JPH0616549A - Cell proliferation inhibitor of mesangium - Google Patents

Abstract

PURPOSE:To obtain a medicine effective as a therapeutic agent for glomerular diseases represented by glomerular nephritis, nephrotic syndrome, drug renal disorder, etc. CONSTITUTION:A medicine comprises (-)-epicatechin-3-O-gallate of the formula as active ingredient. The compound is obtained from rhizome of Pheum palmatum of crude drug by operation such as extraction, purification, etc. The compound is orally or parenterally administered. In order to exhibit given effects as an oral medicine, 100mg-6g of the compound is administered dividedly several times a day. In order to show given effects as a parenteral medicine, 1-100mg of the compound is applied daily by intravenous drip infusion, hypodermic injection and intramuscular injection.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a drug having an effect of suppressing the proliferation of mesangial cells and useful as a therapeutic drug for glomerular diseases represented by glomerulonephritis, nephrotic syndrome, drug-induced nephropathy and the like. It is a thing.

[0002]

2. Description of the Related Art Currently, drugs effective as therapeutic agents for glomerular diseases represented by glomerulonephritis, nephrotic syndrome, drug-induced nephropathy and the like are corticosteroids,
Immunosuppressants, platelet aggregation inhibitors, etc., but their therapeutic effects cannot be said to be definitive.

In recent years, mesangium has been attracting attention as a cause of glomerular disease. Mesangium is a portion corresponding to the axis of the glomerulus, and is composed of mesangial cells and a mesangial matrix located between the mesangial cells. In particular, in glomerulonephritis, proliferation of mesangial cells is observed in many renal biopsy tissues, and it is considered that the prognosis of glomerulonephritis depends on the degree of proliferation of these cells (kidney and dialysis Vol. 28, Vol. 4 199
0).

Therefore, it has been desired to develop a drug having an effect of suppressing the proliferation of mesangial cells and useful as a therapeutic drug for glomerular diseases.

[0005]

Means for Solving the Problems The inventors of the present invention have conducted extensive studies to solve the above problems, and as a result, It was found that (-)-epicatechin 3-O-gallate represented by the formula (3) has an effect of suppressing the proliferation of mesangial cells.

That is, the present invention is a mesangial cell growth inhibitor containing (-)-epicatechin 3-O-gallate (hereinafter referred to as the drug of the present invention) as an active ingredient.

The drug of the present invention has been conventionally used as an antioxidant. However, it has never been known that this substance has the effect of suppressing the proliferation of mesangial cells.

The drug of the present invention is a crude drug Rhei R
It can be obtained by performing operations such as extraction and purification from Hizoma) or the like, but those commercially available from Funakoshi Chemical Co., Ltd. may be used.

Next, the fact that the drug of the present invention has the effect of suppressing the proliferation of mesangial cells will be described with reference to experimental examples.

Experimental Example Culture of Mesangial Cells The kidney cortex of a normal human obtained by biocrocy was cut into about 1 mm ³ in phosphate buffered saline. Metal strips (No. 60, No. 120, No. 280)
No. No., manufactured by Iida Seisakusho Co., Ltd.) with a silicon spatula. The glomeruli on the 280 mesh were collected.

Next, 20% fetal calf serum (FCS, Arm
our company), 100 μg / ml streptomycin (Meiji Seika Co., Ltd.) containing RPMI1640 (manufactured by Nissui Pharmaceutical Co., Ltd.), and the glomeruli are suspended in a culture dish (diameter 60 mm, manufactured by Corning Co., Ltd.) and spread at 37 °.
C was cultured under the condition of 5% carbon dioxide. The culture medium was exchanged on the 7th day of culturing, and the cells were further cultured for 7 days. On the 14th day of culture, the cells grown around the glomerulus were removed from the culture dish by 0.2
The cells were detached with 5% trypsin (manufactured by Difco) and 0.02% ethylenediaminetetraacetic acid (EDTA, manufactured by Dojindo Co., Ltd.) and subcultured in a culture flask (manufactured by Corning).
After further culturing for 7 days, cells confirmed to be proliferating on the bottom of the culture flask were used as cultured mesangial cells.

Measurement of Cultured Mesangial Cell Proliferation Cultured mesangial cells were made to have a concentration of 10 ⁴ cells / ml (or 10 ⁵ cells / ml), and 100 μl of each was placed on a 96-well flat-bottom culture plate (manufactured by Corning).
Seeded. After culturing for 24 hours and confirming that the mesangial cells are rooted in the plate, the culture solution is aspirated (a
It was sucked up with an uto sucker) and washed once with Hank's solution. After washing, L-Val excluding FCS is DV
2 MEM medium (D-Val MEM) replaced with al
00 μl was added and cultured for 48 hours. After culturing for 48 hours,
Wash once with Hank's solution, add 8% FCS to D-Val
The drug book of the present invention diluted to an appropriate concentration with MEM was added.

Next, the cells were cultured for 24 hours, and ³ H-thymidine (methyl- ³ H-thymidine, Arma was added 6 hours before the end of the culture.
(manufactured by sham) was added at 1 μCi / well. After culturing for 24 hours, the culture solution was replaced with a mixed solution of trypsin and EDTA, and mesangial cells were peeled off. After confirming that the mesangial cells became circular with an actual microscope, the cells were attached on a glass filter using a cell harvester (Labo Mash), and the radioactivity was measured by a Sintillation counter. The results are shown in Table 1.

Table 1

As is clear from the above results, the group to which the drug of the present invention was administered was a non-administration group (the concentration of the drug of the present invention was 0 μg.
/ Ml), the proliferation of mesangial cells was significantly suppressed. Therefore, the drug of the present invention is considered to be useful as a therapeutic drug for glomerular diseases represented by glomerulonephritis, nephrotic syndrome, drug-induced nephropathy and the like.

