JPH0688904B2 - Growth inhibitor against virus - Google Patents

Description

TECHNICAL FIELD The present invention relates to an antiviral agent containing neplanocin C as an active ingredient. More specifically, the present invention relates to a growth inhibitor against viruses (excluding human T cell leukemia virus and Epstein-Barr virus) containing neplanocin C or a pharmaceutically acceptable salt thereof as an active ingredient.

Chemotherapy agents including antibiotics that are essentially effective against viruses have not been found so far, but some are known to selectively inhibit the growth of the virus and its metabolic function. . Among these, there is Alra A (Ara-A) Acyclovir as a drug against perpes virus as an antiviral drug, but the only cure for other viral diseases is vaccine therapy. In addition, moroxydine (ABOB, conjunctivitis due to influenza and adenovirus: pool fever), Amantadine (influenza), Iodoxuridine (Idoxuridine, IUDR) (herpes keratitis, herpes varicella), interferon Are known.

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention However, these drugs are also those used for local therapy, those that are still in the development stage, and their definite effects have not been proved. Only the (polio) virus and other few have succeeded, and the fact that many serotypes of the virus can not be expected at present is the current situation, and the development of a drug with a broad antiviral spectrum is awaited. .

The present inventors have completed the present invention as a result of repeated research on agents that overcome the drawbacks of such conventional antiviral agents. Conventionally, neplanocin C has been known as a substance having a growth inhibitory effect on plant pathogenic bacteria and murine leukemia L5178Y cells.
As a result of further studies on the efficacy of this substance, herpes simplex-1 (Herpes simplex-1) virus, herpes simplex-2 (Herpes simplex-2) virus, vaccinia virus (Vaccinia) virus, vesicular stomatitis (Vesicular stomatitis)・ Vesicular stmatitie
s) virus, Reo-1 (Reo-1) virus, Coxsackie B-4 (Coxsakie B-4) virus, Semliki forest virus (Semliki forest) virus, parainfluenza-3
The present invention has been completed by knowing that it exhibits a broad antiviral spectrum against (parainfluenza-3) virus, measles virus, and the like. That is, the present invention relates to a virus growth inhibitor containing neplanocin C as an active ingredient.

As described above, an object of the present invention is to provide an antiviral agent showing a broad antiviral spectrum against various viruses.

Means for Solving the Problems For the purposes of the present invention, neplanocin C is first prepared. For the production method of neplanocin C, see JP-A-55.
No. -24157 discloses a new antibiotic A-11079-B ₂ and a method for producing the same. Nepranocin C in the publication
The physicochemical properties of C ₁₁ H are also described.
₁₃ N ₅ O ₂ , molecular weight 279, melting point 226 ° C (decomposition), soluble in water, soluble in water, dimethyl sulfoxide, dimethylformamide, poorly soluble in notanol, insoluble in ethyl acetate, chloroform, benzene, hexane , A weakly basic substance, and LD _{50 of} neplanocin C
Is 55 mg / Kg (intraperitoneal injection in mice). [For further reference, British Patent No. 2021582B, Current Chemotrapy and Infectious Daisies (Curr
ent Chemotherapy and Infectious Disease), Vol I
I, page 1558 (1980). ] This neplanocin C
Can be produced by the above-mentioned means, for example, Ampierariella S.B.A11079 (Ampullariella s
p.A11079: FERM-P NO.4494).

The neplanocin C in the present invention is a nepranocin C base or a pharmaceutically acceptable salt thereof, for example, an inorganic salt such as hydrochloride or an organic acid salt such as acetate, tartrate, citrate, succinate. Such neplanocin C is prepared as a usual pharmaceutical dosage form, such as an injection, an oral preparation, an ointment, an eye drop, etc., according to the purpose of use. From the antiviral spectrum of neplanocin C, it is most suitable for use as an injectable drug, but eyedrops are also a preferable preparation in view of the large number of viral eye diseases. Such a preparation is usually prepared by aseptically preparing nepranocin C powder, for example, a nepranosine C aqueous solution that has been sterilized with a Millipore filter, and lyophilized powder to be dissolved in distilled water for injection or eye drops before use. To be done. In consideration of compatibility with biological tissue such as osmotic pressure, it is preferable to dissolve the osmotic pressure and ionic strength in physiological saline, Ringer's solution, buffer or the like. Furthermore, as other preparations, suitable preparations such as tablets, powders, granules, capsules, liquids for internal use, suppositories and the like can be prepared according to the administration method by appropriate methods and additives.

A suitable formulation for a dosage form for intravenous injection has a concentration of neplanocin C of 5-10 mg / ml. The dose as an antiviral agent is 50 to 200 mg, preferably 100 mg, per day for an adult, and it may be administered for 1 to 5 days or more.

