Arteriosclerosis severity diagnostic marker and diagnostic method using the same

Abstract

To provide a composition for preventing or treating arteriosclerosis.SOLUTION: The present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis, comprising an LGI 3 antagonist represented by a specific amino acid sequence as an active ingredient. Administering the LGI 3 antagonist can treat arteriosclerosis without affecting other metabolic phenotypes.SELECTED DRAWING: Figure 7

Description

[Technical field]

[0001] The present invention relates to a marker composition for diagnosing the severity of arteriosclerosis, a ... kit for diagnosing the severity of arteriosclerosis containing the composition, a method for screening a substance for preventing or treating the severity of arteriosclerosis, and a method for providing information regarding the diagnosis of the severity of arteriosclerosis.

[0002] This application claims priority based on Korean Patent Application No. 10-2018-0140320 filed on November 14, 2018, and Korean Patent Application No. 10-2019-0140180 filed on November 5, 2019, the entire contents of which are incorporated herein by reference in their entirety in their specification and drawings. [Background technology]

[0003] Atherosclerosis, also known as atherosclerosis or atherosclerosis, is a vascular disease in which cholesterol is deposited mainly in the endothelium, which covers the innermost layer of blood vessels, causing proliferation of endothelial cells, resulting in the formation of atheromas. It progresses gradually over a long period of time and is most prevalent in the coronary arteries, carotid arteries, lower abdominal aorta, and popliteal arteries, and depending on the site of onset, it can induce fatal diseases such as angina pectoris, cerebral infarction, cerebral hemorrhage, renal failure, ischemic limb disease, stroke, or myocardial infarction.

[0004] Depending on the severity of arteriosclerosis, blood vessels become narrowed and clogged, causing blood circulation problems in the areas served by those blood vessels, and symptoms appear due to the narrowed blood vessels. It is known that the more risk factors there are, the faster arteriosclerosis progresses, and the main risk factors include hypercholesterolemia, HDL cholesterol deficiency, LDL cholesterol excess, high triglycerides, high blood pressure, smoking, diabetes, family history of cardiovascular disease, increasing age, lack of exercise, excess weight and abdominal obesity. However, there is a drawback in that the severity of arteriosclerosis cannot be confirmed by checking only the main risk factors, as the main risk factors are not proportional to the severity of arteriosclerosis.

[0005] The severity of arteriosclerosis is an important factor in determining the treatment method (e.g., surgical procedure, etc.). Furthermore, the severity determines the major adverse cardiac and cerebrovascular event-free survival rate (MACCE) and increases the possibility of developing very dangerous clinical symptoms such as stroke or myocardial infarction. This makes it very important to diagnose the severity of arteriosclerosis. As part of the method of diagnosing the severity of arteriosclerosis, there is a method to confirm the degree of coronary artery stenosis. However, since the degree of coronary artery stenosis does not match the symptoms, the limitations of coronary angiography have been raised in assessing whether the stenosis is severe enough to cause symptoms, early detection and mechanical investigation of gradually progressing coronary artery disease, and reduction of coronary artery atherosclerosis by adjusting risk factors.

[0006] This has created an emerging need to develop methods to identify arteriosclerosis and, therefore, accurately diagnose the severity of arteriosclerosis.

[0007] On the other hand, LGI3 (leucine rich repeat LGI family member 3 or leucine rich glioma inactivated 3) is a secretory protein that exists only in vertebrates including humans, and is known to function in regulating secretion and differentiation in nerve cells, regulating survival and migration of skin keratinocytes, regulating pigment production in melanocytes, regulating differentiation of adipocytes and inflammation in obese adipose tissue. The LGI3 gene is located on human chromosome 8p21.3, has Gene ID: 203190, has 8 exons, and is composed of 548 amino acids (NCBI Reference Sequence: NP_644807, SEQ ID NO: 1). In addition, the LGI3 cDNA cloned from the mouse (Mus musculus) brain cDNA library is composed of 2931 nucleotides, and like humans, the mouse is composed of 548 amino acids (NCBI Reference Sequence: NP_660254.1, SEQ ID NO: 2).

[0008] However, based on research on LGI3 to date, it is not known at all whether the expression or activity level of LGI3 increases proportionally with the severity of arteriosclerosis. Summary of the Invention [Problem to be solved by the invention]

[0009] The problem that the present invention aims to solve is to provide a marker composition for diagnosing the severity of arteriosclerosis, a composition for diagnosing the severity of arteriosclerosis, a kit for diagnosing the severity of arteriosclerosis containing the composition, a method for screening substances for preventing or treating arteriosclerosis, and a method for providing information regarding the diagnosis of the severity of arteriosclerosis, which utilize the expression or activity level of LGI3.

[0010] However, the technical problems that the present invention aims to achieve are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those having ordinary skill in the art to which the present invention pertains from the following description. [Means for solving the problem]

[0011] In order to achieve the above objectives, The present invention provides a method for providing information for diagnosing the severity of arteriosclerosis, comprising the following steps: (a) determining the expression or activity level of LGI3 (leucine rich repeat LGI family member 3) represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a sample; and (b) a step of diagnosing the severity of arteriosclerosis as severe if the expression or activity level in step (a) is increased compared to the expression or activity level of LGI3 shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a control group.

[0012] In one embodiment of the present invention, the arteriosclerosis may be coronary artery disease or peripheral arterial disease.

[0013] According to an embodiment of the present invention, the control group may be a sample collected at a different time from the patient from whom the sample was collected in step (a).

[0014] In another embodiment of the present invention, The information providing method provides a method for diagnosing arteriosclerosis severity as well as major adverse cardiac and cerebrovascular event (MACCE) disease-free-survival rates, characterized in that:

[0015] Here, if the expression or activity level of LGI3 shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 is increased compared to the expression or activity level of LGI3 shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a control group, the patient is diagnosed as having a low disease-free survival rate for major cardiac and cerebrovascular diseases.

[0016] In another embodiment of the present invention, the major cardiac and cerebrovascular disease may be myocardial infarction or stroke.

[0017] In yet another embodiment of the present invention, the information providing method for diagnosing the severity of coronary artery disease may be performed in place of or in parallel with SYNTAX (the Synergy between PCI with Taxus and Cardiac Surgery).

