JP2003300902A - Angiogenesis inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic agent for diseases in which neovascularization involves as a result of researches concerning influence of Suc-Val-Pro-HNCH(CH<SB>2</SB>Ph)P(O)(OPh)<SB>2</SB>having a chymase-inhibiting action on neovascularization. <P>SOLUTION: The Suc-Val-Pro-HNCH(CH<SB>2</SB>Ph)P(O)(OPh)<SB>2</SB>having the chymase- inhibiting action exhibits neovascularization-inhibiting action and is expected as a prophylactic or therapeutic agent for diseases in which neovascularization involves, particularly, intraocular neovascular diseases such as diabetic retinopathy, maculopathy, retinal vein blockage, premature infant retinitis and neovascular glaucoma. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an angiogenesis inhibitor containing Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ having a chymase inhibitory activity as an active ingredient.

[0002]

2. Description of the Related Art Angiogenesis is a phenomenon in which a new blood vessel network, that is, a new blood vessel is formed from existing blood vessels. Angiogenesis is found in various physiological and pathological conditions in the body. Physiological angiogenesis is recognized during luteinization and placenta formation, and angiogenesis in a pathological state is recognized by inflammation, wound healing, tumor growth and the like.

[0003] The most important characteristic of the tissue construction of the eyeball is the transparency of the tissue, and once angiogenesis is induced, the transparency is impaired and a remarkable visual dysfunction occurs. In the field of ophthalmology, various diseases such as diabetic retinopathy, age-related macular degeneration, and retinopathy of prematurity are considered to fall into the category of angiogenic diseases and are called intraocular angiogenic diseases (ophthalmology,
40 , 1677-1683 (1998)).

Examples of angiogenic diseases include intraocular angiogenic diseases, solid tumors, rheumatoid arthritis, psoriasis, angiogenesis in atheromatous lesions, ischemic heart disease and arteries. Examples include collateral circulation associated with sclerosis, hemangiomas, and Kaposi's sarcoma, but research on solid tumors is the most advanced (Latest Medicine, 53 ,
2671-2678 (1998)). It is also well known that angiogenesis occurs during the healing process of wounds.

On the other hand, chymase is present in tissues of the whole body such as the digestive tract, skin, lungs, joints (synovium) as well as tissues of the cardiovascular system, and is affected by cardiovascular lesions, inflammation, immune function, tissue structure modification, etc. It has been reported that it is involved in the exertion of physiological functions of A. (medical history, 174 , 743-747 (1995)). Also,
Chymase causes arteriosclerosis, myocardial infarction, heart failure, vascular restenosis after percutaneous coronary angioplasty (PTCA) (vascular and endothelium, 5 , 461-468 (1995)), diabetic complications (Biol. Ch.
em. Hoppe-Seyler, 369 , Suppl. 299-305 (1988)),
Allergic diseases (J. Biochem., 103 , 820-822 (1988)
), Asthma (J. Pharmacol. Exp. Ther., 244 , 133-137
(1988)), Rheumatoid arthritis (Ann. Rheum. Dis., 56 ,
151-161 (1997)).

However, no report has yet been made on the relationship between chymase and angiogenesis, particularly intraocular neovascularization.

[0007]

As described above, the relationship between chymase and angiogenesis has not yet been elucidated in detail, and research on the effect of compounds having a chymase inhibitory effect on angiogenesis is extremely difficult. This is an interesting subject. Further, as research on therapeutic agents for diseases associated with angiogenesis,
There have been many studies on drugs for the treatment of cancer, etc., but there are few studies on the drugs for the treatment of intraocular angiogenic diseases, and research and development of drugs in this field are desired. .

[0008]

The inventors of the present invention have conducted extensive studies on the relationship between angiogenesis and compounds having various chemical structures having a chymase inhibitory action.
It was found that Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ inhibits angiogenesis. That is, Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ having a chymase inhibitory action shows an angiogenesis inhibitory action, and diseases associated with angiogenesis, particularly diabetic retinopathy, macular degeneration , Retinal vein occlusion, retinopathy of prematurity,
It has been found that it is expected as a prophylactic or therapeutic agent for intraocular angiogenic diseases such as neovascular glaucoma.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention uses Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ (hereinafter referred to as the present compound unless otherwise specified) having an inhibitory effect on chymase as an active ingredient. The present invention relates to an angiogenesis inhibitor.

This compound is a compound of Biochemistry, 30 , 485-493.
It is a known peptide derivative described in (1991).

As will be described in detail in the section of the pharmacological test described below,
Chicken chorioallantoic membrane (chorioallant), which is one of the widely used assay systems for evaluating angiogenesis in vivo
As a result of examination using the oic membrane (CAM) method, it was found that this compound has an inhibitory effect on angiogenesis.

