JPH0499723A - Virus-genomic inactivator - Google Patents

Abstract

PURPOSE:To obtain a virus.genomic inactivator and carcinogenic promoter inhibitor, containing a specific benzylisoquinoline type alkaloid and an analogue compound thereof as an active ingredient and having extremely strong virus genomic inactivating action. CONSTITUTION:A virus.genomic inactivator and carcinogenic promoter inhibitor containing a compound (e.g. cepharanthine or berbamine) expressed by the formula (R1 to R3 are OH or methoxy; R4 and R5 are H or methyl; R6 and R7 are H, OH or methoxy; X is 0; Y is formyl or R1 and R2 together may be -OCH2O-; X and Y together may be -CH2-) as an active ingredient. The compound expressed by the formula is a bisbenzylisoquinoline type alkaloid present in plants of the family Menispermaceae, etc. The dose of the aforementioned compound is preferably 30-300mg/day in the case of oral administration.

Description

[Detailed description of the invention] 3 industrial application fields] TECHNICAL FIELD The present invention relates to a virus genome inactivating agent.

[Conventional technology and issues]

Several viruses, including herpesvirus, exist in the human body in the form of viral genomes, which are activated by some kind of infection and lead to certain types of carcinogenesis and viral diseases. Currently, it is known that substances that promote carcinogenesis are widely present in the environment, and the search for carcinogenic promoter inhibitors to prevent the development of cancer is being carried out widely in plants, but there is still no evidence that they are effective. The number of substances currently available is small, and the development of more effective substances is desired.

[Purpose of the invention]

The present invention searches for a virus/genome inactivating substance that suppresses the expression of the virus/genome by TP△ (tetratylated phorbol ester) in a cultured cell system harboring the EBV (Epstein-Barr virus) genome. The purpose of the present invention is to provide active components as virus/genome inactivators and oncogenic promoter inhibitors.

J Structure of the Invention] As a result of searching for substances that have a virus/genome inactivating effect, the present inventors have found that certain bisbenzyl-noquinoline type alkaloids and their related compounds have extremely strong virus/genome inactivating effects. I found out.

The present invention is based on the general formula (where R1, R2 and R5 are hydroxyl or methoxy groups, R
2 and R5 are a hydrogen atom or a methyl group, R6 and R7 are a hydrogen atom, a hydroxyl group or a methoxy group, X is an oxygen atom, Y is a formyl group, and R1 and R2 together represent 0CH20
-1X and Y should be together (CCH2-te may be present)
This is a virus/genome inactivating agent that contains the compound represented by the following as an active ingredient.

Compounds of formula I include bisbenzylisoquinoline type alkaloids present in plants of the family Cercotaceae and other plants, such as cephalanthine, 2'-norcephalanthine,
Examples include berbamine, tetrandrine, fangquinoline, trilobine, isotrillobine, and secocephalanthine.

The virus/genome inactivating agent of the present invention may use the compound of formula (3) as it is, but usually excipients, binders, lubricants, solvents, stabilizers, etc. are added, and tablets, powders, etc. It is used in formulations such as granules, capsules, injections, and liquids.

Examples of excipients include starch, lactose, methyl cellulose,
Binder such as hydroquinopropyl cellulose, polyvinyl pyro 11 tons, etc., lubricant such as talc, pufferfish stearate/lamb stearate, calcium sulfate stearate, etc. In the case of oral administration, the dosage of the drug according to the invention C) is usually 10 to 1000 mg of the active ingredient per day, preferably 3
0-300 mg.

The inhibitory effect on the expression of the EB virus genome is observed by the following method according to Kozuka et al., published in JP-A-2-101013. The Raji strain culture system, which is a cultured ffB cell derived from Purkitt's lymphoma that contains the EB virus genome, was cultured with the addition of n-acetic acid, which acts synergistically with the oncogenic promoter TAP, and the test substance to promote activity expression. EBV-EA (EB virus early antigen), which is activated by TAP and expressed on the cell surface, is detected by an indirect fluorescent antibody method using antibodies derived from nasopharyngeal cancer patient serum.

Formulation Example 1 Cephalanthine 500 mg, milk 1ffi 3.0 g, corn starch 128 g, hydroxypropyl cellulose 200 mg, and magnesium/ium stearate 20 mg
Mix well, granulate, and tablet to give 100 ml per tablet.
Take the form of mg tablets.

Formulation Example 2 500 mg of HeJl/l/Huff, 25 g of lactose, 1.75 g of potato starch, 240 mg of crystalline cellulose, and 10 mg of carunium stearate were thoroughly mixed, and this mixture was filled into capsules to contain the active ingredients in the capsule. 10m
A capsule containing g.

Formulation Example 3 Tetrandrine hydrochloride 500 mg and D-mannitol 1.0 g are dissolved in distilled water for injection to make a total amount of 100 mf. This solution was filtered through a 0.2μ membrane filter, and 2ml. Dispense into ampoules, melt seal, and heat sterilize to make an injection.

Formulation example 4 Secocephalanthine 500 mg, R sugar 2.5 g,
Potato starch L 75 g, crystalline cellulose 240 g
and 10 mg of calcium stearate were mixed well,
This mixture is filled into capsules containing 10 mg of the active ingredient per capsule.

Formulation example 5 Cephalanthine hydrochloride 500■ and D-mannitol 1
．． Dissolve 0g in distilled water for injection to make a total volume of 100ml'. This solution is filtered through a 0.2μ membrane filter, dispensed into a second ampoule, sealed, and sterilized by heating to obtain an injection.

Test example EB virus latently infected human lymphoblastoid cell line Raji
RPMI 1640 to which fetal serum and antibiotics were added was used as a cell culture medium. Under these culture conditions, the spontaneous expression rate of EBV-EA is 0.1% or less. I×1
Raji cells adjusted to a concentration of 0.06 cells/rd were added to 4 m
M n-butyric acid, 20 ng/ml TPA, and TPA
The cells were cultured at 37° C. for 48 hours in the above culture solution to which cepharanthine was added in a 100-fold molar amount. EBV-EA was stained by indirect fluorescent antibody method using nasopharyngeal cancer patient serum, and the percentage of positive cells was calculated relative to the control to which cephalanthine was not added, and was taken as the inhibitory activity of viral genome expression. The inhibitory activity of cephalanthine on viral genome expression was 63.2%.

The following table shows the results of examining the virus genome expression inhibition activity of the compound of formula (1) using the same procedure but varying the concentration of the test substance.

Claims (1)

[Claims] General formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (I) (In the formula, R_1, R_2 and R_3 are hydroxyl or methoxy groups, R_4 and R_5 are hydrogen atoms or methyl groups, R_6
and R_7 is a hydrogen atom, a hydroxyl group or a methoxy group, X is an oxygen atom, Y is a formyl group, R_1 and R_2 together are -OCH_2O-, X and Y are together -C
A virus/genome inactivating agent containing a compound represented by H_2-) as an active ingredient.