KR102057441B1 - Pharmaceutical composition for preventing or treating immunocyte migration-related diseases comprising benzo[d]thiazole derivatives - Google Patents

Abstract

A benzo [d] thiazole derivative having the structure of Formula 1 relates to the prevention or treatment of diseases related to immune cell migration. The compound of Formula 1 can regulate the migration of immune cells, and thus has a very significant effect in the prevention, improvement and treatment of diseases related to immune cell migration.

Description

Pharmaceutical composition for preventing or treating immunocyte migration-related diseases comprising benzo [d] thiazole derivatives}

The present invention relates to a novel use of benzo [d] thiazole derivatives having the unique structure disclosed herein, and more particularly, to immune cell migration of benzo [d] thiazole derivatives having the structure of Formula (1). A prophylactic or therapeutic use of a related disease.

In many tissues of the body, each cell migrates in different ways depending on their genetic characteristics and environment. Uncontrolled cell migration involves various disease states such as inflammatory disease, cancer metastasis, etc., but the migration signaling and mechanism characteristics of each cell are not fully characterized. In particular, each cell is reported to have a different way of relating to the same factors, which further increases the difficulty in identifying signaling processes and mechanisms. For example, AQP1 (water channel aquaporin-1) promotes cell migration in epithelial cells and the like. Hara-Chikuma M et al., Aquaporin-1 facilitates epithelial cell migration in kidney proximal tubule, J Am Soc Nephrol. 2006 Jan; 17 (1): 39-45 Jiang Y, Aquaporin-1 activity of plasma membrane affects HT20 colon cancer cell migration, IUBMB Life.2009 Oct; 61 (10): 1001-9). (Tyteca D et al., Regulation of Macrophage Motility by the Water Channel Aquaporin-1: Crucial Role of M0 / M2 Phenotype Switch, PLoS One. 2015 Feb 26; 10 (2): e0117398). As these cells have different ways and characteristics in their migration, drugs designed to prevent the migration of specific cells have been shown to be quite limited and insufficient in efficacy. Therefore, there is a need for a new strategy to control the migratory switch (cell) of the cell and to treat migration-related diseases.

On the other hand, although immune cells are also the first defense network in the body, excessive activation of immune cells has recently been reported to be one of the major pathogenesis. In general, an increase in the mobility of immune cells is observed upon activation of inflammatory immune cells. Specifically, it has been reported that such immune cell migration and infiltration are closely related to the pathology of the disease in the following diseases.

Cardiovascular diseases, for example, are diseases of the heart and major arteries, including atherosclerosis and coronary artery disease (Ross R et al., New Engl J Med, 1999: 340 (2): 115-26 , Poli G et al., Redox Biol 2013; 1 (1): 125-30, Libby P et al., Circulation 2002; 5; 105 (9): 1135-43). Atherosclerosis is an inflammatory disease caused by cholesterol and is caused by atherosclerosis consisting of cholesterol deposited on the inner artery membrane and immune cells that migrated from the blood into the artery. In other words, at the site where cholesterol oxides are inflamed, immune cells such as monocytes migrate to form atherosclerosis. When atherosclerosis is formed, the inner surface of blood vessels becomes thick and the wall becomes thick, and the diameter of the inside of the blood flow is narrowed, which causes a problem in blood circulation. When the fibrous membrane around the atheromatous mass bursts, blood clots develop in the blood vessels, and bleeding into the atheromatous mass causes the vessel's internal diameter to narrow rapidly or become blocked. It occurs mainly in blood vessels that supply blood to the heart, blood vessels that supply blood to the brain, blood vessels that supply blood to the kidneys, and peripheral blood vessels, causing ischemic heart disease, ischemic cerebrovascular disease (stroke), kidney failure, and limb ischemic artery disease. . Conventionally, CCL2 (CCChemokine ligand 2, MCP-1), which causes inflammatory reactions by inducing monocyte migration, is known to play an important role in the development and development of cardiovascular diseases. New methods of treating such cardiovascular diseases by inhibiting them have been proposed (Gu L et al., Mol Cell, 1998; 2 (2): 275-81, Aiello RJ et al., Arterioscler Thromb Vasc Biol 1999; 19 (6): 1518-25, Gosling J1 et al., Clin Invest 1999; 103 (6): 773-8, Harrington JR et al., Stem Cells 2000; 18 (1): 65-6, Ikeda U et al., Clin Cardiol 2002; 25 (4): 143-7). In addition, even in high blood pressure, various immune cells that secrete inflammatory cytokines are excessively moved into blood vessels, resulting in thickening of blood vessel walls, and pathologies in which the elasticity of blood vessels is lost.

Pulmonary Arterial Hypertension (PAH) is also classified as Group 1 of the World Health Organization (WHO) Clinical Classification System (ESC Guidelines, European Heart Journal 2015), and is known as dyspnea, mean pulmonary artery pressure (mPAP). ) Is a rare disease with common clinical features of elevation (mPAP> 25 mmHg) and right ventricular dysfunction. Although pulmonary hypertension is involved in several preexisting factors such as heredity, infection, and related diseases, the immune response due to endothelial cell injury is known to be a key pathological factor (Huertas et al., Circulation, 129: 1332). -1340, 2014). In this phenomenon, a series of processes due to infiltration and dysfunction of immune cells are known to be deeply associated with pathology. Especially, PAH interaction is important for PAH. In addition, in recent years, Alport syndrome has been reported to invade monocytes and macrophages to promote the disease progression.

On the other hand, in fibrosis-related diseases, a persistent (chronic) inflammatory response activates a wound-healing program, which leads to fibrosis. After tissue damage, inflammatory immune cells such as monocytes / macrophages, neutrophils, eosinophils, and mast cells are rapidly penetrated into the site of injury, activating and secreting several cytokines, which are the surrounding fibroblasts, epithelial cells, and smooth muscle. The cells are reactivated to activate them into miyoblast cells, which produce and secrete large amounts of extracellular matrix proteins, ultimately causing large accumulations of extracellular matrix proteins in tissues, resulting in wound healing. It also leads to fibrosis or enlargement of tissues (Gurtner GC et al., Trends Cell Biol. 15: 599-607, 2005). This pathological mechanism is one of the fundamental causes of scar formation in the skin tissue caused by wounds, burns, bedsores, etc., and sclerotic fibrosis of tissues such as liver, kidney, blood vessels and lungs. Fibrosis also occurs in chronic autoimmune diseases such as scleroderma, rheumatoid arthritis, Crohn's disease, ulcerative colitis, myelofibrosis, and systemic lupus erythematosus. Is a major pathological feature. In addition, activation of inflammatory immune cells in atopic disease, asthma, COPD, psoriasis, keloids, proliferative retinopathy is known to contribute to the pathology.

 In particular, the fibroblasts activated as miyoblast type cells in the wound-healing program are called myofibroblasts (myofibroblasts). Since myofibroblast is at the center of all fibrosis-related disease pathologies, eliminating the molecular biological or immunological mechanisms that induce myofibroblast activity is a key component of disease treatment. It is well known that many innate or adaptive immunity immunity is important for the activity and differentiation of fibroblasts. Therefore, eliminating inflammatory reactions at the wound site stops tissue remodeling and prevents normal tissue morphology. It is a key element to maintain. In practice, however, the elimination of inflammatory reactions is not easy, so understanding the mechanism of innate immunity and finding key mediators is important in slowing fibrosis.

Monocytes and macrophages contribute to wound healing, but they release reactive oxygen and nitrogen, which in turn cause harmful effects on surrounding cells. Therefore, without the rapid removal of monocytes, macrophages will cause more tissue damage and result in fibrosis. Therefore, limiting monocytes and macrophages that respond first in the early stages of the disease is considered a therapeutic strategy for various chronic inflammatory and fibrotic related diseases.

When the wound-healing mechanism triggers a fibrosis reaction, platelet-derived growth factor (PDGF), which is involved in hemagglutination, recruits other inflammatory immune cells into the wound and TGF-β1 promotes extracellular matrix synthesis from local fibroblasts. It is known to let. However, it has been reported that the factors involved in the hemagglutination reaction induce fibrosis even when they are deficient.

In such diseases in which excessive immune cell activation is a problem, target factors have been suggested to prevent the migration (and invasion) of immune cells, and attempts to devise therapeutic methods for the disease have been proposed. Although the limitations are being reported, it is still important to find out what are the key mediators and strategies for controlling the movement of immune cells for effective disease treatment. There is a situation.

Therefore, while the present inventors have been searching for a new therapeutic strategy for immune cell migration (infiltration) related diseases, the increase of KRS level in the cell membrane region of immune cells (monocytes / macrophages) is associated with immune cell migration and invasion related diseases. It has been shown to be an important pathological phenomenon, which is particularly associated with laminin (particularly, laminin subtype α4β2γ1), and that specific compounds possessing the structure of formula 1 inhibit the migration of KRS to the cell membrane, resulting in increased KRS levels in the cell membrane of immune cells. The present invention was completed by confirming that there is an effect of treating immune diseases by reducing immune cell migration and infiltration.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of immune cell migration-related diseases comprising a compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

<Formula 1>

Where

R1, R2, and R3, independently of each other, are hydrogen; Halogen group; Nitro group; Amino group; C1-C6 alkyl group optionally substituted with halogen; Or a hydroxycarbonyl group, provided that R 1, R 2, and R 3 cannot be hydrogen at the same time,

R4, R5, and R6, independently of each other, are hydrogen; Halogen group; C1-C6 alkyl group; C 1 -C 6 alkoxy group optionally substituted with C 3 -C 6 cycloalkyl; C1-C6 alkylsulfanyl group; Or mono- or di-C1-C6 alkylamino group,

R7 and R8, independently of each other, are hydrogen; Hydroxyl group; Halogen group; Or a hydroxycarbonyl group, provided that R7 and R8 cannot be hydrogen at the same time,

R9 is hydrogen or a C1-C6 alkyl group.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating immune cell migration-related diseases comprising a compound of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient:

<Formula 1>

Where

R1, R2, and R3, independently of each other, are hydrogen; Halogen group; Nitro group; Amino group; C1-C6 alkyl group optionally substituted with halogen; Or a hydroxycarbonyl group, provided that R 1, R 2, and R 3 cannot be hydrogen at the same time,

R4, R5, and R6, independently of each other, are hydrogen; Halogen group; C1-C6 alkyl group; C 1 -C 6 alkoxy group optionally substituted with C 3 -C 6 cycloalkyl; C1-C6 alkylsulfanyl group; Or mono- or di-C1-C6 alkylamino group,

R7 and R8, independently of each other, are hydrogen; Hydroxyl group; Halogen group; Or a hydroxycarbonyl group, provided that R7 and R8 cannot be hydrogen at the same time,

R9 is hydrogen or a C1-C6 alkyl group.

Justice

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The following references provide one of the skills having a general definition of several terms used in the specification of the present invention. Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOTY (2d ed. 1994); THE CAMBRIDGE DICTIONARY OF SCIENCE AND TECHNOLOGY (Walker ed., 1988); And Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY. In addition, the following definitions are provided to assist the reader in practicing the present invention.

In the present invention, the term 'alkyl' refers to an aliphatic hydrocarbon radical, and includes both linear or branched hydrocarbon radicals. For example, C1-C6 alkyl is an aliphatic hydrocarbon having 1 to 6 carbon atoms, methyl, ethyl, propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl , Neopentyl, isopentyl, and the like.

In addition, the term "alkoxy" means a radical in which the hydrogen atom of the hydroxy group is substituted with alkyl unless otherwise defined, for example C1-C6 alkoxy is methoxy, ethoxy, propoxy, n-butoxy, n -Pentyloxy, isopropoxy, sec-butoxy, tert-butoxy, neopentyloxy, isopentyloxy and the like.

By "KRS protein" is meant polypeptides known as lysyl tRNA synthetases. KRS is an enzyme that mediates the aminoacylation reaction of amino acids lysine and tRNA. In the present invention, the specific sequence is not particularly limited as long as KRS is known in the art as a lysyl thioene synthase. For example, the KRS of the present invention is derived from human ( homo sapiens ) and is NCBI (Genbank) Accession No. NP_005539.1 and the like.

Hereinafter, the present invention will be described in detail.

With respect to the pathological migration of cells, the present inventors have found that it is important for KRS to be specifically elevated in the cell membrane of immune cells (especially monocytes / macrophages) relative to the cytoplasm of immune cells. For the first time, we identified the specific regulatory function of KRS in immune cell (monocyte / macrophage) migration, especially with LN421 (laminin subtype α4β2γ1). Therefore, the compound of formula 1 specifically inhibits the migration of KRS from the cytoplasm to the cell membrane, thereby significantly reducing the KRS level of the cell membrane. It has been shown to have a significant therapeutic effect in related diseases.

Therefore, the present invention provides a pharmaceutical composition for preventing or treating immune cell migration-related diseases comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the compounds according to the invention or pharmaceutically acceptable salts thereof, preferably R 1, R 2, and R 3, independently of one another, are hydrogen; Or a halogen group, provided that R1, R2, and R3 cannot be hydrogen at the same time,

R4, R5, and R6, independently of each other, are hydrogen; Halogen group; Or a C1-C6 alkoxy group, provided that R4, R5, and R6 cannot be hydrogen at the same time,

R7 and R8, independently of each other, are hydrogen; Or a hydroxycarbonyl group, provided that R7 and R8 cannot be hydrogen at the same time,

R9 is hydrogen or a C1-C6 alkyl group.

In the compound of Formula 1 of the present invention, preferred compounds or salts thereof are as follows:

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-(((2-chlorophenylethyl) (7-fluorobenzo [d] thiazol-2-yl) amino) methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3-fluorophenyl) ethyl] amino} methyl) benzoic acid;

4-{[[2- (4-chlorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-{[[2- (3-chlorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-(((7-fluorobenzo [d] thiazol-2-yl) (4-methylphenylethyl) amino) methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-fluorophenyl) ethyl] amino} methyl) benzoic acid;

4-{[[2- (4-ethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-propoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-isopropoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) {2- [4- (methylsulfanyl) phenyl] ethyl} amino] methyl} benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) {2- [3- (methylsulfanyl) phenyl] ethyl} amino] methyl} benzoic acid;

4-({[2- (2,5-dimethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (3,4-dimethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (2-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3-methylphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-isobutoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-{[[2- (4-cyclopropylmethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) {2- [4- (methylamino) phenyl] ethyl} amino] methyl} benzoic acid;

4-{[{2- [4- (dimethylamino) phenyl] ethyl} (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) (2-phenylethyl) amino] methyl} benzoic acid;

4-{[[2- (4-cyclohexylmethoxyphenyl) ethyl]-(7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-{[[2- (4-cyclobutylmethoxyphenyl) ethyl]-(7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-({[2- (4-ethoxy-3-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (2-fluoro-4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (2,4-dimethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3-fluoro-4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (3-chloro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-{[[2- (4-sec-butoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-{[[2- (4-ethylaminophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-({[2- (4-ethylphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) {2- [4- (propan-2-yl) phenyl] ethyl} amino] methyl} benzoic acid;

4-({[2- (2,3-difluorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (2,5-difluorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3,4,5-trifluorophenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (3-bromo-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (2,4-dichlorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (2,4-difluorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (2-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (2-fluorophenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (3-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (2,3-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-{[(7-fluorobenzo [d] thiazol-2-yl) {2- [4-methoxy-2-methyl-5- (propan-2-yl) phenyl] ethyl} amino] methyl} Benzoic acid;

4-({[2- (2,5-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (2-chloro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,3-dimethylphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,5-dimethylphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-3-methylphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2-methylphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (2,6-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} propyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (3-fluoro-4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (3-bromo-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (2,4-difluorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (2,3-difluorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (3,4,5-trifluorophenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (2,4-dichlorophenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (2,3-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (2,5-Difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- {1-[(7-fluorobenzo [d] thiazol-2-yl) {2- [4-methoxy-2-methyl-5- (propan-2-yl) phenyl] ethyl} amino] Ethyl} benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,3-dimethylphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,5-dimethylphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-3-methylphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2-methylphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (2,6-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (2,3-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} propyl) benzoic acid;

4- (1-{[2- (2,5-Difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} propyl) benzoic acid;

4- (1-{[2- (2-chloro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} propyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,3-dimethylphenyl) ethyl] amino} propyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2,5-dimethylphenyl) ethyl] amino} propyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-3-methylphenyl) ethyl] amino} propyl) benzoic acid;

4- (1-{(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxy-2-methylphenyl) ethyl] amino} propyl) benzoic acid;

4- (1-{[2- (2,6-difluoro-4-methoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino} propyl) benzoic acid;

4- {1-[(7-fluorobenzo [d] thiazol-2-yl) {2- [4-methoxy-2-methyl-5- (propan-2-yl) phenyl] ethyl} amino] Propyl} benzoic acid;

4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) -2-hydroxybenzoic acid;

3-chloro-4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] (6-nitro-benzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] (7-nitro-benzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({(6-amino-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-amino-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(6-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(5-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(5,6-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-{[[2- (4-methoxyphenyl) ethyl] (5,6,7-trifluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-({(6-chloro-7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-{[[2- (4-methoxyphenyl) ethyl] (7-trifluoromethyl-benzo [d] thiazol-2-yl) amino] methyl} benzoic acid;

4-({(6,7-difluorobenzo [d] thiazol-2-yl)-[2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(5-bromo-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(6-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(5,7-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({(7-fluoro-6-methyl-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] ami

No} methyl) benzoic acid;

4-({(4,6-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] (7-methyl-benzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] (6-methyl-benzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] (5-methyl-benzo [d] thiazol-2-yl) amino} methyl) benzoic acid;

4- (1-{(5,7-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(5,6-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] (5,6,7-trifluorobenzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{(6-chloro-7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(5-bromo-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(6,7-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-fluoro-6-methyl-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] [7- (trifluoromethyl) -benzo [d] thiazol-2-yl] amino} ethyl) benzoic acid;

4- (1-{(5-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(6-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(7-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] (7-methyl-benzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] (6-methyl-benzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] (5-methyl-benzo [d] thiazol-2-yl) amino} ethyl) benzoic acid;

4- (1-{(5-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{(4,6-difluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4-({(6-bromo-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4- (1-{(6-bromo-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

2-{(4-carboxybenzyl) [2- (4-methoxyphenyl) ethyl] amino} benzo [d] thiazole-6-carboxylic acid;

2-{(4-carboxybenzyl) [2- (4-methoxyphenyl) ethyl] amino} benzo [d] thiazole-7-carboxylic acid;

4-({[2- (4-methoxyphenyl) ethyl] [6- (trifluoromethyl) benzo [d] thiazol-2-yl] amino} methyl) benzoic acid;

4-({[2- (4-methoxyphenyl) ethyl] [5- (trifluoromethyl) benzo [d] thiazol-2-yl] amino} methyl) benzoic acid;

4-({[7-fluoro-6- (trifluoromethyl) benzo [d] thiazol-2-yl] [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;

4- (1-{(5,7-difluorobenzo [d] thiazol-2-yl) [2- (3-fluoro-4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] [6- (trifluoromethyl) benzo [d] thiazol-2-yl] amino} ethyl) benzoic acid;

4- (1-{[2- (4-methoxyphenyl) ethyl] [5- (trifluoromethyl) benzo [d] thiazol-2-yl] amino} ethyl) benzoic acid; And

4- (1-{[7-fluoro-6- (trifluoromethyl) benzo [d] thiazol-2-yl] [2- (4-methoxyphenyl) ethyl] amino} ethyl) benzoic acid.

A more preferred example of the compound of Formula 1 in the present invention may be a compound having a structure represented by the formula (1-1).

<Formula 1-1>

In Chemical Formula 1-1,

R 1 is hydrogen; Or a halogen group,

R 4 is hydrogen; Halogen group; Or a C1-C6 alkoxy group

R7 is hydrogen; Or a hydroxycarbonyl group.

In one embodiment of the present invention, among the compounds represented by the formula (1), a compound that shares the structure of the formula (1-1), in particular 4- ({(7-fluorobenzo [d ] Thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid as an example, applied to in vivo disease models for various kinds of immune cell migration (and invasion) related diseases Has confirmed the effectiveness of the disease prevention and treatment. The compound of formula 2 is also referred to herein as 'BC-KI-00053'.

<Formula 2>

The compound of Formula 1 or a salt thereof may have a substituent including an asymmetric atom, in which case the compound of Formula 1 or a salt thereof may exist as an optical isomer such as (R), (S), or racemic (RS). Can be. Therefore, unless otherwise indicated, the compound of Formula 1 or a salt thereof includes all optical isomers such as (R), (S), or racemic (RS).

The compound of formula 1 of the present invention may be in the form of a pharmaceutically acceptable salt. The salts are salts derived from conventional acid addition salts, for example salts derived from inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid or phosphoric acid and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, Salts derived from organic acids such as benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid or aspartic acid. The salts may also be in the form of conventional metal salts, for example alkali metal salts such as lithium, sodium, or potassium; Alkaline earth metal salts such as calcium or magnesium salts; Or chromium salts. Also included are salts formed with suitable organic ligands, such as quaternary ammonium salts, and dicyclohexylamine or N-methyl-D-glucamine salts and amino acid salts formed with arginine, lysine and the like.

The term “immune cell” in the present invention preferably means monocytes or macrophages.

The term "immune cell migration related disease" in the present invention is not particularly limited if the specific immune system is known in the art that excessive immune cell migration (or / and infiltration) is a major pathogenesis, for example, cardiovascular disease, It may be selected from the group consisting of fibrotic disease, inflammatory disease and Alport syndrome.

The specific cardiovascular disease is not particularly limited, but for example, hypertension (including inflammatory complications caused by hypertension), pulmonary arterial hypertension, atherosclerosis, angina pectoris, myocardial infarction, ischemic cerebrovascular disease, arteriosclerosis, and mesenteric sclerosis. It may be selected from the group.

The fibrotic disease is not particularly limited in its specific kind, for example Scleroderma, rheumatoid arthritis, Crohn's disease, ulcerative colitis, myelofibrosis, pulmonary fibrosis, hepatic fibrosis, liver cirrhosis cirrhosis, kidney fibrosis, glomerulosclerosis, myofibrosis, cardiac fibrosis, interstitial fibrosis, pancreatic fibrosis, spleen fibrosis, mesenteric fibrosis, vascular fibrosis, skin fibrosis, eye fiber Fibrosis, macular degeneration, joint fibrosis, thyroid fibrosis, endocardial fibrosis, peritoneal fibrosis, peritoneal fibrosis, progressive mass fibrosis, neoplastic fibrosis, systemic lupus erythematosu, Hereditary fibrosis, infectious fibrosis, irritant fibrosis, fibrosis due to chronic autoimmunity, fibrosis due to antigen incompatibility during organ transplantation, fibrotic complications of surgery, hyperlipidemia Fibrosis, may be selected from fibrosis, diabetes, fibrosis, the group consisting of occlusion due to fibrosis and fibrosis during stenting due to hypertension caused by obesity.

In the present invention, the inflammatory disease is not particularly limited in kind, but preferably, autoimmune disease, inflammatory bowel disease, dermatitis (for example, atopic dermatitis, eczema, psoriasis, etc.), diabetic eye disease (diabetic Retinopathy, peritonitis, osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, rhinitis, sinusitis, otitis media, pneumonia, gastritis, enteritis, cystic fibrosis, stroke (stroke, stroke, etc.), bronchitis, bronchiolitis , Hepatitis (cirrhosis, non-alcoholic steatohepatitis, etc.), nephritis (diabetic renal failure, etc.), arthritis (psoriatic arthritis, osteoarthritis, etc.), neuritis (diabetic neuropathy, multiple sclerosis, etc.), gout, Spondylitis, lighter syndrome, polyarteritis nodular, vasculitis, Lou Gehrig's disease, Wegener's granulomatosis, hypercytokinemia, rheumatoid polymyalgia, arterial cell arteritis, calcium crystal needle Arthrosis, pseudogout, non-articular rheumatism, bursitis, hay fever, epicondylitis (tennis elbow), neuropathic joint disease (Charcot's joint), hemarthrosis, henoch-Scholine purpura, thickening osteoarthritis, multiple Acute reticulocytosis, surcoilosis, hemochromatosis, sickle cell disease and other hemoglobinopathies, hyperlipoproteinemia, hypomagglobulinemia, hyperparathyroidism, acromegaly, familial Mediterranean fever, Beh't disease, systemic erythematosis Lupus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, acute respiratory distress syndrome, multiple organ failure, chronic obstructive pulmonary disease, acute lung injury and bronchial lung dysplasia -pulmonary dysplasia), and also includes chronic inflammatory diseases.

In the present invention the autoimmune disease is Rheumatoid arthritis, systemic scleroderma, systemic lupus erythematosus, psoriasis, asthma, ulcerative colitis, Beche's disease, Crohn's disease, multiple sclerosis, dermatitis, collagen disease, vasculitis, arthritis, granulomatosis, organ specific autoimmune lesions, ulcerative colitis And it may be selected from the group consisting of GvHD (graft-versus-host disease).

The chronic inflammatory disease refers to a condition in which they are chronicized with reference to the aforementioned types of inflammatory diseases, and preferred examples thereof include asthma, atopic dermatitis, eczema, psoriasis, osteoarthritis, gout, Psoriatic arthritis, cirrhosis, alcoholic fatty liver disease, chronic obstructive pulmonary disease, rhinitis, diabetic retinopathy, diabetic renal failure, diabetic neuropathy and multiple sclerosis, but are not limited thereto.

The pharmaceutical composition according to the present invention may contain the compound of Formula 1 or a pharmaceutically acceptable salt thereof alone or in a suitable form with a pharmaceutically acceptable carrier, and further contain an excipient or diluent. can do. As used herein, 'pharmaceutically acceptable' refers to a non-toxic composition that, when administered to human beings, is physiologically acceptable and typically does not cause allergic or similar reactions such as gastrointestinal disorders, dizziness, and the like.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, it may include various drug delivery materials used for oral administration. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent and the like in addition to the above components. Other pharmaceutically acceptable carriers and formulations may be referred to as described in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995.

The composition of the present invention may be administered to any mammal, including humans. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intraperitoneal, intracranial, subcutaneous, intramuscular, intraocular, intraarterial, cerebrospinal, intramedullary, intradural, intracardiac, transdermal, subcutaneous, Injections or infusions by intranasal, intestinal, topical, sublingual, rectal or intralesional routes, or injections or infusions by sustained release systems described below. In one example, the compound of Formula 1 may be administered systemically or locally.

The pharmaceutical composition of the present invention may be formulated into a preparation for oral or parenteral administration according to the route of administration as described above.

In the case of preparations for oral administration, the compositions of the present invention may be formulated using methods known in the art as powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions and the like. Can be. For example, oral formulations can be obtained by tablets or dragees by combining the active ingredients with solid excipients and then grinding them, adding suitable auxiliaries and processing them into granule mixtures. Examples of suitable excipients include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, including starch, corn starch, wheat starch, rice starch and potato starch, etc. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Furthermore, the pharmaceutical composition of the present invention may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, and the like.

Formulations for parenteral administration may be formulated by methods known in the art in the form of injections, creams, lotions, external ointments, oils, humectants, gels, aerosols and nasal inhalants. These formulations are described in Remington's Pharmaceutical Science, 19th ed., Mack Publishing Company, Easton, PA, 1995, a prescription generally known in all pharmaceutical chemistries.

 The total effective amount of the composition of the present invention may be administered to a patient in a single dose and may be administered by a fractionated treatment protocol which is administered in multiple doses for a long time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the extent of the disease. Preferably the preferred total dose of the pharmaceutical composition of the present invention may be about 0.01 μg to 10,000 mg, most preferably 0.1 μg to 500 mg per kg of patient body weight per day. However, the dosage of the pharmaceutical composition is determined in consideration of various factors such as the formulation method, route of administration and frequency of treatment, as well as various factors such as the patient's age, weight, health status, sex, severity of the disease, diet and excretion rate. In view of this, one of ordinary skill in the art will be able to determine the appropriate effective dosage of the compositions of the present invention. The pharmaceutical composition according to the present invention is not particularly limited to its formulation, route of administration and method of administration as long as the effect of the present invention is shown.

The compound of Formula 1 can regulate the migration of immune cells, and thus has a very significant effect in the prevention, improvement and treatment of diseases related to immune cell migration.

