CN106138081A - A kind of medical composition and its use of targeting circRNA - Google Patents

A kind of medical composition and its use of targeting circRNA Download PDF

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CN106138081A
CN106138081A CN201610662639.5A CN201610662639A CN106138081A CN 106138081 A CN106138081 A CN 106138081A CN 201610662639 A CN201610662639 A CN 201610662639A CN 106138081 A CN106138081 A CN 106138081A
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mfacr
injection
circrna
sirna
myocardial
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CN106138081B (en
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王昆
李培峰
周露玙
刘翠云
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Qingdao University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A61K9/0056—Mouth soluble or dispersible forms; Suckable, eatable, chewable coherent forms; Forms rapidly disintegrating in the mouth; Lozenges; Lollipops; Bite capsules; Baked products; Baits or other oral forms for animals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions

Abstract

The invention discloses the pharmaceutical composition of a kind of targeting circRNA, it includes MFACR siRNA nucleotide, pharmaceutically acceptable carrier or viral vector and adjuvant；Described carrier is one or more in chitosan, cholesterol, nano-particle and liposome, it is preferably liposome, described adjuvant is one or more in mannitol, phosphate buffer, normal saline, it is preferably phosphate buffer, described MFACR siRNA content is 0.8 1.5 micrograms, and MFACR siRNA nucleotide and liposome mass ratio are 1:1.25；Carrier and adjuvant mass ratio are 1:200.Pharmaceutical composition, for myocardial infarction, treating myocardial ischemia damage, myocardial hypertrophy and the treatment of myocardial fibrosis and preventing and treating, is administered with oral or injection system；Described injection is intravenous injection, intramuscular injection, intracoronary injection or direct myocardial injection.Compared with prior art, its raw material selected is scientific and reasonable, and preparation technology is simple, and medicine effect is obvious, and range is wide, safe and reliable, and applied environment is friendly.

