CN109481684A - Application using Optineurin as target spot in preparation prevention and treatment autoimmune disease drug - Google Patents
Application using Optineurin as target spot in preparation prevention and treatment autoimmune disease drug Download PDFInfo
- Publication number
- CN109481684A CN109481684A CN201811568313.1A CN201811568313A CN109481684A CN 109481684 A CN109481684 A CN 109481684A CN 201811568313 A CN201811568313 A CN 201811568313A CN 109481684 A CN109481684 A CN 109481684A
- Authority
- CN
- China
- Prior art keywords
- optineurin
- optn
- autoimmune disease
- target spot
- preparation prevention
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
It is application of the target spot in preparation prevention and treatment autoimmune disease drug that the present invention, which is provided using Optineurin, the drug include antibody in conjunction with OPTN protein-specific, Optineurin gene specific siRNA, sh-OPTN gland related diseases, lead to the Optineurin micromolecular inhibitors of steroid derivatives shown in formula (I).Research of the present invention has shown that promote CD4 by inhibiting Optn gene expression that can inhibit maturing dendritic cell+T cell inhibits CD4 to suppression scorching Th2 and Treg polarization+T cell polarizes to proinflammatory Th1 and Th17, to inhibit self immune system, plays the effect for the treatment of autoimmune disease such as multiple sclerosis and rheumatoid arthritis.Present invention discloses new role of the Optn gene in immunocyte and immune system, and the drug or other treatment means of prevention and/or treatment autoimmune disease are prepared using Optn gene or albumen as shot design.
Description
Technical field
The present invention relates to pharmaceutical technology fields, specifically, being related to using Optineurin being target spot in preparation prevention and treatment itself
Application in immunity disease drug.
Background technique
Optic nerve albumen Optineurin (OPTN) is encoded by Optn gene, and the hair of autophagy is participated in mainly as autophagy receptor
It is raw.The function of the albumen is not yet fully elucidated at present, studies have reported that it is green with amyotrophic lateral sclerosis, primary open-angle
Light eye is closely related；Optn gene unconventionality expression in Crow grace enteritis and paget's disease of bone patient is had been reported that in immunology,
It may participate in the occurrence and development of immunity disease.Autoimmune disease is that immune system occurs to be immunized instead to body itself composition
The disease answered and caused, main pathologic process are as follows: the Dendritic Cells secretion inflammatory factor and chemotactic factor (CF) of activation, and will
Autoantigen presentation gives original CD4+T cell (Naive CD4+T), make Naive CD4+T cell activation is extended and is divided into
Different subtype, including causing scorching Th1, Th17 cell, Th2 the and Treg cell of He Yiyan finally starts immune response.Currently,
Existing 100 various autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis, lupus erythematosus, it has also become cardiovascular
Third class principal disease after disease and tumour.These diseases are often chronic protracted course of disease, clinically be there is no effective
Prevention and therapeutic agent are eventually led to disabled or dead.Therefore, need to research and develop a kind of efficient diagnosis and prevention and treatment autoimmune disease
Drug or other treatment means.
Summary of the invention
It is target spot in preparation prevention and treatment autoimmune disease drug that the purpose of the application, which is to provide using Optineurin,
It include siRNA, sh-OPTN adenopathy of antibody in conjunction with OPTN protein-specific, Optn gene specific using, the drug
The Optn micromolecular inhibitor of steroid derivatives shown in poison, logical formula (I).
Described be target spot with Optineurin (Optn) (Gene ID:10133) includes being subtracted using gene knockout, clpp gene
Either chemicals reduce the expression of Optn or make Optn silencing.
The autoimmune disease includes multiple sclerosis, rheumatoid arthritis, it is not limited here.
Wherein:
The positive-sense strand of siRNA sequence as shown in Seq ID No.1, antisense strand sequence as shown in Seq ID No.2,
Sh-OPTN adenoviral plasmid sequence is as shown in Seq ID No.3:
Seq ID No.1:5 '-GCCUGUUGUUUGAGAUGCAAA-3 '
Seq ID No.2:5 '-UUUGCAUCUCAAACAACAGGC-3 '
Seq ID No.3:TTTGAGGAGCTTTCGGCCTGGACAGAGAA.