1A is a result of comparing the effects of collagen (Col), fibronectin (FN) and laminin (LN) on immune cell (monocyte / macrophage) migration by a transwell migration assay. Microscopic images of the migrating cells are shown.
Figure 1b is a graph showing the number of cells measured (quantitative) in the microscope image of Figure 1a.
FIG. 2A shows the results of comparing the effects of various laminin subtypes (LN111, LN211, LN221, LN411, LN421, LN511, LN521) on immune cell (monocyte / macrophage) migration by a transwell migration assay. As a microscopic image of the migrating cell is shown.
FIG. 2B is a graph showing the number of cells measured (quantified) in the microscopic image of FIG. 2A.
Figure 3 shows the results confirmed by Western blot KRS increase in monocytes / macrophage membrane by LN421 treatment.
Figure 4a shows a microscopic image of migrating cells as a result of comparing the effect of KRS expression on monocyte / macrophage migration specific to LN421 with a transwell cell migration assay.
Figure 4b is a graph showing the number of cells measured (quantitative) in the microscopic image of Figure 4a.
FIG. 5 shows that the KRS level was increased in the cell membrane region by treatment with LN421, which was significantly lowered by treatment with compounds having the structure of Formula 1 (typically BC-KI-00053). Indicates.
6A is a result of confirming that the migration of monocytes / macrophages is significantly inhibited in a concentration-dependent manner according to the treatment of compounds having the structure of Formula 1 (typically BC-KI-00053 compounds), Microscopic images of the migrating cells are shown.
FIG. 6B is a graph showing the number of cells measured (quantified) in the microscopic image of FIG. 6A.
FIG. 7A shows the results of fluorescence microscopy observation of monocytes, macrophages and Langerhans cells invasion following treatment with BC-KI-00053 compound in an acute inflammatory response (ear skin wound model). BC-KI-00053 100 mg / kg administration group, the lower part represents monocytes, macrophages and Langerhans cells, and the red part represents blood vessels labeled for CD31. skin wound location).
FIG. 7B is a quantitative representation of monocyte / macrophage infiltration at the periphery of the skin wound indicated by the blue circle in the fluorescence microscopy image of FIG. 7A.
Figure 8a is a schematic diagram of the triad (bile duct, hepatic artery, hepatic vein) occlusion procedure for the production of liver ischemia-reperfusion injury model.
Figure 8b shows the results of fluorescence microscopic observation of the infiltration rate of monocytes, macrophages and Cooper cells according to BC-KI-00053 compound treatment in liver ischemia-reperfusion injury model (the upper portion of the control group (vehicle administration group, the lower portion) Represents BC-KI-00053 100 mg / kg administration group, where the green areas represent monocytic cells, macrophages and Cooper cells, and the red areas represent blood vessels labeled for CD31).
FIG. 8c shows the quantitative degree of monocyte / macrophage infiltration in the fluorescence microscopy image of FIG. 8b quantitatively after time point after ischemia-reperfusion injury. The red bar is the result of quantification in the control group and the green bar in the BC-KI-00053 100 mg / kg group.
Figure 9a is a diagram showing the experimental method and schedule for the experiment to prepare a liver fibrosis animal model with CCl ₄ (carbon tetrachloride) and evaluate the therapeutic effect of the BC-KI-00053 compound.
FIG. 9B is a result of evaluating the effect of BC-KI-00053 in CCl ₄ (carbon tetrachloride) -induced liver fibrosis animal model. (In each figure, the upper part is an image of the liver surface, and the lower part is an image of the inside of the liver. The green part shows collagen and the red part shows hepatocytes).
FIG. 10A shows the change of right ventricular end-systolic pressure (RVESP) by BC-KI-00053 compound administration in pulmonary arterial hypertension (PAH) model (MCT: monocrotaline treated pulmonary arterial hypertension (PAH) model, Tx25mpk) : BC-KI-00053 25 mg / kg in PAH model, Tx50mpk: BC-KI-00053 50 mg / kg in PAH model).
FIG. 10B shows left ventricular end-systolic pressure (LVESP) change by BC-KI-00053 compound administration in pulmonary arterial hypertension (PAH) model (MCT: monocrotaline treated pulmonary arterial hypertension (PAH) model, Tx25mpk) : BC-KI-00053 25 mg / kg in PAH model, Tx50mpk: BC-KI-00053 50 mg / kg in PAH model).
Figure 10c is a result confirmed by IHC staining that immune cell migration and infiltration of lung tissue was reduced by BC-KI-00053 compound administration in pulmonary arterial hypertension (PAH) model.
FIG. 11A shows the base weight and weight change during the experimental period of vehicle treated and BC-KI-00053 treated groups in FHH rats with superimposed hypertension (numbers in parentheses used to calculate mean data in each group. Number of experimental animals, same as below).
Figure 11b is the result of measuring the change in MAP according to BC-KI-00053 treatment in FHH rats that are superimposed hypertension.
Figure 11c is the result of measuring the change in proteinuria (degree of protein excretion) in accordance with BC-KI-00053 treatment in FHH rats of superimposed hypertension (superimposed hypertension).
FIG. 11D shows the results of measuring plasma creatinine concentration change according to BC-KI-00053 treatment in FHH rats with superimposed hypertension.
FIG. 11E shows the results of evaluation of glomerular microscopy images (top image) and the degree of glomerulosclerosis of vehicle control group and BC-KI-00053 treatment group in FHH rats with superimposed hypertension (bottom) Graph) (the numbers inside the graph represent the number of images used to measure the actual results).
FIG. 11F quantifies the microscopic image (top image) and the degree of cortical fibrosis for cortical fibrosis of vehicle control group and BC-KI-00053 treatment group in FHH rats with superimposed hypertension. (Bottom graph) (the number inside the graph represents the number of images used to measure the actual result).
FIG. 11G shows the results of quantifying the microscopic image (top image) and the degree of water fibrosis of water fibrosis in vehicle control group and BC-KI-00053 treatment group in FHH rats with superimposed hypertension. (Bottom graph) (the number inside the graph represents the number of images used to measure the actual result).
FIG. 11H shows microscopic images (top image, right ventricular insertion point) and degree of cardiac fibrosis in vehicle control group and BC-KI-00053 treatment group in FHH rats with superimposed hypertension. The result shown by quantification is shown (bottom graph).
FIG. 11I shows IHC staining that immune cell migration and infiltration of kidney tissues were reduced by BC-KI-00053 compound administration in FHH rats with superimposed hypertension.
12A shows the base weight and duration of vehicle and BC-KI-00053 treated groups in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis and renal interstitial fibrosis through high salt (HS) diet. (Weights in parentheses represent the number of experimental animals used to calculate the mean data in each group. The same below).
Figure 12b is the result of measuring the change in MAP according to BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats induced high blood pressure through high salt (HS) diet.
FIG. 12C shows changes in proteinuria (degree of urinary protein excretion) following BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced proteinuria, glomerulosclerosis and renal interstitial fibrosis through high salt (HS) diet. One result.
FIG. 12D shows the results of measuring plasma creatinine concentration change according to BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis, and renal interstitial fibrosis through high salt (HS) diet. Indicates.
FIG. 12E shows vehicle control and BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis and renal interstitial fibrosis via high salt (HS) diet. Shows the result of evaluation of glomerular microscopy image (top image) and the degree of glomerulosclerosis (bottom graph) (the number inside the graph represents the number of images used to measure the actual result).
FIG. 12F shows vehicle control and BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis and renal interstitial fibrosis through high salt (HS) diet. Microscopic image (top image) and cortical fibrosis degree of quantification of the cortical fibrosis of the results are shown (bottom graph) (the number inside the graph represents the number of images used to measure the actual result).
FIG. 12G shows vehicle control and BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis and renal interstitial fibrosis through high salt (HS) diet. Microscopic image (top image) and water fibrosis degree for quantification of water are shown (bottom graph) (numbers in the graph indicate the number of images used to measure actual results).
FIG. 12H shows vehicle control and BC-KI-00053 treatment in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis and renal interstitial fibrosis via high salt (HS) diet. Microscopic image (top image, right ventricular insertion point) and cardiac fibrosis degree of quantification of cardiac fibrosis are shown (bottom graph).
12i shows immune cell migration and infiltration of kidney tissue by BC-KI-00053 compound administration in Dahl salt-sensitive (SS) rats that induced hypertension, proteinuria, glomerulosclerosis, and renal interstitial fibrosis via a high salt (HS) diet. This result was confirmed by IHC staining.
FIG. 13 shows the results of evaluating the infiltration of leukocytes and the reduction of fibrosis in the kidney when the control material or BC-KI-00053 compound was treated in an Alport syndrome animal model. CD45, a marker of leukocyte infiltration, was stained green, and collagen I, a marker of fibrosis, was stained red.

Hereinafter, the present invention will be described in detail.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1>

Identification of the role of laminin signaling in immune cell migration and infiltration

Among the extracellular matrix (ECM), which ones promote the migration and infiltration of monocytes / macrophages. Transwell migration assay was performed using collagen (collagen, Col), fibronectin, and laminin as extracellular matrix, and specific experimental methods are as follows. Transwells (Corning, # 3421-5mm) were coated with gelatin (0.5 mg / ml) and RAW 264.7 cells (1 × 10 ⁵ cells / well) were seeded into the top chamber. Serum Free DMEM (500 μl) containing 10 μg / ml of laminin (laminin mixture, Biolamina), Fibronectin or Collagen, respectively, was placed in the bottom chamber. After 24 hours, the cells were fixed with 30% of 70% methanol and stained with 50% Hematoxylin for 30 min. After removing non-migrating cells from the top of the membrane with a cotton swab, the membrane was taken and mounted on the slide. The migrating cells on the underside of the membrane were observed and quantified under a high magnification microscope.

As shown in Figure 1a and Figure 1b, it was confirmed that laminin of the various extracellular matrix most strongly promotes the migration of monocytes / macrophages. In other words, it was confirmed that the migration of monocytes / macrophages responded most strongly to laminin (LN) signals in the extracellular matrix (ECM).

<Example 2>

Immune Cell Migration and Invasion Effects by Laminin Subtype

In immune cell migration and infiltration, the effects of laminin subtypes were evaluated. A transwell migration assay was performed in the same manner as in Example 1 using LN111, LN211, LN221, LN411, LN421, LN511, and LN521 as various laminin subtype proteins (purchased from Biolamina). It was. The specific sequence of the laminin subtypes is according to the chain forming each laminin subtype, α4 chain of SEQ ID NO: 4, α2 chain of SEQ ID NO: 10, α5 chain of SEQ ID NO: 11, β2 chain of SEQ ID NO: 6, β1 chain of SEQ ID NO: 12 And the γ1 chain of SEQ ID NO: 8.

As shown in FIG. 2a and FIG. 2b, it was confirmed that monocytes / macrophages migrate in response to α4β2γ1 subtype (LN421) among laminin. That is, it was confirmed that the migration of monocytes / macrophages is specific for LN421 among laminin.

<Example 3>

Membrane migration of KRS due to laminin treatment in immune cells

After dispensing RAW 264.7 cells (2 x 10 ⁶ cells) in 100 pie plate and incubated for 18hr, LN421 1μg / ml treatment in serum free-DMEM media and harvested at 0h, 12h, 24h. RAW 264.7 cell proteins were separated into cytosol and membrane fractions using the ProteoExtract® Subcellular Proteome Extraction Kit (Calbiotech, cat # 539790). The obtained protein was electrophoresed and transferred to PVDF membrane (Milipore) and blocked with 3% skim milk. KRS was then detected by Western blot. Specifically, KRS polyclonal antibody (rabbit, Neomics, Co. Ltd. # NMS-01-0005) was added to the antibody for 1 hour. Unbound antibody was removed and reacted by the addition of anti-rabbit secondary antibody (ThermoFisher Scientific, # 31460). After reacting the secondary antibody, the film was photosensitive in the dark room using ECL reagent as a substrate. The photosensitized bands were compared to standard molecular markers to identify the bands corresponding to the size of the KRS. Antibodies against Na + / K + ATPase (Abcam, ab76020) and tubulin (Santa cruz SC-5286) were used to identify plasma membrane and cytosol markers, respectively.

As shown in FIG. 3, when the LN421 was treated to monocytes / macrophages, it was confirmed that the amount of KRS detection was increased in the cell membrane region as compared to a partial decrease in the amount of KRS detection in the cytosol region. These results suggest that KRS, which is expressed in the cells of monocytes / macrophages and is generally present in the cytoplasmic region, migrates to the cell membrane region by LN421 treatment. It is considered to be an important pathology for diseases related to immune cell migration and invasion.

<Example 4>

Effect of KRS on LN421-dependent immune cell migration and infiltration

To determine whether KRS influences the monocyte / macrophage migration specific to LN421, LN421 was treated with each of macrophages transformed to overexpress KRS and macrophages transformed to inhibit KRS expression, and transwell cells A transwell migration assay was performed. As a control, leucyl-tRNA synthetase (LRS, SEQ ID NO: 3), a KRS-like protein, was used.

Specifically, KRS or LRS overexpressing macrophages were constructed as follows: KRS-Myc and LRS-Myc inserted in pCDNA3 were transfected (48hrs) using Turbofect (Thermo Fisher Scientific) on Raw 264.7 cells, respectively. Cells transfected with Ev (empty vector, pCDNA3) -Myc were prepared as negative controls.

KRS or LRS expression-inhibiting macrophages were constructed as follows: Si-KRS (SEQ ID NO: 13) and Si-LRS (SEQ ID NO: 14) were respectively transfected with Lipofectamin (Thermo Fisher Scientific) to Raw 264.7 cells. 72hrs). As a negative control, cells transfected with si-control (si-RNA duplex with medium GC content (Invitrogen, Cat No. 12935-300)) were prepared.

The overexpression or suppression of expression of KRS or LRS in the transformed cells thus produced was confirmed by performing a western blot for each protein (data not shown).

For each of the transformed macrophages, a transwell migration assay was performed in the same manner as in Example 1 using 1 μg / ml of Laminin 421.

As shown in FIGS. 4A and 4B, KRS overexpression effectively increased LN421-specific monocyte / macrophage migration, whereas the reduction of KRS expression using si-RNA effectively prevented LN421-specific monocyte / macrophage migration. Reduced. In contrast, the expression of leucyl-tRNA synthetase (LRS), a KRS-like protein, did not affect monocyte / macrophage migration. This suggests that LN421-dependent migration of monocytes / macrophages is strongly influenced by KRS expression.

Example 5

Screening of Compounds That Inhibit the Migration of Immune Cells Without Side Effects_Inhibition of Movement of KRS to Cell Membranes

From the results of <Example 3> and <Example 4>, not only the expression level of KRS but also the intracellular behavior of KRS significantly influences LN421 dependent migration of monocytes / macrophages. The specific increase of KRS in the immune cell membrane region is considered to be an important pathology for diseases related to immune cell migration and invasion. Therefore, the aim of this study was to verify that inhibition of KRS could be one of the therapeutic strategies for diseases related to immune cell migration and invasion. On the other hand, KRS is an organ necessary for synthesizing proteins in cells under normal conditions. Therefore, simply increasing or decreasing the amount of KRS is likely to be inappropriate as a practical treatment strategy due to concerns about side effects on normal functioning of the body. Thus, the present inventors examined whether a specific known compound that affects the activity, intracellular kinetics and expression in various aspects of KRS can specifically inhibit migration to monocytes / macrophages without side effects.

As a result, the inventors confirmed that the compounds having the core structure of Formula 1 inhibit the migration of KRS to the cell membrane, thereby inhibiting LN431 specific monocyte / macrophage migration. This confirmation was specifically performed by the following in vitro experiments. Compounds represented by Formula 1 in the present invention were prepared and prepared according to the literature of the Republic of Korea Patent Publication 10-2018-0006167.

To determine whether KRS retains its inhibitory effect on the cell membranes of various KRS inhibitors, RAW 264.7 cells (2 x 10 ⁶ cells) were dispensed on 100 pie plates and incubated for 18 hours, and then serum-DMEM media was used. 12 min of incubation with laminin 421 1 μg / ml and 100 nM of each of the various KRS inhibitors. After harvesting, RAW 264.7 cell proteins were separated into cytosol and membrane fractions using ProteoExtract® Subcellular Proteome Extraction Kit (Calbiotech, cat # 539790). The obtained protein was electrophoresed and transferred to PVDF membrane (Milipore) and blocked with 3% skim milk. Since KRS was detected by Western blot, the specific method was performed in the same manner as in Example 3.

Thus, the substance identified to inhibit the migration of KRS to the cell membrane was treated with LN421-treated macrophages, and a transwell migration assay was performed. Through this, it was confirmed whether LN421-specific monocytes / macrophages have a migration inhibitory effect by inhibition of migration of KRS to the cell membrane. Specifically, the transwell (Corning, # 3421-5mm) was coated with gelatin (0.5mg / ml), and RAW 264.7 cells (1 × 10 ⁵ cells / well) were seeded into the top chamber. 500 μl of Serum Free DMEM containing 1 μg / ml of Laminin 421 (LN421, Biolamina) was placed in the bottom chamber. Thereafter, DMSO, or KRS inhibitor compound (in DMSO) was treated at various concentrations (30 nM, 100 nM, 300 nM, 1 μM, 3 μM, respectively) in the upper chamber. After 24 hours, the cells were fixed with 30% of 70% methanol and stained with 50% Hematoxylin for 30 min. After removing non-migrating cells from the top of the membrane with a cotton swab, the membrane was taken and mounted on the slide. The migrating cells on the underside of the membrane were observed and quantified under a high magnification microscope.

5 and 6 show the BC-KI-00053 compound (4-({(7-fluorobenzo [d] thiazol-2-yl)) having a particularly good effect among the compounds having the core structure of Chemical Formula 1. [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid) is representatively shown. As shown in FIG. 5, it was confirmed that the level of KRS in the cell membrane region by LN421 treatment was significantly lowered by BC-KI-00053. This means that the level of KRS migrated to the cell membrane of monocytes / macrophages by laminin (LN421) was reduced.

6A and 6B, it was confirmed that monocyte / macrophage migration was significantly inhibited depending on the concentration of BC-KI-00053 (inhibition of migration of KRS to the cell membrane).

In the following in vivo experiments on immune cell migration-related diseases, experiments were performed on behalf of the BC-KI-00053 compound.

<Example 6>

in vivo Effect of KRS Cell Membrane Inhibitor on Monocyte / Macrophage Infiltration in Acute Inflammatory Responses

<6-1> ear skin wound model

To investigate the effect of KRS membrane migration inhibitors on monocyte infiltration in acute inflammatory responses, ear skin wound models were constructed using CX3CR1-GFP mice (Stock no. # 005582, Jackson Laboratory (Bar Harbor, USA)). Monocytes, macrophages and langerhans cells appear green in CX3R1-GFP mice. Mice were administered orally with vehicle or BC-KI-00053 (100 mg / kg, dissolved in vehicle, once daily) for a total of 4 days prior to imaging 2 days (D-2, D-1, D-0, D + 1). Corn vehicle: Polyethylene glycol 400: Tween 80: Methyl cellulose (1%) = 20: 30: 1: 49 was used as a vehicle. A 31 G syringe was used to puncture the skin of the ear (time D-0) to induce an acute inflammatory response. Blood vessels were labeled using an anti-CD31 antibody bound to Alexa Flour 555 (identified in red). Confocal microscopy was used as the imaging equipment.

As shown in FIGS. 7A and 7B, in the vehicle-treated control group, monocytes and macrophages gathered around the wounded area (ear hole area) indicated by the white circle, and it was confirmed that they were infiltrated over a wide range to a high level. (Blue circle). In contrast, monocyte and macrophage infiltration were significantly reduced in mice administered BC-KI-00053. Meanwhile, in FIG. 7A and FIG. 7B, the green dots appearing to be scattered around the general tissue rather than the main lesion (blue circle) are Langerhans cells, which are resident macrophages, and Langerhans cells in both the control group and the BC-KI-00053 treatment group. It was judged that only migratory macrophage migration was inhibited by the treatment of BC-KI-00053 because it did not affect the number of. From the above results, it was confirmed that the migration and infiltration of immune cells due to the acute inflammatory response were inhibited by the administration of KRS membrane inhibitors (in particular, BC-KI-00053), which means that the compounds of the present invention were pro-inflammatory cytokines. By inhibiting the excessive movement of the immune cells that secrete caine means that it can have a prophylactic or therapeutic effect against inflammatory diseases.

<6-2> Liver Ischemia-Reperfusion Injury Model

Liver Ischemia-Reperfusion Injury Model was constructed using CX3CR1-GFP mice to investigate the effect of KRS membrane migration inhibitors on monocyte infiltration in ischemic immune responses. Monocytes, macrophages and kupffer cells appear green in CX3CR1-GFP mice. Mice were administered orally with vehicle or BC-KI-00053 (100 mg / kg, dissolved in vehicle, once daily) for 3 days from 2 days before imaging (D-2, D-1, D-0). Corn oil (Polyethylene glycol 400: Tween 80: Methyl cellulose (1%) = 20: 30: 1: 49) was used as a vehicle. On day 3 of oral administration (D-0), a 6-0 suture was used, and triad (bile duct, hepatic artery, hepatic vein) occlusion was performed as shown in FIG. 8A. Triad occlusion was performed for 30 minutes to induce acute inflammation, and 3g of Eppendorf tubes were suspended from both ends of the suture. The sutures were removed and the ischemic inflammation site was observed immediately after reperfusion (0h) and 24h. At this time, blood vessels were labeled with Alexa Flour 555-conjugated anti-CD31 antibody for repeated imaging. ). Two-photon microscope was used as the imaging equipment.

As shown in FIG. 8B and FIG. 8C, in the vehicle-treated control mouse, a large number of monocytes / macrophages were infiltrated into the wound site (blocking site) after 24 hours of reperfusion. Confirmed. In the experimental group treated with BC-KI-00053, monocyte and macrophage invasion was significantly reduced. In FIG. 8C, the red bars are the results of quantification of the control group and the green bars of the BC-KI-00053 100 mg / kg administration group, respectively. Meanwhile, in FIG. 8B, the very bright green cells appearing to be scattered around the general tissue mainly seen at reperfusion 0h are resident macrophage kupffer cells, and both control and BC-KI-00053 treated kupffer cells after 24h reperfusion. It was judged that only migratory macrophage migration was inhibited by the treatment of BC-KI-00053 because it did not affect the number of. In other words, KRS cell membrane migration inhibitors (particularly BC-KI-00053) have an excellent effect of inhibiting only monocyte / macrophage infiltration that migrates to ischemia-induced liver.

<Example 7>

In vivo  Pharmacological effects of KRS cell membrane migration inhibitors on liver fibrosis

Hepatocytes appear red in Actin-DsRed (Stock no. # 006051, Jackson Laboratory (Bar Harbor, USA)) mice. In order to induce liver fibrosis of the mouse, CCl ₄ (carbon tetrachloride) was dissolved in corn oil, and twice a week at a concentration of 20%, for 6 weeks, and intraperitoneal injection was performed. Three weeks after the start of CCl ₄ administration, the vehicle and the BC-KI-00053 (100 mg / kg) were administered daily, orally, for three weeks. Corn vehicle: Polyethylene glycol 400: Tween 80: Methyl cellulose (1%) = 20: 30: 1: 49 was used as a vehicle. Animal groups were configured as shown in Table 1 below.

The degree of fibrosis of the liver surface and inside (area of 30 to 50 μm deep) was detected by the second harmonic generation (SHG) technique of intravial imaging (Excitation: 780 nm, Detection: 390 nm).

division Material handling status Experimental animals
head Control (One) Cone oil administration animal (normal animal) + control vehicle (vehicle) administration One (2) Cone oil administration animal (normal animal) + BC-KI-00053 administration One Experimental group (3) CCl ₄ liver fibrosis animal model + vehicle administration 2 (4) CCl ₄ Liver Fibrosis Animal Model + BC-KI-00053 Administration 3

No weight loss was seen in the group to which BC-KI-00053 was administered to normal animals (animal No. 2 in Table 1), and no other symptoms were caused in the liver. Therefore, BC-KI-00053 compound was considered to be harmless in vivo . The group administered the vehicle to the fibrotic animal model (animals in (1) in Table 1) died early due to the toxicity of CCl ₄ at 4 weeks after the start of the experiment. Specifically, as shown in FIG. 9A, one of the six weeks of the experiment died on day 24 and the other on day 32, and they showed significant weight loss. The early dead test group was used for comparison of data after all experiments were completed, using liver surface and internal fibrosis data obtained at 2 and 4 weeks, respectively, before they died.

In general, severe fibrosis is observed in a linear shape rather than in a curled appearance. As shown in FIG. 9B, a significant fibrosis was observed on the surface of the liver (top (0 μm) of FIG. 9B) after CCl ₄ administration for 2 or 4 weeks, especially in the group administered CCl ₄ for 4 weeks. The shape appeared more and the septum (portal-portal septa) was found that the severe fibrosis was progressed significantly. In addition, similar fibrosis was observed in the liver (bottom (30-48 μm) of FIG. 9B), and it was also confirmed that hepatocytes (necrosis) were at a severe level. On the other hand, in the animal group treated with BC-KI-00053 for three weeks during the six-week administration of CCl ₄ (animal group No. 4 in Table 1), fibrosis on the surface and inside of the liver was significantly reduced compared to the CCl ₄ two-week group. It was confirmed that the appearance of capsular collagen similar to that of normal subjects, and also significantly reduced hepatocyte necrosis. In FIG. 9B, green represents collagen (fibrosis) and red represents hepatocytes. These experimental results show that the KRS cell membrane migration inhibitor (in particular, BC-KI-00053) provided by the present invention has excellent fibrosis inhibitory ability.

<Example 8>

In vivo  Pulmonary arterial hypertension ( PAH For) KRS  Confirmation of Pharmacological Effect of Membrane Migration

Experimental Method

1. Preparation of pulmonary hypertension (PAH) model and administration of test substance

To induce PAH in 6-week-old female SD rats (Oriental Bio), 60 mg / kg of MCT (monocrotaline), a substance that causes pulmonary hypertension through selective injury of the pulmonary artery, was injected subcutaneously. Then divided into 4 groups (tested with 5 animals in each group), vehicle, sildenafil (25 mg / kg, once daily) in rats or BC-KI-00053 (25 or 50 mg / kg, dissolved in vehicle, Once daily) orally for 3 weeks. Corn vehicle: Polyethylene glycol 400: Tween 80: Methyl cellulose (1%) = 20: 30: 1: 49 was used as a vehicle.

2. Blood flow and blood pressure measurement

Three weeks later, rats were anesthetized with isoflurane, and blood flow and pressure were measured using an animal high precision pneumatic pressure measurement system (MPVS Cardiovascular Pressure and Volume system, model name: MPVS Ultra, manufacturer: Millar Instruments). Right ventricular systolic pressure (RVESP) and diastolic pressure, left ventricular systolic pressure and diastolic pressure were measured using a dedicated catheter (Mikro-Tip ^® ratpressure catheter, manufacturer: Millar Instruments). Cardiac output was measured using a perivascular blood flow probe (Transonic ^® Flowprobes, manufacturer: Millar Instruments), and the experimental technique was performed in the same manner as described in the literature: Pacher P, Nagayama T, Mukhopadhyay P, Batkai S, Kass DA. Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats. Nat Protoc 2008; 3 (9): 1422-34.

3. Immunohistochemistry (IHC)

IHC staining for CD68, a monocyte / macrophage marker, was performed using lung tissue from each experimental group. The collected lungs were fixed in PFA (paraformaldehyde) according to a conventional procedure, and then embedded in paraffin through water washing, dehydration, and transparent processes. Lung tissue paraffin blocks of rats were cut to a thickness of 6 μm and slides were prepared. Thereafter, staining was performed as follows. First, three times xylene treatment for 5 minutes, 2 minutes in 100% ethanol twice, 95% ethanol, 90% ethanol, 70% ethanol, DW for 2 minutes and washed with PBS for 5 minutes (2 times). After treatment with 0.3% H ₂ O ₂ (10 minutes), the samples were washed twice with PBS for 5 minutes. Soak in 0.01M citrate buffer of pH 6.0 using microwave for 3 minutes and 30 seconds, and then cooled for 10 seconds and cooled for 10 minutes at room temperature after repeated 10 seconds heating (antigen retrieval). Then washed three times with 5 minutes each PBS-T (0.03% Triton-X). After 30 minutes blocking (2% BSA & 2% goat serum in PBS) at 4 ℃. Anti-CD68 antibody (1: 200, Abcam, ab31630) was treated overnight at 4 ° C. After washing three times with PBS-T for 5 minutes, it was treated with polymer-HRP amti-mouse envision kit (DAKO) for 1 hour at 4 ℃. After washing three times with PBS-T for 5 minutes, 1ml of DAB substrate buffer and 20ul of DAB chromogen were mixed and treated with tissue. After 10 minutes, the cells were washed twice with 3rd DW for 2 minutes. The stained tissue was treated with Mayer's hematoxylin (Sigma) for 1 minute, and then treated twice for 2 minutes in order of 70% ethanol, 90% ethanol, 95% ethanol, and 100% ethanol. Finally, after xylene treatment 3 times for 5 minutes, the mounting was carried out using a mounting solution, and observed with an optical microscope.

[Experiment result]

Pulmonary hypertension causes the right ventricular pressure to rise due to narrowing of the pulmonary artery, resulting in right ventricular failure. In addition, if the reward mechanism is destroyed by persistent hypertension, right ventricular enlargement is followed by right ventricular enlargement. This results in compression of the left ventricle due to displacement of the ventricular septum and reduction of the left ventricular dilatation volume and cardiac output (Lee Woo-seok et al., Clinical characteristics and prognostic factors in patients with severe pulmonary hypertension, Korean Circulation J 2007; 37 : 265-270 ). As a result, pulmonary hypertension is primarily associated with the right ventricle but also with the function of the left ventricle.

As shown in FIG. 10A, it was confirmed that RVESP (right ventricular systolic pressure) was increased in the PAH animal model, whereas treatment with BC-KI-00053 significantly reduced RVESP in a concentration dependent manner. In particular, the effect of lowering RVESP in BC-KI-00053 50 mg / kg treatment group was similar to that of sildenafil, one of the standard treatments.

In addition, there was no decrease in the left ventricular end systolic pressure (LVESP) according to the treatment of BC-KI-00053, but rather, the LVESP was increased in the BC-KI-00053 50 mg / kg administration group as shown in FIG. 10B. I was not statistically significant. This is in contrast to the risk of lowering systemic blood pressure in sildenafil, which is used as a conventional treatment for pulmonary hypertension, causing not only pulmonary artery expansion but also systemic artery expansion. In other words, BC-KI-00053 showed a lower tendency to affect systemic artery pressure than sildenafil, and this effect is concerned with the risk of hypotension when sildenafil is administered in clinical settings. Considering this, it is considered to be an advantageous property of therapeutic agent. In addition, when pulmonary hypertension is severe, as right ventricular failure occurs, low cardiac output and systemic hypotension may be accompanied. In contrast, the treatment of BC-KI-00053 at higher concentrations may improve cardiac output and systemic blood pressure. If cardiac output and systemic blood pressure are lowered, the patient may complain of general weakness or dizziness. Therefore, improvement of cardiac output and systemic blood pressure may be expected to improve these symptoms.

Taken together, this suggests that the administration of KRS cell membrane migration inhibitors (especially BC-KI-00053) provided by the present invention not only exhibits PAH therapeutic effects and symptomatic relief, but also may present a relatively low risk of developing side effects of existing therapeutic drugs. Confirmed.

In addition, as shown in FIG. 10C, IHC staining for CD68, a monocyte / macrophage marker using lung tissue of each experimental group, showed that monocyte / macrophage infiltration occurs in the lungs of pulmonary arterial hypertension (PAH) mice. It confirmed that there was. In contrast, it was confirmed that the treatment of the KRS cell membrane migration inhibitor (in particular, BC-KI-00053) provided by the present invention clearly reduced lung tissue invasion of monocytes / macrophages, and this effect was previously effective for pulmonary hypertension. It was confirmed that it is significantly superior to sildenafil known to be.

Example 9

in vivo  For hypertension-induced proteinuria, glomerulosclerosis, kidney and heart fibrosis KRS  Confirmation of Pharmacological Effect of Membrane Migration

<9-1> superimposed hypertension FHH In the rat KRS  Effects of Membrane Migration Inhibitors on Hypertensive Kidney Injury, Heart Injury and Fibrosis Development

Experimental Method

Experiments were performed using male FHH rats 9-12 weeks of age. These animals were provided by the University of Mississippi Medical Center and approved by the American Association for Accreditation of Laboratory Animal Care (AAALAC). All protocols are approved by the Institutional Animal Care and Use Committee of the University of Mississippi Medical Center. The rats were freely fed food and water, and these rats were provided with a purified AIN-76 rodent feed containing 0.4% NaCl (Dyets, Bethlehem, PA) after weaning. Fawn-hooded hypertensive (FHH) rats are a genetic model of spontaneous hypertension associated with glomerular hyperfiltration and proteinuria. In order to promote glomerular damage in the rat, DOCA strips were implanted after single (one) kidney extraction.