Description

A kind of medical composition and its use of targeting circRNA
Technical field:
The invention belongs to biomedicine technical field, relate to nucleic acid new drug and the purposes of a kind of targeting endogenous circRNA, A kind of MFACR-siRNA nucleotide pharmaceutical composition containing targeting circRNA MFACR and at prevention or the treatment heart Purposes in dirty disease.
Background technology:
Cardiovascular disease is the number one killer of human health, and the whole world has more than 1,700 ten thousand people to die from cardiovascular disease every year Disease, its mortality rate, close to the summation of all cancer mortalities, is the chief threat of human health and life.Along with human lives Level improves the change with dietary structure day by day, and cardiovascular disease mortality rate is obvious ascendant trend, and having exceeded cancer becomes The one big cause of death.Recent statistics result according to announcing in the recent period shows, 18 years old and above prevalence of hypertension rate are 18.8%, estimate whole nation number of patients more than 1.6 hundred million, myocardial hypertrophy, myocardial infarction, coronary heart disease and the heart failure thereby resulted in The patient apoptosis related cardiac conditions such as exhausting reaches several ten million, and the whole world is died from the number of cardiovascular and cerebrovascular disease every year and is up to 15,000,000 People, occupies the various cause of the death the first.Be not fully understood about heart disease pathogeny at present, the prevention of heart disease, diagnosis and Treatment still can not reach satisfactory effect, is badly in need of the new technical method of exploitation for cardiopathic diagnosis and preventing and treating.
SiRNA is a kind of small RNA molecular (about 21-25 nucleotide), is make gene silencing the most powerful and strong Biological tool, it is possible to the silence of the target gene that induced sequence is special, rapid blocking gene activity.Calendar year 2001, research worker is sent out The siRNA now synthesized, can induce the RNAi effect in mammal body, the activity of suppression target gene, have application greatly It is worth.Recently research finds that annular RNA (circRNA) the most also has important function.CircRNA is that a class is special Non-coding RNA molecule, is generally produced by special alternative splicing, is present in a large number in the Cytoplasm of eukaryotic cell.circRNA Molecule is closed circular structure, it is possible to be combined with cellular endogenous miRNA, plays miRNA sponge (miRNA in cell Sponge) effect, thus corresponding function is exercised in the expression regulating miRNA.
Apoptosis (apoptosis), also known as programmed cell death, is that phalangeal cell is in certain physiology or pathological conditions Under, it then follows the program of self, the process of oneself end lives, be an active, high-sequential, by Gene Handling and one be The process that row enzyme participates in, to normal embryo development, all plays an important role during maintaining cell colony and malignant change, is ensureing The healthy survival processes of multicellular organism plays the role of key.Under many myocardial damages or heart pathological state, as Myocardial ischemia reperfusion injury, myocardial hypertrophy and heart failure etc., myocardial cell can occur apoptosis.Due to ripe myocardial cell Can not division growth, myocardial cell excessive Apoptosis will necessarily make myocardial cell number reduce, this be probably heart disease morbidity Mechanism.Also do not have research to disclose circRNA to play a role during apoptosis of cardiac muscle, find specifically expressing in heart, Participate in the circRNA of apoptosis of cardiac muscle, illustrate its mechanism of action, for developing the siRNA of targeting circRNA at heart disease Diagnosis and treatment in terms of application have and important meaning and vast prospect.
Our research finds circRNA MFACR when of myocardial cell generation apoptosis, and full name is mitochondrial function phase The annular RNA closed, expresses notable rise, and circRNA MFACR has the effect promoting apoptosis of cardiac muscle.Use corresponding SiRNA (i.e. MFACR-siRNA) can targeted silent circRNA MFACR, thus suppress the cardiac muscle in During Myocardial Infarction Process thin Born of the same parents' apoptosis problem, finally alleviates the injury that heart is caused by infarction.
Summary of the invention:
It is an object of the invention to the shortcoming overcoming prior art to exist, according to the heart that can regulate and control apoptosis of cardiac muscle MFACR-siRNA that specificity is expressed and fine at myocardial ischemia-reperfusion, myocardial infarction, coronary heart disease, myocardial hypertrophy and cardiac muscle Purposes in the heart diseases such as dimensionization, a kind of new pharmaceutical composition of exploitation, for the diagnosis and treatment of heart disease.
In order to realize foregoing invention purpose, the MFACR-siRNA nucleotide of the regulating cell apoptosis that the present invention relates to, used The sequence of MFACR-siRNA nucleotide be 5 '-AAGACAUGCCCUGUGUGAC-3 ', i.e. SEQ ID NO:1.
The pharmaceutical composition that the present invention relates to includes MFACR-siRNA nucleotide, pharmaceutically acceptable carrier and adjuvant； Described carrier is one or more in chitosan, cholesterol, nano-particle and liposome, it is therefore preferable to liposome, described auxiliary Material is one or more in mannitol, phosphate buffer, normal saline, it is therefore preferable to phosphate buffer, described MFACR- SiRNA content is 0.8-1.5 microgram, and MFACR-siRNA nucleotide and liposome mass ratio are 1:1.25；Carrier and adjuvant quality Ratio is 1:200.
The another kind of pharmaceutical composition that the present invention relates to includes MFACR-siRNA nucleotide, viral vector and adjuvant；Virus Carrier is one or more in adenovirus vector, slow virus carrier, retroviral vector, it is therefore preferable to adenovirus vector, Described adjuvant is one or more in mannitol, phosphate buffer, normal saline, it is therefore preferable to phosphate buffer, described The adenovirus vector infection titer of MFACR-siRNA is 1016PFU。