[0018] In yet another embodiment of the present invention, for diagnosing the severity of peripheral arterial disease, the information providing method may replace or be performed in parallel with TASC (Trans-Atlantic Inter-Society Consensus).

[0019] The present invention also provides a biomarker composition for diagnosing the severity of atherosclerosis, comprising LGI3 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3.

[0020] The present invention also provides a biomarker composition for diagnosing the severity of arteriosclerosis, comprising a preparation for measuring the expression or activity level of LGI3 represented by the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3.

[0021] The present invention also provides a kit for diagnosing the severity of arteriosclerosis, comprising the composition.

[0022] The present invention also provides a method for screening a substance for preventing or treating arteriosclerosis, comprising the steps of: (a) confirming the expression or activity level of LGI3 (leucine rich repeat LGI family member 3) shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 after treatment with the candidate substance; and (b) selecting the candidate substance as a substance for preventing or treating arteriosclerosis if the expression or activity level in (a) is downward.

[0023] The present invention also provides an application for preventing and / or treating arteriosclerosis, which comprises an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO:3 as an active ingredient.

[0024] More specifically, the present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis, which contains as an active ingredient an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3. The present invention also provides a method for treating arteriosclerosis by administering an effective amount of an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 to an arteriosclerosis patient. The present invention also provides a use of the LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 for treating or preventing arteriosclerosis. Effect of the Invention

[0025] According to the present invention, the severity of arteriosclerosis can be accurately and easily diagnosed simply by confirming the expression or activity level of LGI3, which can be used as an important indicator for grasping the condition of an arteriosclerosis patient (whether or not there is improvement or recurrence, etc.) and determining the appropriateness of a current treatment method, thereby enabling the realization of personalized medicine and predictive medicine. In addition, according to the present invention, it is possible to develop a therapeutic substance for improving the severity of arteriosclerosis, and it is expected that the present invention will provide a stepping stone for using such a therapeutic substance in various fields such as pharmaceutical, topical, and food additive compositions. Furthermore, according to the present invention, it is expected that it is possible to treat arteriosclerosis without affecting metabolic phenotype, and to treat arteriosclerosis without side effects. [Brief description of the drawings]

[0026] [Figure 1] FIG. 1 shows that LGI3 deficiency reduces atherosclerotic plaque formation. [Diagram 2] FIG. 1 shows that LGI3 deficiency reduces aortic sinus pathology formation. [Diagram 3] FIG. 1 shows that LGI3 deficiency reduces body weight gain and inguinal white fat mass. [Figure 4] FIG. 1 shows that LGI3 is increased in plasma of an arteriosclerosis mouse model. [Diagram 5] FIG. 1 shows that LGI3 antagonist (LGI3 p34) reduces atherosclerotic plaque formation in Apoe− / − mice. [Figure 6] FIG. 1 shows that treatment with an LGI3 antagonist (LGI3 p34) does not alter the metabolic phenotype. [Figure 7] FIG. 1 shows the relationship between LGI3 expression levels and coronary artery disease (CAD). [Figure 8] FIG. 1 shows the relationship between LGI3 expression levels and peripheral artery disease (PAD). [Figure 9]FIG. 1 shows the relationship between LGI3 expression levels and risk factors for major adverse cardiac and cerebrovascular events (MACCE). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] In the following, preferred examples are presented to aid in understanding the present invention. However, the following examples are merely provided to facilitate understanding of the present invention, and the contents of the present invention are not limited to the following examples.

[0028] The present invention relates to the use of LGI3 as a biomarker for diagnosing the severity of arteriosclerosis.

[0029] More specifically, The present invention provides a biomarker composition for diagnosing the severity of atherosclerosis, comprising LGI3 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3.

[0030] The present invention also provides a biomarker composition for diagnosing the severity of arteriosclerosis, comprising a preparation for measuring the expression or activity level of LGI3 represented by the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3.

[0031] The present invention also provides a kit for diagnosing the severity of arteriosclerosis, comprising the composition.

[0032] The present invention also provides a method for providing information for diagnosing the severity of arteriosclerosis, comprising the steps of: (a) determining the expression or activity level of LGI3 (leucine rich repeat LGI family member 3) represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a sample; and (b) a step of diagnosing the severity of arteriosclerosis as severe if the expression or activity level in step (a) is increased compared to the expression or activity level of LGI3 shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a control group.

[0033] In the present specification, "atherosclerosis" refers to a vascular disease associated with the formation of atheroma, and is also called atherosclerosis or atherosclerosis, and is prevalent in, for example, the coronary artery, carotid artery, inferior abdominal aorta, popliteal artery, etc., but is not limited thereto. The "atherosclerosis" includes, but is not limited to, coronary artery disease and peripheral artery disease. It also includes, but is not limited to, arrhythmia, angina pectoris, myocardial infarction, heart failure, stroke, etc.

[0034] In the present specification, "coronary artery disease (CAD)" refers to a disease in which the blood vessels supplying blood to the heart, particularly the coronary arteries, become clogged or narrowed, resulting in insufficient blood supply to the heart muscle.

[0035] In the present specification, "peripheral artery disease (PAD)" refers to occlusive diseases occurring in the peripheral arteries of the entire body, excluding the coronary arteries and aorta.

[0036] In the present specification, "angina pectoris" refers to chest pressure or chest pain caused by a temporary lack of blood supply to the heart muscle. It is one of the coronary artery diseases that occurs when the coronary artery that supplies blood to the heart is partially narrowed, and is divided into stable angina pectoris, unstable angina pectoris, and variant angina pectoris (Prinzmetal angina pectoris), etc. Stable angina pectoris is mainly caused by arteriosclerosis, etc., and pain mainly occurs when exercising or overeating, when emotionally anxious or excited, or when suddenly exposed to cold air after going out of the house in cold winter. Unstable angina pectoris occurs when the patient is resting or sleeping at night, and the frequency and intensity of existing chest pain gradually increases, and there is a risk of heart failure, so if unstable angina pectoris is suspected, it is necessary to undergo a medical examination and examination and take appropriate measures. Variant angina occurs when the coronary arteries contract like a spasm, causing chest pain. The pain often occurs while sleeping at night or at dawn, but is relieved during daytime activities. It can lead to myocardial infarction and sudden death, and is more common in men, with smoking being an important risk factor.