Specifically, FHH rats were anesthetized with isoflurane and telemetry transmitters (model TA11PAC40, Data Sciences International, St. Paul, MN) were described in Williams, JM et al. Am J Physiol Regul Implants as described in Integr Comp Physiol (2012). In brief, the operation was performed under 2-3% isoflurane-O ₂ , and the catheter of the device was inserted into the left femoral artery and guided upstream to the aorta. The body of the telemetry unit was placed in the lateral cavity of the left leg and sutured with muscle tissue. The skin was then closed. To prevent infection, animals were given Baytril (10 mg / kg) and Rimadyl (5 mg / kg), a long-acting analgesic, to control surgical pain. After surgery, rats were housed in individual cages in a quiet air-conditioned room environment with a 12: 12-h contrast cycle and took a week to fully recover from surgery. Thereafter, the basic mean arterial blood pressure (MAP) and proteinuria were measured for 4 hours (10 am to 2 pm) before the rats were housed in the metabolic cage. Proteinuria was measured using the Bradford method and BSA (Bio-Rad Laboratories, Hercules, CA) as standard.

One week after the insertion of the transmitter, rats were treated with Wang, X. et al. Am J Physiol Uninephrectomized as described in Renal Physiol (2016). Briefly, rats were anesthetized with 2-3% isoflurane-O ₂ and the right flank was dissected in aseptic condition. The right kidney was gently lifted and threaded tightly around the renal vessels and ureters. Kidneys were extracted by cutting the renal vessels and distal ends of the ureters. The incision is closed with a continuous subcutaneous stitch, after which the skin is further closed. After the rat's right kidney was removed, DOCA pellets (200 mg, Innovative Research of America) were implanted subcutaneously in the neck area.

After single kidney extraction and DOCA transplant surgery, rats had recovery time for 3 days. The rats were then provided with distilled water, converted to water containing 1% NaCl, and randomly divided into two groups: Group 1 (n = 15) BC through gastrointestinal tract gavage. -KI-00053 (25 mg / kg daily) was treated; Group 2 (n = 15) was administered directly to the gastrointestinal tract in the same volume (2.5 ml / kg of vehicle) (corn oil, polyethylene glycol 400, Tween 80 and methylcellulose). Blood pressure and proteinuria were measured weekly for 3 weeks in the experimental group. At the end of the experiment, rats were anesthetized with isoflurane and blood samples were taken to measure creatinine levels. Rats were then flushed with 50 ml of 0.9% NaCl through aorta and perfused with 20 ml of 4% paraformaldehyde. Kidneys and hearts were collected for histological evaluation.

Paraffin sections prepared at 3 μm thickness were stained with Masson's trichrome to measure the degree of glomerular damage and renal interstitial fibrosis. Images were obtained using a Nikon Eclipse 55i microscope with Nikon DS-Fi1 color camera (Nikon, Melville, NY) and NIS-Elements D 3.0 software. The degree of glomerular damage was assessed by the experimenter from 0 to 4+ with no preconception on 30-40 glomeruli / section. 0 represents normal glomeruli, 1+ represents 1-25% loss, 2+ represents 26-50% loss, 3+ represents 51-75% loss, and 4+ represents capillaries in tufts It indicates that the loss is more than 75%. Cortical and medulla fibrosis were analyzed using NIS-Elements automated measurement software after thresholding to determine the percentage of images stained in blue. In addition, immunohistochemical staining (IHC) for CD68, a monocyte / macrophage marker for kidney tissue, was performed in the same manner as in Example 8.

Statistics: Each data is expressed as mean ± SEM. Comparison between groups was analyzed by two-tailed test. P value <0.05 was considered statistically significant.

[Experiment result]

There was no difference in base weight between the vehicle treatment group and the BC-KI-00053 treatment group (control 309.57 ± 4.14 g, experimental 304.7 ± 5.39 g, P> 0.05). Body weight was reduced by about 10% in vehicle or BC-KI-00053 treated rats during the study but there was no statistical difference between the two groups (FIG. 11A).

MAP data measured via telemetry in control and experimental FHH rats are shown in FIG. 11B. There was no difference in base MAP between the two groups (control 120.50 ± 0.91 mmHg, experimental group 120.1 ± 0.62 mmHg, P> 0.05). MAP increased rapidly in both groups after single resection and conversion to 1% NaCl feed with DOCA pellet insertion. Vehicle treatment group had a larger gap in MAP than BC-KI-00053 treatment group. After one week of treatment, the MAP of the BC-KI-00053 treatment group was statistically lower than that of the vehicle treatment group (control group 184.34 ± 2.46 mmHg, experimental group 174.4 ± 3.83 mmHg, P <0.05). After 2 weeks of treatment, the MAP results in the vehicle treatment group appeared to be relatively stable compared to the results of the first week. MAP in the BC-KI-00053 group was transient but decreased further, with significant differences between the two groups (control 184.22 ± 4.21 mmHg, experimental group 168.8 ± 3.74 mmHg, P <0.05). Three weeks later, the mean MAP difference between the two groups widened (control 195.30 ± 3.68 mmHg, experimental group 176.9 ± 5.83 mmHg, P <0.05).

Data for proteinuria in control and experimental FHH rats are shown in FIG. 11C. There was no difference in baseline proteinuria between the two groups (control 52.75 ± 6.99 mg / day, experimental 51.0 ± 4.9 mg mg / day, P> 0.05). After a single resection, the conversion to 1% NaCl feed with DOCA pellet insertion increased the proteinuria in both groups. After two weeks of treatment, the proteinuria in the BC-KI-00053 treated group was statistically lower compared to the vehicle treated group (control 472.99 ± 53.81 mg / day, experimental group 285.5 ± 47.48 mg / day, P <0.05). Continued to end of study (control 675.61 ± 49.91 mg / day, control 433.1 ± 60.59 mg / day, P <0.05)

Data for plasma creatinine concentrations in control and experimental FHH rats are shown in FIG. 11D. Plasma creatinine concentration in vehicle treated group was significantly higher than BC-KI-00053 treated group (control group 0.65 ± 0.04 mg / dL, experimental group 0.48 ± 0.02 mg / dL, P <0.05).

Conversion to 1% NaCl feed with DOCA pellet insertion after single resection in FHH rats had a significant effect on morphologically glomerular and coronary injury (FIG. 11E, FIG. 11F, FIG. 11G). The mean glomerular injury score (score) showed that the degree of injury was significantly reduced in the rats treated with BC-KI-00053 (control 3.16 ± 0.04, experimental 1.49 ± 0.05, P <0.05). In addition, the fibrosis was significantly decreased in the BC-KI-00053 treatment group, whereas the fibrosis was significantly increased in the vehicle treatment group. Specifically, BC-KI-00053-treated rats showed significantly less cortical fibrosis (control group 19.46 ± 1.18%, experimental group 5.79 ± 0.48%, P <0.05), and linear vessel capillaries in renal medulla fibrosis and coronary injury. Vascular loss was significantly reduced (control 17.69 ± 1.07%, experimental 7.40 ± 0.56%, P <0.05).

As seen in the section sample stained with Sirius red (FIG. 11H), control rats showed significant cardiac fibrosis, especially at the right ventricular insertion point. In contrast, the rate of cardiac fibrosis was significantly decreased in the rats treated with BC-KI-00053 (control group 31.97 ± 2.62%, experimental group 9.14 ± 2.18%, P <0.05).

In addition, as a result of confirming the degree of macrophage invasion, as shown in FIG. 11I, IHC staining for CD68, which is a monocyte / macrophage marker using renal tissue, showed monocyte / macrophage infiltration in the renal tissue of the control group (vehicle treatment). It was confirmed that this happened at a high level. In contrast, it was confirmed that the treatment of the KRS cell membrane migration inhibitor (especially BC-KI-00053) provided in the present invention significantly reduced renal tissue infiltration of monocytes / macrophages.

<9-2> SS In the rat KRS  Effects of Membrane Migration Inhibitors on Hypertensive Kidney Injury, Heart Injury and Fibrosis Development

Experiments were performed using male SS rats 9-12 weeks of age. These animals were provided by the University of Mississippi Medical Center and approved by the American Association for Accreditation of Laboratory Animal Care (AAALAC). All protocols are approved by the Institutional Animal Care and Use Committee of the University of Mississippi Medical Center. The rats were freely fed food and water, and these rats were provided with a purified AIN-76 rodent feed containing 0.4% NaCl (Dyets, Bethlehem, PA) after weaning. Dahl salt-sensitive (SS) rats are animal models that rapidly induce high hypertension, proteinuria, glomerulosclerosis, and renal interstitial fibrosis in high salt (HS) diets.

SS rats were anesthetized with isoflurane and telemetry transmitters (model TA11PAC40, Data Sciences International, St. Paul, MN) were sterilely implanted in the same manner as described above. After surgery, rats were housed in individual cages in a quiet air-conditioned room environment with a 12: 12-h contrast cycle and took a week to fully recover from surgery. Then, the primary mean arterial blood pressure (MAP) was measured before the rats were housed in the metabolic cage to measure urine protein excretion. Proteinuria was measured using the Bradford method and BSA (Bio-Rad Laboratories, Hercules, CA) as standard.

Rats were then divided into two randomized experimental groups: Group 1 (n = 15) was treated with BC-KI-00053 (25 mg / kg daily) via gastrointestinal tract gavage; Group 2 (n = 15) was directly administered to the gastrointestinal tract in the same volume (2.5 ml / kg per day) of vehicles (corn oil, polyethylene glycol 400, Tween 80 and methylcellulose). Simultaneously with the material treatment, the feed was changed to HS feed containing 8% NaCl (Dyets, Bethlehem, PA) and blood pressure and proteinuria were measured at 7, 14 and 21 days after HS feed. At the end of the experiment, rats were anesthetized with isoflurane and blood samples were taken to measure creatinine levels. Rats were then flushed with 50 ml of 0.9% NaCl through aorta and perfused with 20 ml of 4% paraformaldehyde. Kidneys and hearts were collected for histological evaluation.

Paraffin sections were fabricated and evaluations of the degree of glomerular damage, cortex and medulla fibrosis were performed as described above. In addition, immunohistochemical staining (IHC) for CD68, a monocyte / macrophage marker for kidney tissue, was performed in the same manner as in Example 8.

Statistics: Each data is expressed as mean ± SEM. Comparison between groups was analyzed by two-tailed test. P value <0.05 was considered statistically significant.

[Experiment result]

There was no difference in base weight between the vehicle and BC-KI-00053 treated groups (control 337.92 ± 9.86 g, experimental 350.13 ± 9.173 g, P> 0.05). Body weights were maintained or increased slightly in Vehicle or BC-KI-00053 treated rats, but there was no statistical difference between the two groups for the entire study period (FIG. 12A).

MAP data measured via telemetry in control and experimental SS rats are shown in FIG. 12B. There was no difference in base MAP between the two groups (control group 122.13 ± 2.31 mmHg, experimental group 123.45 ± 2.36 mmHg, P> 0.05). The change in feed from SS rats to HS feed continued to increase MAP in both groups. Vehicle treatment group had a larger gap in MAP than BC-KI-00053 treatment group. After 2 weeks of treatment, the MAP of BC-KI-00053 treatment group was statistically decreased than the vehicle treatment group (control group 178.51 ± 3.71 mmHg, experimental group 164.43 ± 3.00 mmHg, P <0.05), and this effect was terminated. Until control (control 201.65 ± 2.54 mmHg, 178.48 ± 3.49 mmHg, P <0.05).

Data for proteinuria in control and experimental SS rats are shown in FIG. 12C. There was no difference in baseline proteinuria between the two groups (control 133.82 ± 10.50 mg / day, experimental group 113.27 ± 8.06 mg / day, P> 0.05). The conversion of SS rats into HS diet led to a sharp increase in proteinuria in both groups. In particular, the vehicle treatment group was confirmed to increase the proteinuria significantly higher than the BC-KI-00053 treatment group. After 1 week of treatment, the proteinuria in the BC-KI-00053 treated group was statistically lower than the vehicle treated group (control 469.08 ± 24.82 mg / day, experimental group 302.86 ± 29.76 mg / day, P <0.05). . After two weeks of treatment, the proteinuria levels in the BC-KI-00053 and vehicle treatment groups were still clearly different (control 675.61 ± 59.67 mg / day, experimental group 510.64 ± 42.42 mg / day, P <0.05). Continued to end of study (control 752.97 ± 57.80 mg / day, control 524.55 ± 44.70 mg / day, P <0.05)

Data for plasma creatinine concentrations in control and experimental SS rats are shown in FIG. 12D. Plasma creatinine concentration in vehicle treated group was significantly higher than BC-KI-00053 treated group (control group 0.60 ± 0.02 mg / dL, experimental group 0.55 ± 0.01 mg / dL, P <0.05).

Providing an HS diet had a significant effect on morphologically glomerular and coronary injury in SS rats (FIG. 12E, FIG. 12F, FIG. 12G). The mean glomerular injury score (score) showed that the degree of injury was significantly reduced in the rats treated with BC-KI-00053 (control 2.82 ± 0.05, experimental group 1.34 ± 0.04, P <0.05). In addition, the fibrosis was significantly decreased in the BC-KI-00053 treatment group, whereas the fibrosis was significantly increased in the vehicle treatment group. Specifically, BC-KI-00053-treated rats showed significantly less fibrosis of the cortex (control group 19.48 ± 0.96%, experimental group 6.47 ± 0.46 %%, P <0.05), and the straight blood vessels in renal medulla fibrosis and coronary injury. Capillary loss was significantly reduced (control group 23.49 ± 0.99%, experimental group 12.33 ± 0.78%, P <0.05).

As seen in section samples stained with Sirius red (FIG. 12H), control rats showed significant cardiac fibrosis, especially at the right ventricular insertion point. In rats treated with BC-KI-00053, the rate of cardiac fibrosis was significantly reduced (control group 18.60 ± 0.93%, experimental group 6.63 ± 0.94%, P <0.05).

In addition, as a result of confirming the degree of macrophage invasion, as shown in FIG. 12i, IHC staining for CD68, which is a monocyte / macrophage marker using kidney tissue, was found to be monocyte / macrophage infiltration in the kidney tissue of the control group (vehicle treatment). It was confirmed that this happened at a high level. In contrast, it was confirmed that the treatment of the KRS cell membrane migration inhibitor (especially BC-KI-00053) provided in the present invention significantly reduced renal tissue infiltration of monocytes / macrophages.

<Example 10>

KRS  Cell membrane migration inhibitors  in vivo Alport  Effects on Kidney Fibrosis and Immune Cell Infiltration in Syndrome Animal Models

The experiment was conducted using 129 Sv / J mice (Boys town hospital). The experimental animals were treated with (i) 129 Sv / J wild-type mouse control (0.5% methylcellulose suspension), (ii) 129 Sv / J alport mice (COL4A3 knockout mouse. Cosgrove D et al., Genes Dev. 1996 Dec 1 10 (23): 2981-92) to control (0.5% methylcellulose suspension) administration group and (iii) BC-KI-00053 administration group to 129 Sv / J alport mice. Each dose group consists of two birds. BC-KI-00053 group was dissolved in 0.5% methylcellulose suspension and orally administered at a concentration of 100 mg / kg, and kidney fibrosis and immune cell infiltration were evaluated. Each animal group was treated with a control or drug once a day from 3 weeks of age for a total of 4 weeks. After 4 weeks of drug treatment, renal paraffin sections were stained with collagen I (marker of fibrosis) and CD45 to observe the extent of leukocyte infiltration. Fibrosis and infiltration confirmation were performed in the same manner as used in the above examples.

As can be seen in Figure 13, the control group (vehicle, 0.5% methyl cellulose) was administered to the Alport mice, it was confirmed that the leukocyte infiltration and fibrosis significantly progressed in the kidney. In contrast, it was confirmed that the leukocyte infiltration was reduced and fibrosis was reduced in normal (wild-type mouse-controlled group) level in the Alport mouse kidney treated with BC-KI-00053.

As described above, the present invention relates to a novel use of benzo [d] thiazole derivatives having a specific structure disclosed in the present invention, and more particularly, to benzo [d] thiazole having a structure of Formula 1 The present invention relates to a prophylactic or therapeutic use of a disease related immune cell migration. The compound of formula 1 can regulate the migration of immune cells, and thus has a very significant effect in the prevention, improvement and treatment of diseases related to immune cell migration, and thus has high industrial applicability in the pharmaceutical industry.

<110> Medicinal Bioconvergence Research Center          Korea Advanced Institute of Science and Technology          SEOUL NATIONAL UNIVERSITY HOSPITAL <120> Pharmaceutical composition for preventing or treating immunocyte          migration-related diseases comprising benzo [d] thiazole          derivatives <130> NP17-0123 <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 597 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of KRS (Homo sapiens) <400> 1 Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro   1 5 10 15 Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys              20 25 30 Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu          35 40 45 Ser Gln Ala Thr Ala Ala Ala Thr Asn His Thr Thr Asp Asn Gly Val      50 55 60 Gly Pro Glu Glu Glu Ser Val Asp Pro Asn Gln Tyr Tyr Lys Ile Arg  65 70 75 80 Ser Gln Ala Ile His Gln Leu Lys Val Asn Gly Glu Asp Pro Tyr Pro                  85 90 95 His Lys Phe His Val Asp Ile Ser Leu Thr Asp Phe Ile Gln Lys Tyr             100 105 110 Ser His Leu Gln Pro Gly Asp His Leu Thr Asp Ile Thr Leu Lys Val         115 120 125 Ala Gly Arg Ile His Ala Lys Arg Ala Ser Gly Gly Lys Leu Ile Phe     130 135 140 Tyr Asp Leu Arg Gly Glu Gly Val Lys Leu Gln Val Met Ala Asn Ser 145 150 155 160 Arg Asn Tyr Lys Ser Glu Glu Glu Phe Ile His Ile Asn Asn Lys Leu                 165 170 175 Arg Arg Gly Asp Ile Ile Gly Val Gln Gly Asn Pro Gly Lys Thr Lys             180 185 190 Lys Gly Glu Leu Ser Ile Ile Pro Tyr Glu Ile Thr Leu Leu Ser Pro         195 200 205 Cys Leu His Met Leu Pro His Leu His Phe Gly Leu Lys Asp Lys Glu     210 215 220 Thr Arg Tyr Arg Gln Arg Tyr Leu Asp Leu Ile Leu Asn Asp Phe Val 225 230 235 240 Arg Gln Lys Phe Ile Ile Arg Ser Lys Ile Ile Thr Tyr Ile Arg Ser                 245 250 255 Phe Leu Asp Glu Leu Gly Phe Leu Glu Ile Glu Thr Pro Met Met Asn             260 265 270 Ile Ile Pro Gly Gly Ala Val Ala Lys Pro Phe Ile Thr Tyr His Asn         275 280 285 Glu Leu Asp Met Asn Leu Tyr Met Arg Ile Ala Pro Glu Leu Tyr His     290 295 300 Lys Met Leu Val Val Gly Gly Ile Asp Arg Val Tyr Glu Ile Gly Arg 305 310 315 320 Gln Phe Arg Asn Glu Gly Ile Asp Leu Thr His Asn Pro Glu Phe Thr                 325 330 335 Thr Cys Glu Phe Tyr Met Ala Tyr Ala Asp Tyr His Asp Leu Met Glu             340 345 350 Ile Thr Glu Lys Met Val Ser Gly Met Val Lys His Ile Thr Gly Ser         355 360 365 Tyr Lys Val Thr Tyr His Pro Asp Gly Pro Glu Gly Gln Ala Tyr Asp     370 375 380 Val Asp Phe Thr Pro Pro Phe Arg Arg Ile Asn Met Val Glu Glu Leu 385 390 395 400 Glu Lys Ala Leu Gly Met Lys Leu Pro Glu Thr Asn Leu Phe Glu Thr                 405 410 415 Glu Glu Thr Arg Lys Ile Leu Asp Asp Ile Cys Val Ala Lys Ala Val             420 425 430 Glu Cys Pro Pro Pro Arg Thr Thr Ala Arg Leu Leu Asp Lys Leu Val         435 440 445 Gly Glu Phe Leu Glu Val Thr Cys Ile Asn Pro Thr Phe Ile Cys Asp     450 455 460 His Pro Gln Ile Met Ser Pro Leu Ala Lys Trp His Arg Ser Lys Glu 465 470 475 480 Gly Leu Thr Glu Arg Phe Glu Leu Phe Val Met Lys Lys Glu Ile Cys                 485 490 495 Asn Ala Tyr Thr Glu Leu Asn Asp Pro Met Arg Gln Arg Gln Leu Phe             500 505 510 Glu Glu Gln Ala Lys Ala Lys Ala Ala Gly Asp Asp Glu Ala Met Phe         515 520 525 Ile Asp Glu Asn Phe Cys Thr Ala Leu Glu Tyr Gly Leu Pro Pro Thr     530 535 540 Ala Gly Trp Gly Met Gly Ile Asp Arg Val Ala Met Phe Leu Thr Asp 545 550 555 560 Ser Asn Asn Ile Lys Glu Val Leu Leu Phe Pro Ala Met Lys Pro Glu                 565 570 575 Asp Lys Lys Glu Asn Val Ala Thr Thr Asp Thr Leu Glu Ser Thr Thr             580 585 590 Val Gly Thr Ser Val         595 <210> 2 <211> 1794 <212> DNA <213> Artificial Sequence <220> <223> DNA sequence of KRS <400> 2 atggcggccg tgcaggcggc cgaggtgaaa gtggatggca gcgagccgaa actgagcaag 60 aatgagctga agagacgcct gaaagctgag aagaaagtag cagagaagga ggccaaacag 120 aaagagctca gtgagaaaca gctaagccaa gccactgctg ctgccaccaa ccacaccact 180 gataatggtg tgggtcctga ggaagagagc gtggacccaa atcaatacta caaaatccgc 240 agtcaagcaa ttcatcagct gaaggtcaat ggggaagacc catacccaca caagttccat 300 gtagacatct cactcactga cttcatccaa aaatatagtc acctgcagcc tggggatcac 360 ctgactgaca tcaccttaaa ggtggcaggt aggatccatg ccaaaagagc ttctggggga 420 aagctcatct tctatgatct tcgaggagag ggggtgaagt tgcaagtcat ggccaattcc 480 agaaattata aatcagaaga agaatttatt catattaata acaaactgcg tcggggagac 540 ataattggag ttcaggggaa tcctggtaaa accaagaagg gtgagctgag catcattccg 600 tatgagatca cactgctgtc tccctgtttg catatgttac ctcatcttca ctttgggctc 660 aaagacaagg aaacaaggta tcgccagaga tacttggact tgatcctgaa tgactttgtg 720 aggcagaaat ttatcatccg ctctaagatc atcacatata taagaagttt cttagatgag 780 ctgggattcc tagagattga aactcccatg atgaacatca tcccaggggg agccgtggcc 840 aagcctttca tcacttatca caacgagctg gacatgaact tatatatgag aattgctcca 900 gaactctatc ataagatgct tgtggttggt ggcatcgacc gggtttatga aattggacgc 960 cagttccgga atgaggggat tgatttgacg cacaatcctg agttcaccac ctgtgagttc 1020 tacatggcct atgcagacta tcacgatctc atggaaatca cggagaagat ggtttcaggg 1080 atggtgaagc atattacagg cagttacaag gtcacctacc acccagatgg cccagagggc 1140 caagcctacg atgttgactt caccccaccc ttccggcgaa tcaacatggt agaagagctt 1200 gagaaagccc tggggatgaa gctgccagaa acgaacctct ttgaaactga agaaactcgc 1260 aaaattcttg atgatatctg tgtggcaaaa gctgttgaat gccctccacc tcggaccaca 1320 gccaggctcc ttgacaagct tgttggggag ttcctggaag tgacttgcat caatcctaca 1380 ttcatctgtg atcacccaca gataatgagc cctttggcta aatggcaccg ctctaaagag 1440 ggtctgactg agcgctttga gctgtttgtc atgaagaaag agatatgcaa tgcgtatact 1500 gagctgaatg atcccatgcg gcagcggcag ctttttgaag aacaggccaa ggccaaggct 1560 gcaggtgatg atgaggccat gttcatagat gaaaacttct gtactgccct ggaatatggg 1620 ctgcccccca cagctggctg gggcatgggc attgatcgag tcgccatgtt tctcacggac 1680 tccaacaaca tcaaggaagt acttctgttt cctgccatga aacccgaaga caagaaggag 1740 aatgtagcaa ccactgatac actggaaagc acaacagttg gcacttctgt ctag 1794 <210> 3 <211> 1176 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of LRS (Homo sapiens) <400> 3 Met Ala Glu Arg Lys Gly Thr Ala Lys Val Asp Phe Leu Lys Lys Ile   1 5 10 15 Glu Lys Glu Ile Gln Gln Lys Trp Asp Thr Glu Arg Val Phe Glu Val              20 25 30 Asn Ala Ser Asn Leu Glu Lys Gln Thr Ser Lys Gly Lys Tyr Phe Val          35 40 45 Thr Phe Pro Tyr Pro Tyr Met Asn Gly Arg Leu His Leu Gly His Thr      50 55 60 Phe Ser Leu Ser Lys Cys Glu Phe Ala Val Gly Tyr Gln Arg Leu Lys  65 70 75 80 Gly Lys Cys Cys Leu Phe Pro Phe Gly Leu His Cys Thr Gly Met Pro                  85 90 95 Ile Lys Ala Cys Ala Asp Lys Leu Lys Arg Glu Ile Glu Leu Tyr Gly             100 105 110 Cys Pro Pro Asp Phe Pro Asp Glu Glu Glu Glu Glu Glu Glu Thr Ser         115 120 125 Val Lys Thr Glu Asp Ile Ile Ile Lys Asp Lys Ala Lys Gly Lys Lys     130 135 140 Ser Lys Ala Ala Ala Lys Ala Gly Ser Ser Lys Tyr Gln Trp Gly Ile 145 150 155 160 Met Lys Ser Leu Gly Leu Ser Asp Glu Glu Ile Val Lys Phe Ser Glu                 165 170 175 Ala Glu His Trp Leu Asp Tyr Phe Pro Pro Leu Ala Ile Gln Asp Leu             180 185 190 Lys Arg Met Gly Leu Lys Val Asp Trp Arg Arg Ser Phe Ile Thr Thr         195 200 205 Asp Val Asn Pro Tyr Tyr Asp Ser Phe Val Arg Trp Gln Phe Leu Thr     210 215 220 Leu Arg Glu Arg Asn Lys Ile Lys Phe Gly Lys Arg Tyr Thr Ile Tyr 225 230 235 240 Ser Pro Lys Asp Gly Gln Pro Cys Met Asp His Asp Arg Gln Thr Gly                 245 250 255 Glu Gly Val Gly Pro Gln Glu Tyr Thr Leu Leu Lys Leu Lys Val Leu             260 265 270 Glu Pro Tyr Pro Ser Lys Leu Ser Gly Leu Lys Gly Lys Asn Ile Phe         275 280 285 Leu Val Ala Ala Thr Leu Arg Pro Glu Thr Met Phe Gly Gln Thr Asn     290 295 300 Cys Trp Val Arg Pro Asp Met Lys Tyr Ile Gly Phe Glu Thr Val Asn 305 310 315 320 Gly Asp Ile Phe Ile Cys Thr Gln Lys Ala Ala Arg Asn Met Ser Tyr                 325 330 335 Gln Gly Phe Thr Lys Asp Asn Gly Val Val Pro Val Val Lys Glu Leu             340 345 350 Met Gly Glu Glu Ile Leu Gly Ala Ser Leu Ser Ala Pro Leu Thr Ser         355 360 365 Tyr Lys Val Ile Tyr Val Leu Pro Met Leu Thr Ile Lys Glu Asp Lys     370 375 380 Gly Thr Gly Val Val Thr Ser Val Pro Ser Asp Ser Pro Asp Asp Ile 385 390 395 400 Ala Ala Leu Arg Asp Leu Lys Lys Lys Lys Gln Ala Leu Arg Ala Lys Tyr                 405 410 415 Gly Ile Arg Asp Asp Met Val Leu Pro Phe Glu Pro Val Pro Val Ile             420 425 430 Glu Ile Pro Gly Phe Gly Asn Leu Ser Ala Val Thr Ile Cys Asp Glu         435 440 445 Leu Lys Ile Gln Ser Gln Asn Asp Arg Glu Lys Leu Ala Glu Ala Lys     450 455 460 Glu Lys Ile Tyr Leu Lys Gly Phe Tyr Glu Gly Ile Met Leu Val Asp 465 470 475 480 Gly Phe Lys Gly Gln Lys Val Gln Asp Val Lys Lys Thr Ile Gln Lys                 485 490 495 Lys Met Ile Asp Ala Gly Asp Ala Leu Ile Tyr Met Glu Pro Glu Lys             500 505 510 Gln Val Met Ser Arg Ser Ser Asp Glu Cys Val Val Ala Leu Cys Asp         515 520 525 Gln Trp Tyr Leu Asp Tyr Gly Glu Glu Asn Trp Lys Lys Gln Thr Ser     530 535 540 Gln Cys Leu Lys Asn Leu Glu Thr Phe Cys Glu Glu Thr Arg Arg Asn 545 550 555 560 Phe Glu Ala Thr Leu Gly Trp Leu Gln Glu His Ala Cys Ser Arg Thr                 565 570 575 Tyr Gly Leu Gly Thr His Leu Pro Trp Asp Glu Gln Trp Leu Ile Glu             580 585 590 Ser Leu Ser Asp Ser Thr Ile Tyr Met Ala Phe Tyr Thr Val Ala His         595 600 605 Leu Leu Gln Gly Gly Asn Leu His Gly Gln Ala Glu Ser Pro Leu Gly     610 615 620 Ile Arg Pro Gln Gln Met Thr Lys Glu Val Trp Asp Tyr Val Phe Phe 625 630 635 640 Lys Glu Ala Pro Phe Pro Lys Thr Gln Ile Ala Lys Glu Lys Leu Asp                 645 650 655 Gln Leu Lys Gln Glu Phe Glu Phe Trp Tyr Pro Val Asp Leu Arg Val             660 665 670 Ser Gly Lys Asp Leu Val Pro Asn His Leu Ser Tyr Tyr Leu Tyr Asn         675 680 685 His Val Ala Met Trp Pro Glu Gln Ser Asp Lys Trp Pro Thr Ala Val     690 695 700 Arg Ala Asn Gly His Leu Leu Leu Asn Ser Glu Lys Met Ser Lys Ser 705 710 715 720 Thr Gly Asn Phe Leu Thr Leu Thr Gln Ala Ile Asp Lys Phe Ser Ala                 725 730 735 Asp Gly Met Arg Leu Ala Leu Ala Asp Ala Gly Asp Thr Val Glu Asp             740 745 750 Ala Asn Phe Val Glu Ala Met Ala Asp Ala Gly Ile Leu Arg Leu Tyr         755 760 765 Thr Trp Val Glu Trp Val Lys Glu Met Val Ala Asn Trp Asp Ser Leu     770 775 780 Arg Ser Gly Pro Ala Ser Thr Phe Asn Asp Arg Val Phe Ala Ser Glu 785 790 795 800 Leu Asn Ala Gly Ile Ile Lys Thr Asp Gln Asn Tyr Glu Lys Met Met                 805 810 815 Phe Lys Glu Ala Leu Lys Thr Gly Phe Phe Glu Phe Gln Ala Ala Lys             820 825 830 Asp Lys Tyr Arg Glu Leu Ala Val Glu Gly Met His Arg Glu Leu Val         835 840 845 Phe Arg Phe Ile Glu Val Gln Thr Leu Leu Leu Ala Pro Phe Cys Pro     850 855 860 His Leu Cys Glu His Ile Trp Thr Leu Leu Gly Lys Pro Asp Ser Ile 865 870 875 880 Met Asn Ala Ser Trp Pro Val Ala Gly Pro Val Asn Glu Val Leu Ile                 885 890 895 His Ser Ser Gln Tyr Leu Met Glu Val Thr His Asp Leu Arg Leu Arg             900 905 910 Leu Lys Asn Tyr Met Met Pro Ala Lys Gly Lys Lys Thr Asp Lys Gln         915 920 925 Pro Leu Gln Lys Pro Ser His Cys Thr Ile Tyr Val Ala Lys Asn Tyr     930 935 940 Pro Pro Trp Gln His Thr Thr Leu Ser Val Leu Arg Lys His Phe Glu 945 950 955 960 Ala Asn Asn Gly Lys Leu Pro Asp Asn Lys Val Ile Ala Ser Glu Leu                 965 970 975 Gly Ser Met Pro Glu Leu Lys Lys Tyr Met Lys Lys Val Met Pro Phe             980 985 990 Val Ala Met Ile Lys Glu Asn Leu Glu Lys Met Gly Pro Arg Ile Leu         995 1000 1005 Asp Leu Gln Leu Glu Phe Asp Glu Lys Ala Val Leu Met Glu Asn Ile    1010 1015 1020 Val Tyr Leu Thr Asn Ser Leu Glu Leu Glu His Ile Glu Val Lys Phe 1025 1030 1035 1040 Ala Ser Glu Ala Glu Asp Lys Ile Arg Glu Asp Cys Cys Pro Gly Lys                1045 1050 1055 Pro Leu Asn Val Phe Arg Ile Glu Pro Gly Val Ser Val Ser Leu Val            1060 1065 1070 Asn Pro Gln Pro Ser Asn Gly His Phe Ser Thr Lys Ile Glu Ile Arg        1075 1080 1085 Gln Gly Asp Asn Cys Asp Ser Ile Ile Arg Arg Leu Met Lys Met Asn    1090 1095 1100 Arg Gly Ile Lys Asp Leu Ser Lys Val Lys Leu Met Arg Phe Asp Asp 1105 1110 1115 1120 Pro Leu Leu Gly Pro Arg Arg Val Pro Val Leu Gly Lys Glu Tyr Thr                1125 1130 1135 Glu Lys Thr Pro Ile Ser Glu His Ala Val Phe Asn Val Asp Leu Met            1140 1145 1150 Ser Lys Lys Ile His Leu Thr Glu Asn Gly Ile Arg Val Asp Ile Gly        1155 1160 1165 Asp Thr Ile Ile Tyr Leu Val His    1170 1175 <210> 4 <211> 1823 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit alpha-4 (Homo sapiens) <400> 4 Met Ala Leu Ser Ser Ala Trp Arg Ser Val Leu Pro Leu Trp Leu Leu   1 5 10 15 Trp Ser Ala Ala Cys Ser Arg Ala Ala Ser Gly Asp Asp Asn Ala Phe              20 25 30 Pro Phe Asp Ile Glu Gly Ser Ser Ala Val Gly Arg Gln Asp Pro Pro          35 40 45 Glu Thr Ser Glu Pro Arg Val Ala Leu Gly Arg Leu Pro Pro Ala Ala      50 55 60 Glu Lys Cys Asn Ala Gly Phe Phe His Thr Leu Ser Gly Glu Cys Val  65 70 75 80 Pro Cys Asp Cys Asn Gly Asn Ser Asn Glu Cys Leu Asp Gly Ser Gly                  85 90 95 Tyr Cys Val His Cys Gln Arg Asn Thr Thr Gly Glu His Cys Glu Lys             100 105 110 Cys Leu Asp Gly Tyr Ile Gly Asp Ser Ile Arg Gly Ala Pro Gln Phe         115 120 125 Cys Gln Pro Cys Pro Cys Pro Leu Pro His Leu Ala Asn Phe Ala Glu     130 135 140 Ser Cys Tyr Arg Lys Asn Gly Ala Val Arg Cys Ile Cys Asn Glu Asn 145 150 155 160 Tyr Ala Gly Pro Asn Cys Glu Arg Cys Ala Pro Gly Tyr Tyr Gly Asn                 165 170 175 Pro Leu Leu Ile Gly Ser Thr Cys Lys Lys Cys Asp Cys Ser Gly Asn             180 185 190 Ser Asp Pro Asn Leu Ile Phe Glu Asp Cys Asp Glu Val Thr Gly Gln         195 200 205 Cys Arg Asn Cys Leu Arg Asn Thr Thr Gly Phe Lys Cys Glu Arg Cys     210 215 220 Ala Pro Gly Tyr Tyr Gly Asp Ala Arg Ile Ala Lys Asn Cys Ala Val 225 230 235 240 Cys Asn Cys Gly Gly Gly Pro Cys Asp Ser Val Thr Gly Glu Cys Leu                 245 250 255 Glu Glu Gly Phe Glu Pro Pro Thr Gly Met Asp Cys Pro Thr Ile Ser             260 265 270 Cys Asp Lys Cys Val Trp Asp Leu Thr Asp Ala Leu Arg Leu Ala Ala         275 280 285 Leu Ser Ile Glu Glu Gly Lys Ser Gly Val Leu Ser Val Ser Ser Gly     290 295 300 Ala Ala Ala His Arg His Val Asn Glu Ile Asn Ala Thr Ile Tyr Leu 305 310 315 320 Leu Lys Thr Lys Leu Ser Glu Arg Glu Asn Gln Tyr Ala Leu Arg Lys                 325 330 335 Ile Gln Ile Asn Asn Ala Glu Asn Thr Met Lys Ser Leu Leu Ser Asp             340 345 350 Val Glu Glu Leu Val Glu Lys Glu Asn Gln Ala Ser Arg Lys Gly Gln         355 360 365 Leu Val Gln Lys Glu Ser Met Asp Thr Ile Asn His Ala Ser Gln Leu     370 375 380 Val Glu Gln Ala His Asp Met Arg Asp Lys Ile Gln Glu Ile Asn Asn 385 390 395 400 Lys Met Leu Tyr Tyr Gly Glu Glu His Glu Leu Ser Pro Lys Glu Ile                 405 410 415 Ser Glu Lys Leu Val Leu Ala Gln Lys Met Leu Glu Glu Ile Arg Ser             420 425 430 Arg Gln Pro Phe Phe Thr Gln Arg Glu Leu Val Asp Glu Glu Ala Asp         435 440 445 Glu Ala Tyr Glu Leu Leu Ser Gln Ala Glu Ser Trp Gln Arg Leu His     450 455 460 Asn Glu Thr Arg Thr Leu Phe Pro Val Val Leu Glu Gln Leu Asp Asp 465 470 475 480 Tyr Asn Ala Lys Leu Ser Asp Leu Gln Glu Ala Leu Asp Gln Ala Leu                 485 490 495 Asn Tyr Val Arg Asp Ala Glu Asp Met Asn Arg Ala Thr Ala Ala Arg             500 505 510 Gln Arg Asp His Glu Lys Gln Gln Glu Arg Val Arg Glu Gln Met Glu         515 520 525 Val Val Asn Met Ser Leu Ser Thr Ser Ala Asp Ser Leu Thr Thr Pro     530 535 540 Arg Leu Thr Leu Ser Glu Leu Asp Asp Ile Ile Lys Asn Ala Ser Gly 545 550 555 560 Ile Tyr Ala Glu Ile Asp Gly Ala Lys Ser Glu Leu Gln Val Lys Leu                 565 570 575 Ser Asn Leu Ser Asn Leu Ser His Asp Leu Val Gln Glu Ala Ile Asp             580 585 590 His Ala Gln Asp Leu Gln Gln Glu Ala Asn Glu Leu Ser Arg Lys Leu         595 600 605 His Ser Ser Asp Met Asn Gly Leu Val Gln Lys Ala Leu Asp Ala Ser     610 615 620 Asn Val Tyr Glu Asn Ile Val Asn Tyr Val Ser Glu Ala Asn Glu Thr 625 630 635 640 Ala Glu Phe Ala Leu Asn Thr Thr Asp Arg Ile Tyr Asp Ala Val Ser                 645 650 655 Gly Ile Asp Thr Gln Ile Ile Tyr His Lys Asp Glu Ser Glu Asn Leu             660 665 670 Leu Asn Gln Ala Arg Glu Leu Gln Ala Lys Ala Glu Ser Ser Ser Asp         675 680 685 Glu Ala Val Ala Asp Thr Ser Arg Arg Val Gly Gly Ala Leu Ala Arg     690 695 700 Lys Ser Ala Leu Lys Thr Arg Leu Ser Asp Ala Val Lys Gln Leu Gln 705 710 715 720 Ala Ala Glu Arg Gly Asp Ala Gln Gln Arg Leu Gly Gln Ser Arg Leu                 725 730 735 Ile Thr Glu Glu Ala Asn Arg Thr Thr Met Glu Val Gln Gln Ala Thr             740 745 750 Ala Pro Met Ala Asn Asn Leu Thr Asn Trp Ser Gln Asn Leu Gln His         755 760 765 Phe Asp Ser Ser Ala Tyr Asn Thr Ala Val Asn Ser Ala Arg Asp Ala     770 775 780 Val Arg Asn Leu Thr Glu Val Val Pro Gln Leu Leu Asp Gln Leu Arg 785 790 795 800 Thr Val Glu Gln Lys Arg Pro Ala Ser Asn Val Ser Ala Ser Ile Gln                 805 810 815 Arg Ile Arg Glu Leu Ile Ala Gln Thr Arg Ser Val Ala Ser Lys Ile             820 825 830 Gln Val Ser Met Met Phe Asp Gly Gln Ser Ala Val Glu Val His Ser         835 840 845 Arg Thr Ser Met Asp Asp Leu Lys Ala Phe Thr Ser Leu Ser Leu Tyr     850 855 860 Met Lys Pro Pro Val Lys Arg Pro Glu Leu Thr Glu Thr Ala Asp Gln 865 870 875 880 Phe Ile Leu Tyr Leu Gly Ser Lys Asn Ala Lys Lys Glu Tyr Met Gly                 885 890 895 Leu Ala Ile Lys Asn Asp Asn Leu Val Tyr Val Tyr Asn Leu Gly Thr             900 905 910 Lys Asp Val Glu Ile Pro Leu Asp Ser Lys Pro Val Ser Ser Trp Pro         915 920 925 Ala Tyr Phe Ser Ile Val Lys Ile Glu Arg Val Gly Lys His Gly Lys     930 935 940 Val Phe Leu Thr Val Pro Ser Leu Ser Ser Thr Ala Glu Glu Lys Phe 945 950 955 960 Ile Lys Lys Gly Glu Phe Ser Gly Asp Asp Ser Leu Leu Asp Leu Asp                 965 970 975 Pro Glu Asp Thr Val Phe Tyr Val Gly Gly Val Pro Ser Asn Phe Lys             980 985 990 Leu Pro Thr Ser Leu Asn Leu Pro Gly Phe Val Gly Cys Leu Glu Leu         995 1000 1005 Ala Thr Leu Asn Asn Asp Val Ile Ser Leu Tyr Asn Phe Lys His Ile    1010 1015 1020 Tyr Asn Met Asp Pro Ser Thr Ser Val Pro Cys Ala Arg Asp Lys Leu 1025 1030 1035 1040 Ala Phe Thr Gln Ser Arg Ala Ala Ser Tyr Phe Phe Asp Gly Ser Gly                1045 1050 1055 Tyr Ala Val Val Arg Asp Ile Thr Arg Arg Gly Lys Phe Gly Gln Val            1060 1065 1070 Thr Arg Phe Asp Ile Glu Val Arg Thr Pro Ala Asp Asn Gly Leu Ile        1075 1080 1085 Leu Leu Met Val Asn Gly Ser Met Phe Phe Arg Leu Glu Met Arg Asn    1090 1095 1100 Gly Tyr Leu His Val Phe Tyr Asp Phe Gly Phe Ser Gly Gly Pro Val 1105 1110 1115 1120 His Leu Glu Asp Thr Leu Lys Lys Ala Gln Ile Asn Asp Ala Lys Tyr                1125 1130 1135 His Glu Ile Ser Ile Ile Tyr His Asn Asp Lys Lys Met Ile Leu Val            1140 1145 1150 Val Asp Arg Arg His Val Lys Ser Met Asp Asn Glu Lys Met Lys Ile        1155 1160 1165 Pro Phe Thr Asp Ile Tyr Ile Gly Aly Pro Pro Glu Ile Leu Gln    1170 1175 1180 Ser Arg Ala Leu Arg Ala His Leu Pro Leu Asp Ile Asn Phe Arg Gly 1185 1190 1195 1200 Cys Met Lys Gly Phe Gln Phe Gln Lys Lys Asp Phe Asn Leu Leu Glu                1205 1210 1215 Gln Thr Glu Thr Leu Gly Val Gly Tyr Gly Cys Pro Glu Asp Ser Leu            1220 1225 1230 Ile Ser Arg Arg Ala Tyr Phe Asn Gly Gln Ser Phe Ile Ala Ser Ile        1235 1240 1245 Gln Lys Ile Ser Phe Phe Asp Gly Phe Glu Gly Gly Phe Asn Phe Arg    1250 1255 1260 Thr Leu Gln Pro Asn Gly Leu Leu Phe Tyr Tyr Ala Ser Gly Ser Asp 1265 1270 1275 1280 Val Phe Ser Ile Ser Leu Asp Asn Gly Thr Val Ile Met Asp Val Lys                1285 1290 1295 Gly Ile Lys Val Gln Ser Val Asp Lys Gln Tyr Asn Asp Gly Leu Ser            1300 1305 1310 His Phe Val Ile Ser Ser Val Ser Pro Thr Arg Tyr Glu Leu Ile Val        1315 1320 1325 Asp Lys Ser Arg Val Gly Ser Lys Asn Pro Thr Lys Gly Lys Ile Glu    1330 1335 1340 Gln Thr Gln Ala Ser Glu Lys Lys Phe Tyr Phe Gly Gly Ser Pro Ile 1345 1350 1355 1360 Ser Ala Gln Tyr Ala Asn Phe Thr Gly Cys Ile Ser Asn Ala Tyr Phe                1365 1370 1375 Thr Arg Val Asp Arg Asp Val Glu Val Glu Asp Phe Gln Arg Tyr Thr            1380 1385 1390 Glu Lys Val His Thr Ser Leu Tyr Glu Cys Pro Ile Glu Ser Ser Pro        1395 1400 1405 Leu Phe Leu Leu His Lys Lys Gly Lys Asn Leu Ser Lys Pro Lys Ala    1410 1415 1420 Ser Gln Asn Lys Lys Gly Gly Lys Ser Lys Asp Ala Pro Ser Trp Asp 1425 1430 1435 1440 Pro Val Ala Leu Lys Leu Pro Glu Arg Asn Thr Pro Arg Asn Ser His                1445 1450 1455 Cys His Leu Ser Asn Ser Pro Arg Ala Ile Glu His Ala Tyr Gln Tyr            1460 1465 1470 Gly Gly Thr Ala Asn Ser Arg Gln Glu Phe Glu His Leu Lys Gly Asp        1475 1480 1485 Phe Gly Ala Lys Ser Gln Phe Ser Ile Arg Leu Arg Thr Arg Ser Ser    1490 1495 1500 His Gly Met Ile Phe Tyr Val Ser Asp Gln Glu Glu Asn Asp Phe Met 1505 1510 1515 1520 Thr Leu Phe Leu Ala His Gly Arg Leu Val Tyr Met Phe Asn Val Gly                1525 1530 1535 His Lys Lys Leu Lys Ile Arg Ser Gln Glu Lys Tyr Asn Asp Gly Leu            1540 1545 1550 Trp His Asp Val Ile Phe Ile Arg Glu Arg Ser Ser Gly Arg Leu Val        1555 1560 1565 Ile Asp Gly Leu Arg Val Leu Glu Glu Ser Leu Pro Pro Thr Glu Ala    1570 1575 1580 Thr Trp Lys Ile Lys Gly Pro Ile Tyr Leu Gly Gly Val Ala Pro Gly 1585 1590 1595 1600 Lys Ala Val Lys Asn Val Gln Ile Asn Ser Ile Tyr Ser Phe Ser Gly                1605 1610 1615 Cys Leu Ser Asn Leu Gln Leu Asn Gly Ala Ser Ile Thr Ser Ala Ser            1620 1625 1630 Gln Thr Phe Ser Val Thr Pro Cys Phe Glu Gly Pro Met Glu Thr Gly        1635 1640 1645 Thr Tyr Phe Ser Thr Glu Gly Gly Tyr Val Val Leu Asp Glu Ser Phe    1650 1655 1660 Asn Ile Gly Leu Lys Phe Glu Ile Ala Phe Glu Val Arg Pro Arg Ser 1665 1670 1675 1680 Ser Ser Gly Thr Leu Val His Gly His Ser Val Asn Gly Glu Tyr Leu                1685 1690 1695 Asn Val His Met Lys Asn Gly Gln Val Ile Val Lys Val Asn Asn Gly            1700 1705 1710 Ile Arg Asp Phe Ser Thr Ser Val Thr Pro Lys Gln Ser Leu Cys Asp        1715 1720 1725 Gly Arg Trp His Arg Ile Thr Val Ile Arg Asp Ser Asn Val Val Gln    1730 1735 1740 Leu Asp Val Asp Ser Glu Val Asn His Val Val Gly Pro Leu Asn Pro 1745 1750 1755 1760 Lys Pro Ile Asp His Arg Glu Pro Val Phe Val Gly Gly Val Pro Glu                1765 1770 1775 Ser Leu Leu Thr Pro Arg Leu Ala Pro Ser Lys Pro Phe Thr Gly Cys            1780 1785 1790 Ile Arg His Phe Val Ile Asp Gly His Pro Val Ser Phe Ser Lys Ala        1795 1800 1805 Ala Leu Val Ser Gly Ala Val Ser Ile Asn Ser Cys Pro Ala Ala    1810 1815 1820 <210> 5 <211> 5472 <212> DNA <213> Artificial Sequence <220> <223> DNA sequence of Laminin subunit alpha-4 (Homo sapiens) <400> 5 atggctttga gctcagcctg gcgctcggtt ctgcctctgt ggctcctctg gagcgctgcc 60 tgctcccgcg ccgcgtccgg ggacgacaac gcttttcctt ttgacattga agggagctca 120 gcggttggca ggcaagaccc gcctgagacg agcgaacccc gcgtggctct gggacgcctg 180 ccgcctgcgg ccgagaaatg caatgctgga ttctttcaca ccctgtcggg agaatgtgtg 240 ccctgcgact gtaatggcaa ttccaacgag tgtttggacg gctcaggata ctgtgtgcac 300 tgccagcgga acacaacagg agagcactgt gaaaagtgtc tggatggtta tatcggagat 360 tccatcaggg gagcacccca attctgccag ccgtgcccct gtcccctgcc ccacttggcc 420 aattttgcag aatcctgcta taggaaaaat ggagctgttc ggtgcatttg taacgaaaat 480 tatgctggac ctaactgtga aagatgtgct cccggttact atggaaaccc cttactcatt 540 ggaagcacct gtaagaaatg tgactgcagt ggaaattcag atcccaacct gatctttgaa 600 gattgtgatg aagtcactgg ccagtgtagg aattgcttac gcaacaccac cggattcaag 660 tgtgaacgtt gcgctcctgg ctactatggg gacgccagga tagccaagaa ctgtgcagtg 720 tgcaactgcg ggggaggccc atgtgacagt gtaaccggag aatgcttgga agaaggtttt 780 gaacccccta caggcatgga ctgcccaacc ataagctgtg ataagtgcgt ctgggacctg 840 actgatgacc tgcggttagc agcgctctcc atcgaggaag gcaaatccgg ggtgctgagc 900 gtatcctctg gggccgccgc tcataggcac gtgaatgaaa tcaacgccac catctacctc 960 ctcaaaacaa aattgtcaga aagagaaaac caatacgccc taagaaagat acaaatcaac 1020 aatgctgaga acacgatgaa aagccttctg tctgacgtag aggaattagt tgaaaaggaa 1080 aatcaagcct ccagaaaagg acaacttgtt cagaaggaaa gcatggacac cattaaccac 1140 gcaagtcagc tggtagagca agcccatgat atgagggata aaatccaaga gatcaacaac 1200 aagatgctct attatgggga agagcatgaa cttagcccca aggaaatctc tgagaagctg 1260 gtgttggccc agaagatgct tgaagagatt agaagccgtc aaccattttt cacccaacgg 1320 gagctcgtgg atgaggaggc agatgaggct tacgaactac tgagccaggc tgagagctgg 1380 cagcggctgc acaatgagac ccgcactctg tttcctgtcg tcctggagca gctggatgac 1440 tacaatgcta agttgtcaga tctccaggaa gcacttgacc aggcccttaa ctatgtcagg 1500 gatgccgaag acatgaacag ggccacagca gccaggcagc gggaccatga gaaacaacag 1560 gaaagagtga gggaacaaat ggaagtggtg aacatgtctc tgagcacatc tgcggactct 1620 ctgacaacac ctcgtctaac tctttcagaa cttgatgata taataaagaa tgcgtcaggg 1680 atttatgcag aaatagatgg agccaaaagt gaactacaag taaaactatc taacctaagt 1740 aacctcagcc atgatttagt ccaagaagct attgaccatg cacaggacct tcaacaagaa 1800 gctaatgaat tgagcaggaa gttgcacagt tcagatatga acgggctggt acagaaggct 1860 ttggatgcat caaatgtcta tgaaaatatt gttaattatg ttagtgaagc caatgaaaca 1920 gcagaatttg ctttgaacac cactgaccga atttatgatg cggtgagtgg gattgatact 1980 caaatcattt accataaaga tgaaagtgag aacctcctca atcaagccag agaactgcaa 2040 gcaaaggcag agtctagcag tgatgaagca gtggctgaca ctagcaggcg tgtgggtgga 2100 gccctagcaa ggaaaagtgc ccttaaaacc agactcagtg atgccgttaa gcaactacaa 2160 gcagcagaga gaggggatgc ccagcagcgc ctggggcagt ctagactgat caccgaggaa 2220 gccaacagga cgacgatgga ggtgcagcag gccactgccc ccatggccaa caatctaacc 2280 aactggtcac agaatcttca acattttgac tcttctgctt acaacactgc agtgaactct 2340 gctagggatg cagtaagaaa tctgaccgag gttgtccctc agctcctgga tcagcttcgt 2400 acggttgagc agaagcgacc tgcaagcaac gtttctgcca gcatccagag gatccgagag 2460 ctcattgctc agaccagaag tgttgccagc aagatccaag tctccatgat gtttgatggc 2520 cagtcagctg tggaagtgca ctcgagaacc agtatggatg acttaaaggc cttcacgtct 2580 ctgagcctgt acatgaaacc ccctgtgaag cggccggaac tgaccgagac tgcagatcag 2640 tttatcctgt acctcggaag caaaaacgcc aaaaaagagt atatgggtct tgcaatcaaa 2700 aatgataatc tggtatacgt ctataatttg ggaactaaag atgtggagat tcccctggac 2760 tccaagcccg tcagttcctg gcctgcttac ttcagcattg tcaagattga aagggtggga 2820 aaacatggaa aggtgttttt aacagtcccg agtctaagta gcacagcaga ggaaaagttc 2880 attaaaaagg gggaattttc gggagatgac tctctgctgg acctggaccc tgaggacaca 2940 gtgttttatg ttggtggagt gccttccaac ttcaagctcc ctaccagctt aaacctgcct 3000 ggctttgttg gctgcctgga actggccact ttgaataatg atgtgatcag cttgtacaac 3060 tttaagcaca tctataatat ggacccctcc acatcagtgc catgtgcccg agataagctg 3120 gccttcactc agagtcgggc tgccagttac ttcttcgatg gctccggtta tgccgtggtg 3180 agagacatca caaggagagg gaaatttggt caggtgactc gctttgacat agaagttcga 3240 acaccagctg acaacggcct tattctcctg atggtcaatg gaagtatgtt tttcagactg 3300 gaaatgcgca atggttacct acatgtgttc tatgattttg gattcagcgg tggccctgtg 3360 catcttgaag atacgttaaa gaaagctcaa attaatgatg caaaatacca tgagatctca 3420 atcatttacc acaatgataa gaaaatgatc ttggtagttg acagaaggca tgtcaagagc 3480 atggataatg aaaagatgaa aatacctttt acagatatat acattggagg agctcctcca 3540 gaaatcttac aatccagggc cctcagagca caccttcccc tagatatcaa cttcagagga 3600 tgcatgaagg gcttccagtt ccaaaagaag gacttcaatt tactggagca gacagaaacc 3660 ctgggagttg gttatggatg cccagaagac tcacttatat ctcgcagagc atatttcaat 3720 ggacagagct tcattgcttc aattcagaaa atatctttct ttgatggctt tgaaggaggt 3780 tttaatttcc gaacattaca accaaatggg ttactattct attatgcttc agggtcagac 3840 gtgttctcca tctcactgga taatggtact gtcatcatgg atgtaaaggg aatcaaagtt 3900 cagtcagtag ataagcagta caatgatggg ctgtcccact tcgtcattag ctctgtctca 3960 cccacaagat atgaactgat agtagataaa agcagagttg ggagtaagaa tcctaccaaa 4020 gggaaaatag aacagacaca agcaagtgaa aagaagtttt acttcggtgg ctcaccaatc 4080 agtgctcagt atgctaattt cactggctgc ataagtaatg cctactttac cagggtggat 4140 agagatgtgg aggttgaaga tttccaacgg tatactgaaa aggtccacac ttctctttat 4200 gagtgtccca ttgagtcttc accattgttt ctcctccata aaaaaggaaa aaatttatcc 4260 aagcctaaag caagtcagaa taaaaaggga gggaaaagta aagatgcacc ttcatgggat 4320 cctgttgctc tgaaactccc agagcggaat actccaagaa actctcattg ccacctttcc 4380 aacagcccta gagcaataga gcacgcctat caatatggag gaacagccaa cagccgccaa 4440 gagtttgaac acttaaaagg agattttggt gccaaatctc agttttccat tcgtctgaga 4500 actcgttcct cccatggcat gatcttctat gtctcagatc aagaagagaa tgacttcatg 4560 actctatttt tggcccatgg ccgcttggtt tacatgttta atgttggtca caaaaaactg 4620 aagattagaa gccaggagaa atacaatgat ggcctgtggc atgatgtgat atttattcga 4680 gaaaggagca gtggccgact ggtaattgat ggtctccgag tcctagaaga aagtcttcct 4740 cctactgaag ctacctggaa aatcaagggt cccatttatt tgggaggtgt ggctcctgga 4800 aaggctgtga aaaatgttca gattaactcc atctacagtt ttagtggctg tctcagcaat 4860 ctccagctca atggggcctc catcacctct gcttctcaga cattcagtgt gaccccttgc 4920 tttgaaggcc ccatggaaac aggaacttac ttttcaacag aaggaggata cgtggttcta 4980 gatgaatctt tcaatattgg attgaagttt gaaattgcat ttgaagtccg tcccagaagc 5040 agttccggaa ccctggtcca cggccacagt gtcaatgggg agtacctaaa tgttcacatg 5100 aaaaatggac aggtcatagt gaaagtcaat aatggcatca gagatttttc cacctcagtt 5160 acacccaagc agagtctctg tgatggcaga tggcacagaa ttacagttat tagagattct 5220 aatgtggttc agttggatgt ggactctgaa gtgaaccatg tggttggacc cctgaatcca 5280 aaaccaattg atcacaggga gcctgtgttt gttggaggtg ttccagaatc tctactgaca 5340 ccacgcttgg cccccagcaa acccttcaca ggctgcatac gccactttgt gattgatgga 5400 cacccagtga gcttcagtaa agcagccctg gtcagcggcg ccgtaagcat caactcctgt 5460 ccagcagcct ga 5472 <210> 6 <211> 1798 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit beta-2 (Homo sapiens) <400> 6 Met Glu Leu Thr Ser Arg Glu Arg Gly Arg Gly Gln Pro Leu Pro Trp   1 5 10 15 Glu Leu Arg Leu Gly Leu Leu Leu Ser Val Leu Ala Ala Thr Leu Ala              20 25 30 Gln Ala Pro Ala Pro Asp Val Pro Gly Cys Ser Arg Gly Ser Cys Tyr          35 40 45 Pro Ala Thr Gly Asp Leu Leu Val Gly Arg Ala Asp Arg Leu Thr Ala      50 55 60 Ser Ser Thr Cys Gly Leu Asn Gly Pro Gln Pro Tyr Cys Ile Val Ser  65 70 75 80 His Leu Gln Asp Glu Lys Lys Cys Phe Leu Cys Asp Ser Arg Arg Pro                  85 90 95 Phe Ser Ala Arg Asp Asn Pro His Ser His Arg Ile Gln Asn Val Val             100 105 110 Thr Ser Phe Ala Pro Gln Arg Arg Ala Ala Trp Trp Gln Ser Glu Asn         115 120 125 Gly Ile Pro Ala Val Thr Ile Gln Leu Asp Leu Glu Ala Glu Phe His     130 135 140 Phe Thr His Leu Ile Met Thr Phe Lys Thr Phe Arg Pro Ala Ala Met 145 150 155 160 Leu Val Glu Arg Ser Ala Asp Phe Gly Arg Thr Trp His Val Tyr Arg                 165 170 175 Tyr Phe Ser Tyr Asp Cys Gly Ala Asp Phe Pro Gly Val Pro Leu Ala             180 185 190 Pro Pro Arg His Trp Asp Asp Val Val Cys Glu Ser Arg Tyr Ser Glu         195 200 205 Ile Glu Pro Ser Thr Glu Gly Glu Val Ile Tyr Arg Val Leu Asp Pro     210 215 220 Ala Ile Pro Ile Pro Asp Pro Tyr Ser Ser Arg Ile Gln Asn Leu Leu 225 230 235 240 Lys Ile Thr Asn Leu Arg Val Asn Leu Thr Arg Leu His Thr Leu Gly                 245 250 255 Asp Asn Leu Leu Asp Pro Arg Arg Glu Ile Arg Glu Lys Tyr Tyr Tyr             260 265 270 Ala Leu Tyr Glu Leu Val Val Arg Gly Asn Cys Phe Cys Tyr Gly His         275 280 285 Ala Ser Glu Cys Ala Pro Ala Pro Gly Ala Pro Ala His Ala Glu Gly     290 295 300 Met Val His Gly Ala Cys Ile Cys Lys His Asn Thr Arg Gly Leu Asn 305 310 315 320 Cys Glu Gln Cys Gln Asp Phe Tyr Arg Asp Leu Pro Trp Arg Pro Ala                 325 330 335 Glu Asp Gly His Ser His Ala Cys Arg Lys Cys Glu Cys His Gly His             340 345 350 Thr His Ser Cys His Phe Asp Met Ala Val Tyr Leu Ala Ser Gly Asn         355 360 365 Val Ser Gly Gly Val Cys Asp Gly Cys Gln His Asn Thr Ala Gly Arg     370 375 380 His Cys Glu Leu Cys Arg Pro Phe Phe Tyr Arg Asp Pro Thr Lys Asp 385 390 395 400 Leu Arg Asp Pro Ala Val Cys Arg Ser Cys Asp Cys Asp Pro Met Gly                 405 410 415 Ser Gln Asp Gly Gly Arg Cys Asp Ser His Asp Asp Pro Ala Leu Gly             420 425 430 Leu Val Ser Gly Gln Cys Arg Cys Lys Glu His Val Val Gly Thr Arg         435 440 445 Cys Gln Gln Cys Arg Asp Gly Phe Phe Gly Leu Ser Ile Ser Asp Arg     450 455 460 Leu Gly Cys Arg Arg Cys Gln Cys Asn Ala Arg Gly Thr Val Pro Gly 465 470 475 480 Ser Thr Pro Cys Asp Pro Asn Ser Gly Ser Cys Tyr Cys Lys Arg Leu                 485 490 495 Val Thr Gly Arg Gly Cys Asp Arg Cys Leu Pro Gly His Trp Gly Leu             500 505 510 Ser His Asp Leu Leu Gly Cys Arg Pro Cys Asp Cys Asp Val Gly Gly         515 520 525 Ala Leu Asp Pro Gln Cys Asp Glu Gly Thr Gly Gln Cys His Cys Arg     530 535 540 Gln His Met Val Gly Arg Arg Cys Glu Gln Val Gln Pro Gly Tyr Phe 545 550 555 560 Arg Pro Phe Leu Asp His Leu Ile Trp Glu Ala Glu Asp Thr Arg Gly                 565 570 575 Gln Val Leu Asp Val Val Glu Arg Leu Val Thr Pro Gly Glu Thr Pro             580 585 590 Ser Trp Thr Gly Ser Gly Phe Val Arg Leu Gln Glu Gly Gln Thr Leu         595 600 605 Glu Phe Leu Val Ala Ser Val Pro Lys Ala Met Asp Tyr Asp Leu Leu     610 615 620 Leu Arg Leu Glu Pro Gln Val Pro Glu Gln Trp Ala Glu Leu Glu Leu 625 630 635 640 Ile Val Gln Arg Pro Gly Pro Val Pro Ala His Ser Leu Cys Gly His                 645 650 655 Leu Val Pro Lys Asp Asp Arg Ile Gln Gly Thr Leu Gln Pro His Ala             660 665 670 Arg Tyr Leu Ile Phe Pro Asn Pro Val Cys Leu Glu Pro Gly Ile Ser         675 680 685 Tyr Lys Leu His Leu Lys Leu Val Arg Thr Gly Gly Ser Ala Gln Pro     690 695 700 Glu Thr Pro Tyr Ser Gly Pro Gly Leu Leu Ile Asp Ser Leu Val Leu 705 710 715 720 Leu Pro Arg Val Leu Val Leu Glu Met Phe Ser Gly Gly Asp Ala Ala                 725 730 735 Ala Leu Glu Arg Gln Ala Thr Phe Glu Arg Tyr Gln Cys His Glu Glu             740 745 750 Gly Leu Val Pro Ser Lys Thr Ser Pro Ser Glu Ala Cys Ala Pro Leu         755 760 765 Leu Ile Ser Leu Ser Thr Leu Ile Tyr Asn Gly Ala Leu Pro Cys Gln     770 775 780 Cys Asn Pro Gln Gly Ser Leu Ser Ser Glu Cys Asn Pro His Gly Gly 785 790 795 800 Gln Cys Leu Cys Lys Pro Gly Val Val Gly Arg Arg Cys Asp Leu Cys                 805 810 815 Ala Pro Gly Tyr Tyr Gly Phe Gly Pro Thr Gly Cys Gln Ala Cys Gln             820 825 830 Cys Ser His Glu Gly Ala Leu Ser Ser Leu Cys Glu Lys Thr Ser Gly         835 840 845 Gln Cys Leu Cys Arg Thr Gly Ala Phe Gly Leu Arg Cys Asp Arg Cys     850 855 860 Gln Arg Gly Gln Trp Gly Phe Pro Ser Cys Arg Pro Cys Val Cys Asn 865 870 875 880 Gly His Ala Asp Glu Cys Asn Thr His Thr Gly Ala Cys Leu Gly Cys                 885 890 895 Arg Asp His Thr Gly Gly Glu His Cys Glu Arg Cys Ile Ala Gly Phe             900 905 910 His Gly Asp Pro Arg Leu Pro Tyr Gly Gly Gln Cys Arg Pro Cys Pro         915 920 925 Cys Pro Glu Gly Pro Gly Ser Gln Arg His Phe Ala Thr Ser Cys His     930 935 940 Gln Asp Glu Tyr Ser Gln Gln Ile Val Cys His Cys Arg Ala Gly Tyr 945 950 955 960 Thr Gly Leu Arg Cys Glu Ala Cys Ala Pro Gly His Phe Gly Asp Pro                 965 970 975 Ser Arg Pro Gly Gly Arg Cys Gln Leu Cys Glu Cys Ser Gly Asn Ile             980 985 990 Asp Pro Met Asp Pro Asp Ala Cys Asp Pro His Thr Gly Gln Cys Leu         995 1000 1005 Arg Cys Leu His His Thr Glu Gly Pro His Cys Ala His Cys Lys Pro    1010 1015 1020 Gly Phe His Gly Gln Ala Ala Arg Gln Ser Cys His Arg Cys Thr Cys 1025 1030 1035 1040 Asn Leu Leu Gly Thr Asn Pro Gln Gln Cys Pro Ser Pro Asp Gln Cys                1045 1050 1055 His Cys Asp Pro Ser Ser Gly Gln Cys Pro Cys Leu Pro Asn Val Gln            1060 1065 1070 Gly Pro Ser Cys Asp Arg Cys Ala Pro Asn Phe Trp Asn Leu Thr Ser        1075 1080 1085 Gly His Gly Cys Gln Pro Cys Ala Cys His Pro Ser Arg Ala Arg Gly    1090 1095 1100 Pro Thr Cys Asn Glu Phe Thr Gly Gln Cys His Cys Arg Ala Gly Phe 1105 1110 1115 1120 Gly Gly Arg Thr Cys Ser Glu Cys Gln Glu Leu His Trp Gly Asp Pro                1125 1130 1135 Gly Leu Gln Cys His Ala Cys Asp Cys Asp Ser Arg Gly Ile Asp Thr            1140 1145 1150 Pro Gln Cys His Arg Phe Thr Gly His Cys Ser Cys Arg Pro Gly Val        1155 1160 1165 Ser Gly Val Arg Cys Asp Gln Cys Ala Arg Gly Phe Ser Gly Ile Phe    1170 1175 1180 Pro Ala Cys His Pro Cys His Ala Cys Phe Gly Asp Trp Asp Arg Val 1185 1190 1195 1200 Val Gln Asp Leu Ala Ala Arg Thr Gln Arg Leu Glu Gln Arg Ala Gln                1205 1210 1215 Glu Leu Gln Gln Thr Gly Val Leu Gly Ala Phe Glu Ser Ser Phe Trp            1220 1225 1230 His Met Gln Glu Lys Leu Gly Ile Val Gln Gly Ile Val Gly Ala Arg        1235 1240 1245 Asn Thr Ser Ala Ala Ser Thr Ala Gln Leu Val Glu Ala Thr Glu Glu    1250 1255 1260 Leu Arg Arg Glu Ile Gly Glu Ala Thr Glu His Leu Thr Gln Leu Glu 1265 1270 1275 1280 Ala Asp Leu Thr Asp Val Gln Asp Glu Asn Phe Asn Ala Asn His Ala                1285 1290 1295 Leu Ser Gly Leu Glu Arg Asp Arg Leu Ala Leu Asn Leu Thr Leu Arg            1300 1305 1310 Gln Leu Asp Gln His Leu Asp Leu Leu Lys His Ser Asn Phe Leu Gly        1315 1320 1325 Ala Tyr Asp Ser Ile Arg His Ala His Ser Gln Ser Ala Glu Ala Glu    1330 1335 1340 Arg Arg Ala Asn Thr Ser Ala Leu Ala Val Pro Ser Pro Val Ser Asn 1345 1350 1355 1360 Ser Ala Ser Ala Arg His Arg Thr Glu Ala Leu Met Asp Ala Gln Lys                1365 1370 1375 Glu Asp Phe Asn Ser Lys His Met Ala Asn Gln Arg Ala Leu Gly Lys            1380 1385 1390 Leu Ser Ala His Thr His Thr Leu Ser Leu Thr Asp Ile Asn Glu Leu        1395 1400 1405 Val Cys Gly Ala Pro Gly Asp Ala Pro Cys Ala Thr Ser Pro Cys Gly    1410 1415 1420 Gly Ala Gly Cys Arg Asp Glu Asp Gly Gln Pro Arg Cys Gly Gly Leu 1425 1430 1435 1440 Ser Cys Asn Gly Ala Ala Ala Thr Ala Asp Leu Ala Leu Gly Arg Ala                1445 1450 1455 Arg His Thr Gln Ala Glu Leu Gln Arg Ala Leu Ala Glu Gly Gly Ser            1460 1465 1470 Ile Leu Ser Arg Val Ala Glu Thr Arg Arg Gln Ala Ser Glu Ala Gln        1475 1480 1485 Gln Arg Ala Gln Ala Ala Leu Asp Lys Ala Asn Ala Ser Arg Gly Gln    1490 1495 1500 Val Glu Gln Ala Asn Gln Glu Leu Gln Glu Leu Ile Gln Ser Val Lys 1505 1510 1515 1520 Asp Phe Leu Asn Gln Glu Gly Ala Asp Pro Asp Ser Ile Glu Met Val                1525 1530 1535 Ala Thr Arg Val Leu Glu Leu Ser Ile Pro Ala Ser Ala Glu Gln Ile            1540 1545 1550 Gln His Leu Ala Gly Ala Ile Ala Glu Arg Val Arg Ser Leu Ala Asp        1555 1560 1565 Val Asp Ala Ile Leu Ala Arg Thr Val Gly Asp Val Arg Arg Ala Glu    1570 1575 1580 Gln Leu Leu Gln Asp Ala Arg Arg Ala Arg Ser Trp Ala Glu Asp Glu 1585 1590 1595 1600 Lys Gln Lys Ala Glu Thr Val Gln Ala Ala Leu Glu Glu Ala Gln Arg                1605 1610 1615 Ala Gln Gly Ile Ala Gln Gly Ala Ile Arg Gly Ala Val Ala Asp Thr            1620 1625 1630 Arg Asp Thr Glu Gln Thr Leu Tyr Gln Val Gln Glu Arg Met Ala Gly        1635 1640 1645 Ala Glu Arg Ala Leu Ser Ser Ala Gly Glu Arg Ala Arg Gln Leu Asp    1650 1655 1660 Ala Leu Leu Glu Ala Leu Lys Leu Lys Arg Ala Gly Asn Ser Leu Ala 1665 1670 1675 1680 Ala Ser Thr Ala Glu Glu Thr Ala Gly Ser Ala Gln Gly Arg Ala Gln                1685 1690 1695 Glu Ala Glu Gln Leu Leu Arg Gly Pro Leu Gly Asp Gln Tyr Gln Thr            1700 1705 1710 Val Lys Ala Leu Ala Glu Arg Lys Ala Gln Gly Val Leu Ala Ala Gln        1715 1720 1725 Ala Arg Ala Glu Gln Leu Arg Asp Glu Ala Arg Asp Leu Leu Gln Ala    1730 1735 1740 Ala Gln Asp Lys Leu Gln Arg Leu Gln Glu Leu Glu Gly Thr Tyr Glu 1745 1750 1755 1760 Glu Asn Glu Arg Ala Leu Glu Ser Lys Ala Ala Gln Leu Asp Gly Leu                1765 1770 1775 Glu Ala Arg Met Arg Ser Val Leu Gln Ala Ile Asn Leu Gln Val Gln            1780 1785 1790 Ile Tyr Asn Thr Cys Gln        1795 <210> 7 <211> 5397 <212> DNA <213> Artificial Sequence <220> <223> DNA sequence of Laminin subunit beta-2 (Homo sapiens) <400> 7 atggagctga cctcaaggga aagagggagg ggacagcctc tgccctggga acttcgactg 60 ggcctactgc taagcgtgct ggctgccaca ctggcacagg cccctgcccc ggatgtgcct 120 ggctgttcca ggggaagctg ctaccccgcc acgggcgacc tgctggtggg ccgagctgac 180 agactgactg cctcatccac ttgtggcctg aatggccccc agccctactg catcgtcagt 240 cacctgcagg acgaaaagaa gtgcttcctt tgtgactccc ggcgcccctt ctctgctaga 300 gacaacccac acagccatcg catccagaat gtagtcacca gctttgcacc acagcggcgg 360 gcagcctggt ggcagtcaga gaatggtatc cctgcggtca ccatccagct ggacctggag 420 gctgagtttc atttcacaca cctcattatg accttcaaga catttcgccc tgctgccatg 480 ctggtggaac gctcagcaga ctttggccgc acctggcatg tgtaccgata tttctcctat 540 gactgtgggg ctgacttccc aggagtccca ctagcacccc cacggcactg ggatgatgta 600 gtctgtgagt cccgctactc agagattgag ccatccactg aaggcgaggt catctatcgt 660 gtgctggacc ctgccatccc tatcccagac ccctacagct cacggattca gaacctgttg 720 aagatcacca acctacgggt gaacctgact cgtctacaca cgttgggaga caacctactc 780 gacccacgga gggagatccg agagaagtac tactatgccc tctatgagct ggttgtacgt 840 ggcaactgct tctgctacgg acacgcctca gagtgtgcac ccgccccagg ggcaccagcc 900 catgctgagg gcatggtgca cggagcttgc atctgcaaac acaacacacg tggcctcaac 960 tgcgagcagt gtcaggattt ctatcgtgac ctgccctggc gtccggctga ggacggccat 1020 agtcatgcct gtaggaagtg tgagtgccat gggcacaccc acagctgcca cttcgacatg 1080 gccgtatacc tggcatctgg caatgtgagt ggaggtgtgt gtgatggatg tcagcataac 1140 acagctgggc gccactgtga gctctgtcgg cccttcttct accgtgaccc aaccaaggac 1200 ctgcgggatc cggctgtgtg ccgctcctgt gattgtgacc ccatgggttc tcaagacggt 1260 ggtcgctgtg attcccatga tgaccctgca ctgggactgg tctccggcca gtgtcgctgc 1320 aaagaacatg tggtgggcac tcgctgccag caatgccgtg atggcttctt tgggctcagc 1380 atcagtgacc gtctgggctg ccggcgatgt caatgtaatg cacggggcac agtgcctggg 1440 agcactcctt gtgaccccaa cagtggatcc tgttactgca aacgtctagt gactggacgt 1500 ggatgtgacc gctgcctgcc tggccactgg ggcctgagcc acgacctgct cggctgccgc 1560 ccctgtgact gcgacgtggg tggtgctttg gatccccagt gtgatgaggg cacaggtcaa 1620 tgccactgcc gccagcacat ggttgggcga cgctgtgagc aggtgcaacc tggctacttc 1680 cggcccttcc tggaccacct aatttgggag gctgaggaca cccgagggca ggtgctcgat 1740 gtggtggagc gcctggtgac ccccggggaa actccatcct ggactggctc aggcttcgtg 1800 cggctacagg aaggtcagac cctggagttc ctggtggcct ctgtgccgaa ggctatggac 1860 tatgacctgc tgctgcgctt agagccccag gtccctgagc aatgggcaga gttggaactg 1920 attgtgcagc gtccagggcc tgtgcctgcc cacagcctgt gtgggcattt ggtgcccaag 1980 gatgatcgca tccaagggac tctgcaacca catgccaggt acttgatatt tcctaatcct 2040 gtctgccttg agcctggtat ctcctacaag ctgcatctga agctggtacg gacaggggga 2100 agtgcccagc ctgagactcc ctactctgga cctggcctgc tcattgactc gctggtgctg 2160 ctgccccgtg tcctggtgct agagatgttt agtgggggtg atgctgctgc cctggagcgc 2220 caggccacct ttgaacgcta ccaatgccat gaggagggtc tggtgcccag caagacttct 2280 ccctctgagg cctgcgcacc cctcctcatc agcctgtcca ccctcatcta caatggtgcc 2340 ctgccatgtc agtgcaaccc tcaaggttca ctgagttctg agtgcaaccc tcatggtggt 2400 cagtgcctgt gcaagcctgg agtggttggg cgccgctgtg acctctgtgc ccctggctac 2460 tatggctttg gccccacagg ctgtcaagcc tgccagtgca gccacgaggg ggcactcagc 2520 agtctctgtg aaaagaccag tgggcaatgt ctctgtcgaa ctggtgcctt tgggcttcgc 2580 tgtgaccgct gccagcgtgg ccagtgggga ttccctagct gccggccatg tgtctgcaat 2640 gggcatgcag atgagtgcaa cacccacaca ggcgcttgcc tgggctgccg tgatcacaca 2700 gggggtgagc actgtgaaag gtgcattgct ggtttccacg gggacccacg gctgccatat 2760 gggggccagt gccggccctg tccctgtcct gaaggccctg ggagccaacg gcactttgct 2820 acttcttgcc accaggatga atattcccag cagattgtgt gccactgccg ggcaggctat 2880 acggggctgc gatgtgaagc ttgtgcccct gggcactttg gggacccatc aaggccaggt 2940 ggccggtgcc aactgtgtga gtgcagtggg aacattgacc caatggatcc tgatgcctgt 3000 gacccccaca cggggcaatg cctgcgctgt ttacaccaca cagagggtcc acactgtgcc 3060 cactgcaagc ctggcttcca tgggcaggct gcccgacaga gctgtcaccg ctgcacatgc 3120 aacctgctgg gcacaaatcc gcagcagtgc ccatctcctg accagtgcca ctgtgatcca 3180 agcagtgggc agtgcccatg cctccccaat gtccagggcc ctagctgtga ccgctgtgcc 3240 cccaacttct ggaacctcac cagtggccat ggttgccagc cttgtgcctg ccacccaagc 3300 cgggccagag gccccacctg caacgagttc acagggcagt gccactgccg tgccggcttt 3360 ggagggcgga cttgttctga gtgccaagag ctccactggg gagaccctgg gttgcagtgc 3420 catgcctgtg attgtgactc tcgtggaata gatacacctc agtgtcaccg cttcacaggt 3480 cactgcagct gccgcccagg ggtgtctggt gtgcgctgtg accagtgtgc ccgtggcttc 3540 tcaggaatct ttcctgcctg ccatccctgc catgcatgct tcggggattg ggaccgagtg 3600 gtgcaggact tggcagcccg tacacagcgc ctagagcagc gggcgcagga gttgcaacag 3660 acgggtgtgc tgggtgcctt tgagagcagc ttctggcaca tgcaggagaa gctgggcatt 3720 gtgcagggca tcgtaggtgc ccgcaacacc tcagccgcct ccactgcaca gcttgtggag 3780 gccacagagg agctgcggcg tgaaattggg gaggccactg agcacctgac tcagctcgag 3840 gcagacctga cagatgtgca agatgagaac ttcaatgcca accatgcact aagtggtctg 3900 gagcgagata ggcttgcact taatctcaca ctgcggcagc tcgaccagca tcttgacttg 3960 ctcaaacatt caaacttcct gggtgcctat gacagcatcc ggcatgccca tagccagtct 4020 gcagaggcag aacgtcgtgc caatacctca gccctggcag tacctagccc tgtgagcaac 4080 tcggcaagtg ctcggcatcg gacagaggca ctgatggatg ctcagaagga ggacttcaac 4140 agcaaacaca tggccaacca gcgggcactt ggcaagctct ctgcccatac ccacaccctg 4200 agcctgacag acataaatga gctggtgtgt ggggcaccag gggatgcacc ctgtgctaca 4260 agcccttgtg ggggtgccgg ctgtcgagat gaggatgggc agccgcgctg tgggggcctc 4320 agctgcaatg gggcagcggc tacagcagac ctagcactgg gccgggcccg gcacacacag 4380 gcagagctgc agcgggcact ggcagaaggt ggtagcatcc tcagcagagt ggctgagact 4440 cgtcggcagg caagcgaggc acagcagcgg gcccaggcag ccctggacaa ggctaatgct 4500 tccaggggac aggtggaaca ggccaaccag gaacttcaag aacttatcca gagtgtgaag 4560 gacttcctca accaggaggg ggctgatcct gatagcattg aaatggtggc cacacgggtg 4620 ctagagctct ccatcccagc ttcagctgag cagatccagc acctggcggg tgcgattgca 4680 gagcgagtcc ggagcctggc agatgtggat gcgatcctgg cacgtactgt aggagatgtg 4740 cgtcgtgccg agcagctact gcaggatgca cggcgggcaa ggagctgggc tgaggatgag 4800 aaacagaagg cagagacagt acaggcagca ctggaggagg cccagcgggc acagggtatt 4860 gcccagggtg ccatccgggg ggcagtggct gacacacggg acacagagca gaccctgtac 4920 caggtacagg agaggatggc aggtgcagag cgggcactga gctctgcagg tgaaagggct 4980 cggcagttgg atgctctcct ggaggctctg aaattgaaac gggcaggaaa tagtctggca 5040 gcctctacag cagaagaaac ggcaggcagt gcccagggtc gtgcccagga ggctgagcag 5100 ctgctacgcg gtcctctggg tgatcagtac cagacggtga aggccctagc tgagcgcaag 5160 gcccaaggtg tgctggctgc acaggcaagg gcagaacaac tgcgggatga ggctcgggac 5220 ctgttgcaag ccgctcagga caagctgcag cggctacagg aattggaagg cacctatgag 5280 gaaaatgagc gggcactgga gagtaaggca gcccagttgg acgggttgga ggccaggatg 5340 cgcagcgtgc ttcaagccat caacttgcag gtgcagatct acaacacctg ccagtga 5397 <210> 8 <211> 1609 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit gamma-1 (Homo sapiens) <400> 8 Met Arg Gly Ser His Arg Ala Ala Pro Ala Leu Arg Pro Arg Gly Arg   1 5 10 15 Leu Trp Pro Val Leu Ala Val Leu Ala Ala Ala Ala Ala Ala Gly Cys              20 25 30 Ala Gln Ala Ala Met Asp Glu Cys Thr Asp Glu Gly Gly Arg Pro Gln          35 40 45 Arg Cys Met Pro Glu Phe Val Asn Ala Ala Phe Asn Val Thr Val Val      50 55 60 Ala Thr Asn Thr Cys Gly Thr Pro Pro Glu Glu Tyr Cys Val Gln Thr  65 70 75 80 Gly Val Thr Gly Val Thr Lys Ser Cys His Leu Cys Asp Ala Gly Gln                  85 90 95 Pro His Leu Gln His Gly Ala Ala Phe Leu Thr Asp Tyr Asn Asn Gln             100 105 110 Ala Asp Thr Thr Trp Trp Gln Ser Gln Thr Met Leu Ala Gly Val Gln         115 120 125 Tyr Pro Ser Ser Ile Asn Leu Thr Leu His Leu Gly Lys Ala Phe Asp     130 135 140 Ile Thr Tyr Val Arg Leu Lys Phe His Thr Ser Arg Pro Glu Ser Phe 145 150 155 160 Ala Ile Tyr Lys Arg Thr Arg Glu Asp Gly Pro Trp Ile Pro Tyr Gln                 165 170 175 Tyr Tyr Ser Gly Ser Cys Glu Asn Thr Tyr Ser Lys Ala Asn Arg Gly             180 185 190 Phe Ile Arg Thr Gly Gly Asp Glu Gln Gln Ala Leu Cys Thr Asp Glu         195 200 205 Phe Ser Asp Ile Ser Pro Leu Thr Gly Gly Asn Val Ala Phe Ser Thr     210 215 220 Leu Glu Gly Arg Pro Ser Ala Tyr Asn Phe Asp Asn Ser Pro Val Leu 225 230 235 240 Gln Glu Trp Val Thr Ala Thr Asp Ile Arg Val Thr Leu Asn Arg Leu                 245 250 255 Asn Thr Phe Gly Asp Glu Val Phe Asn Asp Pro Lys Val Leu Lys Ser             260 265 270 Tyr Tyr Tyr Ala Ile Ser Asp Phe Ala Val Gly Gly Arg Cys Lys Cys         275 280 285 Asn Gly His Ala Ser Glu Cys Met Lys Asn Glu Phe Asp Lys Leu Val     290 295 300 Cys Asn Cys Lys His Asn Thr Tyr Gly Val Asp Cys Glu Lys Cys Leu 305 310 315 320 Pro Phe Phe Asn Asp Arg Pro Trp Arg Arg Ala Thr Ala Glu Ser Ala                 325 330 335 Ser Glu Cys Leu Pro Cys Asp Cys Asn Gly Arg Ser Gln Glu Cys Tyr             340 345 350 Phe Asp Pro Glu Leu Tyr Arg Ser Thr Gly His Gly Gly His Cys Thr         355 360 365 Asn Cys Gln Asp Asn Thr Asp Gly Ala His Cys Glu Arg Cys Arg Glu     370 375 380 Asn Phe Phe Arg Leu Gly Asn Asn Glu Ala Cys Ser Ser Cys His Cys 385 390 395 400 Ser Pro Val Gly Ser Leu Ser Thr Gln Cys Asp Ser Tyr Gly Arg Cys                 405 410 415 Ser Cys Lys Pro Gly Val Met Gly Asp Lys Cys Asp Arg Cys Gln Pro             420 425 430 Gly Phe His Ser Leu Thr Glu Ala Gly Cys Arg Pro Cys Ser Cys Asp         435 440 445 Pro Ser Gly Ser Ile Asp Glu Cys Asn Ile Glu Thr Gly Arg Cys Val     450 455 460 Cys Lys Asp Asn Val Glu Gly Phe Asn Cys Glu Arg Cys Lys Pro Gly 465 470 475 480 Phe Phe Asn Leu Glu Ser Ser Asn Pro Arg Gly Cys Thr Pro Cys Phe                 485 490 495 Cys Phe Gly His Ser Ser Val Cys Thr Asn Ala Val Gly Tyr Ser Val             500 505 510 Tyr Ser Ile Ser Ser Thr Phe Gln Ile Asp Glu Asp Gly Trp Arg Ala         515 520 525 Glu Gln Arg Asp Gly Ser Glu Ala Ser Leu Glu Trp Ser Ser Glu Arg     530 535 540 Gln Asp Ile Ala Val Ile Ser Asp Ser Tyr Phe Pro Arg Tyr Phe Ile 545 550 555 560 Ala Pro Ala Lys Phe Leu Gly Lys Gln Val Leu Ser Tyr Gly Gln Asn                 565 570 575 Leu Ser Phe Ser Phe Arg Val Asp Arg Arg Asp Thr Arg Leu Ser Ala             580 585 590 Glu Asp Leu Val Leu Glu Gly Ala Gly Leu Arg Val Ser Val Pro Leu         595 600 605 Ile Ala Gln Gly Asn Ser Tyr Pro Ser Glu Thr Thr Val Lys Tyr Val     610 615 620 Phe Arg Leu His Glu Ala Thr Asp Tyr Pro Trp Arg Pro Ala Leu Thr 625 630 635 640 Pro Phe Glu Phe Gln Lys Leu Leu Asn Asn Leu Thr Ser Ile Lys Ile                 645 650 655 Arg Gly Thr Tyr Ser Glu Arg Ser Ala Gly Tyr Leu Asp Asp Val Thr             660 665 670 Leu Ala Ser Ala Arg Pro Gly Pro Gly Val Pro Ala Thr Trp Val Glu         675 680 685 Ser Cys Thr Cys Pro Val Gly Tyr Gly Gly Gln Phe Cys Glu Met Cys     690 695 700 Leu Ser Gly Tyr Arg Arg Glu Thr Pro Asn Leu Gly Pro Tyr Ser Pro 705 710 715 720 Cys Val Leu Cys Ala Cys Asn Gly His Ser Glu Thr Cys Asp Pro Glu                 725 730 735 Thr Gly Val Cys Asn Cys Arg Asp Asn Thr Ala Gly Pro His Cys Glu             740 745 750 Lys Cys Ser Asp Gly Tyr Tyr Gly Asp Ser Thr Ala Gly Thr Ser Ser         755 760 765 Asp Cys Gln Pro Cys Pro Cys Pro Gly Gly Ser Ser Cys Ala Val Val     770 775 780 Pro Lys Thr Lys Glu Val Val Cys Thr Asn Cys Pro Thr Gly Thr Thr 785 790 795 800 Gly Lys Arg Cys Glu Leu Cys Asp Asp Gly Tyr Phe Gly Asp Pro Leu                 805 810 815 Gly Arg Asn Gly Pro Val Arg Leu Cys Arg Leu Cys Gln Cys Ser Asp             820 825 830 Asn Ile Asp Pro Asn Ala Val Gly Asn Cys Asn Arg Leu Thr Gly Glu         835 840 845 Cys Leu Lys Cys Ile Tyr Asn Thr Ala Gly Phe Tyr Cys Asp Arg Cys     850 855 860 Lys Asp Gly Phe Phe Gly Asn Pro Leu Ala Pro Asn Pro Ala Asp Lys 865 870 875 880 Cys Lys Ala Cys Asn Cys Asn Leu Tyr Gly Thr Met Lys Gln Gln Ser                 885 890 895 Ser Cys Asn Pro Val Thr Gly Gln Cys Glu Cys Leu Pro His Val Thr             900 905 910 Gly Gln Asp Cys Gly Ala Cys Asp Pro Gly Phe Tyr Asn Leu Gln Ser         915 920 925 Gly Gln Gly Cys Glu Arg Cys Asp Cys His Ala Leu Gly Ser Thr Asn     930 935 940 Gly Gln Cys Asp Ile Arg Thr Gly Gln Cys Glu Cys Gln Pro Gly Ile 945 950 955 960 Thr Gly Gln His Cys Glu Arg Cys Glu Val Asn His Phe Gly Phe Gly                 965 970 975 Pro Glu Gly Cys Lys Pro Cys Asp Cys His Pro Glu Gly Ser Leu Ser             980 985 990 Leu Gln Cys Lys Asp Asp Gly Arg Cys Glu Cys Arg Glu Gly Phe Val         995 1000 1005 Gly Asn Arg Cys Asp Gln Cys Glu Glu Asn Tyr Phe Tyr Asn Arg Ser    1010 1015 1020 Trp Pro Gly Cys Gln Glu Cys Pro Ala Cys Tyr Arg Leu Val Lys Asp 1025 1030 1035 1040 Lys Val Ala Asp His Arg Val Lys Leu Gln Glu Leu Glu Ser Leu Ile                1045 1050 1055 Ala Asn Leu Gly Thr Gly Asp Glu Met Val Thr Asp Gln Ala Phe Glu            1060 1065 1070 Asp Leg Leu Lys Glu Ala Glu Arg Glu Val Met Asp Leu Leu Arg Glu        1075 1080 1085 Ala Gln Asp Val Lys Asp Val Asp Gln Asn Leu Met Asp Arg Leu Gln    1090 1095 1100 Arg Val Asn Asn Thr Leu Ser Ser Gln Ile Ser Arg Leu Gln Asn Ile 1105 1110 1115 1120 Arg Asn Thr Ile Glu Glu Thr Gly Asn Leu Ala Glu Gln Ala Arg Ala                1125 1130 1135 His Val Glu Asn Thr Glu Arg Leu Ile Glu Ile Ala Ser Arg Glu Leu            1140 1145 1150 Glu Lys Ala Lys Val Ala Ala Ala Asn Val Ser Val Thr Gln Pro Glu        1155 1160 1165 Ser Thr Gly Asp Pro Asn Asn Met Thr Leu Leu Ala Glu Glu Ala Arg    1170 1175 1180 Lys Leu Ala Glu Arg His Lys Gln Glu Ala Asp Asp Ile Val Arg Val 1185 1190 1195 1200 Ala Lys Thr Ala Asn Asp Thr Ser Thr Glu Ala Tyr Asn Leu Leu Leu                1205 1210 1215 Arg Thr Leu Ala Gly Glu Asn Gln Thr Ala Phe Glu Ile Glu Glu Leu            1220 1225 1230 Asn Arg Lys Tyr Glu Gln Ala Lys Asn Ile Ser Gln Asp Leu Glu Lys        1235 1240 1245 Gln Ala Ala Arg Val His Glu Glu Ala Lys Arg Ala Gly Asp Lys Ala    1250 1255 1260 Val Glu Ile Tyr Ala Ser Val Ala Gln Leu Ser Pro Leu Asp Ser Glu 1265 1270 1275 1280 Thr Leu Glu Asn Glu Ala Asn Asn Ile Lys Met Glu Ala Glu Asn Leu                1285 1290 1295 Glu Gln Leu Ile Asp Gln Lys Leu Lys Asp Tyr Glu Asp Leu Arg Glu            1300 1305 1310 Asp Met Arg Gly Lys Glu Leu Glu Val Lys Asn Leu Leu Glu Lys Gly        1315 1320 1325 Lys Thr Glu Gln Gln Thr Ala Asp Gln Leu Leu Ala Arg Ala Asp Ala    1330 1335 1340 Ala Lys Ala Leu Ala Glu Glu Ala Ala Lys Lys Gly Arg Asp Thr Leu 1345 1350 1355 1360 Gln Glu Ala Asn Asp Ile Leu Asn Asn Leu Lys Asp Phe Asp Arg Arg                1365 1370 1375 Val Asn Asp Asn Lys Thr Ala Ala Glu Glu Ala Leu Arg Lys Ile Pro            1380 1385 1390 Ala Ile Asn Gln Thr Ile Thr Glu Ala Asn Glu Lys Thr Arg Glu Ala        1395 1400 1405 Gln Gln Ala Leu Gly Ser Ala Ala Ala Asp Ala Thr Glu Ala Lys Asn    1410 1415 1420 Lys Ala His Glu Ala Glu Arg Ile Ala Ser Ala Val Gln Lys Asn Ala 1425 1430 1435 1440 Thr Ser Thr Lys Ala Glu Ala Glu Arg Thr Phe Ala Glu Val Thr Asp                1445 1450 1455 Leu Asp Asn Glu Val Asn Asn Met Leu Lys Gln Leu Gln Glu Ala Glu            1460 1465 1470 Lys Glu Leu Lys Arg Lys Gln Asp Asp Ala Asp Gln Asp Met Met Met        1475 1480 1485 Ala Gly Met Ala Ser Gln Ala Ala Gln Glu Ala Glu Ile Asn Ala Arg    1490 1495 1500 Lys Ala Lys Asn Ser Val Thr Ser Leu Leu Ser Ile Ile Asn Asp Leu 1505 1510 1515 1520 Leu Glu Gln Leu Gly Gln Leu Asp Thr Val Asp Leu Asn Lys Leu Asn                1525 1530 1535 Glu Ile Glu Gly Thr Leu Asn Lys Ala Lys Asp Glu Met Lys Val Ser            1540 1545 1550 Asp Leu Asp Arg Lys Val Ser Asp Leu Glu Asn Glu Ala Lys Lys Gln        1555 1560 1565 Glu Ala Ala Ile Met Asp Tyr Asn Arg Asp Ile Glu Glu Ile Met Lys    1570 1575 1580 Asp Ile Arg Asn Leu Glu Asp Ile Arg Lys Thr Leu Pro Ser Gly Cys 1585 1590 1595 1600 Phe Asn Thr Pro Ser Ile Glu Lys Pro                1605 <210> 9 <211> 4830 <212> DNA <213> Artificial Sequence <220> <223> DNA sequence of Laminin subunit gamma-1 (Homo sapiens) <400> 9 atgagaggga gccatcgggc cgcgccggcc ctgcggcccc gggggcggct ctggcccgtg 60 ctggccgtgc tggcggcggc cgccgcggcg ggctgtgccc aggcagccat ggacgagtgc 120 acggacgagg gcgggcggcc gcagcgctgc atgcccgagt tcgtcaacgc cgccttcaac 180 gtgactgtgg tggccaccaa cacgtgtggg actccgcccg aggaatactg tgtgcagacc 240 ggggtgaccg gggtcaccaa gtcctgtcac ctgtgcgacg ccgggcagcc ccacctgcag 300 cacggggcag ccttcctgac cgactacaac aaccaggccg acaccacctg gtggcaaagc 360 cagaccatgc tggccggggt gcagtacccc agctccatca acctcacgct gcacctggga 420 aaagcttttg acatcaccta tgtgcgtctc aagttccaca ccagccgccc ggagagcttt 480 gccatttaca agcgcacacg ggaagacggg ccctggattc cttaccagta ctacagtggt 540 tcctgtgaga acacctactc caaggcaaac cgcggcttca tcaggacagg aggggacgag 600 cagcaggcct tgtgtactga tgaattcagt gacatttctc ccctcactgg gggcaacgtg 660 gccttttcta ccctggaagg aaggcccagc gcctataact ttgacaatag ccctgtgctg 720 caggaatggg taactgccac tgacatcaga gtaactctta atcgcctgaa cacttttgga 780 gatgaagtgt ttaacgatcc caaagttctc aagtcctatt attatgccat ctctgatttt 840 gctgtaggtg gcagatgtaa atgtaatgga cacgcaagcg agtgtatgaa gaacgaattt 900 gataagctgg tgtgtaattg caaacataac acatatggag tagactgtga aaagtgtctt 960 cctttcttca atgaccggcc gtggaggagg gcaactgcgg aaagtgccag tgaatgcctg 1020 ccctgtgatt gcaatggtcg atcccaggaa tgctacttcg accctgaact ctatcgttcc 1080 actggccatg ggggccactg taccaactgc caggataaca cagatggcgc ccactgtgag 1140 aggtgccgag agaacttctt ccgccttggc aacaatgaag cctgctcttc atgccactgt 1200 agtcctgtgg gctctctaag cacacagtgt gatagttacg gcagatgcag ctgtaagcca 1260 ggagtgatgg gggacaaatg tgaccgttgc cagcctggat tccattctct cactgaagca 1320 ggatgcaggc catgctcttg tgatccctct ggcagcatag atgaatgtaa tattgaaaca 1380 ggaagatgtg tttgcaaaga caatgtcgaa ggcttcaatt gtgaaagatg caaacctgga 1440 ttttttaatc tggaatcatc taatcctcgg ggttgcacac cctgcttctg ctttgggcat 1500 tcttctgtct gtacaaacgc tgttggctac agtgtttatt ctatctcctc tacctttcag 1560 attgatgagg atgggtggcg tgcggaacag agagatggct ctgaagcatc tctcgagtgg 1620 tcctctgaga ggcaagatat cgccgtgatc tcagacagct actttcctcg gtacttcatt 1680 gctcctgcaa agttcttggg caagcaggtg ttgagttatg gtcagaacct ctccttctcc 1740 tttcgagtgg acaggcgaga tactcgcctc tctgcagaag accttgtgct tgagggagct 1800 ggcttaagag tatctgtacc cttgatcgct cagggcaatt cctatccaag tgagaccact 1860 gtgaagtatg tcttcaggct ccatgaagca acagattacc cttggaggcc tgctcttacc 1920 ccttttgaat ttcagaagct cctaaacaac ttgacctcta tcaagatacg tgggacatac 1980 agtgagagaa gtgctggata tttggatgat gtcaccctgg caagtgctcg tcctgggcct 2040 ggagtccctg caacttgggt ggagtcctgc acctgtcctg tgggatatgg agggcagttt 2100 tgtgagatgt gcctctcagg ttacagaaga gaaactccta atcttggacc atacagtcca 2160 tgtgtgcttt gcgcctgcaa tggacacagc gagacctgtg atcctgagac aggtgtttgt 2220 aactgcagag acaatacggc tggcccgcac tgtgagaagt gcagtgatgg gtactatgga 2280 gattcaactg caggcacctc ctccgattgc caaccctgtc cgtgtcctgg aggttcaagt 2340 tgtgctgttg ttcccaagac aaaggaggtg gtgtgcacca actgtcctac tggcaccact 2400 ggtaagagat gtgagctctg tgatgatggc tactttggag accccctggg tagaaacggc 2460 cctgtgagac tttgccgcct gtgccagtgc agtgacaaca tcgatcccaa tgcagttgga 2520 aattgcaatc gcttgacggg agaatgcctg aagtgcatct ataacactgc tggcttctat 2580 tgtgaccggt gcaaagacgg attttttgga aatcccctgg ctcccaatcc agcagacaaa 2640 tgcaaagcct gcaattgcaa tctgtatggg accatgaagc agcagagcag ctgtaacccc 2700 gtgacggggc agtgtgaatg tttgcctcac gtgactggcc aggactgtgg tgcttgtgac 2760 cctggattct acaatctgca gagtgggcaa ggctgtgaga ggtgtgactg ccatgccttg 2820 ggctccacca atgggcagtg tgacatccgc accggccagt gtgagtgcca gcccggcatc 2880 actggtcagc actgtgagcg ctgtgaggtc aaccactttg ggtttggacc tgaaggctgc 2940 aaaccctgtg actgtcatcc tgagggatct ctttcacttc agtgcaaaga tgatggtcgc 3000 tgtgaatgca gagaaggctt tgtgggaaat cgctgtgacc agtgtgaaga aaactatttc 3060 tacaatcggt cttggcctgg ctgccaggaa tgtccagctt gttaccggct ggtaaaggat 3120 aaggttgctg atcatagagt gaagctccag gaattagaga gtctcatagc aaaccttgga 3180 actggggatg agatggtgac agatcaagcc ttcgaggata gactaaagga agcagagagg 3240 gaagttatgg acctccttcg tgaggcccag gatgtcaaag atgttgacca gaatttgatg 3300 gatcgcctac agagagtgaa taacactctg tccagccaaa ttagccgttt acagaatatc 3360 cggaatacca ttgaagagac tggaaacttg gctgaacaag cgcgtgccca tgtagagaac 3420 acagagcggt tgattgaaat cgcatccaga gaacttgaga aagcaaaagt cgctgctgcc 3480 aatgtgtcag tcactcagcc agaatctaca ggggacccaa acaacatgac tcttttggca 3540 gaagaggctc gaaagcttgc tgaacgtcat aaacaggaag ctgatgacat tgttcgagtg 3600 gcaaagacag ccaatgatac gtcaactgag gcatacaacc tgcttctgag gacactggca 3660 ggagaaaatc aaacagcatt tgagattgaa gagcttaata ggaagtatga acaagcgaag 3720 aacatctcac aggatctgga aaaacaagct gcccgagtac atgaggaggc caaaagggcc 3780 ggtgacaaag ctgtggagat ctatgccagc gtggctcagc tgagcccttt ggactctgag 3840 acactggaga atgaagcaaa taacataaag atggaagctg agaatctgga acaactgatt 3900 gaccagaaat taaaagatta tgaggacctc agagaagata tgagagggaa ggaacttgaa 3960 gtcaagaacc ttctggagaa aggcaagact gaacagcaga ccgcagacca actcctagcc 4020 cgagctgatg ctgccaaggc cctcgctgaa gaagctgcaa agaagggacg ggatacctta 4080 caagaagcta atgacattct caacaacctg aaagattttg ataggcgtgt gaacgataac 4140 aagacggccg cagaggaggc actaaggaag attcctgcca tcaaccagac catcactgaa 4200 gccaatgaaa agaccagaga agcccagcag gccctgggca gtgctgcggc ggatgccaca 4260 gaggccaaga acaaggccca tgaggcggag aggatcgcga gcgctgtcca aaagaatgcc 4320 accagcacca aggcagaagc tgaaagaact tttgcagaag ttacagatct ggataatgag 4380 gtgaacaata tgttgaagca actgcaggaa gcagaaaaag agctaaagag aaaacaagat 4440 gacgctgacc aggacatgat gatggcaggg atggcttcac aggctgctca agaagccgag 4500 atcaatgcca gaaaagccaa aaactctgtt actagcctcc tcagcattat taatgacctc 4560 ttggagcagc tggggcagct ggatacagtg gacctgaata agctaaacga gattgaaggc 4620 accctaaaca aagccaaaga tgaaatgaag gtcagcgatc ttgataggaa agtgtctgac 4680 ctggagaatg aagccaagaa gcaggaggct gccatcatgg actataaccg agatatcgag 4740 gagatcatga aggacattcg caatctggag gacatcagga agaccttacc atctggctgc 4800 ttcaacaccc cgtccattga aaagccctag 4830 <210> 10 <211> 3122 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit alpha-2 (Homo sapiens) <400> 10 Met Pro Gly Ala Ala Gly Val Leu Leu Leu Leu Leu Leu Ser Gly Gly   1 5 10 15 Leu Gly Gly Val Gln Ala Gln Arg Pro Gln Gln Gln Arg Gln Ser Gln              20 25 30 Ala His Gln Gln Arg Gly Leu Phe Pro Ala Val Leu Asn Leu Ala Ser          35 40 45 Asn Ala Leu Ile Thr Thr Asn Ala Thr Cys Gly Glu Lys Gly Pro Glu      50 55 60 Met Tyr Cys Lys Leu Val Glu His Val Pro Gly Gln Pro Val Arg Asn  65 70 75 80 Pro Gln Cys Arg Ile Cys Asn Gln Asn Ser Ser Asn Pro Asn Gln Arg                  85 90 95 His Pro Ile Thr Asn Ala Ile Asp Gly Lys Asn Thr Trp Trp Gln Ser             100 105 110 Pro Ser Ile Lys Asn Gly Ile Glu Tyr His Tyr Val Thr Ile Thr Leu         115 120 125 Asp Leu Gln Gln Val Phe Gln Ile Ala Tyr Val Ile Val Lys Ala Ala     130 135 140 Asn Ser Pro Arg Pro Gly Asn Trp Ile Leu Glu Arg Ser Leu Asp Asp 145 150 155 160 Val Glu Tyr Lys Pro Trp Gln Tyr His Ala Val Thr Asp Thr Glu Cys                 165 170 175 Leu Thr Leu Tyr Asn Ile Tyr Pro Arg Thr Gly Pro Pro Ser Tyr Ala             180 185 190 Lys Asp Asp Glu Val Ile Cys Thr Ser Phe Tyr Ser Lys Ile His Pro         195 200 205 Leu Glu Asn Gly Glu Ile His Ile Ser Leu Ile Asn Gly Arg Pro Ser     210 215 220 Ala Asp Asp Pro Ser Pro Glu Leu Leu Glu Phe Thr Ser Ala Arg Tyr 225 230 235 240 Ile Arg Leu Arg Phe Gln Arg Ile Arg Thr Leu Asn Ala Asp Leu Met                 245 250 255 Met Phe Ala His Lys Asp Pro Arg Glu Ile Asp Pro Ile Val Thr Arg             260 265 270 Arg Tyr Tyr Tyr Ser Val Lys Asp Ile Ser Val Gly Gly Met Cys Ile         275 280 285 Cys Tyr Gly His Ala Arg Ala Cys Pro Leu Asp Pro Ala Thr Asn Lys     290 295 300 Ser Arg Cys Glu Cys Glu His Asn Thr Cys Gly Asp Ser Cys Asp Gln 305 310 315 320 Cys Cys Pro Gly Phe His Gln Lys Pro Trp Arg Ala Gly Thr Phe Leu                 325 330 335 Thr Lys Thr Glu Cys Glu Ala Cys Asn Cys His Gly Lys Ala Glu Glu             340 345 350 Cys Tyr Tyr Asp Glu Asn Val Ala Arg Arg Asn Leu Ser Leu Asn Ile         355 360 365 Arg Gly Lys Tyr Ile Gly Gly Gly Val Cys Ile Asn Cys Thr Gln Asn     370 375 380 Thr Ala Gly Ile Asn Cys Glu Thr Cys Thr Asp Gly Phe Phe Arg Pro 385 390 395 400 Lys Gly Val Ser Pro Asn Tyr Pro Arg Pro Cys Gln Pro Cys His Cys                 405 410 415 Asp Pro Ile Gly Ser Leu Asn Glu Val Cys Val Lys Asp Glu Lys His             420 425 430 Ala Arg Arg Gly Leu Ala Pro Gly Ser Cys His Cys Lys Thr Gly Phe         435 440 445 Gly Gly Val Ser Cys Asp Arg Cys Ala Arg Gly Tyr Thr Gly Tyr Pro     450 455 460 Asp Cys Lys Ala Cys Asn Cys Ser Gly Leu Gly Ser Lys Asn Glu Asp 465 470 475 480 Pro Cys Phe Gly Pro Cys Ile Cys Lys Glu Asn Val Glu Gly Gly Asp                 485 490 495 Cys Ser Arg Cys Lys Ser Gly Phe Phe Asn Leu Gln Glu Asp Asn Trp             500 505 510 Lys Gly Cys Asp Glu Cys Phe Cys Ser Gly Val Ser Asn Arg Cys Gln         515 520 525 Ser Ser Tyr Trp Thr Tyr Gly Lys Ile Gln Asp Met Ser Gly Trp Tyr     530 535 540 Leu Thr Asp Leu Pro Gly Arg Ile Arg Val Ala Pro Gln Gln Asp Asp 545 550 555 560 Leu Asp Ser Pro Gln Gln Ile Ser Ile Ser Asn Ala Glu Ala Arg Gln                 565 570 575 Ala Leu Pro His Ser Tyr Tyr Trp Ser Ala Pro Ala Pro Tyr Leu Gly             580 585 590 Asn Lys Leu Pro Ala Val Gly Gly Gln Leu Thr Phe Thr Ile Ser Tyr         595 600 605 Asp Leu Glu Glu Glu Glu Glu Asp Thr Glu Arg Val Leu Gln Leu Met     610 615 620 Ile Ile Leu Glu Gly Asn Asp Leu Ser Ile Ser Thr Ala Gln Asp Glu 625 630 635 640 Val Tyr Leu His Pro Ser Glu Glu His Thr Asn Val Leu Leu Leu Lys                 645 650 655 Glu Glu Ser Phe Thr Ile His Gly Thr His Phe Pro Val Arg Arg Lys             660 665 670 Glu Phe Met Thr Val Leu Ala Asn Leu Lys Arg Val Leu Leu Gln Ile         675 680 685 Thr Tyr Ser Phe Gly Met Asp Ala Ile Phe Arg Leu Ser Ser Val Asn     690 695 700 Leu Glu Ser Ala Val Ser Tyr Pro Thr Asp Gly Ser Ile Ala Ala Ala 705 710 715 720 Val Glu Val Cys Gln Cys Pro Pro Gly Tyr Thr Gly Ser Ser Cys Glu                 725 730 735 Ser Cys Trp Pro Arg His Arg Arg Val Asn Gly Thr Ile Phe Gly Gly             740 745 750 Ile Cys Glu Pro Cys Gln Cys Phe Gly His Ala Glu Ser Cys Asp Asp         755 760 765 Val Thr Gly Glu Cys Leu Asn Cys Lys Asp His Thr Gly Gly Pro Tyr     770 775 780 Cys Asp Lys Cys Leu Pro Gly Phe Tyr Gly Glu Pro Thr Lys Gly Thr 785 790 795 800 Ser Glu Asp Cys Gln Pro Cys Ala Cys Pro Leu Asn Ile Pro Ser Asn                 805 810 815 Asn Phe Ser Pro Thr Cys His Leu Asp Arg Ser Leu Gly Leu Ile Cys             820 825 830 Asp Gly Cys Pro Val Gly Tyr Thr Gly Pro Arg Cys Glu Arg Cys Ala         835 840 845 Glu Gly Tyr Phe Gly Gln Pro Ser Val Pro Gly Gly Ser Cys Gln Pro     850 855 860 Cys Gln Cys Asn Asp Asn Leu Asp Phe Ser Ile Pro Gly Ser Cys Asp 865 870 875 880 Ser Leu Ser Gly Ser Cys Leu Ile Cys Lys Pro Gly Thr Thr Gly Arg                 885 890 895 Tyr Cys Glu Leu Cys Ala Asp Gly Tyr Phe Gly Asp Ala Val Asp Ala             900 905 910 Lys Asn Cys Gln Pro Cys Arg Cys Asn Ala Gly Gly Ser Phe Ser Glu         915 920 925 Val Cys His Ser Gln Thr Gly Gln Cys Glu Cys Arg Ala Asn Val Gln     930 935 940 Gly Gln Arg Cys Asp Lys Cys Lys Ala Gly Thr Phe Gly Leu Gln Ser 945 950 955 960 Ala Arg Gly Cys Val Pro Cys Asn Cys Asn Ser Phe Gly Ser Lys Ser                 965 970 975 Phe Asp Cys Glu Glu Ser Gly Gln Cys Trp Cys Gln Pro Gly Val Thr             980 985 990 Gly Lys Lys Cys Asp Arg Cys Ala His Gly Tyr Phe Asn Phe Gln Glu         995 1000 1005 Gly Gly Cys Thr Ala Cys Glu Cys Ser His Leu Gly Asn Asn Cys Asp    1010 1015 1020 Pro Lys Thr Gly Arg Cys Ile Cys Pro Pro Asn Thr Ile Gly Glu Lys 1025 1030 1035 1040 Cys Ser Lys Cys Ala Pro Asn Thr Trp Gly His Ser Ile Thr Thr Gly                1045 1050 1055 Cys Lys Ala Cys Asn Cys Ser Thr Val Gly Ser Leu Asp Phe Gln Cys            1060 1065 1070 Asn Val Asn Thr Gly Gln Cys Asn Cys His Pro Lys Phe Ser Gly Ala        1075 1080 1085 Lys Cys Thr Glu Cys Ser Arg Gly His Trp Asn Tyr Pro Arg Cys Asn    1090 1095 1100 Leu Cys Asp Cys Phe Leu Pro Gly Thr Asp Ala Thr Thr Cys Asp Ser 1105 1110 1115 1120 Glu Thr Lys Lys Cys Ser Cys Ser Asp Gln Thr Gly Gln Cys Thr Cys                1125 1130 1135 Lys Val Asn Val Glu Gly Ile His Cys Asp Arg Cys Arg Pro Gly Lys            1140 1145 1150 Phe Gly Leu Asp Ala Lys Asn Pro Leu Gly Cys Ser Ser Cys Tyr Cys        1155 1160 1165 Phe Gly Thr Thr Thr Gln Cys Ser Glu Ala Lys Gly Leu Ile Arg Thr    1170 1175 1180 Trp Val Thr Leu Lys Ala Glu Gln Thr Ile Leu Pro Leu Val Asp Glu 1185 1190 1195 1200 Ala Leu Gln His Thr Thr Thr Lys Gly Ile Val Phe Gln His Pro Glu                1205 1210 1215 Ile Val Ala His Met Asp Leu Met Arg Glu Asp Leu His Leu Glu Pro            1220 1225 1230 Phe Tyr Trp Lys Leu Pro Glu Gln Phe Glu Gly Lys Lys Leu Met Ala        1235 1240 1245 Tyr Gly Gly Lys Leu Lys Tyr Ala Ile Tyr Phe Glu Ala Arg Glu Glu    1250 1255 1260 Thr Gly Phe Ser Thr Tyr Asn Pro Gln Val Ile Ile Arg Gly Gly Thr 1265 1270 1275 1280 Pro Thr His Ala Arg Ile Ile Val Arg His Met Ala Ala Pro Leu Ile                1285 1290 1295 Gly Gln Leu Thr Arg His Glu Ile Glu Met Thr Glu Lys Glu Trp Lys            1300 1305 1310 Tyr Tyr Gly Asp Asp Pro Arg Val His Arg Thr Val Thr Arg Glu Asp        1315 1320 1325 Phe Leu Asp Ile Leu Tyr Asp Ile His Tyr Ile Leu Ile Lys Ala Thr    1330 1335 1340 Tyr Gly Asn Phe Met Arg Gln Ser Arg Ile Ser Glu Ile Ser Met Glu 1345 1350 1355 1360 Val Ala Glu Gln Gly Arg Gly Thr Thr Met Met Pro Pro Ala Asp Leu                1365 1370 1375 Ile Glu Lys Cys Asp Cys Pro Leu Gly Tyr Ser Gly Leu Ser Cys Glu            1380 1385 1390 Ala Cys Leu Pro Gly Phe Tyr Arg Leu Arg Ser Gln Pro Gly Gly Arg        1395 1400 1405 Thr Pro Gly Pro Thr Leu Gly Thr Cys Val Pro Cys Gln Cys Asn Gly    1410 1415 1420 His Ser Ser Leu Cys Asp Pro Glu Thr Ser Ile Cys Gln Asn Cys Gln 1425 1430 1435 1440 His His Thr Ala Gly Asp Phe Cys Glu Arg Cys Ala Leu Gly Tyr Tyr                1445 1450 1455 Gly Ile Val Lys Gly Leu Pro Asn Asp Cys Gln Gln Cys Ala Cys Pro            1460 1465 1470 Leu Ile Ser Ser Ser Asn Asn Phe Ser Pro Ser Cys Val Ala Glu Gly        1475 1480 1485 Leu Asp Asp Tyr Arg Cys Thr Ala Cys Pro Arg Gly Tyr Glu Gly Gln    1490 1495 1500 Tyr Cys Glu Arg Cys Ala Pro Gly Tyr Thr Gly Ser Pro Gly Asn Pro 1505 1510 1515 1520 Gly Gly Ser Cys Gln Glu Cys Glu Cys Asp Pro Tyr Gly Ser Leu Pro                1525 1530 1535 Val Pro Cys Asp Pro Val Thr Gly Phe Cys Thr Cys Arg Pro Gly Ala            1540 1545 1550 Thr Gly Arg Lys Cys Asp Gly Cys Lys His Trp His Ala Arg Glu Gly        1555 1560 1565 Trp Glu Cys Val Phe Cys Gly Asp Glu Cys Thr Gly Leu Leu Leu Gly    1570 1575 1580 Asp Leu Ala Arg Leu Glu Gln Met Val Met Ser Ile Asn Leu Thr Gly 1585 1590 1595 1600 Pro Leu Pro Ala Pro Tyr Lys Met Leu Tyr Gly Leu Glu Asn Met Thr                1605 1610 1615 Gln Glu Leu Lys His Leu Leu Ser Pro Gln Arg Ala Pro Glu Arg Leu            1620 1625 1630 Ile Gln Leu Ala Glu Gly Asn Leu Asn Thr Leu Val Thr Glu Met Asn        1635 1640 1645 Glu Leu Leu Thr Arg Ala Thr Lys Val Thr Ala Asp Gly Glu Gln Thr    1650 1655 1660 Gly Gln Asp Ala Glu Arg Thr Asn Thr Arg Ala Lys Ser Leu Gly Glu 1665 1670 1675 1680 Phe Ile Lys Glu Leu Ala Arg Asp Ala Glu Ala Val Asn Glu Lys Ala                1685 1690 1695 Ile Lys Leu Asn Glu Thr Leu Gly Thr Arg Asp Glu Ala Phe Glu Arg            1700 1705 1710 Asn Leu Glu Gly Leu Gln Lys Glu Ile Asp Gln Met Ile Lys Glu Leu        1715 1720 1725 Arg Arg Lys Asn Leu Glu Thr Gln Lys Glu Ile Ala Glu Asp Glu Leu    1730 1735 1740 Val Ala Ala Glu Ala Leu Leu Lys Lys Val Lys Lys Leu Phe Gly Glu 1745 1750 1755 1760 Ser Arg Gly Glu Asn Glu Glu Met Glu Lys Asp Leu Arg Glu Lys Leu                1765 1770 1775 Ala Asp Tyr Lys Asn Lys Val Asp Asp Ala Trp Asp Leu Leu Arg Glu            1780 1785 1790 Ala Thr Asp Lys Ile Arg Glu Ala Asn Arg Leu Phe Ala Val Asn Gln        1795 1800 1805 Lys Asn Met Thr Ala Leu Glu Lys Lys Lys Glu Ala Val Glu Ser Gly    1810 1815 1820 Lys Arg Gln Ile Glu Asn Thr Leu Lys Glu Gly Asn Asp Ile Leu Asp 1825 1830 1835 1840 Glu Ala Asn Arg Leu Ala Asp Glu Ile Asn Ser Ile Ile Asp Tyr Val                1845 1850 1855 Glu Asp Ile Gln Thr Lys Leu Pro Pro Met Ser Glu Glu Leu Asn Asp            1860 1865 1870 Lys Ile Asp Asp Leu Ser Gln Glu Ile Lys Asp Arg Lys Leu Ala Glu        1875 1880 1885 Lys Val Ser Gln Ala Glu Ser His Ala Ala Gln Leu Asn Asp Ser Ser    1890 1895 1900 Ala Val Leu Asp Gly Ile Leu Asp Glu Ala Lys Asn Ile Ser Phe Asn 1905 1910 1915 1920 Ala Thr Ala Ala Phe Lys Ala Tyr Ser Asn Ile Lys Asp Tyr Ile Asp                1925 1930 1935 Glu Ala Glu Lys Val Ala Lys Glu Ala Lys Asp Leu Ala His Glu Ala            1940 1945 1950 Thr Lys Leu Ala Thr Gly Pro Arg Gly Leu Leu Lys Glu Asp Ala Lys        1955 1960 1965 Gly Cys Leu Gln Lys Ser Phe Arg Ile Leu Asn Glu Ala Lys Lys Leu    1970 1975 1980 Ala Asn Asp Val Lys Glu Asn Glu Asp His Leu Asn Gly Leu Lys Thr 1985 1990 1995 2000 Arg Ile Glu Asn Ala Asp Ala Arg Asn Gly Asp Leu Leu Arg Thr Leu                2005 2010 2015 Asn Asp Thr Leu Gly Lys Leu Ser Ala Ile Pro Asn Asp Thr Ala Ala            2020 2025 2030 Lys Leu Gln Ala Val Lys Asp Lys Ala Arg Gln Ala Asn Asp Thr Ala        2035 2040 2045 Lys Asp Val Leu Ala Gln Ile Thr Glu Leu His Gln Asn Leu Asp Gly    2050 2055 2060 Leu Lys Lys Asn Tyr Asn Lys Leu Ala Asp Ser Val Ala Lys Thr Asn 2065 2070 2075 2080 Ala Val Val Lys Asp Pro Ser Lys Asn Lys Ile Ile Ala Asp Ala Asp                2085 2090 2095 Ala Thr Val Lys Asn Leu Glu Gln Glu Ala Asp Arg Leu Ile Asp Lys            2100 2105 2110 Leu Lys Pro Ile Lys Glu Leu Glu Asp Asn Leu Lys Lys Asn Ile Ser        2115 2120 2125 Glu Ile Lys Glu Leu Ile Asn Gln Ala Arg Lys Gln Ala Asn Ser Ile    2130 2135 2140 Lys Val Ser Val Ser Ser Gly Gly Asp Cys Ile Arg Thr Tyr Lys Pro 2145 2150 2155 2160 Glu Ile Lys Lys Gly Ser Tyr Asn Asn Ile Val Val Asn Val Lys Thr                2165 2170 2175 Ala Val Ala Asp Asn Leu Leu Phe Tyr Leu Gly Ser Ala Lys Phe Ile            2180 2185 2190 Asp Phe Leu Ala Ile Glu Met Arg Lys Gly Lys Val Ser Phe Leu Trp        2195 2200 2205 Asp Val Gly Ser Gly Val Gly Arg Val Glu Tyr Pro Asp Leu Thr Ile    2210 2215 2220 Asp Asp Ser Tyr Trp Tyr Arg Ile Val Ala Ser Arg Thr Gly Arg Asn 2225 2230 2235 2240 Gly Thr Ile Ser Val Arg Ala Leu Asp Gly Pro Lys Ala Ser Ile Val                2245 2250 2255 Pro Ser Thr His His Ser Thr Ser Pro Pro Gly Tyr Thr Ile Leu Asp            2260 2265 2270 Val Asp Ala Asn Ala Met Leu Phe Val Gly Gly Leu Thr Gly Lys Leu        2275 2280 2285 Lys Lys Ala Asp Ala Val Arg Val Ile Thr Phe Thr Gly Cys Met Gly    2290 2295 2300 Glu Thr Tyr Phe Asp Asn Lys Pro Ile Gly Leu Trp Asn Phe Arg Glu 2305 2310 2315 2320 Lys Glu Gly Asp Cys Lys Gly Cys Thr Val Ser Pro Gln Val Glu Asp                2325 2330 2335 Ser Glu Gly Thr Ile Gln Phe Asp Gly Glu Gly Tyr Ala Leu Val Ser            2340 2345 2350 Arg Pro Ile Arg Trp Tyr Pro Asn Ile Ser Thr Val Met Phe Lys Phe        2355 2360 2365 Arg Thr Phe Ser Ser Ser Ala Leu Leu Met Tyr Leu Ala Thr Arg Asp    2370 2375 2380 Leu Arg Asp Phe Met Ser Val Glu Leu Thr Asp Gly His Ile Lys Val 2385 2390 2395 2400 Ser Tyr Asp Leu Gly Ser Gly Met Ala Ser Val Val Ser Asn Gln Asn                2405 2410 2415 His Asn Asp Gly Lys Trp Lys Ser Phe Thr Leu Ser Arg Ile Gln Lys            2420 2425 2430 Gln Ala Asn Ile Ser Ile Val Asp Ile Asp Thr Asn Gln Glu Glu Asn        2435 2440 2445 Ile Ala Thr Ser Ser Ser Gly Asn Asn Phe Gly Leu Asp Leu Lys Ala    2450 2455 2460 Asp Asp Lys Ile Tyr Phe Gly Gly Leu Pro Thr Leu Arg Asn Leu Ser 2465 2470 2475 2480 Met Lys Ala Arg Pro Glu Val Asn Leu Lys Lys Tyr Ser Gly Cys Leu                2485 2490 2495 Lys Asp Ile Glu Ile Ser Arg Thr Pro Tyr Asn Ile Leu Ser Ser Pro            2500 2505 2510 Asp Tyr Val Gly Val Thr Lys Gly Cys Ser Leu Glu Asn Val Tyr Thr        2515 2520 2525 Val Ser Phe Pro Lys Pro Gly Phe Val Glu Leu Ser Pro Val Pro Ile    2530 2535 2540 Asp Val Gly Thr Glu Ile Asn Leu Ser Phe Ser Thr Lys Asn Glu Ser 2545 2550 2555 2560 Gly Ile Ile Leu Leu Gly Ser Gly Gly Thr Pro Ala Pro Pro Arg Arg                2565 2570 2575 Lys Arg Arg Gln Thr Gly Gln Ala Tyr Tyr Ala Ile Leu Leu Asn Arg            2580 2585 2590 Gly Arg Leu Glu Val His Leu Ser Thr Gly Ala Arg Thr Met Arg Lys        2595 2600 2605 Ile Val Ile Arg Pro Glu Pro Asn Leu Phe His Asp Gly Arg Glu His    2610 2615 2620 Ser Val His Val Glu Arg Thr Arg Gly Ile Phe Thr Val Gln Val Asp 2625 2630 2635 2640 Glu Asn Arg Arg Tyr Met Gln Asn Leu Thr Val Glu Gln Pro Ile Glu                2645 2650 2655 Val Lys Lys Leu Phe Val Gly Gly Ala Pro Pro Glu Phe Gln Pro Ser            2660 2665 2670 Pro Leu Arg Asn Ile Pro Pro Phe Glu Gly Cys Ile Trp Asn Leu Val        2675 2680 2685 Ile Asn Ser Val Pro Met Asp Phe Ala Arg Pro Val Ser Phe Lys Asn    2690 2695 2700 Ala Asp Ile Gly Arg Cys Ala His Gln Lys Leu Arg Glu Asp Glu Asp 2705 2710 2715 2720 Gly Ala Ala Pro Ala Glu Ile Val Ile Gln Pro Glu Pro Val Pro Thr                2725 2730 2735 Pro Ala Phe Pro Thr Pro Thr Pro Val Leu Thr His Gly Pro Cys Ala            2740 2745 2750 Ala Glu Ser Glu Pro Ala Leu Leu Ile Gly Ser Lys Gln Phe Gly Leu        2755 2760 2765 Ser Arg Asn Ser His Ile Ala Ile Ala Phe Asp Asp Thr Lys Val Lys    2770 2775 2780 Asn Arg Leu Thr Ile Glu Leu Glu Val Arg Thr Glu Ala Glu Ser Gly 2785 2790 2795 2800 Leu Leu Phe Tyr Met Ala Arg Ile Asn His Ala Asp Phe Ala Thr Val                2805 2810 2815 Gln Leu Arg Asn Gly Leu Pro Tyr Phe Ser Tyr Asp Leu Gly Ser Gly            2820 2825 2830 Asp Thr His Thr Met Ile Pro Thr Lys Ile Asn Asp Gly Gln Trp His        2835 2840 2845 Lys Ile Lys Ile Met Arg Ser Lys Gln Glu Gly Ile Leu Tyr Val Asp    2850 2855 2860 Gly Ala Ser Asn Arg Thr Ile Ser Pro Lys Lys Ala Asp Ile Leu Asp 2865 2870 2875 2880 Val Val Gly Met Leu Tyr Val Gly Gly Leu Pro Ile Asn Tyr Thr Thr                2885 2890 2895 Arg Arg Ile Gly Pro Val Thr Tyr Ser Ile Asp Gly Cys Val Arg Asn            2900 2905 2910 Leu His Met Ala Glu Ala Pro Ala Asp Leu Glu Gln Pro Thr Ser Ser        2915 2920 2925 Phe His Val Gly Thr Cys Phe Ala Asn Ala Gln Arg Gly Thr Tyr Phe    2930 2935 2940 Asp Gly Thr Gly Phe Ala Lys Ala Val Gly Gly Phe Lys Val Gly Leu 2945 2950 2955 2960 Asp Leu Leu Val Glu Phe Glu Phe Arg Thr Thr Thr Thr Thr Thr Gly Val                2965 2970 2975 Leu Leu Gly Ile Ser Ser Gln Lys Met Asp Gly Met Gly Ile Glu Met            2980 2985 2990 Ile Asp Glu Lys Leu Met Phe His Val Asp Asn Gly Ala Gly Arg Phe        2995 3000 3005 Thr Ala Val Tyr Asp Ala Gly Val Pro Gly His Leu Cys Asp Gly Gln    3010 3015 3020 Trp His Lys Val Thr Ala Asn Lys Ile Lys His Arg Ile Glu Leu Thr 3025 3030 3035 3040 Val Asp Gly Asn Gln Val Glu Ala Gln Ser Pro Asn Pro Ala Ser Thr                3045 3050 3055 Ser Ala Asp Thr Asn Asp Pro Val Phe Val Gly Gly Phe Pro Asp Asp            3060 3065 3070 Leu Lys Gln Phe Gly Leu Thr Thr Ser Ile Pro Phe Arg Gly Cys Ile        3075 3080 3085 Arg Ser Leu Lys Leu Thr Lys Gly Thr Gly Lys Pro Leu Glu Val Asn    3090 3095 3100 Phe Ala Lys Ala Leu Glu Leu Arg Gly Val Gln Pro Val Ser Cys Pro 3105 3110 3115 3120 Ala asn         <210> 11 <211> 3695 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit alpha-5 (Homo sapiens) <400> 11 Met Ala Lys Arg Leu Cys Ala Gly Ser Ala Leu Cys Val Arg Gly Pro   1 5 10 15 Arg Gly Pro Ala Pro Leu Leu Leu Val Gly Leu Ala Leu Leu Gly Ala              20 25 30 Ala Arg Ala Arg Glu Glu Ala Gly Gly Gly Phe Ser Leu His Pro Pro          35 40 45 Tyr Phe Asn Leu Ala Glu Gly Ala Arg Ile Ala Ala Ser Ala Thr Cys      50 55 60 Gly Glu Glu Ala Pro Ala Arg Gly Ser Pro Arg Pro Thr Glu Asp Leu  65 70 75 80 Tyr Cys Lys Leu Val Gly Gly Pro Val Ala Gly Gly Asp Pro Asn Gln                  85 90 95 Thr Ile Arg Gly Gln Tyr Cys Asp Ile Cys Thr Ala Ala Asn Ser Asn             100 105 110 Lys Ala His Pro Ala Ser Asn Ala Ile Asp Gly Thr Glu Arg Trp Trp         115 120 125 Gln Ser Pro Pro Leu Ser Arg Gly Leu Glu Tyr Asn Glu Val Asn Val     130 135 140 Thr Leu Asp Leu Gly Gln Val Phe His Val Ala Tyr Val Leu Ile Lys 145 150 155 160 Phe Ala Asn Ser Pro Arg Pro Asp Leu Trp Val Leu Glu Arg Ser Met                 165 170 175 Asp Phe Gly Arg Thr Tyr Gln Pro Trp Gln Phe Phe Ala Ser Ser Lys             180 185 190 Arg Asp Cys Leu Glu Arg Phe Gly Pro Gln Thr Leu Glu Arg Ile Thr         195 200 205 Arg Asp Asp Ala Ala Ile Cys Thr Thr Glu Tyr Ser Arg Ile Val Pro     210 215 220 Leu Glu Asn Gly Glu Ile Val Val Ser Leu Val Asn Gly Arg Pro Gly 225 230 235 240 Ala Met Asn Phe Ser Tyr Ser Pro Leu Leu Arg Glu Phe Thr Lys Ala                 245 250 255 Thr Asn Val Arg Leu Arg Phe Leu Arg Thr Asn Thr Leu Leu Gly His             260 265 270 Leu Met Gly Lys Ala Leu Arg Asp Pro Thr Val Thr Arg Arg Tyr Tyr         275 280 285 Tyr Ser Ile Lys Asp Ile Ser Ile Gly Gly Arg Cys Val Cys His Gly     290 295 300 His Ala Asp Ala Cys Asp Ala Lys Asp Pro Thr Asp Pro Phe Arg Leu 305 310 315 320 Gln Cys Thr Cys Gln His Asn Thr Cys Gly Gly Thr Cys Asp Arg Cys                 325 330 335 Cys Pro Gly Phe Asn Gln Gln Pro Trp Lys Pro Ala Thr Ala Asn Ser             340 345 350 Ala Asn Glu Cys Gln Ser Cys Asn Cys Tyr Gly His Ala Thr Asp Cys         355 360 365 Tyr Tyr Asp Pro Glu Val Asp Arg Arg Arg Ala Ser Gln Ser Leu Asp     370 375 380 Gly Thr Tyr Gln Gly Gly Gly Val Cys Ile Asp Cys Gln His His Thr 385 390 395 400 Thr Gly Val Asn Cys Glu Arg Cys Leu Pro Gly Phe Tyr Arg Ser Pro                 405 410 415 Asn His Pro Leu Asp Ser Pro His Val Cys Arg Arg Cys Asn Cys Glu             420 425 430 Ser Asp Phe Thr Asp Gly Thr Cys Glu Asp Leu Thr Gly Arg Cys Tyr         435 440 445 Cys Arg Pro Asn Phe Ser Gly Glu Arg Cys Asp Val Cys Ala Glu Gly     450 455 460 Phe Thr Gly Phe Pro Ser Cys Tyr Pro Thr Pro Ser Ser Ser Asn Asp 465 470 475 480 Thr Arg Glu Gln Val Leu Pro Ala Gly Gln Ile Val Asn Cys Asp Cys                 485 490 495 Ser Ala Ala Gly Thr Gln Gly Asn Ala Cys Arg Lys Asp Pro Arg Val             500 505 510 Gly Arg Cys Leu Cys Lys Pro Asn Phe Gln Gly Thr His Cys Glu Leu         515 520 525 Cys Ala Pro Gly Phe Tyr Gly Pro Gly Cys Gln Pro Cys Gln Cys Ser     530 535 540 Ser Pro Gly Val Ala Asp Asp Arg Cys Asp Pro Asp Thr Gly Gln Cys 545 550 555 560 Arg Cys Arg Val Gly Phe Glu Gly Ala Thr Cys Asp Arg Cys Ala Pro                 565 570 575 Gly Tyr Phe His Phe Pro Leu Cys Gln Leu Cys Gly Cys Ser Pro Ala             580 585 590 Gly Thr Leu Pro Glu Gly Cys Asp Glu Ala Gly Arg Cys Leu Cys Gln         595 600 605 Pro Glu Phe Ala Gly Pro His Cys Asp Arg Cys Arg Pro Gly Tyr His     610 615 620 Gly Phe Pro Asn Cys Gln Ala Cys Thr Cys Asp Pro Arg Gly Ala Leu 625 630 635 640 Asp Gln Leu Cys Gly Ala Gly Gly Leu Cys Arg Cys Arg Pro Gly Tyr                 645 650 655 Thr Gly Thr Ala Cys Gln Glu Cys Ser Pro Gly Phe His Gly Phe Pro             660 665 670 Ser Cys Val Pro Cys His Cys Ser Ala Glu Gly Ser Leu His Ala Ala         675 680 685 Cys Asp Pro Arg Ser Gly Gln Cys Ser Cys Arg Pro Arg Val Thr Gly     690 695 700 Leu Arg Cys Asp Thr Cys Val Pro Gly Ala Tyr Asn Phe Pro Tyr Cys 705 710 715 720 Glu Ala Gly Ser Cys His Pro Ala Gly Leu Ala Pro Val Asp Pro Ala                 725 730 735 Leu Pro Glu Ala Gln Val Pro Cys Met Cys Arg Ala His Val Glu Gly             740 745 750 Pro Ser Cys Asp Arg Cys Lys Pro Gly Phe Trp Gly Leu Ser Pro Ser         755 760 765 Asn Pro Glu Gly Cys Thr Arg Cys Ser Cys Asp Leu Arg Gly Thr Leu     770 775 780 Gly Gly Val Ala Glu Cys Gln Pro Gly Thr Gly Gln Cys Phe Cys Lys 785 790 795 800 Pro His Val Cys Gly Gln Ala Cys Ala Ser Cys Lys Asp Gly Phe Phe                 805 810 815 Gly Leu Asp Gln Ala Asp Tyr Phe Gly Cys Arg Ser Cys Arg Cys Asp             820 825 830 Ile Gly Gly Ala Leu Gly Gln Ser Cys Glu Pro Arg Thr Gly Val Cys         835 840 845 Arg Cys Arg Pro Asn Thr Gln Gly Pro Thr Cys Ser Glu Pro Ala Arg     850 855 860 Asp His Tyr Leu Pro Asp Leu His His Leu Arg Leu Glu Leu Glu Glu 865 870 875 880 Ala Ala Thr Pro Glu Gly His Ala Val Arg Phe Gly Phe Asn Pro Leu                 885 890 895 Glu Phe Glu Asn Phe Ser Trp Arg Gly Tyr Ala Gln Met Ala Pro Val             900 905 910 Gln Pro Arg Ile Val Ala Arg Leu Asn Leu Thr Ser Pro Asp Leu Phe         915 920 925 Trp Leu Val Phe Arg Tyr Val Asn Arg Gly Ala Met Ser Val Ser Gly     930 935 940 Arg Val Ser Val Arg Glu Glu Gly Arg Ser Ala Thr Cys Ala Asn Cys 945 950 955 960 Thr Ala Gln Ser Gln Pro Val Ala Phe Pro Pro Ser Thr Glu Pro Ala                 965 970 975 Phe Ile Thr Val Pro Gln Arg Gly Phe Gly Glu Pro Phe Val Leu Asn             980 985 990 Pro Gly Thr Trp Ala Leu Arg Val Glu Ala Glu Gly Val Leu Leu Asp         995 1000 1005 Tyr Val Val Leu Leu Pro Ser Ala Tyr Tyr Glu Ala Ala Leu Leu Gln    1010 1015 1020 Leu Arg Val Thr Glu Ala Cys Thr Tyr Arg Pro Ser Ala Gln Gln Ser 1025 1030 1035 1040 Gly Asp Asn Cys Leu Leu Tyr Thr His Leu Pro Leu Asp Gly Phe Pro                1045 1050 1055 Ser Ala Ala Gly Leu Glu Ala Leu Cys Arg Gln Asp Asn Ser Leu Pro            1060 1065 1070 Arg Pro Cys Pro Thr Glu Gln Leu Ser Pro Ser His Pro Pro Leu Ile        1075 1080 1085 Thr Cys Thr Gly Ser Asp Val Asp Val Gln Leu Gln Val Ala Val Pro    1090 1095 1100 Gln Pro Gly Arg Tyr Ala Leu Val Val Glu Tyr Ala Asn Glu Asp Ala 1105 1110 1115 1120 Arg Gln Glu Val Gly Val Ala Val His Thr Pro Gln Arg Ala Pro Gln                1125 1130 1135 Gln Gly Leu Leu Ser Leu His Pro Cys Leu Tyr Ser Thr Leu Cys Arg            1140 1145 1150 Gly Thr Ala Arg Asp Thr Gln Asp His Leu Ala Val Phe His Leu Asp        1155 1160 1165 Ser Glu Ala Ser Val Arg Leu Thr Ala Glu Gln Ala Arg Phe Phe Leu    1170 1175 1180 His Gly Val Thr Leu Val Pro Ile Glu Glu Phe Ser Pro Glu Phe Val 1185 1190 1195 1200 Glu Pro Arg Val Ser Cys Ile Ser Ser His Gly Ala Phe Gly Pro Asn                1205 1210 1215 Ser Ala Ala Cys Leu Pro Ser Arg Phe Pro Lys Pro Pro Gln Pro Ile            1220 1225 1230 Ile Leu Arg Asp Cys Gln Val Ile Pro Leu Pro Pro Gly Leu Pro Leu        1235 1240 1245 Thr His Ala Gln Asp Leu Thr Pro Ala Met Ser Pro Ala Gly Pro Arg    1250 1255 1260 Pro Arg Pro Pro Thr Ala Val Asp Pro Asp Ala Glu Pro Thr Leu Leu 1265 1270 1275 1280 Arg Glu Pro Gln Ala Thr Val Val Phe Thr Thr His Val Pro Thr Leu                1285 1290 1295 Gly Arg Tyr Ala Phe Leu Leu His Gly Tyr Gln Pro Ala His Pro Thr            1300 1305 1310 Phe Pro Val Glu Val Leu Ile Asn Ala Gly Arg Val Trp Gln Gly His        1315 1320 1325 Ala Asn Ala Ser Phe Cys Pro His Gly Tyr Gly Cys Arg Thr Leu Val    1330 1335 1340 Val Cys Glu Gly Gln Ala Leu Leu Asp Val Thr His Ser Glu Leu Thr 1345 1350 1355 1360 Val Thr Val Arg Val Pro Lys Gly Arg Trp Leu Trp Leu Asp Tyr Val                1365 1370 1375 Leu Val Val Pro Glu Asn Val Tyr Ser Phe Gly Tyr Leu Arg Glu Glu            1380 1385 1390 Pro Leu Asp Lys Ser Tyr Asp Phe Ile Ser His Cys Ala Ala Gln Gly        1395 1400 1405 Tyr His Ile Ser Pro Ser Ser Ser Ser Leu Phe Cys Arg Asn Ala Ala    1410 1415 1420 Ala Ser Leu Ser Leu Phe Tyr Asn Asn Gly Ala Arg Pro Cys Gly Cys 1425 1430 1435 1440 His Glu Val Gly Ala Thr Gly Pro Thr Cys Glu Pro Phe Gly Gly Gln                1445 1450 1455 Cys Pro Cys His Ala His Val Ile Gly Arg Asp Cys Ser Arg Cys Ala            1460 1465 1470 Thr Gly Tyr Trp Gly Phe Pro Asn Cys Arg Pro Cys Asp Cys Gly Ala        1475 1480 1485 Arg Leu Cys Asp Glu Leu Thr Gly Gln Cys Ile Cys Pro Pro Arg Thr    1490 1495 1500 Ile Pro Pro Asp Cys Leu Leu Cys Gln Pro Gln Thr Phe Gly Cys His 1505 1510 1515 1520 Pro Leu Val Gly Cys Glu Glu Cys Asn Cys Ser Gly Pro Gly Ile Gln                1525 1530 1535 Glu Leu Thr Asp Pro Thr Cys Asp Thr Asp Ser Gly Gln Cys Lys Cys            1540 1545 1550 Arg Pro Asn Val Thr Gly Arg Arg Cys Asp Thr Cys Ser Pro Gly Phe        1555 1560 1565 His Gly Tyr Pro Arg Cys Arg Pro Cys Asp Cys His Glu Ala Gly Thr    1570 1575 1580 Ala Pro Gly Val Cys Asp Pro Leu Thr Gly Gln Cys Tyr Cys Lys Glu 1585 1590 1595 1600 Asn Val Gln Gly Pro Lys Cys Asp Gln Cys Ser Leu Gly Thr Phe Ser                1605 1610 1615 Leu Asp Ala Ala Asn Pro Lys Gly Cys Thr Arg Cys Phe Cys Phe Gly            1620 1625 1630 Ala Thr Glu Arg Cys Arg Ser Ser Ser Tyr Thr Arg Gln Glu Phe Val        1635 1640 1645 Asp Met Glu Gly Trp Val Leu Leu Ser Thr Asp Arg Gln Val Val Pro    1650 1655 1660 His Glu Arg Gln Pro Gly Thr Glu Met Leu Arg Ala Asp Leu Arg His 1665 1670 1675 1680 Val Pro Glu Ala Val Pro Glu Ala Phe Pro Glu Leu Tyr Trp Gln Ala                1685 1690 1695 Pro Pro Ser Tyr Leu Gly Asp Arg Val Ser Ser Tyr Gly Gly Thr Leu            1700 1705 1710 Arg Tyr Glu Leu His Ser Glu Thr Gln Arg Gly Asp Val Phe Val Pro        1715 1720 1725 Met Glu Ser Arg Pro Asp Val Val Leu Gln Gly Asn Gln Met Ser Ile    1730 1735 1740 Thr Phe Leu Glu Pro Ala Tyr Pro Thr Pro Gly His Val His Arg Gly 1745 1750 1755 1760 Gln Leu Gln Leu Val Glu Gly Asn Phe Arg His Thr Glu Thr Arg Asn                1765 1770 1775 Thr Val Ser Arg Glu Glu Leu Met Met Val Leu Ala Ser Leu Glu Gln            1780 1785 1790 Leu Gln Ile Arg Ala Leu Phe Ser Gln Ile Ser Ser Ala Val Phe Leu        1795 1800 1805 Arg Arg Val Ala Leu Glu Val Ala Ser Pro Ala Gly Gln Gly Ala Leu    1810 1815 1820 Ala Ser Asn Val Glu Leu Cys Leu Cys Pro Ala Ser Tyr Arg Gly Asp 1825 1830 1835 1840 Ser Cys Gln Glu Cys Ala Pro Gly Phe Tyr Arg Asp Val Lys Gly Leu                1845 1850 1855 Phe Leu Gly Arg Cys Val Pro Cys Gln Cys His Gly His Ser Asp Arg            1860 1865 1870 Cys Leu Pro Gly Ser Gly Val Cys Val Asp Cys Gln His Asn Thr Glu        1875 1880 1885 Gly Ala His Cys Glu Arg Cys Gln Ala Gly Phe Val Ser Ser Arg Asp    1890 1895 1900 Asp Pro Ser Ala Pro Cys Val Ser Cys Pro Cys Pro Leu Ser Val Pro 1905 1910 1915 1920 Ser Asn Asn Phe Ala Glu Gly Cys Val Leu Arg Gly Gly Arg Thr Gln                1925 1930 1935 Cys Leu Cys Lys Pro Gly Tyr Ala Gly Ala Ser Cys Glu Arg Cys Ala            1940 1945 1950 Pro Gly Phe Phe Gly Asn Pro Leu Val Leu Gly Ser Ser Cys Gln Pro        1955 1960 1965 Cys Asp Cys Ser Gly Asn Gly Asp Pro Asn Leu Leu Phe Ser Asp Cys    1970 1975 1980 Asp Pro Leu Thr Gly Ala Cys Arg Gly Cys Leu Arg His Thr Thr Gly 1985 1990 1995 2000 Pro Arg Cys Glu Ile Cys Ala Pro Gly Phe Tyr Gly Asn Ala Leu Leu                2005 2010 2015 Pro Gly Asn Cys Thr Arg Cys Asp Cys Thr Pro Cys Gly Thr Glu Ala            2020 2025 2030 Cys Asp Pro His Ser Gly His Cys Leu Cys Lys Ala Gly Val Thr Gly        2035 2040 2045 Arg Arg Cys Asp Arg Cys Gln Glu Gly His Phe Gly Phe Asp Gly Cys    2050 2055 2060 Gly Gly Cys Arg Pro Cys Ala Cys Gly Pro Ala Ala Glu Gly Ser Glu 2065 2070 2075 2080 Cys His Pro Gln Ser Gly Gln Cys His Cys Arg Pro Gly Thr Met Gly                2085 2090 2095 Pro Gln Cys Arg Glu Cys Ala Pro Gly Tyr Trp Gly Leu Pro Glu Gln            2100 2105 2110 Gly Cys Arg Arg Cys Gln Cys Pro Gly Gly Arg Cys Asp Pro His Thr        2115 2120 2125 Gly Arg Cys Asn Cys Pro Pro Gly Leu Ser Gly Glu Arg Cys Asp Thr    2130 2135 2140 Cys Ser Gln Gln His Gln Val Pro Val Pro Gly Gly Pro Val Gly His 2145 2150 2155 2160 Ser Ile His Cys Glu Val Cys Asp His Cys Val Val Leu Leu Leu Asp                2165 2170 2175 Asp Leu Glu Arg Ala Gly Ala Leu Leu Pro Ala Ile His Glu Gln Leu            2180 2185 2190 Arg Gly Ile Asn Ala Ser Ser Met Ala Trp Ala Arg Leu His Arg Leu        2195 2200 2205 Asn Ala Ser Ile Ala Asp Leu Gln Ser Gln Leu Arg Ser Pro Leu Gly    2210 2215 2220 Pro Arg His Glu Thr Ala Gln Gln Leu Glu Val Leu Glu Gln Gln Ser 2225 2230 2235 2240 Thr Ser Leu Gly Gln Asp Ala Arg Arg Leu Gly Gly Gln Ala Val Gly                2245 2250 2255 Thr Arg Asp Gln Ala Ser Gln Leu Leu Ala Gly Thr Glu Ala Thr Leu            2260 2265 2270 Gly His Ala Lys Thr Leu Leu Ala Ala Ile Arg Ala Val Asp Arg Thr        2275 2280 2285 Leu Ser Glu Leu Met Ser Gln Thr Gly His Leu Gly Leu Ala Asn Ala    2290 2295 2300 Ser Ala Pro Ser Gly Glu Gln Leu Leu Arg Thr Leu Ala Glu Val Glu 2305 2310 2315 2320 Arg Leu Leu Trp Glu Met Arg Ala Arg Asp Leu Gly Ala Pro Gln Ala                2325 2330 2335 Ala Ala Glu Ala Glu Leu Ala Ala Ala Gln Arg Leu Leu Ala Arg Val            2340 2345 2350 Gln Glu Gln Leu Ser Ser Leu Trp Glu Glu Asn Gln Ala Leu Ala Thr        2355 2360 2365 Gln Thr Arg Asp Arg Leu Ala Gln His Glu Ala Gly Leu Met Asp Leu    2370 2375 2380 Arg Glu Ala Leu Asn Arg Ala Val Asp Ala Thr Arg Glu Ala Gln Glu 2385 2390 2395 2400 Leu Asn Ser Arg Asn Gln Glu Arg Leu Glu Glu Ala Leu Gln Arg Lys                2405 2410 2415 Gln Glu Leu Ser Arg Asp Asn Ala Thr Leu Gln Ala Thr Leu His Ala            2420 2425 2430 Ala Arg Asp Thr Leu Ala Ser Val Phe Arg Leu Leu His Ser Leu Asp        2435 2440 2445 Gln Ala Lys Glu Glu Leu Glu Arg Leu Ala Ala Ser Leu Asp Gly Ala    2450 2455 2460 Arg Thr Pro Leu Leu Gln Arg Met Gln Thr Phe Ser Pro Ala Gly Ser 2465 2470 2475 2480 Lys Leu Arg Leu Val Glu Ala Ala Glu Ala His Ala Gln Gln Leu Gly                2485 2490 2495 Gln Leu Ala Leu Asn Leu Ser Ser Ile Ile Leu Asp Val Asn Gln Asp            2500 2505 2510 Arg Leu Thr Gln Arg Ala Ile Glu Ala Ser Asn Ala Tyr Ser Arg Ile        2515 2520 2525 Leu Gln Ala Val Gln Ala Ala Glu Asp Ala Ala Gly Gln Ala Leu Gln    2530 2535 2540 Gln Ala Asp His Thr Trp Ala Thr Val Val Arg Gln Gly Leu Val Asp 2545 2550 2555 2560 Arg Ala Gln Gln Leu Leu Ala Asn Ser Thr Ala Leu Glu Glu Ala Met                2565 2570 2575 Leu Gln Glu Gln Gln Arg Leu Gly Leu Val Trp Ala Ala Leu Gln Gly            2580 2585 2590 Ala Arg Thr Gln Leu Arg Asp Val Arg Ala Lys Lys Asp Gln Leu Glu        2595 2600 2605 Ala His Ile Gln Ala Ala Gln Ala Met Leu Ala Met Asp Thr Asp Glu    2610 2615 2620 Thr Ser Lys Lys Ile Ala His Ala Lys Ala Val Ala Ala Glu Ala Gln 2625 2630 2635 2640 Asp Thr Ala Thr Arg Val Gln Ser Gln Leu Gln Ala Met Gln Glu Asn                2645 2650 2655 Val Glu Arg Trp Gln Gly Gln Tyr Glu Gly Leu Arg Gly Gln Asp Leu            2660 2665 2670 Gly Gln Ala Val Leu Asp Ala Gly His Ser Val Ser Thr Leu Glu Lys        2675 2680 2685 Thr Leu Pro Gln Leu Leu Ala Lys Leu Ser Ile Leu Glu Asn Arg Gly    2690 2695 2700 Val His Asn Ala Ser Leu Ala Leu Ser Ala Ser Ile Gly Arg Val Arg 2705 2710 2715 2720 Glu Leu Ile Ala Gln Ala Arg Gly Ala Ala Ser Lys Val Lys Val Pro                2725 2730 2735 Met Lys Phe Asn Gly Arg Ser Gly Val Gln Leu Arg Thr Pro Arg Asp            2740 2745 2750 Leu Ala Asp Leu Ala Ala Tyr Thr Ala Leu Lys Phe Tyr Leu Gln Gly        2755 2760 2765 Pro Glu Pro Glu Pro Gly Gln Gly Thr Glu Asp Arg Phe Val Met Tyr    2770 2775 2780 Met Gly Ser Arg Gln Ala Thr Gly Asp Tyr Met Gly Val Ser Leu Arg 2785 2790 2795 2800 Asp Lys Lys Val His Trp Val Tyr Gln Leu Gly Glu Ala Gly Pro Ala                2805 2810 2815 Val Leu Ser Ile Asp Glu Asp Ile Gly Glu Gln Phe Ala Ala Val Ser            2820 2825 2830 Leu Asp Arg Thr Leu Gln Phe Gly His Met Ser Val Thr Val Glu Arg        2835 2840 2845 Gln Met Ile Gln Glu Thr Lys Gly Asp Thr Val Ala Pro Gly Ala Glu    2850 2855 2860 Gly Leu Leu Asn Leu Arg Pro Asp Asp Phe Val Phe Tyr Val Gly Gly 2865 2870 2875 2880 Tyr Pro Ser Thr Phe Thr Pro Pro Pro Leu Leu Arg Phe Pro Gly Tyr                2885 2890 2895 Arg Gly Cys Ile Glu Met Asp Thr Leu Asn Glu Glu Val Val Ser Leu            2900 2905 2910 Tyr Asn Phe Glu Arg Thr Phe Gln Leu Asp Thr Ala Val Asp Arg Pro        2915 2920 2925 Cys Ala Arg Ser Lys Ser Thr Gly Asp Pro Trp Leu Thr Asp Gly Ser    2930 2935 2940 Tyr Leu Asp Gly Thr Gly Phe Ala Arg Ile Ser Phe Asp Ser Gln Ile 2945 2950 2955 2960 Ser Thr Thr Lys Arg Phe Glu Gln Glu Leu Arg Leu Val Ser Tyr Ser                2965 2970 2975 Gly Val Leu Phe Phe Leu Lys Gln Gln Ser Gln Phe Leu Cys Leu Ala            2980 2985 2990 Val Gln Glu Gly Ser Leu Val Leu Leu Tyr Asp Phe Gly Ala Gly Leu        2995 3000 3005 Lys Lys Ala Val Pro Leu Gln Pro Pro Pro Pro Leu Thr Ser Ala Ser    3010 3015 3020 Lys Ala Ile Gln Val Phe Leu Leu Gly Gly Ser Arg Lys Arg Val Leu 3025 3030 3035 3040 Val Arg Val Glu Arg Ala Thr Val Tyr Ser Val Glu Gln Asp Asn Asp                3045 3050 3055 Leu Glu Leu Ala Asp Ala Tyr Tyr Leu Gly Gly Val Pro Pro Asp Gln            3060 3065 3070 Leu Pro Pro Ser Leu Arg Arg Leu Phe Pro Thr Gly Gly Ser Val Arg        3075 3080 3085 Gly Cys Val Lys Gly Ile Lys Ala Leu Gly Lys Tyr Val Asp Leu Lys    3090 3095 3100 Arg Leu Asn Thr Thr Gly Val Ser Ala Gly Cys Thr Ala Asp Leu Leu 3105 3110 3115 3120 Val Gly Arg Ala Met Thr Phe His Gly His Gly Phe Leu Arg Leu Ala                3125 3130 3135 Leu Ser Asn Val Ala Pro Leu Thr Gly Asn Val Tyr Ser Gly Phe Gly            3140 3145 3150 Phe His Ser Ala Gln Asp Ser Ala Leu Leu Tyr Tyr Arg Ala Ser Pro        3155 3160 3165 Asp Gly Leu Cys Gln Val Ser Leu Gln Gln Gly Arg Val Ser Leu Gln    3170 3175 3180 Leu Leu Arg Thr Glu Val Lys Thr Gln Ala Gly Phe Ala Asp Gly Ala 3185 3190 3195 3200 Pro His Tyr Val Ala Phe Tyr Ser Asn Ala Thr Gly Val Trp Leu Tyr                3205 3210 3215 Val Asp Asp Gln Leu Gln Gln Met Lys Pro His Arg Gly Pro Pro Pro            3220 3225 3230 Glu Leu Gln Pro Gln Pro Glu Gly Pro Pro Arg Leu Leu Leu Gly Gly        3235 3240 3245 Leu Pro Glu Ser Gly Thr Ile Tyr Asn Phe Ser Gly Cys Ile Ser Asn    3250 3255 3260 Val Phe Val Gln Arg Leu Leu Gly Pro Gln Arg Val Phe Asp Leu Gln 3265 3270 3275 3280 Gln Asn Leu Gly Ser Val Asn Val Ser Thr Gly Cys Ala Pro Ala Leu                3285 3290 3295 Gln Ala Gln Thr Pro Gly Leu Gly Pro Arg Gly Leu Gln Ala Thr Ala            3300 3305 3310 Arg Lys Ala Ser Arg Arg Ser Arg Gln Pro Ala Arg His Pro Ala Cys        3315 3320 3325 Met Leu Pro Pro His Leu Arg Thr Thr Arg Asp Ser Tyr Gln Phe Gly    3330 3335 3340 Gly Ser Leu Ser Ser His Leu Glu Phe Val Gly Ile Leu Ala Arg His 3345 3350 3355 3360 Arg Asn Trp Pro Ser Leu Ser Met His Val Leu Pro Arg Ser Ser Arg                3365 3370 3375 Gly Leu Leu Leu Phe Thr Ala Arg Leu Arg Pro Gly Ser Pro Ser Leu            3380 3385 3390 Ala Leu Phe Leu Ser Asn Gly His Phe Val Ala Gln Met Glu Gly Leu        3395 3400 3405 Gly Thr Arg Leu Arg Ala Gln Ser Arg Gln Arg Ser Arg Pro Gly Arg    3410 3415 3420 Trp His Lys Val Ser Val Arg Trp Glu Lys Asn Arg Ile Leu Leu Val 3425 3430 3435 3440 Thr Asp Gly Ala Arg Ala Trp Ser Gln Glu Gly Pro His Arg Gln His                3445 3450 3455 Gln Gly Ala Glu His Pro Gln Pro His Thr Leu Phe Val Gly Gly Leu            3460 3465 3470 Pro Ala Ser Ser His Ser Ser Lys Leu Pro Val Thr Val Gly Phe Ser        3475 3480 3485 Gly Cys Val Lys Arg Leu Arg Leu His Gly Arg Pro Leu Gly Ala Pro    3490 3495 3500 Thr Arg Met Ala Gly Val Thr Pro Cys Ile Leu Gly Pro Leu Glu Ala 3505 3510 3515 3520 Gly Leu Phe Phe Pro Gly Ser Gly Gly Val Ile Thr Leu Asp Leu Pro                3525 3530 3535 Gly Ala Thr Leu Pro Asp Val Gly Leu Glu Leu Glu Val Arg Pro Leu            3540 3545 3550 Ala Val Thr Gly Leu Ile Phe His Leu Gly Gln Ala Arg Thr Pro Pro        3555 3560 3565 Tyr Leu Gln Leu Gln Val Thr Glu Lys Gln Val Leu Leu Arg Ala Asp    3570 3575 3580 Asp Gly Ala Gly Glu Phe Ser Thr Ser Val Thr Arg Pro Ser Val Leu 3585 3590 3595 3600 Cys Asp Gly Gln Trp His Arg Leu Ala Val Met Lys Ser Gly Asn Val                3605 3610 3615 Leu Arg Leu Glu Val Asp Ala Gln Ser Asn His Thr Val Gly Pro Leu            3620 3625 3630 Leu Ala Ala Ala Ala Gly Ala Pro Ala Pro Leu Tyr Leu Gly Gly Leu        3635 3640 3645 Pro Glu Pro Met Ala Val Gln Pro Trp Pro Pro Ala Tyr Cys Gly Cys    3650 3655 3660 Met Arg Arg Leu Ala Val Asn Arg Ser Pro Val Ala Met Thr Arg Ser 3665 3670 3675 3680 Val Glu Val His Gly Ala Val Gly Ala Ser Gly Cys Pro Ala Ala                3685 3690 3695 <210> 12 <211> 1786 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Laminin subunit beta-1 (Homo sapiens) <400> 12 Met Gly Leu Leu Gln Leu Leu Ala Phe Ser Phe Leu Ala Leu Cys Arg   1 5 10 15 Ala Arg Val Arg Ala Gln Glu Pro Glu Phe Ser Tyr Gly Cys Ala Glu              20 25 30 Gly Ser Cys Tyr Pro Ala Thr Gly Asp Leu Leu Ile Gly Arg Ala Gln          35 40 45 Lys Leu Ser Val Thr Ser Thr Cys Gly Leu His Lys Pro Glu Pro Tyr      50 55 60 Cys Ile Val Ser His Leu Gln Glu Asp Lys Lys Cys Phe Ile Cys Asn  65 70 75 80 Ser Gln Asp Pro Tyr His Glu Thr Leu Asn Pro Asp Ser His Leu Ile                  85 90 95 Glu Asn Val Val Thr Thr Phe Ala Pro Asn Arg Leu Lys Ile Trp Trp             100 105 110 Gln Ser Glu Asn Gly Val Glu Asn Val Thr Ile Gln Leu Asp Leu Glu         115 120 125 Ala Glu Phe His Phe Thr His Leu Ile Met Thr Phe Lys Thr Phe Arg     130 135 140 Pro Ala Ala Met Leu Ile Glu Arg Ser Ser Asp Phe Gly Lys Thr Trp 145 150 155 160 Gly Val Tyr Arg Tyr Phe Ala Tyr Asp Cys Glu Ala Ser Phe Pro Gly                 165 170 175 Ile Ser Thr Gly Pro Met Lys Lys Val Asp Asp Ile Ile Cys Asp Ser             180 185 190 Arg Tyr Ser Asp Ile Glu Pro Ser Thr Glu Gly Glu Val Ile Phe Arg         195 200 205 Ala Leu Asp Pro Ala Phe Lys Ile Glu Asp Pro Tyr Ser Pro Arg Ile     210 215 220 Gln Asn Leu Leu Lys Ile Thr Asn Leu Arg Ile Lys Phe Val Lys Leu 225 230 235 240 His Thr Leu Gly Asp Asn Leu Leu Asp Ser Arg Met Glu Ile Arg Glu                 245 250 255 Lys Tyr Tyr Tyr Ala Val Tyr Asp Met Val Val Arg Gly Asn Cys Phe             260 265 270 Cys Tyr Gly His Ala Ser Glu Cys Ala Pro Val Asp Gly Phe Asn Glu         275 280 285 Glu Val Glu Gly Met Val His Gly His Cys Met Cys Arg His Asn Thr     290 295 300 Lys Gly Leu Asn Cys Glu Leu Cys Met Asp Phe Tyr His Asp Leu Pro 305 310 315 320 Trp Arg Pro Ala Glu Gly Arg Asn Ser Asn Ala Cys Lys Lys Cys Asn                 325 330 335 Cys Asn Glu His Ser Ile Ser Cys His Phe Asp Met Ala Val Tyr Leu             340 345 350 Ala Thr Gly Asn Val Ser Gly Gly Val Cys Asp Asp Cys Gln His Asn         355 360 365 Thr Met Gly Arg Asn Cys Glu Gln Cys Lys Pro Phe Tyr Tyr Gln His     370 375 380 Pro Glu Arg Asp Ile Arg Asp Pro Asn Phe Cys Glu Arg Cys Thr Cys 385 390 395 400 Asp Pro Ala Gly Ser Gln Asn Glu Gly Ile Cys Asp Ser Tyr Thr Asp                 405 410 415 Phe Ser Thr Gly Leu Ile Ala Gly Gln Cys Arg Cys Lys Leu Asn Val             420 425 430 Glu Gly Glu His Cys Asp Val Cys Lys Glu Gly Phe Tyr Asp Leu Ser         435 440 445 Ser Glu Asp Pro Phe Gly Cys Lys Ser Cys Ala Cys Asn Pro Leu Gly     450 455 460 Thr Ile Pro Gly Gly Asn Pro Cys Asp Ser Glu Thr Gly His Cys Tyr 465 470 475 480 Cys Lys Arg Leu Val Thr Gly Gln His Cys Asp Gln Cys Leu Pro Glu                 485 490 495 His Trp Gly Leu Ser Asn Asp Leu Asp Gly Cys Arg Pro Cys Asp Cys             500 505 510 Asp Leu Gly Gly Ala Leu Asn Asn Ser Cys Phe Ala Glu Ser Gly Gln         515 520 525 Cys Ser Cys Arg Pro His Met Ile Gly Arg Gln Cys Asn Glu Val Glu     530 535 540 Pro Gly Tyr Tyr Phe Ala Thr Leu Asp His Tyr Leu Tyr Glu Ala Glu 545 550 555 560 Glu Ala Asn Leu Gly Pro Gly Val Ser Ile Val Glu Arg Gln Tyr Ile                 565 570 575 Gln Asp Arg Ile Pro Ser Trp Thr Gly Ala Gly Phe Val Arg Val Pro             580 585 590 Glu Gly Ala Tyr Leu Glu Phe Phe Ile Asp Asn Ile Pro Tyr Ser Met         595 600 605 Glu Tyr Asp Ile Leu Ile Arg Tyr Glu Pro Gln Leu Pro Asp His Trp     610 615 620 Glu Lys Ala Val Ile Thr Val Gln Arg Pro Gly Arg Ile Pro Thr Ser 625 630 635 640 Ser Arg Cys Gly Asn Thr Ile Pro Asp Asp Asp Asn Gln Val Val Ser                 645 650 655 Leu Ser Pro Gly Ser Arg Tyr Val Val Leu Pro Arg Pro Val Cys Phe             660 665 670 Glu Lys Gly Thr Asn Tyr Thr Val Arg Leu Glu Leu Pro Gln Tyr Thr         675 680 685 Ser Ser Asp Ser Asp Val Glu Ser Pro Tyr Thr Leu Ile Asp Ser Leu     690 695 700 Val Leu Met Pro Tyr Cys Lys Ser Leu Asp Ile Phe Thr Val Gly Gly 705 710 715 720 Ser Gly Asp Gly Val Val Thr Asn Ser Ala Trp Glu Thr Phe Gln Arg                 725 730 735 Tyr Arg Cys Leu Glu Asn Ser Arg Ser Val Val Lys Thr Pro Met Thr             740 745 750 Asp Val Cys Arg Asn Ile Ile Phe Ser Ile Ser Ala Leu Leu His Gln         755 760 765 Thr Gly Leu Ala Cys Glu Cys Asp Pro Gln Gly Ser Leu Ser Ser Val     770 775 780 Cys Asp Pro Asn Gly Gly Gln Cys Gln Cys Arg Pro Asn Val Val Gly 785 790 795 800 Arg Thr Cys Asn Arg Cys Ala Pro Gly Thr Phe Gly Phe Gly Pro Ser                 805 810 815 Gly Cys Lys Pro Cys Glu Cys His Leu Gln Gly Ser Val Asn Ala Phe             820 825 830 Cys Asn Pro Val Thr Gly Gln Cys His Cys Phe Gln Gly Val Tyr Ala         835 840 845 Arg Gln Cys Asp Arg Cys Leu Pro Gly His Trp Gly Phe Pro Ser Cys     850 855 860 Gln Pro Cys Gln Cys Asn Gly His Ala Asp Asp Cys Asp Pro Val Thr 865 870 875 880 Gly Glu Cys Leu Asn Cys Gln Asp Tyr Thr Met Gly His Asn Cys Glu                 885 890 895 Arg Cys Leu Ala Gly Tyr Tyr Gly Asp Pro Ile Ile Gly Ser Gly Asp             900 905 910 His Cys Arg Pro Cys Pro Cys Pro Asp Gly Pro Asp Ser Gly Arg Gln         915 920 925 Phe Ala Arg Ser Cys Tyr Gln Asp Pro Val Thr Leu Gln Leu Ala Cys     930 935 940 Val Cys Asp Pro Gly Tyr Ile Gly Ser Arg Cys Asp Asp Cys Ala Ser 945 950 955 960 Gly Tyr Phe Gly Asn Pro Ser Glu Val Gly Gly Ser Cys Gln Pro Cys                 965 970 975 Gln Cys His Asn Asn Ile Asp Thr Thr Asp Pro Glu Ala Cys Asp Lys             980 985 990 Glu Thr Gly Arg Cys Leu Lys Cys Leu Tyr His Thr Glu Gly Glu His         995 1000 1005 Cys Gln Phe Cys Arg Phe Gly Tyr Tyr Gly Asp Ala Leu Gln Gln Asp    1010 1015 1020 Cys Arg Lys Cys Val Cys Asn Tyr Leu Gly Thr Val Gln Glu His Cys 1025 1030 1035 1040 Asn Gly Ser Asp Cys Gln Cys Asp Lys Ala Thr Gly Gln Cys Leu Cys                1045 1050 1055 Leu Pro Asn Val Ile Gly Gln Asn Cys Asp Arg Cys Ala Pro Asn Thr            1060 1065 1070 Trp Gln Leu Ala Ser Gly Thr Gly Cys Asp Pro Cys Asn Cys Asn Ala        1075 1080 1085 Ala His Ser Phe Gly Pro Ser Cys Asn Glu Phe Thr Gly Gln Cys Gln    1090 1095 1100 Cys Met Pro Gly Phe Gly Gly Arg Thr Cys Ser Glu Cys Gln Glu Leu 1105 1110 1115 1120 Phe Trp Gly Asp Pro Asp Val Glu Cys Arg Ala Cys Asp Cys Asp Pro                1125 1130 1135 Arg Gly Ile Glu Thr Pro Gln Cys Asp Gln Ser Thr Gly Gln Cys Val            1140 1145 1150 Cys Val Glu Gly Val Glu Gly Pro Arg Cys Asp Lys Cys Thr Arg Gly        1155 1160 1165 Tyr Ser Gly Val Phe Pro Asp Cys Thr Pro Cys His Gln Cys Phe Ala    1170 1175 1180 Leu Trp Asp Val Ile Ile Ala Glu Leu Thr Asn Arg Thr His Arg Phe 1185 1190 1195 1200 Leu Glu Lys Ala Lys Ala Leu Lys Ile Ser Gly Val Ile Gly Pro Tyr                1205 1210 1215 Arg Glu Thr Val Asp Ser Val Glu Arg Lys Val Ser Glu Ile Lys Asp            1220 1225 1230 Ile Leu Ala Gln Ser Pro Ala Ala Glu Pro Leu Lys Asn Ile Gly Asn        1235 1240 1245 Leu Phe Glu Glu Ala Glu Lys Leu Ile Lys Asp Val Thr Glu Met Met    1250 1255 1260 Ala Gln Val Glu Val Lys Leu Ser Asp Thr Thr Ser Gln Ser Asn Ser 1265 1270 1275 1280 Thr Ala Lys Glu Leu Asp Ser Leu Gln Thr Glu Ala Glu Ser Leu Asp                1285 1290 1295 Asn Thr Val Lys Glu Leu Ala Glu Gln Leu Glu Phe Ile Lys Asn Ser            1300 1305 1310 Asp Ile Arg Gly Ala Leu Asp Ser Ile Thr Lys Tyr Phe Gln Met Ser        1315 1320 1325 Leu Glu Ala Glu Glu Arg Val Asn Ala Ser Thr Thr Glu Pro Asn Ser    1330 1335 1340 Thr Val Glu Gln Ser Ala Leu Met Arg Asp Arg Val Glu Asp Val Met 1345 1350 1355 1360 Met Glu Arg Glu Ser Gln Phe Lys Glu Lys Gln Glu Glu Gln Ala Arg                1365 1370 1375 Leu Leu Asp Glu Leu Ala Gly Lys Leu Gln Ser Leu Asp Leu Ser Ala            1380 1385 1390 Ala Ala Glu Met Thr Cys Gly Thr Pro Pro Gly Ala Ser Cys Ser Glu        1395 1400 1405 Thr Glu Cys Gly Gly Pro Asn Cys Arg Thr Asp Glu Gly Glu Arg Lys    1410 1415 1420 Cys Gly Gly Pro Gly Cys Gly Gly Leu Val Thr Val Ala His Asn Ala 1425 1430 1435 1440 Trp Gln Lys Ala Met Asp Leu Asp Gln Asp Val Leu Ser Ala Leu Ala                1445 1450 1455 Glu Val Glu Gln Leu Ser Lys Met Val Ser Glu Ala Lys Leu Arg Ala            1460 1465 1470 Asp Glu Ala Lys Gln Ser Ala Glu Asp Ile Leu Leu Lys Thr Asn Ala        1475 1480 1485 Thr Lys Glu Lys Met Asp Lys Ser Asn Glu Glu Leu Arg Asn Leu Ile    1490 1495 1500 Lys Gln Ile Arg Asn Phe Leu Thr Gln Asp Ser Ala Asp Leu Asp Ser 1505 1510 1515 1520 Ile Glu Ala Val Ala Asn Glu Val Leu Lys Met Glu Met Pro Ser Thr                1525 1530 1535 Pro Gln Gln Leu Gln Asn Leu Thr Glu Asp Ile Arg Glu Arg Val Glu            1540 1545 1550 Ser Leu Ser Gln Val Glu Val Ile Leu Gln His Ser Ala Ala Asp Ile        1555 1560 1565 Ala Arg Ala Glu Met Leu Leu Glu Glu Ala Lys Arg Ala Ser Lys Ser    1570 1575 1580 Ala Thr Asp Val Lys Val Thr Ala Asp Met Val Lys Glu Ala Leu Glu 1585 1590 1595 1600 Glu Ala Glu Lys Ala Gln Val Ala Ala Glu Lys Ala Ile Lys Gln Ala                1605 1610 1615 Asp Glu Asp Ile Gln Gly Thr Gln Asn Leu Leu Thr Ser Ile Glu Ser            1620 1625 1630 Glu Thr Ala Ala Ser Glu Glu Thr Leu Phe Asn Ala Ser Gln Arg Ile        1635 1640 1645 Ser Glu Leu Glu Arg Asn Val Glu Glu Leu Lys Arg Lys Ala Ala Gln    1650 1655 1660 Asn Ser Gly Glu Ala Glu Tyr Ile Glu Lys Val Val Tyr Thr Val Lys 1665 1670 1675 1680 Gln Ser Ala Glu Asp Val Lys Lys Thr Leu Asp Gly Glu Leu Asp Glu                1685 1690 1695 Lys Tyr Lys Lys Val Glu Asn Leu Ile Ala Lys Lys Thr Glu Glu Ser            1700 1705 1710 Ala Asp Ala Arg Arg Lys Ala Glu Met Leu Gln Asn Glu Ala Lys Thr        1715 1720 1725 Leu Leu Ala Gln Ala Asn Ser Lys Leu Gln Leu Leu Lys Asp Leu Glu    1730 1735 1740 Arg Lys Tyr Glu Asp Asn Gln Arg Tyr Leu Glu Asp Lys Ala Gln Glu 1745 1750 1755 1760 Leu Ala Arg Leu Glu Gly Glu Val Arg Ser Leu Leu Lys Asp Ile Ser                1765 1770 1775 Gln Lys Val Ala Val Tyr Ser Thr Cys Leu            1780 1785 <210> 13 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> siRNA sequence of KRS <400> 13 agaaguuuuc aucuaugaac auggc 25 <210> 14 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> siRNA sequence of LRS <400> 14 ccuugcaugg aucaugauag acaaa 25

Claims (10)

A pharmaceutical composition for preventing or treating immune cell migration-related diseases comprising a compound of Formula 1-1 or a pharmaceutically acceptable salt thereof as an active ingredient,

<Formula 1-1>

In Chemical Formula 1-1,
R 1 is hydrogen; Or a halogen group,
R 4 is hydrogen; Or a C1-C6 alkoxy group
R7 is hydrogen; Or a hydroxycarbonyl group,

The immune cell migration-related disease
(i) cardiovascular disease, characterized in that selected from the group consisting of hypertension, pulmonary arterial hypertension, atherosclerosis, angina pectoris, myocardial infarction, ischemic cerebrovascular disease, arteriosclerosis, and mesenteric sclerosis;
(ii) scleroderma, rheumatoid arthritis, Crohn's disease, ulcerative colitis, myelofibrosis, pulmonary fibrosis, hepatic fibrosis, Liver cirrhosis, kidney fibrosis, glomerulosclerosis, myofibrosis, cardiac fibrosis, interstitial fibrosis, pancreatic fibrosis, spleen fibrosis, mesenteric fibrosis, vascular fibrosis, skin fibrosis , Eye fibrosis, macular degeneration, joint fibrosis, thyroid fibrosis, endocardial fibrosis, peritoneal fibrosis, peritoneal fibrosis, progressive mass fibrosis, myogenic systemic fibrosis, systemic lupus erythematosus), hereditary fibrosis, infectious fibrosis, irritant fibrosis, fibrosis due to chronic autoimmunity, fibrosis due to antigen incompatibility during organ transplantation, fibrotic complications during surgery, hyperlipidemia By fibrosis, fibrosis caused by obesity, diabetes, fibrosis, fibrotic diseases being selected from the group consisting of occlusion due to fibrosis and fibrosis during stenting due to high blood pressure;
(iii) inflammatory diseases; or
(iv) a pharmaceutical composition characterized by Alport syndrome.
delete According to claim 1, wherein the compound of Formula 1-1
4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid;
4-{[[2- (4-ethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;
4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-propoxyphenyl) ethyl] amino} methyl) benzoic acid;
4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-isopropoxyphenyl) ethyl] amino} methyl) benzoic acid;
4-({(7-fluorobenzo [d] thiazol-2-yl) [2- (4-isobutoxyphenyl) ethyl] amino} methyl) benzoic acid;
4-{[[2- (4-cyclopropylmethoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;
4-{[(7-fluorobenzo [d] thiazol-2-yl) (2-phenylethyl) amino] methyl} benzoic acid;
4-{[[2- (4-cyclobutylmethoxyphenyl) ethyl]-(7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid;
4-{[[2- (4-sec-butoxyphenyl) ethyl] (7-fluorobenzo [d] thiazol-2-yl) amino] methyl} benzoic acid; And
4-({(7-chloro-benzo [d] thiazol-2-yl) [2- (4-methoxyphenyl) ethyl] amino} methyl) benzoic acid
Pharmaceutical composition, characterized in that selected from the group consisting of.

delete delete delete delete delete The method of claim 1, wherein the inflammatory disease is autoimmune disease, inflammatory bowel disease, dermatitis, diabetic eye disease, peritonitis, osteomyelitis, roditis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, rhinitis, sinusitis, otitis media, Pneumonia, gastritis, enteritis, cystic fibrosis, stroke, bronchitis, bronchiolitis, hepatitis, nephritis, arthritis, neuritis, gout, spondylitis, Reiter syndrome, nodular polyarteritis, vasculitis, Lou Gehrig's disease, Wegener's granulomatosis, hypercytokinemia ), Rheumatic polymyalgia, arterial cell arteritis, calcium crystalline arthritis, pseudogout, non-articular rheumatoid arthritis, bursitis, hay fever, epicondylitis, neuropathic joint disease (Charcot's joint), hemarthrosis, henoch Supine purpura, hypertrophic osteoarthritis, multicardiac reticulocytoma, surcoilosis, hemochromatosis, sickle cell disease and other hemochromatosis, hyperlipidemia Hyperemia, hypomagglobulinemia, hyperparathyroidism, acromegaly, familial Mediterranean fever, Behatt's disease, systemic lupus erythematosus, recurrent fever, psoriasis, multiple sclerosis, sepsis, septic shock, acute respiratory distress syndrome, multiple organ failure, A pharmaceutical composition, characterized in that it is selected from the group consisting of chronic obstructive pulmonary disease, acute lung injury and broncho-pulmonary dysplasia.
The method of claim 9, wherein the autoimmune disease is rheumatoid arthritis, systemic scleroderma, systemic lupus erythematosus, psoriasis, asthma, ulcerative colitis, Beche's disease, Crohn's disease, multiple sclerosis, dermatitis, collagen disease, vasculitis, arthritis, granulomatosis Pharmaceutical composition, characterized in that selected from the group consisting of organ-specific autoimmune lesions, ulcerative colitis and graft versus host disease.

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