The medicine composition dosage form that the present invention relates to be the one in oral formulations, ejection preparation, tablet, therapy in dry powder form or Multiple, preferably injection freeze-dried powder.
The pharmaceutical composition that the present invention relates to is for myocardial infarction, treating myocardial ischemia damage, myocardial hypertrophy and myocardial fibrosis Treatment and preventing and treating, be administered with oral or injection system；Described injection is intravenous injection, intramuscular injection, intracoronary injection Or directly myocardial injection, preferably intravenous injection.
The present invention also provides for a kind of test kit for preventing or treat heart disease, containing the composition of MFACR-siRNA Medicine imports internal, for prevention and the treatment of myocardial infarction, treating myocardial ischemia damage, myocardial hypertrophy and myocardial fibrosis.
The present invention is found through experiments circRNA MFACR at treating myocardial ischemia damage and the apoptosis of cardiac muscle of hypoxia inducible During express notable rise, the expression of circRNA MFACR can be lowered by transfection and injection MFACR-siRNA, Suppression apoptosis of cardiac muscle, treating myocardial ischemia damage and myocardial infarction area reduce.Compared with prior art, its raw material section selected Learning rationally, preparation technology is simple, and medicine effect is obvious, and range is wide, safe and reliable, and applied environment is friendly.
Accompanying drawing illustrates:
Fig. 1 be anoxic treatment (1A) and treating myocardial ischemia damage (1B) apoptosis of cardiac muscle during circRNA MFACR The variation diagram of expression, wherein the rna expression level of MFACR levels:circRNA MFACR；Control: matched group； A/R:Anoxia/reoxygenation, refers to process primary cardiomyocytes hypoxia-reoxygenation；Sham: sham operated rats, for I/R group Negative control；I/R:ischaemia/reperfusion, refers to mice is carried out Ischemia Reperfusion operation.
Fig. 2 be transfection MFACR-siRNA adenovirus after anoxic treatment apoptosis of cardiac muscle during chondriokinesis (2A) and the variation diagram of Level of Apoptosis (2B), wherein there is line in Fragmented mitochondria (%of cells) The cell of plastochondria division is the percentage ratio of total cell in accounting for a visual field；A/R:Anoxia/reoxygenation, refers to the primary heart Myocyte's hypoxia-reoxygenation processes；The out of order comparison of MFACR-sc:MFACR-siRNA, can suppressing of MFACR-siRNA: design The siRNA fragment of MFACR.
Fig. 3 is chondriokinesis and thin during the apoptosis of cardiac muscle for the treatment of myocardial ischemia damage after injection MFACR-siRNA The variation diagram of born of the same parents' level of apoptosis, wherein Fission (%of mitochondria): refer to that occurring the mitochondrion of division to account for observes The percentage ratio of total mitochondria number；Apoptotic cells (%): refer to occur the percentage ratio shared by cell of apoptosis；Sham: Sham operated rats, for the negative control of I/R group；I/R:ischaemia/reperfusion, refers to mice is carried out Ischemia Reperfusion hands Art；The out of order comparison of MFACR-sc:MFACR-siRNA, the siRNA fragment that MFACR can be suppressed of MFACR-siRNA: design.
Fig. 4 is the result schematic diagram that MFACR-siRNA reduces treating myocardial ischemia damage area, wherein %LV or AAR: represent Left ventricle Infarct area and the percentage ratio of dangerous area；AAR/LV: hazardous area/left ventricle；INF/AAR: infarcted region/danger District.Sham: sham operated rats, for the negative control of I/R group；I/R:ischaemia/reperfusion, refers to mice is carried out ischemia Fill operation again；The out of order comparison of MFACR-sc:MFACR-siRNA, MFACR-siRNA: design can suppress MFACR's SiRNA fragment.
Detailed description of the invention:
Below by specific embodiment and combine accompanying drawing the present invention is expanded on further.
Material, experimental technique and the operating procedure related in following embodiment, including primary myocardial cell culture, cell Transfection, myocardial ischemia and reperfusion operating procedure, the double dye of Evans blue/TTC detects with TUNEL and all sees documents below: (1) Wang JX,et al,miR-499regulates mitochondrial dynamics by targeting calcineurin and dynamin-related protein-1.2011January 17:71-78；(1)Wang K,Liu CY,Zhang XJ,Feng C,Zhou LY,Zhao Y,Li PF.Mir-361-regulated prohibitin inhibits mitochondrial fission and apoptosis and protects heart from ischemia injury.Cel l death and differentiation.2015；22:1058-1068.Experimental animal wherein used is mice, and its mouse strain is C57/ BL6, purchased from Beijing Medical University；Used enzyme such as restricted enzyme, expand enzyme and reverse transcriptase etc., be purchased from Beijing extensive and profound in meaning Biological company limited；Reagent used is analytical pure level reagent, and obtains from commercially available channel.
Embodiment 1: circRNA expression during the apoptosis of cardiac muscle of myocardial ischemia and Myocytes Anoxia induction
The present embodiment uses apoptosis of cardiac muscle experimental model, cultivates the primary cardiac muscle of rat neonatal rat by conventional method thin Born of the same parents, hypoxia incubator (oxygen concentration is less than 1%) is cultivated different time, is extracted RNA, Real-Time Fluorescent Quantitative PCR Technique detection The expression of circRNA MFACR, as shown in Figure 1A in hypoxia processes 4h-8h, the when of myocardial cell generation apoptosis CircRNA MFACR significantly raises, and wherein Control is A/R matched group；
The present embodiment uses Banded Rats coronary artery to set up myocardial infarction and ischemia model, and ischemia different time is cored dirty ischemic region And non-ischemic region cardiac muscular tissue, extract total serum IgE, by the expression water of Real-Time Fluorescent Quantitative PCR Technique detection circRNA MFACR Flat, as shown in Figure 1B: more non-ischemia model Zu Fei ischemic tissue of ischemia 120min myocardial infarction and ischemia model Myocardial ischemic tissue CircRNA MFACR expression significantly rises, and wherein using sham operated rats (sham) as I/R matched group, and then proves CircRNA MFACR plays a role in apoptosis process.