[0037] In the present specification, "myocardial infarction" refers to a condition in which the blood supply to the heart becomes insufficient, and at one moment the blood vessels become completely blocked by a blood clot, causing blood to stop flowing, resulting in the destruction of a part of the heart muscle and death. Most sudden deaths from heart failure are caused by myocardial infarction.

[0038] In the present specification, "heart failure" refers to a condition in which the heart's pumping action is reduced and the heart is unable to send the necessary amount of blood to other organs in the body, and many heart diseases can cause heart failure by impairing the heart's ability to contract and relax. Heart failure usually progresses slowly over several years, and the heart gradually loses its ability to pump.

[0039] In the present specification, "stroke" refers to brain damage caused by cerebral blood flow disorder, and is classified into hemorrhagic stroke, thrombotic stroke, and embolic stroke. Hemorrhagic stroke refers to brain damage caused by bleeding in the brain parenchyma or between the brain and the skull, which causes blood vessels around the bleeding site to constrict due to spasm, resulting in insufficient blood flow to the surrounding area. Thrombotic stroke occurs when a blood clot formed in a cerebral blood vessel blocks blood flow, usually in an artery narrowed by arteriosclerosis in which fatty components stick to the vascular wall. Embolic stroke occurs when a blood clot or small mass travels along a blood vessel to the brain and blocks the cerebral blood vessel, usually due to a blood clot formed by slowed blood flow in the heart.

[0040] In the present specification, "severity of arteriosclerosis" may mean the degree of increase or decrease in the risk of developing arteriosclerosis, the degree of progression of lesions, and the degree of increase or decrease in the risk of recurrence. The degree of increase or decrease in the risk of developing arteriosclerosis means the degree of risk or possibility of developing arteriosclerosis. The degree of progression of lesions means the degree of minor to serious arteriosclerosis caused by the onset of arteriosclerosis and the progression of lesions. In addition, the degree of increase or decrease in the risk of recurrence means the degree of risk of recurrence or the degree of possibility of recurrence after the determination of complete cure of arteriosclerosis. The severity of arteriosclerosis may be indicated qualitatively and / or quantitatively, and the severity may be classified according to its degree.

[0041] The method for diagnosing the severity of arteriosclerosis according to the present invention may be used in place of a method for diagnosing the severity known in the art, or may be used in parallel with a method for diagnosing the severity known in the art. For example, it is known in the art to measure the SYNTAX (the Synergy between PCI with Taxus and Cardiac Surgery) score to diagnose the severity of coronary artery disease, and the higher the SYNTAX score, the more severe the severity of coronary artery disease. According to a specific embodiment of the present invention, it has been confirmed that the expression level of LGI3 and the SYNTAX score are directly correlated. Also, for example, it is known in the art to measure the Trans-Atlantic Inter-Society Consensus (TASC) to diagnose the severity of peripheral artery disease, and the higher the ratio classified as TASC, the more severe the severity of peripheral artery disease. In the art, it is determined that the TASC classification is from TASC A to TASC D, and the more severe the disease. According to a specific embodiment of the present invention, it has been confirmed that the expression level of LGI3 and the ratio classified as TASC are directly correlated.

[0042] In the present specification, "diagnosis" means to confirm the presence or characteristics of a pathological condition. The diagnosis includes not only the presence or absence of disease, but also prognosis, progress of arteriosclerosis, stage of disease, etc. For the purpose of the present invention, diagnosis means to accurately distinguish and diagnose the severity of arteriosclerosis.

[0043] In the present invention, "LGI3 (leucine rich repeat LGI family member 3)" is used as a biomarker for diagnosing the severity of arteriosclerosis. The LGI3 is not limited to LGI3 of an animal having the protein, and may refer to, for example, human or mouse LGI3. Preferably, the LGI3 amino acid sequence used in the present invention can be represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. The LGI3 that can achieve the object of the present invention includes the whole or a fragment thereof. It also includes a protein or peptide having homology with the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. Here, "peptide" refers to a linear molecule formed by binding amino acid residues to each other by peptide bonds. The peptide derived from LGI3 can be obtained by fragmenting LGI3 protein, and can be produced by chemical synthesis methods known in the art, particularly solid-phase synthesis techniques or liquid-phase synthesis techniques.

[0044] In the present specification, the term "biomarker" refers to a substance that can be used as a diagnostic marker or a diagnostic marker and can classify and diagnose the severity of arteriosclerosis in a sample.

[0045] For the purposes of the present invention, an atherosclerosis diagnostic biomarker of the present invention means that LGI3 shows a specifically higher level of expression or activity in samples from patients with atherosclerosis compared to normal controls; that LGI3 shows a specifically higher level of expression or activity in samples from patients with severe atherosclerosis compared to samples from patients with mild atherosclerosis; or that LGI3 shows a specifically higher level of expression or activity in samples from patients with a high possibility of atherosclerosis recurrence compared to samples from patients with a low possibility of atherosclerosis recurrence.

[0046] The "atherosclerosis severity diagnostic kit" according to the present invention may include not only a preparation for measuring the expression or activity level of LGI3, but also tools, reagents, etc. commonly used in the field for analysis. Examples of the tools or reagents may include, but are not limited to, a suitable carrier, a labeling substance capable of generating a detectable signal, chromophores, a dissolving agent, a detergent, a buffer, a stabilizer, etc. When the labeling substance is an enzyme, a substrate capable of measuring the enzyme activity level and a reaction stopper may be included. The carrier may be a soluble carrier or an insoluble carrier. An example of a soluble carrier is a physiologically acceptable buffer solution known in the field, such as PBS, and an example of an insoluble carrier may be a polymer such as polystyrene, polyethylene, polypropylene, polyester, polyacrylonitrile, fluororesin, cross-linked dextran, polysaccharide, latex plated with metal, or other polymers such as paper, glass, metal, agarose, and combinations thereof.

[0047] Hereinafter, each step of the method for providing information for diagnosing the severity of arteriosclerosis according to the present invention will be described step by step. However, as long as the object of the present invention can be achieved, the following steps may be modified, added, or substituted.

[0048] "(a) confirming the expression or activity level of LGI3 (leucine rich repeat LGI family member 3) represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a sample"

[0049] In the present specification, the term "sample" refers to, but is not limited to, tissue, blood, serum, plasma, urine, saliva, etc., containing LGI3 of a subject (patient) for diagnosing the severity of arteriosclerosis.

