JPH0250111B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel isoprenylamine derivatives and acid addition salts thereof. These compounds are useful in inhibiting viral infections in vertebrates. Substances that have been previously determined to have the effect of preventing or relieving diseases caused by viruses that host vertebrates, or that have been recognized as being able to significantly increase antibody activity and suppress symptoms. There are known substances that have been Reported antiviral substances include interferon, substances that induce interferon (interferon inducer), and synthetic substances that act directly on viral proliferation, such as amantadine hydrochloride or metisazone. be.
Interferon is an antiviral glycoprotein produced by vertebrate cells themselves when they are infected with a virus, and is effective against a wide range of viruses. Inducers for inducing interferon in vertebrates by methods other than viral infection include natural polymeric substances such as the double-stranded ribonucleic acid of certain bacterial phages, or dioxylic acid such as polyinosinic acid-polycytidylic acid. Synthetic polymeric substances such as heavy chain ribonucleic acids, as well as small molecule inducers such as tyrolones, are known. However, interferon has problems in its purification, and a practically economical production method has not yet been developed. Furthermore, conventional interferon inducers have not been put to practical use mainly due to their toxicity. Synthetic antiviral agents currently available on the market that directly act on viral proliferation have a rather narrow range of viral infections that can be treated with them, so the emergence of new synthetic antiviral agents is always desired. For this reason, the present inventors have conducted various studies to find a compound that produces high titer interferon and also has antiviral effects at the animal level. We have succeeded in obtaining a novel isoprenylamine derivative represented by the general formula below and its acid addition salt, which has excellent antiviral and antitumor effects and is therefore expected to be used as a medicine. The novel isoprenylamine derivative according to the present invention has the following general formula: (wherein R ₁ is phenethyl and R ₂ is a group

[Formula], or R ₁ and R ₂ together represent N'-benzylpiperazinyl together with the nitrogen atom to which they are bonded, or 1,7,10,16-tetraoxa- 4,13-diazacyclooctadecanyl, n is a number of 9 or 10). To produce isoprenylamine derivatives represented by the general formula and their acid addition salts, for example, the general formula Isoprenyl alcohol (i.e., decaprenol, or solanesol) represented by (wherein n has the same meaning as above) is converted into a halide (e.g., solanesyl bromide, or decaprunyl bromide) or an aryl sulfonic acid ester (e.g., decaprenyl tosylate or solanesyl tosylate), which is then given the general formula (In the formula, R ₁ and R ₂ have the same meanings as above)
It is produced by reacting the compound represented by in the presence or absence of a base. This reaction is usually carried out in an organic solvent. Such preferred solvents include common solvents such as methanol, ethanol, chloroform, isopropyl ether, benzene, and ethyl acetate. In addition, when performing the above reaction, the amino compound represented by the general formula () is used in large excess, or the amino compound represented by the general formula () is used in the presence of a base (for example, sodium hydroxide, potassium hydroxide, sodium carbonate, or potassium carbonate). Preferably, the reaction is carried out at a temperature of .degree.
After completion of the reaction, the desired isoprenylamine derivative can be produced by treating the reaction solution using conventional isolation and purification means such as extraction, concentration, column chromatography, and crystallization. Furthermore, in the compound represented by the general formula (),
R ₁ is phenethyl and R ₂ is a group

