CN108815513A - Phosphoprotein phosphatase PPM1L is preparing the purposes in the drug for preventing or treating myocardial infarction - Google Patents

Abstract

The invention belongs to the field of biomedicine and provides the use of protein phosphatase PPM1L in the preparation of medicines for preventing or treating myocardial infarction. The present invention provides the use and strategy of this molecule in heart disease, especially for the prevention and treatment of myocardial infarction and heart failure caused by myocardial ischemia. The present invention proves that this molecule can inhibit the increase of inflammation caused by myocardial infarction and related factors both in vivo and in vitro, exert the anti-inflammatory effect in the early stage of myocardial infarction, improve the heart function and the survival rate of the body after myocardial infarction, and protect the heart after myocardial infarction. Protection from inflammatory damage.

Description

Protein phosphatase PPM1L in the preparation of medicines for preventing or treating myocardial infarction the use of

technical field

The invention belongs to the field of biomedicine, and relates to a protein phosphatase PPM1L, specifically a use of the protein phosphatase PPM1L.

Background technique

Myocardial infarction (MI) is acute coronary occlusion, which can lead to ischemic necrosis of myocardial cells [AT Turer. et al., Am J Cardiol. 2010; 106:360-368.]. Necrotic cardiomyocytes will trigger an early inflammatory response, and the responding cells include leukocytes and endothelial cells, which further recruit inflammatory cells and regulate inflammatory responses by expressing adhesion molecules, chemokines, and inflammatory cytokines [LiaudetL. et al., Front Biosci ( Schol Ed). 2013; 5:86-104.]. This inflammatory response is triggered by the activation of the innate immune response by endogenous substances released by necrotic cardiomyocytes. It is an effective adaptive response for the heart to respond to acute injury, and it is important for the repair of damage after myocardial infarction and the reconstruction of myocardial function. Significance [Liaudet L. et al., Front Biosci (Schol Ed). 2013; 5:86-104. Arslan F. et al., Nat Rev Cardiol. 2011; 8:292-300. SFrantz. et al., Cardiovasc Res. 2009; 81:474 -481.].

The intensity of the inflammatory response after myocardial infarction has a key influence on the outcome of myocardial infarction [Liaudet L. et al., Front Biosci (Schol Ed). 2013; 5:86-104. Arslan F. et al., Nat Rev Cardiol.2011; 8: 292-300.]. Too strong or too weak inflammatory response has adverse effects on cardiac remodeling after myocardial infarction. Excessive inflammatory response will aggravate the degradation of extracellular matrix, leading to heart rupture; continuous inflammatory response will reduce collagen deposition, resulting in weakened tension strength of scar formation, thereby increasing ventricular dilation; overexpressed inflammatory mediators will also activate apoptosis signaling pathways, Lead to myocardial cell apoptosis and reduce the number; on the contrary, too weak inflammatory response will cause inflammation to spread to non-infarcted myocardial tissue, resulting in myocardial fibrosis and weakened myocardial diastolic function [Frangogiannis NG.Circ Res.2012; 110:159-173 .]. In short, the dysregulation of inflammatory response after myocardial infarction can lead to persistent increase of myocardial infarct size, myocardial cell fibrosis, reactive myocardial hypertrophy and ventricular dilation, collectively referred to as adverse cardiac remodeling (Adverse cardiac remodeling), the outcome is functional heart failure ^{[ 2,5]} [Liaudet L. et al., Front Biosci (Schol Ed). 2013; 5:86-104. Frangogiannis NG. Circ Res. 2012; 110:159-173.]. Therefore, the inflammatory response after myocardial infarction is strictly regulated, however, the molecular regulation mechanism is still not very clear.

Activation of the innate immune system is the main reason for triggering the inflammatory response after myocardial infarction. Endogenous danger signals will be quickly captured by the natural immune response system in the first place, and then activate non-specific inflammatory responses. The activation of the natural immune system has a keen "receptor", called pattern recognition receptors (Pattern recognition receptors, PRRs), including Toll-like receptors (Toll-like receptors, TLRs) and NOD-like receptors (NOD-like receptors) , it is they that are responsible for identifying danger signals and activating downstream inflammatory signaling pathways [LA O'Neill. et al., NatRev Immunol.2007; 7:353-364. K Schroder. et al., Cell.2010; 140:821-832.].

