gttacagaag tacgctgagc tgctggtgtc gcagggtgtg 1680
gtcaaccagc ctgagtatga ggaggaaatt tccaagtatg ataagatctg tgaggaagct 1740
tttgccagat ctaaagatga gaagatcttg cacattaagc actggctgga ctctccctgg 1800
cctggcttct tcaccctgga cgggcagccc aggagcatgt cctgcccctc cacgggtctg 1860
acggaggata ttctgacaca catcgggaat gtggctagtt ctgtgcctgt ggaaaacttt 1920
actattcatg gagggctgag ccggatcttg aagactcgtg gggaaatggt gaagaaccgg 1980
actgtggact gggctctagc ggagtacatg gcgtttggct cgctcctgaa ggagggcatc 2040
cacattcggc tgagcggcca ggacgtggag cggggcacat tcagccaccg ccaccatgtg 2100
ctccatgacc agaatgtgga caagagaacc tgcatcccca tgaaccatct ctggcccaat 2160
caggccccct atactgtgtg caacagctca ctgtctgagt acggcgtgct gggctttgag 2220
ctgggcttcg ccatggccag tcctaatgcc ctggtcctct gggaagccca atttggtgac 2280
ttccacaaca cggcccagtg tatcatcgac cagttcatct gcccgggaca agccaagtgg 2340
gtgcggcaga atggcatcgt gttgctgctg ccccatggca tggagggcat gggtccagaa 2400
cattcctccg cccgcccaga gcggttcttg cagatgtgca acgatgaccc agatgtcctg 2460
ccagacctta aagaagccaa cttcgacatc aatcagctat atgactgcaa ttgggttgtt 2520
gtcaactgct ccactcctgg caacttcttc cacgtgctac gacgccagat cctgctgcca 2580
ttccggaagc cgttaattat cttcaccccc aaatccctgt tgcgccaccc cgaggccaga 2640
tccagctttg atgagatgct tccaggaacc cacttccagc gggtgatccc agaagatggc 2700
cctgcagctc agaacccaga aaatgtcaaa aggcttctct tctgcaccgg caaagtgtat 2760
tatgacctca cccgggagcg caaagcacgc gacatggtgg ggcaggtggc catcacaagg 2820
attgagcagc tgtcgccatt cccctttgac ctcctgctga aggaggtgca gaagtacccc 2880
aatgctgagc tggcctggtg ccaggaggag cacaagaacc aaggctacta tgactacgtg 2940
aagccaagac ttcggaccac catcagccgc gccaagcccg tctggtatgc cggccgggac 3000
ccagcggctg ctccagccac cggcaacaag aagacccacc tgacggagct gcagcgcctc 3060
ctggacacgg ccttcgacct ggacgtcttc aagaacttct cgtag 3105
<210> 14
<211> 1034
<212> PRT
<213>homo sapiens (Homo sapiens)
<400> 14
Met Phe His Leu Arg Thr Cys Ala Ala Lys Leu Arg Pro Leu Thr Ala
1 5 10 15
Ser Gln Thr Val Lys Thr Phe Ser Gln Asn Arg Pro Ala Ala Ala Arg
20 25 30
Thr Phe Gln Gln Ile Arg Cys Tyr Ser Ala Pro Val Ala Ala Glu Pro
35 40 45
Phe Leu Ser Gly Thr Ser Ser Asn Tyr Val Glu Glu Met Tyr Cys Ala
50 55 60
Trp Leu Glu Asn Pro Lys Ser Val His Lys Ser Trp Asp Ile Phe Phe
65 70 75 80
Arg Asn Thr Asn Ala Gly Ala Pro Pro Gly Thr Ala Tyr Gln Ser Pro
85 90 95