[0050] In the present invention, a preparation for measuring the expression or activity level of LGI3 is used to confirm the expression or activity level of LGI3.

[0051] The "preparation for measuring the expression or activity level of LGI3" refers to a substance that can be used to detect LGI3 by specifically binding to the LGI3 protein or the mRNA encoding it and confirming the expression or activity level of LGI3. For example, it can be an oligopeptide, monoclonal antibody, polyclonal antibody, chimeric antibody, ligand, PNA (peptide nucleic acid), aptamer, antisense oligonucleotide, primer pair, or probe that specifically binds to LGI3, but is not limited thereto.

[0052] The term "antibody" is a term well known in the art and means a specific protein molecule that is labeled with an antigenic site. For the purposes of the present invention, an antibody means an antibody that specifically binds to a marker of the present invention, and such an antibody can be produced by cloning each gene into an expression vector by a conventional method to obtain a protein encoded by the marker gene, and then producing the resulting protein by a conventional method. This also includes partial peptides that can be produced from the protein, and the partial peptides of the present invention contain at least 7 amino acids, preferably 9 amino acids, and more preferably 12 or more amino acids. The form of the antibody of the present invention is not particularly limited, and the antibody of the present invention includes polyclonal antibodies, monoclonal antibodies, and any part thereof that has antigen-binding ability, and includes all immunoglobulin antibodies. Furthermore, the antibody of the present invention also includes special antibodies such as humanized antibodies. The antibody used to detect the marker of the present invention includes not only the complete form having two full-length light chains and two full-length heavy chains, but also functional fragments of the antibody molecule. The functional fragment of the antibody molecule means a fragment that has at least antigen-binding function, and includes Fab, F(ab'), F(ab')2, Fv, etc.

[0053] The "primer" refers to a short nucleic acid sequence having a short free 3' hydroxyl group, capable of forming a base pair with a complementary template and serving as a starting point for template strand copying. The primer can initiate DNA synthesis in the presence of a reagent for polymerization reaction (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates at an appropriate buffer solution and temperature. In the present invention, PCR amplification can be performed using sense and antisense primers of the polynucleotide of LGI3, and diagnosis can be made based on the presence or absence of the desired product. The PCR conditions and the lengths of the sense and antisense primers can be modified based on those known in the art.

[0054] The "probe" refers to a nucleic acid fragment such as RNA or DNA, which is composed of several to several hundred bases and can specifically bind to mRNA, and is labeled to confirm the presence or absence of a specific mRNA. The probe may be prepared in the form of an oligonucleotide probe, a single stranded DNA probe, a double stranded DNA probe, an RNA probe, or the like. In the present invention, hybridization is performed using a probe complementary to LGI3, and diagnosis can be made based on the presence or absence of hybridization. Selection of an appropriate probe and hybridization conditions can be modified based on those known in the art.

[0055] The primer or probe of the present invention can be chemically synthesized using phosphoramidite solid support method or other widely known methods. Such nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, capping, substitution of one or more natural nucleotides with analogs, and internucleotide modifications, such as uncharged linkages (e.g., methylphosphonates, phosphotriesters, phosphoramidites, carbamates, etc.) or charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.).

[0056] In the specification of the present invention, "measuring the expression or activity level of LGI3" may refer to the process of confirming the presence or absence and expression or activity level of LGI3 protein or the mRNA encoding it in a sample in order to diagnose arteriosclerosis, and for example, a method of confirming the amount or activity level of the protein or the mRNA encoding it using a molecule that specifically binds to the LGI3 protein or the mRNA encoding it may be applied.

[0057] Methods for confirming the expression or activity level of LGI3 protein include, but are not limited to, Western blot, ELISA (enzyme linked immunosorbent assay), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, and protein chip. In addition, methods for confirming the expression or activity level of mRNA encoding the LGI3 protein include, but are not limited to, reverse transcriptase polymerase reaction (RT-PCR), competitive reverse transcriptase polymerase reaction (competitive RT-PCR), real time RT-PCR, real time quantitative RT-PCR, RNase protection method, Northern blotting, and gene chips.

[0058] "(b) a step of diagnosing the severity of arteriosclerosis as being severe when the expression or activity level in the step (a) is increased compared to the expression or activity level of LGI3 represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 in a control group."

[0059] In the present invention, the expression or activity level of LGI3 is confirmed as described above, and if the expression or activity level of LGI3 is increased compared to a control group, the severity of arteriosclerosis is diagnosed as being severe.

[0060] In the step (b), the term "control group" includes a non-atherosclerotic normal control group, a sample group taken at a different time from the same patient, or a sample group taken from another atherosclerotic patient.

[0061] In the present specification, "the severity of arteriosclerosis is severe" means that the severity of arteriosclerosis is at a high level.

[0062] According to the present invention, when the expression or activity level of LGI3 is increased compared to the control group, the severity of arteriosclerosis is diagnosed as serious. For example, when the expression or activity level of LGI3 is increased in the sample of an arteriosclerosis patient compared to a normal control group, not arteriosclerosis, by control group, the severity of arteriosclerosis is diagnosed as serious. Also, when the expression or activity level of LGI3 is increased in the sample group collected in the embodiment of the present method compared to the sample group collected at another time from the same patient, the severity of arteriosclerosis is diagnosed as serious. Here, the term "another time" may include the time before the start of treatment of an arteriosclerosis patient, during treatment, after complete recovery after treatment, and the time when recurrence may occur after complete recovery. Also, when the expression or activity level of LGI3 is increased in the sample group collected in the embodiment of the present method compared to the sample group collected from another arteriosclerosis patient, the severity of arteriosclerosis is diagnosed as serious.

[0063] The present invention also provides a method for screening a substance for preventing or treating arteriosclerosis, comprising the steps of: (a) confirming the expression or activity level of LGI3 (leucine rich repeat LGI family member 3) shown in the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 after treatment with the candidate substance; and (b) selecting the candidate substance as a substance for preventing or treating arteriosclerosis if the expression or activity level in (a) is downward.

[0064] According to this method, a therapeutic or preventive substance can be appropriately selected depending on the severity of arteriosclerosis.