The general structure of steroid derivatives shown in logical formula (I) are as follows:
In particular compound shown in formula (II):
The Optn gene knockout of the application can inhibit maturing dendritic cell, promote CD4+T cell to the scorching Th2 of suppression and
Treg polarization, inhibits CD4+T cell polarizes to proinflammatory Th1 and Th17, to inhibit self immune system.
Micromolecular inhibitor provided by the present application, especially compound shown in (II), confirm the little molecules in inhibiting after study
Agent can specificity inhibition Optn gene expression, achieve the purpose that treat autoimmune disease.
The present invention is experiments have shown that in multiple sclerosis (MS), rheumatoid arthritis (RA) patient peripheral's blood monocyte
Expression is apparently higher than normal healthy controls in OptnmRNA level；It knocks out Optn and can significantly inhibit EAE (MS animal model), (RA is dynamic by CIA
Object model) mouse severity.The present invention is demonstrated simultaneously by inhibiting Optn gene expression, can inhibit Dendritic Cells
Maturation promotes CD4+T cell inhibits CD4 to suppression scorching Th2 and Treg polarization+T cell polarizes to proinflammatory Th1 and Th17, into
And weaken adaptive immunity reaction, play the role for the treatment of autoimmune disease, and have developed new autoimmune accordingly
Disease treatment mode: Optn gene specific siRNA, sh-OPTN virus, micromolecular inhibitor and with OPTN protein-specific knot
The antibody of conjunction.The application is using Optn as the drug of shot design treatment and/prevention of autoimmune diseases.Optn is disclosed to exist
New role in immunocyte and immune system, and new therapeutic targets and effectively new are provided for prevention and treatment autoimmune disease
Medicine.
Detailed description of the invention
Fig. 1 is the present invention by qRT-PCR method detection multiple sclerosis (MS), outside rheumatoid arthritis (RA) patient
The mRNA level in-site of all blood monocyte (PBMC) Optn.
Fig. 2 is using Optn-/-Mouse detects the relaxation effect to EAE and CIA mice clinical symptoms.
Fig. 3 is using WT and Optn-/-Mouse extracts and cultivates derived from bone marrow dendritic cells (BMDC), flow cytometry
Detect influence of the Optn to BMDC maturation.
Fig. 4 is using WT and Optn-/-The lymphoid tissue airflow classification of mouse simultaneously cultivates Naive CD4+T cell, streaming are thin
Born of the same parents' art detects Optn to Naive CD4+T cell is polarized to Th1, Th2, Th17, the influence of Treg cell.
Fig. 5 is detection micromolecular inhibitor C to the effect of Optn mRNA level in-site and its to EAE and CIA mice clinical symptoms
Relaxation effect.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this
It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip
Part, or according to the normal condition proposed by manufacturer.
Embodiment 1:
Collect MS, RA clinical sample peripheral blood each 30.Peripheral blood mononuclear cells is obtained using Ficoll gradient separations
(PBMC), using the mRNA level in-site of qRT-PCR detection Optn.Referring to Fig. 1, the mRNA level in-site of Optn in the PBMC of MS, RA patient
Higher than normal healthy controls (Healthy Donor, HD).
