JPH0551565B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to an antiretroviral agent effective in treating various viral diseases caused by retroviruses. [Prior Art and Problems] As research on viruses continues, treatments for viral diseases are gradually being established. HIV (Human
Immuno deficiency virus), HTLV-I (adult T-cell leukemia virus), and the like are known as retroviruses. Retroviruses contain RNA-dependent DNA synthase (hereinafter referred to as reverse transcriptase) within their virus particles.
It is a virus containing After infecting a host cell, RNA is first transcribed into DNA by reverse transcriptase. This DNA is integrated into the host cell chromosome,
then by host cell RNA synthase
mRNA is synthesized. Various viral proteins are produced from this mRNA. There has been a desire to develop a drug that has an epoch-making therapeutic effect on human diseases caused by this retrovirus. [Means for Solving the Problem] The present inventors have conducted research on the retrovirus growth inhibiting effect of various herbal medicines, and have found that baicalein and baicalin, extracted and isolated from the herbal medicine Huang, have retrovirus growth inhibiting effects. I discovered that. The present invention is based on this knowledge and is an antiretroviral agent containing baicalein or baicalin as an active ingredient. Although baicalein and baicalin are known to have anti-inflammatory, anti-allergic, and antihypertensive effects, it has not been previously known that they have anti-viral effects, particularly anti-retroviral effects. Baicalin and baicalein are compounds shown by the chemical structure shown below, and are commercially available from Wako Pure Chemical Industries, Ltd. (However, R=H: baicalein R=GlcA: baicalin) [Effect of the invention] The anti-retroviral effect of the anti-retroviral agent of the present invention will be explained with reference to experimental examples. Experimental Example 1 Cells infected with murine leukemia virus (Raussier strain) were cultured, and the method of Nakajima et al. [CONPARATIVE
LEUKEMIA RESEARCH 1973,
LEUKEMOGENESIS，ED.Y.ITO.AND RM
DUTCHER, UNIV.OF TOKYO PRESS
TOKYO/KARGER, BASEL, PP.603−605
(1975)], reverse transcriptase was isolated and purified. Next, a reaction mixture having the following composition was prepared. Reverse transcriptase 1 unit/ml Polyadenylic acid/oligothymidylic acid complex as template/primer complex [polyadenylic acid (manufactured by Pharmacia): oligothymidylic acid (manufactured by Pharmacia) = 4:1] 2 μg/ml Tris-HCl (PH8. 0) 50mM dithiothreitol 5mM potassium chloride 50mM manganese chloride 0.2mM [ ^3H ]deoxythymidine triphosphate 0.01mM (hereinafter abbreviated as [ ^3H ]dTTP)
(400 cpm/pmol) Glycerol 15 (v/v)% Purified water Appropriate amount * 1 unit means that reverse transcriptase is
It is a unit of specific activity that consumes 1 nmol of dNTP (deoxynucleic acid triphosphate). 20μ of this reaction mixture
Add baicalein and purified water, and baicalin and purified water to make 50μ, [ ³ H]
The incorporation of radioactivity into the acid-insoluble fraction of dTTP was measured using a Beckman scintillation counter, which was defined as reverse transcriptase activity, and the inhibition rate at each concentration was calculated. The results are shown in Table 1.