The apoptosis of cardiac muscle of embodiment 2:MFACR-siRNA suppression hypoxia inducible
The myocardial cell using original cuiture in the present embodiment is model, to the primary cardiomyocytes transfection transfection cultivated MFACR-siRNA adenovirus vector, after transfecting 24 hours, in hypoxia incubator, cultivation processes cell 3 hours, and myocardial ischemia lures Guided cell apoptosis, detects apoptosis of cardiac muscle situation with cell apoptosis detection kit (TUNEL method).The four of contrast different disposal Group myocardial cell (as shown in fig. 2 a or 2b, is from left to right followed successively by and does not has the myocardial cell group of anoxic treatment；Anoxic treatment cardiac muscle Groups of cells, anoxic treatment myocardial cell group after transfection MFACR-sc adenovirus vector, i.e. negative control group, transfect MFACR- Anoxic treatment myocardial cell group after siRNA adenovirus vector) Mitochondria division and the change of Level of Apoptosis, can from Fig. 2 A To obtain the myocardial cell chondriokinesis level of hypoxia inducible after primary cardiomyocytes transfection MFACR-siRNA adenovirus vector Declining, after Fig. 2 B explanation transfection MFACR-siRNA adenovirus vector, the apoptosis of cardiac muscle level of hypoxia inducible declines, i.e. MFACR-siRNA effectively suppresses the apoptosis of cardiac muscle of hypoxia inducible.
The apoptosis of cardiac muscle of embodiment 3:MFACR-siRNA suppression treating myocardial ischemia damage
The present embodiment, with C57/BL6 mice as experimental subject, often organizes 8 mices, result one-way that will record ANOVO statistical software carries out statistical analysis, the most respectively injection 30mg/kg MFACR-siRNA and its feminine gender Compareing MFACR-sc, mouse weights is also anaesthetized, and dorsal position is fixed, and cropping also uses iodine disinfection field of operation.Cervical region medisection gas Managing and intubate, even animal respirator carries out positive airway pressure, longitudinal incision skin about 3cm at left border of sternum and at heartbeat, Successively blunt separation subcutaneous tissue, muscle, open breast, carefully mentions pericardium and cuts off, and fully exposes heart；Dilute MFACR-respectively SiRNA and MFACR-sc is in phosphate buffer, and final volume is 200 μ L, and No. 26 conduits enter tremulous pulse root, injection from the apex of the heart Time press from both sides simultaneously and close aorta and pulmonary artery, persistently folder closes 20 seconds, monitors 5min, treats that heart recovers Dou Lvhou, removes thoracic cavity inner product Blood also spends needle applicator intake-gas closedown thoracic cavity, after waiting mice clear-headed, separating respiratory machine, puts back in cage；Electrocardiogram is supervised Survey whole operation process；Carrying out heart ischemia reperfusion operation after injecting 3 days, concrete operations are as follows: after mouse anesthesia, and back of the body position is solid On experiment plank, and carry out electrocardiogram monitoring.Cervical region medisection trachea also intubates, and interlocks thing artificial respirator, in 4th intercostal opens breast, carefully mentions pericardium and cuts off, and fully exposes heart, blood vessel；Between pulmonary conus and left auricle, with Great cardiac vein trunk is mark, inserting needle at 2mm below left auricle root, depth of needle 0.5mm；With 6/0 band pin stitching thread Through cardiac muscle top layer, at pulmonary artery cone branch pin；After electrocardiogram recovers to stablize 10min, before being ligatured left coronary artery Descending branch (LAD)；Using I, aVL ST-Segment back of a bow upwards raise more than 0.1mv and persistently more than 0.5h as ligation successfully mark Will, after ischemia 120min, unclamps ligature and starts Reperfu-sion, drops to mark with ST section；Remove in thoracic cavity hematocele and spend Needle applicator intake-gas closes thoracic cavity；Reperfu-sion uses TUNEL detection kit (Roche company) by former after 24 hours Position end gap labelling method (TUNEL method) detection apoptosis of cardiac muscle, under the conditions of different disposal, apoptotic percentage is such as schemed Shown in 3B, result shows that MFACR-siRNA can suppress the apoptosis of cardiac muscle for the treatment of myocardial ischemia damage.Take MFACR-siRNA simultaneously Group mouse heart carries out ultrathin section, and Electronic Speculum detects mitochondrial splitting degree, and size is less than 0.6mm2For division line grain Body, under the conditions of different disposal, mitochondrial splitting degree is as shown in Figure 3A, and result display MFACR-siRNA can significantly inhibit cardiac muscle Myocardial cell chondriokinesis that ischemia-reperfusion causes and apoptosis.
The myocardial infarction area of embodiment 4:MFACR-siRNA suppression treating myocardial ischemia damage
Building the method such as embodiment 3 of mice ischemia-reperfusion model in the present embodiment, in Fig. 4 result, ARR/LV represents dangerous District's gross area/left ventricular area, INF/AAR represents Infarct area/hazardous area gross area, INF/LV represent Infarct area/ Left ventricular area, the most often organizes and is from left to right followed successively by ischemia-reperfusion sham operated rats (Sham), through ischemia-reperfusion group (I/ R), ischemia-reperfusion group (I/R+MFACR-sc) and inject ischemia-reperfusion group (I/ after MFACR-siRNA after injection MFACR-sc R+MFACR-siRNA), result shows: the mice I/R group myocardial infarction area of injection MFACR-siRNA is brighter than MFACR-sc group Aobvious reduction, has significant difference (p < 0.01).The myocardial infarct size that the present embodiment relates to calculates as follows: left ventricular area (left ventricle, LV) is blue, red and white area sum；The hazardous area gross area (area at risk, AAR) is Red and white area sum；Infarct area (infarct area, INF) is white area, dangerous area (AAR/ LV, %)=(the hazardous area gross area/left ventricular area) × 100%, myocardial infarct size (INF/AAR, %)=(Infarct area/danger The danger zone gross area) × 100%, above result explanation mice underwent coronary injection MFACR-siRNA can significantly inhibit cardiac muscle Ischemical reperfusion injury.
Sequence table
SEQ ID NO:1:
5’- AAGACAUGCCCUGUGUGAC -3’

