JPH1179992A - Apoptosis suppressant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an apoptosis suppressant effective for liver diseases, neurodegenerative diseases and inflammatory diseases by formulating a specific crude medicine extract as an active ingredient. SOLUTION: This apoptosis suppressant comprises lycorine represented by the formula or its pharmacologically permissible salt as an active ingredient. Lycorine represented by the formula is a component for Sekisan which is an emetic agent and is one of Amaryllidaceae alkaloids contained in amaryllidaceous plant. The dose of lycorine of the formula or its pharmacologically permissible salt is normally 0.1-20 mg, preferably 1-10 mg/kg weight as lycorine hydrochloride.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an apoptosis inhibitor containing lycorine or a pharmacologically acceptable salt thereof as an active ingredient, and a disease effective in inhibiting apoptosis, for example, liver disease such as cirrhosis, Alzheimer's disease and the like. A therapeutic or prophylactic agent for neurodegenerative diseases and inflammatory diseases.

[0002]

2. Description of the Related Art Apoptosis is the death of cells caused by the activation of a cell self-destruction program controlled by the genes of the cells. Apoptosis is found, for example, in organ development, metamorphosis, ridge formation, cell replacement of epidermal cells, and the like. The form of cell death, apoptosis, is achieved by cytoplasmic enrichment, loss of plasma membrane hair, nuclear segmentation, and cleavage of chromosomal DNA at nucleosome units (The Journal of Biochemistry, 267, 10709,
1992). The reasons why apoptosis is attracting attention in relation to diseases are (1) involvement in the elimination of autoreactive immunocompetent cells, (2) involvement in the reduction of T cells by AIDS virus, (3) Alzheimer's disease, etc. And (4) spontaneous disappearance of tumor cells in cancer lesions and cell death by anticancer agents.

[0003] As substances that inhibit apoptosis, inhibitors of proteases that mediate apoptosis (Nature,
370, 270, 1994), berberine, palpatin,
Chinese herbal components such as capillarisin (Japanese Patent Application Laid-Open No. 9-30983) and substances in cell cultures (Japanese Patent Application Laid-Open No. 9-67260) are known.
However, these compounds have problems such as an excessively high effective concentration and not being specific for apoptosis, and there is a need for a more effective and highly specific inhibitor of apoptosis.

[0004] From the above, no drug having a practically sufficient effect as an apoptosis inhibitor has been developed.
Therefore, at present, no therapeutic or preventive agent for a disease effective for inhibiting apoptosis having a practically sufficient effect has been developed.

[0005]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above-mentioned prior art, and is intended to suppress apoptosis which is different from drugs for treating cirrhosis, drugs for treating Alzheimer's disease, and anti-inflammatory drugs which have already been put to practical use. An object of the present invention is to provide an apoptosis inhibitor having a new mechanism of action, and a therapeutic or preventive agent for a disease for which apoptosis inhibition is effective, for example, a therapeutic agent for cirrhosis, a therapeutic agent for Alzheimer's disease, an anti-inflammatory agent, and the like.

[0006]

Means for Solving the Problems The present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, it has been found that lycorine or a pharmacologically acceptable salt thereof suppresses apoptosis.
The present inventors have found that cell death that is inconvenient to the living body, for example, cell death during cirrhosis or degeneration of nerve cells, is prevented, and the present invention has been completed.

That is, a first aspect of the present invention is that the formula (I):

[0008]

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An apoptosis inhibitor containing, as an active ingredient, lycorine or a pharmacologically acceptable salt thereof represented by the following formula (I):

[0010]

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[0011] The present invention relates to a therapeutic or prophylactic agent for diseases in which suppression of apoptosis is effective, which comprises lycorine represented by the formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
Lycorine used in the present invention is a component of the emetic agent Shigaru (Sexan) (Japanese Clinical Supplement.
Amarylli in Amaryllidaceae plants (Page 1991)
daceae alkaloids. Has protein synthesis inhibitory activity (Jounalof Biological Chemistry. 261, 505,
1986), antiviral activity (Jounalof Natural Produc
ts. 55, 1569, 1992), and an immunosuppressive effect (German Patent DE 3426109 A1 or JP-A-61-36221). A document describing the pituitary-mediated anti-inflammatory effect of the herbal medicine sexan containing lycorine (Yao Hsueh Hsueh
Pao, 12, 767, 1965), but the apoptosis inhibitory effect of lycorine was not known.

The present inventors have searched for various crude drug extracts in order to find compounds having an apoptosis-suppressing action. As a result, they have found that an active substance is present in lacunthai, purified and isolated the active ingredient, and found that lycorine was active. The present invention was completed by identifying the components. The pharmacologically acceptable salts used in the present invention include acetate, sulfate, hydrochloride and the like.

