CN109646668B - A kind of polypeptide is used to prepare the purposes of prevention and treatment Alzheimer disease drugs - Google Patents
A kind of polypeptide is used to prepare the purposes of prevention and treatment Alzheimer disease drugs Download PDFInfo
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- CN109646668B CN109646668B CN201910007568.9A CN201910007568A CN109646668B CN 109646668 B CN109646668 B CN 109646668B CN 201910007568 A CN201910007568 A CN 201910007568A CN 109646668 B CN109646668 B CN 109646668B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
- A61K38/1774—Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
Abstract
The invention discloses the purposes that a kind of polypeptide is used to prepare prevention and treatment Alzheimer disease drugs.Its amino acid sequence is as shown in SEQ ID NO:1, and the polypeptide is the extracellular fragment 41-81 amino acids of TREM2 receptor protein, and nucleotide sequence is as shown in SEQ ID NO:2.The polypeptide has the inflammatory reaction for promoting small colloid to mediate, the apoptosis for inhibiting microglia enhances the transfer ability of microglia, promotes endocytosis and degradation of the microglia to A β, the density for increasing big intracerebral microglia reduces the purposes of big intracerebral A beta-amyloyd spot deposition.
Description
Technical field
The present invention relates to fields of biomedicine more particularly to a kind of polypeptide to be used to prepare prevention and treatment Alzheimer disease drugs
Purposes.
Background technique
Alzheimer disease (Alzheimer ' s Disease, AD) is that a kind of lost with gradual cognitive function is main
The neurodegenerative disease of feature is most commonly seen senile dementia type.AD is after cardiovascular disease, cerebrovascular disease and cancer
Afterwards, " the 4th killer " of the elder's health.With population in the world aging, AD becoming 21st century maximum disease it
One.However, U.S. FDA and European Union EMA only ratify 5 kinds of AD drugs so far, own although having paid huge scientific effort
These drugs are all symptomatic treatment, and the progress of the AD state of an illness can be prevented or delayed without one kind.Therefore, new AD medicine is found
Object target spot simultaneously develops targeted drug and has extremely important economy and society meaning.
There is the poor loss of memory, confusion of consciousness, attention, aphasia, appraxia, the symptoms such as sense of lacking direction early stage in AD patient,
Gradually lose thinking and judgement, it is difficult to link up with people, life can not take care of oneself.Finally because nerve cell mortality influences
When most of brain area, body respectively organizes impaired organ, and patient occurs dead.Patient's AD postmortem is found in its brain there are two tools
Main pathological characteristics: neurofibrillary tangles (Neurofibrillary tangles, NFTs) and nerve cell in neuron
Outer amyloid plaque (Amyloid plaques) deposition.Mainly by beta amyloid peptide, (Amyloid- β, is commonly abbreviated as amyloid plaque
A β) assemble, A β, which is deposited, causes the degeneration and Neuron Apoptosis of nerve synapse in brain, it is final cause cerebral damage and
It is dull-witted.Thus, inhibit A β to generate or A β degradation is promoted to have great importance the prevention and treatment of AD.In addition to above two main disease
It manages outside feature, there are also other pathological characters for the big intracerebral of patient AD: including neuroinflamation excessive activation, neuron loss, nerve
Synaptic loss etc..
2 type myeloid cells triggering receptor (Triggering receptor expressed on myeloid cells 2,
TREM2) belong to Ig (Immunoglobulin, Ig) superfamily, encoding gene is located on chromosome 6p21, the nucleotide of coding
It is NM_018965.3 with number of the amino acid sequence in Genbank.TREM2 is newfound AD risk genes in recent years,
Coding region mutation R47H can make the onset risk of late hair style AD increase by 3 times or so, the No.1 risks and assumptions with AD --- APOE4's
Risk is suitable.TREM2 is mainly expressed on the microglia in brain, and microglia is extremely important congenital
Immunity-associated cell, providing first layer for central nervous system is also most important one layer of defence line, resists outer infections, is maintained
Brain stable state.In AD, microglia participates in and adjusts Neuroinflammation, monitors ambient enviroment in real time, migrates and assemble
To lesions position, the A β deposition and impaired neuron removed in central nervous system delay process so as to improve AD symptom.
The microglia function that more and more evidences support TREM2 to mediate, which maintains, plays highly important protective effect in AD,
Thus TREM2 becomes the new popular target of AD medicament research and development.
