CN110294808A - A kind of polypeptide SR4 and its application with anti-inflammatory effect - Google Patents

A kind of polypeptide SR4 and its application with anti-inflammatory effect Download PDF

Info

Publication number
CN110294808A
CN110294808A CN201910496590.4A CN201910496590A CN110294808A CN 110294808 A CN110294808 A CN 110294808A CN 201910496590 A CN201910496590 A CN 201910496590A CN 110294808 A CN110294808 A CN 110294808A
Authority
CN
China
Prior art keywords
polypeptide
inflammatory
mouse
inflammatory cytokine
tnf
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
CN201910496590.4A
Other languages
Chinese (zh)
Other versions
CN110294808B (en
Inventor
焦新安
潘志明
熊丹
宋丽
耿士忠
陈祥
黄金林
殷月兰
王晶
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.)
Yangzhou University
Original Assignee
Yangzhou University
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 Yangzhou University filed Critical Yangzhou University
Priority to CN201910496590.4A priority Critical patent/CN110294808B/en
Publication of CN110294808A publication Critical patent/CN110294808A/en
Application granted granted Critical
Publication of CN110294808B publication Critical patent/CN110294808B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
    • C07K14/255—Salmonella (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The polypeptide SR4 and its application that the invention discloses a kind of with anti-inflammatory effect, the anti-inflammatory effect of polypeptide SR4 is identified using the method that real-time fluorescence quantitative PCR and elisa technique combine, and identify in mouse model its infected by influenza attacks malicious protective effect.Specifically include that identification SR4 polypeptide to the inhibiting effect of Turnover of Mouse Peritoneal Macrophages inflammatory cytokine IL-1 β and TNF-α; evaluate influence of the SR4 polypeptide to cell activity; it explores SR4 polypeptide and attacks malicious protective effect in the intracorporal anti-inflammatory effect of mouse and its infected by influenza.SR4 polypeptide can significantly inhibit the inflammatory cytokine IL-1 β of LPS induction and the generation of TNF-α; and maintain good cell bio-activity; the secretion of inflammatory cytokine IL-6, IL-12p40 and TNF-α that excess LPS can be inhibited to induce in Mice Body, and protect the attack of mouse resistance influenza virus.The polypeptide has the function of inhibiting inflammatory reaction and resists pathogen infection, this will provide new approaches for the design of novel suppression inflammation small-molecule drug.

