CN104491879B - A kind of GEM 132 medical composition and its uses of miRNA 2861 - Google Patents
A kind of GEM 132 medical composition and its uses of miRNA 2861 Download PDFInfo
- Publication number
- CN104491879B CN104491879B CN201410803184.5A CN201410803184A CN104491879B CN 104491879 B CN104491879 B CN 104491879B CN 201410803184 A CN201410803184 A CN 201410803184A CN 104491879 B CN104491879 B CN 104491879B
- Authority
- CN
- China
- Prior art keywords
- mirna
- gem
- heart
- pharmaceutical composition
- disease
- 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.)
- Active
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a kind of GEM 132 medical composition and its uses of miRNA 2861.MiRNA 2861 antisense base sequences are 5 ' CCGCCCGCCGCCAGGCCCC 3 '；It is 10 that pharmaceutical composition, which includes infection titer,16It is cholesterol, nano particle or liposome that PFU, which is overexpressed the antisense nucleoside acid vectors of miRNA 2861, virus or auxiliary material, the carrier,；Viral vector is the one or more in adenovirus vector, slow virus carrier, retroviral vector；The auxiliary material is the one or more in mannitol, phosphate buffer, physiological saline.By oral or injectable in prevention and treatment myocardial hypertrophy, myocardial fibrosis, coronary heart disease and heart failure.Drug regimen composition formula of the present invention is reasonable, and pharmacology is reliable, and manufacture craft is simple, and therapeutic effect is obvious, has a wide range of application, and use environment is friendly.
Description
Technical field：
The present invention relates to a kind of endogenous non-coding tiny RNA s new drug and application thereof, especially a kind of miRNA-2861 is anti-
The medical composition and its use of adopted vaccination heart disease, belongs to biopharmaceutical technology.
Background technology：
Angiocardiopathy mainly includes hypertension, coronary heart disease and congestive heart failure etc., and angiocardiopathy is that the mankind are good for
The number one killer of health, there are 1,007 million peoples to die from angiocardiopathy every year in worldwide, its death rate is close to all
The summation of cancer mortality.As living standards of the people increasingly improve the change with dietary structure, the angiocardiopathy death rate is in
Obvious ascendant trend, turn into the first big cause of death more than cancer, prevention and treatment angiocardiopathy is still medical science and life
The significant task of thing.Myocardial hypertrophy refers to that horizontal upper the thickening for cardiac muscular tissue of tissue sets one's heart myocyte's volume with cellular level
The general name of increase is due to a kind of disease that cardiac muscle cell's volume increases and causes heart size increase to be occurred.The disease is by one
The Co stituation of a little physiology and pathological factor and occur, be the synthesis sex expression of many angiocardiopathies；Myocardial hypertrophy is the heart
One kind of myopathy, it is that cardiac muscle cell is directed to a kind of increased responsing reaction of Hemodynamics, a variety of situations can cause in human body
Hemodynamic increase, such as hypertension, heart valve disease.Long-term excess load hemodynamics, which stimulates, can cause with cardiac muscle cell
Cardiac muscle cell's remodeling process that volume increase is characterized, is exactly myocardial hypertrophy.Typically now think the cardiac muscle of two kinds of situations be present
Hypertrophy, one kind are normal physiologic hypertrophies, for example supporter develops the myocardial hypertrophy occurred after birth, and also physical training is drawn
The myocardial hypertrophy risen；Another is pathologic myocardial hypertrophy, and its early sign is that locular wall and interventricular septum thicken, myocardial contraction work(
It can strengthen, be accordingly regarded as compensatory hypertrophy, if the state of an illness is not eased always, the myocardial hypertrophy later stage can occur between ventricle wall
The exception of matter fibrosis, mycardial contractility functional disturbance and energetic supersession, gene expression and Electrophysiological characteristics, is ultimately resulted in
Cardiac failure.In view of myocardial hypertrophy is a progressive irreversible procedure, and eventually cause heart failure,
The landmark cardiac morphology that clinical lesion has occurred using it as heart for modern medicine changes, it is believed that it is to cause
One hazards of heart disease and cardiac sudden death.