Leu Pro Leu Ser Arg Gly Ser Leu Ala Ala Val Ala His Ala Gln Ser
100 105 110
Leu Val Glu Ala Gln Pro Asn Val Asp Lys Leu Val Glu Asp His Leu
115 120 125
Ala Val Gln Ser Leu Ile Arg Ala Tyr Gln Val Arg Gly His His Ile
130 135 140
Ala Lys Leu Asp Pro Leu Gly Ile Ser Cys Val Asn Phe Asp Asp Ala
145 150 155 160
Pro Val Thr Val Ser Ser Asn Val Asp Leu Ala Val Phe Lys Glu Arg
165 170 175
Leu Arg Met Leu Thr Val Gly Gly Phe Tyr Gly Leu Asp Glu Ser Asp
180 185 190
Leu Asp Lys Val Phe His Leu Pro Thr Thr Thr Phe Ile Gly Gly Gln
195 200 205
Glu Ser Ala Leu Pro Leu Arg Glu Ile Ile Arg Arg Leu Glu Met Ala
210 215 220
Tyr Cys Gln His Ile Gly Val Glu Phe Met Phe Ile Asn Asp Leu Glu
225 230 235 240
Gln Cys Gln Trp Ile Arg Gln Lys Phe Glu Thr Pro Gly Ile Met Gln
245 250 255
Phe Thr Asn Glu Glu Lys Arg Thr Leu Leu Ala Arg Leu Val Arg Ser
260 265 270
Thr Arg Phe Glu Glu Phe Leu Gln Arg Lys Trp Ser Ser Glu Lys Arg
275 280 285
Phe Gly Leu Glu Gly Cys Glu Val Leu Ile Pro Ala Leu Lys Thr Ile
290 295 300
Ile Asp Lys Ser Ser Glu Asn Gly Val Asp Tyr Val Ile Met Gly Met
305 310 315 320
Pro His Arg Gly Arg Leu Asn Val Leu Ala Asn Val Ile Arg Lys Glu
325 330 335
Leu Glu Gln Ile Phe Cys Gln Phe Asp Ser Lys Leu Glu Ala Ala Asp
340 345 350
Glu Gly Ser Gly Asp Val Lys Tyr His Leu Gly Met Tyr His Arg Arg
355 360 365
Ile Asn Arg Val Thr Asp Arg Asn Ile Thr Leu Ser Leu Val Ala Asn
370 375 380
Pro Ser His Leu Glu Ala Ala Asp Pro Val Val Met Gly Lys Thr Lys
385 390 395 400
Ala Glu Gln Phe Tyr Cys Gly Asp Thr Glu Gly Lys Lys Val Met Ser
405 410 415
Ile Leu Leu His Gly Asp Ala Ala Phe Ala Gly Gln Gly Ile Val Tyr
420 425 430
Glu Thr Phe His Leu Ser Asp Leu Pro Ser Tyr Thr Thr His Gly Thr
435 440 445
Val His Val Val Val Asn Asn Gln Ile Gly Phe Thr Thr Asp Pro Arg
450 455 460
Met Ala Arg Ser Ser Pro Tyr Pro Thr Asp Val Ala Arg Val Val Asn
465 470 475 480
Ala Pro Ile Phe His Val Asn Ser Asp Asp Pro Glu Ala Val Met Tyr
485 490 495
Val Cys Lys Val Ala Ala Glu Trp Arg Ser Thr Phe His Lys Asp Val
500 505 510
Val Val Asp Leu Val Cys Tyr Arg Arg Asn Gly His Asn Glu Met Asp
515 520 525
Glu Pro Met Phe Thr Gln Pro Leu Met Tyr Lys Gln Ile Arg Lys Gln
530 535 540
Lys Pro Val Leu Gln Lys Tyr Ala Glu Leu Leu Val Ser Gln Gly Val
545 550 555 560
