JP2002087982A - Vascularization stimulator, vascularization inhibitor and method for screening them - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an excellent vascularization stimulator usable as a medicinal-therapy agent for cardiovascular diseases or injuries, and the like, or for screening vascularization inhibitors. SOLUTION: The vascularization stimulator contains a urotensin II-like peptide compound shown by formula (1) (wherein, R1 is Glu, GlyPro, PyrPro, GlnHisGlyThr, PyrHisGlyThr, GlnHisGlyAla, PyrHisGlyAla, AlaGlyAsn, AlaGly, GlyGly, GlySer or AsnAsn; R2 is Thr, Ala, Leu, Gly, Asn or Phe; R3 is Pro, Ser, Ala or Thr; R4 is Asp or Glu; and R5 is Val or Ile) or its pharmaceutically permissible salt as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an angiogenesis promoting agent, an angiogenesis inhibitor or a screening method thereof useful as a pharmaceutical composition.

[0002]

2. Description of the Related Art Angiogenesis is a phenomenon in which small blood vessels are formed from existing blood vessels. Although the prognosis of various cardiovascular diseases such as myocardial infarction is affected by many factors, the degree of collateral circulation development is considered to be one of the most important prognostic determinants. With sufficient development of collateral circulation,
Even if stenosis or occlusion occurs, ischemia and necrosis of tissue are suppressed, and the size of the infarct is reduced and the prognosis is improved. Conventionally,
Changes in intravascular pressure and blood flow have been considered important as the mechanism of collateral circulation, but cell division with DNA synthesis is observed in vascular endothelial cells and vascular smooth muscle cells during collateral circulation. It is reported that the formation process of collateral circulation is not only a dilation of existing anastomotic blood vessels due to physical factors, but also at least a part of angiogenesis process involving proliferation of vascular wall constituent cells. It is becoming understood. In recent years, attempts have been made to treat cardiovascular diseases with a new treatment called "angiogenesis therapy" (Yanagisawa-Miwa, A. et al., Science, 257, 140).
1, 1992). Angiogenesis therapy is an attempt to actively secure collateral circulation by promoting angiogenesis around ischemic tissue and protect ischemic tissue. It is a cure. However, it has not yet been put to practical use, and development of an excellent drug that can be used for this has been expected. On the other hand, attempts have been made to use substances having angiogenesis promoting activity such as fibroblast growth factor for treating wounds (Ho).
ckel, M. et al., Arch. Surg., 128, 423, 1993). However, this has not yet been put to practical use, and development of excellent drugs is expected.

[0003] On the other hand, there are diseases in which angiogenesis caused by the diseases is related to the progress thereof. Solid tumors, diabetic retinopathy, and inflammatory diseases (rheumatoid arthritis) are
This is a typical example. In order for a solid tumor to grow, it is essential to provide a way to supply nutrients and oxygen and remove waste products by angiogenesis. In addition, in metastasis, which is a major problem in cancer treatment at present, angiogenesis led by cancer cells is an important step from the viewpoint of securing the path. In diabetic retinopathy, the resulting neovascularization itself becomes a disease state and, if left unchecked, leads to blindness. Therefore, suppression of angiogenesis caused by these diseases is considered to lead to prevention and treatment of the progression of the diseases, and search for angiogenesis inhibitors has been vigorously conducted (Folkma
n, J. et al., Science 275, 116, 1996). However, no drug is still clinically satisfactory.

By the way, urotensin II is a cyclic peptide originally isolated from a hormone-secreting organ called urophysisis in teleost fish (Bern, HA et al., J. Endo).
crinol. 45, Xi, 1969). Thereafter, urotensin II having several different chemical structures having a C-terminal cyclic hexapeptide as a common structure has been found in mammals including fish, amphibians and humans (Coulouarn, Y.
et al., Proc. Natl. Acad. Sci. 95: 15803, 1998, Cou.
louarn, Y., et al., FEBS Letters 457: 28-32, 1999
And Hoare, SRJ Trends Pharmacol. Sci. 21: 80,
2000). These urotensin II actions include vasoconstrictor action (Itoh, H., et al., Eur. J. Pharmacol. 149:
61-66, 1988 and Ames, RS et al., Nature 401: 28
2, 1999) and a decrease in cardiac contractility (Ames, RS et a
l., Nature 401: 282, 1999).