With the upsurge of recent miRNA research applications, endogenous non-coding tiny RNA s adjusts as a gene expression
The center factor of section appears, and participates in many important physiology courses, and its target is determined for miRNA, the characteristics of control methods
Gene will not only have one, be one-to-many mode so that miRNA function is studied carefully abundant in content；MiRNA is for maintaining heart just
Normal physiological function plays an important roll, and the miRNA unconventionality expressions in heart are related to the generation of many heart diseases.Therefore, will
Target spots of the miRNA as cardiac disease treatment, exploitation related drugs have potential clinical value.；It is although more and more
MiRNA be found as the biomarker and therapy target of human diseases, but in cardiac hypertrophy and myocardial fibrosis heart
For crucial miRNA in disease still without determination, its function of being played is the challenge to field scientific research personnel.
The content of the invention：
It is an object of the invention to overcome the shortcomings of the prior art, seek to provide a kind of miRNA-2861 antisense nucleosides
Medical composition and its use of the acid for preparing diagnosis, prevention or treatment heart disease, it is determined that or discovery regulation and control cardiac muscle cell
The loose miRNA expressed in heart, further determines that its effect in heart disease, and be applied to these hearts
The diagnosis and preventing and treating of disease.
In order to realize foregoing invention purpose, the miRNA-2861 that the present invention designs antisense base sequences for 5 '-
CCGCCCGCCGCCAGGCCCC-3’；Pharmaceutical composition is made into using common process, including infection titer is 1016PFU is overexpressed
MiRNA-2861 antisense nucleosides acid vectors, virus or auxiliary material, the carrier are cholesterol, nano particle or liposome；Virus carries
Body is the one or more in adenovirus vector, slow virus carrier, retroviral vector；The auxiliary material is mannitol, phosphoric acid
One or more in salt buffer, physiological saline；
Pharmaceutical dosage form is the one or more in oral formulations, ejection preparation, tablet, therapy in dry powder form, is preferably injected
With small liquid drugs injection or injection freeze-dried powder.
The miRNA-2861 GEM 132 pharmaceutical compositions of the present invention, by oral or injectable in the prevention and treatment heart
Flesh plumpness, myocardial fibrosis, coronary heart disease and heart failure.
Pharmaceutical composition of the present invention expresses significantly up-regulation in the cardiac muscle cell of hypertrophy and cardiac hypertrophy, passes through structure
Build miRNA-2861 and be overexpressed the expression that adenovirus strengthens miRNA-2861, it is found that miRNA-2861 is overexpressed in cellular level energy
The loose generation of inducing cardiomyocytes, the expression of loose gene have obvious increase compared with control group；It is thin in the cardiac muscle of hypertrophy
It is significantly improved in born of the same parents compared with control group, the detection that loose index is carried out to it is found, the index such as cell table of myocardial hypertrophy
Area, the restructuring of albumen/DNA ratio and sarcomere also have obvious increase compared with control group, can induce myocardium fertilizer
Big generation, the biomarker as heart diseases such as early diagnosis and early prevention myocardial hypertrophy, myocardial fibrosis；To the heart
Flesh is plump and myocardial fibrosis has protective effect, is found in cellular level by transfecting miRNA-2861 GEM 132,
The cardiac myocyte hypertrophy that loose stimulating factor is induced can be suppressed, including loose index such as cell surface accumulates, albumen/DNA's
The restructuring of ratio and sarcomere also has obvious reduction compared with to stimulation group；In animal level by injecting external source miRNA-
The loose effect that 2861 GEM 132 can also suppress loose model includes cardiac phenotype, heart weight ratio, marginal zone
Cardiomyocytes cross-sectional area (WGA dyeing) and myocardial fibrosis (Masson dyeing) have obvious reduction, while heart function
Improve significantly.
Compared with prior art, its drug regimen composition formula is reasonable, and pharmacology is reliable, and manufacture craft is simple by the present invention, treatment
Positive effect, have a wide range of application, use environment is friendly.