Description

A kind of polypeptide SR4 and its application with anti-inflammatory effect
Technical field
The invention belongs to polypeptide functional study fields, and in particular to a kind of interference inflammatory signal access simultaneously inhibits inflammatory reaction Polypeptide function identification.
Background technique
Defence line is immunized as first of body in natural immune system, can pass through pattern recognition receptors (Pattern Recognition receptors, PRRs) identification pathogen-associated molecular pattern (Pathogen-associated molecular Patterns, PAMPs), expression of nuclear factor kappa B (Nuclear factor κ B, NF κ B) signal path is activated, induction generates anticusp Property cell factor and antibacterial peptide resist the invasion of pathogenic microorganism.Toll-like receptor (Toll-like receptors, TLRs) It is a kind of typical PRRs, PAMP-TLR interaction causes receptor by the change of TIR structural domain dimerization or occurred conformation, Then the TIR structural domain of receptor combines the adaptor protein of the structural domain containing TIR, such as MyD88 (Myeloid differentiation Factor 88) and TRIF (TIR domain-containing adaptor protein including interferon- β), start downstream signal Cascaded amplification, cause the transposition of Nuclear Factor kappa B, lead to pro-inflammatory cytokines, in active oxygen The up-regulation of mesosome and costimulatory molecules expression quantity.
Studies have shown that the polypeptide of TIR structural domain is capable of the adaptor protein of the competitive binding structural domain intracellular containing TIR, thus Normal TIR-TIR interaction is interfered, blocks TLRs-NF κ B access, and then inhibit the secretion of inflammatory cytokine and chemotactic factor (CF). 4R3 (TLR4-TIR region 3) polypeptide can significantly inhibit point of the Turnover of Mouse Peritoneal Macrophages inflammatory factor of LPS induction Secrete, the polypeptide of the TIR structural domain of TIRAP/Mal is able to suppress TLR2 and TLR4 signal path, and can reduce in Mice Body by Inflammatory reaction caused by LPS.The BB ring that MyD88TIR structural domain is such as simulated using polypeptide finds that this polypeptide can weaken mouse The generation and toxicity of the pro-inflammatory cytokines induced in vivo by staphylococcus enterotoxin B.Many infectious diseases itself are exempted from Epidemic disease and malignant tumour can all cause body and generate excessive inflammatory response, thus target the medicine of innate immunity and signal transmitting Object has potential medical prospect.In the microbial patients with sepsis of Gram-negative, TLR4 antagonist can be used for preventing because Endotoxic shock caused by LPS.TLRs antagonist can be used for treating autoimmune disease, especially systemic loupus erythematosus (Systemic lupus erythematosus,SLE).Therefore, the acology method for targeting TLRs may effectively inhibit The development and deterioration of autoimmune disease, identifying has potential medical prospect with the micromolecule polypeptide for pressing down scorching effect.
However, the receptor or adaptor protein that there is the polypeptide for pressing down scorching function to be all from eukaryocyte reported at present.Micro- life Object has evolved corresponding strategy to inhibit the inflammatory response of body, and the mechanism of bacterium escape TLRs-NF κ B signal access is The research of the following anti-inflammatory drugs provides theoretical basis, and the specific suppression inflammation polypeptide for screening bacterial origin will be inflammation related disease Treatment new drug candidate is provided.
Summary of the invention
In order to overcome the overactivity of TLRs signal path that body is caused to generate serious inflammatory reaction, the present invention provides one Kind has polypeptide SR4 and its application of anti-inflammatory effect.The present invention identifies one section of polypeptide SR4 with anti-inflammatory properties, and studies it Mouse is intracorporal anti-inflammatory and antiviral effect.
In order to achieve the above-mentioned object of the invention, the technical scheme adopted by the invention is as follows: a kind of polypeptide with anti-inflammatory effect The amino acid sequence of SR4, the polypeptide SR4 are RQIKIWFQNRRMKWKKVVLSKSFIKKDWTEYE (SEQ ID NO.1).
The present invention also provides SR4 polypeptide above-mentioned inhibit Turnover of Mouse Peritoneal Macrophages inflammatory cytokine production method, This method using Real-Time Fluorescent Quantitative PCR Technique identification SR4 polypeptide can inhibit LPS induce inflammatory cytokine IL-1 β and The generation of TNF-α, the specific steps are as follows:
(1) Salmonella enteritidis TcpS albumen TIR structural domain is divided by multiple sections, including sr4 by sequence alignment analysis； Introducing in the N section of polypeptide has the Antp homeodomain sequence RQIKIWFQNRRMKWKK of cell permeability to form polypeptide SR4；
(2) acquisition of Turnover of Mouse Peritoneal Macrophages；
(3) real-time fluorescence quantitative PCR identification SR4 polypeptide inhibits the generation of the inflammatory cytokine of LPS induction.
A further object of the present invention is to provide the pharmaceutical composition comprising polypeptide SR4 above-mentioned and preparing anti-inflammatory agent Purposes in object or anti-influenza virus medicament.
The present invention also provides polypeptide SR4 above-mentioned to prepare the purposes in anti-inflammatory drug or anti-influenza virus medicament.It is described The drug that anti-inflammatory drug specifically refers to interference TLR signal path and inflammatory cytokine is inhibited to secrete.
Compared with the existing technology, the TIR structural domain of TcpS albumen (NCBI sequence number: AIY53972.1) is divided by the present invention Multiple sections, by pressing down scorching effect and cell activity two in terms of carry out comprehensive selection；Current TcpS albumen passes through eukaryotic expression Plasmid-transfected cells are expressed, and prokaryotic expression system is with inclusion body expression (without soluble protein), and preparation process is more multiple It is miscellaneous.Relative to TcpS albumen, SR4 polypeptide of the invention is easily obtained (small peptide of 32 amino acid, chemical synthesis and quality control Technology maturation processed, purity is high, simple process is time saving, at low cost)；SR4 polypeptide action site is more single, and molecular weight is small, nonreactive Originality, few side effects are highly-safe；The N-terminal of SR4 polypeptide introduces the Antp homeodomain sequence with cell permeability, is easy to It absorbs, effect is efficient rapidly.
Detailed description of the invention
Fig. 1 is the different polypeptide sections of TcpS TIR structural domain.Wherein, sr1-sr6 is the difference of TcpS TIR structural domain Peptide fragment, cp are random polypeptide sequences.In addition, the N section of every segment polypeptide introduces the Antp homeodomain sequence with cell permeability RQIKIWFQNRRMKWKK is respectively formed SR1, SR2, SR3, SR4, SR5, SR6 and CP, to reinforce the ability that polypeptide enters cell.
Fig. 2 is the generation for the inflammatory cytokine that different SR polypeptides inhibit LPS to induce in vitro.Wherein, not homopolypeptide (40 μ M) effects Mice peritoneal macrophage respectively, after 30min, with 10ng/mL LPS stimulation, stimulates Trizol after 1h to crack, extracts RNA, qRT-PCR detect the expression of inflammatory cytokine IL-1 β (Fig. 2A) and TNF-α (Fig. 2 B), and wherein internal reference is β- actin。
Fig. 3 is that MTT experiment evaluates influence of each polypeptide to cell activity.Wherein, homopolypeptide (40 μM) is acted on respectively Turnover of Mouse Peritoneal Macrophages detects the bioactivity of cell by MTT experiment.
Fig. 4 is the secretion for inhibiting the inflammatory cytokine of LPS induction in SR4 polypeptide body.Wherein, SR4 polypeptide is injected intraperitoneally C57BL/6 mouse (10nmol/g), the LPS that sublethal dose is reused after 1h are attacked malicious (1 μ g/g), and ELISA detects blood after 4h In inflammatory cytokine IL-6 (Fig. 4 A), IL-12p40 (Fig. 4 B) and TNF-α (Fig. 4 C) secretion level.
Fig. 5 is the infection that SR4 polypeptide protects mouse resistance influenza virus.Wherein, with H1N1JS38 influenza virus collunarium sense C57BL/6 mouse is contaminated, SR4 polypeptide (10nmol/g) is injected intraperitoneally daily within the 2nd day to the 4th day after infection, daily to mouse Weight is monitored.
Specific embodiment
Further definition is of the invention in following embodiment, according to above description and these embodiments, those skilled in the art Member can determine essential characteristic of the invention, and without departing from the spirit and scope of the invention, can be to the present invention Various modifications and change are made, so that it uses various uses and condition.In addition to special indicate, of the present invention is this The field prior art.
Embodiment 1
SR4 polypeptide inhibits the generation of Turnover of Mouse Peritoneal Macrophages inflammatory cytokine, and this method uses real time fluorescent quantitative Round pcr identification SR4 polypeptide can significantly inhibit the generation of the inflammatory cytokine IL-1 β of LPS induction, the specific steps are as follows:
(1) Salmonella enteritidis TcpS albumen may interfere with TLRs signal path, inhibit the generation of inflammatory cytokine, to escape The natural immune system of ease host.TcpS albumen TIR structural domain is divided into multiple sections by sequence alignment analysis, respectively Sr1, sr2, sr3, sr4, sr5 and sr6, while rondom polypeptide cp is set as negative control (Fig. 1).In addition, to reinforce polypeptide Into the ability of cell, the Antp homeodomain sequence with cell permeability is introduced in the N section of every segment polypeptide RQIKIWFQNRRMKWKK is respectively formed SR1, SR2, SR3, SR4, SR5, SR6 and CP, and each Purity is >=95%.SR1's Amino acid sequence is RQIKIWFQNRRMKWKKTEDITESYDVFISH (SEQ ID NO.2), and the amino acid sequence of SR2 is RQIKIWFQNRRMKWKKAELLRAKGIN (SEQ ID NO.3), the amino acid sequence of SR3 is The amino acid sequence of RQIKIWFQNRRMKWKKVWYDEFSLGWGKSL (SEQ ID NO.4), SR4 are RQIKIWFQNRRMKWK The amino acid sequence of KVVLSKSFIKKDWTEYE (SEQ ID NO.1), SR5 are RQIKIWFQNRRMKWKKLKFSPTLVDKMAL (SEQ ID NO.5), the amino acid sequence of SR6 are RQIKIWFQNRRMKWKKIAEQLESLLK (SEQ ID NO.6), the ammonia of CP Base acid sequence is RQIKIWFQNRRMKWKKSLHGRGDPMEAFII (SEQ ID NO.7).
(2) acquisition of Turnover of Mouse Peritoneal Macrophages
By mouse euthanasia, 75% alcohol takes out mouse after impregnating 3min, is aseptically mentioned with the tweezers of sterilizing Mouse lower abdomen skin is played, an osculum is cut off with operating scissors, peritonaeum is completely exposed, is trained with syringe injection 10mL RPMI 1640 Base is supported, extracts peritoneal fluid back and forth, different directions rinse for several times, peritoneal fluid is finally sucked out, is transferred in 15mL centrifuge tube.Cell is outstanding Liquid 1000rpm is centrifuged 10min and has hanged cell with the RPMI 1640 containing 10%FBS after 1640 culture medium of RPMI washs one time, As Turnover of Mouse Peritoneal Macrophages.
(3) real-time fluorescence quantitative PCR identification SR4 polypeptide inhibits the generation of the inflammatory cytokine of LPS induction
By Turnover of Mouse Peritoneal Macrophages bed board (24 porocyte plates, 2 × 105A/hole), it is discarded after being incubated overnight not adherent Cell, is added Medium or not after (40 μM, such as SR1, SR2, SR3, SR4, SR5 and SR6) incubation 30min of homopolypeptide, with 10ng/mL LPS stimulation stimulates Trizol after 1h to crack, extracts RNA, and at cDNA, qRT-PCR detection is thin for reverse transcription after removing DNA The expression of intracellular cytokine IL-1 β and TNF-α, using β-actin as reference gene.QRT-PCR system is (20 μ L): FastStart Universal SYBR Green Master (Rox) 10 μ L, Forward primer (10 μM) 0.6 μ L, Reserve primer (10 μM) 0.6 μ L, 2 μ L of template (~30ng), RNase-free H2O 6.8μL.Reaction condition is (a) 95 DEG C, 30s；(b) 95 DEG C, 5s；(c) 60 DEG C, 34s, (b)-(c) 40 recycle.Primer sequence is β-actin, F: AGCCATGTACGTAGCCATCC, R:CTCTCAGCTGTGGTGGTGAA；IL-1 β, F:GCCCATCCTCTGTGACTCAT, R: AGGCCACAGGTATTTTGTCG；TNF-α, F:AGCCCCCAGTCTGTATCCTT, R:CTCCCTTTGCAGAACTCAGG；More than Primer sequence is respectively SEQ ID NO.8-13.LPS can induce the generation of a large amount of inflammatory cytokines as the result is shown, and polypeptide can Significantly inhibit the generation of the inflammatory cytokine of LPS induction, wherein the inhibiting rate to IL-1 β is respectively SR2 (64%), SR3 (84%), SR4 (73%), SR5 (72%), SR6 (84%), the inhibiting rate to TNF-α are respectively SR1 (63%), SR2 (90%), SR3 (93%), SR4 (96%), SR5 (94%), SR6 (95%) (Fig. 2).
Embodiment 2
SR4 polypeptide can maintain good cell activity: homopolypeptide being distinguished effects Mice peritoneal macrophage, is passed through The bioactivity of MTT experiment detection cell.Specific step is as follows:
By Turnover of Mouse Peritoneal Macrophages bed board (96 porocyte plates, 5 × 104A/hole), it is discarded after being incubated overnight not adherent Cell is added Medium or not after (40 μM) incubations 5h of homopolypeptide, 0.5mg/mL MTT reagent incubation 3h is added, discards culture Base is added 100 μ L Formazan solvents, 5min is leniently vibrated on shaking table, OD is read in microplate reader540It is each more to evaluate Influence of the peptide to cell activity.Polypeptide SR1, SR3 and SR4 can maintain preferable cell activity, cell activity as the result is shown Respectively SR1 (115%), SR3 (102%), SR4 (107%) (Fig. 3) make SR4 in conjunction with the suppression inflammation effect of above-mentioned each polypeptide It is candidate for optimal suppression inflammation polypeptide.
Embodiment 3
SR4 polypeptide can inhibit the secretion of mouse systemic inflammatory cytokine: by SR4 polypeptide intraperitoneal injection of mice, then infuse The LPS of sublethal dose is penetrated, the proinflammatory cytokines in mice serum are detected.Specific step is as follows:
C57BL/6 mouse (10nmol/g) is injected intraperitoneally in SR4 polypeptide, the LPS that sublethal dose is reused after 1h attacks poison (1 μ g/g), mice serum is taken after 4h, and ELISA detects point of inflammatory cytokine IL-6, IL-12p40 and TNF-α in serum Bleeding is flat, wherein IL-6 and IL-12p40ELISA kit is purchased from BD company (IL-6ELISA set:Cat#555240, IL- 12p40OPTEIA set:Cat#555165), TNF-α ELISA kit is purchased from BioLegend company (TNF-α ELISA MAXTM: Cat#430901).Specific steps are as follows: 1. overnight by 4 DEG C of each coated antibody coatings.2. secondary daily PBST adds after washing 5 times Enter confining liquid in 37 DEG C of closing 2h.3. PBST is washed 5 times, 100 μ L samples to be tested, 37 DEG C of incubation 2h are added in each hole.4. PBST is washed It washs 5 times, is separately added into corresponding detection antibody, 37 DEG C of incubation 1h.5. PBST is washed 5 times, 100 μ L streptavidin marks are added in each hole The horseradish peroxidase of note, 37 DEG C of incubation 30min.6. PBST is washed 5 times, the colour developing of 100 μ L tmb substrates 5min, 50 μ L is added 2M H2SO4Color development stopping, microplate reader read OD450Value.Inflammatory cytokine in a large amount of inducing mouse blood of LPS energy as the result is shown Secretion, and SR4 polypeptide can significantly inhibit LPS induction inflammatory cytokine generation, inhibiting rate is respectively IL-6 (78%), IL-12p40 (72%), TNF-α (58%), wherein IL-12p40 and TNF-α are even restored to normal level (Fig. 4).
Embodiment 4
SR4 polypeptide can protect the infection of mouse resistance influenza virus: after influenza infection mouse, by SR4 polypeptide abdominal cavity Inject mouse, the daily changes of weight for monitoring mouse.Specific step is as follows:
With 106EID50H1N1JS38 influenza virus collunarium infect C57BL/6 mouse, after infection the 2nd day to the 4th day daily It is injected intraperitoneally SR4 polypeptide (10nmol/g), the weight of mouse is monitored daily.The result shows that after influenza infection, The mouse weight of PBS control group and CP control group sharply declines that (the 10th day weight fall is respectively 27% He after infection 29%), and SR4 polypeptide can protect mouse to prevent its weight loss (the 10th day weight fall is only 5.5% after infection) (Fig. 5).
A kind of polypeptide SR4 with anti-inflammatory properties of the invention can significantly reduce the secretion of inflammatory cytokine, inhibit machine The inflammatory reaction of body, and preferable cell bio-activity is maintained, it is a kind of anti-inflammatory drug candidate of great potential, while being also new The design of type anti-inflammatory small-molecule drug provides new approaches.
Basic principles and main features and advantages of the present invention of the invention have been shown and described above.The skill of the industry Art personnel it should be appreciated that the present invention is not limited to the above embodiments, the above embodiments and description only describe The principle of the present invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these Changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and Its equivalent thereof.
Sequence table
<110>Yangzhou University
<120>a kind of polypeptide SR4 and its application with anti-inflammatory effect
<130> xhx2019061001
<141> 2019-06-10
<160> 13
<170> SIPOSequenceListing 1.0
<210> 1
<211> 32
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Val Val Leu Ser Lys Ser Phe Ile Lys Lys Asp Trp Thr Glu Tyr Glu
20 25 30
<210> 2
<211> 30
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Thr Glu Asp Ile Thr Glu Ser Tyr Asp Val Phe Ile Ser His
20 25 30
<210> 3
<211> 26
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Ala Glu Leu Leu Arg Ala Lys Gly Ile Asn
20 25
<210> 4
<211> 30
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Val Trp Tyr Asp Glu Phe Ser Leu Gly Trp Gly Lys Ser Leu
20 25 30
<210> 5
<211> 29
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Leu Lys Phe Ser Pro Thr Leu Val Asp Lys Met Ala Leu
20 25
<210> 6
<211> 26
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 6
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Ile Ala Glu Gln Leu Glu Ser Leu Leu Lys
20 25
<210> 7
<211> 30
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 7
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
Ser Leu His Gly Arg Gly Asp Pro Met Glu Ala Phe Ile Ile
20 25 30
<210> 8
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 8
agccatgtac gtagccatcc 20
<210> 9
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 9
ctctcagctg tggtggtgaa 20
<210> 10
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 10
gcccatcctc tgtgactcat 20
<210> 11
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 11
aggccacagg tattttgtcg 20
<210> 12
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 12
agcccccagt ctgtatcctt 20
<210> 13
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 13
ctccctttgc agaactcagg 20