Embodiment 2:
Using WT and Optn-/-C57BL/6 adult female mice constructs EAE model.Specific EAE modeling method is as follows: will
MOG35-55It is dissolved in sterile PBS, and is prepared by mixing into emulsion with the complete Freund's adjuvant containing tubercle bacillus.In mouse neck and small
Emulsion, while tail vein injection pertussis toxin is subcutaneously injected in mouse or so hind leg.The tail vein injection one hundred days again of modeling second day
Cough toxin.EAE simulates clinical multiple Sclerosis Symptoms standards of grading are as follows: 0 point, asymptomatic；1 point, flaccid tail；2 points, tail
Inability and part of limb inability；3 points, side hind limb paralysis；4 points；Two sides hind limb paralysis；It is 5 points, dying or dead.Work as from modeling
It plays (being denoted as Day 0), carries out the marking of EAE symptom daily.And in disease end, collect mouse spinal cord sample, frozen section into
Row LFB dyeing.A referring to fig. 2, B, Optn-/-EAE mouse gives a mark compared with WT mice clinical and significantly reduces, and LFB dyeing display
Optn-/-EAE myelin is more complete.
Using WT and Optn-/-C57BL/6 adult male mice constructs CIA model.Specific CIA modeling method is as follows: by chicken
II Collagenase Type is dissolved in sterile PBS, and is prepared by mixing into emulsion with complete Freund's adjuvant.In mouse tail root emulsion for injection,
It is denoted as day 0, and the emulsion for injection again in immune latter 21 days.CIA symptom generally starts to occur after secondary immunity, rheumatoid arthrosis
Scorching arthroncus degree is scored with 0~4 grade, and 0 point: without arthroncus；1 point: small toe joint mild swelling；2 points: small toe joint
With plantar redness；3 points: ankle-joint sufficient pawl swelling below；4 points: whole arthroncus including ankle-joint.Scoring 1 point with
On be just defined as rheumatoid arthritis.From after secondary immunity, the marking of CIA symptom is carried out daily.And in disease end, collect small
Mouse joint, paraffin section carry out HE dyeing.C referring to fig. 2, D, Optn-/-CIA mouse gives a mark compared with WT mice clinical and significantly reduces, and
HE dyeing display, which knocks out Optn, can alleviate joint injury.
Embodiment 3:
Extract WT and Optn-/-LPS is given after the derived from bone marrow dendritic cells (BMDC) of mouse, collects cell after 24 hours
CD11C and CD80 is contaminated altogether, and MHC I or MHC II detect dendritic cell maturation situation.Referring to Fig. 3 A-C, knocks out Optn and inhibit dendron
Cell maturation.
Embodiment 4:
Airflow classification extracts WT and Optn-/-The naive CD4 of mouse+T, in a.Th1 polarization culture medium；B.Th17 polarization
Culture medium；C.Th2 polarization culture medium；D.Treg polarization culture medium is cultivated.Referring to fig. 4, it knocks out Optn and inhibits naive CD4+T is polarized to Th1 and Th17 cell, promotes naive CD4+T is polarized to Th2 and Treg cell.
Embodiment 5:
The micromolecular inhibitor C that various concentration is acted on 293T cell, collects cell after 24 hours, qRT-PCR is investigated
Inhibiting effect of the micromolecular inhibitor C to Optn mRNA level in-site.It is in dose-dependent inhibition referring to Fig. 5 A, micromolecular inhibitor C
Optn mRNA level in-site.
EAE model is constructed using WT C57BL/6 adult female mice, is divided into solvent control group and experimental group.It is sent out in disease
When raw, CMC-Na is given in the daily stomach-filling of solvent group, and various dose micromolecular inhibitor C is given in the daily stomach-filling of experimental group.Referring to figure
5B, micromolecular inhibitor C reduce the marking of EAE clinic in dose dependent.
CIA model is constructed using WT C57BL/6 adult male mice, is divided into solvent control group and experimental group.It is sent out in disease
When raw, CMC-Na is given in the daily stomach-filling of solvent group, and various dose micromolecular inhibitor C is given in the daily stomach-filling of experimental group.Referring to figure
5C, micromolecular inhibitor C reduce the marking of CIA clinic in dose dependent.