Brief description of the drawings：
Fig. 1 is the cardiac muscle induced through cardiomegaly stimulating factor PE (phyenlephrinium) and Ang II (angiotensinⅡ)
The change of miRNA-2861 expressions during cellular mast；
Fig. 2 is to carry out PE processing to the primary cell transfected through miRNA-2861 GEM 132s, right on a cellular level
The loose suppression situation that PE is induced；
The flesh that Fig. 3 is cardiac muscle cell after carrying out PE processing to the primary cell transfected through miRNA-2861 GEM 132s is small
Save the result of the test schematic diagram of restructuring；
Fig. 4 can be suppressed the hypertrophy that PE induces by mouse mainline miRNA-2861 GEM 132s (antagomir)
The testing result schematic diagram of model；
Fig. 5 is that mouse mainline miRNA-2861 GEM 132s (antagomir) can suppress aortic coaractation (TAC)
The testing result schematic diagram of the mouse heart hypertrophy model induced.
Embodiment：
The present invention is made by way of example and in conjunction with the accompanying drawings and being expanded on further.
Embodiment 1,
MiRNA-2861 GEM 132s are overexpressed adenovirus vector and negative control adenovirus structure, the present embodiment use
Artificial synthesized miRNA-2861 antisense base sequences, subclone are connected into the pSilencer Adeno 1.0- of Ambion companies
CMV systems, structure miRNA-2861 GEM 132s are overexpressed adenovirus.Its miRNA-2861 GEM 132 nucleotide sequence
Such as following SEQ ID NO:Shown in 1：5’-CCGCCCGCCGCCAGGCCCC-3’
According to company's specification step, the carrier that Pac I is linearized and the adenoviral backbone of linearisation
(AdenovirTs LacZ Backbone) cotransfection HEK-293 cells, pack adenovirus, and virus, measure virus are collected in amplification
Titre；By empty carrier and adenoviral backbone according to above-mentioned steps cotransfection HEK-293 cells, negative control adenovirus is packed.
The recombinant adenovirus phosphate buffer of the overexpression miRNA-2861 GEM 132s of above-mentioned gained is diluted
For infection titer 1x1016PFU/ml, it is aseptic subpackaged in ampoule bottle by 1ml amounts, produce.
Embodiment 2,
The change of miRNA-2861 expressions in the case where PE and Ang II is stimulated, the present embodiment is for cardiac myocyte hypertrophy mould
Type, using the method culture rat suckling mouse primary cardiomyocytes established, (rat suckling mouse is purchased from Beijing Medical University, rat strains
For Wistar, the preparation of rat suckling mouse primary cardiomyocytes can be found in documents below：W.-Q.Tan,etal,Foxo3a
Inhibits Cardiomyocyte Hypertrophy through Transactivating Catalase J Biol
Chem.2008October 31；283(44):29730-29739), cardiac muscle cell is handled with PE and Ang II, trained
Foster different time, extracts the total serum IgE of cell, and Real-Time Fluorescent Quantitative PCR Technique detects miRNA-2861 expression, such as schemed
Shown in 1A, 1B, wherein, Figure 1A be rat suckling mouse primary cardiomyocytes after PE is handled miRNA-2861 with the time expression water
It is flat；Figure 1B be rat suckling mouse primary cardiomyocytes through Ang II processing after miRNA-2861 with the expression of time, ordinate
Represent on the basis of miRNA-2861 expression in untreated Neonatal Rat Primary Cardiomyocytes, it is treated in PE or Ang II
In journey in Neonatal Rat Primary Cardiomyocytes miRNA-2861 expression.MiRNA-2861 expressions are handled in PE and Ang II
There is significant rising within 6 hours.
Embodiment 3,
MiRNA-2861 GEM 132s can suppress the hypertrophy that PE is induced on a cellular level, and the present embodiment is to primary
After cardiac muscle cell transfects miRNA-2861 GEM 132s (anta-2861), it can effectively suppress miRNA-2861 expression, together
When cell cardiac muscle cell is handled with PE, and simultaneously use miRNA-2861 GEM 132s negative control (anta-NC)
Processing, after 24 hours, to cardiac muscle cell carry out myocardial hypertrophy index detection, as myocardial cell surface product (Fig. 2A),
Albumen/DNA (Fig. 2 B) detects, and the results show miRNA-2861 GEM 132s can effectively suppress the heart that PE is induced
Myocyte hypertrophy.
