JPH0143740B2 - - 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 succeeded in obtaining a novel isoprenylamine derivative represented by the general formula I 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 general formula [In the formula, R ₁ and R ₂ are

[Formula] (n represents the number 9 or 10) or represents a hydrogen atom, but either one is

[Formula], (alkylene) is a linear or branched lower alkylene which may be substituted with a hydroxy group, and Z represents an optionally substituted heterocyclic group. Isoprenylamine derivatives represented by the general formula (I) and acid addition salts thereof are, for example, (In the formula, n represents the same meaning as above) (e.g. decaprenol, solanesol, etc.) Convert to arylsulfonic acid ester such as sylate,
Then, by reacting an amino compound represented by the general formula H ₂ N-(alkylene)-Z () (in the formula, the alkylene group and Z have the same meanings as above) in the presence or absence of a base. Manufactured. This reaction is usually carried out in an organic solvent. Preferred solvents include common solvents such as methanol, ethanol, chloroform, isopropyl ether, and ethyl acetate. The reaction temperature is suitably in the range of room temperature to 100°C. After the reaction, extraction, concentration, column chromatography,
The desired isoprenylamine derivative can be produced using conventional isolation and purification means such as crystallization. Specific substituents for Z in the amino compound represented by the above general formula () include 2-furyl group, 3-indoyl group, 2-pyrimidylamino group, 10-phenothiazinyl group, 8-(6-methoxy) Examples include isoquinolyl group. Another manufacturing method is to combine the halide or arylsulfonic acid ester with the general formula (wherein alkylene and Z have the same meanings as above, and M represents an alkali metal atom) are reacted and then saponified. This reaction is usually carried out in an aprotic polar solvent such as tetrahydrofuran or N,N-dimethylformamide. The reaction temperature is suitably in the range of room temperature to 100°C. The saponification reaction is suitably carried out in the presence of an alkali (eg, potassium hydroxide, sodium hydroxide, ammonia, etc.) in an alcoholic solvent such as methanol or ethanol, and heated at a temperature ranging from room temperature to 80°C. After the reaction is completed, the desired isoprenylamine derivative can be produced using conventional isolation and purification methods such as extraction, concentration, column chromatography, and crystallization. The acid addition salt of the obtained isoprenylamine derivative can be obtained by mixing the isoprenylamine derivative with a desired acid in, for example, acetone, ethyl acetate, etc., and crystallizing each salt by means such as concentration crystallization. can get. Acid addition salts suitable as pharmaceuticals include salts of hydrochloric acid, acetic acid, citric acid, fumaric acid, lactic acid, and the like. Next, a production example of the isoprenylamine derivative of the present invention will be shown. Production Example 1 N-decaprenylfurfurylamine hydrochloride Add 100 ml of an isopropyl ether solution containing 25.5 g of decaprenyl bromide to 100 ml of an ethanol solution containing 25 g of furfurylamine at room temperature.
After the time-consuming addition, the mixture is stirred at room temperature for 3 hours and heated to reflux with stirring for a further 1 hour. After cooling the reaction solution, add 5% aqueous sodium hydroxide solution to it.
ml and extracted with isopropyl ether. The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. concentrate
26.9 g of the product is treated on a chromatography column packed with 300 g of silica gel using a mixture of chloroform and ethyl acetate as an eluent. N, N from the first eluted section