The compound represented by the formula can also be produced by the following method. That is, the above-mentioned halide or arylsulfonic acid ester is reacted with a compound represented by the general formula H ₂ N-R' () (in the formula, R' represents a phenethyl group) in the presence or absence of a base to form a general compound. formula (In the formula, R' and n have the same meanings as above)
A compound represented by is obtained, and the general formula (In the formula, X represents a halogen) by reacting the compound represented by the general formula (In the formula, n and R' have the same meanings as above)
A compound represented by is obtained, and 1 is further added to this compound.
- produced by reacting piperazine-ethanol. In the above reaction, when using 1-piperazineethanol, it is preferable to use this compound in large excess in an alcoholic solvent (for example, methanol or ethanol) or in the absence of a solvent. The reaction temperature is suitably in the range of room temperature to 100°C. After the reaction is complete, the reaction solution is extracted, concentrated,
The desired isoprenylamine derivative can be produced by treatment using conventional isolation and purification means such as column chromatography and crystallization. The resulting acid addition salt of the isoprenylamine derivative can be prepared by mixing the isoprenylamine derivative with the desired acid in a suitable solvent (e.g. acetone or ethyl acetate) and applying means such as concentration, crystallization, etc. Obtainable. Pharmaceutically suitable acid addition salts include, for example, hydrochloric acid, sulfuric acid, phosphoric acid,
Acetic acid, stearic acid, succinic acid, citric acid, tartaric acid, malic acid, oxalic acid, maleic acid, fumaric acid,
Examples include salts such as lactic acid. Next, a production example of the isoprenylamine derivative of the present invention will be shown. Production Example 1 1-Decaprenyl-4-benzylpiperazine To 100 ml of an ethyl acetate solution containing 20 g of N-benzylpiperazine, 100 ml of an ethyl acetate solution containing 25 g of decaprenyl bromide is added dropwise with stirring at room temperature over a period of 2 hours. The mixture was stirred at room temperature for 3 hours and then heated to reflux with stirring for an additional hour. After cooling the reaction solution, 100 ml of 5% sodium hydroxide was added and extracted with ethyl acetate. The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 27.3 g of the concentrate was chromatographed on a chromatographic column packed with 300 g of silica gel using a chloroform-ethyl acetate solution to obtain the formula Oily 1-decaprenyl-4-
10.9 g of benzylpiperazine are obtained. This oil is dissolved in 50 ml of acetone, made slightly acidic by adding hydrogen chloride-ether solution, and left overnight at room temperature. The precipitated crystals were dried separately, and 1-decaprenyl-4-
7.2 g of benzylpiperazine dihydrochloride was obtained. The physical properties of this dihydrochloride are as follows.

【table】

[Table] Production example 2 4-{3-(N-solanesylphenethylamino)
-2-hydroxypropyl}-1-piperazine ethanol trihydrochloride 150 ml of an isopropyl ether solution containing 50 g of solanesyl bromide is added dropwise to 150 ml of an ethanol solution containing 40 g of phenethylamine at room temperature with stirring over a period of 1 hour. The resulting mixture is stirred at room temperature for 3 hours and then heated to reflux with stirring for an additional hour. After cooling the reaction solution, add 150ml of 5% sodium hydroxide aqueous solution.
and extracted with isopropyl ether.
The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. 51.8 g of the concentrate is chromatographed on a chromatography column packed with 550 g of silica gel using a chloroform-ethyl acetate mixture to obtain 31.3 g of oily N-phenethylsolanesylamine. The obtained N-
100 ml of an ethanol solution containing 31.3 g of phenethylsolanesylamine, 30 ml of epichlorohydrin and 30 ml of triethylamine is heated to reflux while stirring for 2 hours. The reaction solution was extracted with isopropyl ether,
Wash with water and saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 31.9 g of concentrate. Next, 50 ml of an ethanol solution containing 9.0 g of this concentrate and 10 g of 1-piperazine ethanol was added to the
Heat to reflux while stirring for an hour. After cooling the reaction solution, add water
Add 200 ml and extract with isopropyl ether. The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. 10.9 g of the concentrate was chromatographed on a chromatography column packed with 150 g of silica gel using a chloroform-methanol mixture to obtain an oily substance 6.1.
get g. Dissolve this oil in 40ml of acetone,
Add ethereal hydrogen chloride solution to make slightly acidic and leave overnight at room temperature. The precipitated crystals were dried separately and the formula 4.3 g of 4-{3-(N-solanesylphenethylamino)-2-hydroxypropyl}-1-piperazine ethanol trihydrochloride is obtained. The physical properties of this material are as follows. Melting point Caramel-like 212℃ (decomposition) NMR (δ value in CDCl ₃ ) (free base) 7.14 (5H, s), 4.82-5.41 (9H, br), 3.43-3.92 (3H, m), 2.98-3.40 (5H, m),
2.40~2.9 (16H, m), 1.98 (32H, br), 1.58 (30H,
s) Elemental analysis value (as C ₆₂ H ₁₀₁ N ₃ O ₂・3HCl・2H ₂ O) C% H% N% Calculated value 69.86 10.21 3.94 Actual value 69.50 10.23 3.82 Production example 3 Bromination in the same manner as Production example 1 decaprenyl and 1,
By reacting 7,10,16tetraoxa-4,13-diazacyclooctadecane, N-decaprenyl-
1,7,10,16-tetraoxer 4,13-diazacyclooctadecane is produced. The physical properties of this material are as follows.