Necrotic cardiomyocytes and extracellular matrix produced by myocardial infarction will release a variety of endogenous danger signal molecules, including HMGB1 (The Protein High Mobility Group Box-1) [M Andrassy. et al., Circulation.2008; 117:3216-3226 .], Heat Shock Proteins (HSPs)【Y Li. et al., J Biol Chem.2011; 286:31308-31319.N Zou. et al., Am J Physiol Heart CircPhysiol.2008; 294:H2805-2813. ] and the cell's own nucleic acid, ribonucleoprotein [F Arslan. et al., Nat RevCardiol.2011; These endogenous risk molecules are also called damage-associated molecular patterns (DAMPs), which can be recognized by pattern recognition receptors expressed by immune cells and endothelial cells ^[11] [F Arslan. et al., Nat RevCardiol.2011 8:292-300.], and then activate intracellular NF-κB, MAPKs signaling pathway and formation of inflammatory complex (inflammasome), thereby activating the expression of inflammatory cytokines, chemokines, growth factors and adhesion molecules, and recruiting Inflammatory cells go to the injury site, remove damaged tissue and dead cells, promote angiogenesis and fibroblast proliferation, and ultimately promote scar formation and cardiac remodeling. As the damaged tissue and dead cells are gradually phagocytized and cleared, some inflammatory inhibitory mediators are released, leading to the termination of the inflammatory response [Liaudet L. et al., Front Biosci (Schol Ed). 2013; 5:86-104. Arslan F. et al., Nat Rev Cardiol. 2011; 8:292-300. R Gill. et al., Free Radic Biol Med. 2010; 48:1121-1132.].

The TLR signaling pathway is strictly regulated, and various regulatory mechanisms are involved in the regulation of the innate immune response triggered by TLR [Xingguang Liu, et al., Nat Immunol; 2011; 12(5):416-24.J Immunol; 2010; 185:7244- 7251]. Among them, the phosphorylation process of signal protein is very important in the TLR signal transduction pathway, because it can regulate the activity and function of signal protein, and phosphatase is the enzyme that controls the phosphorylation level of signal protein in these signal transduction pathways. Essential [O'Neill LA. Immunity. 2008; 29:12-20.]. It has been reported in the literature that tyrosine phosphatase SHP-1 (Srchomology phosphatase-1) can inhibit the production of inflammatory cytokines triggered by TLR by directly regulating the phosphorylation levels of NF-κB and MAPKs, and can also inhibit the production of inflammatory cytokines by inhibiting IRAK-1. (Interleukin-1receptor associatedkinase-1) activation to inhibit the production of inflammatory cytokines triggered by TLR [An H, Hou J. et al., NatImmunol.2008; 9:542-550.]; SHP-2 (SH2-containing protein Tyrosine phosphatase2) can inhibit the production of cytokines by inhibiting the activation of TBK1 (TANK-binding kinase 1) in the TRIF pathway [AnH. et al., Immunity.2006; 25:919-928.].

PPM1L, also known as PP2Cε, contains PP2Cc (Serine/threonine phosphatases, family 2C, catalytic domain) functional domain, which is a serine/threonine phosphatase belonging to the PP2C subfamily. Mouse PPM1L encodes 360 amino acids and is widely expressed in mouse tissues [Li MG. et al., J Biol Chem. 2003; 278:12013-12021.]. So far, little is known about the regulatory role of PPM1L in inflammation, and its regulatory function in post-MI inflammation remains unreported.

Contents of the invention

Aiming at the above-mentioned technical problems in the prior art, the present invention provides the use of a protein phosphatase PPM1L, the use of the protein phosphatase PPM1L is to solve the ineffectiveness of drugs in the prior art for preventing and treating myocardial infarction. Good technical question.

The invention provides the use of protein phosphatase PPM1L in the preparation of medicaments for preventing or treating myocardial infarction.

Further, the manifestation factors of the myocardial infarction are excessive production of inflammatory factors after myocardial infarction, ventricular wall rupture or heart failure.

Further, the myocardial infarction is caused by coronary artery occlusion.

Further, the inflammatory factor is selected from TNFα, IL-1, IL-6 or IL-12.

Further, the protein phosphatase PPM1L is selected from

(a) the amino acid sequence of SEQ ID NO: 2;

(b) homologous to the amino acid sequence of SEQ ID NO: 2, which has a protein that inhibits damage-associated pattern molecule (DAMP)-induced related diseases and/or symptoms;

(c) Substitution, deletion or addition of one or several amino acids in the amino acid sequence of (a) or (b), and the activity of preventing or treating myocardial infarction-related diseases and/or symptoms derived from (a) or (b) of proteins or peptides.

Further, the coding gene of the protein phosphatase PPM1L is selected from:

(i) the sequence of SEQ ID NO: 1; or

(ii) a molecule that hybridizes to the sequence defined in (i) under stringent conditions and has the ability to prevent or treat myocardial infarction-related diseases and/or symptoms.

The present invention also provides a pharmaceutical composition comprising:

(A) a therapeutically effective amount of protein phosphatase PPM1L or the coding sequence; and

(B) A pharmaceutically or immunologically acceptable carrier or excipient.

Further, the protein phosphatase PPM1L and its coding sequence in the pharmaceutical composition account for 0.001-99.9 wt% of the total weight of the pharmaceutical composition.

Further, the protein phosphatase PPM1L and its coding sequence in the pharmaceutical composition account for 1-95 wt%, preferably 5-90 wt%, more preferably 10-80 wt%, of the total weight of the pharmaceutical composition.

The invention provides a new application of the protein phosphatase PPM1L protein or its coding sequence in effectively resisting myocardial infarction and inhibiting the production of inflammatory factors.