[0065] Hereinafter, each step of the method for screening a substance for preventing or treating arteriosclerosis according to the present invention will be described step by step. However, since the method includes the step of confirming the expression or activity level of LGI3 as described above, the description of the overlapping content will be omitted to avoid excessive complexity in this specification. And, as long as the object of the present invention can be achieved, the following steps can be modified, added, or replaced.

[0066] "(a) confirming the expression or activity level of LGI3 represented by the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 after treatment with a candidate substance"

[0067] In the present specification, a "candidate substance" may be a nucleotide, DNA, RNA, amino acid, aptamer, protein, chemical compound, natural product, natural extract, etc., and is not limited thereto so long as it is a substance that can be used for the prevention or treatment of arteriosclerosis.

[0068] "(b) if the expression or activity level in the step (a) is decreased, selecting the candidate substance as a substance for preventing or treating arteriosclerosis."

[0069] In the present specification, a downward trend in the expression or activity level of LGI3 can be seen as a decrease in the severity of arteriosclerosis (a downward trend).

[0070] In the present specification, "prevention" refers to any action that inhibits arteriosclerosis or delays the onset of the disease.

[0071] In the present specification, "treatment" refers to any action by which the symptoms of arteriosclerosis are ameliorated or beneficially altered.

[0072] The present invention also provides an use for preventing, ameliorating and / or treating arteriosclerosis, which comprises an LGI3 antagonist having the amino acid sequence of SEQ ID NO: 3 (DEGRQKFVRFQELAV) as an active ingredient.

[0073] More specifically, the present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis, which contains as an active ingredient an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3. The present invention also provides a method for treating arteriosclerosis by administering an effective amount of an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 to an arteriosclerosis patient. The present invention also provides a use of the LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 for treating or preventing arteriosclerosis.

[0074] In particular, the LGI3 antagonist of the present invention does not affect the metabolic phenotype and is effective only in treating arteriosclerosis, allowing treatment without side effects.

[0075] Before preventing and / or treating arteriosclerosis using the LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 according to the present invention, a step of confirming arteriosclerosis disease and / or severity according to the present invention described above may be carried out.

[0076] In the present specification, the term "antagonist" is understood as including all molecules that partially or completely block, inhibit or neutralize one or more of the biological activities of a target. An antagonist can act by decreasing receptor phosphorylation or by incapacitating or killing cells activated by a ligand through the binding of the receptor to the ligand. An antagonist can also completely disrupt the receptor-ligand interaction or substantially reduce the interaction by changing the tertiary structure of the receptor or by down-regulating it.

[0077] In the present invention, the pharmaceutical composition contains an LGI3 antagonist as an active ingredient and may further contain one or more substances commonly used for the prevention or treatment of arteriosclerosis, and may be formulated or used in combination with drugs such as antihistamines, anti-inflammatory analgesics, anticancer drugs, and / or antibiotics.

[0078] In the present invention, the pharmaceutical composition may further comprise suitable carriers, excipients and diluents that are commonly used in the manufacture of pharmaceutical compositions.

[0079] In the present invention, the term "carrier", also called a vehicle, refers to a compound that facilitates the addition of a compound into cells or tissues. For example, diethylsulfoxide (DMSO) is a commonly used carrier that facilitates the introduction of many organic compounds into cells or tissues of living organisms.

[0080] In the present invention, a "diluent" is defined as a compound that is diluted with water to dissolve the compound as well as stabilize the biologically active form of the compound of interest. Salts dissolved in buffer solutions are used as diluents in the art. A commonly used buffer solution is phosphate buffered saline, as it mimics the salt conditions of human solutions. Since buffer salts can control the pH of the solution at low concentrations, it is unusual for a buffer diluent to alter the biological activity of the compound. The compounds used herein can be administered to human patients as such, or as pharmaceutical compositions mixed with other ingredients such as in combination therapy, or with suitable carriers or excipients.

[0081] Additionally, the composition according to the invention may further comprise one or more substances or compounds having therapeutic activity against arteriosclerosis.

[0082] In addition, the pharmaceutical composition according to the present invention can be formulated and used in the form of external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and sterile injection solutions by a conventional method, and the carriers, excipients, and diluents that can be contained in the composition include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to water and liquid paraffin, which are frequently used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, etc. can be used.

[0083] The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of the disease, the drug form, the administration route and period, but can be appropriately selected by those skilled in the art. The administration can be administered once a day or in several divided doses. The dosage does not limit the scope of the present invention in any aspect. In addition, the effective amount for one administration can be formulated in a single preparation in a unit dosage form, formulated in appropriate amounts, or prepared in a multi-dose container.

[0084] The pharmaceutical composition according to the present invention can be administered to mammals such as mice, rats, livestock, and humans by various routes such as parenteral and oral, and all modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine, intradural or intracerebroventricular injection. However, since proteins or peptides are digested when administered orally, oral compositions must be formulated to coat the active agent or protect it from degradation in the stomach. Preferably, it can be administered into the descending thoracic aorta. Alternatively, the composition can be administered by any device that allows the active agent to move to the target cells.

[0085] In the case of oral dosage forms, the dosage may vary depending on the age, sex, and weight of the patient, but may be administered in an amount of 0.1 to 100 mg / kg once or several times a day. The dosage may be increased or decreased depending on the administration route, severity of the disease, sex, weight, age, etc. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

[0086] When the present invention is provided as a mixture in which other ingredients are added in addition to the LGI3 antagonist, the composition may contain the LGI3 antagonist in an amount of 0.001 wt% to 99.9 wt%, preferably 0.1 wt% to 99.0 wt%, and more preferably 30 wt% to 50 wt%, based on the total weight of the composition.

[0087] The present invention provides a food composition for preventing or improving arteriosclerosis, comprising an LGI3 antagonist as an active ingredient. The LGI3 antagonist may be added to food for the purpose of improving arteriosclerosis or vascular disease. When the LGI3 antagonist of the present invention is used as a food additive, the LGI3 antagonist may be added as it is or may be used together with other foods or food ingredients, and may be used appropriately by a conventional method. However, since proteins or peptides are digested when orally administered as food, the oral composition must be formulated so that the active agent is coated or protected from degradation in the stomach. The amount of the active ingredient to be mixed may be determined to suit the purpose of use (prevention, health, or therapeutic treatment). In general, the LGI3 antagonist of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material during the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range.