5.0 g of -didecaprenylfurfurylamine are obtained and 11.2 g of N-decaprenylfurfurylamine are obtained from the next elution section. Dissolve the oily substance of N-decaprenylfurfurylamine in 50 ml of acetone, add hydrogen chloride-ether solution to make it slightly acidic, and leave it in the refrigerator overnight. Separately dry the precipitated crystals and calculate the formula 8.5 g of N-decaprenylfurfurylamine hydrochloride represented by is obtained. Next, the physical properties of this material are as follows. Melting point 82.3-86.3℃ NMR (δ value in CDCl ₃ ) (free base) 7.32 (1H, m) 6.32-6.06 (2H, m) 5.41-4.90 (10H, br) 3.73 (2H, s) 3.20 (2H, d) 1.98 (36H, br-s) 1.58 (33H, s) Elemental analysis value (as C ₅₅ H ₈₆ NO・HCl) Calculated value Actual value C (%) 81.18 80.89 H (%) 10.78 10.92 N (%) 1.72 1.69 Production example 2 2-(3,3-didecaprenylaminopropyl)
Aminopyrimidine 2-(3-aminopropyl)aminopyrimidine
An isopropyl ether solution containing 31.1 decaprenyl bromide in 70 ml of an ethanol solution containing 15.7 g
Add 100ml dropwise at room temperature over an hour while stirring.
The mixture is further stirred at room temperature for 3 hours. Add 100 ml of 5% aqueous sodium hydroxide solution to the reaction mixture, and extract with isopropyl ether. The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. 33.2g concentrate to 350g silica gel
When chromatographed using a mixture of chloroform and ethyl acetate on a chromatographic column packed with 5.7 g of 2-(3,3-didecaprenylaminopropyl)aminopyrimidine represented by is obtained. Next, the physical properties of this material are as follows. n ^28.5 _D = 1.5181 NMR (δ value in CDCl ₃ ) 8.22 (2H, d, J = 5Hz) 6.42 (1H, t, J = 5Hz) 4.9 ~ 5.3 (20H, br) 3.35 ~ 3.65 (2H, m) 3.05 (4H, d, J = 7Hz) 2.50 (2H, t, J = 6Hz) Elemental analysis value (as C ₁₀₇ H ₁₇₂ N ₄ ) Calculated value Actual value C (%) 84.85 84.51 H (%) 11.45 11.22 N (%) ) 3.70 3.54 Production example 3 2-(3-decaprenylaminopropyl)aminopyrimidine dihydrochloride 2-(3-aminopropyl)aminopyrimidine
An isopropyl ether solution containing 31.1 g of decaprenyl bromide in 70 ml of an ethanol solution containing 15.7 g.
Add 100ml dropwise at room temperature over an hour while stirring.
The mixture is further stirred at room temperature for 3 hours. Then 5% to this
Add 100 ml of sodium hydroxide aqueous solution and extract with isopropyl ether. The extract is washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. 33.2g of concentrate silica gel
2-(3,
Elution of 5.7 g of 3-didecaprenylaminopropyl)aminopyrimidine is carried out, followed by elution with a mixture of ethyl acetate and ethanol to obtain 8.5g of 2-(3-decaprenylaminopropyl)aminopyrimidine in the form of an oil. Dissolve this oil in 40 ml of acetone, add hydrogen chloride-ether solution to make it slightly acidic, and leave it in the refrigerator overnight. Separately dry the precipitated crystals and calculate the formula 4.9 g of 2-(3-decaprenylaminopropyl)aminopyrimidine dihydrochloride represented by is obtained. Next, the physical properties of this material are as follows. Melting point 51.6-52.7℃ NMR (in CDCl ₃ , δ value) (free base) 8.20 (2H, d, J = 5Hz) 6.43 (1H, t, J = 5Hz) 4.9-5.3 (10H, br) 3.1-3.7 (4H, m) 2.70 (2H, t, J=6Hz) 2.00 (36H, br) 1.60 (35H, s) Elemental analysis value (as C ₅₇ H ₉₂ N ₄・2HCl・2H ₂ O) Calculated value Actual value C (%) 72.65 72.52 H (%) 10.48 10.46 N (%) 5.94 5.81 Production Example 4 In the same manner as Production Example 3, a halide selected from decaprenyl bromide and solanesyl bromide and 3
-(2-aminoethyl)indole, 10-(3-amino-2-hydroxypropyl)phenothiazine and 8-(4-amino-1-methylbutylamino)-6-methoxyisoquinoline (primaquine)
The following compounds are produced by reacting with a compound selected from: Table 1 shows the physical properties of each compound. In the chemical structural formula in the table below, S represents a solanesyl group, and D represents a decaprenyl group.