The expression of the protein phosphatase PPM1L of the present invention in myocardial tissue in the early myocardial infarction border zone is significantly reduced; in addition, the protein phosphatase PPM1L of the present invention can inhibit the early inflammatory response of myocardial infarction, thereby exerting anti-inflammatory, improving cardiac function and reducing The role of myocardial injury. The protein phosphatase PPM1L of the invention can be used to prepare medicines for inhibiting the early inflammation of myocardial infarction and the development of the disease course.

We studied the regulatory effect of PPM1L on the inflammatory response after myocardial infarction in the mouse model of myocardial infarction. The experimental results showed that the expression of PPM1L mRNA in the ischemic/infarcted area of the myocardial tissue was higher than that in the myocardial infarction model constructed by ligation of the left coronary artery of the mouse heart. Non-ischemic/infarcted tissue and myocardial tissue of sham-operated control mice significantly decreased; at the same time, overexpression of PPM1L significantly inhibited the expression of inflammatory factors in myocardial tissue induced by myocardial infarction; The production of inflammatory cytokines induced by HMGB1 in vivo was significantly lower than that in control mice; the cardiac function of PPM1L transgenic mice after myocardial infarction was significantly improved compared with wild-type mice, and the necrosis of myocardial ischemic injury was significantly reduced.

Through a large number of studies and animal model experiments, the inventors found that PPM1L can effectively inhibit the production of inflammatory factors after myocardial infarction, improve cardiac function and increase survival rate in myocardial infarction. The present inventors have completed the present invention on this basis.

Specifically, applied research on inflammation-related genes is a hotspot in the research of molecular biology and cell biology of inflammation. Applying nucleotides and proteins of inflammation-suppressing genes to the prevention and treatment of inflammation is an effective technique for artificially intervening in myocardial infarction. Therefore, it has broad application prospects in both functional genomics research and inflammation-related gene therapy.

The inventors have found through research that stable overexpression of PPM1L can reduce the production of inflammatory factors. In the mouse myocardial infarction model induced by ligation of the descending artery, it was observed that overexpressed PPM1L could significantly reduce the myocardial infarct size after ligation and the expression of inflammatory factors in serum and tissue, reduce myocardial injury, and protect heart function, suggesting that PPM1L may have The application prospect of treating myocardial infarction and other diseases. Thus, the present invention provides methods and strategies for applying PPM1L molecules to the prevention and treatment of myocardial infarction

According to the anti-inflammatory molecule PPM1L with anti-inflammatory effect, the present invention studies the production of inflammatory factors in macrophages in immune cells under the action of endogenous risk factors, and verifies that the sequence of the molecule can be used to treat myocardial infarction animals. Healing and protective effects. The experiments proved that 1) the expression of PPM1L in myocardial tissue was significantly reduced after myocardial infarction in mice; 2) PPM1L overexpressed in PPM1L transgenic mice significantly inhibited the production of tissue inflammatory factors; 3) in peritoneal macrophages derived from PPM1L transgenic mice The overexpression of PPM1L significantly inhibited the production of inflammatory factors induced by DAMPs; 4) The cardiac function of PPM1L transgenic mice after myocardial infarction was significantly improved compared with that of wild-type mice, and the necrosis of myocardial ischemic injury was significantly reduced.

Compared with the prior art, the technical progress of the present invention is remarkable. The anti-inflammatory molecular protein phosphatase PPM1L of the present invention can be applied to the prevention and treatment of myocardial infarction and heart failure caused by myocardial ischemia. The present invention proves that protein phosphatase PPM1L can inhibit the increase of inflammation caused by myocardial infarction and related factors both in vivo and in vitro, play an anti-inflammatory role in the early stage of myocardial infarction, protect heart function, improve the survival rate of the body after myocardial infarction, and protect the heart in the myocardium. Protection from inflammatory injury after infarction.

Description of drawings

Figure 1: The expression of PPM1L in myocardial tissue after myocardial infarction in mice (results are shown as mean ± standard deviation, n≥3; *, p<0.05; **, p<0.01)

Figure 2: Expression of inflammatory factor mRNA levels in myocardial tissue of PPM1L transgenic mice and wild-type mice after myocardial infarction (results are shown as mean ± standard deviation, n≥3; *, p<0.05; **, p< 0.01; ***, p<0.001)

Figure 3: DAMPs-induced changes in the protein levels of macrophage inflammatory factors from PPM1L transgenic mice and wild-type mice (results are shown as mean ± standard deviation, n≥3; *, p<0.05; **, p <0.01; ***, p<0.001)

Figure 4: PPM1L transgenic mice and wild-type mice after myocardial infarction changes in cardiac function and infarct size and comparison (results are shown as mean ± standard deviation, n≥3; *, p<0.05; **, p<0.01 ; ***, p<0.001)

