JPS63287726A - Anti-aids virus agent - Google Patents

Abstract

PURPOSE:To obtain the titled preparation comprising a compound containing a hypoxanthine base or a salt thereof as an active ingredient. CONSTITUTION:An anti-AIDS virus agent comprising 0.01-100wt.%, preferably 1-70wt.% compound shown by the formula or salt thereof, a carrier (e.g. liquid carrier such as water, alcohol or soybean oil or solid carrier such as saccharide, for instance maltose or amino acids) and other auxiliaries. In the case of injection, the injection contains 0.01-20wt.%, preferably 0.1-10wt.% compound shown by formula. Tablet, capsule, powder, granule, solution, dry syrup, etc., may be cited as the form of oral drug. A dose is 1-800mg/kg, has low toxicity and small accumulation of toxicity by continuous administration.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to an anti-AIDS virus agent.

[Conventional technology]

Acquired immunodeficiency syndrome (hereinafter referred to as AIDS) was discovered to be a disease caused by a virus (hereinafter referred to as AIDS virus)1.For example, LAV was isolated at the Institut Pasteur in France, HTLV-■ has been isolated in the United States of Health (these viruses are currently classified as
An engineering mmu -nodeficiency Viru
It has been internationally proposed to call it s). However, no effective preventive treatment for AIDS caused by infection with H-V has yet been found.

[Problems to be solved by the present invention]

Therefore, it is desired to develop a new anti-AIDS virus agent that is highly effective in suppressing the AIDS virus and has low toxicity.

[Failure to solve the problem]

As a result of various studies, the inventors found that a new compound having a hypoxanthin base represented by the structural formula or a salt thereof (hereinafter referred to as the present compound) has an anti-AIDS virus effect and can be used as an anti-AIDS virus agent. I found it.

The present invention has been completed based on the above findings. For example, this compound acts on a compound having an adenine base represented by the following formula (2) (hereinafter referred to as oxetanosine) with adenosine deaminase or a microbial culture having a similar ability, a processed product thereof, or a sample from an animal tissue. By doing so, the following formula (
1) The present compound having a hyboxanthin base represented by the following can be produced. Oxetanosine, which has the structural formula '(2) of the raw material compound, is a well-known compound that was presented at the Japanese Society of Agricultural Chemistry, 1986 conference abstracts (Kyoto) on page 291, title A3 -58, and was published in Japanese Patent Application Laid-Open No. 1986-58. -29399
2 is also disclosed.

The present compound forms a salt with an acid, but the acid used to form the salt may be any pharmacologically acceptable acid.
For example, hydrochloric acid, sulfuric acid, phosphoric acid, etc. are preferred.

The anti-AIDS virus agent of the present invention is administered as an injection, an oral preparation, a skewer, or the like using the present compound alone or by mixing the present compound with an excipient or carrier.

Pharmaceutically acceptable excipients and carriers are selected, and their types and compositions are determined by the route and method of administration. For example, water as a liquid carrier.

Alcohol or soybean oil, peanut oil, sesame oil.

Animal and vegetable oils such as mineral oil, and still synthetic oils are used.

Sugars such as maltose and sucrose as solid carriers,
Amino acids, hydroxypropylcellulose natocellulose derivatives, organic acid salts such as magnesium stearate, etc. are used. For injections, generally physiological saline, various buffer solutions, glucose, inositol, mannitol'
Glycols such as 4o sugar M solution, z-tyrene glycol, and polyethylene glycol are preferable. Also, inositol, ma/nitol, crucose, and mannose.

It is made into a lyophilized preparation with excipients such as sugars such as maltose and sucrose, and amino acids such as phenylalanine, and is then administered in a suitable solvent for injection such as sterile water, physiological saline, glucose solution, electrolyte solution, and amino acids. It can also be administered by dissolving it in a liquid for intravenous administration such as.

The content of this compound in the preparation varies depending on the preparation, but is usually 0.01 to 100% by weight, usually 0.1 to 90%,
It is preferably 1 to 70% by weight, with the remainder consisting of carriers and other auxiliary agents commonly used in pharmaceuticals. For example, in the case of an injection solution, 1 is usually 0.01 to 20% by weight, preferably 0.
．． It is preferable to contain 1 to 10% by weight of real metal. When administered orally, tablets, capsules, powders, granules, liquids,
It is used in the form of dry syrup, etc. In the case of capsules, tablets, granules, and powders, the content of the compound is generally about 3 to 3.
It is 100% by weight, preferably 5 to 90% by weight, with the remainder being carrier.

The dosage depends on the patient's age, weight, symptoms, and therapeutic purpose.

Although determined by the administration method, the therapeutic amount is generally 1 to 800 m9/KP for parenteral or oral administration.

The acute toxicity value (LD5o) of this compound is 2001 in mice.
119/yield (1v) or more, and has low toxicity.

It is also characterized by low toxicity accumulation upon continuous administration.

Next, examples of formulations of the present invention will be shown.

Formulation Example 1 Purified water was added to 30 parts by weight of the hydrochloride of compound (1) to make a total of 2000 parts, and the solution was dissolved and filtered for sterilization using a Millipore filter GS type.

This tear fluid 2? The mixture was placed in a 10 ae vial and lyophilized to obtain a lyophilized injection containing 30 mg of the hydrochloride of compound (1) per vial.