【table】

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

【table】

[Table] (2) Effect on influenza virus-infected mice A group of 10 ICR male mice weighing approximately 25 g are infected with influenza virus (PR-8) by nasal spray. The test compound was made into a suspension using a surfactant and administered intraperitoneally 5 times every other day starting 24 hours before virus infection and 2 days after infection (50 mg/day each time).
Kg). 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. Survival number of compound administration group/10 - survival number of surfactant only administration group/10 x 100 = survival rate Table 3 shows the survival rate of each test compound.

【table】

[Table] (3) Toxicity The 50% lethal dose was determined by intravenous administration using 20 to 25 g of ddY male mice. The results are shown in Table 4.

[Table] (4) Human interferon-inducing effect (in vitro) Human-derived normal diploid cells (fibroblast-like cells)
Test compound (ethanol solution and PBS (-)
(diluted with) a suspension with a concentration of 20 n mol,
It was induced according to the method of Edwand A. Havell et al. Using the radioisotope microassay method of H. Ishitsukca et al., the inhibition rate of ³ H-uridine uptake was measured for induced interferon. Table 5 shows the ³ H-uridine uptake inhibition rate of each test compound.

【table】

[Table] (5) Anti-vaccinia virus activity (in vitro) A suspension of the test compound (an ethanol solution and a suspension of 50 nmol in Hanks culture solution) and The viral plaque formation inhibition rate was determined by applying a virus dilution solution. Table 6 shows the inhibition rate of each test compound.

[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 cyst rash, influenza, and 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 depending on the patient's age, symptoms, and course of administration.
It is used several times a day (2 to 4 times) in the range of mg/Kg, preferably 3 to 5 mg/Kg. 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 acacia, gelatin, tragacanth, hydroxypropyl cellulose or polyvinylpyrrolidone, lubricants such as magnesium stearate, talc or silica, disintegrants such as potato starch. It may contain powder, calcium carboxymethylcellulose, or wetting agents such as polyethylene glycol, sorbitan monooleate, polyoxyethylene hydrogenated castor oil, sodium lauryl sulfate, and the like. 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, and fractionated coconut oil such as miglyol. 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, witepsol, 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 Hard capsule for oral use 28 g of 2-(3-decaprenylaminopropyl)aminopyrimidine dihydrochloride and 7.5 g of polyoxyethylene castor oil are dissolved in acetone, and then 25 g of silicic anhydride is mixed. After evaporating the acetone, add 5g of carboxymethylcellulose calcium,
Mix 5g of corn starch, 7.5g of hydroxypropyl cellulose and 20g of microcrystalline cellulose,
Add 30 ml of water, mix and granulate. This was granulated using a No. 24 mesh (BS) granulator (manufactured by Fuji Paudal Co., Ltd.) equipped with a screen. The granules are dried to a moisture content of less than 5% and
No.16 Metsuyu (BS) sieve. Next, these particles are filled with a capsule filling machine.
Filled to 190mg. Formulation example 2 Soft capsule for oral use N-decaprenylfurfurylamine hydrochloride 50g
and polyethylene glycol (macrogol-
400) Mix 130g to make a homogeneous solution. Separately 93g gelatin, 19g glycerin, D-sorbitol
A gelatin solution consisting of 10g of ethyl paraoxybenzoate, 0.4g of ethyl paraoxybenzoate, 0.2g of propyl paraoxybenzoate, and 0.4g of titanium oxide was prepared, and this was used as a capsule coating agent to extract the contents by manual plate punching.
Soft capsules containing 180 mg were manufactured. Formulation Example 3 Injection 5 g of 3-(2-decaprenylaminoethyl)indole, an appropriate amount of peanut oil and 1 g of benzyl alcohol are mixed, and the total amount is made up to 100 c.c. using peanut oil. Dispense 1 c.c. of this solution into ampoules using aseptic technique and seal. Formulation example 4 Injection 2-(3-decaprenylaminopropyl)aminopyrimidine dihydrochloride 1.0g, Nitsukor HCO60
[Nikkol HCO60 (product name)] (hydrogenated castor oil polyoxyethylene-60 mol-ether) 5.0 g,
20g propylene glycol, 10g glycerol,
Mix 5.0g of ethyl alcohol and add distilled water.
Add 100ml and stir. Dispense this solution into 1.4 ml ampoules using aseptic technique and melt and seal.

Claims (1)

[Claims] 1. General formula [In the formula, R ₁ and R ₂ represent [Formula] (n represents the number of 9 or 10) or a hydrogen atom, but either one represents [Formula], and (alkylene) is substituted with a hydroxy group. and Z represents an optionally substituted heterocyclic group]
An isoprenylamine derivative represented by and its acid addition salt.