[0005] Urotensin II also acts via a urotensin II receptor on the cell membrane (Itoh,
H., et al., Eur. J. Pharmacol. 149: 61-66, 198
8). Urotensin II, as one of the manifestations of its effects, is GPR14 or SENR (sensory epithelium ne
A known G protein-coupled receptor called uropeptide-like receptor (Marchese, A., et al., Genomics 2
9: 335, 1995 and Tal, M., et al., Biochem. Biophys.
Res. Commun. 209: 752, 1995) is known to exert an effect by stimulating, that is, urotensin II is known to be an endogenous substance that stimulates G protein-coupled receptor GPR14. (Nothacker, H.-P., et al., N
ature Cell Med 1: 383, 1999, Liu, Q., et al., Bioc
hem. Biophys. Res. Commun. 266: 174, 1999, Mori,
M., et al., Biochem. Biophys. Res.Commun. 265: 12
3, 1999 and Ames, RS et al., Nature 401: 282, 1
999). This urotensin II is first produced in vivo by a precursor, preprourotensin II, and then produced by the action of peptidases such as prohormone convertase (Coulouran, Y., et al., Proc. Natl. Acad. Sc.
i. 95: 15803, 1998).

[0006]

However, pharmacotherapy for cardiovascular diseases or wounds utilizing the angiogenesis promoting action has not yet been put to practical use, and an excellent angiogenesis promoting agent which can be used therefor has not been developed. Provision is an important issue. In addition, there is still no clinically satisfactory angiogenesis inhibitor, and there is no way to screen for such an excellent angiogenesis inhibitor, so solid tumors involving angiogenesis, diabetic retinopathy, and inflammation There is no sufficient treatment method for sexual diseases and the like, and it is particularly important to provide a clinically satisfactory new angiogenesis inhibitor and a screening method thereof.

[0007]

The present inventors have conducted intensive studies to find a compound that promotes angiogenesis or a compound that inhibits angiogenesis, and as a result, found that a urotensin II-like peptide compound or a pharmacologically acceptable urotensin II-like peptide compound. Have been found to be extremely useful as angiogenesis promoters. Furthermore,
A method for screening an angiogenesis inhibitor using an angiogenesis effect of a urotensin II-like peptide compound or a pharmacologically acceptable salt thereof as an index, and further antagonizing the angiogenesis effect of a urotensin II-like peptide compound It has been found that a compound or a pharmacologically acceptable salt thereof is extremely useful as an angiogenesis inhibitor, which has led to the present invention. That is, the present invention relates to the following (I) to (XII).

(I) An angiogenesis-promoting agent comprising, as an active ingredient, a urotensin II-like peptide compound or a pharmacologically acceptable salt thereof. (II) The urotensin II-like peptide compound has the general formula (1)

[0009]

Embedded image (Wherein, R ₁ is Glu-, Gly-Pro-, Pyr-Pro-, Gln-His-
Gly-Thr-, Pyr-His-Gly-Thr-, Gln-His-Gly-Ala-, Pyr-
His-Gly-Ala-, Ala-Gly-Asn-, Ala-Gly-, Gly-Gly-, Gl
y-Ser- or Asn-Asn-, wherein R ₂ is Thr, Ala, Le
u, Gly, Asn or Phe; R ₃ is Pro, Ser, Ala
Or Thr, R ₄ represents Asp or Glu, and R _4
5 represents -Val or -Ile. (I) the angiogenesis promoting agent according to (I). (III) The angiogenesis promoting agent according to (I), wherein the urotensin II-like peptide compound has the following amino acid sequence.