Val Asn Gln Pro Glu Tyr Glu Glu Glu Ile Ser Lys Tyr Asp Lys Ile
565 570 575
Cys Glu Glu Ala Phe Ala Arg Ser Lys Asp Glu Lys Ile Leu His Ile
580 585 590
Lys His Trp Leu Asp Ser Pro Trp Pro Gly Phe Phe Thr Leu Asp Gly
595 600 605
Gln Pro Arg Ser Met Ser Cys Pro Ser Thr Gly Leu Thr Glu Asp Ile
610 615 620
Leu Thr His Ile Gly Asn Val Ala Ser Ser Val Pro Val Glu Asn Phe
625 630 635 640
Thr Ile His Gly Gly Leu Ser Arg Ile Leu Lys Thr Arg Gly Glu Met
645 650 655
Val Lys Asn Arg Thr Val Asp Trp Ala Leu Ala Glu Tyr Met Ala Phe
660 665 670
Gly Ser Leu Leu Lys Glu Gly Ile His Ile Arg Leu Ser Gly Gln Asp
675 680 685
Val Glu Arg Gly Thr Phe Ser His Arg His His Val Leu His Asp Gln
690 695 700
Asn Val Asp Lys Arg Thr Cys Ile Pro Met Asn His Leu Trp Pro Asn
705 710 715 720
Gln Ala Pro Tyr Thr Val Cys Asn Ser Ser Leu Ser Glu Tyr Gly Val
725 730 735
Leu Gly Phe Glu Leu Gly Phe Ala Met Ala Ser Pro Asn Ala Leu Val
740 745 750
Leu Trp Glu Ala Gln Phe Gly Asp Phe His Asn Thr Ala Gln Cys Ile
755 760 765
Ile Asp Gln Phe Ile Cys Pro Gly Gln Ala Lys Trp Val Arg Gln Asn
770 775 780
Gly Ile Val Leu Leu Leu Pro His Gly Met Glu Gly Met Gly Pro Glu
785 790 795 800
His Ser Ser Ala Arg Pro Glu Arg Phe Leu Gln Met Cys Asn Asp Asp
805 810 815
Pro Asp Val Leu Pro Asp Leu Lys Glu Ala Asn Phe Asp Ile Asn Gln
820 825 830
Leu Tyr Asp Cys Asn Trp Val Val Val Asn Cys Ser Thr Pro Gly Asn
835 840 845
Phe Phe His Val Leu Arg Arg Gln Ile Leu Leu Pro Phe Arg Lys Pro
850 855 860
Leu Ile Ile Phe Thr Pro Lys Ser Leu Leu Arg His Pro Glu Ala Arg
865 870 875 880
Ser Ser Phe Asp Glu Met Leu Pro Gly Thr His Phe Gln Arg Val Ile
885 890 895
Pro Glu Asp Gly Pro Ala Ala Gln Asn Pro Glu Asn Val Lys Arg Leu
900 905 910
Leu Phe Cys Thr Gly Lys Val Tyr Tyr Asp Leu Thr Arg Glu Arg Lys
915 920 925
Ala Arg Asp Met Val Gly Gln Val Ala Ile Thr Arg Ile Glu Gln Leu
930 935 940
Ser Pro Phe Pro Phe Asp Leu Leu Leu Lys Glu Val Gln Lys Tyr Pro
945 950 955 960
Asn Ala Glu Leu Ala Trp Cys Gln Glu Glu His Lys Asn Gln Gly Tyr
965 970 975
Tyr Asp Tyr Val Lys Pro Arg Leu Arg Thr Thr Ile Ser Arg Ala Lys
980 985 990
Pro Val Trp Tyr Ala Gly Arg Asp Pro Ala Ala Ala Pro Ala Thr Gly
995 1000 1005
Asn Lys Lys Thr His Leu Thr Glu Leu Gln Arg Leu Leu Asp Thr
1010 1015 1020
Ala Phe Asp Leu Asp Val Phe Lys Asn Phe Ser
1025 1030