Summary of the invention
The purpose of the present invention is to provide a kind of polypeptides with prevention and treatment Alzheimer disease.
To achieve the above object, the present invention provides a kind of amino acid sequence polypeptide as shown in SEQ ID NO:1 for making
The purposes of standby prevention and treatment Alzheimer disease drugs.
Further, the polypeptide is the extracellular fragment 41-81 amino acids of TREM2 receptor protein, nucleotide sequence such as SEQ
Shown in ID NO:2.
Further, the polypeptide has the purposes for increasing microglia proinflammatory reaction.
Further, the polypeptide has the purposes for inhibiting microglia apoptosis.
Further, the polypeptide has the purposes for increasing microglia migration.
Further, the polypeptide, which has, increases microglia to the purposes of A β endocytosis.
Further, the polypeptide, which has, increases the purposes that microglia degrades to A β.
Further, the polypeptide has the purposes for increasing brain No microglial density.
Further, the polypeptide has the purposes for reducing A beta-amyloyd spot number in brain.
On the other hand, the present invention also provides a kind of this recombinant vectors of the polypeptide to be used to prepare prevention and treatment Alzheimer disease
The purposes of drug.
The amino acid sequence of TREM2 41-81 are as follows:
MKHWGRRKAWCRQLGEKGPCQRVVSTHNLWLLSFLRRWNGS SEQ ID NO:1；
The corresponding nucleotide sequence of TREM2 41-81 are as follows:
ATGAAGCACTGGGGGAGGCGCAAGGCCTGGTGCCGCCAGCTGGGAGAGAAGGGCCCATGCCAGCGTGT
GGTCAGCACGCACAACTTGTGGCTGCTGTCCTTCCTGAGGAGGTGGAATGGGAGC SEQ ID NO:2.
It can be obtained by embodiment, TREM2 41-81 polypeptide can significantly improve the multiple functions of microglia, embody
: promote inflammatory reaction, inhibit Apoptosis, promotes cell migration, and increase microglia to the endocytosis of oligomer A β 42
And degradation.STREM2 41-81 polypeptide is injected into APP/PS1 mouse brain by Naoliqing capsule, can dramatically increase hippocampus
The density of region microglia, and significantly reduce the amyloid plaque deposition of hippocampus.
Detailed description of the invention
Fig. 1 is the restriction enzyme mapping of pFUSE-hIgG1-Fc1 carrier；
Fig. 2 is the SDS-PAGE silver staining electrophoretogram of sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein；
Fig. 3 is that sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein promotes microglia inflammatory reaction
Result figure；
Fig. 4 is the result that sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein inhibits microglia apoptosis
Figure；
Fig. 5 is the result that sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein promotes microglia migration
Figure；
Fig. 6 is that sTREM2 41-81-Fc fusion protein increases microglia to the endocytosis figure of A β 42；
Fig. 7 is that sTREM2 41-81-Fc fusion protein increases microglia to the degradation figure of A β 42；
Fig. 8 is the result figure that sTREM2 41-81-Fc fusion protein increases hippocampus microglia density；
Fig. 9 is the result figure that sTREM2 41-81-Fc fusion protein reduces hippocampus A beta-amyloyd spot deposition.
Specific embodiment
The embodiment of the present invention is described below in detail, examples of the embodiments are shown in the accompanying drawings, wherein from beginning to end
Same or similar label indicates same or similar element or element with the same or similar functions.Below with reference to attached
The embodiment of figure description is exemplary, it is intended to is used to explain the present invention, and is not considered as limiting the invention.Embodiment
In particular technique or condition person is not specified, described technology or conditions or according to the description of product according to the literature in the art
Book carries out.Reagents or instruments used without specified manufacturer, being can be with conventional products that are commercially available.
The preparation of embodiment 1:sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein
The building of pFUSE-sTREM2 1-171-hIgG1-Fc1 expression plasmid: selecting pFUSE-hIgG1-Fc1, (purchase is certainly
Invivogen, article No. pfuse-hg1fc1, carrier picture are shown in Fig. 1) it is carrier for expression of eukaryon, cloning site selects I He of EcoR
I restriction enzyme site of Xho constructs pFUSE-sTREM2 1-171-hIgG1-Fc1 expression plasmid.Steps are as follows: design upstream primer:
5’-CGGAATTCATGGAGCCTCTCCGGCTGC-3 ' SEQ ID NO:3, wherein underscore is I restriction enzyme site of EcoR；Draw in downstream
Object: 5 '-CCGCTCGAGTTTGGGAAGGGGATTTCTC-3 ' SEQ ID NO:4, wherein underscore is I restriction enzyme site of Xho.