Claims (6)

1. a kind of polypeptide SR4 with anti-inflammatory effect, which is characterized in that the amino acid sequence of the polypeptide SR4 is RQIKIW FQNRRMKWKKVVLSKSFIKKDWTEYE。
2. SR4 polypeptide described in claim 1 inhibits the production method of Turnover of Mouse Peritoneal Macrophages inflammatory cytokine, feature It is, the inflammatory cytokine IL-1 β that this method can inhibit LPS to induce using Real-Time Fluorescent Quantitative PCR Technique identification SR4 polypeptide With the generation of TNF-α, the specific steps are as follows:
(1) Salmonella enteritidis TcpS albumen TIR structural domain is divided by multiple sections, including sr4 by sequence alignment analysis；? The N section of sr4 polypeptide, which introduces, has the Antp homeodomain sequence RQIKIWFQNRRMKWKK of cell permeability to form polypeptide SR4；
(2) acquisition of Turnover of Mouse Peritoneal Macrophages；
(3) real-time fluorescence quantitative PCR identification SR4 polypeptide inhibits the generation of the inflammatory cytokine of LPS induction.
3. including the pharmaceutical composition of polypeptide SR4 described in claim 1.
4. polypeptide SR4 described in claim 1 is preparing the purposes in anti-inflammatory drug or anti-influenza virus medicament.
5. purposes according to claim 4, which is characterized in that the anti-inflammatory drug specifically refers to interference TLR signal path And the drug for inhibiting inflammatory cytokine to secrete.
6. pharmaceutical composition as claimed in claim 3 is preparing the purposes in anti-inflammatory drug or anti-influenza virus medicament.
CN201910496590.4A 2019-06-10 2019-06-10 Polypeptide SR4 with anti-inflammatory effect and application thereof Active CN110294808B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910496590.4A CN110294808B (en) 2019-06-10 2019-06-10 Polypeptide SR4 with anti-inflammatory effect and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910496590.4A CN110294808B (en) 2019-06-10 2019-06-10 Polypeptide SR4 with anti-inflammatory effect and application thereof

Publications (2)

Publication Number Publication Date
CN110294808A true CN110294808A (en) 2019-10-01
CN110294808B CN110294808B (en) 2022-11-18

Family

ID=68027795

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910496590.4A Active CN110294808B (en) 2019-06-10 2019-06-10 Polypeptide SR4 with anti-inflammatory effect and application thereof

Country Status (1)

Country Link
CN (1) CN110294808B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112898383A (en) * 2021-01-29 2021-06-04 深圳海创生物科技有限公司 Oligopeptide, active peptide composition and application of oligopeptide and active peptide composition in preparation of product with anti-inflammatory effect

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105985414A (en) * 2015-03-05 2016-10-05 扬州大学 Protein with anti-inflammatory effect and preparation and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105985414A (en) * 2015-03-05 2016-10-05 扬州大学 Protein with anti-inflammatory effect and preparation and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JUN ZOU等: "A TIR Domain Protein from E. faecalis Attenuates MyD88-Mediated Signaling and NF-kB Activation", 《PLOS ONE》 *
RUCHI M.NEWMAN等: "Identification and Characterization of a Novel Bacterial Virulence Factor That Shares Homology with Mammalian Toll/Interleukin-1 Receptor Family Proteins", 《INFECTION AND IMMUNITY》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112898383A (en) * 2021-01-29 2021-06-04 深圳海创生物科技有限公司 Oligopeptide, active peptide composition and application of oligopeptide and active peptide composition in preparation of product with anti-inflammatory effect

Also Published As

Publication number Publication date
CN110294808B (en) 2022-11-18

Similar Documents

Publication Publication Date Title
CN101155825B (en) Antimicrobial hexapeptides
CN106414480A (en) Polymyxin derivatives as antimicrobial compounds
Li et al. A teleost complement factor Ba possesses antimicrobial activity and inhibits bacterial infection in fish
Sun et al. The immune responses triggered by CpG ODNs in shrimp Litopenaeus vannamei are associated with LvTolls
Li et al. An enhanced variant designed from DLP4 cationic peptide against Staphylococcus aureus CVCC 546
CN110498848B (en) Melittin variant and application thereof
CN109833326A (en) Macrolide antibiotics is blocking the application in influenza infection
KR101749316B1 (en) A Highly Potent Broad-Spectrum Neutralizing Monoclonal Antibody Derived From H1N1-Infected Patients and Method Of Treatment of Virus By Using Thereof
CN110294808A (en) A kind of polypeptide SR4 and its application with anti-inflammatory effect
Luo et al. A short peptidoglycan recognition protein protects Boleophthalmus pectinirostris against bacterial infection via inhibiting bacterial activity
Yu et al. Effects of IBDV infection on expression of chTLRs in chicken bursa
CN102516394B (en) CMG 2mutant and Fc fusion protein, its encoding gene and its application
CN104628869A (en) Fusion peptide derivatives with antibacterial anti-influenza virus activities
CN107670023A (en) A kind of new application of V12CBD albumen and its encoding gene
Weerachatyanukul et al. Infectious hypodermal and hematopoietic necrosis virus-like particle (IHHNV-VLP) induces peroxiredoxin expression and activity in Fenneropenaeus merguiensis
CN101570760B (en) Recombinant mouse beta-alexin 3 polypeptide, preparation and use thereof
CN113754750A (en) Antibacterial peptide and application thereof in aquaculture
CN102115496A (en) Antimicrobial peptide Pc-CATH1 and gene thereof, chemical synthesis method and application thereof
CN115969976B (en) Application of FBXO6 protein or encoding gene thereof as target point
CN103965292B (en) Structure and application of Japanese encephalitis virus envelope protein binding peptide
TW201617088A (en) Antibacterial peptide and application thereof
CN116763902A (en) Use of anti-coronavirus lipopeptides for the treatment and prevention of influenza
CN105254736A (en) Cathelicidin family broad spectrum antimicrobial peptide Pb-CATH4 from python molurus bivittatus and gene, preparation and application thereof
Zhu et al. Antimicrobial and anti‐inflammatory activities of the neuropeptide antagonist SPA
CN116983391A (en) Treatment method for enhancing immunity and preventing and treating infectious diseases and complications thereof

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