Formulation Example 2 Granule Compound (1) 50 parts by weight, 600 parts of lactose, 360 parts of crystalline cellulose and 20 parts of hydroxypropyl cellulose were thoroughly mixed, and the mixture was thoroughly mixed using a roll compactor.
①), crush the mixture, and sieve it to a size between 16 mesh and 60 mesh to obtain granules.

Formulation Example 3 Tablets 30 parts by weight of Compound (1), 120 parts of crystalline lactose, 147 parts of crystalline cellulose and 3 parts of magnesium stearate were compressed using a ■-type mixer to obtain tablets each weighing 300 mg.

Next, the anti-AIDS virus effect of this compound will be specifically explained using test examples.

Test example Human engineering mmunodeficiency V
Approximately 50,000 MT-4 cells/ml were placed in a 24-well tray, 100 μt of a solution containing a certain amount of the compound of the present invention was added, and the mixture was incubated at 37°C at -5% (v/v).
After culturing in a carbon dioxide gas incubator for 2 hours,
~104 infectious units were added, and after culturing for 4 days, a portion of the culture solution was smeared onto a slide glass, fixed with acetone, and then the expression of viral antigen was observed using a fluorescent antibody method.

The primary antibody for the fluorescent antibody method was serum from an AIDS patient, and the secondary antibody was anti-human 17G labeled with FI-TC.
was used.

1. Cell degeneration of M']''-4 cells by the compound of the present invention was carried out without adding virus and observed under a microscope.

Activity of the compound of the present invention against H-V: 100±~+2 The compound of the present invention was used after being dissolved in DMSO. Viral antigen expression with DMSO alone was 80-90%.

As is clear from the above, the compound of the present invention has a remarkable growth-inhibiting effect on H. lactis and causes little cell degeneration, so it is expected to be an anti-AIDS therapeutic agent.

Reference Example 1 (Production of the Present Compound) Compound 78 of formula (2) was added to M/10 phosphate buffer 59.
1, and 25 μt (9.9 units) of adenosine deaminase (Sigma Co., Ltd. %FIC3,5,4,4) was added thereto, and the mixture was reacted at 25° C. for 5 hours with stirring. Next, heat at 100°C for 5 minutes.

After stopping the reaction, apply one reaction solution to MC Gθ1■CHP
The product was passed through a column packed with 50mJ of 20F to adsorb it, and after washing with water, 150ml of 20% water-containing methanol was added.
By eluting with V, concentrating and drying, a colorless, powdery compound 69 (yield: 88.1%) of formula (1) was obtained.

The compound of formula (1) obtained here, q (C2'R
The physical and chemical properties of 13'R,4's)-3',4'-bis(hydroxymethyl)-2'-oxetanyl]-hypoxanthine are shown below.

(1) Appearance Colorless powder (2) Elemental analysis value (%) (C10H1204N4
)CHON Theoretical value 47.62 4.80 25.37 22.2
1 Actual value 47.41 4.96 25.58 22
．． 05(3) Molecular formula (molecular weight) C10H1204N4 (252,24)(4)
Melting point: 203-205°C (5) Specific rotation (a:) 20=-a, 8° (C=0.4 s
5+ H2O) (6) Ultraviolet absorption spectrum The ultraviolet absorption and molecular extinction coefficient values in water, 0.1N hydrochloric acid and 0.1N sodium hydroxide solution are respectively iaX (1og9) = 2 4 8.5 n
m (4,10)0, I NHcl λmaX (logε) = 249 nm (4,
08) 0.1N NaOH 8maX (Logε) = 253 nm (4,
+2) (7) Infrared absorption spectrum The absorption maximum value (wave number --1) in the infrared absorption spectrum measured on a potassium bromide tablet is shown below.

16951590155215151492(sh)+
465([]h)+4501420137613401
320 (day h) 12so (sh) 1220115011
20 (Sunday h) +1001050 (Elh) 101297
[190084578B 720 (day h)
685(8) Solubility in solvents Soluble in water, methanol, dimethyl sulfoxide,
Propertool Insoluble or sparingly soluble in organic solvents such as acetone, ethyl acetate, ether and benzene.

(9) The color reaction Lydon-Smith reaction and 10-thiosulfuric acid reaction are positive, and the ninhydrin reaction is negative.

(10) Rf value of thin layer crostography Fusilica gel thin layer Kieeel gem 60 F 254°o,
n-butanol:acetic acid:water (4:1:2) and n-butanol 82
Rf: 0.43 and 0.10 are shown by developing with each developing solvent system of 8% aqueous ammonia:water (1o:o, s:1).

(11) H-nuclear magnetic resonance spectrum was measured using tetramethylsilane in deuterated dimethyl sulfoxide as an internal standard.
As shown in the figure.

(12) 13C-Nuclear magnetic resonance spectrum 13C internalized using dioxane in heavy water (δ67.4) as an internal standard
-The chemical shift (δ-value) of the nuclear magnetic resonance spectrum is 159.7 149.2 147.3 141・
3 124.582.8 BQ, 4
63.3 59.8 45.5 Patent applicant Nippon Kayaku Co., Ltd. = 13 U -

Claims (1)

[Claims] The following formula (1) ▲ Numerical formulas, chemical formulas, tables, etc. ▼ AIDS virus agent.