[0088] In the present invention, the food includes functional food and health functional food, and "functional food" means food in which the functionality of a general food is improved by adding the LGI3 antagonist (Azelaic Acid) of the present invention to the general food. Functionality can be broadly classified into physical properties and physiological functionality, and when the LGI3 antagonist (Azelaic Acid) of the present invention is added to a general food, the physical properties and physiological functionality of the general food are improved, and the present invention defines such foods with improved functions as "functional foods".

[0089] The functional food of the present invention can be used in various ways, such as medicines, foods, and beverages for preventing or improving arteriosclerosis. There is no particular limit to the type of food. Examples of foods to which the substance can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, energy drinks, alcoholic beverages, and vitamin complexes, including all foods in the usual sense.

[0090] The health drink composition according to the present invention may contain various flavorings or natural carbohydrates as additional ingredients, like ordinary drinks. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrates is generally about 0.01 to 20 g, preferably about 5 to 12 g, per 100 mL of the composition of the present invention.

[0091] In addition to the above, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. Such ingredients may be used independently or in combination. The ratio of such additives is not of great importance, but is generally selected in the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

[0092] The present invention can be modified in various ways and can have various embodiments, and a specific embodiment will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to a specific embodiment, and it should be understood that the present invention includes all modifications, equivalents, and alternatives within the spirit and technical scope of the present invention. When describing the present invention, if it is determined that a detailed description of related publicly known technology may make the gist of the present invention unclear, the detailed description will be omitted. EXAMPLES

[0093] [Examples and Experimental Examples] [Example 1] Example 1-1. Experimental animals LGI3 knockout mice (Korea) produced by Macrogen were donated by Dr. Yoon Hyeyoung (Chung-Ang University, Korea). - / - The mice were further bred for >10 generations on a C57B1 / 6 background to confirm that they had no significant abnormalities in spleen and thymus weights or other hematological parameters. - / - ) Mouse strain was purchased from Jackson Laboratory. LGI3 - / - Apoe Mouse - / - Mating with LGI3 mice - / - Apoe - / - We created LGI3 mice that were fed a high-fat, high-cholesterol diet for 16 weeks. - / - Apoe - / - Mice and congenital (congenic) Apoe - / - Mice were used as a control group.

[0094] Example 1-2. Plaque formation test Plaque formation was induced in male littermates (8 weeks old, n=13-15) by feeding a high-fat diet for 16 weeks. Briefly, after mice were euthanized with CO2, the right atrium was removed, and the atrium and aorta were perfused with phosphate-buffered saline (PBS) through the left ventricle. Hearths were inserted into an OCT compound (Sakura, USA) and frozen at -80°C. Aortas were dissected from the proximal ascending aorta to the iliac artery bifurcation, and the adventitial fat was removed. For en face analysis, aortas were opened longitudinally and fixed in 10% neutral buffered formaldehyde in PBS overnight on a flat black silicon plate. Fixed aortas were stained with Oil Red O overnight, washed with PBS, and then digitally photographed at fixed magnification. Total aortic area and lesion area were measured using an Axio Vision microscope. (Carl Zeiss, Germany).

[0095] Examples 1-3. Immunoblotting and ELISA Mouse plasma was measured with a commercially available ELISA kit (LGI3 (CUSABIO CSB-EL012900MO)) according to the manufacturer's instructions.

[0096] Example 1-4. Mouse LGI3 antagonist peptide (LGI3 P34) injection The mouse LGI3 antagonist peptide, LGI3 P34, is a 15-mer synthetic peptide containing amino acids (aa) 1-34 (DEGRQKFVRFQELAV) of mouse Lgi3. The synthetic LGI3 peptide was dissolved in PBS and filter-sterilized. PBS injections were used as the control group for mouse experiments. All peptides were synthesized by Peptron (Seoul, Korea). Apoe mice fed a high-fat diet for 16 weeks were - / - Mice were injected intraperitoneally with PBS or LGI3 antagonist peptide twice a week for the final 5 weeks.

[0097] Examples 1-5. Statistical Analysis Statistical analysis was performed using InStat. Two-tailed Mann-Whitney U-test was used for comparisons between two groups, and one-way ANOVA was used for comparisons of three or more groups. All data are presented as mean ± sem, and a P-value of ≤ 0.05 was considered statistically significant.

[0098] [Example 2] Example 2-1. Study population A total of 68 patients in the database of the Cardiovascular Genetics Center at Yonsei University were included in this study. Patients were enrolled when they underwent coronary angiography for chest pain, peripheral angiography for screening tests, leg claudication (pain that occurs during exercise) or pain during stable disease when they underwent coronary angiography. Subjects aged 30-70 years with ≥50% stenosis in at least one epicardial coronary artery were included.

[0099] Example 2-2. Clinical and Angiographic Data Collection At the time of enrollment, subjects were interviewed regarding their individual medical history and underwent a complete physical examination. Hypertension was defined as blood pressure >140 / 90 mmHg on two or more occasions or antihypertensive therapy. Diabetes was defined as fasting plasma glucose ≥126 mg / dL, postprandial plasma glucose ≥200 mg / dL, or current use of hypoglycemic medication. Hyperlipidemia was defined as low-density low-protein cholesterol ≥160 mg / dL. Coronary artery disease (CAD) and peripheral artery disease (PAD) characteristics were assessed by cardiologists blinded to the study objectives by reviewing coronary and peripheral angiograms. Cardiologists first assessed the clinical expression of the most severe CAD in the patient's medical history, and then assessed the number of coronary arteries with at least 50% stenosis. To assess the severity and extent of CAD, a SYNTAX (PCI with Taxus and Cardiac Surgery) score was determined. The presence of PAD was defined as the presence of at least 50% stenosis from the aorta to the iliac artery level, and the severity was determined by the Trans-Atlantic Inter-Society Consensus (TASC) II definition.

[0100] Example 2-3. ELISA for measuring LGI3 levels in human serum Data on general chemistry and other cardiovascular markers were obtained from the Cardiovascular Genetics Center database. For LGI3 level measurements, venous samples collected after a 12-h fasting period were used. The samples were collected at the time of subject enrollment, centrifuged, and stored at -80°C. For ELISA analysis, human serum was measured using commercially available ELISA kits LGI3 (CUSABIO CSB-EL012900HU), ADIPONECTIN (R&D Systems DRP300) according to the manufacturer's instructions. 96-well plates were coated with 1 mg / well capture antibody using an ELISA kit (Cusabio Biotech Corporation, USA). The coated plates were washed with PBS containing 0.05% Tween 20, incubated with culture medium, and then incubated with biotin-conjugated secondary antibodies. Plates were read at absorbance at 450 nm. Target proteins were analyzed according to the manufacturer's specifications. Appropriate specificity controls were included and all samples were analyzed in duplicate.