Detailed ways

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods not indicating specific conditions in the following examples are usually according to conventional conditions such as the conditions described in Sambrook et al. "Molecular Cloning: Laboratory Guide" (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer suggested conditions. Percentages and parts are by weight unless otherwise indicated.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. In addition, any methods and materials similar or equivalent to those described can also be applied in the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

Example 1: Down-regulation of PPM1L expression in myocardial tissue after myocardial infarction in mice

Mice (C57 strain, male, 8-10 weeks old, purchased from Shanghai Xipuer-Bikay Experimental Animal Co., Ltd.) established myocardial infarction models according to the methods reported in the literature [Tarnavski, O. et al., Physiol Genomics.2004; 16; 349- 360], the mice in the sham operation group were only threaded without ligation of the anterior descending artery when the myocardial infarction model was established. Myocardial tissue from the border area of myocardial infarction and the normal area of the mice were collected at 1 day, 3 days, 5 days, 7 days, and 14 days after myocardial infarction, respectively.

The expression of PPM1L in myocardial tissue in the border zone of myocardial infarction was analyzed by real-time quantitative reverse transcription-PCR (qRT-PCR). Tissue total RNA was extracted using TRIzol (Invitrogen). qRT-PCR was performed on an ABI7900 (Applied Biosystems) real-time quantitative PCR instrument using SYBR Green RT-PCR kit (TOYOBO).

PPM1L quantitative PCR primers are: 5'-CGCGGATCCGATGATAGAGGATACAATGACTTTGC-3'(upstream)5'-CGGGGTACCGAGTGCTCTTCTGTTTTGCTACTA-3'(downstream)

The internal reference GAPDH quantitative PCR primers were 5'-AGGTCGGTGTGAACGGATTTG-3' (upstream) and 5'-TGTAGACCATGTAGTTGAGGTCA-3' (downstream).

The relative quantification of mRNA was calculated using the 2-ΔΔCt method (GAPDH was used as an internal reference) [Livak, KJ. et al., Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method.Methods.2001; 25:402-408] .

The expression of PPM1L in the myocardial tissue of the myocardial infarction border zone and normal zone after infarction in mice is shown in Figure 1. The results showed that the expression of PPM1L in the myocardial tissue of the myocardial infarction border area and normal area of the mice after infarction was significantly reduced at 1 day, 3 days, and 5 days, and gradually recovered to the initial level over time.

Example 2: Overexpression of PPM1L inhibits the production of inflammatory cytokines in myocardial tissue in the marginal zone of myocardial infarction in mice

Construct PPM1L transgenic mice (commissioned by East China Normal University) to overexpress the PPM1L gene. The myocardial infarction model was established in PPM1L transgenic mice, and the mice in the sham operation group were only threaded without ligation of the anterior descending branch when establishing the myocardial infarction model. Myocardial tissues from the border area of myocardial infarction and the normal area of the mice were collected 1 day and 3 days after myocardial infarction, respectively.

The expression of PPM1L in myocardial tissue in the border zone of myocardial infarction was analyzed by real-time quantitative reverse transcription-PCR (qRT-PCR). Tissue total RNA was extracted using TRIzol (Invitrogen). qRT-PCR was performed on an ABI7900 (Applied Biosystems) real-time quantitative PCR instrument using SYBR Green RT-PCR kit (TOYOBO).

IL-1β quantitative PCR primers are: 5'-GGTGTGTGACGTTCCCATTAGAC-3' (upstream) 5'-CATGGAGAATATCACTTGTTGGTTGA-3' (downstream)

IL-6 quantitative PCR primers are: 5'-TAGTCCTTCCTACCCCAATTTCC-3' (upstream) 5'-TTGGTCCTTAGCCACTCCTTC-3' (downstream)

TNF-α quantitative PCR primers are: 5'-AAGCCTGTAGCCCACGTCGTA-3' (upstream) 5'-GGCACCACTAGTTGGTTGTCTTTG-3' (downstream)

IL-12b quantitative PCR primers are: 5'-ACCCTGACCATCCAAGTCAAA-3' (upstream) 5'-TTGGCCTCGCATCTTAGAAAG-3' (downstream)

The results are shown in Figure 2: the levels of inflammatory factors in the peripheral zone myocardial tissue of PPM1L transgenic mice with myocardial infarction were significantly lower than those of wild-type mice. The results indicated that PPM1L transgene can inhibit the production of inflammatory factors in myocardial tissue after myocardial infarction.

Example 3: PPM1L overexpressed in peritoneal macrophages derived from PPM1L transgenic mice significantly inhibits the production of inflammatory factors induced by DAMPs

Wild-type and PPM1L transgenic mice were injected intraperitoneally with 2 mL broth (purchased from sigma), and 2 days later, the peritoneal cavity was washed with serum-free RPMI1640 (purchased from Corning Company) to obtain macrophages. Then the cells were stimulated with 0.1 mg/ml recombinant mouse HMGB1 or HSP60 for 0, 3, and 6 hours, and the cell supernatant was collected for ELISA (purchased from R&D Company) analysis.

