[go: up one dir, main page]

WO2012017105A1 - Micro-arn utilisé pour le traitement de canalopathies arythmogènes - Google Patents

Micro-arn utilisé pour le traitement de canalopathies arythmogènes Download PDF

Info

Publication number
WO2012017105A1
WO2012017105A1 PCT/ES2011/000248 ES2011000248W WO2012017105A1 WO 2012017105 A1 WO2012017105 A1 WO 2012017105A1 ES 2011000248 W ES2011000248 W ES 2011000248W WO 2012017105 A1 WO2012017105 A1 WO 2012017105A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
arrhythmogenic
polynucleotide
sequence
mir
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.)
Ceased
Application number
PCT/ES2011/000248
Other languages
English (en)
Spanish (es)
Inventor
Diego Franco Jaime
Amelia Eva Aranega Jimenez
Houría DAIMI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universidad de Jaen
Original Assignee
Universidad de Jaen
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidad de Jaen filed Critical Universidad de Jaen
Publication of WO2012017105A1 publication Critical patent/WO2012017105A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical

Definitions

  • the present invention falls within the field of gene therapy and cardiology and specifically refers to the use of the hsa-mir-219-5p microRNA, of SEQ ID NO: 1, of the modified polynucleotides derived therefrom, SEQ ID NO: 3 or SEQ ID NO: 5, or of its precursors, SEQ ID NO: 2, SEQ ID NO: 4 or SEQ ID NO: 6, for the preparation of useful medicines for the treatment of arrhythmogenic canalopathies, preferably those that are due to alterations in the function of the cardiac sodium channel, more preferably of type 3 long QT syndrome and Brugada syndrome.
  • Ventricular arrhythmias are the most frequent cause of sudden death in developed societies and are usually related to the existence of ischemic heart disease, of which they may be the first manifestation in hitherto asymptomatic subjects. Less frequently it occurs in subjects without structural heart disease and younger. Advances in molecular biology in the last decade have allowed the identification of several clinical entities that share their origin in mutations in genes that code for subunits of ion channels, which are responsible for generating the potential for action of heart cells. This set of inherited diseases are called arrhythmogenic canalopathies. In recent years, much progress has been made in the knowledge of the genetic and functional basis of different arrhythmogenic syndromes, including Brugada syndrome and long QT syndrome.
  • Brugada syndrome is a canalopathy associated with abnormalities in the sodium channels, specifically mutations in the SCN5A gene that codes for the transmembrane protein Nav1 .5, which in all cases cause a lack of function of this channel and usually occur associated with polymorphic ventricular arrhythmias in subjects without heart disease. These subjects they have a characteristic electrocardiogram: ST segment elevation in the right precordial leads and blocking pattern of the right bundle of the His bundle, usually with normal QT.
  • microRNAs are small-sized single-stranded RNA molecules (approximately 22 nucleotides) and non-coding, which bind to specific sequences of the non-coding regions (mainly the 3 ' UTRs) of the target messenger RNAs, inhibiting their translation and / or destabilizing them, which also causes a decrease in the total amount of protein.
  • target transcripts encoding cardiac ion channel proteins for example, microRNA-195 and microRNA-1 act on the expression of the SCN5A gene (Baofeng Yang, et al., 2008, Cardiovascular Research, Vol. 79: 571-580).
  • the present invention proposes the use of the hsa-mir-219-5p microRNA, of SEQ ID NO: 1, of the modified polynucleotides derived therefrom, SEQ ID NO: 3 or SEQ ID NO: 5, or of its precursors, SEQ ID NO: 2, SEQ ID NO: 4 or SEQ ID NO: 6, for the preparation of medicines useful for the treatment of arrhythmogenic canalopathies, preferably those that are due to alterations in the function of the cardiac sodium channel, more preferably of syndrome of long QT type 3 and Brugada syndrome.
  • the small size of the microRNAs their easy manipulation, the bioavailability of their precursors (pre-miR or pre-microRNA) and their high thermodynamic stability, make them molecules with a high therapeutic potential.
  • the conjugation of the polynucleotides of the invention with, for example, but not limited to lipid residues, allows their bioavailability in the organism of the patient to which they are administered.
  • polynucleotide sequences of the invention or polynucleotides of the invention allow modulating the function of the cardiac sodium channel through its binding to the mRNA encoding the Nav1.5 protein, or in the case of precursors, providing the polynucleotide that binds to this mRNA, which gives the possibility of increase the function of the sodium channel in patients with, for example, but not limited to, Brugada syndrome, or decrease it in patients with, for example, but not limited to, LQT3 syndrome, and thus reverse the altered biological function in these pathologies cardiac, as opposed to current biomechanical or pharmacological methods that only partially mitigate the symptoms of these diseases.
  • the inventors demonstrate that the hsa-mir-219-5p microRNA, or miR-219a, of SEQ ID NO: 1, is capable of reducing the expression of the SCN5A gene, which codes for the Nav1.5 transmembrane protein responsible for the potential of action in cardiac muscle cells.
  • SCN5A gene which codes for the Nav1.5 transmembrane protein responsible for the potential of action in cardiac muscle cells.
  • In vitro analysis of this microRNA shows that its overexpression decreases the expression of the SCN5A gene up to 50% in atrial cardiac muscle cells ( Figure 3), thus reducing its capacity and contractile rhythm.
  • the present invention proposes the use of this microRNA for the preparation of medicaments for the treatment of arrhythmogenic canalopathies, preferably of those arrhythmogenic canalopathies whose cause is alterations in the sodium channels, more preferably when the alterations in said channels are due to function gain mutations in the SCN5A gene, such as, but not limited to, the LQT3 syndrome.
  • the three nucleotides (CAA) adjacent to the 3 " end of the GAUUGUC sequence or complementary sequence to the mRNA of the SCN5A gene comprised in SEQ ID NO: 1, have been replaced by three other nucleotides (AGC), of so that SEQ ID NO: 3 has been obtained, a polynucleotide that has at least 85% identity with SEQ ID NO: 1 and that comprises three complementary sites more than this sequence with the mRNA of the SCN5A gene, so it has a greater binding capacity to it than the sequence of the native microRNA or SEQ ID NO: 1.
  • SEQ ID NO: 3 is also useful for the preparation of medicaments for the treatment of arrhythmogenic canalopathies, preferably of those arrhythmogenic canalopathies whose cause they are alterations in the sodium channels, more preferably when the alterations in said channels are due to function gain mutations in this gene, such as, but not limited to, the sin drome of LQT3.
  • a first aspect of the invention relates to the use of an isolated polynucleotide, hereinafter "polynucleotide of the invention", which comprises a nucleotide sequence with at least 60%, preferably with at least 75%, more preferably with at least 80% and even more preferably with at least 85% identity with respect to the full length of the sequence SEQ ID NO: 1 for the preparation of a medicament, or alternatively, to an isolated polynucleotide comprising a nucleotide sequence with at least 60%, preferably with at least 75%, more preferably with at least 80% and even more preferably with at least 85% identity with respect to the full length of the sequence SEQ ID NO: 1 for use as a medicine.
  • polynucleotide of the invention which comprises a nucleotide sequence with at least 60%, preferably with at least 75%, more preferably with at least 80% and even more preferably with at least 85% identity with respect to the full length of the sequence SEQ ID NO: 1 for use as
  • nucleotide sequence refers to a polymeric form of nucleotides of any length that may or may not be, chemically or biochemically modified. Thus, they refer to any polyiribonucleotide or polydeoxyribonucleotide , both single chain and double strand.
  • the polynucleotide of the invention can be obtained artificially by conventional cloning and selection methods, or by sequencing.
  • the polynucleotide of the invention comprises the nucleotide sequence SEQ ID NO: 3, which has at least 85% identity with respect to the full length of SEQ ID NO: 1.
  • the polynucleotide of the invention is SEQ ID NO: 4, a precursor nucleotide sequence of SEQ ID NO: 3 comprising this sequence.
  • SEQ ID NO: 5 a polynucleotide, SEQ ID NO: 5, which has at least 85% identity with SEQ ID NO: 1 and comprising three complementary sites less than this sequence with the mRNA of the SCN5A gene, so it has a lower binding capacity than the sequence of the native microRNA or SEQ ID NO: 1.
  • the SEQ ID NO: 5 is also useful for the preparation of drugs for the treatment of arrhythmogenic canalopathies, preferably of those arrhythmogenic canalopathies whose cause is alterations in the sodium channels, more preferably when the Alterations in these channels are due to function gain mutations in this gene, such as, but not limited to, LQT3 syndrome, since, although this sequence has fewer complementary sites than SEQ ID NO: 1 with the target mRNA It is also able to bind to said mRNA and reduce its function;
  • These medications may be useful, for example, but not limited to, in patients with LQT3 syndrome in which there is a deficiency in the expression of the native microRNA.
  • the polynucleotide of the invention comprises the nucleotide sequence SEQ ID NO: 5, which has at least 85% identity with respect to the full length of SEQ ID NO: 1.
  • the polynucleotide of the invention is SEQ ID NO: 6, a precursor nucleotide sequence of SEQ ID NO: 5 comprising this sequence.
  • the polynucleotide of the invention comprises a nucleotide sequence with at least 90% identity with respect to the full length of the sequence SEQ ID NO: 1.
  • the polynucleotide of the invention comprises a nucleotide sequence with at least 95% of identity with respect to the full length of the sequence SEQ ID NO: 1.
  • the polynucleotide of the invention comprises a nucleotide sequence with at least 98% identity with respect to the full length of the sequence SEQ ID NO: 1.
  • the polynucleotide of the invention comprises the nucleotide sequence SEQ ID NO: 1.
  • the polynucleotide of the invention is SEQ ID NO: 2, a precursor nucleotide sequence of SEQ ID NO: 1 comprising this sequence. Also within the scope of this invention are precursor nucleotide sequences or precursors.
  • precursor or “precursor sequence” as used herein includes any nucleotide sequence that when processed by, for example, but not limited to, enzymatic cleavage, is capable of providing a polynucleotide with at least 60%, preferably with at least 65%, more preferably with at least 70%, with at least 75%, with at least 80%, with at least 85%, with at least 90%, with at least 95% or with at least 98% identity with respect to the full length of SEQ ID NO: 1, or that it is capable of providing a sequence polynucleotide of SEQ ID NO: 1; or that is capable of providing a sequence polynucleotide SEQ ID NO: 3; or that is capable of providing a polynucleotide of sequence SEQ ID NO: 5.
  • said precursor is a nucleotide sequence that increases the bioavailability of the polynucleotides that it provides when administered to an individual or that enhances their release in a compartment biological.
  • the precursor nucleotide sequences of the invention are SEQ ID NO: 2, a sequence that will also be referred to as pre-miR-219a and that is a precursor to SEQ ID NO: 1; SEQ ID NO: 4, precursor sequence of SEQ ID NO: 3; and SEQ ID NO: 6, precursor sequence of SEQ ID NO: 5. All of these precursor nucleotide sequences comprise polynucleotides SEQ ID NO: 1, SEQ ID NO: 3 and SEQ ID NO: 5, respectively.
  • the medicaments and pharmaceutical compositions of the invention are useful for the treatment of arrhythmogenic canalopathies, preferably, for the treatment of arrhythmogenic canalopathies that are due to alterations in the sodium channels, since the polynucleotide of the invention is capable of reducing the expression of the SCN5A gene that codes for a transmembrane protein of sodium channels, so it can be used in those arrhythmogenic canalopathies due to alterations in function gain in these channels.
  • the polynucleotide of the invention comprising the nucleotide sequence SEQ ID NO: 5 or that of SEQ ID NO: 6, has a lower complementarity with the mRNA of this gene than the native microRNA, so competing with the latter for said Binding is able to increase the levels of mRNAs of this gene, compared to the native microRNA, so it is useful for the treatment of arrhythmogenic canalopathies due to impaired loss of function in the sodium channels.
  • another aspect of the invention relates to the use of the polynucleotide of the invention for the preparation of a medicament for the treatment of arrhythmogenic canalopathies, or alternatively, to the polynucleotide of the invention for use as a medicament for the treatment of arrhythmogenic canalopathies.
  • arrhythmogenic canalopathies are due to alterations in the sodium channels.
  • the arrhythmogenic canalopathy that is due to alterations in the sodium channels is the long QT syndrome type 3.
  • Another aspect of the invention relates to the use of the polynucleotide of the invention comprising the nucleotide sequence SEQ ID.
  • medication refers to any substance used for the prevention, diagnosis, relief, treatment or cure of diseases in man and women. animals. In the context of the present invention it refers to a preparation comprising at least the polynucleotide of the invention.
  • the polynucleotide of the invention is formulated in an appropriate pharmaceutical composition, in the therapeutically effective amount, preferably together with one or more pharmaceutically acceptable carriers, adjuvants or excipients.
  • treatment is to combat the effects caused as a consequence of arrhythmogenic canalopathies, preferably, of arrhythmogenic canalopathies that are due to alterations in the sodium channels, more preferably of the QT syndrome long type 3 or Brugada syndrome, to stabilize the condition of individuals or prevent further damage.
  • prevention is to prevent the occurrence of damage whose cause are arrhythmogenic canalopathies, preferably, arrhythmogenic canalopathies that are due to alterations in the sodium channels, more preferably the syndrome of Long QT type 3 or Brugada syndrome.
  • Arrhythmogenic canalopathy is understood as any genetic cardiomyopathy (or impaired myocardial function) that results from mutations in the genes responsible for the proper functioning of the ion channels that generate the potential for transmembrane action, which leads to defects in function ( gain or loss) of said channels, physiological alterations of the duration of the potential for transmembrane action and / or predisposition to develop ventricular arrhythmias in the absence of structural heart disease.
  • arrhythmogenic canalopathies include, but are not limited to, Brugada syndrome, long QT syndrome, short QT syndrome or catecholaminergic polymorphic ventricular tachycardia.
  • Arrhythmogenic canalopathies can be diagnosed, for example, but not limited to, as described in Cabrera Ortega M., et al., 2009, Revista Cubana Pediatr ⁇ a, 81 (4).
  • "Arrhythmogenic canalopathies due to alterations in sodium channels” are those canalopathies whose genetic basis is found in mutations (loss or gain of function) in at least one of the genes that code for said channels, preferably, in the SCN5A gene of SEQ ID NO: 7.
  • the detection of this type of canalopathies can be performed by, for example, but not limited to, an electrocardiogram, by means of which it is possible to make a potential diagnosis of the altered currents (Antzelevitch et al.
  • Brugada syndrome is characterized by the probability of presenting syncopal episodes or cardiac arrest caused by polymorphic ventricular tachycardia or ventricular fibrillation during rest or sleep, with an electrocardiographic pattern of apparent right bundle branch block and supra-level ST segment that falls slowly and ends on a negative T wave in V1, V2 and V3, without depression of the opposite leads. Genetically they have identified seven types of Brugada syndrome, the most frequent are, but not limited to, Brugada syndrome type 1 or Brugada syndrome type 2.
  • first pharmaceutical composition of the invention which comprises the polynucleotide of the invention comprising a nucleotide sequence with at least 60%, preferably with at least 65%, more preferably with at least 70%, with at least 75% , with at least 80%, with at least 85%, with at least 90%, with at least 95% or with at least 98% identity with respect to the full length of the sequence SEQ ID NO: 1, or comprising the nucleotide sequence SEQ ID NO: 1, or of SEQ ID NO: 2, or comprising the nucleotide sequence SEQ ID NO: 3 or of SEQ ID NO: 4.
  • the first pharmaceutical composition of the invention further comprises another active ingredient.
  • the first pharmaceutical composition of the invention further comprises a pharmaceutically acceptable carrier.
  • second pharmaceutical composition of the invention comprising the polynucleotide of the invention comprising the nucleotide sequence SEQ ID NO: 5 or SEQ ID NO: 6.
  • the second pharmaceutical composition of the invention further comprises another active ingredient.
  • the second pharmaceutical composition of the invention further comprises a pharmaceutically acceptable carrier.
  • active substance As used herein, the term “active substance”, “active substance”, “pharmaceutically active substance”, “active ingredient” or “ingredient pharmaceutically active” “means any component that potentially provides a pharmacological activity or other effect different in the diagnosis, cure, mitigation, treatment, or prevention of a disease, or that affects the structure or function of the body of man or other animals.
  • Adjuvants and “pharmaceutically acceptable carriers” refer to those substances, or combination of substances, known in the pharmaceutical sector, used in the preparation of pharmaceutical forms of administration and include, but are not limited to, solids, liquids, solvents or surfactants. .
  • Pharmaceutically acceptable carriers that can be used in the present invention are vehicles known in the state of the art, such as, but not limited to, lipid residues.
  • compositions and medicaments of the present invention can be used in a method of treatment or prevention in isolation or in conjunction with other pharmaceutical compounds intended for the treatment or prevention of arrhythmogenic canalopathies, preferably, of arrhythmogenic canalopathies that are due to alterations in the sodium channels, more preferably, of type 3 long QT syndrome or Brugada syndrome.
  • the pharmaceutical compositions of the present invention can be formulated for administration in a variety of ways known in the state of the art.
  • compositions and / or their formulations can be administered to an animal, including a mammal and, therefore, to man, in a variety of ways, including, but not limited to, parenteral, intraperitoneal, intravenous, intradermal, intraarticular, intrasynovial, intralesional, intraarterial, intracardiac, intramuscular, intranasal, subcutaneous, intracapsular, topical, by transdermal patches, by percutaneous administration, surgical implant, internal surgical painting, infusion pump or catheter.
  • the dosage to obtain a therapeutically effective amount depends on a variety of factors, such as the age, weight, sex or tolerance of the individual.
  • the term "therapeutically effective amount” refers to the amount of the pharmaceutical compositions of the invention that produces the desired effect and, in general, will be determined, among other causes, by the characteristics of said pharmaceutical compositions and the therapeutic effect to be achieved.
  • the medicament is for the treatment of arrhythmogenic canalopathies.
  • arrhythmogenic canalopathies are due to alterations in the sodium channels.
  • arrhythmogenic canalopathy due to alterations in sodium channels is the long QT syndrome type 3.
  • Another aspect of the invention relates to the use of the second pharmaceutical composition of the invention for the preparation of a medicament for the treatment of Brugada syndrome, or alternatively, to the second pharmaceutical composition of the invention for use as a medicament for the treatment. of Brugada syndrome.
  • Fig. 1 Shows the complementarity scheme of hsa-mir-219-5p (SEQ ID NO: 1) with the 3 UTR region of the SCN5A gene messenger.
  • Upper sequence 3 ' UTR region of the messenger of the SCN5A gene (position 1 .416-1 .422, 5 X - 3 " ).
  • Lower sequence sequence of hsa-mir-219-5p (SEQ ID NO: 1, 3 " - 5 " ) with the sequence complementary to the messenger of the SCN5A gene (GAUUGUC) marked in white.
  • Black bars complementary nucleotides.
  • Fig. 2. Shows the sequence of the mature hsa-mir-219-5p and its precursor.
  • Fig. 4 Shows the results of immunohistochemical tests against the expression product of the SCN5A gene in control cells (panel left) and in cells that were transfected with hsa-mir-219-5p (right panel).
  • the left panel shows the normal protein expression of Nav1 .5 (SCN5A) in the cardiomyocytic cell.
  • Nav1 .5 appears to be sequestered in the Golgi apparatus, and therefore is very poorly represented in the cytoplasmic membrane, where it normally exerts its sodium ion transport function.
  • Fig. 5 It shows an illustrative scheme of the strategy for the treatment of long QT syndrome type 3 where there is gain of function of the SCN5A gene.
  • A. Upper sequence: 3 ' UTR region of the messenger of the SCN5A gene (position 1 .416-1 .422, 5 " - 3 " ).
  • the box in the lower sequence shows the three nucleotides that have been replaced by the nucleotides shown in the lower part to give rise to SEQ ID NO: 3. The complementarity between both upper and lower sequences (bars) is shown.
  • the gray bars represent the new complementary sites created to give rise to SEQ ID NO: 3.
  • B Shows the precursor sequence of SEQ ID NO: 1, pre-miR-219a, of SEQ ID NO: 2, where the mature sequence of hsa-mir-219-5p (SEQ ID NO: 1) stands out in gray in the two complementary strands of the pre-miR-219a, in a box the three modified nucleotides are indicated to design SEQ ID NO: 3
  • C Shows SEQ ID NO: 3 (in bold) included in its precursor sequence or SEQ ID NO: 4. In gray, the modified nucleotides are highlighted.
  • Fig. 6. It shows an illustrative scheme of the strategy for the treatment of Brugada syndrome where there is loss of function of the SCN5A gene. TO.
  • Upper sequence 3 ' UTR region of the messenger of the SCN5A gene (position 1 .416-1 .422, 5 N - 3 " ).
  • Lower sequence sequence of hsa-mir-219-5p (SEQ ID NO: 1, 3 ' - 5 ' ) with the sequence complementary to the messenger of the SCN5A gene (GAUUGUC) marked in white.
  • the boxes in the lower sequence show the three nucleotides that have been replaced by the nucleotides that are shown at the bottom to give rise to SEQ ID NO: 5. The complementarity between both upper and lower sequences (bars) is shown.
  • FIG. 7 Shows the expression of the SCN5A gene in atrial myocardial cells (HL-1) transfected with SEQ ID NO: 2, SEQ ID NO: 4 or SEQ ID NO: 6 for 12 hours.
  • Control cells where the precursor of endogenous hsa-mir-219-5p or miR-219a (SEQ ID NO: 2) was transfected.
  • miR-219a- Cells transfected with the precursor of miR-219a- (SEQ ID NO: 4), designed to treat arrhythmogenic canalopathies due to a gain in function in the SCN5A gene.
  • miR-219a + Cells transfected with the precursor of miR-219 + (SEQ ID NO: 6), designed to treat arrhythmogenic canalopathies due to a loss of function in the SCN5A gene.
  • the figure shows that HL-1 cells transfected with SEQ ID NO: 4 show a decrease in SCN5A expression compared to cells transfected with the endogenous precursor of miR-219a (SEQ ID NO: 2), while those Myocardial cells transfected with SEQ ID NO: 6 show increased expression of SCN5A compared to cells transfected with the endogenous precursor of miR-219a (SEQ ID NO: 2).
  • EXAMPLE 1 Effect of the expression of hsa-mir-219-5p on the expression of the SCN5A gene in atrial myocardial cells.
  • the hsa-mir-219-5p (SEQ ID NO: 1) is capable of binding to the 3 ' UTR region of the mRNA of the SCN5A gene as shown in Figure 1, in particular by the region corresponding to the GAUUGUC sequence included in SEQ ID NO: 1 (marked in bold in Figure 2A).
  • This hsa-mir-219-5p is included in the sequence of its precursor or pre-miR-219a (SEQ ID NO: 2) as seen in Figures 2B and 2C.
  • hsa-mir-219-5p conditioned the contractile capacity of cardiomyocytes in culture.
  • contractions of control cardiomyocytes and cardiomyocytes transfected with hsa-mir-219-5p were counted, and it was observed that the rate was decreased by approximately 30% in the latter, and that the contraction pattern was asynchronous (Table 1). Therefore, these data revealed that hsa-mir-219-5p can regulate the function of the sodium channel, which is very important since it implies a molecular mechanism of easy manipulation and high accessibility that allows to correct the lack or gain of function of the Cardiac sodium channel underlying arrhythmogenic syndromes such as Brugada or long QT, respectively.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biophysics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Microbiology (AREA)
  • Cardiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne l'utilisation de micro-ARN hsa-mir-219-5ρ, de séquence SEQ ID NO: 1, des polynucléotides modifiés dérivés de celui-ci, SEQ ID NO: 3 ou SEQ ID NO: 5, ou de ses précurseurs, SEQ ID NO: 2, SEQ ID NO: 4 ou SEQ ID NO: 6, pour la préparation de médicaments utilisés pour le traitement de canalopathies arythmogènes, de préférence celles dues à des altérations du fonctionnement du canal sodique cardiaque, plus préférablement du syndrome du QT long de type 3 et du syndrome de Brugada. Ces polynucléotides sont capables de moduler le fonctionnement du canal sodique cardiaque par liaison à l'ARNm du gène SCN5A.
PCT/ES2011/000248 2010-08-02 2011-07-29 Micro-arn utilisé pour le traitement de canalopathies arythmogènes Ceased WO2012017105A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ESP201031205 2010-08-02
ES201031205A ES2374244B1 (es) 2010-08-02 2010-08-02 MicroRNA útil para el tratamiento de canalopatías arritmogénicas.

Publications (1)

Publication Number Publication Date
WO2012017105A1 true WO2012017105A1 (fr) 2012-02-09

Family

ID=45531646

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ES2011/000248 Ceased WO2012017105A1 (fr) 2010-08-02 2011-07-29 Micro-arn utilisé pour le traitement de canalopathies arythmogènes

Country Status (2)

Country Link
ES (1) ES2374244B1 (fr)
WO (1) WO2012017105A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105385688A (zh) * 2015-11-05 2016-03-09 宁波市医疗中心李惠利医院 一种与长QT综合征相关的miRNA及其应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100010073A1 (en) * 2006-10-09 2010-01-14 Julius-Maximilians-Universität Würzburg Microrna (mirna) for the diagnosis and treatment of heart diseases

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100010073A1 (en) * 2006-10-09 2010-01-14 Julius-Maximilians-Universität Würzburg Microrna (mirna) for the diagnosis and treatment of heart diseases

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BLANCHE SCHOROEN ET AL.: "MicroRNAs and beyond: the heart reveals its treasures", HYPERTENSION, vol. 54, 19 October 2009 (2009-10-19), pages 1189 - 1194, XP002600450, DOI: doi:10.1161/HYPTERTENSIONAHA.109.133942 *
HEATHER S. DUFFY: "The ever-shrinking world of cardiac ion channel remodeling: the role of microRNAs in heart disease", HEART RHYTHM SOCIETY, vol. 6, no. 12, 2009, pages 1810 - 1811, XP026857385 *
STANLEY ET AL.: "Ion-channel mRNA-expression profiling: insights into cardiac remodeling and arrhythmic substrates", JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, vol. 48, 23 July 2009 (2009-07-23), pages 96 - 105 *
XIAOBIN LUO ET AL.: "Regulation of human cardiac ion channel genes by microRNAs: theoretical perspective and pathophysiological implications", CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, vol. 25, no. 6, 18 May 2010 (2010-05-18), pages 571 - 586 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105385688A (zh) * 2015-11-05 2016-03-09 宁波市医疗中心李惠利医院 一种与长QT综合征相关的miRNA及其应用

Also Published As

Publication number Publication date
ES2374244B1 (es) 2012-12-26
ES2374244A1 (es) 2012-02-15

Similar Documents

Publication Publication Date Title
US8916532B2 (en) Methods for enhancing utrophin production via inhibition of microRNA
JP2021100435A (ja) 調節性ポリヌクレオチド
CN115867657A (zh) 用于治疗疾患和疾病的opa1反义寡聚物
WO2017187426A1 (fr) Inhibition de l'arnmi mir-22 par apt-110
JP2022169627A5 (fr)
Grisorio et al. Gene therapy for cardiac diseases: methods, challenges, and future directions
JP2019523302A5 (fr)
Kim et al. The role and medical prospects of long non-coding RNAs in cardiovascular disease
JP2011505798A5 (fr)
ES2374244B1 (es) MicroRNA útil para el tratamiento de canalopatías arritmogénicas.
CN102416184A (zh) microRNA-1的反义锁核苷酸序列在制备预防或治疗心肌梗死后心衰药物中的应用
US20160108431A1 (en) Prevention and treatment of atrial fibrillation
US10357478B2 (en) Polyamide compound and pharmaceutical composition for treating mitochondrial genetic diseases
CN113811313A (zh) 食管狭窄抑制剂
CN113755496B (zh) 小rna及其在糖尿病治疗中的用途
JP2024511207A (ja) ハンチントン病を治療するためのｒｎａ送達システム
CN120899738A (zh) 一种miR-411变体在制备预防和/或治疗心梗后心肌纤维化的药物中的应用
CN101642462B (zh) 一种肿瘤抑制剂nrn1sr42
Zhong et al. Dilated Cardiomyopathy and Genotype-Targeted Therapeutic
CN119185545A (zh) Dot1l作为阿片类药物诱发痛觉过敏的治疗靶点的应用
WO2024112653A1 (fr) Acides nucléiques inhibiteurs et leurs procédés d'utilisation
CN101503437B (zh) 核酸分子si-cypj-4及其在制备抗癌药物中的应用
CN116801916A (zh) 包含miR 145抑制剂作为有效成分的用于治疗心肌梗塞的组合物
CN101642463A (zh) 一种肿瘤抑制剂nrn1sr4
JPWO2022221777A5 (fr)

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11814137

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11814137

Country of ref document: EP

Kind code of ref document: A1