The present invention provides a preventive or therapeutic agent for diseases associated with angiogenesis, and examples of the diseases include intraocular angiogenic diseases, solid cancer, rheumatoid arthritis and psoriasis. ), Angiogenesis in atheromatous lesions, collateral blood vessel formation associated with ischemic heart disease and arteriosclerosis, hemangiomas, Kaposi's sarcoma and the like. by the way,
As research on therapeutic agents for diseases associated with angiogenesis, many researches on drugs for treating cancer etc. have been conducted, but research on drugs for intraocular angiogenic diseases has been conducted. At the very least, the main object of the present invention is to provide a drug useful for this intraocular angiogenic disease. Of course, the present invention is widely applicable to diseases associated with angiogenesis, and the subject is not limited to intraocular angiogenic diseases.

[0013] In intraocular angiogenic diseases, angiogenesis itself can be a determinant that influences the disease state, and therefore inhibition of angiogenesis enables prevention and treatment of these diseases (inflammation and immunity, 4 , 343). -349 (1996)), the results of this test show that this compound, which has a chymase inhibitory effect, is associated with angiogenesis-related diseases, in particular diabetic retinopathy, macular degeneration, retinal vein occlusion, retinopathy of prematurity, and blood vessels. It is shown to be useful as a prophylactic or therapeutic agent for intraocular angiogenic diseases such as neonatal glaucoma.

Intraocular neovascularization is classified by site into retinal neovascularization, intravitreal neovascularization, iris neovascularization, and choroidal neovascularization.

Examples of intraocular neovascular diseases associated with retinal neovascularization and intravitreal neovascularization include intraocular neovascular diseases such as diabetic retinopathy, central retinal vein occlusion and retinopathy of prematurity.

As an intraocular neovascularization disease accompanied by iris neovascularization, intraocular neovascularization such as central retinal vein occlusion, diabetic retinopathy, central retinal artery occlusion, carotid artery disease, uveitis, intraocular tumor, etc. Diseases are mentioned.

As an intraocular neovascular disease accompanied by choroidal neovascularization, age-related macular degeneration (senile discoid macular degeneration),
Retinitis pigmentosa, severe myopia, ocular histoplasmosis (pres
umedocular histoplasmosis (POHS), choroidal rupture, sudden choroidal neovascularization and other intraocular neovascular diseases.

The compound can be administered orally or parenterally. Examples of dosage forms include tablets, capsules, granules, powders, injections and eye drops. These can be prepared using a commonly used technique. For example, in the case of oral preparations such as tablets, capsules, granules and powders, bulking agents such as lactose, crystalline cellulose, starch and vegetable oils; lubricants such as magnesium stearate and talc; hydroxypropyl cellulose, polyvinylpyrrolidone etc. A binder such as carboxymethylcellulose calcium, a low-substituted hydroxypropylmethylcellulose or the like, a disintegrator such as hydroxypropylmethylcellulose, macrogol, a silicone resin or the like, a film forming agent such as a gelatin film, etc. It may be formulated. For eye drops, sodium chloride,
Isotonic agents such as concentrated glycerin; buffering agents such as sodium phosphate and sodium acetate; surfactants such as polyoxyethylene sorbitan monooleate, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil; sodium citrate, edet The above compound can be formulated by using a stabilizer such as sodium acid salt; a preservative such as benzalkonium chloride and paraben, if necessary. The pH may be in the range acceptable for ophthalmic preparations, but is preferably in the range of 4-8. If it is an eye ointment, white petrolatum,
It can be prepared using a commonly used base such as liquid paraffin.

The dose can be appropriately selected according to the symptoms, age, dosage form, etc., but in the case of eye drops, 0.0001 to 5% (w
/ V), preferably 0.001 to 3% (w / v), and particularly preferably 0.001 to 1% (w / v) per day.
~ It may be applied several times. If it is an oral preparation, it is usually 0.1 to 5000 mg, preferably 1 to 1000 m per day.
g may be administered once or in several divided doses.

Formulation examples and results of pharmacological tests are shown below, but these examples are for the purpose of better understanding of the present invention, and do not limit the scope of the present invention.

[0021]

EXAMPLES [Formulation Example] The following are general formulation examples of the oral preparation, eye drop preparation and injection preparation of the present invention.

[0022] 1) Capsules   Prescription (in 150 mg)     This compound 5.0 mg     Lactose 145.0 mg

By changing the mixing ratio of the present compound and lactose, the amount of the present compound was 10.0 mg / capsule, 3
0.0 mg / capsule, 50.0 mg / capsule, 10
Capsules of 0.0 mg / capsule can also be prepared.