The test results of the effect of PPM1L transgene on the production of inflammatory factors in mouse peritoneal macrophages are shown in Figure 3. The results showed that the protein levels of inflammatory factors expressed in peritoneal macrophages of PPM1L transgenic mice were significantly decreased after being stimulated by DAMPs. The results indicated that the PPM1L transgene could inhibit the production of inflammatory factors in mouse-derived peritoneal macrophages.

Example 4: Overexpression of PPM1L can significantly improve cardiac function and reduce myocardial infarction size in mice after myocardial infarction

The myocardial infarction model was established in PPM1L transgenic mice, and the mice in the sham operation group were only threaded without ligation of the anterior descending branch when establishing the myocardial infarction model.

The heart function of the mice was detected by small animal ultrasound (Vevo 2100, VISUALSONICS) at 3 days, 7 days, and 14 days after myocardial infarction. In addition, transgenic and wild-type hearts were harvested one week after myocardial infarction, and TTC staining was performed to detect the size of myocardial infarction.

Cardiac function and TTC staining results are shown in Figures 4A and 4B: after myocardial infarction, the cardiac function of PPM1L transgenic mice was significantly better than that of wild-type mice, and the area of myocardial infarction in PPM1L transgenic mice was significantly smaller than that of wild-type mice. The results showed that overexpression of PPM1L could significantly improve cardiac function after myocardial infarction and reduce myocardial infarction size.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

sequence listing

<110> Shanghai Oriental Hospital

<120> Use of protein phosphatase PPM1L in the preparation of drugs for preventing or treating myocardial infarction

<141> 2018-09-19

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aactagactg ttcataccct aatagttccc caaaattgcc ttagcatgtc acaccagcat 4980

