WO2021080396A1 - Composition pour la prévention ou le traitement d'une cardiopathie valvulaire comprenant un inhibiteur de rspo3 - Google Patents

Composition pour la prévention ou le traitement d'une cardiopathie valvulaire comprenant un inhibiteur de rspo3 Download PDF

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WO2021080396A1
WO2021080396A1 PCT/KR2020/014647 KR2020014647W WO2021080396A1 WO 2021080396 A1 WO2021080396 A1 WO 2021080396A1 KR 2020014647 W KR2020014647 W KR 2020014647W WO 2021080396 A1 WO2021080396 A1 WO 2021080396A1
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rspo3
valve
expression
calcification
valve calcification
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PCT/KR2020/014647
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English (en)
Korean (ko)
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장기육
박은혜
김은민
강권윤
김찬우
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가톨릭대학교 산학협력단
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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
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • 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

Definitions

  • the present invention relates to a composition for preventing or treating heart valve disease comprising an inhibitor of RSPO3 expression or activity as an active ingredient.
  • Valvular heart disease is a disease in which the opening and closing of the valve is unfavorable.
  • a case in which the valve does not open well functionally is called stenosis. It opens well but does not close tightly, so blood flows back. The case is called insufficiency/regurgitation. Since there are 4 rooms in the human heart, there are 4 valves, and most of the clinically problematic areas are the aortic valve and 2 mitral valves.
  • Calcific aortic valve disease is a disease caused by calcification of the aortic valve, and is one of the most common heart valve diseases, and the incidence of the disease is gradually increasing as the elderly population increases. . Calcific aortic valve disease can progress to aortic valve sclerosis and then to aortic valve stenosis.
  • valve replacement surgery in which the diseased valve is removed and replaced with an artificial valve
  • plastic surgery has recently been introduced to repair the valve and continue to use it.
  • it is necessary to develop a material capable of inhibiting calcification of the aortic valve in that it is necessary to consider the risks of the operation itself and the problems that may occur after the operation.
  • An object of the present invention is to provide a pharmaceutical composition for preventing or treating vascular or valve calcification comprising an inhibitor of expression or activity of RSPO3 (R-spondin 3) as an active ingredient.
  • Another object of the present invention is the steps of: 1) administering a candidate substance to an individual in which blood vessel or valve calcification is induced;
  • step 3 the step of confirming the decrease in the expression of the RSPO3 gene or protein compared to the control; to provide a screening method for a candidate material for the prevention or treatment of blood vessel or valve calcification comprising.
  • Another object of the present invention is to provide a method for preventing and treating blood vessels or calcification, comprising administering to a subject a pharmaceutically effective amount of the pharmaceutical composition.
  • the present invention provides a pharmaceutical composition for preventing or treating vascular or valve calcification comprising an inhibitor of expression or activity of RSPO3 (R-spondin 3) as an active ingredient.
  • RSPO3 R-spondin 3
  • the present invention comprises the steps of: 1) administering a candidate substance to an individual in which blood vessel or valve calcification is induced;
  • step 3 the step of confirming the decrease in the expression of the RSPO3 gene or protein compared to the control; it provides a screening method for a candidate material for the prevention or treatment of blood vessel or valve calcification comprising.
  • the present invention provides a method for preventing and treating blood vessels or calcification, comprising administering to an individual a pharmaceutically effective amount of the pharmaceutical composition.
  • the present invention relates to a composition for preventing or treating valvular heart disease comprising an inhibitor of expression or activity of RSPO3 as an active ingredient, wherein the inhibitor of expression or activity of RSPO3 can inhibit calcification of blood vessels or valves. Yes, it can be usefully used in the prevention or treatment of related diseases.
  • control group VIC cells derived from valve (CAVD) with calcified lesion formation were not treated with Pro-calcific medium (patient(-) group)); Or it shows the RNA sequencing analysis method in the treatment group (patient (+) group).
  • FIG. 1B is a group of normal valve-derived VIC cells (control, control); VIC cells derived from valve (CAVD) with calcified lesion formation were not treated with Pro-calcific medium (patient(-) group)); Alternatively, the results of RNA sequencing analysis in the treatment group (patient (+) group) are shown.
  • 2A is a group of normal valve-derived VIC cells; And a gene ontology (GO) analysis in the group treated with Pro-calcific medium to valve-derived VIC cells with calcified lesion formation and upregulated genes.
  • GO gene ontology
  • 2B is a group of normal valve-derived VIC cells; And a gene ontology (GO) analysis in the group treated with Pro-calcific medium to valve-derived VIC cells with calcified lesion formation and down-regulated genes.
  • GO gene ontology
  • Figure 3A shows the results of measuring the expression level of BMP2 through real-time PCR after each treatment with Pro-calcific medium on normal valve-derived VIC cells and valve-derived VIC cells with calcified lesion formation (AR(- ): aortic regurgitation(control) without pro-calcific medium; AR(+): aortic regurgitation(control) with pro-calcific medium; AS(-): aortic stenosis(patient) without pro-calcific medium; and AS(+) : aortic stenosis(patient) with pro-calcific medium).
  • AR(- ) aortic regurgitation(control) without pro-calcific medium
  • AR(+) aortic regurgitation(control) with pro-calcific medium
  • AS(-) aortic stenosis(patient) without pro-calcific medium
  • AS(+) aortic stenosis(patient) with pro-calcific medium
  • 3B shows the results of measuring the expression level of RUNX2 through real-time PCR after each treatment with Pro-calcific medium on normal valve-derived VIC cells and valve-derived VIC cells with calcified lesion formation
  • AR(- ) aortic regurgitation(control) without pro-calcific medium
  • AR(+) aortic regurgitation(control) with pro-calcific medium
  • AS(-) aortic stenosis(patient) without pro-calcific medium
  • AS(+) aortic stenosis(patient) with pro-calcific medium
  • Figure 3C shows the results of measuring the expression level of RSPO3 through real-time PCR after each treatment with Pro-calcific medium on normal valve-derived VIC cells and valve-derived VIC cells with calcified lesion formation
  • AR(- ) aortic regurgitation(control) without pro-calcific medium
  • AR(+) aortic regurgitation(control) with pro-calcific medium
  • AS(-) aortic stenosis(patient) without pro-calcific medium
  • AS(+) aortic stenosis(patient) with pro-calcific medium
  • 4A shows the results of performing Alizarin Red S staining and immunohistochemistry for RSPO3 on normal valve tissue (Non-CAVD).
  • 4B shows the results of performing Alizarin Red S staining and immunohistochemistry for RSPO3 on valve tissue (CAVD) of a patient with calcific aortic valve disease.
  • 5A shows the results of immunohistochemistry for RSPO3 in normal valve tissue (Non-CAVD).
  • 5B shows the results of immunohistochemistry for RSPO3 in valve tissue (CAVD) of a patient with calcific aortic valve disease.
  • 6A is a group treated with control siRNA in valve-derived VIC cells with calcified lesion formation (siControl Cal-); Valve-derived VIC cells with calcified lesion formation were treated with Pro-calcific medium and control siRNA (siControl Cal+); And it shows the result of measuring the expression level of BMP2 through real-time PCR for the group (siRSPO3 Cal+) treated with Pro-calcific medium and treated with RSPO3 siRNA in VIC cells derived from valves with calcified lesion formation.
  • 6B is a group (siControl Cal-) treated with control siRNA in valve-derived VIC cells with calcified lesion formation; Valve-derived VIC cells with calcified lesion formation were treated with Pro-calcific medium and control siRNA (siControl Cal+); And it shows the result of measuring the expression level of RSPO3 through real-time PCR for the group (siRSPO3 Cal+) treated with Pro-calcific medium and the RSPO3 siRNA-treated group (siRSPO3 Cal+) in valve-derived VIC cells with calcified lesion formation.
  • 6C is a group (siControl) treated with Pro-calcific medium and control siRNA in valve-derived VIC cells with calcified lesion formation; And it shows the results of performing Alizarin Red S staining on the group (siRSPO3) treated with Pro-calcific medium and treated with RSPO3 siRNA on VIC cells derived from valves with calcified lesion formation.
  • the present invention provides a pharmaceutical composition for preventing or treating vascular or valve calcification comprising an inhibitor of expression or activity of RSPO3 (R-spondin 3) as an active ingredient.
  • RSPO3 R-spondin 3
  • vascular or valve calcification refers to the formation, growth or deposition of extracellular matrix hydroxyapatite (calcium phosphate) crystal deposits in blood vessels or valves.
  • the vascular calcification includes calcification of coronary arteries, aorta and other blood vessels, and may be, for example, medialcalcification or atherosclerotic calcification.
  • the mesenteric calcification can be used in the same meaning as medial wall calcification or Moenckeberg's sclerosis, and means calcification by calcium deposited in the medial wall. It is known that the mesenteric calcification is caused by the expression of bone formation promoting factors in blood vessels due to diabetes, abnormal calcium metabolism, or kidney disease.
  • the atherosclerotic calcification refers to calcification occurring in artheromatous plaques along the inner membrane layer. It is known that such atherosclerotic calcification is caused by depositing calcium directly at the site of atherosclerotic lesions.
  • the blood vessel or valve calcification may be induced in human heart, lung, kidney and aorta, and is not limited to all tissues and organs in which calcification may occur.
  • heart valve disease is caused, which is a disease in which the opening and closing action of the valve is unfavorable, and a case in which the valve is not functionally opened is called stenosis, but it is well opened.
  • stenosis a case in which the valve is not functionally opened
  • insufficiency/regurgitation When the blood flows back because it is not tightly closed, it is called insufficiency/regurgitation.
  • the heart valve disease includes calcific aortic valve disease (CAVD), aortic stenosis, mitral stenosis, aortic regurgitation, and mitral valve insufficiency.
  • CAVD calcific aortic valve disease
  • aortic stenosis aortic stenosis
  • mitral stenosis mitral stenosis
  • mitral valve insufficiency may be any one selected from the group consisting of, and preferably, calcific aortic valve disease or aortic valve stenosis, but is not limited thereto.
  • the calcification aortic valve disease or aortic valve stenosis may be caused by calcification of the aortic valve.
  • the RSPO3 may be composed of the amino acid sequence of SEQ ID NO: 1, and the gene encoding RSPO3 may be composed of the nucleotide sequence of SEQ ID NO: 2.
  • the RSPO3 expression or activity inhibitor may be a RSPO3 gene expression inhibitor or an RSPO3 protein expression or activity inhibitor.
  • the RSPO3 gene expression inhibitor is selected from the group consisting of antisense oligonucleotide, siRNA (small interfering RNA), shRNA (short hairpin RNA), and aptamer that bind complementarily to the mRNA of the RSPO3 gene. It may be any one, but is not limited thereto.
  • the RSPO3 protein expression or activity inhibitor is any selected from the group consisting of compounds, peptides, peptide mimetics, substrate analogs, aptamers, and antibodies that specifically bind to RSPO3 protein. It may be one, but is not limited thereto.
  • the "antisense oligonucleotide” is a DNA, RNA, or derivative thereof containing a nucleic acid sequence that is complementary to the mRNA nucleotide sequence of the target gene, and binds to the complementary sequence in the mRNA to inhibit the translation of the mRNA into a protein. do.
  • Double consisting of a 15 to 30 bp long sense sequence and an antisense sequence capable of inhibiting the expression of the target gene through RNAi (RNA interference) by mRNA cleavage by complementarily binding to the mRNA nucleotide sequence of the "siRNA" target gene As -stranded RNA, it can be used for gene knockdown or gene therapy.
  • RNAi RNA interference
  • the siRNA is an RNA molecule of the sense sequence (UGUGAUACCUGUUUCAACAtt) consisting of the nucleotide sequence of SEQ ID NO: 3 and the antisense sequence (UGUUGAAACAGGUAUCACAgt) consisting of the nucleotide sequence of SEQ ID NO: 4 capable of complementarily binding to the mRNA of the RSPO3 gene.
  • the sense sequence UGUGAUACCUGUUUCAACAtt
  • UGUUGAAACAGGUAUCACAgt the antisense sequence consisting of the nucleotide sequence of SEQ ID NO: 4 capable of complementarily binding to the mRNA of the RSPO3 gene.
  • the “shRNA” is an RNA having a hairpin structure capable of silencing the expression of the target gene through RNAi (RNA interference) by complementary binding to the mRNA nucleotide sequence of the target gene. After being cleaved into siRNA, it binds to an RNA-induced silencing complex (RISC), and RISC binds to and cleaves mRNA, which can be used to inhibit gene expression.
  • RISC RNA-induced silencing complex
  • peptide mimetic refers to a peptide or non-peptide that inhibits the activity of the protein of interest by inhibiting the binding domain of the protein of interest.
  • the "aptamer” is an oligonucleotide or peptide molecule that specifically binds to a target molecule, and in the case of a nucleic acid aptamer, it is an evolutionary method using an oligonucleotide library called SELEX (systematic evolution of ligands by exponential enrichment). Thus, it can be obtained by separating oligomers that bind to specific chemical molecules or biological molecules with high affinity and selectivity.
  • the aptamer can specifically bind to a target and modulate the activity of the target, such as blocking the function of the target through binding.
  • the "antibody” refers to a substance that reacts when an external substance, which is an antigen, invades while circulating in blood or lymph in the immune system of a living body, and is a globulin-based protein formed in lymphatic tissues and is also called immunoglobulin. Antibodies are produced by B cells and specifically bind to antigens.
  • One antibody molecule has two heavy chains and two light chains, and each heavy chain and light chain each contain a variable region at the N-terminus.
  • Each variable region consists of three complementarity determining regions (CDRs) and four framework regions (FRs), and the complementarity determining regions determine the antigen binding specificity of the antibody, and the structure of the variable region. It exists as a relatively short peptide sequence that is maintained at the forming sites.
  • Such antibodies are polyclonal antibodies, monoclonal antibodies, recombinant antibodies and in complete form having two full-length light chains and two full-length heavy chains, as well as functional fragments of antibody molecules, e.g., Fab, F(ab' ), F(ab')2 and Fv.
  • the pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable means exhibiting properties that are not toxic to cells or humans exposed to the composition.
  • the carrier may be used without limitation as long as it is known in the art such as a buffering agent, a preservative, a painless agent, a solubilizing agent, an isotonic agent, a stabilizer, a base agent, an excipient, and a lubricant.
  • the pharmaceutical composition of the present invention is formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions according to a conventional method.
  • oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc.
  • external preparations suppositories
  • suppositories sterile injectable solutions
  • sterile injectable solutions according to a conventional method.
  • I can.
  • it may be used in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel or gel for external skin.
  • Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oils.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient, such as starch, calcium carbonate, and sucrose ( sucrose), lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
  • Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.
  • Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories.
  • non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.
  • injectable ester such as ethyl oleate
  • a base for suppositories witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • administration means introducing a predetermined substance to an individual by an appropriate method, and the route of administration of the composition may be administered through any general route as long as it can reach the target tissue.
  • Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration may be administered, but are not limited thereto.
  • the term "individual” refers to all animals including humans, such as rats, mice, and domestic animals. Preferably, it may be a mammal including a human.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not cause side effects, and the effective dose level is the sex, age, and Weight, health condition, type of disease, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration, and rate of excretion, duration of treatment, factors including drugs used in combination or concurrently, and other fields of medicine. It can be readily determined by a person skilled in the art according to well-known factors. Administration may be administered once a day at the recommended dosage, or may be divided several times.
  • the present invention comprises the steps of: 1) administering a candidate substance to an individual in which blood vessel or valve calcification is induced;
  • step 3 the step of confirming the decrease in the expression of the RSPO3 gene or protein compared to the control; it provides a screening method for a candidate material for the prevention or treatment of blood vessel or valve calcification comprising.
  • the method may further include the step of selecting a candidate substance whose expression level of the RSPO3 protein or a gene encoding the same is reduced from a reference value.
  • the measurement was RT-PCR (reverse transcription polymerase chain reaction), competitive RT-PCR, real-time quantitative RT-PCR, RNase protection method, northern blot, DNA chip technology. assay), ELISA (Enzyme-linked immunosorbent assay), Western blot, immunoblot, or immunocytochemistry.
  • the present invention provides a composition for diagnosis of valvular heart disease comprising an agent capable of measuring the expression level of RSPO3 (R-spondin 3) protein or a gene encoding the same.
  • RSPO3 R-spondin 3
  • the agent for measuring the expression level of the protein may be an antibody, an interacting protein, a ligand, nanoparticles, or aptamer that specifically binds to the protein or peptide fragment, but is not limited thereto. .
  • the agent for measuring the expression level of the gene encoding the protein may be a primer or a probe that binds to the mRNA of the gene, but is not limited thereto.
  • the term "primer” is a base sequence having a short free 3-terminal hydroxyl group, which can form a base pair with a complementary template, and as a starting point for template strand copying It refers to a short sequence of action.
  • Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerate or reverse transcriptase) and four different nucleoside triphosphates at an appropriate buffer and temperature. PCR conditions, the length of the sense and antisense primers can be appropriately selected according to techniques known in the art.
  • the term "probe” refers to a nucleic acid fragment such as RNA or DNA capable of specifically binding to a gene, and is labeled so that the presence or absence of a specific gene and the amount of expression can be confirmed.
  • the probe may be manufactured in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. Selection of an appropriate probe and conditions for hybridization can be appropriately selected according to techniques known in the art.
  • the primer or probe of the present invention can be chemically synthesized using a method for synthesizing a phosphoramidite solid support or other well-known methods.
  • These nucleotide sequences can also be modified through various methods known in the art. Examples of such modifications include methylation, encapsulation, substitution of one or more homologs of natural nucleotides, and modifications between nucleotides, such as uncharged linkers (e.g. methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.). ) Or to a charged linker (e.g. phosphorothioate, phosphorodithioate, etc.).
  • uncharged linkers e.g. methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.
  • a charged linker e.g. phosphorothioate, phosphorodithioate, etc.
  • the present invention provides a method for preventing and treating vascular or valve calcification, comprising administering to an individual a pharmaceutically effective amount of the pharmaceutical composition.
  • pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type and severity of the subject, age, sex, activity of the drug, and the drug. Sensitivity, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field.
  • the present inventors obtained normal valves without calcified lesion formation and valves with calcified lesion formation from patients who underwent aortic valve replacement, and after separating and culturing VIC (valvular interstitial cell) cells from the valve, respectively, through RNA sequencing analysis.
  • VIC valvular interstitial cell
  • the experiment was conducted in the normal valve-derived VIC cell group (control), the valve-derived VIC cells with calcified lesion formation without Pro-calcific medium (patient(-) group)) or treatment group (patient(+) group). Implemented (Fig. 1A). Each cell was incubated for 7 days and then used for analysis.
  • the present inventors confirmed that expression of various genes is regulated depending on whether or not calcified valve disease and whether calcification is induced.
  • the present inventors identified genes whose expression is increased in valve-derived VIC cells with calcified lesion formation compared to normal valve-derived VIC cells as a control group, and whose expression is further increased upon induction of calcification. It was determined to be, and the rate of increase in the expression of the gene was analyzed. As a result, RSPO3 (R-spondin 3) was upregulated by 3.16 fold in valve-derived VIC cells with calcified lesion formation compared to normal valve-derived VIC cells, and upregulated by 2.74 fold when inducing calcification, and the expression was most markedly increased. It was confirmed to be a gene.
  • the present inventors compared RSPO3 (R-spondin 3) with bone morphogenetic protein 2 (BMP2) and RUNX2 (Runt-related transcription factor 2), which are known as calcification-related genes, and the expression level of the gene in valve-derived VIC cells with calcified lesion formation. An experiment was conducted to verify whether it was high or not. Briefly, normal valve-derived VIC cells and valve-derived VIC cells with calcified lesion were treated with Pro-calcific medium and cultured for 4 and 7 days, respectively, and the expression level of the gene was confirmed through real-time PCR. .
  • the present inventors verified that the RSPO3 gene is upregulated in calcified valve-derived VIC cells.
  • the present inventors performed an experiment comparing the expression of RSPO3 according to the degree of calcification in human aortic valve tissue. Briefly, human aortic valve samples isolated from patients were fixed in 4% formalin solution, embedded in paraffin, and prepared as 4 ⁇ m-thick sections. Alizarin Red S staining was performed to confirm the degree of calcification of the valve tissue. As a result, it was confirmed that the valves of patients with aortic valve stenosis (limbic aortic valve disease) showed remarkable calcification compared to normal valves (FIGS. 4A and 4B).
  • RSPO3 aortic valve stenosis (limbic aortic valve disease) compared to normal valves (FIGS. 4A, 4B, 5A, and 5B).
  • the present inventors confirmed that the valves of patients with aortic valve stenosis (limbic aortic valve disease) showed remarkable calcification compared to normal valves, and that the expression of RSPO3 was remarkably high.
  • valve-derived VIC cells with calcified lesion formation were treated with control siRNA (siControl Cal-); Valve-derived VIC cells with calcified lesion formation were treated with Pro-calcific medium and control siRNA (siControl Cal+); And for the group (siRSPO3 Cal+) treated with Pro-calcific medium and treated with RSPO3 siRNA in valve-derived VIC cells with calcified lesion formation, the expression levels of BMP2 and RSPO3 were measured through real-time PCR.
  • the present inventors performed Alizarin Red S staining in order to compare the degree of calcification according to treatment with RSPO3 siRNA. As a result, it was confirmed that the treatment with RSPO3 siRNA (siRSPO3) had the effect of significantly reducing calcification compared to the treatment with the control siRNA (FIG. 6C).
  • the present inventors confirmed that the calcification of the aortic valve can be suppressed through the suppression of RSPO3 expression. Accordingly, the inventors of the present invention can be used for the prevention or treatment of heart valve diseases such as calcific aortic valve disease or aortic stenosis by inhibiting the calcification of the aortic valve by the inhibitor of RSPO3 expression or activity. Confirmed that there is.

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Abstract

La présente invention concerne une composition pour la prévention ou le traitement d'une cardiopathie valvulaire, comprenant un inhibiteur de l'expression ou de l'activité de RSPO3 en tant que principe actif. L'inhibiteur de l'expression ou de l'activité de RSPO3 peut inhiber la calcification des vaisseaux sanguins ou des valvules, et peut ainsi être utile dans la prévention ou le traitement des maladies associées.
PCT/KR2020/014647 2019-10-24 2020-10-26 Composition pour la prévention ou le traitement d'une cardiopathie valvulaire comprenant un inhibiteur de rspo3 WO2021080396A1 (fr)

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Cited By (1)

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Publication number Priority date Publication date Assignee Title
CN113667733A (zh) * 2021-07-12 2021-11-19 华中科技大学同济医学院附属协和医院 circRNA PRDM5在钙化性主动脉瓣疾病的诊断试剂盒及治疗药物开发中的应用

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