Embodiment 4,
MiRNA-2861 GEM 132s can suppress the weight for cardiac muscle cell's sarcomere that PE is induced on a cellular level
Group, the present embodiment transfect miRNA-2861 GEM 132s (anta-2861) and negative control (anta- to primary cardiomyocytes
NC after), cardiac muscle cell is handled with PE, after processing 24 hours, the restructuring that sarcomere is carried out to cardiac muscle cell is examined
Survey, as shown in figure 3, the results show miRNA-2861 GEM 132s can effectively suppress the cardiac muscle cell that PE is induced
Sarcomere restructuring.
Embodiment 5,
Mouse mainline miRNA-2861 GEM 132s (antagomir) can suppress the loose model that PE is induced,
After the present embodiment buries pump processing two weeks to mouse with PE, the heart of mouse has obvious loose phenotype, and adds miRNA- simultaneously
2861 GEM 132s (anta-2861), which can substantially weaken the loose effect that PE is induced, includes the cross-sectional area of cardiac muscle cell
Size (WGA dyeing) (such as Fig. 4 A) and heart weight ratio (such as Fig. 4 B)；Simultaneously miRNA-2861 GEM 132s are injected to mouse
Negative control (anta-NC) and PE, PE loose effect can not be weakened, experimental result confirms miRNA-2861 antisense nucleosides
Acid can suppress the generation for the cardiomegaly that PE is induced in integral level.
Embodiment 6,
Mouse mainline miRNA-2861 GEM 132s (antagomir) can suppress aortic coaractation (TAC) and be lured
The mouse heart hypertrophy model led, the present embodiment to mouse with aortic coaractation (Thoracic aorta constriction,
TAC method) can induce the generation of loose phenotype after handling three weeks；Add miRNA-2861 GEM 132s (anta- simultaneously
2861) the loose effect that TAC is induced can substantially be weakened；Add the negative control (anta- of miRNA-2861 GEM 132s
NC group) significantly changes the loose effect (Fig. 5) that can not weaken TAC and be induced, including the heart with TAC groups than no
The cross-sectional area size of dirty weight ratio (Fig. 5 A) and cardiac muscle cell (WGA (Wheat germ agglutinin) dyeing) (Fig. 5 B)；
Experimental result confirms that miRNA-2861 GEM 132s can suppress the generation for the cardiomegaly that TAC is induced in integral level.
。
Sequence table
SEQ ID NO:1
5’- CCGCCCGCCGCCAGGCCCC-3’
Claims (4)
1. a kind of miRNA-2861 GEM 132s pharmaceutical composition is preparing the purposes in being used to treat the medicine of heart disease,
It is characterized in that miRNA-2861 antisense base sequences are 5 '-CCGCCCGCCGCCAGGCCCC-3 ', the drug regimen
It is 10 that thing, which includes infection titer,16PFU is overexpressed miRNA-2861 antisense nucleosides acid vectors or virus, the carrier be cholesterol,
Nano particle or liposome；The virus is adenovirus vector, slow virus carrier, one kind in retroviral vector or more
Kind.
2. a kind of miRNA-2861 GEM 132s pharmaceutical composition according to claim 1 is being prepared for treating heart
Purposes in the medicine of disease, it is characterised in that the heart disease is myocardial hypertrophy, myocardial fibrosis, coronary heart disease or heart failure.
3. a kind of miRNA-2861 GEM 132s pharmaceutical composition according to claim 1 is being prepared for treating heart
Purposes in the medicine of disease, it is characterised in that the formulation of pharmaceutical composition is oral formulations or ejection preparation.