Claims (8)

1. the pharmaceutical composition of a targeting circRNA, it is characterised in that include MFACR-siRNA nucleotide, pharmaceutically can connect The carrier being subject to and adjuvant；Described carrier is one or more in chitosan, cholesterol, nano-particle and liposome, described auxiliary Material is one or more in mannitol, phosphate buffer, normal saline.
The pharmaceutical composition of targeting circRNA the most according to claim 1, it is characterised in that described carrier is liposome, Described adjuvant is phosphate buffer, and described MFACR-siRNA content is 0.8-1.5 microgram, MFACR-siRNA nucleotide and lipid Body mass ratio is 1:1.25；Carrier and adjuvant mass ratio are 1:200.
3. the pharmaceutical composition of a targeting circRNA, it is characterised in that include MFACR-siRNA nucleotide, viral vector and Adjuvant；Viral vector is one or more in adenovirus vector, slow virus carrier, retroviral vector, and described adjuvant is One or more in mannitol, phosphate buffer, normal saline, the adenovirus vector of described MFACR-siRNA infects and drips Degree is 1016PFU。
The pharmaceutical composition of targeting circRNA the most according to claim 3, it is characterised in that viral vector is adenovirus Carrier, described adjuvant is phosphate buffer.
5. according to the pharmaceutical composition of the targeting circRNA described in any one of claim 1-4, it is characterised in that described MFACR-siRNA nucleotides sequence is classified as 5 '-AAGACAUGCCCUGUGUGAC-3 ', i.e. SEQ ID NO:1.
The pharmaceutical composition of targeting circRNA the most according to claim 5, it is characterised in that medicine composition dosage form is One or more in oral formulations, ejection preparation, tablet, therapy in dry powder form, preferably injection freeze-dried powder.
The pharmaceutical composition of targeting circRNA the most according to claim 6, it is characterised in that medicine composition dosage form is Injection freeze-dried powder.
The pharmaceutical composition of targeting circRNA the most according to claim 7, it is characterised in that pharmaceutical composition is used for the heart The treatment of flesh infarction, treating myocardial ischemia damage, myocardial hypertrophy and myocardial fibrosis and preventing and treating, be administered with oral or injection system；Institute Stating injection is intravenous injection, intramuscular injection, intracoronary injection or direct myocardial injection.
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CN115961025A (en) * 2020-06-30 2023-04-14 苏州大学附属第二医院 Application of circular RNA in preparation of products related to myocardial hypertrophy

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CN113136427A (en) * 2020-01-17 2021-07-20 中国科学技术大学 Application of mitochondrial circular RNA as hepatocellular carcinoma biomarker
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CN115961025B (en) * 2020-06-30 2023-07-18 苏州大学附属第二医院 Application of circular RNA in preparation of products related to myocardial hypertrophy

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