[0024] 2) Eye drops (in 10 ml)     This compound 1 mg     Concentrated glycerin 250mg     Polysorbate 80 200 mg     Sodium dihydrogen phosphate dihydrate Suitable amount     1N sodium hydroxide suitable amount     1N hydrochloric acid suitable amount     Sterile purified water

By appropriately changing the amounts of the present compound and additives, the concentration of the present compound is 0.0001%, 0.001%.
%, 0.005%, 0.05%, 0.1%, 0.5%,
Eye drops of 1.0%, 3.0%, 5.0% (w / v) can also be prepared.

[0026] 3) Injection (in 100 ml)     The present compound 10.0 to 100.0 mg     1N sodium hydroxide suitable amount     1N hydrochloric acid suitable amount     Saline solution

[Pharmacological test] 1. Angiogenesis inhibitory effect by chorioallantoic membrane method As one of the in vivo angiogenesis evaluation models that are widely used to measure the inhibitory effect of drugs on angiogenesis, the chicken chorioallantoic membrane (CAM) method using a fertilized egg embryo ( Biochem. Bio
phys. Res. Commun., 174 , 1070-1076 (1991)). Therefore, the inhibitory effect of the present compound on angiogenesis was examined with reference to the method described in the above literature.

(Preparation of pellets for administration) 1. Methylcellulose 4000 cP (0.1 g) is dissolved in sterile purified water (10 ml) to give a 1% methylcellulose solution.

2. This compound is dissolved in ethanol / sterilized purified water (1/1) to prepare a 24 μmol / ml compound solution.

3. 24 μmol / ml of this compound solution (0.20 ml) and 1% methylcellulose solution (0.2
0 ml) and mixed with the present compound mixture (12 μmol / m
l).

4. The compound mixture (10 μl) is air-dried on a paraffin film having a diameter of 3 mm to obtain the compound pellet (0.12 μmol / pellet).

(Experimental Method) 1. Immediately after the arrival of hatched eggs (white leghorn) 3 days after fertilization, the surface of the eggshell was washed and disinfected with 70% ethanol. These hatched eggs were placed in a 6-well culture plate with the air phase facing upwards, and 3 eggs were placed in each, and then the CO ₂ incubator (37
° C., 95% humidity and incubated for 20 minutes at 5% CO ₂₎ in the.

2. The hatched eggs were taken out from the CO ₂ incubator, and about 1 cm square holes were opened in the eggshell above the air phase of the hatched eggs in a clean bench.

3. Subsequently, the shell membrane was carefully peeled from the yolk membrane. After covering the hole on the egg shell with a culture dish having a diameter of 13 mm, the hatched eggs were re-loaded into a CO ₂ incubator and cultured for 1 day.

4. The compound pellet on a paraffin film was placed on the embryo chorioallantoic membrane of a 4-day-old hatched egg (where there are few existing blood vessels) so that the drug surface adheres to the chorioallantoic membrane. The dose of this compound is 0.12 μmol / egg.

5. The hatched eggs were again loaded into the CO ₂ incubator and cultured for 2 days.

6. About 1 ml of 10% intralipid (angiographic agent) was injected into the chorioallantoic membrane of 6-day-old embryonated eggs, and blood vessel formation around the pellet was observed with a stereoscopic microscope (× 10). The case where no angiogenesis was observed was regarded as positive, and the angiogenesis inhibition rate (%) was calculated by the following formula.

Angiogenesis inhibition rate (%) = [(number of hatched eggs showing positive) / (number of hatched eggs used in the test)] × 100

(Results) The angiogenesis inhibition rate of this compound was 75.
%Met.

From the above, it became clear that the present compound has an angiogenesis inhibitory action.

[0041]

Therefore, it has a chymase inhibitory action.
Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ has an angiogenesis-inhibiting effect, and is associated with angiogenesis, especially diabetic retinopathy, macular degeneration, retinal vein occlusion. , Retinopathy of prematurity,
It is expected to be useful as a prophylactic or therapeutic agent for intraocular angiogenic diseases such as neovascular glaucoma.

─────────────────────────────────────────────────── ─── Continued Front Page (51) Int.Cl. ⁷ Identification Code FI Theme Coat (Reference) A61P 43/00 111 C12N 9/99 // C12N 9/99 A61K 37/64

Claims (5)

[Claims] 1. An angiogenesis inhibitor comprising Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ as an active ingredient. 2. The neovascularization is intraocular neovascularization.
The described inhibitor. 3. The inhibitor according to claim 1, wherein the neovascularization is choroidal neovascularization. 4. An intraocular angiogenic disease therapeutic agent comprising Suc-Val-Pro-HNCH (CH ₂ Ph) P (O) (OPh) ₂ as an active ingredient. 5. The therapeutic agent according to claim 4, wherein the intraocular angiogenic disease is diabetic retinopathy or macular degeneration.