[Table] Experimental example 2 MT-2 cells were prepared at a concentration of 3 x ¹⁰ cells/ml,
It was cultured in RPMI1640 (manufactured by Gibco) medium containing 10% FCS (manufactured by Gibco). 2 days later, MT-
2 After infecting 1 HIV per 200 cells,
The effects of drug addition were observed by dividing into a group to which baicalein was added at 5 μg/ml and a group to which it was not added (control group). One week later, the number of viable cells was counted, and it was confirmed that the antiretroviral agent of the present invention suppressed MT-2 cell degeneration caused by HIV by 90% or more. This result confirmed an excellent antiretroviral effect. As described above, the antiretroviral agent of the present invention
Since it has the effect of suppressing the proliferation of retroviruses by inhibiting the reverse transcriptase activity necessary for their proliferation, it can be applied to any retrovirus. Specific examples of retroviruses include leukemia virus, sarcoma virus, breast cancer virus, Visna virus, Maedi virus, HIV, HTLV-I, and the like. Next, an acute toxicity test of oral administration of baicalin and baicalein was conducted using ddY male mice and Wistar male rats.
There were no deaths even after oral administration of Kg. Thus, baicalin and baicalein have extremely low toxicity and high safety. Next, the dosage and formulation of baicalin and baicalein will be explained. Baicalin or baicalein can be administered to animals and humans either neat or with conventional pharmaceutical carriers. The dosage form is not particularly limited and may be selected and used as necessary, and may include oral dosage forms such as tablets, capsules, granules, fine granules, and powders, and parenteral dosage forms such as injections and suppositories. It will be done. In order to exert the desired effect as an oral agent,
Although it varies depending on the patient's age, weight, and severity of the disease,
Normally, it is considered appropriate for adults to take 100 to 6000 mg of baicalin or baicalein in divided doses several times a day. In the present invention, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods using, for example, starch, lactose, sucrose, mannitol, carboxymethyl cellulose, cornstarch, inorganic salts, and the like. In addition to the excipients mentioned above, binders, disintegrants, surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents, fragrances, and the like can be used in this type of preparation as appropriate. Specific examples of each are shown below. [Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol. [Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low substituted hydroxypropyl cellulose. [Surfactant] Sodium lauryl sulfate, soybean 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, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate. Baicalin or baicalein can also be administered as a suspension, emulsion, syrup, or elixir, and these various dosage forms may contain flavoring agents and coloring agents. In order to achieve the desired effect as a parenteral agent, it is usually necessary for adults to administer 1 to 100 mg of baicalein or baicalin per day by intravenous injection or intravenous drip, although this will vary depending on the age, weight, and severity of the disease of the patient. note,
Subcutaneous and intramuscular injections are considered appropriate. This parenteral preparation is manufactured according to a conventional method, and diluents generally include distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, mustard oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. can be used. Furthermore, a bactericide, a preservative, and a stabilizer may be added as necessary. In addition, from the standpoint of stability, this parenteral preparation is frozen after being filled into vials, etc.
The liquid formulation can also be reconstituted from the lyophilizate immediately prior to use by removing moisture by conventional lyophilization techniques. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate. Other parenteral preparations include liquid preparations for external use, liniments such as ointments, and suppositories for intrarectal administration, which are manufactured according to conventional methods. Example 1 Cornstarch 44g Crystalline cellulose 40g Carboxymethylcellulose calcium 5g Light anhydrous silicic acid 0.5g Magnesium stearate 0.5g Baicalin 10g Total 100g According to the above recipe, - were mixed uniformly, and compressed and molded using a tablet machine. Tablets of 200 mg were obtained. One tablet of this tablet contains 20 mg of baicalin, and adults should take 5 to 7 tablets a day in several doses. Example 2 Crystalline cellulose 84.5g Magnesium stearate 0.5g Carboxymethyl cellulose calcium 5g Baicalein 10g Total 100g According to the above recipe, a part of and was mixed uniformly, compressed and molded, and then crushed, and the remaining amount of and was The mixture was added, mixed, and compressed using a tablet machine to obtain tablets each weighing 200 mg. One tablet of this tablet contains 20 mg of baicalein, and adults should take 5 to 7 tablets a day in several doses. Example 3 Crystalline cellulose 34.5g 10% hydroxypropyl cellulose ethanol solution 50g Carboxymethyl cellulose calcium 5g Magnesium stearate 0.5g Baicalin 10g Total 100g According to the above recipe, and were mixed uniformly, and then netted by a conventional method, After granulation using an extrusion granulator, drying and crushing, and were mixed and compressed using a tablet machine to obtain tablets each weighing 200 mg. One tablet of this tablet contains 20 mg of baicalin, and adults should take 5 to 7 tablets a day in several doses. Example 4 Corn starch 84g Magnesium stearate 0.5g Calcium carboxymethyl cellulose 5g Light anhydrous silicic acid 0.5g Baicalein 10g Total 100g Mix ~ uniformly according to the above recipe, compression mold with a compression molding machine, and crush with a crusher death,
Granules were obtained by sieving. 1 g of this granule contains 100 mg of baicalein, and adults should take 1 to 2.5 g a day in several doses. Example 5 Crystalline cellulose 40g 10% hydroxypropylcellulose ethanol solution 50g Baicalin 10g Total 100g . After granulation using an extrusion granulator, the mixture was dried and sieved to obtain granules. 1 g of this granule contains 100 mg of baicalin, and adults should take 1 to 2.5 g per day in several doses. Example 6 Cornstarch 89.5g Light anhydrous silicic acid 0.5g Baicalein 10g Total 100g Mix ~ uniformly according to the above recipe,
200 mg was filled into No. 2 capsules. Each capsule contains baicalein
It contains 20 mg, and adults should take 5 to 7 capsules a day in several doses. Example 7 Distilled water for injection Appropriate amount Glucose 200 mg Baicalin 10 mg Total amount 15 ml After dissolving and in distilled water for injection, 5
The mixture was injected into a ml ampoule and autoclaved at 121°C for 15 minutes to obtain an injection.

Claims (1)

[Claims] 1.An antiretroviral agent containing baicalein or baicalin as an active ingredient.