Claims (10)

1. itaconic acid mortifier is in preparation for disease of viral infection and/or and virus infection in prevention and/or treatment object Purposes in the product of relevant disease and/or symptom.
2. purposes as described in claim 1, which is characterized in that the itaconic acid mortifier is selected from the inhibition for following substance Object: enzyme needed for itaconic acid, itaconic acid metabolic precursor thereof (such as aconitate), metabolism generate itaconic acid (such as aconitate is de- Carboxylic acid IRG1, ketoglurate dehydrogenase OGDH)；
In some embodiments, the itaconic acid mortifier is selected from: the upstream metabolic enzyme generated for itaconic acid or its precursor Antibody (such as antibody for IRG1 or OGDH), siRNA (such as in SEQ ID NO:3-8 it is any shown in be directed to OGDH SiRNA), miRNA, antisense oligonucleotides, CRISPR/Cas9 gene editing product, antagonist, blocking agent, inhibit itaconic acid Or the gene of enzyme (such as OGDH or IRG1) needed for its preceding body function and/or active compound, downward metabolism generation itaconic acid Or the promoter element and/or expression vector of mRNA or protein expression level.
3. purposes as described in claim 1, which is characterized in that it is described infection be DNA virus and/or picornavirus infection, such as The infection caused by one or more viruses selected from the group below: herpes simplex virus, vesicular stomatitis virus, influenza virus, brain Myocarditis virus, sendai virus, hepatitis type B virus, adenovirus, poxvirus, small DNA virus, adeno-associated virus；And/or
The disease relevant to virus infection and/or symptom are one of to be selected from the group or a variety of: caused by virus infection Pathology damage；The cells following viral infection factor (such as interferon) generates insufficient or excessive；Endotoxic shock or death；Organ Inflammatory damage；Multiple organ failure, such as the organ are selected from: liver, spleen, brain, kidney, heart, lung, stomach, intestines；Virus sense Chronic inflammation disease caused by contaminating (such as autoimmune disease such as inflammatory bowel disease, rheumatoid arthritis, systemic red Yabbi sore, chronic nephritis, tuberculosis, Chronic gastrointestinal diseases).
4. purposes as described in claim 1, which is characterized in that the object is mammal, such as people, pet, domestic animal, example Such as there is innate immunity defect/impaired and/or interferon anti-reflecting virus treatment of infection is invalid or ineffective or is also easy to produce side effect Object, such as with autoimmune disease or the object of autoimmune disease is susceptible to suffer from, such as IFN insufficiency or too drastic object (as suffered from inflammatory bowel disease, rheumatoid arthritis, systemic loupus erythematosus, chronic nephritis, tuberculosis, chronic gastrointestinal disease The object of disease).
5. purposes as described in claim 1, which is characterized in that the product is pharmaceutical composition or medicine box, such as its form Suitable for by being selected from the group the pharmaceutical composition or medicine box that mode is given: oral, injection (such as direct naked DNA or protein note Penetrate method, liposome DNA or protein injection method), gold coating particle bombardment, breeding unsoundness bacterium carry Plasmid DNA Protein coded by method, replication defective adenoviral carrying target DNA method or target gene, electroporation, nasal-cavity administration, lung give Medicine, cutaneous penetration, is administered in tumor oral administration；And/or
The product also include for prevent and/or treat disease of viral infection and/or disease relevant to virus infection and/ Or other substances of symptom, such as one of be selected from the group or a variety of: clinical common antibiotics, including beta-lactam are (green Mycin class and cephalosporins), aminoglycosides, Tetracyclines, chloromycetin, macrolide, antifungal antibiotic, resistive connection Core class antibiotic；Clinic common antiviral drugs (tricyclic amines, pyrophosphoric acid class, albumen enzyme inhibitor, nucleoside medicine and interference Element, antisense oligonu-cleotides etc.)；The common immunosuppressor of clinic (including glucocorticoid, cyclophosphamide, chloroquine, cyclosporin A, tripterygium wilfordii, Chinese materia medica preparation, anti-TNF monoclonal antibody).
6. a kind of pharmaceutical composition or medicine box, it includes:
(A) the itaconic acid mortifier of prevention and/or therapeutically effective amount；
(B) acceptable carrier or excipient pharmaceutically or in immunology；
(C) optionally, one or more preventions and/or treatment disease of viral infection or illness relevant to virus infection And/or other active materials of symptom.
7. the reagent of levels of substance is preparing judging viral infection disease or its sense in detection itaconic acid and/or its metabolic pathway Application in the kit of dye degree.
8. the use as claimed in claim 7, wherein substance is selected from the itaconic acid metabolic pathway: itaconic acid metabolic precursor thereof (such as itaconate, aconitate), itaconic acid metabolite, itaconic acid generate required enzyme (such as IRG1 or OGDH).
9. a kind of kit comprising: detection is obtained from levels of substance in itaconic acid in the sample of object and/or its metabolic pathway Reagent.
10. a kind of method of drug candidate of screening by inhibition itaconic acid come viral infection resisting comprising:
(A) infected cell, tissue or animal are handled with candidate substances；
(B) itaconic acid, its metabolic precursor thereof (such as itaconate, aconitate), clothing health in the detection cell, tissue or animal Acid metabolic product, itaconic acid generate the level or activity of required enzyme (such as IRG1 or OGDH)；And
(C) if level or activity detected is lower than the respective horizontal or activity before candidate substances processing or is lower than normal control In respective horizontal or the infection of activity and the cell, tissue or animal be controlled or be suppressed, then show the candidate Substance may be as by inhibiting itaconic acid come the drug of viral infection resisting.
CN201910769226.0A 2019-08-20 2019-08-20 Application of itaconic acid related substance in diagnosis and prevention of virus infectious diseases Active CN110507823B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910769226.0A CN110507823B (en) 2019-08-20 2019-08-20 Application of itaconic acid related substance in diagnosis and prevention of virus infectious diseases