[0101] Examples 2-4. Clinical follow-up observations Clinical follow-up was assessed through mandatory record review after discharge, performed at clinical visits. Clinical events were defined by the Academic Research Consortium and Expert Consensus Document, which defined the third universal definition for myocardial infarction. All deaths were considered cardiac-related unless an obvious noncardiac cause was identified. After hospital discharge, myocardial infarction was defined by the presence or absence of clinical symptoms, electrocardiogram changes, or nonnormal imaging findings indicative of myocardial infarction and creatine kinase myocardial band fraction exceeding the upper normal limit or troponin-T / troponin-I exceeding the 99th percentile of the upper limit. Stroke was confirmed by a neurologist based on imaging studies as soon as neurological deficits were demonstrated. Major adverse cardiac and cerebrovascular events (MACCE) were defined as a composite of cardiac death, stroke, and myocardial infarction.

[0102] [Experimental Example 1] Experimental Example 1-1. LGI3 deficiency improved atherosclerotic plaque formation (Figure 1). To investigate the effect of LGI3 deletion on atherosclerotic plaques, we analyzed plaque formation after feeding mice a high-fat diet for 16 weeks. - / - Apoe - / - Mouse Apoe - / - The entire aorta showed reduced plaque formation in mice lacking LGI3 (19.7 ± 2.17, n = 13 vs. 13.34 ± 1.26, n = 15; %, P = 0.0008), and the ascending aorta and arch, abdominal aorta, and especially the descending thoracic aorta each showed greatly reduced plaque formation in mice lacking LGI3.

[0103] Experimental Example 1-2. Deficiency of LGI3 improved aortic lesion formation (Figure 2) LGI3 in the aortic sinus - / - Apoe - / - The mean lesion area in treated mice was significantly lower than that in control mice (103484.9 ± 15558.77, n = 12 vs. 78329.36 ± 14425.65, n = 14; μm 2 , P=0.0017).

[0104] Experimental Example 1-3: The effect of LGI3 on ​​the pathogenesis of atherosclerosis was confirmed (FIG. 3).

[0105] In this study, to investigate the effect of LGI3 on ​​the pathogenesis of atherosclerosis, we used Apoe - / - LGI3 with littermate controls - / - Apoe - / - During the 16-week dietary period starting at 8 weeks of age, weight changes were significantly greater in Lgi3-deficient Apoe mice. - / - It was even smaller in mice. Even more interestingly, Lgi3 - / - Apoe - / - After 16 weeks on a high-fat diet, mice showed a further reduction in inguinal fat mass with a reduced rate of body weight gain compared to controls. These data suggest that Lgi3 deficiency represents a phenotype that reduces disease susceptibility during the pathogenesis of atherosclerosis.

[0106] Experimental Example 1-4. LGI3 was increased in the plasma of an arteriosclerosis mouse model (Figure 4).

[0107] LGI3 is a soluble form proposed as an adipokine and is mainly known as a 60 kDa protein. It has been reported that LGI3 negatively regulates adiponectin and has a positive relationship with tumor necrosis factor-α (TNF-α). TNF-α is a well-known proinflammatory adipokine, and adiponectin is known as a representative anti-inflammatory adipokine. Recently, it has been reported that adipokines released from perivascular adipose tissues affect hyperlipidemia-induced diseases such as atherosclerosis. However, the physiological or pathological relationship between LGI3 and atherosclerosis is unknown. LGI3 expression was increased in white adipose tissues (WAT) of atherosclerotic mice, suggesting that Apoe deficient (Apoe - / - Plasma LGI3 levels were assessed in mice fed a high-fat diet for 13 weeks and were found to be secreted in WAT and to be positively associated with disease severity. - / - Mice were 8 weeks old and - / - Compared with control mice, Apoe+ mice showed higher plasma LGI3 levels (127.31 ± 15.19; n = 4; vs. 33.8 ± 29.42; n = 5, pg / ml, P = 0.049). - / - Conclusion: Plasma LGI3 levels were significantly increased in mice. These data suggest that LGI3 plays an important role in atherosclerosis and that LGI3 is a biomarker for atherosclerosis in mice.

[0108] Experimental Example 1-5. LGI3 antagonist inhibits Apoe - / - It reduced atherosclerotic plaque formation in mice (Figure 5).

[0109] Our data established that LGI3 is a novel biomarker in chronic inflammatory diseases, especially atherosclerosis. Based on the premise that LGI3 is secreted from WAT when atherosclerotic conditions are induced and promotes proinflammatory signaling through a mechanism that is not known to be mediated by its receptors, ADAM22 or ADAM23, we administered PBS or an LGI3 antagonist peptide to Apoe+ mice fed a high-fat diet for 16 weeks to test the effect of LGI3 inhibition and its potential for developing into a therapeutic target. - / - Mice were intraperitoneally injected with LGI3 antagonist peptide. Interestingly, the LGI3 antagonist peptide-injected group showed reduced plaque formation compared to the control group (11.42 ± 0.97; n = 9 vs. 14.76 ± 1.26; n = 9; %, P = 0.04). Notably, plaque formation was significantly reduced after treatment with LGI3 antagonist in the descending thoracic aorta (4.44 ± 0.58; n = 9 vs. 7.52 ± 1.26; n = 9; %, P = 0.01), suggesting that LGI3 can act on the receptor ADAM23, which is expressed by perivascular adipose tissue located in close proximity to the aorta. These data also demonstrated that LGI3 inhibition could be developed as a therapeutic strategy to treat atherosclerosis.

[0110] Experimental Example 1-6. LGI3 antagonists exhibited plaque reducing effects without altering the metabolic phenotype (FIG. 6).