The nucleotides sequence of TREM2 be classified as (purchase from Sino Biological Inc., article No. HG11084-M:
TREM-2 cDNA ORF Clone in Cloning Vector, Human):
Wherein single underscore is the signal peptide sequence of albumen, and wave underline is sTREM2 41-81 amino acid, double lower strokes
Line is the cross-film (TM) and intracellular section of corresponding nucleotide sequence of albumen.
With cDNA (SEQ ID NO:5) be template, can be artificial synthesized, SEQ ID NO:3 be upstream primer, SEQ ID NO:
4 be downstream primer, and PCR clones sTREM2 1-171 expressing gene, glue recovery purifying purpose product.Recycle restriction enzyme
I digestion pFUSE-hIgG1-Fc1 plasmid of EcoR I and Xho, glue recycle large fragment, carry out digestion simultaneously to PCR product with identical enzyme
Glue recovery purifying digestion products connect sTREM2 1-171 digestion products and pFUSE-hIgG1-Fc1 enzyme using T4DNA ligase
Product is cut, obtains recombinant plasmid pFUSE-sTREM2 1-171-hIgG1-Fc1, conversion bacillus coli DH 5 alpha competent cell is simultaneously
Culture selects positive colony, and send raw work bioengineering (Shanghai) share limited public affairs in the culture medium containing Zeocin resistance
Department's sequencing confirmation encoder block is correct, obtains correct recombinant plasmid and is named as pFUSE-sTREM21-171-hIgG1-Fc1.
The building of pFUSE-sTREM2 41-81-hIgG1-Fc1 expression plasmid: selection pFUSE-hIgG1-Fc1 is eukaryon table
Up to carrier, cloning site selects EcoR I and I restriction enzyme site of Xho, and building pFUSE-sTREM2 41-81-hIgG1-Fc1 expresses matter
Grain.Steps are as follows: design upstream primer: 5'-CAGAGCTGTCCGGAGCCATGAAGCACTGGGGGAGGCG-3’SEQ ID
NO:6, wherein underscore is part signal peptide sequence；Downstream primer: 5 '-CCGCTCGAGTTGCTCCCATTCCACCTCCTCAG
SEQ ID NO:7, wherein underscore is I restriction enzyme site of Xho；Artificial synthesized signal peptide Sense sequences: 5'-ATGGAGCCTCTCC
GGCTGCTCATCTTACTCTTTGTCACAGAGCTGTCCGGAGCC-3 ' SEQ ID NO:8, artificial synthesized signal peptide antisense sequence
Arrange 5'-GGCTCCGGACAGCTCTGTGACAAAGAGTAAGATGAGCAGCCGGAGAGGCTCCA T-3 ' SEQ ID NO:9.
With cDNA (SEQ ID NO:5) be template, can be artificial synthesized, SEQ ID NO:6 be upstream primer, SEQ ID NO:
7 be sTREM2 41-81 expressing gene of the downstream primer PCR clone comprising part signal peptide sequence, and glue recovery purifying purpose produces
Object.By artificial synthesized SEQ ID NO:8 and SEQ ID NO:9 mixed in equal amounts, 5min is reacted under the conditions of 95 DEG C, is slowly dropped to
The signal peptide complete sequence double-strand of room temperature acquisition renaturation.With the sTREM2 41-81 expression comprising part signal peptide sequence of purifying
The signal peptide complete sequence double-strand of gene and renaturation is template, and SEQ ID NO:3 is upstream primer, and SEQ ID NO:7 is downstream
Primer PCR clone includes the sTREM2 41-81 expressing gene of signal peptide sequence, glue recovery purifying purpose product.Recycle limitation
Property restriction endonuclease EcoR I and I digestion pFUSE-hIgG1-Fc1 plasmid of Xho, glue recycle large fragment, with identical enzyme to PCR product into
Row digestion and glue recovery purifying digestion products connect sTREM2 41-81 digestion products and pFUSE- using T4DNA ligase
HIgG1-Fc1 digestion products obtain recombinant plasmid pFUSE-sTREM2 41-81-hIgG1-Fc1, convert bacillus coli DH 5 alpha sense
It by state cell and cultivates in the culture medium containing Zeocin resistance, selects positive colony, and send raw work bioengineering (Shanghai)
Limited liability company's sequencing confirmation encoder block is correct, obtains correct recombinant plasmid and is named as pFUSE-sTREM241-81-
hIgG1-Fc1。
The expression and purifying of sTREM2 1-171-Fc and sTREM2 41-81-Fc fusion protein: by identification correctly recombination
Plasmid is purified, using TurboFect transfection reagent by this plasmid transfection into HEK 293T cell line.It, will after 24 hours
The DMEM that culture medium is replaced with serum-free continues culture 24 hours, and collection culture medium is simultaneously broken by 0.45 μm of filter membrane removal cell
Culture medium is incubated overnight by piece with 4 DEG C of Protein A- pearl, and 1000g is centrifuged 5 minutes in 4 DEG C, and precipitating is taken to wash using PBS
Protein A- pearl is three times.Add IgG Elution Buffer (purchase from Thermo Fisher) vibrate 10 seconds after,
10000g is centrifuged 30 seconds, supernatant is added contains 1M pH8.0Tris-HCl solution (the IgG Elution of 1/10 volume rapidly
Buffer it is neutralized in centrifuge tube), 2 μ L drops is taken to be verified after mixing in pH test paper.Protein solution is placed in 10kDa's
It dialyses in dialysis membrane and in 4 DEG C of PBS solutions.Using 10kDa protein concentration pipe (Millipore, UFC801024) from
Heart protein concentrate, resulting sTREM2 albumen measure protein concentration through SDS-PAGE electrophoresis, then carry out silver staining and identify that its is pure
Degree.For qualification result as shown in Fig. 2, fusion protein band is clear, purity is higher.
Embodiment 2:sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein is to microglia inflammatory reaction
Influence
With 5 × 105By primary microglia, (3-4 days C57BL/6N newborn mice brains divide cell number after birth
From obtaining) it is laid on 12 orifice plates and is cultivated.After 24 hours, respectively with the sTREM2 41-81-Fc fusion protein of 20nM,
(i.e. pFUSE-hIgG1-Fc1 is carried for sTREM21-171-Fc fusion protein (i.e. the resulting albumen of embodiment 1) and Fc reference protein
Body) it handles cell and continues culture 4 hours in serum free medium.Cell is placed on ice immediately and is washed with 1 × PBS is pre-chilled
3 times, total serum IgE is extracted using TRIzol reagent (Thermo Fisher, article No. 15596018), and use Reverse Transcriptase kit (north
Capital Quan Shijin, article No. AT314-02) reverse transcription is at cDNA.IL-6, TNF- are detected by Real-time quantitative PCR (RT-qPCR)
The mRNA level in-site of α, IL-1 β and β-Actin analyze other three kinds of inflammatory factors by internal reference of the mRNA level in-site of β-Actin
It is horizontal.As a result as shown in figure 3, after the processing of sTREM2 41-81-Fc fusion protein, IL-6, TNF-α, IL-1 β in microglia
Expression be respectively Fc processing 198.1,13.7 and 8.5 times of control group；And sTREM2 1-171-Fc fusion protein is handled
Afterwards, IL-6 in microglia, TNF-α, IL-1 β expression be respectively 35.8,5.8 and 4.1 times of Fc processing control group.
The above results explanation, sTREM2 41-81-Fc fusion protein can dramatically increase the inflammatory reaction of microglia, and it promotees
Scorching function is better than sTREM2 1-171-Fc fusion protein.
Influence of embodiment 3sTREM2 41-81-Fc and sTREM2 the 1-171-Fc fusion protein to microglia apoptosis
With 105Primary microglia is laid on the slide in 24 orifice plates (in advance with Poly-Lysine packet by cell number
By 24 hours) on cultivated.After 48 hours, sTREM2 41-80-Fc fusion protein, the sTREM2 1- of 20nM are used respectively
171-Fc fusion protein (i.e. 1 gained albumen of embodiment) and Fc reference protein (i.e. pFUSE-hIgG1-Fc1 carrier) handle cell
And continue culture 24 hours in serum free medium.Cell is placed on ice and is washed 3 times with 1 × PBS is pre-chilled, 4% poly first
After the fixed 20min of aldehyde room temperature, with 0.2%Triton X-100 permeable membrane 5min.It is situated between followed by terminal deoxynucleotidyl transferase
The dUTP method led carries out nick end label (TUNEL) detection kit (Promega, G3250) measurement apoptotic cell, is used in combination
Core dyestuff DAPI carries out core dye to analyze total cell number.Fluorescence microscope take pictures and count TUNEL positive cell number with
And DAPI positive cell number, the influence by comparing the ratio of the two with each albumen of quantitative analysis to microglia apoptosis.
As a result as shown in figure 4, the apoptosis ratio of Fc processing control group No microglial is 8.2%；And sTREM2 41-81-Fc and
After sTREM2 1-171-Fc fusion protein is handled respectively, the apoptosis ratio of microglia drops to 2.9% and 2.3% respectively,
Illustrate that sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein can significantly inhibit the apoptosis of microglia.
The shadow that 4 sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein of embodiment migrates microglia
It rings
With 105Primary microglia is laid on the cell Transwell (8 μm, Costar, article No. 3422) simultaneously by cell number
Culture 1 hour in serum free medium (100 μ L).Then in cell, lower layer is separately added into 600 μ L and contains 100nM sTREM2
41-80-Fc fusion protein, sTREM2 1-171-Fc fusion protein (i.e. the resulting albumen of embodiment 1) and Fc reference protein is (i.e.
PFUSE-hIgG1-Fc1 carrier) serum free medium, continue culture 24 hours after, cell is placed in and sucks culture medium on ice,
And washed 3 times with 1 × PBS of pre-cooling, after the fixed 20min of 4% paraformaldehyde room temperature, utilize HE staining kit (Beijing Suo Laibao, goods
Number G1120) it is dyed.Scrape off cell upper cell gently with cotton swab to detect the cell number migrated to lower membrane.In order to
Influence of the albumen to total cell number is excluded, (cotton is not used in the small ventricular cell in upper layer to the total cell number of statistics cell the upper and lower
Label wipe), then statistically analyze lower layer's migrating cell and total cell number purpose ratio.As a result as shown in figure 5, sTREM2 41-
After the processing of 81-Fc and sTREM2 1-171-Fc fusion protein, the mobility of microglia is respectively Fc processing control group
2.6 and 2.8 times, illustrate that sTREM2 41-81-Fc and sTREM2 1-171-Fc fusion protein can significantly increase microglia
Transfer ability.
Influence of the 5 sTREM2 41-81-Fc fusion protein of embodiment to microglia endocytosis and degradation A β 42
With the sTREM2 41-81-Fc fusion protein (i.e. the resulting albumen of embodiment 1) of 20nM and Fc reference protein is (i.e.
PFUSE-hIgG1-Fc1 carrier) the primary microglia of processing, the oligomeric forms with FAM fluorescence labels are added after 12 hours
A β 42 continue culture 3 hours after, collect cell and carry out flow cytomery fluorescence intensity intracellular.As a result as shown in fig. 6,
After the processing of sTREM2 41-81-Fc fusion protein, microglia is in the A β 42 of the oligomeric forms with FAM fluorescence labels
Horizontal 1.6 times for Fc control group are gulped down, illustrate that sTREM2 41-81-Fc fusion protein can dramatically increase microglia to widow
The endocytosis of aggressiveness A β 42.
With the sTREM2 41-81-Fc fusion protein (i.e. the resulting albumen of embodiment 1) of 20nM or Fc reference protein is (i.e.
PFUSE-hIgG1-Fc1 carrier) the primary microglia of processing, after culture 12 hours, 30 points are handled using lysosomal inhibitor
Clock, the A β 42 for adding oligomeric forms continue culture 3 hours, collect cell and crack, are detected and passed through using A β 42ELISA method
Cross the level of the microglia A β 42 intracellular of processing.The amount that A β 42 degrades is to be subtracted not pressing down by lysosomal inhibitor processing group
The level of preparation group obtains.As a result as shown in fig. 7, after the processing of sTREM2 41-81-Fc fusion protein, A β 42 in microglia
Degradation Level be 1.6 times of Fc control group, illustrate that sTREM2 41-81-Fc fusion protein albumen can dramatically increase small colloid
Degradation of the cell to oligomer A β 42.
6 sTREM2 41-81-Fc fusion protein of embodiment is to microglia density in APP/PS1 mouse brain and A β
The influence of amyloid plaque deposition
79 months big APP/PS1 mouse (4 heros 3 are female) are taken, isoflurane anesthesia is placed on stereotaxic apparatus, carefully
It cuts off scalp and finds bregma position, find position (hippocampus: ± 2.0mm, -2.2mm) by origin of bregma, and in this position
Punching.Albumen is injected with 0.2 μ L/min speed, depth 2.0mm using micro syringe.Half brain of left and right injects Fc control egg respectively
White (i.e. pFUSE-hIgG1-Fc1 carrier) or sTREM2 41-81-Fc fusion protein (i.e. the resulting albumen of embodiment 1), albumen
Concentration is 2 μ g/ μ L, each to inject 3 μ L.Allowing syringe needle to stop 5min after the completion of injection spreads albumen sufficiently, slowly extracts syringe out,
It sews up a wound and coats erythromycin, mouse is put back into rearging cage raising.After injection 7 days, brain tissue is collected, microglia is utilized
Specific antibody Iba1 (1:200, Wako, 019-19741) carries out immunofluorescence dyeing, as a result as shown in figure 8, injection Fc control
The region No microglial area of albumen is 1.1%, and it is thin to inject small colloid in the region of sTREM2 41-81-Fc fusion protein
Born of the same parents' area is 3.1%.The above results explanation, after sTREM2 41-81-Fc fusion protein is injected into hippocampus, can significantly increase
Add the density of microglia.It is immune that amyloid plaque is carried out using A β antibody (1:400, MOAB2, Abcam, ab126649) simultaneously
Fluorescent staining statisticallys analyze the distribution of A beta-amyloyd spot.As a result as shown in figure 9, the region A beta-amyloyd of injection Fc reference protein
Spot deposition 1.9%, the region A beta-amyloyd spot deposition 1.5% of injection sTREM2 41-81-Fc fusion protein.The above result shows that
After sTREM2 41-81-Fc fusion protein is injected into hippocampus, the A beta-amyloyd spot deposition in the region can be significantly reduced.
In conclusion TREM2 41-81 polypeptide can significantly improve the multiple functions of microglia, it is embodied in:
Promote inflammatory reaction, inhibit Apoptosis, promotes cell migration, and increase microglia to the endocytosis and drop of oligomer A β 42
Solution.STREM2 41-81 polypeptide is injected into APP/PS1 mouse brain by Naoliqing capsule, can dramatically increase hippocampus
The density of microglia, and significantly reduce the amyloid plaque deposition of hippocampus.Thus, TREM2 41-81 polypeptide or expression
The mRNA liposome of the albumen, DNA plasmid carrier carry the viral vectors of its encoding gene and prevent and treat alzheimer ' in preparation
Silent medicine field will have broad application prospects.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art are not departing from the principle of the present invention and objective
In the case where can make changes, modifications, alterations, and variations to the above described embodiments within the scope of the invention.
SEQUENCE LISTING
<110>Xiamen University
<120>a kind of polypeptide is used to prepare the purposes of prevention and treatment Alzheimer disease drugs
<130> XMDXN-19002-CNI
<160> 9
<170> PatentIn version 3.5
<210> 1
<211> 41
<212> PRT
<213>artificial synthesized
<400> 1
Met Lys His Trp Gly Arg Arg Lys Ala Trp Cys Arg Gln Leu Gly Glu
Lys Gly Pro Cys Gln Arg Val Val Ser Thr His Asn Leu Trp Leu Leu
Ser Phe Leu Arg Arg Trp Asn Gly Ser
<210> 2
<211> 123
<212> DNA
<213>artificial synthesized
<400> 2
atgaagcact gggggaggcg caaggcctgg tgccgccagc tgggagagaa gggcccatgc 60
cagcgtgtgg tcagcacgca caacttgtgg ctgctgtcct tcctgaggag gtggaatggg 120
agc 123
<210> 3
<211> 27
<212> DNA
<213>artificial synthesized
<400> 3
cggaattcat ggagcctctc cggctgc 27
<210> 4
<211> 28
<212> DNA
<213>artificial synthesized
<400> 4
ccgctcgagt ttgggaaggg gatttctc 28
<210> 5
<211> 693
<212> DNA
<213>artificial synthesized
<400> 5
atggagcctc tccggctgct catcttactc tttgtcacag agctgtccgg agcccacaac 60
accacagtgt tccagggcgt ggcgggccag tccctgcagg tgtcttgccc ctatgactcc 120
atgaagcact gggggaggcg caaggcctgg tgccgccagc tgggagagaa gggcccatgc 180
cagcgtgtgg tcagcacgca caacttgtgg ctgctgtcct tcctgaggag gtggaatggg 240
agcacagcca tcacagacga taccctgggt ggcactctca ccattacgct gcggaatcta 300
caaccccatg atgcgggtct ctaccagtgc cagagcctcc atggcagtga ggctgacacc 360
ctcaggaagg tcctggtgga ggtgctggca gaccccctgg atcaccggga tgctggagat 420
ctctggttcc ccggggagtc tgagagcttc gaggatgccc atgtggagca cagcatctcc 480
aggagcctct tggaaggaga aatccccttc ccacccactt ccatccttct cctcctggcc 540
tgcatctttc tcatcaagat tctagcagcc agcgccctct gggctgcagc ctggcatgga 600
cagaagccag ggacacatcc acccagtgaa ctggactgtg gccatgaccc agggtatcag 660
ctccaaactc tgccagggct gagagacacg tga 693
<210> 6
<211> 37
<212> DNA
<213>artificial synthesized
<400> 6
cagagctgtc cggagccatg aagcactggg ggaggcg 37
<210> 7
<211> 32
<212> DNA
<213>artificial synthesized
<400> 7
ccgctcgagt tgctcccatt ccacctcctc ag 32
<210> 8
<211> 54
<212> DNA
<213>artificial synthesized
<400> 8
atggagcctc tccggctgct catcttactc tttgtcacag agctgtccgg agcc 54
<210> 9
<211> 54
<212> DNA
<213>artificial synthesized
<400> 9
ggctccggac agctctgtga caaagagtaa gatgagcagc cggagaggct ccat 54
Claims (2)
1. the use that a kind of amino acid sequence polypeptide as shown in SEQ ID NO:1 is used to prepare prevention and treatment Alzheimer disease drugs
On the way；The polypeptide is the extracellular fragment 41-81 amino acids of TREM2 receptor protein, nucleotide sequence such as SEQ ID NO:2 institute
Show；The polypeptide has for increasing microglia proinflammatory reaction, microglia apoptosis, increase microglia being inhibited to move
It moves, increase microglia to A β endocytosis, increase microglia to A β degradation, increase brain No microglial density, drop
The function of A beta-amyloyd spot number in low brain.
2. the purposes that the recombinant vector of polypeptide described in expression claim 1 is used to prepare prevention and treatment Alzheimer disease drugs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910007568.9A CN109646668B (en) | 2019-01-04 | 2019-01-04 | A kind of polypeptide is used to prepare the purposes of prevention and treatment Alzheimer disease drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910007568.9A CN109646668B (en) | 2019-01-04 | 2019-01-04 | A kind of polypeptide is used to prepare the purposes of prevention and treatment Alzheimer disease drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109646668A CN109646668A (en) | 2019-04-19 |
| CN109646668B true CN109646668B (en) | 2019-10-18 |
Family
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| CN111000985A (en) * | 2020-01-02 | 2020-04-14 | 中山大学孙逸仙纪念医院 | Medicine for reducing starch deposition plaques in brains of AD patients |
| CN113214097B (en) * | 2020-01-21 | 2022-08-30 | 厦门大学 | Compounds for the treatment of alzheimer's disease |
| CN111184742B (en) * | 2020-02-13 | 2022-06-24 | 中山大学附属第五医院 | Application of TREM-2+ T cells in preparation of drugs for treating tuberculosis |
| CN113621078B (en) * | 2021-09-06 | 2023-03-28 | 山东大学齐鲁医院 | Fusion polypeptide TAT-pY682 and application thereof in preparation of Alzheimer disease drugs |
| CN115109138B (en) * | 2022-06-10 | 2024-02-02 | 武汉大学 | sTREM2 polypeptide analogue and application thereof in treatment of Alzheimer's disease |
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