[0010]

Embedded image (IV) The angiogenesis promoting agent according to (I), wherein the urotensin II-like peptide compound has the following amino acid sequence.

[0011]

Embedded image (V) The angiogenesis promoting agent according to (I), wherein the urotensin II-like peptide compound has the following amino acid sequence.

[0012]

Embedded image (VI) An angiogenesis inhibitor is screened using the peptide compound or salt thereof according to (I) as an indicator of angiogenesis.
A method comprising: (VII) The method according to (VI), wherein the peptide compound or a salt thereof is as described in (II). (VIII) The method according to (VI), wherein the peptide compound or a salt thereof is as described in (III). (XI) The method according to (VI), wherein the peptide compound or a salt thereof is as described in (IV). (X) The method according to (VI), wherein the peptide compound or a salt thereof is one described in (V). (XI) The method according to any one of (VI) to (X), which utilizes a reaction in the cornea of an experimental animal. (XII) An angiogenesis inhibitor comprising, as an active ingredient, a compound or a pharmacologically acceptable salt thereof which antagonizes the angiogenesis effect of a urotensin II-like peptide compound.

[0013]

DETAILED DESCRIPTION OF THE INVENTION In the present invention, urotensin II
-Like peptide compounds have the following partial structure

[0014]

Embedded image Such as urotensin II derived from a living body and a peptide compound having a structure similar thereto, specifically, for example, a peptide compound represented by the following general formula (1).

[0015]

Embedded image (Wherein, R ₁ is Glu-, Gly-Pro-, Pyr-Pro-, Gln-H
is-Gly-Thr-, Pyr-His-Gly-Thr-, Gln-His-Gly-Ala-
, Pyr-His-Gly-Ala-, Ala-Gly-Asn-, Ala-Gly-, Gly
-Gly-, Gly-Ser- or Asn-Asn-, and R ₂ represents Th
represents r, Ala, Leu, Gly, Asn or Phe, and R ₃ is Pr
o, Ser, Ala or Thr, and R ₄ is Asp or Glu
Are shown, _{R 5} represents a -Val or -Ile. )

In the general formula (1), R ₁ , R ₂ , R ₃ , R ₄ and R ₅ , various amino acids constituting the amino acid are N-terminal on the left side and C-terminal on the right side.

[0017] Formula _{(1), R 1, R} 2, R 3, for _{R 4} and _{R 5,} glutamic acid and Glu, glycine and Gly, proline and Pro, pyroglutamic acid and Pyr, glutamine and Gln , His is histidine, Thr is threonine, Ala is alanine, As
n is asparagine, Ser is serine, Leu is leucine, Phe is phenylalanine, Val is valine, Lys is lysine, Cys is cysteine, Asp
Is aspartic acid, Trp is tryptophan, Il
e shows isoleucine.

Specific examples of the peptide compound represented by the general formula (1) are, for example, the compounds shown in Table 1 below.

[0019] Table 1 Examples Compound of the peptide compound represented by the general formula _{(1) R 1 R 2 R} 3 R 4 R 5 1 Glu- Thr Pro Asp -Val 2 Gly-Pro- Thr Ser Glu -Val 3 Pyr- His-Gly-Thr-Ala Pro Glu -Ile 4 Gln-His-Gly-Thr-Ala Pro Glu -Ile 5 Gln-His-Gly-Ala-Ala Pro Glu -Ile 6 Pyr-His-Gly-Ala-Ala Pro Glu -Ile 7 Ala-Gly-Asn-Leu Ser Glu -Val 8 Ala-Gly- Thr Ala Asp -Val 9 Gly-Gly-Gly Ala Asp -Val 10 Gly-Gly-Gly Ala Asp -Ile 11 Gly-Gly- Asn Thr Glu -Val 12 Gly-Ser- Asn Thr Glu -Val 13 Gly-Gly- Asn Ser Glu -Val 14 Gly-Ser- Gly Ala Asp -Val 15 Ala-Gly- Thr Thr Glu -Val 16 Gly-Ser- Thr Ser Glu -Val 17 Asn-Asn- Phe Ser Glu -Val

In the general formula (1), R ₁ , R ₂ , R ₃ , R ₄ and R ₅ , the various amino acids constituting them are L
Body, D-form or DL-form, or L-form, D-form
It may be any of those in which a body or a DL body is appropriately mixed, but those in which all are L bodies are preferable.

The urotensin II-like peptide compound used in the present invention can be synthesized, for example, by extraction from a living body, or by using a peptide synthesizer reported by Mori, M. et al. (Biochem. Biophys. Res. Com.
mun. 265: 123, 1999).

Urotensin II used in the present invention
Is a known peptide compound derived from, for example, mammals including humans, amphibians, or fish, and includes, for example, humans, pigs, rats, mice, frogs, goby, carp, trout, and suck.
er, flounder, sturgeon, paddlefish, skate, petorom
and urotensin II, specifically, compounds 1 (human) and compound 2 shown in Table 1 above.
(Pig), compounds 3, 4 (rat), compounds 5, 6 (mouse), compound 7 (frog), compound 8 (goby), compounds 9-12 (carp), compound 13 (mass), compound 14
(Sucker), compound 15 (flounder), compound 16 (st
urgeon or paddlefish) or compound 17 (skate, petor)
omyzon, dogfish and lamprey).

Specific examples of the pharmacologically acceptable salts used in the present invention include, for example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, and hydrogen fluoride. Acid halides such as acid salts, hydrochlorides, hydrobromides or hydroiodides, acid addition salts with mineral acids such as sulfates, nitrates, sulfonic acids or phosphates, methanesulfonates Or a lower alkyl sulfonate such as ethane sulfonate, a halogeno lower alkyl sulfonate such as trifluoromethane sulfonate, an unsubstituted aryl sulfonate such as benzene sulfonate, and a substituted aryl such as p-toluene sulfonate. Sulfonate, acetate, lactate, fumarate, succinate, citrate, tartrate, oxalate, benzoate, Rain salts, organic acid salts or glutamate or amino acid salts such as aspartate salts and malic acid or phthalic acid salt can be exemplified.

When the urotensin II-like peptide compound used in the present invention or a pharmacologically acceptable salt thereof is used as an angiogenesis promoting agent, various injections, percutaneous mucosal administration agents, oral agents, etc. Dosage forms are possible and are used in various dosage forms depending on the dosage form. Examples of the mode of administration include direct injection into the heart muscle from the intraventricular cavity using a catheter, and local release or application using a catheter to a stenosis or occlusion in a coronary artery. Dosage forms include, for example, injections for intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, etc., suspension injections, dissolution-type injections such as freeze-drying, suppositories, nasal preparations, etc. Transmucosal agent,
Eye drops, inhalants, patches, ointments or sprays,
Or oral preparations such as tablets, capsules, granules, fine granules, powders and suspensions can be mentioned. The dose varies depending on the patient's condition, age, body weight, therapeutic purpose, administration method, administration timing, administration days or administration period, but is generally 0.001 to 20 mg / day.

For formulating the angiogenesis promoting agent of the present invention, generally known methods are applied. For example, when the pharmaceutical carrier is a liquid, it is dissolved or dispersed, and when the pharmaceutical carrier is a powder, it is mixed or adsorbed on the powder, and depending on the purpose, these are pharmaceutically acceptable. Preservatives, stabilizers, antioxidants, excipients,
Binders, disintegrants, wetting agents, lubricants, coloring agents, fragrances, corrigents, skins, suspending agents, emulsifiers, solubilizing agents, buffers,
It may also contain tonicity agents, plasticizers, surfactants or soothing agents.

The pharmaceutical carrier used in the angiogenesis promoting agent of the present invention includes, for example, water, distilled water for injection, physiological saline,
Purified water, glucose, fructose, sucrose, lactose (lactose), starch, cellulose, methylcellulose,
Carboxymethylcellulose, hydroxypropylcellulose, glycerin, inositol, mannitol, xylitol, sorbitol, glucuronic acid, hyaluronic acid, heparin, chitin, chitosan, glycine, alanine, phenylalanine, proline, serine, cysteine, aspartic acid, glutamic acid, lysine, arginine, Human serum albumin, human serum globulin, collagen, gelatin, alginic acid, talc, sodium citrate, calcium carbonate, calcium hydrogen phosphate, magnesium stearate, urea, silicone resin, sorbitan fatty acid ester, glycerin fatty acid ester, ascorbic acid, alpha tocopherol , Ethylene glycol, polyethylene glycol or silicon dioxide.

The content of the peptide compound in the angiogenesis-promoting agent of the present invention varies depending on the preparation.
00% by weight. Also, for example, in the case of an injection solution,
Usually, it is preferable to contain 0.1 to 5% by weight of the active ingredient.

As a method for screening an angiogenesis inhibitor using the urotensin II-like peptide compound of the present invention or a salt thereof as an indicator, for example, the cornea of an experimental animal (eg, an animal such as a rabbit, rat or mouse) may be used. And the method utilizing the reaction in the above. As an example of the practice of this method, the cornea of the animal was injured with a scalpel and two pockets were made to insert the pellet into the vascular plexus near the top of the eyeball or on the opposite side.
The tip of the pocket was 2 mm away from the limbus (the junction between the cornea and the sclera). The test compound was dissolved or suspended in the pocket, allowed to stand at a negative temperature overnight, and a solidified pellet or a control pellet was inserted first, and then a pellet containing human urotensin II was inserted. After a certain period of time (for example, 5 days), the length of a blood vessel newly formed from the lymph bath toward the pellet was measured using a measure, and the measured value was used as an index of inhibition of angiogenesis of the test compound.

Another method for screening an angiogenesis inhibitor using the angiogenesis effect of the urotensin II-like peptide compound of the present invention or a salt thereof as an index includes, for example,
Itoh, H. et al. Reported (Eur. J. Pharmacol. 149: 61-
66, 1988) by utilizing the action of urotensin II via the urotensin II receptor on the cell membrane to perform a contractile reaction on isolated rat thoracic aorta specimens to inhibit angiogenesis. Binding assay using gooseurotensin II labeled with isotope and membrane fraction of rat thoracic aorta (Ito
h, H., et al., Eur. J. Pharmacol. 149: 61-66, 198
8), a method of screening an angiogenesis inhibitor, the method of Liu, Q., et al. (Biochem. Biophys.
s. Commun. 266: 174, 1999), using HEK293 cells expressing a large number of rat GPR14, utilizing the fact that urotensin II is an endogenous substance that stimulates the G protein-coupled receptor GPR14. A method for screening, a method for screening an angiogenesis inhibitor by immunohistological staining of the spinal cord of rats using a commercially available antibody against rat urotensin II, or a method for inhibiting prohormone convertase (J. Biol. Chem. 273:
26589-26595, 1998), and a method for screening an angiogenesis inhibitor.

The compound that antagonizes the angiogenic action of the urotensin II-like peptide compound or a pharmacologically acceptable salt thereof includes, for example, a compound that inhibits the angiogenic action of the urotensin II-like peptide compound.

When a compound or a pharmacologically acceptable salt thereof which antagonizes the angiogenic action of the urotensin II-like peptide compound is used as an angiogenesis inhibitor, it is administered orally or parenterally. . The dose varies depending on the condition, age, body weight, treatment purpose or administration method of the patient, but is generally 1 to 600 mg / day. Examples of the form of the preparation include tablets, granules, fine granules, powders, capsules, inhalants, patches, eye drops, suppositories, sprays, ointments and injections.

The angiogenesis inhibitor of the present invention is usually formulated by mixing the active ingredient with a carrier, or diluting with a carrier, or containing the active ingredient in a carrier. It may be in the form of paper or other containers. When the carrier acts as a diluent, it may be a solid, semi-solid or liquid material, a vehicle for the active ingredient,
Acts as an excipient or vehicle. That is, the formulation comprises tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsifiers, solutions, syrups or aerosols (as a solid or liquid medium), for example, one active ingredient at a time.
Ointments, soft gelatin capsules, hard gelatin capsules, suppositories, sterile injectable solutions or sterile packaged powders containing 0% by weight may be employed.

Some examples of suitable carriers, excipients or diluents for use in the angiogenesis inhibitors of the present invention include lactose (lactose), dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, Alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline, syrup, methylcellulose,
It may include -methyl hydroxybenzoate, -propyl hydroxybenzoate, talc, magnesium stearate or mineral oil. The formulation may additionally contain lubricants, wetting agents, emulsifying agents, suspending agents, preservatives, sweetening agents or flavoring agents. Additionally, the compositions can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient using methods known to those skilled in the art.

[0034]

EXAMPLES Examples of the present invention are described below.
Test examples and preparation examples using II are shown, but the present invention is not limited to these.

Example 1 Angiogenesis-Promoting Action of Urotensin II The angiogenesis-promoting action of human urotensin II was determined by the rabbit corneal method (Abramovitch, R., et al., FEBS Letters 425, 441,
1998).

Experimental method Female New Zealand white rabbit (3.4-3.7 k
g, Oriental yeast) was anesthetized by intravenous administration of sodium pentobarbital (25 mg / kg). Furthermore, local anesthesia was performed with oxybuprocaine hydrochloride. A pocket for inserting a pellet was cut in the center of the cornea with a scalpel, an ophthalmic spatula was inserted into the incision, and two places were created toward the parietal plexus of the eyeball or toward the opposite plexus. . The tip of the pocket was 2 mm away from the limbus (the junction between the cornea and the sclera). A human urotensin II or empty pellet was inserted into the pocket using an ophthalmic spatula. As a pellet, a 10% ethylene vinyl acetate solution itself dissolved in dichloromethane as a control,
The solution was prepared by dissolving 10 μg of human urotensin II in the solution, further diluting it 200 times with a 10% ethylene vinyl acetate solution, and leaving the above solution at −20 ° C. overnight to remove the solvent. A pellet (about 2 mm in diameter) was used. After inserting the pellet, 10m of ampicillin sodium
g / ml was instilled and 14 mg / kg was administered intravenously. 5 and 7 days after the insertion of the pellet, the length of the newly formed blood vessel from the limbus to the pellet under sodium pentobarbital anesthesia was measured using a measure,
It was used as an index of angiogenesis.

Results The results are shown in Table 2.

Table 2. Promoting angiogenesis by urotensin II in rabbit cornea {Length of newly formed blood vessel (in mm) is indicated} Contents of pellet to be inserted into cornea 5 days after insertion of pellet 7 days after insertion of pellet Control (nothing Insert a pellet that does not contain) 0.0 0.0 Urotensin II 0.05 μg 0.4 0.7 Urotensin II 10 μg 1.5 2.1

As is evident from Table 2, human urotensin II exhibited a pro-angiogenic effect in a dose-dependent manner. The effect was recognized as early as 5 days after the insertion of the pellet, and the effect was extremely strong, indicating the usefulness of the angiogenesis promoter of the present invention. In addition, no side effect or toxicity that caused inflammation such as clouding of the cornea or accumulation of leukocytes due to human urotensin II was observed.

Example 2 Method for Screening an Angiogenesis Inhibitor Using Reaction in Rabbit Cornea Using urotensin II, a method for finding an angiogenesis inhibitor using a reaction in rabbit cornea was examined.

Experimental Method A female New Zealand white rabbit (3-4 kg, Oriental yeast) was used in sodium pentobarbital (2
General anesthesia was performed by intravenous administration of 5 mg / kg). Furthermore, local anesthesia was performed with oxybuprocaine hydrochloride.
In the pocket for inserting the pellet, scratch the center of the cornea with a scalpel, insert an ophthalmic spatula into the cut,
Two sites were created toward the vascular plexus near the parietal of the eyeball or on the opposite side. The tip of the pocket was 2 mm away from the limbus (the junction between the cornea and the sclera). The test compound was placed in a pocket using an ophthalmic spatula.
% Ethylene vinyl acetate solution or suspended and solidified at −20 ° C. overnight and solidified (diameter approx.
mm) or a control pellet is inserted first, followed by a pellet (about 1 mm in diameter) containing 10 μg of human urotensin II.
mm) was inserted. For protection against infection, 10 mg / ml of ampicillin sodium was instilled, and 14 mg / kg
Was administered intravenously. Five days after the insertion of the pellet, the length of the blood vessel newly formed from the lymph bath toward the pellet under sodium pentobarbital anesthesia was measured using a measure, and used as an index of the angiogenesis inhibition of the test compound.

Example 3 The toxicity of the angiogenesis promoter of the present invention was examined using rats.

Toxicity test Male Wistar rats (250 g, Charles River Japan) were anesthetized with pentobarbital, and rat urotensin II (compound 3 in Table 1 above) was treated with 10, 30, 10 rats.
0, 300, and 1000 pmol / kg were continuously administered intravenously to examine toxic effects. Neither toxic effects on blood pressure, heart rate or electrocardiogram nor any other toxic effects were observed, confirming the high safety of the angiogenesis promoting agent of the present invention.

Formulation Example 1 Purified water is added to 30 parts by weight of human urotensin II to make the total amount 2000 parts by weight, dissolved, and then sterilized and filtered using a Millipore filter GS type. This filtrate 2
mg was placed in a 10 ml vial and freeze-dried to obtain a freeze-dried injection containing 30 μg of human urotensin II in one vial.

Formulation Example 2 Purified water was added to 30 parts by weight of rat urotensin II (compound 3 in Table 1 above) to make the total amount 2000 parts by weight, dissolved, and then sterilized and filtered using a Millipore filter GS type. I do. 2 mg of the filtrate was placed in a 10 ml vial and freeze-dried to obtain a freeze-dried injection containing 30 μg rat urotensin II in one vial.

Formulation Example 3 Rat urotensin II (compound 3 in Table 1 above) was added to 100
Parts by weight, horse starch 30 parts by weight, crystalline lactose 15
0 parts by weight, crystalline cellulose 108 parts by weight and magnesium stearate 2 parts by weight were mixed with a V-type mixer, and one tablet 6
The tablets were compressed at 0 mg to give tablets containing 20 mg of the peptide compound per tablet.

Formulation Example 4 To 50 mg of human urotensin II, 400 mg of microcrystalline cellulose, 10 mg of silicon dioxide and 5 mg of magnesium stearate were added to obtain a hard gelatin capsule.

Formulation Example 5 Rat urotensin II (compound 3 in Table 1 above) was treated with 50 m
To 400 g of microcrystalline cellulose, 10 mg of silicon dioxide and 5 mg of magnesium stearate were added to obtain a hard gelatin capsule.

[0049]

Industrial Applicability The urotensin II-like peptide compound of the present invention or a pharmacologically acceptable salt thereof has an excellent angiogenesis promoting action and is useful as a prophylactic or therapeutic drug based on this action. Specifically, it is useful as a prophylactic and therapeutic agent (angiogenesis promoting agent) used in angiogenesis therapy.
More specifically, circulatory diseases, for example, acute myocardial infarction,
Unstable angina, chronic stable angina, old myocardial infarction, thromboembolism due to atrial fibrillation, generalized intravascular coagulation, graft occlusion after coronary artery bypass grafting, percutaneous artery Stenosis and occlusion of coronary arteries after plastic surgery, thrombotic complications after artificial heart valve replacement, pulmonary thrombosis / infarction, cerebrovascular accident (transient cerebral ischemic attack, cerebral infarction), peripheral arterial occlusion (obstruction) Atherosclerosis, obstructive thromboangiitis, obstruction after revascularization), glomerulonephritis, nephrotic syndrome, thrombosis of essential thrombosis and thrombotic thrombocytopenic purpura, hemolytic uremic syndrome, antiphosphorus Prevention and treatment of lipid-antibody syndrome, Kawasaki disease, eclampsia, and Behcet's disease; Wound and gastrointestinal tissue damage after various operations such as skin transplantation It is effective for the treatment etc.). In addition, a method for screening an angiogenesis inhibitor using a urotensin II-like peptide compound or a pharmacologically acceptable salt thereof using the angiogenesis action as an index was found. This method is useful as a method for finding an angiogenesis inhibitor. Further, the compound or a pharmacologically acceptable salt thereof which antagonizes the angiogenic action of the urotensin II-like peptide compound shows a strong angiogenesis inhibitory action in the following disease states, and a prophylactic or therapeutic agent based on this action. Useful as Specifically, tumors such as malignant melanoma, malignant lymphoma, gastric cancer, intestinal cancer, lung cancer, pancreatic cancer, esophageal cancer, breast cancer, liver cancer, ovarian cancer, uterine cancer, prostate cancer, kidney cancer, bladder cancer, brain tumor, Prophylactic and therapeutic agents for Kaposi's sarcoma, hemangiomas, osteosarcomas, sarcomas, and hemangiofibromas, eye diseases such as diabetic retinopathy, retinopathy of prematurity, sickle cell retinopathy, retinal vein occlusion, corneal transplantation Or angiogenesis with cataract surgery, angiogenic glaucoma, iris rubeosis, and preventive and therapeutic agents for angiogenesis in the cornea by long-term wearing of contact lenses,
It is also useful as a prophylactic and therapeutic agent for inflammatory diseases such as rheumatoid arthritis and psoriasis.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A61P 13/00 A61P 17/02 17/02 19/02 19/02 27/02 27/02 29/00 29 / 00 35/00 35/00 43/00 107 43/00 107 A61K 37/02

Claims (12)

[Claims] 1. An angiogenesis promoting agent comprising a urotensin II-like peptide compound or a pharmacologically acceptable salt thereof as an active ingredient. 2. A urotensin II-like peptide compound represented by the general formula (1): (Wherein, R ₁ is Glu-, Gly-Pro-, Pyr-Pro-, Gln-H
is-Gly-Thr-, Pyr-His-Gly-Thr-, Gln-His-Gly-Ala-
, Pyr-His-Gly-Ala-, Ala-Gly-Asn-, Ala-Gly-, Gly
-Gly-, Gly-Ser- or Asn-Asn-, and R ₂ represents Th
represents r, Ala, Leu, Gly, Asn or Phe, and R ₃ is Pr
o, Ser, Ala or Thr, and R ₄ is Asp or Glu
Are shown, _{R 5} represents a -Val or -Ile. )). 3. The angiogenesis promoting agent according to claim 1, wherein the urotensin II-like peptide compound has the following amino acid sequence. Embedded image 4. The angiogenesis promoting agent according to claim 1, wherein the urotensin II-like peptide compound is a compound having the following amino acid sequence. Embedded image 5. The angiogenesis promoting agent according to claim 1, wherein the urotensin II-like peptide compound is a compound having the following amino acid sequence. Embedded image 6. A method comprising screening an angiogenesis inhibitor using the angiogenic action of the peptide compound or a salt thereof according to claim 1 as an index. 7. The method according to claim 6, wherein the peptide compound or a salt thereof is one according to claim 2. 8. The method according to claim 6, wherein the peptide compound or a salt thereof is the one according to claim 3. 9. The method according to claim 6, wherein the peptide compound or a salt thereof is as defined in claim 4. 10. The method according to claim 6, wherein the peptide compound or a salt thereof is as described in claim 5. 11. The method according to claim 6, wherein a reaction in a cornea of an experimental animal is utilized. 12. An angiogenesis inhibitor comprising, as an active ingredient, a compound or a pharmacologically acceptable salt thereof which antagonizes the angiogenesis effect of a urotensin II-like peptide compound.