JPS61263919A - Viral multiplication inhibitor - Google Patents

Abstract

PURPOSE:A viral multiplication inhibitor, containing neplanocin C an active constituent and effective against humam T cell leukemic virus considered as the same kind as AIDS causative virus or Epstein-Barr virus which is a causative virus of Burkitt's lymphoma or epipharyngeal cancer. CONSTITUTION:A multiplication inhibitor for human T cell leukemic virus (hereinafter abbreviated to HTLV) or Epstein-Barr virus (hereinafter abbreviated to EBV) containing neplanocin C as an active constituent. The neplanocin C as an antimicrobial agent against plant pathogenic molds and carcinostatic agent is effective as a multiplication inhibitor for HTLV or EBV. The dose thereof is 50-200mg/day, preferably about 100mg/day for adults. Generally, the agent may be prepared as an injection is 5-10ml concentration aseptically prepared in distilled water for injection and intravenously injected. The agent may be prepared as a tablet or suppository.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention is directed to the use of Nep 1anocin (Nep 1anocin).
) T-cell leukemia virus (H) containing C as an active ingredient
umanT-cell Leukemia Viru
s: hereinafter abbreviated as HTLV) or Epstein-Barr Virus
(hereinafter abbreviated as EBV).

Conventional technology: Neplanocin C is used in Ambinilariiella
U-11ariella) belonging to 1% J Ambinillariella nis p. A//07 F (Amp
ulariella sp, A/107ri) (F
ERM-P! The physical properties and chemical structure of a substance named antibiotic A-/107terB2 (later named neplanocin C) found in a culture of a bacterial strain (Japanese Unexamined Patent Application Publication No. 33/1982) have been reported. -211737 publication, British patent number: 20.2/3g2B publication, current
Current Chemotherapy and Infection Diseases
Infectious Disease), Vol.
11, page 155g (79go)]. As for the effects of neplanocin C, it has been reported that it has an antibacterial effect against plant pathogenic filamentous fungi Q, and an anticancer effect showing a life-prolonging effect on tumor cells L-/210r (but not too strong).

On the other hand, human T-cell leukemia is a leukemia associated with immunodeficiency that occurs sporadically in southern Japan and the United States, and HTLV has recently been isolated as the causative virus and confirmed as a human retrovirus. There are three types of HTLV: type ■, type ■, and type ■.Specially, type HTLV1 is the adult T cell leukemia virus (hereinafter referred to as A) that occurs in the common areas of Japan.
It is also found from the abbreviation TLV. In addition, HTL type ■■ is the acquired immunodeficiency syndrome (acquired immunodeficiency syndrome), which is a particular problem in the United States.
AID! The causative virus of S) [Incidentally, lymphadenopathy associated virus
opa also hy-associated virus
) is also known as a virus that has been suggested to be the same. Furthermore, EBV and herpesvirus cluster infections are occurring frequently in Southeast Asia, China, and other countries.

Problems to be Solved by the Invention However, there have been no reports of effective antiviral agents for these HTLV and EBvIl, and currently no treatment has been available. Therefore, the emergence of antiviral agents against HTLV and EBV is awaited.

Means for Solving the Problems As a result of intensive research, the present inventors have surprisingly discovered that neplanocin C is effective against HTLV and EBV and inhibits each of these viruses.

The present invention was completed based on the above findings,
It is a growth inhibitor against HTLV or EBVf containing neplanocin C as an active ingredient.

The object of the present invention is to
The present invention provides an antiviral agent containing neplanocin C as an active ingredient.

First, neplanocin C in the present invention can be produced by the above-mentioned known means, and can be simply produced by Ambinillariella nis p. k/1079 (FERM-P
It can be obtained from a culture of the 'l'lZ&) strain, and is a substance described in the above-mentioned literature, and its structural formula and physical properties are as shown in Tsuko.

[Molecular formula 9 Molecular weight] C+ IHI 3N50□, 279 [Melting point] 224°C (decomposition) [Solubility in solvents] Water, dimethyl) v7. ) V oxide, soluble in dimethylpolamide, slightly soluble in methanol, ethyl acetate, chloroform, pentene, hexane (insoluble in this).

[Distinction between acids and bases]

Weakly basic Neplanocin C is present in its free form or in its salts with inorganic or organic acids, such as hydrochloride, acetate, tartrate,
It may be used in the form of salts such as citrate and succinate.

Preparation of a virus growth inhibitor containing neplanocin C as an active ingredient (in this case, LD5o of neplanocin C)
is 33 to 9 (intraperitoneal injection in mouse), and its effective concentration is more than μf//ml,
As an antiviral agent, the dosage for adults as neplanocin C is about 30-200 m/day, preferably /OOm'
Generally, it can be prepared as an injection solution with a concentration of S~/Q/ml prepared aseptically with distilled water for injection and administered as an intravenous injection. Furthermore, a binder of neplanocin C and an excipient such as lactose, starch, or microcrystalline cellulose, a glucose solution, a gum arabic solution, a seratin solution, water or efnor,
A disintegration aid such as calcium acetate or a surfactant, or a lubricant such as magnesium stearate, calcium stearate, or talc may be appropriately selected and compression molded using the conventional method Q to form a tablet. If necessary, it may be sugar-coated or enteric-coated, and it may also be made into capsules using a conventional method.Furthermore, neplanocin C is added to an aqueous or oily fully open base Q. It may be prepared as a fully-opened product, and in particular, in the case of fully-opened preparation, a fatty acid having a carbon number of g~/g or its sodium salt, N-
It is preferable to add an absorption enhancer such as acyl amino acid, or a hypertonic agent such as chloride or glucose, for example, with reference to British Published Patent Specification No. 2092002 and British Publication No. 209399'1. Also, other powders,
It can be prepared as granules, internal liquids, ointments, creams, aerosols, eye drops, etc., depending on the administration method, using appropriate bases and additives.