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910769226.0A CN110507823B (en) 2019-08-20 2019-08-20 Application of itaconic acid related substance in diagnosis and prevention of virus infectious diseases

Publications (2)

Publication Number Publication Date
CN110507823A true CN110507823A (en) 2019-11-29
CN110507823B CN110507823B (en) 2021-08-24

Family

ID=68627031

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910769226.0A Active CN110507823B (en) 2019-08-20 2019-08-20 Application of itaconic acid related substance in diagnosis and prevention of virus infectious diseases

Country Status (1)

Country Link
CN (1) CN110507823B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115400113A (en) * 2022-08-05 2022-11-29 复旦大学附属中山医院 Application of 4-octyl itaconic acid in preparing medicine for preventing or treating ovarian cancer
CN115443131A (en) * 2020-04-17 2022-12-06 纳布里瓦治疗有限责任公司 Therapeutic use of pleuromutilins
CN116139117A (en) * 2022-05-31 2023-05-23 西南医科大学附属医院 Application of itaconic acid and derivative thereof in preventing and treating cardiotoxicity induced by doxorubicin
WO2024126742A1 (en) * 2022-12-16 2024-06-20 Universite De Tours Aconitate for the treatment of a viral lung infection

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115443131A (en) * 2020-04-17 2022-12-06 纳布里瓦治疗有限责任公司 Therapeutic use of pleuromutilins
CN116139117A (en) * 2022-05-31 2023-05-23 西南医科大学附属医院 Application of itaconic acid and derivative thereof in preventing and treating cardiotoxicity induced by doxorubicin
CN115400113A (en) * 2022-08-05 2022-11-29 复旦大学附属中山医院 Application of 4-octyl itaconic acid in preparing medicine for preventing or treating ovarian cancer
WO2024126742A1 (en) * 2022-12-16 2024-06-20 Universite De Tours Aconitate for the treatment of a viral lung infection

Also Published As

Publication number Publication date
CN110507823B (en) 2021-08-24

Similar Documents

Publication Publication Date Title
CN110507823A (en) Application of the itaconic acid related substances in diagnosis, prevention and treatment disease of viral infection
US10045961B2 (en) Use of sting agonists to treat chronic hepatitis B virus infection
Widagdo et al. MERS-coronavirus: From discovery to intervention
Yue et al. Effects of triterpenes from Ganoderma lucidum on protein expression profile of HeLa cells
CN112930392B (en) Probiotics for upper respiratory tract infections, stress, anxiety, memory and cognitive dysfunction and aging
Zhou et al. Metformin regulates tight junction of intestinal epithelial cells via MLCK-MLC signaling pathway.
CN110420331A (en) Application of the ALKBH5 mortifier in treatment disease of viral infection
Hu et al. Emerging role of the cGAS-STING signaling pathway in autoimmune diseases: Biologic function, mechanisms and clinical prospection
CN110101705A (en) The anti-viral uses of BET family protein inhibitor
Zhang et al. Macrophage transcriptional responses following in vitro infection with a highly virulent African swine fever virus isolate
Caza et al. The zinc finger protein Mig1 regulates mitochondrial function and azole drug susceptibility in the pathogenic fungus Cryptococcus neoformans
Wang et al. Axl deficiency promotes the neuroinvasion of Japanese encephalitis virus by enhancing IL-1α production from pyroptotic macrophages
Jeswin et al. Proteomic analysis by iTRAQ in red claw crayfish, Cherax quadricarinatus, hematopoietic tissue cells post white spot syndrome virus infection
CN101969993A (en) Synthetic replikin peptides against pathogenic infection of invertebrates in aquaculture
CN1605027A (en) Methods and compositions for treatment of central nervous system disorders
Liu et al. Madecassoside ameliorates lipopolysaccharide-induced neurotoxicity in rats by activating the Nrf2-HO-1 pathway
Marini et al. HIV‐1 matrix protein p17 binds to monocytes and selectively stimulates MCP‐1 secretion: role of transcriptional factor AP‐1
Wang et al. Muscovy duck reovirus p10. 8 protein induces ER stress and apoptosis through the Bip/IRE1/XBP1 pathway
Oliveira et al. Chicken cGAS senses fowlpox virus infection and regulates macrophage effector functions
Liu et al. SLC1A3 facilitates Newcastle disease virus replication by regulating glutamine catabolism
Abdolmaleky et al. Bioactive nutraceuticals oligo-lactic acid and fermented soy extract alleviate cognitive decline in mice in part via anti-neuroinflammation and modulation of gut microbiota
Wang et al. Molecular characterization of diphthamide biosynthesis protein 7 in Marsupenaeus japonicus and its role in white spot syndrome virus infection
Amin et al. Gut-joint axis: Oral Probiotic ameliorates Osteoarthritis
Yan et al. WSV152 induces apoptosis and promotes viral replication in Litopenaeus vannamei
Du et al. Alisol B 23-acetate broadly inhibits coronavirus through blocking virus entry and suppresses proinflammatory T cells responses for the treatment of COVID-19

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