ttgtcatcca gctctggaga aatgacatga ctgttagata ctatgcctgt ttaattgcct 5040

ctggattaag tcattgatag ctagactttt gagctagtta gctgtagaaa taatagaatc 5100

cagtgtttcc caaagtgtgttccaaagaac acaagttcca aagatgctct tctaaatata 5160

ggattctagg atgaaataag tttggaaaac acataccatc ttagaaagtt agtgaacatg 5220

agcacatcaa aggctctaat ttaaatgtcc tgtgaggaga gaaaaacaca acaaatatct 5280

actgtcagtt agcccagtgt tttccagatt aatttgacca cagaaacttt tttgagtggc 5340

acctagtaac actgcaagga acttgtgttc taaagaacac cctttgggaa attctggtat 5400

attgaaatat tgctgtgttt tcattcacga tgactcttag tagcagtaca atttgcaact 5460

tagaagcatg agcctttcat atatgaagct gacttgttaa taaaagcagt gttaagaagt 5520

agtatgactt aatatgacca acagcagcct catattgata gcagaacaat cctacttaaa 5580

gctctaaaca tcatcccccc cttttttttt ttaacggaat ctcgctctgt cacccaggct 5640

ggaatgccgt ggcgcaatgt caggtcactg caagctccgc ctcccaggtt cacaccattc 5700

tcctgcctca gcctcctgag cagctgggac tacaggcggc ctccaccaca cctggctaat 5760

tttttgtatt tttagtagag acggggtttc accatgttag ccaggatggt cttgatatcc 5820

tgaccttgta atccaccagc cttggcctcc caaagtgctg ggattacagg tgtgagccac 5880

tgcacccggc catccctttc ttttatatga ataatgagca agagccctgc catagctaaa 5940

tatgtctcaa attcattcca agatctttcc attgcttttt tgcagttacc tttgcttttt 6000

gggttgaatc aaaatgaata tttttatatt tcattactat atgatattta aaaatataca 6060

gaaaatcaca gaaaaggaaa aacagaaatt tgattttaat tcacttttga aattttaacg 6120

atttttaaaa ctaatgatct gttatataat aactgaaatg taaactatta acagttattt 6180

actttctttc ctttattgtc ctctctggat tcagtgaaat aatttgaaat caattagagc 6240

tcataccttc taaaactcct gtcccatatc atcccacctt tcatgtttta tgtacatagt 6300

taagtttggc atgttatctc ttgcctaaaa tgtgaggcct tcctgtagtt ccacagtagt 6360

tactcttggc tgcagaaata ggtttgggaa gctatgagag attaccccaa agtcatggat 6420

gaaacaaatc tagtcgaaaa catggtacag agtgaattaa ggcaaaagtc attaacttaa 6480

aattacatcc aatatttagg aatagctcat cctctgcaca ttatccaaaa tattttaaaa 6540

aactaaaagt aagatctctt aggctaggtg gcggattgca aagggcagggg ggcagttgat 6600

accatgaagc ttccaataga tgcatttcta aacatttttt ctaatatgta aatttgctga 6660

tccttagtaa tgttaatgct ttgtctattt gttattttta ccactgttga acagcaaact 6720

gttgaggcta gggttgatta ttgataattt ttatgtgttt agtaataacc acagcatcat 6780

acgtacaact gctttaataa atccaagtta aggactttga ggagagcatg tagcaagtat 6840

tcattttaac aaatcatcac agacagtttc cctgttcaaa gctccaccaa aaagcaagca 6900

tgcagcaaag gcatagactg tatttagaga tgtggtttca tttattcttg aagtcctata 6960

accttgcatt atttaaacac aaaatcccac ctaacatttg ccaaatccaa gttaatctca 7020

aaaacctccg aggacattga gcacaagcag attackgtaa gggcactggt ggcagattgt 7080

ggtggagaga gggctgagtg accccagagca aagcctgccc gggttactcg ctgccacttc 7140

aggagaggga ttgtatcagc tctcattacc ttttgtcatg aggcagttca taaattaaat 7200

atgatatcat gcattatctc aaaatatttc tatattaatt agataattca cgatgtaaaa 7260

cgtgttcatg cagaagcaaa ggcggatgta atagtcaagt atcagggaaa aaccccttta 7320

ggatacccag acattaaaaa tgtaaaaagt agcttataag aaatgtggat tatcagattt 7380

ttactatata taggttgtgt ttaatagtaa tttctgacac ctgcacatag atgaagaaaa 7440

ggcagttgtt accacttttc accattcaaa aaatggggctt gtctcttaga gtattggtta 7500

atgctttgct tatttgttaa tgaagacagc attttaattt taaagcttcc ttttgtagga 7560

ctgatggcac gggcagaatc acatttaaga aaaatggttt ggaacacttt ttaataaaaa 7620

attgtaaaat cagtttccag tgtggtcaaa attagaatgt aaaggaaagc atttttaaga 7680

aaatacagaa gcccaaatca agggagagta atataaaagg aaatatttta aattaagaaa 7740

aggaggtgtt atagtttatt acatcatgaa ttatgctttc tgtacttacg ctcttagtga 7800

aggagataac attgaagtga gaaaatgaac attctcagtt atttgcgaag ttagtaaaat 7860

tgcagtccct attccatttt tttctgatga aaatggatac atgagatctc atttattgct 7920

ctcctcaaca cctttactgt cccagattcc attcacgttc caaatcgtgg acagtaaaac 7980

tgaagtcaga taaacacagt cacaggtaca actgggagcg agttttaaca tagtggtaat 8040

tttgcaaatg ccaacatgaa tactagtgag tactcaattt acgacagaca atttcacaaa 8100

aaccaaccac agagtgattt gatattcttg cttcttagag agttttctaa tccatataat 8160

gcaatgggtc aggttgctca tatgattaaa aaaattaatc ctgtagaaat gatgtattag 8220

acatctctaa cccaagaaaa agatgtcttt tgtataactt ttaaattccc atttgtctat 8280

cttaggacaa atgatgactt cttcgtgtag taatgaaaac gtctaaatcc tgctctgtgg 8340

agaaaagcta gaaccatggg acattaagca agaatactcc tctgatttag gctcccagac 8400

tacaactaac agtctttatg tattatatcat cttggttat gttttatctc aagcatatac 8460

agtgtgcaga aagtagaata atgaataaaa tatgaccatg gagaaaaccc ataacttttg 8520