Sequence table
<110>Zhejiang University
<120>application using Optineurin as target spot in preparation prevention and treatment autoimmune disease drug
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA/RNA
<213>artificial sequence (Unknown)
<400> 1
gccuguuguu ugagaugcaa a 21
<210> 2
<211> 21
<212> DNA/RNA
<213>artificial sequence (Unknown)
<400> 2
uuugcaucuc aaacaacagg c 21
<210> 3
<211> 29
<212> DNA/RNA
<213>artificial sequence (Unknown)
<400> 3
tttgaggagc tttcggcctg gacagagaa 29
Claims (6)
1. the application using Optineurin as target spot in preparation prevention and treatment autoimmune disease drug, which is characterized in that described
Drug includes that antibody, siRNA, sh-OPTN gland of Optineurin gene specific in conjunction with OPTN protein-specific are related
The Optineurin micromolecular inhibitor of steroid derivatives shown in virus, logical formula (I).
2. according to claim 1 using Optineurin as answering in the preparation prevention and treatment autoimmune disease drug of target spot
With, which is characterized in that it is described that using Optineurin as target spot, including using, gene knockout, clpp gene subtract or chemicals reduce
The expression of Optineurin makes Optineurin silencing.
3. the answering in preparation prevention and treatment autoimmune disease drug according to claim 1 using Optineurin as target spot
It include multiple sclerosis with, which is characterized in that the autoimmune disease, rheumatoid arthritis, it is not limited here.
4. the answering in preparation prevention and treatment autoimmune disease drug according to claim 1 using Optineurin as target spot
With, which is characterized in that the positive-sense strand of siRNA sequence as shown in Seq ID No.1, antisense strand is as shown in Seq ID No.2
Sequence, sh-OPTN virus particle sequence is as shown in Seq ID No.3:
Seq ID No.1:5 '-gccuguuguuugagaugcaaa-3 '
Seq ID No.2:5 '-uuugcaucucaaacaacaggc-3 '
Seq ID No.3:tttgaggagctttcggcctggacagagaa.
5. the answering in preparation prevention and treatment autoimmune disease drug according to claim 1 using Optineurin as target spot
With, which is characterized in that the general structure of steroid derivatives shown in logical formula (I) are as follows:
6. the answering in preparation prevention and treatment autoimmune disease drug according to claim 5 using Optineurin as target spot
With, which is characterized in that the steroid derivatives are compound shown in formula (II):
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811568313.1A CN109481684B (en) | 2018-12-21 | 2018-12-21 | Application of Optineurin as target in preparation of medicine for preventing and treating autoimmune diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811568313.1A CN109481684B (en) | 2018-12-21 | 2018-12-21 | Application of Optineurin as target in preparation of medicine for preventing and treating autoimmune diseases |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109481684A true CN109481684A (en) | 2019-03-19 |
| CN109481684B CN109481684B (en) | 2020-10-27 |
Family
ID=65711263
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811568313.1A Active CN109481684B (en) | 2018-12-21 | 2018-12-21 | Application of Optineurin as target in preparation of medicine for preventing and treating autoimmune diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109481684B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110327332A (en) * | 2019-07-06 | 2019-10-15 | 河北医科大学第二医院 | Amlexanox is alleviating application and its experimental method in EAE morbidity |
| CN111733233A (en) * | 2020-05-09 | 2020-10-02 | 中国人民解放军陆军军医大学 | Application of RORCE2 gene as drug target in preparation of drugs for treating Th17 cell-related diseases |
| CN113318230A (en) * | 2021-06-11 | 2021-08-31 | 上海交通大学医学院附属第九人民医院 | Application of Optineurin in diagnosis and treatment of ocular melanoma |
| CN113896757A (en) * | 2020-07-07 | 2022-01-07 | 中国科学院上海药物研究所 | Pentacyclic triterpene carbon glycoside compounds, and preparation method and application thereof |
-
2018