The preparation containing neplanocin C as an active ingredient prepared by such Q-filtering is neplanocin CI/)HTLV or E
It may be administered in an effective amount as an antiviral agent against B V +r, for example, for adults, 7 days 30 days or more.

Effect The neplanocin C of the present invention exhibits a good antiproliferative effect against HTLV at a neplanocin C concentration of −μf//ml or less, and also exhibits a good antiproliferative effect on EBV) at a neplanocin C concentration of 2 μf//ml. ．． The effect of inhibiting trowel proliferation was observed;
Particularly, at 20 μf//ml, a good antiproliferative effect is shown, and a preparation containing this neplanocin C as an active ingredient can provide a good antiviral agent against HTLV or EBV.

EXAMPLES Next, the present invention will be specifically described by referring to examples, but the present invention is not limited by these in any way.

Example/(Action against HTLVk) Inhibitory effect of neplanocin C on ATLV (HTLVI type) persistently infected cells A in RPMI-/Otsullo medium supplemented with 10% tallow serum
TLV persistently infected cells M T −/ (Nature (L
ondon), 2911.770~77/, (/9
g/ ), ”Type Virusparticl
es in a cord T-cell 1ine
derived by co-cultivatin
g normal human cord 1
eucocytes and human leul
2 x 105 pieces/
ml of cell suspension was prepared, and neplanocin C solutions of various concentrations prepared in the same medium were added to the cell suspension to prepare a control group without neplanocin C, and 0.0 ml of cell suspension. ．． 2 pfl/ml, 2
p? /ml, 20p? /ml, -? OO
4/'' and pear were cultured at 37°C for S days.

The number of cells was counted using a hemocytometer (improved Neubawell type) once a day starting from the day after culture.

The control group without addition of C grew to 7 x 105 cells in 7 ml on the 2nd day and reached a plateau, but neplanocin 0.20μ?
2×IO' cells/ml were killed on the 7th day. Also, neplanocin C, 200μ? The number of cells/ml also decreased to 10 cells/ml within 7 days.

Sarat Konebranocin 02 μf//ml also takes 5 days in 2 days.
There were 705 x 705 cells in 7 ml, indicating that proliferation was suppressed.

Furthermore, 0.2p of neplanocin C? No effect was observed at a concentration of /ml.

From these results, the concentration of neplanocin C against HTLV is 2 μf/ml or more, especially 20 μf/ml.
A good proliferation-inhibiting effect was observed when the number of cells exceeded /m1.

Example, 21: Effect on EBV] Inhibitory effect of neplanocin C on the proliferation of EBV persistently infected cells RPMI-/OtlIo@JiQco with 10% tallow serum added EBV persistently infected cells P 3 HR-/ (Int , J
.

Cancer, /7, 1123~'12g, (/ri7 Otsu), ``Differential-g-1duction of Epstein-Barr
virus-related
llurmn lymphoblastaid
cell by virus-media also ed
cell-to-cell contactl']2
A cell suspension of x70 cells/ml was prepared, and neplanocin C solutions of various a degrees prepared in the same medium were added to this to give a control group without neplanocin C, and 2 pfl/ml.
, 20 pf//'' each (3
The cells were cultured at 7°C for S days, and the number of cells was measured using a blood pressure counter every 7 days on the day after culture.
Neplanocin C at 20 μfl/ml killed the cells very effectively within 2 days, and neplanocin C
, :l p? /ml also clearly inhibited proliferation compared to the control Q. From this result, neplanocin C
Particularly good growth inhibition effects were observed at 20 μff/ml or more.

Example 3 Neplanocin C/17' was dissolved in a bath of 200 ml of distilled water for injection, sterilized, and filled at a volume of /amp) v/ml, which was lyophilized to /amp/l/, 5+
A neplanocin C-containing injection was prepared.This injection was packaged with 7 ml of a solution for dissolution before use.

Example Gunepura) Shin C! ; Omg, starch so my, lactose 1
10m7, microcrystalline cellulose, i0! rnr9s sucrose fatty acid ester, :z, 3 m'i and 2 30 m with the composition of magnesium stearate λSmg
There were 11 tablets.

Example S A soomy tablet was made with the following composition: Neplanocin Cl 00 mg, starch 10 Ofn'i, lactose go mg, microcrystalline cellulose, 2/On 1 g, sucrose fatty acid ester/l/3 fng and magnesium stearate S~.

[Brief explanation of the drawing]

FIG. 1 shows the growth inhibition curve of neplanocin C against ATLVt, and FIG. 2 shows the growth inhibition curve of neplanocin C against EBV.

Claims (1)

[Claims] (1) A growth inhibitor against human T-cell leukemia virus or Epstein-Barr virus, which contains neplanocin C as an active ingredient.