cttctattat agaaagcagt gtttgttctt tttaatttat tacatgttca cattgaaaca 8580

ttaacatcta caatattgtc tttcagttta ctcttcaatt actaaagcaa atgattttga 8640

tttatagcac ttacacaact gcagttcagg tcacagaatt gaaatctatt tgataatgtc 8700

tcagcattct catgggcagc caggaaaaaa aaaaagagag agaaagacaa atagaaattt 8760

tagtgggttc agataaatga ttattctgac aaacctggaa aatagtaatt aaacagttca 8820

ttccaaaaga ccccaatgta aactctgctc acatacataa tttgtataag gaaaatgtat 8880

gagttttttt cttaaaaata aatagaagca tcattgcatt tataacagat tgatcaatct 8940

tatgaacaga ttacattggg aaagtcacta ttattgctgt gtgtttgtat attttattct 9000

ttcctttttt gtgtgagcat ctactatgtg ccaggcagtg actgtgtctg agtgaatgta 9060

tccatggtga gcactcaggc gccatcactg ccttcttggg gcttacaccc tattggggtg 9120

tcagagatat tataacactc aatattgacc caatgggatg aaaattgtta tcttcatgat 9180

ttcttccaat gggcttagat ttgttgggac agggcagaat tagaataatt atttctcaca 9240

tataagagta agtgatatga gattgggaat ttcttgccaa cactcaagca aactgcagaa 9300

gataattact ccattttact agctcaccta gtttctgaag gatggggtgg ggagtgagac 9360

atggagcctc tggggacaag gatagggggg tgggactgtc ccagtggagg gtgccactgg 9420

aagcctccac ccaagtgggc cagcagctgt gcaagggtgc agcagtaggc tggtgaaagc 9480

caggttgtcc agatctttct aggggaagag aaggattata gaaaaggaga aaagggagga 9540

gcaagaacat ctctccatgg cctattttta tttttgttcc caagagacta gtgtaataac 9600

cttatgcccc caaagacagc ccagaactga gtatcttcag tgttcctgag tgtaaatcag 9660

tgaatctgtt atatagaaag ttaacatttc aggagcacaa ttaatttcctg atttcaaata 9720

tgaacagaag actaaatgtc atttttttaa atcataaatt tggtatccat agttacttta 9780

cacattccag agtctaagca tggaatttaa gttttgtatt ttgaacaaaa atagcaacat 9840

ctgttttctg tatggcaaag aaaaactaaa tcccagactt ttatttttaa ccacagaaac 9900

catcacttct ttctaaatta tttgcttgcc tgctcatttc ccctaaacat ctctgtttat 9960

gccaatggct ttttaaatcc aaataatact agttatgcc aaattgcatt agaatcctat 10020

tgaattagaa taattcatat acttcatata ctaggcagta aatggttaga gacggtaaag 10080

atctgattgt gatcaaaagt agggtttttg ttttgttttt aatcatgagc acatataaaa 10140

ggaaattcaa atgaaaccaa gccaatttct gatttagtaa gattaaaacc atagatggag 10200

ctcagttatt aaggatggaa aggaggaaag cacagagtga ggagagaaaa aggggaagga 10260

aaaaaagtag tgtttttttg gggaacagta taatgtctca gaagttcttc tctcttatcc 10320

tgctgatgtt gagacagtgg ctttgtataa gcttttattt atacctattt caacatcatg 10380

aaaatttttc catcagcatt ttcttgcaac tttaaaaatc acaagggatt atttgtttat 10440

aaagagatac aggatctata atttattggc aatactttat tttaggaatc ttcaaattgt 10500

acttaataaa taaatcacag gtcaaaatga ctgtaaaatc aatcaggctt tctcgtaagt 10560

tatttcctga aaaccatcca tgataaatac aacaaggctt ggcgaaacct cattactctt 10620

ggaagtcttc agaggttatt atacctactg atctgtattt gtctctaaga gacatttgaa 10680

acacagttac aatgtcaatt tggtgtaaca ttacctgtag tagagctcag agtatatggc 10740

agtgcacggt cactcaagat gcatggcgtg tcatttgggt cgctttgatg ggtgctcggg 10800

tagaattagc cttgtagaga ctaatgtgtt catgctatct ctgatcctgt ggtgttgtat 10860

aaaaatgaac agctaaaaat ttacccatga caagattaaa gcaaaaataa acacaaagtt 10920

gttagacttt aaacatgaaaaaaaa 10945

<210> 2

<211> 360

<212> PRT

<213> Homo sapiens

<400> 2

Met Ile Glu Asp Thr Met Thr Leu Leu Ser Leu Leu Gly Arg Ile Met

1 5 10 15

Arg Tyr Phe Leu Leu Arg Pro Glu Thr Leu Phe Leu Leu Cys Ile Ser

20 25 30

Leu Ala Leu Trp Ser Tyr Phe Phe His Thr Asp Glu Val Lys Thr Ile

35 40 45

Val Lys Ser Ser Arg Asp Ala Val Lys Met Val Lys Gly Lys Val Ala

50 55 60

Glu Ile Met Gln Asn Asp Arg Leu Gly Gly Leu Asp Val Leu Glu Ala

65 70 75 80

Glu Phe Ser Lys Thr Trp Glu Phe Lys Asn His Asn Val Ala Val Tyr

85 90 95

Ser Ile Gln Gly Arg Arg Asp His Met Glu Asp Arg Phe Glu Val Leu

100 105 110

Thr Asp Leu Ala Asn Lys Thr His Pro Ser Ile Phe Gly Ile Phe Asp

115 120 125

Gly His Gly Gly Glu Thr Ala Ala Glu Tyr Val Lys Ser Arg Leu Pro

130 135 140

Glu Ala Leu Lys Gln His Leu Gln Asp Tyr Glu Lys Asp Lys Glu Asn

145 150 155 160

Ser Val Leu Ser Tyr Gln Thr Ile Leu Glu Gln Gln Ile Leu Ser Ile

165 170 175

Asp Arg Glu Met Leu Glu Lys Leu Thr Val Ser Tyr Asp Glu Ala Gly

180 185 190

Thr Thr Cys Leu Ile Ala Leu Leu Ser Asp Lys Asp Leu Thr Val Ala

195 200 205

Asn Val Gly Asp Ser Arg Gly Val Leu Cys Asp Lys Asp Gly Asn Ala

210 215 220

Ile Pro Leu Ser His Asp His Lys Pro Tyr Gln Leu Lys Glu Arg Lys

225 230 235 240

Arg Ile Lys Arg Ala Gly Gly Phe Ile Ser Phe Asn Gly Ser Trp Arg

245 250 255

Val Gln Gly Ile Leu Ala Met Ser Arg Ser Leu Gly Asp Tyr Pro Leu