Next, an acute toxicity test for oral administration of the drug of the present invention was carried out in male ddY mice and Wistar (Wis).
tar) male rats,
There were no deaths even after oral administration of mg / kg.

As described above, it was confirmed that the drug of the present invention has extremely low toxicity and high safety.

Next, the dose and formulation of the drug of the present invention will be described.

The drug of the present invention can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier.
The dosage form is not particularly limited and may be appropriately selected and used as necessary, and may be tablets, capsules, granules, fine granules,
Examples thereof include oral agents such as powders, parenteral agents such as injections and suppositories.

In order to exert a desired effect as an oral preparation, it varies depending on the age, body weight, and degree of disease of the patient, but usually 100 mg to 6 g of the drug of the present invention in an adult
It is considered appropriate to take this in several divided doses a day.

Oral preparations include, for example, starch, lactose, sucrose,
Mannitol, carboxymethyl cellulose, corn starch, inorganic salts and the like are used in a conventional manner.

In this type of preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a flavoring agent, a coloring agent, a flavoring agent, etc. may be appropriately used in addition to the above-mentioned excipients. You can
Specific examples of each are as follows.

[Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch,
Methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, ethyl cellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, carboxymethyl cellulose calcium, carboxymethyl cellulose, low-substituted hydroxypropyl cellulose.

[Surfactant] sodium lauryl sulfate,
Soy lecithin, sucrose fatty acid ester, polysorbate 80.

[Lubricant] Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The agents of the present invention can also be administered as suspensions, emulsions, syrups and elixirs, and these various dosage forms may contain flavoring agents and coloring agents. Good.

In order to exert the desired effect as a parenteral agent, it depends on the age, body weight and degree of disease of the patient.
1 to 100 m per day as a weight of the drug of the present invention in an adult
Intravenous injection up to g, intravenous drip injection, subcutaneous injection, and intramuscular injection seem appropriate.

This parenteral preparation is manufactured by a conventional method, and is generally used as a diluent in distilled water for injection, physiological saline, glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol. Etc. can be used. Further, if necessary, a bactericide, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation may be filled in a vial or the like and then frozen, the water content may be removed by a usual freeze-drying technique, and a liquid preparation may be re-prepared from the freeze-dried product immediately before use. Further, an isotonicity agent, a stabilizer, a preservative, a soothing agent and the like may be added as needed.

Other parenteral agents include external preparations, coating agents such as ointments, suppositories for rectal administration, and the like.
It is manufactured according to a conventional method.

Next, the present invention will be described in more detail with reference to Examples of the pharmaceutical preparation of the present invention, but the present invention is not limited thereto. Example 1 Corn starch 44 g Crystalline cellulose 40 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Magnesium stearate 0.5 g Agent of the present invention 10 g Total 100 g According to the above prescription, are uniformly mixed and compressed with a tableting machine. It was molded to obtain a tablet of 200 mg each. 20 mg of the drug of the present invention is contained in one tablet, and 5 to 7 tablets for adults are to be taken in several divided doses per day.

Example 2 Crystalline cellulose 84.5 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Agent of the present invention 10 g Total 100 g According to the above-mentioned prescription, a part of and was uniformly mixed, compression-molded, and then pulverized. Then, the remaining amounts of and were added and mixed, and the mixture was compression-molded with a tableting machine to obtain 200 mg tablets. 20 mg of the drug of the present invention is contained in one tablet, and 5 to 7 tablets for adults are to be taken in several divided doses per day.

Example 3 Crystalline Cellulose 79.5 g 10% Hydroxypropyl Cellulose Ethanol Solution 50 g Carboxymethyl Cellulose Calcium 5 g Magnesium Stearate 0.5 g Agent of the Invention 10 g Total 145 g The mixture was kneaded by a method, granulated by an extrusion granulator, dried and crushed, and were mixed and compression-molded by a tableting machine to obtain a tablet of 200 mg. 20 mg of the drug of the present invention is contained in one tablet, and 5 to 7 tablets for adults are to be taken in several divided doses per day.

Example 4 Corn starch 84 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Agent according to the present invention 10 g Total 100 g According to the above formulation, 1 to 3 were uniformly mixed and compressed by a compression molding machine. After molding, it was crushed by a crusher and sieved to obtain a granule. 1 g of this granule contains 100 mg of the drug of the present invention.
, 1 to 2.5 g per day for adults is divided into several doses and taken.

Example 5 Crystalline cellulose 86.5 g 10% Hydroxypropyl cellulose ethanol solution 35 g Agent of the present invention 10 g Total 131.5 g According to the above-mentioned prescription, were uniformly mixed and kneaded. After granulating with an extrusion granulator, it was dried and sieved to obtain granules. 1 g of this granule contains 100 parts of the drug of the present invention.
The dosage is 1 to 2.5 g for adults, divided into several doses.

Example 6 Corn starch 89.5 g Light anhydrous silicic acid 0.5 g Agent according to the present invention 10 g Total 100 g According to the above-mentioned formulation, the
Was filled in a No. 2 capsule. One capsule of this capsule contains 20 mg of the drug of the present invention.
Take 5 to 7 capsules several times daily.

Example 7 Distilled water for injection 89.5 g Soybean oil 5 g Soybean phospholipid 2.5 g Glycerin 2 g Drug of the present invention 1 g Total amount 100 g According to the above formulation, was dissolved in and and a solution of and was added to emulsify. Then, an injection was obtained.

Claims (1)

[Claims] 1. The following formula A mesangial cell proliferation inhibitor containing (-)-epicatechin 3-O-gallate represented by