Effects of the Invention The results of testing the antiviral properties of neplanocin C are described below.

Experimental Example 1 The in vitro antiviral action of neplanocin C was examined.

Herpes simplex-1, -2, using primary rabbit kidney cells
Vaccinia and Vesicular stomatitiv viruses were tested.

100Tc1D _{50 for} monolayer culture of cells using microtrays
(50% infectious titer) of virus was inoculated, and after adsorbing the virus for 1 hour, the virus solution was removed and neplanocin C was included. 3
Culture was performed by adding Eagle's MEM supplemented with% fetal bovine serum. Evaluation of antiviral effect of neplanocin C does not include drug
W containing a drug when the cell degeneration of Well reached 100%
The minimum inhibitory concentration was defined as a 50% dilution of ell cell transformation. The cytotoxicity of neplanocin C on host cells was expressed by comparing the morphological changes with normal cells under a microscope. The experimental results are shown in Table 1.

Experimental Example 2 The in vitro antiviral action of neplanocin C was examined.

Reo-1, Coxsaki using monkey kidney-derived cell line Vero cells
eB-4, Semliki forest, Parainfluenza-3, Measles virus were tested.

100 TclD _{50 for} monolayer culture of cells using microtrays
(50% infectious titer) of virus was inoculated, and after adsorbing the virus for 1 hour, the virus solution was removed and neplanocin C was included. 3
Culture was performed by adding Eagle's MEM supplemented with% fetal bovine serum. Evaluation of antiviral effect of neplanocin C does not include drug
W containing a drug when the cell degeneration of Well reached 100%
The 50% dilution of ell cytopathic was the minimum inhibitory concentration. The cytotoxicity of neplanocin C on host cells was expressed by comparing the morphological changes with normal cells under a microscope. The experimental results are shown in Table 2.

From the above results, neplanocin C shows a broad antiviral spectrum, and between the cytotoxicity and the antiviral activity, 1/20 of herpesvirus, 1/400 of Vaccinia virus, and Parai.
1/30 for nfluenza-3 virus, 1 / for Measles virus
25 clear activity was observed, and adenine arabinoside (A
It can be seen that the spectrum is broader than that of ra-A) and Acycloguanosine (Acyclorir).

As is apparent from the above, nepranosin C is herpes simplex-1, -2 virus, vaccinia virus,
It has antiviral activity against Pesicula stomatisiis virus, Reo-11 virus, Koksatsky B-4 virus, Semliki Forest virus, Parainfluenza-3 virus and Measles virus.

Examples of the present invention will be described below, but the present invention is not limited thereto.

Example 1 1 g of neplanocin C powder was dissolved in 200 ml of sterile distilled water, and aseptically passed through a sterilizing filter to obtain aseptic neplanocin C.
A solution was obtained. Then, 1 ml of this was aseptically dispensed into vials and freeze-dried to prepare a nepranocin C injection preparation. This formulation contains 5 mg of neplanocin C per vial.
Contains. This is distilled water for injection
Dissolve in 1 ml before use.

Example 2 Nepranocin C powder was dissolved in sterile distilled water and then aseptically passed through a sterile filter to prepare a sterile neplanocin C solution, which was aseptically dispensed into an infusion bottle and nepranocin was added.
A lyophilized powder formulation containing 50 mg of C was obtained. This product is dissolved in 100 ml of Ringer's solution for injection to prepare a drip intravenous injection preparation.

Example 3 A lyophilized powder preparation containing 1 mg of neplanocin C is dissolved in 5 ml of sterile physiological saline to prepare an eye drop preparation.

Example 4 A 250 mg tablet having a composition of nepranocin C 50 mg, starch 50 mg, lactose 40 mg, microcrystalline cellulose 105 mg, sucrose fatty acid ester 2.5 mg and magnesium stearate 2.5 mg was prepared.

Claims (3)

[Claims] 1. A herpes simplex-1 virus containing neplanocin C as an active ingredient,
Herpes simplex-2 virus, Vaccinia virus, follicular stomatitis (Vesicular stmatitis, Vesicular stmatiti)
es) virus, Reo-1 virus, Rex-1 virus, Coxsakie B-4 virus, Semliki forest virus, parainfluenza-
3 (Parainfluenza-3) virus or measles (Measle
s) A growth inhibitor for viruses. 2. A growth inhibitor against a virus according to claim 1, wherein the neplanocin C is neplanocin C or a pharmaceutically acceptable salt thereof. 3. Neplanocin C is a microorganism Ampullariella s.
A growth inhibitor against a virus according to claim 1, which is a substance obtained from a culture of p.A11079 (FERM-P No. 4494).