Further, the present invention provides a compound of the formula (I):

[0015]

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A therapeutic or prophylactic agent for diseases effective in suppressing apoptosis, comprising as an active ingredient lycorine represented by the formula (I) or a pharmaceutically acceptable salt thereof. Examples of the disease in which the suppression of apoptosis is effective include, for example, cirrhosis, liver disease such as acute or chronic hepatitis, Alzheimer's disease,
Alzheimer's dementia, neurodegenerative diseases such as Parkinson's disease, inflammatory diseases such as rheumatoid arthritis, ulcerative colitis, and nephritis, but by suppressing apoptosis, it is possible to prevent undesired cell death for the living body. The disease is not limited to these as long as it can be caused.

The apoptosis inhibitor of the present invention and the therapeutic or prophylactic agent for diseases for which apoptosis inhibition is effective are added with a carrier, a stabilizer, an absorption enhancer and the like, if necessary, to give tablets, powders and granules. , Capsules, syrups and the like, or suppositories, injections, external preparations,
It may be administered parenterally as drops or the like. The dose of the lycorine represented by the formula (I) of the present invention or a pharmaceutically acceptable salt thereof in the apoptosis-suppressing agent of the present invention and the therapeutic or prophylactic agent for a disease in which apoptosis suppression is effective is as follows:
It can be appropriately adjusted depending on the disease to be treated and prevented, the patient's age, body weight, and the like. Usually, it is 0.1 mg to 20 mg, preferably 1 mg to 10 mg, per 1 kg of body weight as lycorine hydrochloride. Preferably, the number of administrations is determined.

[0018]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the present invention. Example 1 Inhibitory Effect of Lycorine on Apoptosis of MM46 Cells by Calprotectin MM46 cells derived from mouse breast cancer (purchased from American Type Culture Collection) were spread on a 96-well plate at 1 × 10 ⁴ cells / well, and then And calprotectin derived from rat polymorphonuclear leukocytes (adjusted from polymorphonuclear leukocytes in a conventional manner) and RP containing 5% fetal calf serum
When cultured at 37 ° C. in MI-1640 medium (Dainippon Pharmaceutical Co., Ltd.), MM46 cells apoptotic by calprotectin appeared after 16 hours. In order to examine the apoptosis-suppressing effect of lycorine, lycorine hydrochloride (manufactured by LATOXAN) was added simultaneously with addition of calprotectin at 0, 0.2, and 1.0 μg / ml, and after 20 hours, the number of surviving cells was measured by MTT assay. It was measured. As a control, the same measurement was carried out when calprotectin was not added. Table 1 shows the percentage of living cells obtained by subtracting the absorbance at 630 nm from the absorbance at 588 nm in each well.

[0019]

[Table 1]

As shown in Table 1, lycorine hydrochloride 1 μg / m
It was found that the addition of l suppressed the apoptosis of MM46 cells, and the MTT reduction ability of the cells was closer to the group without addition. That is, ricolin inhibited apoptosis caused by calprotectin derived from polynuclear leukocytes. Example 2 Effect of Lycorine on TNF Production of Macrophages Macrophages derived from mouse peritoneal cavity (prepared from mice in a conventional manner) were spread on a 96-well plate at 1.2 × 10 ⁵ cells / well, and then lycorine hydrochloride (LATOXA).
N) were added at 0, 0.04, 0.2, and 1.0 µg / ml, respectively, and cultured at 37 ° C for 3 hours. After that, LPS (Sigma:
(1 μg / ml) was added as a stimulant, and after 2 hours, TNF activity released into the culture supernatant was measured by a cytotoxicity test using L cells (purchased from American Type Culture Collection). As a control, TNF activity without LPS was measured in the same manner. Table 2 shows the results.

[0021]

[Table 2]

As shown in Table 2, LPS-stimulated TNF production was suppressed by lycorine.

[0023]

Industrial Applicability According to the present invention, by suppressing apoptosis, it is possible to prevent undesired cell death such as cell death during cirrhosis and degeneration of nerve cells. The present invention can provide an apoptosis inhibitor having the same, and a therapeutic or prophylactic agent for a disease for which apoptosis inhibition is effective, for example, a therapeutic agent for cirrhosis, a therapeutic agent for Alzheimer's disease, an anti-inflammatory agent and the like.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 31/47 ACL A61K 31/47 ACL ACS ACS ACS ADS ADS AGZ AGZ // A61K 35/78 35/78 C C07D 491/16 C07D 491 / 16

Claims (3)

[Claims] 1. A compound of formula (I): An apoptosis inhibitor comprising, as an active ingredient, lycorine represented by the formula (I) or a pharmaceutically acceptable salt thereof. 2. Formula (I): A therapeutic or prophylactic agent for a disease in which suppression of apoptosis is effective, comprising lycorine represented by the formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. 3. The therapeutic or preventive agent according to claim 2, wherein the disease is a liver disease, a neurodegenerative disease, or an inflammatory disease.