4. a kind of miRNA-2861 GEM 132s pharmaceutical composition according to claim 1 is being prepared for treating heart
Purposes in the medicine of disease, it is characterised in that the formulation of pharmaceutical composition is tablet or dry powder doses.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410803184.5A CN104491879B (en) | 2014-12-19 | 2014-12-19 | A kind of GEM 132 medical composition and its uses of miRNA 2861 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410803184.5A CN104491879B (en) | 2014-12-19 | 2014-12-19 | A kind of GEM 132 medical composition and its uses of miRNA 2861 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104491879A CN104491879A (en) | 2015-04-08 |
| CN104491879B true CN104491879B (en) | 2017-12-12 |
Family
ID=52933390
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410803184.5A Active CN104491879B (en) | 2014-12-19 | 2014-12-19 | A kind of GEM 132 medical composition and its uses of miRNA 2861 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104491879B (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103476947A (en) * | 2011-03-02 | 2013-12-25 | 格路福生物制药公司 | Enhanced biodistribution of oligomers |
| US20140087964A1 (en) * | 2012-09-24 | 2014-03-27 | University Of Virginia Patent Foundation | Compositions and methods for detecting aberrant regulation, expression, and levels of hgh |
-
2014
- 2014-12-19 CN CN201410803184.5A patent/CN104491879B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104491879A (en) | 2015-04-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104548135A (en) | Medicinal composition containing LncRNA and application thereof | |
| CN102886050B (en) | Application of miRNA-489 and medicinal composition | |
| CN101643791A (en) | MicroRNA-328 and application of antisense nucleotide thereof for diagnosing, preventing and curing heart diseases | |
| Huang et al. | Cardiac-specific Traf2 overexpression enhances cardiac hypertrophy through activating AKT/GSK3β signaling | |
| CN104548136B (en) | A kind of piRNA medical composition and its uses | |
| CN104645352B (en) | A kind of piRNA GEM 132s medical composition and its use | |
| CN102115787A (en) | MicroRNA (Ribose Nucleic Acid) and application of antisensenucleic acid of microRNA to diagnosis, prevention, treatment and/or prognostic evaluation of heart disease | |
| Elton et al. | MicroRNAs in cardiovascular disease | |
| CN105535993A (en) | Use of recombinant oncolytic vaccinia virus in treatment of gastric cancer | |
| CN102242080B (en) | Method for treating or diagnosing heart failure or tendency of heart failure or improving functions of myocardial cells by miR-24 (MicroRNA-24) | |
| CN106222173A (en) | CircRNA MNCR nucleotide, medical composition and its use containing this nucleotide | |
| CN106138081A (en) | A kind of medical composition and its use of targeting circRNA | |
| CN106222174A (en) | CircRNA CHIF nucleotide, containing the medical composition and its use of this nucleotide | |
| CN107184594A (en) | The 3p of miR 142 are used for composition and the application for preventing and/or treating heart disease | |
| CN104491879B (en) | A kind of GEM 132 medical composition and its uses of miRNA 2861 | |
| CN102266570A (en) | New application of miRNA-484, pharmaceutical composition containing miRNA-484 and use thereof | |
| WO2022135278A1 (en) | Novel application of azd3965 medicine | |
| CN104524599A (en) | Antisense nucleotide MiRNA-532 containing pharmaceutical composition and application thereof | |
| CN104491878A (en) | Pharmaceutical composition containing miRNA-324 and application of pharmaceutical composition | |
| CN102251037A (en) | Application of miRNA-23a antisense nucleotide in preparation of medicine for treating heart diseases | |
| CN105969735A (en) | Ischemia hypoxia-resistant human mesenchymal stem cells, and preparation method and application | |
| CN102973953A (en) | MiRNA (micro ribose nucleic acid)-874 and application of miRNA-874antisense nucleotide | |
| CN110229879A (en) | Its product of the application and application of a kind of piRNA-500 nucleotide analog and its GEM 132 | |
| CN104988151A (en) | Application of miRNA-9-5p compound in preparing chronic pain diagnosis marker and treatment drugs | |
| CN1219054C (en) | Structure for recombinant adenovirus with double killer function and application in tumor treatment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Li Peifeng Inventor after: Wang Kun Inventor after: Zhou Luyu Inventor before: Wang Kun Inventor before: Li Peifeng |
|
| CB03 | Change of inventor or designer information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |