Therapeutic or prophylactic agent for abnormal angiogenesis and its applications

Abstract

To provide a novel therapeutic or prophylactic agent for abnormal angiogenesis and its applications.SOLUTION: A therapeutic or prophylactic agent for abnormal angiogenesis is used that contains a histone acetyltransferase inhibitor as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a therapeutic or preventive agent for abnormal angiogenesis and use thereof. [Background technology]

[0002] Angiogenesis can occur in vivo under both physiological and pathological conditions. Under physiological conditions, for example, normal angiogenesis occurs in fetal tissues (e.g., retina), which results in the formation of normal tissues. On the other hand, under pathological conditions, for example, abnormal angiogenesis occurs in disease-affected areas (e.g., retinal ischemic disease, cancer, retinopathy of prematurity, age-related macular degeneration, diabetic macular edema, pathological myopia, macular edema associated with retinal vein occlusion, and neovascular glaucoma), which results in the onset or aggravation of the disease.

[0003] Examples of abnormal angiogenesis include the formation of ischemic areas (in other words, the formation of areas where blood vessels are not formed) and the formation of abnormal blood vessels (in other words, the formation of blood vessels that run in an uncontrolled manner).

[0004] The formation of ischemic area and abnormal blood vessels may occur alone or in combination. For example, in age-related macular degeneration and pathological myopia, abnormal blood vessels are formed alone. On the other hand, in retinal ischemic disease, cancer, retinopathy of prematurity, diabetic macular edema, macular edema associated with retinal vein occlusion, and neovascular glaucoma, ischemic area is formed first, and then abnormal blood vessels are formed.

[0005] Anti-VEGF drugs are currently used as therapeutic agents for abnormal angiogenesis (see, for example, Non-Patent Documents 1 and 2). Non-Patent Documents 1 and 2 describe the use of aflibercept, bevacizumab, ranibizumab, and faricimab for the treatment of diabetic macular edema. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] N Eng J Med., 2015 March 26; 372(13): 1193-1203, "Aflibercept, Bevacizumab, or Ranibizumab for Diabetic Macular Edema" [Non-Patent Document 2] Charles C Wykoff et al., Lancet., 2022 Feb 19; 399(10326): 741-755, "Efficacy, durability, and safety of intravitreal faricimab with extended dosing up to every 16 weeks in patients with diabetic macular oedema(YOSEMITE and RHINE): two randomized, double-masked, phase 3 trials" Summary of the Invention [Problem to be solved by the invention]

[0007] However, the above-mentioned conventional techniques have a problem in that the therapeutic effect on abnormal angiogenesis is insufficient.

[0008] For example, the above-mentioned conventional technologies have room for improvement in that: (i) they only have the effect of temporarily reducing abnormal angiogenesis, which raises concerns about the need for frequent administration; (ii) they affect both normal and abnormal angiogenesis, which raises concerns about side effects; and (iii) anti-VEGF drugs inhibit angiogenesis, which raises concerns about the inability to achieve a sufficient therapeutic effect against the formation of ischemic areas.

[0009] An object of one aspect of the present invention is to provide a novel agent for treating or preventing abnormal angiogenesis, and use thereof. [Means for solving the problem]

[0010] One aspect of the present invention is the following: [1] A therapeutic or preventive agent for abnormal angiogenesis, comprising a histone acetyltransferase inhibitor as an active ingredient.

[0011] [2] The therapeutic or preventive agent for abnormal angiogenesis according to [1], wherein the histone acetyltransferase inhibitor targets one or more substances selected from the group consisting of HAT1, NAA60, GCN5, PCAF, TIP60, MOZ, MORF, HBO1, MOF, TAF1, TIFIIIC90, SRC1, SCR3, P600, CLOCK, NAT10, P300, CBP, and BRD4.

[0012] [3] The agent for treating or preventing abnormal angiogenesis according to [1], wherein the histone acetyltransferase inhibitor is a compound represented by the following chemical formula 1, a derivative thereof, or a salt thereof:

[0013] [ka]

[0014] [4] The therapeutic or preventive agent for abnormal angiogenesis according to [1], wherein the abnormal angiogenesis is the formation of an ischemic area and / or the formation of abnormal blood vessels.

[0015] [5] The therapeutic or preventive agent for abnormal angiogenesis according to any one of [1] to [4], wherein the abnormal angiogenesis is abnormal angiogenesis associated with retinal ischemic disease, cancer, retinopathy of prematurity, age-related macular degeneration, diabetic macular edema, pathological myopia, macular edema associated with retinal vein occlusion, or neovascular glaucoma.

[0016] [6] Use of a histone acetyltransferase inhibitor for the manufacture of an agent for treating or preventing abnormal angiogenesis.

[0017] [7] An administration step of administering a candidate substance for a therapeutic or preventive agent for abnormal angiogenesis to a non-human model organism that has developed abnormal angiogenesis; a measuring step of measuring, in the non-human model organism after the administration step, (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene; A screening method for a therapeutic or preventive agent for abnormal angiogenesis, comprising a determination step of determining whether the candidate substance is a therapeutic or preventive agent for abnormal angiogenesis by comparing the expression level of each gene measured in the measurement step with a control expression level.

[0018] [8] A method for obtaining data for diagnosing abnormal angiogenesis, comprising a measuring step of measuring (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene in a biological sample collected from a subject.

[0019] [9] A marker for diagnosing abnormal angiogenesis, comprising: (i) the Ang2 gene; and (ii) at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene. Effect of the Invention

[0020] According to one aspect of the present invention, it is possible to provide a new agent for treating or preventing abnormal angiogenesis, and use thereof. [Brief description of the drawings]

[0021] [Figure 1] 101 is an image showing the effect of a therapeutic agent and a preventive agent for abnormal angiogenesis according to an embodiment of the present invention, and 102 is a graph showing the effect of a therapeutic agent and a preventive agent for abnormal angiogenesis according to an embodiment of the present invention. [Diagram 2] 201 is an image showing the effect of a therapeutic agent and a preventive agent for abnormal angiogenesis according to an embodiment of the present invention, and 202 is a graph showing the effect of a therapeutic agent and a preventive agent for abnormal angiogenesis according to an embodiment of the present invention. [Diagram 3] Graphs 301 to 312 show changes in the expression levels of various genes during the development and healing process of abnormal angiogenesis in an example of the present invention. [Figure 4] Graphs 401 to 410 show changes in the expression levels of various genes during the development and healing process of abnormal angiogenesis in an example of the present invention. [Diagram 5] 1 is a graph showing the change in the expression level of VEGF-A during the development and healing process of abnormal angiogenesis in an example of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] An embodiment of the present invention will be described below, but the present invention is not limited thereto. The present invention is not limited to each configuration described below, and various modifications are possible within the scope of the claims, and embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention. In addition, all academic literature and patent documents described in this specification are incorporated herein by reference. In addition, unless otherwise specified in this specification, "A to B" representing a numerical range means "A or more, B or less."

[0023] The present invention can also contribute to achieving Goal 3 of the Sustainable Development Goals (SDGs) advocated by the United Nations, which is to "Ensure good health and promote well-being for all."

[0024] 1. Technical Concept of the Present Invention In vivo, normal angiogenesis occurs under physiological conditions, whereas abnormal angiogenesis occurs under pathological conditions. To form a normal organism, it is necessary to suppress abnormal angiogenesis while allowing normal angiogenesis to occur.

[0025] The present inventors have conducted research based on the original hypothesis that a substance capable of returning the pathological conditions that cause abnormal angiogenesis to the physiological conditions that cause normal angiogenesis would be useful as a new therapeutic or preventive agent for abnormal angiogenesis.

[0026] Specifically, the present inventors screened for new candidate substances for treating and preventing abnormal angiogenesis based on the unique hypothesis that substances that target proteins that control the expression of genes subject to epigenetic expression control are substances that can return pathological conditions that cause abnormal angiogenesis to physiological conditions that cause normal angiogenesis.

[0027] Numerous proteins are known to date that regulate the expression of genes that are subject to epigenetic expression control. The present inventors have found that, among these proteins, substances that target histone acetyltransferases in particular are useful as new therapeutic and preventive agents for abnormal angiogenesis, and have thus completed the present invention.

[0028] 2. Therapeutic or preventive agents for abnormal angiogenesis An agent for treating or preventing abnormal angiogenesis according to one embodiment of the present invention contains a histone acetyl transferase (HAT) inhibitor as an active ingredient.

[0029] The therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention has the following advantages: (i) since it has the effect of fundamentally treating or preventing abnormal angiogenesis, there is no need to worry about frequent administration; (ii) since it does not affect normal angiogenesis, there is no need to worry about side effects; and (iii) since it does not suppress angiogenesis like anti-VEGF drugs, it has a sufficient therapeutic effect on the formation of ischemic areas.

[0030] Histone acetyltransferase is an enzyme that replaces the amino group of a specific lysine residue contained in histone with an acetyl group. This replacement partially loosens the chromatin structure formed by DNA and histone, making it easier for genes near the site where the histone is acetylated to be expressed. Histone acetyltransferase inhibitors have the effect of suppressing the expression of specific genes by inhibiting the above-mentioned process. Therefore, it is believed that the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention acts on the expression of genes that are subject to epigenetic expression control.

[0031] As used herein, the term "therapeutic agent" refers to a drug that provides a therapeutic effect. The above therapeutic effect refers to, but is not limited to, the effects exemplified below.

[0032] (1) The severity of abnormal angiogenesis is reduced compared to when the therapeutic agent is not administered.

[0033] (2) preventing an increase in severity or progression of abnormal angiogenesis compared to when the therapeutic agent is not administered;

[0034] (3) reducing the rate at which the severity of abnormal angiogenesis increases or progresses, compared to when the therapeutic agent is not administered;

[0035] As used herein, the term "prophylactic agent" refers to a drug that provides a prophylactic effect. The prophylactic effect includes, but is not limited to, the effects exemplified below.

[0036] (1) Preventing the occurrence of abnormal angiogenesis or reducing the risk compared to when the prophylactic agent is not administered.

[0037] (2) Preventing or reducing the risk of recurrence of abnormal angiogenesis compared to when the prophylactic agent is not administered.

[0038] (3) preventing or reducing the risk of symptoms of abnormal angiogenesis compared to when the prophylactic agent is not administered;

[0039] The abnormal angiogenesis may be systemic or local.

[0040] The above histone acetyltransferase inhibitors include HAT1 (official name: Histone Acetyltransferase 1), NAA60 (official name: N-Alpha-Acetyltransferase 60), GCN5 (official name: General Control Of Amino Acid Synthesis 5), PCAF (official name: P300 / CBP-Associated Factor), TIP60 (official name: Tat-Interactive Protein 60kDa), MOZ (official name: Monocytic Leukemia Zinc Finger), MORF (official name: Monocytic Leukemia Zinc Finger Protein-Related Factor), HBO1 (official name: Histone Acetyltransferase Binding To ORC1), MOF (official name: Males Absent On The First), TAF1 (official name: TATA-Box Binding Protein Associated Factor) 1), TIFIIIC90 (official name: Histone Acetyltransferase It is preferable that the targeting agent targets one or more substances, preferably two or more substances, and most preferably three or more substances selected from the group consisting of TIFIIIC90), SRC1 (official name: Steroid Receptor Coactivator 1), SCR3 (official name: Steroid Receptor Coactivator 3), P600 (official name: Histone Acetyltransferase P600), CLOCK (official name: Histone Acetyltransferase CLOCK), NAT10 (official name: N-Acetyltransferase 10), P300 (official name: Histone Acetyltransferase P300), CBP (official name: CREB-Binding Protein), and BRD4 (official name: Bromodomain Protein 4).The histone acetyltransferase inhibitor preferably targets one or more substances, preferably two or more substances, most preferably three substances selected from the group consisting of P300, CBP, and BRD4. With this configuration, abnormal angiogenesis can be effectively treated or prevented.

[0041] The histone acetyltransferase inhibitor is preferably a compound represented by the following chemical formula 1, a derivative thereof, or a salt thereof. With this composition, abnormal angiogenesis can be effectively treated or prevented.

[0042] [ka]

[0043] The compound represented by the above chemical formula 1 is a compound commercially available as "ISOX DUAL". This compound is known to target P300, CBP, and BRD4.

[0044] The above-mentioned derivatives and salts may have any structure as long as they have inhibitory activity against histone acetyltransferase. Whether or not a specific substance has inhibitory activity against histone acetyltransferase can be determined according to a known method. For example, after mixing a specific substance with histone acetyltransferase, it can be examined whether or not the amino group of a specific lysine residue contained in histone is replaced with an acetyl group by the histone acetyltransferase. In this test, if the amino group of a specific lysine residue contained in histone is not replaced with an acetyl group by the histone acetyltransferase, it can be determined that the specific substance has inhibitory activity against histone acetyltransferase.

[0045] As used herein, the term "derivative" refers to a group of compounds that are produced by replacing a portion of a molecule of a specific compound with another functional group or another atom.

[0046] Examples of the other functional groups include an alkyl group, an alkoxy group, an alkylthio group, an aryl group, an aryloxy group, an arylthio group, an arylalkyl group, an arylalkoxy group, an arylalkylthio group, an arylalkenyl group, an arylalkynyl group, an allyl group, an amino group, a substituted amino group, a silyl group, a substituted silyl group, a silyloxy group, a substituted silyloxy group, an arylsulfonyloxy group, an alkylsulfonyloxy group, a nitro group, etc. Examples of the other atoms include a carbon atom, a hydrogen atom, an oxygen atom, a nitrogen atom, a sulfur atom, a phosphorus atom, a halogen atom, etc.

[0047] In this specification, the term "salt" refers to any physiologically acceptable salt that can be administered to a subject as a pharmaceutical, and is not limited thereto. Examples of the salt include alkali metal salts (potassium salt, etc.), alkaline earth metal salts (calcium salt, magnesium salt, etc.), ammonium salts, organic base salts (trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.), organic acid salts (acetate salt, maleate salt, tartrate salt, methanesulfonate salt, benzenesulfonate salt, formate salt, toluenesulfonate salt, trifluoroacetate salt, etc.), inorganic acid salts (hydrochloride salt, hydrobromide salt, sulfate salt, phosphate salt, etc.), etc.

[0048] The type of abnormal angiogenesis is not limited. The abnormal angiogenesis may be the formation of an ischemic region (in other words, the formation of a region where blood vessels are not formed) and / or the formation of abnormal blood vessels (in other words, the formation of blood vessels running in a disorderly manner). The abnormal angiogenesis may be abnormal angiogenesis including the formation of an ischemic region, abnormal angiogenesis including the formation of abnormal blood vessels, or abnormal angiogenesis including both the formation of an ischemic region and the formation of abnormal blood vessels.

[0049] The therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention uses a mechanism of action completely different from that of conventional anti-VEGF drugs and does not suppress angiogenesis. Therefore, as shown in the examples described below, the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention can treat and prevent not only the formation of abnormal blood vessels but also the formation of ischemic regions.

[0050] The disease in which the abnormal angiogenesis occurs is not limited. The abnormal angiogenesis may be associated with retinal ischemic disease, cancer, retinopathy of prematurity, age-related macular degeneration, diabetic macular edema, pathological myopia, macular edema associated with retinal vein occlusion, or neovascular glaucoma. The abnormal angiogenesis may be associated with retinal ischemic disease, cancer, retinopathy of prematurity, diabetic macular edema, macular edema associated with retinal vein occlusion, or neovascular glaucoma, which are accompanied by the formation of ischemic areas. The abnormal angiogenesis may be associated with age-related macular degeneration or pathological myopia, which are accompanied by the formation of abnormal blood vessels and are not accompanied by the formation of ischemic areas.

[0051] The therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention can treat or prevent abnormal angiogenesis associated with the above-mentioned diseases. Furthermore, the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention can treat or prevent increased vascular permeability and ischemia caused by the above-mentioned diseases.

[0052] The abnormal angiogenesis treatment or prevention agent according to one embodiment of the present invention can be administered to a subject by any administration route. Examples of the administration route include parenteral administration (e.g., ophthalmic administration, intraconjunctival administration, intravitreal administration, subconjunctival administration, subtenon administration, intraperitoneal administration), oral administration, transdermal administration, transmucosal administration, and intravenous administration. Therefore, the dosage form of the abnormal angiogenesis treatment or prevention agent according to one embodiment of the present invention can be eye drops, oral medication, topical medication, injection, inhalant, eye drops, etc.

[0053] The amount of an active ingredient contained in the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention is not particularly limited, and may be, for example, 0.00001% by mass to 100% by mass, 0.0001% by mass to 100% by mass, 0.001% by mass to 100% by mass, 0.01% by mass to 100% by mass, 0.1% by mass to 100% by mass, or 0.1% by mass to 100% by mass. The content may be 0.1% to 95% by mass, 0.1% to 90% by mass, 0.1% to 80% by mass, 0.1% to 70% by mass, 0.1% to 60% by mass, 0.1% to 50% by mass, 0.1% to 40% by mass, 0.1% to 30% by mass, 0.1% to 20% by mass, or 0.1% to 10% by mass.

[0054] When the agent for treating or preventing abnormal angiogenesis according to one embodiment of the present invention is in a liquid form, the amount of the active ingredient contained in the agent for treating or preventing abnormal angiogenesis according to one embodiment of the present invention is not particularly limited and may be, for example, 0.1 μM to 1 mM, 1 μM to 500 μM, 1 μM to 300 μM, or 10 μM to 150 μM.

[0055] The agent for treating or preventing abnormal angiogenesis according to one embodiment of the present invention may contain ingredients other than the above-mentioned active ingredients.

[0056] The components other than the above-mentioned active ingredient are not particularly limited, and may be, for example, a buffering agent, a pH adjusting agent, an isotonicity agent, a preservative, an antioxidant, a high molecular weight polymer, an excipient, a solvent, an antibacterial agent, and the like.

[0057] Examples of the buffer include phosphoric acid or a phosphate salt, boric acid or a borate salt, citric acid or a citrate salt, acetic acid or an acetate salt, carbonic acid or a carbonate salt, tartaric acid or a tartrate salt, ε-aminocaproic acid, and trometamol. Examples of the phosphate salt include sodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate. Examples of the borate salt include borax, sodium borate, and potassium borate. Examples of the citrate salt include sodium citrate, disodium citrate, and trisodium citrate. Examples of the acetate salt include sodium acetate and potassium acetate. Examples of the carbonate salt include sodium carbonate and sodium bicarbonate. Examples of the tartrate salt include sodium tartrate and potassium tartrate.

[0058] Examples of the pH adjuster include hydrochloric acid, phosphoric acid, citric acid, acetic acid, sodium hydroxide, and potassium hydroxide.

[0059] Examples of the isotonicity agent include ionic isotonicity agents (eg, sodium chloride, potassium chloride, calcium chloride, magnesium chloride) and non-ionic isotonicity agents (eg, glycerin, propylene glycol, sorbitol, mannitol).

[0060] Examples of the preservative include benzalkonium chloride, benzalkonium bromide, benzethonium chloride, sorbic acid, potassium sorbate, methyl parahydroxybenzoate, propyl parahydroxybenzoate, and chlorobutanol.

[0061] Examples of the antioxidant include ascorbic acid, tocopherol, dibutylhydroxytoluene, butylhydroxyanisole, sodium erythorbate, propyl gallate, and sodium sulfite.

[0062] Examples of the high molecular weight polymer include methyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropyl methyl cellulose phthalate, carboxymethyl ethyl cellulose, cellulose acetate phthalate, polyvinylpyrrolidone, polyvinyl alcohol, carboxyvinyl polymer, polyethylene glycol, and atelocollagen.

[0063] Examples of the excipient include lactose, sucrose, D-mannitol, xylitol, sorbitol, erythritol, starch, and crystalline cellulose.

[0064] Examples of the solvent include water, physiological saline, and alcohol.

[0065] Examples of the antibacterial agents include β-lactam, aminoglycoside, tetracycline, lincomycin, chloramphenicol, macrolide, ketolide, polypeptide, and glycopeptide antibiotics; and pyridonecarboxylic acid (quinolone), new quinolone, oxazolidinone, and sulfa drug-based synthetic antibacterial agents.

[0066] The amount of ingredients other than the active ingredient contained in the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention is not particularly limited, and may be, for example, 0% by mass to 99.99999% by mass, 0% by mass to 99.9999% by mass, 0% by mass to 99.9999% by mass, 0% by mass to 99.9999% by mass, 0% by mass to 99.999% by mass, 0% by mass to 99.99% by mass, 5% by mass to 99. It may be 9% by mass, 10% by mass to 99.9% by mass, 20% by mass to 99.9% by mass, 30% by mass to 99.9% by mass, 40% by mass to 99.9% by mass, 50% by mass to 99.9% by mass, 60% by mass to 99.9% by mass, 70% by mass to 99.9% by mass, 80% by mass to 99.9% by mass, or 90% by mass to 99.9% by mass.

[0067] The administration interval of the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention is not limited, and may be, for example, once every 12 hours, once a day, once every two days, once every three days, once every four days, or once every five days. The administration period of the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention is not limited, and may be, for example, 3 days, 5 days, 7 days, 10 days, 15 days, or 30 days. The therapeutic or preventive agent according to one embodiment of the present invention can fundamentally treat or prevent abnormal angiogenesis, and therefore does not require excessively frequent administration.

[0068] The subjects of administration of the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention are not limited, and examples thereof include humans and non-human animals (e.g., livestock, pets, and laboratory animals). Examples of non-human animals include monkeys, chimpanzees, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, mice, and rats.

[0069] 3. Use of histone acetyltransferase inhibitors The matters explained in the above section "2. Agent for treating or preventing abnormal angiogenesis" will not be explained further below.

[0070] A use according to one embodiment of the present invention is the use of a histone acetylase inhibitor for the manufacture of an agent for the treatment or prevention of abnormal angiogenesis.

[0071] The histone acetyltransferase inhibitor is preferably one that targets one or more substances, preferably two or more substances, most preferably three or more substances selected from the group consisting of HAT1, NAA60, GCN5, PCAF, TIP60, MOZ, MORF, HBO1, MOF, TAF1, TIFIIIC90, SRC1, SCR3, P600, CLOCK, NAT10, P300, CBP, and BRD4. The histone acetyltransferase inhibitor is preferably one that targets one or more substances, preferably two or more substances, most preferably three substances selected from the group consisting of P300, CBP, and BRD4.

[0072] The histone acetyltransferase inhibitor is preferably a compound represented by the following chemical formula 1, a derivative thereof, or a salt thereof.

[0073] [ka]

[0074] The type of abnormal angiogenesis is not limited. The abnormal angiogenesis may be the formation of an ischemic region (in other words, the formation of a region where blood vessels are not formed) and / or the formation of abnormal blood vessels (in other words, the formation of blood vessels running in a disorderly manner). The abnormal angiogenesis may be abnormal angiogenesis including the formation of an ischemic region, abnormal angiogenesis including the formation of abnormal blood vessels, or abnormal angiogenesis including both the formation of an ischemic region and the formation of abnormal blood vessels.

[0075] The disease in which the abnormal angiogenesis occurs is not limited. The abnormal angiogenesis may be associated with retinal ischemic disease, cancer, retinopathy of prematurity, age-related macular degeneration, diabetic macular edema, pathological myopia, macular edema associated with retinal vein occlusion, or neovascular glaucoma. The abnormal angiogenesis may be associated with retinal ischemic disease, cancer, retinopathy of prematurity, diabetic macular edema, macular edema associated with retinal vein occlusion, or neovascular glaucoma, which are accompanied by the formation of ischemic areas. The abnormal angiogenesis may be associated with age-related macular degeneration or pathological myopia, which are accompanied by the formation of abnormal blood vessels and are not accompanied by the formation of ischemic areas.

[0076] The method for producing the agent for treating or preventing abnormal angiogenesis is not limited, and any method generally used in the art may be used. For example, the agent for treating or preventing abnormal angiogenesis may be produced by mixing the above-mentioned active ingredient with, if necessary, an ingredient other than the above-mentioned active ingredient.

[0077] 4. Screening method for therapeutic or preventive agents for abnormal angiogenesis A method for screening a therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention includes the steps of: administering a candidate substance for a therapeutic or preventive agent for abnormal angiogenesis to a non-human model organism having developed abnormal angiogenesis; and measuring, in the non-human model organism after the administration step, (i) the expression level of the Ang2 (formal name: Angiopoietin 2) gene, and (ii) the expression levels of the Adm (formal name: Adrenomedullin) gene, the Anxa2 (formal name: Annexin A2) gene, the Apln (formal name: Apelin) gene, the Cd34 (formal name: Cluster of differentiation 34) gene, the Cxcr4 (formal name: CXC Motif Chemokine Receptor 4) gene, the Edn1 (formal name: Endothelin 1) gene, the Ednra (formal name: Endothelin Receptor Type A) gene, the Eng (formal name: Endoglin) gene, the Fn1 (formal name: Fibronectin 1) gene, Hmox1 (official name: Heme oxygenase 1) gene, Itga1 (official name: Integrin Subunit Alpha 1) gene, Loxl2 (official name: Lysyl Oxidase Like 2) gene, Mcam (official name: Melanoma Cell Adhesion Molecule) gene, Nr4a1 (official name: Nuclear Receptor Subfamily 4 Group A Member 1) gene, Serpine1 (official name: Serpin Family E Member 1) gene, Tnfrsf12a (official name: TNF Receptor Superfamily Member 12A) gene, Acta2 (official name: Actin Alpha 2) gene, Anxa3 (official name: Annexin A3) gene, Thbs (official name: Thrombospondin 1) gene, Cdh5 (official name: Cadherin 5) gene, and Esm1 (official name: Endothelial Cell-specific molecules1) The method comprises a measuring step of measuring the expression level of at least one gene (e.g., at least 3, at least 5, at least 8, at least 10, at least 13, at least 15, at least 18, at least 20, or 21) selected from a group consisting of genes, and a determining step of determining whether or not the candidate substance is a therapeutic or preventive agent for abnormal angiogenesis by comparing the expression level of each gene measured in the measuring step with a control expression level.

[0078] The non-human model organism with abnormal angiogenesis is not limited, and for example, an OIR (Oxygen-Induced Retinopathy) model organism (e.g., an OIR model mouse) can be used. The OIR model organism can be prepared by exposing a newborn organism to high-oxygen conditions (e.g., 75% by volume oxygen conditions) for a desired period of time, and then exposing the organism to atmospheric conditions for a desired period of time. Details of the method for preparing the OIR model organism are disclosed in, for example, the literature "Lois EH Smith et al., "Oxygen-Induced Retinopathy in the Mouse" Investigative Ophthalmology and Visual Science, January 1994, Vol.35, No.1, p101-111". In addition, tissues or cells collected from the non-human model organism can also be used as the non-human model organism with abnormal angiogenesis.

[0079] The candidate substance for the therapeutic or preventive agent for neovascularization is not limited, and may be, for example, an organic compound or an inorganic compound.

[0080] In the administration step, the candidate substance is administered to the non-human model organism. The specific configuration of the administration step is not limited, and for example, (i) the candidate substance may be administered to the non-human model organism itself (e.g., parenteral administration (e.g., ophthalmic administration, intraconjunctival administration, intravitreal administration, subconjunctival administration, sub-Tenon administration, intraperitoneal administration), oral administration, transdermal administration, transmucosal administration, intravenous administration), or (ii) the candidate substance may be added to a medium in which tissues or cells collected from the non-human model organism are cultured.

[0081] In the measuring step, (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene selected from the above-mentioned 21 genes are measured.

[0082] It is known that angiopoietin 2 and VEGF (vascular endothelial growth factor) are necessary for blood vessel elongation. As shown in the examples described later, when abnormal angiogenesis occurs, the expression level of VEGF increases. However, the increase in the expression level of VEGF does not decrease with the cure of abnormal angiogenesis. On the other hand, when abnormal angiogenesis occurs, the expression levels of the above-mentioned 21 genes increase. The increase in the expression levels of the above-mentioned 21 genes decreases with the cure of abnormal angiogenesis. This suggests that both (i) a signal transduction system involving VEGF (VEGF-dependent signal transduction system) and (ii) a signal transduction system involving at least one gene selected from the above-mentioned 21 genes (VEGF-independent signal transduction system) are involved in the cure of abnormal angiogenesis. In other words, this suggests that the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention exerts its effect through both the VEGF-dependent signal transduction system and the VEGF-independent signal transduction system. Therefore, according to the above-mentioned measurement process, screening is performed based on the two indicators (i) and (ii), so that a therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention can be efficiently screened.

[0083] In the measurement step, the expression level of each gene may be measured based on the expression level of mRNA, or may be measured based on the expression level of protein. When the expression level of each gene is measured based on the expression level of mRNA, the measurement step can be performed based on a known method such as RT-PCR or Northern blotting. When the expression level of each gene is measured based on the expression level of protein, the measurement step can be performed based on a known method such as ELISA or Western blotting.

[0084] In the determination step, the expression level of each gene measured in the measurement step is compared with a control expression level to determine whether or not the candidate substance is a therapeutic or preventive agent for abnormal angiogenesis.

[0085] The control expression level can be, for example, the expression level of each gene previously measured in a non-human model organism that has developed abnormal angiogenesis, and / or the expression level of each gene previously measured in a non-human model organism that has not developed abnormal angiogenesis.

[0086] The expression level of each gene measured in the measurement step is designated as "A", and the expression level of each gene previously measured in the non-human model organism with abnormal angiogenesis is designated as "B". When the value of "A / B" is, for example, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less, it can be determined that the candidate substance is a therapeutic or preventive agent for abnormal angiogenesis. The lower limit of the value of "A / B" is not limited, and may be, for example, 0.001 or 0.01.

[0087] The expression level of each gene measured in the measuring step is designated as "A," and the expression level of each gene previously measured in the non-human model organism not suffering from abnormal angiogenesis is designated as "C." When the "A / C" value is, for example, 0.5 to 1.5, 0.6 to 1.4, 0.7 to 1.3, 0.8 to 1.2, 0.9 to 1.1, or approximately 1.0, the candidate substance can be determined to be a therapeutic or preventive agent for abnormal angiogenesis.

[0088] 5. Method for obtaining data for diagnosing abnormal angiogenesis, and marker for diagnosing abnormal angiogenesis A method for obtaining data for diagnosing abnormal angiogenesis according to one embodiment of the present invention includes a measurement step of measuring, in a biological sample collected from a subject, (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene (e.g., at least 3, at least 5, at least 8, at least 10, at least 13, at least 15, at least 18, at least 20, or 21) selected from the group consisting of Adm gene, Anxa2 gene, Apln gene, Cd34 gene, Cxcr4 gene, Edn1 gene, Ednra gene, Eng gene, Fn1 gene, Hmox1 gene, Itga1 gene, Loxl2 gene, Mcam gene, Nr4a1 gene, Serpine1 gene, Tnfrsf12a gene, Acta2 gene, Anxa3 gene, Thbs gene, Cdh5 gene, and Esm1 gene.

[0089] The specific configuration of the above measuring step may be the same as the measuring step in the above-mentioned [4. Screening method for therapeutic or preventive agents for abnormal angiogenesis].

[0090] The biological sample collected from the subject is not limited, and may be, for example, cells or tissue collected from the affected area of ​​a disease.

[0091] By comparing the expression level of each gene measured in the measuring step with a control expression level, it is possible to determine whether or not the subject suffers from abnormal angiogenesis.

[0092] The control expression level can be, for example, the expression level of each gene previously measured in patients who have developed abnormal angiogenesis, and / or the expression level of each gene previously measured in healthy individuals who have not developed abnormal angiogenesis.

[0093] The expression level of each gene measured in the measurement step is designated as "A", and the expression level of each gene previously measured in a healthy subject not suffering from abnormal angiogenesis is designated as "B". When the value of "A / B" is, for example, 1.5 or more, 2.0 or more, 3.0 or more, 5.0 or more, or 10.0 or more, it can be determined that the subject suffers from abnormal angiogenesis. The upper limit of the value of "A / B" is not limited, and may be, for example, 10.0 or 20.0.

[0094] The expression level of each gene measured in the measurement step is designated as "A", and the expression level of each gene previously measured in the patient who has developed abnormal angiogenesis is designated as "C". When the value of "A / C" is, for example, 1.0 or more, it can be determined that the subject suffers from abnormal angiogenesis. The upper limit of the value of "A / C" is not limited, and may be, for example, 5.0 or 10.0.

[0095] As described above, in the method for acquiring data for diagnosing abnormal angiogenesis according to one embodiment of the present invention, abnormal angiogenesis can be diagnosed based on the expression levels of 22 types of genes. In other words, the above 22 types of genes can be used as markers for diagnosing abnormal angiogenesis.

[0096] Therefore, a marker for diagnosing abnormal angiogenesis in one embodiment of the present invention includes (i) the Ang2 gene, and (ii) at least one gene (e.g., at least 3, at least 5, at least 8, at least 10, at least 13, at least 15, at least 18, at least 20, or 21) selected from the group consisting of Adm gene, Anxa2 gene, Apln gene, Cd34 gene, Cxcr4 gene, Edn1 gene, Ednra gene, Eng gene, Fn1 gene, Hmox1 gene, Itga1 gene, Loxl2 gene, Mcam gene, Nr4a1 gene, Serpine1 gene, Tnfrsf12a gene, Acta2 gene, Anxa3 gene, Thbs gene, Cdh5 gene, and Esm1 gene. EXAMPLES

[0097] The present invention will be described in more detail below based on examples, but the present invention is not limited to the following examples.

[0098] <1. Drug screening (intraperitoneal administration)> Various drug candidates were intraperitoneally administered to OIR model mice that had developed abnormal angiogenesis, and their effects on abnormal angiogenesis were confirmed.

[0099] Seven-day-old (P7) mice were reared under 75% oxygen by volume for 5 days until they reached 12-day-old (P12) age. The 12-day-old (P12) mice were then reared under atmospheric conditions. By switching rearing conditions from high oxygen concentration to low oxygen concentration, abnormal angiogenesis (formation of ischemic areas (in other words, formation of areas where blood vessels are not formed) and formation of abnormal blood vessels (in other words, formation of blood vessels running in a disorderly manner)) was induced in the retina.

[0100] Various drug candidates were intraperitoneally administered to the above-mentioned mice once a day for 5 days from postnatal day 12 (P12) to postnatal day 17 (P17). ISOX DUAL (concentration: 150 μM) was used as one of the drug candidates. As a control test, mice that were not intraperitoneally administered with the drug candidate were prepared.

[0101] At 17 days of age, retinas were harvested from the mice and abnormal neovascularization on the retinas was observed under a microscope. Based on the images, the area of ​​the entire retina, the area of ​​the ischemic area, and the area of ​​the abnormal vascular area were calculated.

[0102] The image of the retina is shown in FIG. 1, 101. As is clear from the image, in the control mice, the formation of ischemic areas and abnormal blood vessels was observed in a wide area on the retina. On the other hand, in the mice administered ISOX DUAL, the formation of ischemic areas and abnormal blood vessels was observed only in a narrow area on the retina.

[0103] In FIG. 1, 102, the ratio of the area of ​​abnormal vascular area to the total area of ​​retina and the ratio of the area of ​​ischemic area to the total area of ​​retina are shown in the graph. As can be seen from the graph, in the control test mice, the formation of ischemic area and the formation of abnormal blood vessels were observed in a wide area on the retina. On the other hand, in the mice administered with ISOX DUAL, the formation of ischemic area and the formation of abnormal blood vessels were observed only in a narrow area on the retina.

[0104] <2. Intravitreal administration of ISOX DUAL> ISOX DUAL was administered intravitreally to OIR model mice that had developed abnormal angiogenesis, and its effect on abnormal angiogenesis was confirmed.

[0105] Seven-day-old (P7) mice were reared under 75% oxygen by volume for 5 days until they reached 12-day-old (P12) age. The 12-day-old (P12) mice were then reared under atmospheric conditions. By switching rearing conditions from high oxygen concentration to low oxygen concentration, abnormal angiogenesis (formation of ischemic areas (in other words, formation of areas where blood vessels are not formed) and formation of abnormal blood vessels (in other words, formation of blood vessels running in a disorderly manner)) was induced in the retina.

[0106] The mice were administered ISOX DUAL (concentration: 10 μM) intravitreally once at postnatal day 12 (P12) in the right eye. As a control, the same mice were administered PBS intravitreally once at postnatal day 12 (P12) in the left eye.

[0107] At 17 days of age, retinas were harvested from the mice and abnormal neovascularization on the retinas was observed under a microscope. Based on the images, the area of ​​the entire retina, the area of ​​the ischemic area, and the area of ​​the abnormal vascular area were calculated.

[0108] The image of the retina is shown in 201 of Figure 2. As is clear from the image, in the control mice, the formation of ischemic areas and abnormal blood vessels was observed in a wide area on the retina. On the other hand, in the mice administered ISOX DUAL, the formation of ischemic areas and abnormal blood vessels was observed only in a narrow area on the retina.

[0109] 202 of FIG. 2 shows the ratio of the area of ​​abnormal vascular area to the total area of ​​retina, and the ratio of the area of ​​ischemic area to the total area of ​​retina. As can be seen from this graph, in the control test mice, the formation of ischemic area and the formation of abnormal blood vessels were observed in a wide area on the retina. On the other hand, in the mice administered with ISOX DUAL, the formation of ischemic area and the formation of abnormal blood vessels were observed only in a narrow area on the retina.

[0110] <3. Screening of genes related to the occurrence of abnormal angiogenesis and the action and effect of drugs> A gene whose expression level increases with the occurrence of abnormal angiogenesis and whose expression level decreases when a drug exerts its effect is extremely useful as a screening agent for treating or preventing abnormal angiogenesis, a diagnosis of abnormal angiogenesis, a marker for diagnosing abnormal angiogenesis, etc. Therefore, the present inventors screened for such a gene.

[0111] In this study, we prepared (i) mice that did not develop abnormal angiogenesis and were not administered ISOX DUAL (Control), (ii) mice that developed abnormal angiogenesis and were not administered ISOX DUAL (OIR), and (iii) mice that developed abnormal angiogenesis and were administered ISOX DUAL (ISOX).

[0112] The mice were bred until they were 17 days old, and these mice were used as the above (i) mice (Control) (n=2).

[0113] Seven-day-old (P7) mice were raised under 75% oxygen by volume for 5 days until they reached 12 days of age (P12). The P12 mice were then raised under air conditions until they reached 17 days of age, and these mice were designated as mice (OIR) in (ii) above (n=2).

[0114] A 7-day-old (P7) mouse was raised under 75% oxygen by volume for 5 days until it reached 12 days of age (P12). ISOX DUAL (concentration: 10 μM) was administered intravitreally to the mouse once at 12 days of age (P12). The mouse was then raised under atmospheric conditions until it reached 17 days of age, and was designated as the mouse (ISOX) in (iii) above (n=2).

[0115] Retinas were harvested from each mouse at 17 days of age, and mRNA was purified from the retinas using a commercially available kit. The mRNA was subjected to microarray analysis using a commercially available device (Affimetrix, GCS3000 device) to screen for genes with large variations in expression levels.

[0116] More specifically, genes with large variations in expression levels were screened for, namely, genes whose expression levels varied by 1.5-fold or more between mice (Control) and mice (OIR), and genes whose expression levels varied by 1.5-fold or more between mice (OIR) and mice (ISOX).

[0117] The test results are shown in Figures 3 and 4. As is clear from Figures 3 and 4, significant variations were observed in the expression levels of the Ang2 gene, Adm gene, Anxa2 gene, Apln gene, Cd34 gene, Cxcr4 gene, Edn1 gene, Ednra gene, Eng gene, Fn1 gene, Hmox1 gene, Itga1 gene, Loxl2 gene, Mcam gene, Nr4a1 gene, Serpine1 gene, Tnfrsf12a gene, Acta2 gene, Anxa3 gene, Thbs gene, Cdh5 gene, and Esm1 gene.

[0118] <4. Analysis of signal transduction pathways involved in the treatment and prevention of abnormal angiogenesis> It is known that angiopoietin 2 and VEGF (vascular endothelial growth factor) are necessary for blood vessel growth. Therefore, we confirmed whether the expression pattern of VEGF during the development and healing of abnormal angiogenesis coincided with the expression pattern of the genes screened in <3. Screening of genes related to the development of abnormal angiogenesis and the action and effect of drugs> mentioned above.

[0119] In this study, we prepared (i) mice that did not develop abnormal angiogenesis and were not administered ISOX DUAL (Control), (ii) mice that developed abnormal angiogenesis and were not administered ISOX DUAL (OIR), and (iii) mice that developed abnormal angiogenesis and were administered ISOX DUAL (ISOX).

[0120] The mice were bred until they were 17 days old, and these mice were used as the above (i) mice (Control).

[0121] Seven-day-old (P7) mice were raised under 75% oxygen by volume for 5 days until they reached 12 days of age (P12). The P12 mice were then raised under atmospheric conditions until they reached 17 days of age, and these mice were designated as mice (OIR) in (ii) above.

[0122] A 7-day-old (P7) mouse was raised under 75% oxygen by volume for 5 days until it reached 12 days of age (P12). ISOX DUAL (concentration: 10 μM) was administered intravitreally to the mouse once at 12 days of age (P12). The mouse was then raised under atmospheric conditions until it reached 17 days of age, and was designated as the mouse (ISOX) in (iii) above.

[0123] Retinas were harvested from each mouse at 17 days of age, and mRNA was purified from the retinas using a commercially available kit. VEGF-A mRNA expressed in the retinas of each mouse was quantified by RT-qPCR using the mRNA and VEGF-A-specific primers.

[0124] Figure 5 shows the expression level of VEGF-A in each mouse. As shown in Figure 5, when abnormal angiogenesis occurred, the expression level of VEGF-A increased (compare Control and OIR). The increase in the expression level of VEGF-A did not decrease with the healing of abnormal angiogenesis (compare OIR and ISOX).

[0125] On the other hand, as shown in Figures 3 and 4, when abnormal angiogenesis occurred, the expression levels of the above-mentioned 21 genes (Adm gene, Anxa2 gene, Apln gene, Cd34 gene, Cxcr4 gene, Edn1 gene, Ednra gene, Eng gene, Fn1 gene, Hmox1 gene, Itga1 gene, Loxl2 gene, Mcam gene, Nr4a1 gene, Serpine1 gene, Tnfrsf12a gene, Acta2 gene, Anxa3 gene, Thbs gene, Cdh5 gene, and Esm1 gene) increased. The increase in the expression levels of the 21 genes decreased as abnormal angiogenesis was cured.

[0126] In other words, in the process of abnormal angiogenesis development and the process of abnormal angiogenesis recovery, the expression pattern of VEGF-A did not coincide with that of the above-mentioned 21 genes. This suggests that both (i) a signal transduction system involving VEGF (VEGF-dependent signal transduction system) and (ii) a signal transduction system involving at least one gene selected from the above-mentioned 21 genes (VEGF-independent signal transduction system) are involved in the recovery of abnormal angiogenesis. In other words, this suggests that the therapeutic or preventive agent for abnormal angiogenesis according to one embodiment of the present invention exerts its effect through both the VEGF-dependent signal transduction system and the VEGF-independent signal transduction system. [Industrial Applicability]

[0127] The present invention can be utilized in the treatment or prevention of abnormal angiogenesis.

Claims

1. A therapeutic or preventive agent for abnormal angiogenesis, comprising a histone acetyltransferase inhibitor as an active ingredient.

2. The therapeutic or preventive agent for abnormal angiogenesis according to claim 1, wherein the histone acetyltransferase inhibitor targets one or more substances selected from the group consisting of HAT1, NAA60, GCN5, PCAF, TIP60, MOZ, MORF, HBO1, MOF, TAF1, TIFIIIC90, SRC1, SCR3, P600, CLOCK, NAT10, P300, CBP, and BRD4.

3. The agent for treating or preventing abnormal angiogenesis according to claim 1, wherein the histone acetyltransferase inhibitor is a compound represented by the following chemical formula 1, a derivative thereof, or a salt thereof. 【Chemistry 1】

4. The method of claim 1, wherein the abnormal angiogenesis is the formation of an ischemic area and / or the formation of abnormal blood vessels.

5. The therapeutic or preventive agent for abnormal angiogenesis according to any one of claims 1 to 4, wherein the abnormal angiogenesis is abnormal angiogenesis associated with retinal ischemic disease, cancer, retinopathy of prematurity, age-related macular degeneration, diabetic macular edema, pathological myopia, macular edema associated with retinal vein occlusion, or neovascular glaucoma.

6. Use of a histone acetyltransferase inhibitor for the manufacture of an agent for treating or preventing abnormal angiogenesis.

7. administering a candidate substance for a therapeutic or preventive agent for abnormal angiogenesis to a non-human model organism that has developed abnormal angiogenesis; a measuring step of measuring, in the non-human model organism after the administration step, (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene; A screening method for a therapeutic or preventive agent for abnormal angiogenesis, comprising a determination step of determining whether the candidate substance is a therapeutic or preventive agent for abnormal angiogenesis by comparing the expression level of each gene measured in the measurement step with a control expression level.

8. A method for obtaining data for diagnosing abnormal angiogenesis, comprising a measuring step of measuring, in a biological sample collected from a subject, (i) the expression level of the Ang2 gene, and (ii) the expression level of at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene.

9. A marker for diagnosing abnormal angiogenesis comprising: (i) the Ang2 gene; and (ii) at least one gene selected from the group consisting of the Adm gene, the Anxa2 gene, the Apln gene, the Cd34 gene, the Cxcr4 gene, the Edn1 gene, the Ednra gene, the Eng gene, the Fn1 gene, the Hmox1 gene, the Itga1 gene, the Loxl2 gene, the Mcam gene, the Nr4a1 gene, the Serpine1 gene, the Tnfrsf12a gene, the Acta2 gene, the Anxa3 gene, the Thbs gene, the Cdh5 gene, and the Esm1 gene.

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