- 2018-12-21 CN CN201811568313.1A patent/CN109481684B/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110327332A (en) * | 2019-07-06 | 2019-10-15 | 河北医科大学第二医院 | Amlexanox is alleviating application and its experimental method in EAE morbidity |
| CN111733233A (en) * | 2020-05-09 | 2020-10-02 | 中国人民解放军陆军军医大学 | Application of RORCE2 gene as drug target in preparation of drugs for treating Th17 cell-related diseases |
| CN113896757A (en) * | 2020-07-07 | 2022-01-07 | 中国科学院上海药物研究所 | Pentacyclic triterpene carbon glycoside compounds, and preparation method and application thereof |
| WO2022007765A1 (en) * | 2020-07-07 | 2022-01-13 | 中国科学院上海药物研究所 | Pentacyclic triterpenoid glycoside compound, and preparation method therefor and use thereof |
| CN113318230A (en) * | 2021-06-11 | 2021-08-31 | 上海交通大学医学院附属第九人民医院 | Application of Optineurin in diagnosis and treatment of ocular melanoma |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109481684B (en) | 2020-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109481684A (en) | Application using Optineurin as target spot in preparation prevention and treatment autoimmune disease drug | |
| Meli et al. | Role of innate immune response in non-alcoholic fatty liver disease: metabolic complications and therapeutic tools | |
| Luck et al. | Regulation of obesity-related insulin resistance with gut anti-inflammatory agents | |
| Smits et al. | Protective effect of Schistosoma mansoni infection on allergic airway inflammation depends on the intensity and chronicity of infection | |
| EP3165227B2 (en) | Composition for treating or preventing metabolic disease, containing, as active ingredient, extracellular vesicles derived from akkermansia muciniphila bacteria | |
| Kimura et al. | Protection of human corneal epithelial cells from TNF-α–induced disruption of barrier function by rebamipide | |
| Kumar et al. | Contagious pustular dermatitis (orf disease)–epidemiology, diagnosis, control and public health concerns | |
| CN107007626A (en) | Method and composition for cell therapy | |
| US10105329B2 (en) | Modulation of regulatory T cells via G-coupled protein receptor 43 | |
| Duan et al. | Antiviral effects of ergosterol peroxide in a pig model of porcine deltacoronavirus (PDCoV) infection involves modulation of apoptosis and tight junction in the small intestine | |
| Tian et al. | IL-17 down-regulates the immunosuppressive capacity of olfactory ecto-mesenchymal stem cells in murine collagen-induced arthritis | |
| Mercer et al. | Anorexia in rats infected with the nematode, Nippostrongylus brasiliensis: experimental manipulations | |
| Beer et al. | Evaluation of live-attenuated Histomonas meleagridis isolates as vaccine candidates against wild-type challenge | |
| Yang et al. | Deprivation of dietary fiber enhances susceptibility of piglets to lung immune stress | |
| CN110099688A (en) | Bone myohypertrophia inducer | |
| Wang et al. | Sishen wan treats ulcerative colitis in rats by regulating gut microbiota and restoring the Treg/Th17 balance | |
| Wang et al. | Role of TLR4/NF-κB pathway in the damage of acute hypobaric hypoxia to small intestinal mucosa in rats. | |
| Case et al. | Histamine H1 receptor signaling regulates effector T cell responses and susceptibility to coxsackievirus B3-induced myocarditis | |
| Zhu et al. | Artesunate interfere in modulation of Foxp3 expression in synovial cells in collagen-induced arthritis rats | |
| Enokida et al. | Oral administration of Lactiplantibacillus plantarum 22A-3 exerts anti-allergic activity against intestinal food allergy mouse models sensitized and challenged with ovalbumin | |
| CN107137711A (en) | Application of the albumen of Kindlin 2 as target spot in the medicine for treating Osteoarthritis is prepared | |
| EP3845242A1 (en) | Immune tolerance-inducing agent and therapeutic or prophylactic agent for allergic disorder | |
| Niizawa et al. | Clinical and immunomodulatory effects of fun-boi, an herbal medicine, on collagen-induced arthritis in vivo | |
| Liao et al. | Extracellular polysaccharides from Sporidiobolus pararoseus alleviates rheumatoid through ameliorating gut barrier function and gut microbiota | |
| US9526729B2 (en) | Medicament for treating peripheral neuropathies |
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 |