[0111] Body weight and weight of other major metabolic organs, especially epididymal fat weight, showed no difference between PBS-injected and LGI3 antagonist-injected groups (39.22 ± 3.51; n = 9 vs. 38.08 ± 2.19; n = 9; g, P = 0.536 1.689 ± 0.43; n = 9 vs. 1.58 ± 0.51; n = 9; %, P = 0.84). These data suggested that the effect of LGI3 antagonist peptide was predominant on the formation of atherosclerotic plaques, rather than reducing fat or alleviating obesity. These data also indicated that LGI3 antagonist peptide could improve atherosclerosis without changing metabolic parameters and could be used as a therapeutic agent for atherosclerosis.

[0112] [Experimental Example 2] The mean LGI3 level was 13.04 ± 8.03 ng / mL, and patients were classified according to LGI3 level quartiles as follows: Q1 (n = 17) 4.34 ± 1.58 ng / mL, Q2 (n = 17) 9.79 ± 1.53 ng / mL, Q3 (n = 17) 14.19 ± 1.82 ng / mL, and Q4 (n = 17) 23.85 ± 6.96 ng / mL. Clinical and laboratory findings according to LGI3 level quartiles are shown in Table 1. Although not statistically significant, BMI was lower in the LGI3 level quartiles (P = 0.083). Creatine and hsCRP were also higher in the LGI3 level quartiles.

[0113] [Table 1]

[0114] The SYNTAX score gradually increased with the LGI3 level quartile, and was highest when the SYNTAX score was in the LGI3 level quartile (25.9 ± 14.3) (P < 0.001) (left side of Figure 7). In addition, when the LGI3 level was treated as a continuous variable, a positive relationship was confirmed between the LGI3 level and the SYNTAX score (R2 = 0.419, P < 0.001) (right side of Figure 7).

[0115] It was also shown that high LGI3 levels were associated with the degree and severity of CAD. Besides the severity of CAD, the frequency of PAD also gradually increased with LGI3 level quartiles from 5.9%, 5.9%, 11.8%, and 29.4% (Fig. 8). The angiographic extent of PAD defined by the TASCII classification was significantly more severe with LGI3 level quartiles (P=0.036) (Fig. 8).

[0116] During a median follow-up period of 5.7 years (interquartile range 3.9-10.1 years), there were no myocardial infarctions or strokes in patients with LGI3 levels in the first quartile, acute myocardial infarctions in patients with LGI3 levels in the second quartile, strokes in patients with LGI3 levels in the third quartile, and acute myocardial infarctions in patients with LGI3 levels in the fourth quartile. Thus, the 8-year MACCE survival rates were 100%, 93%, 87%, and 68%, respectively (Figure 9), suggesting that the higher the LGI3 level, the higher the frequency of cardiac and cerebrovascular diseases.

[0117] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified in other specific forms without changing the technical concept or essential features of the present invention. Therefore, the above-described embodiments should be understood to be illustrative in all respects and not restrictive. [Industrial Applicability]

[0118] According to the present invention, it is possible to develop a therapeutic substance for improving the severity of arteriosclerosis, and the present invention is expected to provide a stepping stone for utilizing such a therapeutic substance in various fields such as pharmaceutical, topical, food additive compositions, etc. Furthermore, according to the present invention, it is expected that it is possible to treat arteriosclerosis without affecting metabolic phenotype, and therefore it is possible to treat arteriosclerosis without side effects. [Sequence List Free Text]

[0119] SEQ ID NO:1 (leucine rich repeat LGI family member 3) Length:548 Type: Protein Organism name: Homo sapiens array: 1 maglrarggp gpgllalsal gfclmlqvsa krppktppcp pscsctrdta fcvdskavpr 61 nlpsevislt lvnaafseiq dgafshlpll qflllnsnkf tligdnaftg lshlqylfie 121 nndiwalskf tfrglkslth lslannnlqt lprdifrpld ilndldlrgn slncdckvkw 181 lvewlahtnt tvapiycasp prfqehkvqd lplrefdcit tdfvlyqtla fpavsaepfl 241 yssdlylala qpgvsactil kwdyverqlr dydripapsa vhckpmvvds qlyvvvaqlf 301 ggsyiyhwdp nttrftrlqd idpqrvrkpn dleafridgd wyfavadssk agatslyrwh 361 qngfyshqal hpwhrdtdle fvdgegkprl ivssssqapv iyqwsrtqkq fvaqgevtqv 421 pdaqavkhfr agrdsylcls ryigdskilr wegtrfsevq alpsrgslal qpflvggrry 481 lalgsdfsft qiyqwdegrq kfvrfqelav qaprafcymp agdaqlllap sfkgqtlvyr 541 hivvdlsa SEQ ID NO:2 (leucine rich repeat LGI family member 3) Length:548 Type: Protein Organism name:Mus musculus array: 1 maglrarrgp grrllvlstl gfclmlqvsa krppktppcp pscsctrdta fcvdsksvpk 61 nlpsevislt lvnaafseiq dgafshlpll qflllnsnkf tligdnafig lshlqylfie 121 nndiwalskf tfrglkslth lslannnlqt lprdifrpld ilsdldlrgn alncdckvkw 181 lvewlahtnt tvapiycasp prfqehkvqd lplrefdcit tdfvlyqtls fpavsaepfl 241 yssdlylala qpgasactil kwdyverqlr dydripapsa vhckpmvvdg qlyvvvaqlf 301 ggsyiyhwdp nttrftklqd idpqrvrkpn dleafridgd wffavadssk agatslyrwh 361 qngfyshqal hawhrdtdle fvdgegkprl ivssssqapv iyqwsrsqkq fvaqgevtqv 421 pdaqavkhfr agrdsylcls ryigdskilr wegtrfsevq alpsrgslal qpflvgghry 481 lalgsdfsft qiyqwdegrq kfvrfqelav qaprafcymp agdaqlllap sfkgqtlvyr 541 hvvvdlsa SEQ ID NO:3 (leucine rich repeat LGI family member 3) Length:15 Type: Protein Organism name: Homo sapiens Sequence:DEGRQKFVRFQELAV

Claims

1. A pharmaceutical composition for preventing or treating arteriosclerosis, comprising an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO:3 as an active ingredient.

2. 2. The pharmaceutical composition according to claim 1, wherein the arteriosclerosis is coronary artery disease (CAD) or peripheral artery disease (PAD).

3. A food composition for preventing or ameliorating arteriosclerosis, comprising an LGI3 antagonist represented by the amino acid sequence of SEQ ID NO: 3 as an active ingredient.

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