260 265 270

Lys Asn Leu Asn Val Val Ile Pro Asp Pro Asp Ile Leu Thr Phe Asp

275 280 285

Leu Asp Lys Leu Gln Pro Glu Phe Met Ile Leu Ala Ser Asp Gly Leu

290 295 300

Trp Asp Ala Phe Ser Asn Glu Glu Ala Val Arg Phe Ile Lys Glu Arg

305 310 315 320

Leu Asp Glu Pro His Phe Gly Ala Lys Ser Ile Val Leu Gln Ser Phe

325 330 335

Tyr Arg Gly Cys Pro Asp Asn Ile Thr Val Met Val Val Lys Phe Arg

340 345 350

Asn Ser Ser Lys Thr Glu Glu Gln

355 360

<210> 3

<211> 35

<212>DNA

<213> Mus musculus

<400> 3

cgcggatccg atgatagagg atacaatgac tttgc 35

<210> 4

<211> 33

<212>DNA

<213> Mus musculus

<400> 4

cggggtaccg agtgctcttc tgttttgcta cta 33

<210> 5

<211> 21

<212>DNA

<213> Mus musculus

<400> 5

aggtcggtgt gaacggattt g 21

<210> 7

<211> 23

<212>DNA

<213> Mus musculus

<400> 7

tgtagaccat gtagttgagg tca 23

<210> 6

<211> 23

<212>DNA

<213> Mus musculus

<400> 6

ggtgtgtgac gttcccatta gac 23

<210> 8

<211> 26

<212>DNA

<213> Mus musculus

<400> 8

catggagaat atcacttgtt ggttga 26

<210> 9

<211> 23

<212>DNA

<213> Mus musculus

<400> 9

tagtccttcc taccccaatt tcc 23

<210> 10

<211> 21

<212>DNA

<213> Mus musculus

<400> 10

ttggtcctta gccactccctt c 21

<210> 11

<211> 21

<212>DNA

<213> Mus musculus

<400> 11

aagcctgtag cccacgtcgt a 21

<210> 12

<211> 24

<212>DNA

<213> Mus musculus

<400> 12

ggcaccacta gttggttgtc tttg 24

<210> 13

<211> 21

<212>DNA

<213> Mus musculus

<400> 13

accctgacca tccaagtcaa a 21

<210> 14

<211> 21

<212>DNA

<213> Mus musculus

<400> 14

ttggcctcgc atcttagaaa g 21

Claims (11)

1. phosphoprotein phosphatase PPM1L is preparing the purposes in the drug for preventing or treating myocardial infarction.

2. purposes as described in claim 1, which is characterized in that the performance factor of the myocardial infarction is inflammation after myocardial infarction Excess generation, ventricle wall rupture or the cardiac failure of the factor.

3. purposes as described in claim 1, which is characterized in that the myocardial infarction is caused by coronary occlusion ischemic 's.

4. purposes as claimed in claim 2, which is characterized in that the inflammatory factor be selected from TNF α,

IL-1, IL-6 or IL-12.

5. purposes as described in claim 1, which is characterized in that the phosphoprotein phosphatase PPM1L is selected from

(a)SEQ ID NO：2 amino acid sequence；

(b) with SEQ ID NO：2 amino acid sequence homologous have and damage associative mode molecule (DAMP) are inhibited to induce related disease Disease and/or the active albumen of symptom；

(c) in (a) or amino acid sequence (b) through replacing, missing or adding one or several amino acid and have prevention or Treat myocardial infarction related disease and/or symptom it is active as (a) or (b) derived from protein or polypeptide.

6. purposes as described in claim 1, which is characterized in that the encoding gene of the phosphoprotein phosphatase PPM1L is selected from:

(i)SEQ ID NO：1 sequence；Or

(ii) under strict conditions with (i) limit sequence hybridize and have prevention or treat myocardial infarction related disease and/or The molecule of symptom.

7. a kind of pharmaceutical composition, it includes：

(A) the phosphoprotein phosphatase PPM1L or coded sequence of therapeutically effective amount；And

(B) acceptable carrier or excipient pharmaceutically or in immunology.

8. pharmaceutical composition as claimed in claim 7, which is characterized in that phosphoprotein phosphatase PPM1L in described pharmaceutical composition And its coded sequence accounts for 0.001~99.9wt% of pharmaceutical composition total weight.

9. pharmaceutical composition as claimed in claim 8, which is characterized in that phosphoprotein phosphatase PPM1L in described pharmaceutical composition And its coded sequence accounts for 1~95wt% of pharmaceutical composition total weight, preferably 5~90wt%, more preferable 10~80wt%.

10. a kind of any pharmaceutical composition inhibits myocardial infarction Earlier period of inflammation in preparation or delays in claim 7-9 Application in the drug of myocardial infarction disease.

11. phosphoprotein phosphatase PPM1L inhibits myocardial infarction Earlier period of inflammation in preparation or delays the drug of myocardial infarction disease In application.