【table】

[Table] Next, the physiological effects of the isoprenylamine derivative of the present invention will be explained in detail. (1) Effect on vaccinia virus-infected mice 0.1 ml of a diluted solution of vaccinia virus was intravenously injected into a group of 10 ICR female mice weighing around 15 g 2 cm from the base of the tail, and 8 days after inoculation, the surface of the tail was injected. The lesions that appeared were stained with a 1% fluorescein-0.5% methylene blue solution and counted. The test compound is made into a suspension using a surfactant and then inoculated with the virus.
A dose of 50 mg/Kg was administered intraperitoneally 24 hours before, and the antiviral effect was evaluated by comparing the number of lesions to a group receiving only surfactant. Table 1 shows the inhibition rate of each test compound.

[Table] (2) Effect on influenza virus infected mice Influenza virus (PR-8) was nasally sprayed into 1 group of 10 ICR male mice weighing approximately 25 g.The test compound was made into a suspension using a surfactant. ,
1 from 24 hours before virus infection and 2 days after infection
The drug was administered intraperitoneally 5 times every other day (50 mg/Kg each time).
Mice that remained alive for 21 days or more after virus infection were considered to be alive, and the survival rate was calculated using the following formula. Number of survivors in the compound administration group/10 - number of survivors in the group administered only with surfactant/10 x 100 = survival rate (%) The survival rates of the test compounds are shown in Table 2.

[Table] (3) Antitumor effect ⁵ tumor cells KN ₇ −85×10 per mouse weigh 20
The drug was administered intraperitoneally to each group of 6 Balb/c male mice around 30 g. The test compound was made into a suspension using a surfactant and administered intraperitoneally three times (30 mg/Kg each time) 24 hours before tumor cell transplantation, and on the 2nd and 5th day after transplantation, and 30 days after transplantation. The antitumor effect was determined by the number of viable cells in the eyes. Table 3 shows the number of survivors for each test compound.

[Table] (4) Toxicity 50% lethal dose was determined by intravenous administration using 20 to 25 g of ddY elastic mice. The results are shown in Table 4.

[Table] (5) Human interferon-inducing effect (in vitro) The test compound (an ethanol solution diluted with PBS (-) at a concentration of 25 nmol) was suspended in human-derived normal diploid cells (fibroblast-like cells). liquid),
Interferon was induced according to the method of Edward A. Havell et al. Using the radioisotope microassay method of H. Ishitsuka et al.
The induced interferon was measured by the inhibition rate of ³ H-uridine uptake. Table 5 shows the inhibition rate of ³ H-uridine uptake of each test compound.

【table】

[Table] As is clear from the above test results, the active ingredient of the present invention not only has the ability to induce interferon in vivo, but also has low toxicity and excellent antiviral action. Furthermore, since the interferon activity and individual antiviral effects of the active ingredient do not necessarily correlate, the antiviral effect of the active ingredient at the animal level is not necessarily limited to interferon, but also to other hosts. It is also possible that an intervening defense mechanism is involved. Many symptoms of diseases caused by viruses are known in humans, such as herpes infections such as herpes simplex and influenza measles.
Therefore, when the active ingredient of the present invention is used for the prevention and treatment of viral infections, it is administered orally, through the respiratory tract, and by subcutaneous, intramuscular, and intravenous injection. The dosage ranges from 0.5 to 20 mg/Kg, preferably from 3 to 5 mg/Kg, and is used several times a day (2 to 4 times) depending on conditions such as the patient's age, symptoms, and route of administration. The active ingredients of the invention can be formulated into compositions for administration in any conventional manner, such as tablets, capsules, granules, powders, etc.
It can be prepared into oral solutions, ophthalmic solutions, suppositories, ointments, injections, etc. When the active ingredient of the present invention is orally administered, it may be formulated into tablets, capsules, granules, or powders. These solid preparations for oral administration contain commonly used excipients, such as silicic anhydride, magnesium aluminate metasilicate, synthetic aluminum silicate, lactose,
Sugar, corn starch, microcrystalline cellulose, hydroxypropyl starch or glycine, binders such as gum arabic, gelatin, tragacanth, hydroxypropylcellulose or polyvinylpyrrolidone, lubricants such as magnesium stearate, talc or silica, disintegrants such as potato starch. , carboxymethylcellulose calcium, or lubricants such as polyethylene glycol, sorbitan monooleate, polyoxyethylene hydrogenated castor oil, sodium lauryl sulfate, etc. In particular, soft capsules can be prepared by dissolving or suspending them in polyethylene glycol or commonly used oil-based bases such as sesame oil, peanut oil, germ oil, fractionated coconut oil such as miglyol, etc. Tablets and granules may be coated in a conventional manner. Oral liquid preparations are aqueous or oily emulsion solutions,
It may be made into a syrup or the like, or it may be a dry product which can be redissolved in a suitable vehicle before use. Such liquid preparations contain commonly used additives such as emulsifying aids such as sorbitol syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, etc., and emulsifying agents such as lecithin, sorbitan monooleate, polyoxyethylene hydrogenated castor oil, non-aqueous Vehicles such as fractionated coconut oil, almond oil, peanut oil, preservatives such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate or sorbic acid may be added. Furthermore, these preparations for oral administration may contain preservatives, stabilizers, etc., if necessary. In addition, when the active ingredient of the present invention is administered in the form of a parenteral suppository, it can be prepared by a conventional method using a lipophilic base such as cacao butter, Huitepzul, or a hydrophilic base such as polyethylene glycol.
Alternatively, a mixture of polyethylene glycol, sesame oil, peanut oil, germ oil, fractionated coconut oil, etc. can be used as a rectal capsule wrapped in a gelatin sheet. The rectal capsule may be coated with a wax-like substance if desired. Next, when this compound is used as an injection, it may be in the form of an oil solution, emulsion, or aqueous solution.
These solvents may contain commonly used emulsifiers, stabilizers, etc. These compositions can contain the compound at one time depending on the method of administration.
% or more, preferably 5% to 50%. Next, examples of formulations of the present invention will be shown. Formulation example 1 Oral hard capsule 1-decaprenyl-4-benzylpiperazine-
25g dihydrochloride and polyoxyethylene castor oil
Dissolve 7.5g in acetone, then 25g of silicic anhydride
Mix. After evaporating the acetone, add 5 g of carboxymethyl cellulose calcium, 5 g of corn starch, and hydroxypropyl cellulose.
Mix 7.5g and 20g of microcrystalline cellulose,
Add ml of water, mix and granulate. this
It was granulated using a No. 24 mesh (BS) granulator equipped with a screen (manufactured by Eckpelletter Fuji Paudal Co., Ltd.). The granules are dried to a moisture content of less than 5% and No.
It was sifted through a 16-metal (BS) sieve. Next, these particles are packed in a capsule filling machine with 190 yen per capsule.
Filled in mg. Formulation example 2 Soft capsule for oral use 4-{3-N-solanesylphenethylamino)-
50 g of 2-hydroxypropyl}-1-piperazine ethanol trihydrochloride and 130 g of polyethylene glycol (Macrogol-400) are mixed to form a homogeneous solution. Separately 93g gelatin, 19g glycerin
g, D-sorbitol 10g, ethyl paraoxybenzoate 0.4g, propyl paraoxybenzoate 0.2g
A gelatin solution containing 0.4 g of titanium oxide and 0.4 g of titanium oxide was prepared, and this gelatin solution was used as a capsule coating agent to produce soft capsules containing 180 mg of content by manual plate punching. Formulation Example 3 Injection N-decaprenyl-1,7,10,16-tetraoxa-4,13-diazacyclooctadecane 1.0 g,
Nikkol HCO60 [Nikkol HCO60 (product name)]
(Hydrogenated castor oil polyoxyethylene-60 mol-ether) 5.0g, propylene glycol 20g,
Mix 10 g of glycerol and 5.0 g of ethyl alcohol, add 100 ml of distilled water, and stir. Dispense this solution into 1.4 ml ampoules using aseptic technique and melt and seal.

Claims (1)

[Claims] First-order general formula (where R ₁ is phenethyl and R ₂ is the group [formula], or R ₁ and R ₂ together together with the nitrogen atom to which they are bonded are N'-benzylpipe or 1,7,10,16-tetraoxa-4,13-diazacyclooctadecanyl, n is a number of 9 or 10); its acid addition salts.