JPH0128737B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel isoprenylamine derivatives, acid addition salts thereof, and antiviral agents containing the same. These compounds are useful as pharmaceuticals or intermediates thereof for 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 are interferons, substances that induce interferons (interferon inducers), and synthetic substances that act directly on viral proliferation, such as amantadine hydrochloride or metisazone. . 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 general formula [In the formula, n is 2 to 10, A and B are each a hydrogen atom, or A and B together form a single bond, and n is 4
in which case A and B may be a combination of the above two cases, R is hydrogen, benzyl or lower alkyl, p is 2 or 3 and q
is at least 2]. To produce isoprenylamine derivatives represented by the general formula and their acid addition salts, for example, (In the formula, A, B and n have the same meanings as above)
An isoprenyl alcohol represented by (e.g. decaprenol, solanesol, phytol or geraniol) is prepared by a known method from a halide (e.g. geranyl bromide, solanesyl bromide, phytyl bromide or decaprenyl bromide) or an aryl sulfonic acid ester (e.g. decaprenyl tosylate or solanesyl tosylate) and then the general formula () (In the formula, R, p and q have the same meanings as above.) It is produced by reacting a compound represented by the following in the presence or absence of a base. This reaction is usually carried out in an organic solvent. Preferred solvents are common solvents such as methanol, ethanol, chloroform, isopropyl ether, benzene, and ethyl acetate. The amino compound represented by the general formula () is used in large excess or is reacted in the presence of a base (for example, sodium or potassium hydroxide or sodium or potassium carbonate) at a temperature from room temperature to 100°C. After completion of the reaction, the desired isoprenylamine derivative can be prepared using conventional isolation and purification means such as extraction, concentration, column chromatography or crystallization. Alternatively, as another production method, when R is benzyl or lower alkyl in the compound represented by the general formula (), the general formula () can be prepared using the same reaction conditions as above. (wherein A, B, n, p and q have the same meanings as above), and then a compound represented by the general formula R'COX () (wherein R' is methyl or phenyl and X is a halogen atom) in the presence of a base (e.g. pyridine or a tertiary amine such as triethylamine) at a temperature of 0°C to 50°C.
By reacting with the general formula () (In the formula, R' represents methyl or phenyl, and A, B, n, p, and q have the same meanings as above.) A reducing agent (for example, lithium aluminum hydride) is added to this compound. and react in an organic solvent. Preferred solvents used in the reaction include ether and tetrahydrofuran. Reaction temperature ranges from room temperature to
A range up to 60°C is preferred. After the reaction, extraction,
The desired isoprenylamine derivative can be produced from the reaction solution using conventional isolation and purification means such as concentration, column chromatography, and crystallization. The resulting acid addition salt of the isoprenylamine derivative can be obtained by mixing the isoprenylamine derivative with the desired acid in a suitable solvent (eg, acetone or ethyl acetate) and concentrating or crystallizing the mixture. As a medicine, suitable acid addition salts include, for example, 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-Decaprenyltriethylenetetramine To 100ml of a chloroform solution containing 47.0g of triethylenetetramine, 100ml of a chloroform solution containing 40g of decaprenyl bromide was added dropwise with stirring at room temperature for 1 hour. Stir for an hour. The reaction solution was concentrated under reduced pressure, and the concentrate was extracted with ethyl acetate. The extract was washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 41.4 g of a concentrate. The concentrate is then dissolved in 100 ml of isopropyl ether and 20 g of soda carbonate are added to the resulting solution. While cooling with ice water and stirring, 30 ml of trifluoroacetic anhydride was added dropwise over a period of 1 hour, and the mixture was further stirred for 3 hours while cooling. After separating the insoluble matter from the reaction solution, it is concentrated under reduced pressure. Add about 50 ml of benzene and further concentrate under reduced pressure. Using a chromatography column packed with 450 g of silica gel, 43.9 g of the concentrate was eluted with a mixture of benzene and ethyl acetate to obtain N-decaprenyl-N,
10.1 g of N′,N″,N-tetratrifluoroacetyltriethylenetetraamine is obtained.
Decaprenyl-N,N′,N″,N-tetrafluoroacetyltriethylenetetraamine 10.1g
Add 100 ml of a 10% potassium hydroxide solution in ethanol, and heat under reflux for 1 hour. Add 300 ml of water to the reaction solution and extract the mixture with ethyl acetate. The extract was washed with water and saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the formula 9.5 g of N-decaprenyltriethylenetetramine represented by is obtained as an oil. The physical properties of this material are as follows. n ^28.0 _D = 1.5109 NMR (in CDCl ₃ , δ value) 4.9-5.3 (10H, br) 3.20 (2H, b J=7Hz) 2.72 (12H, s) 2.00 (36H, br) 1.60 (33H, s) Element Analytical value (as _C56H98N4・_2H2O ) Calculated value Actual value C (%) 77.90 77.98 H (%) 11.91 11.75 N ₍ %) 6.49 6.36 Production example 2 N-decaprenyl- _N , N', N″,N-tetrabenzyltriethylenetetramine tetrahydrochloride Chloroform solution containing 5.0g of N-decaprenyltriethylenetetramine obtained in Production Example 1 50
10 ml of pyridine is added to 10 ml of pyridine, and 30 ml of a chloroform solution containing 4.5 g of benzoyl chloride is added dropwise thereto over 1 hour with stirring while cooling in an ice bath, and the resulting mixture is further stirred at room temperature for 2 hours. The reaction solution is extracted with isopropyl ether, and the extract is washed successively with water, 5% hydrochloric acid, 5% aqueous sodium bicarbonate solution, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 6.7 g of this concentrate was chromatographed on a chromatography column packed with 100 g of silica gel using a mixture of chloroform and ethyl acetate as the eluent to obtain N-decaprenyl-N,N',N'',N-tetrabenzoyltriethylene. Obtain 4.2 g of tetramine.Next, add 2.0 g of lithium aluminum hydride in a small amount to 50 ml of an anhydrous diethyl ether solution of 4.2 g of N-decaprenyl-N,N',N'',N-tetrabenzoyltriethylenetetramine while stirring at room temperature. Add one by one. After the addition was completed, the mixture was stirred at room temperature for 1 hour, and then heated under reflux for 3 hours while stirring. 10 to the reaction solution
Add 100 ml of % aqueous sodium hydroxide 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. Dissolve 3.8 g of the concentrate in 100 ml of acetone, add hydrogen chloride-ether solution to make it slightly acidic, and then concentrate the solvent to dryness under reduced pressure to obtain the formula N-decaprenyl-N, N′, N″,
N-tetrabenzyltriethylenetetramine 4
3.8 g of hydrochloride are obtained. The physical properties of this material are as follows. Melting point Caramel NMR (δ value in CDCl ₃ ) (Free) 7.13 (20H, s) 4.9-5.3 (10H, br) 3.46 (2H, s) 3.40 (6H, br-s) 2.90 (2H, d J=7Hz ) 2.2~2.6 (12H, m) 2.00 (36H, br) 1.60 (33H, s) Elemental analysis value ₍ as _C84H122N4・_4HCl・3/ _2H2O ) Calculated value Actual value C (%) 74.14 74.32 H (%) 9.55 9.65 N (%) 4.12 4.11 Production example 3 N-geranyl-N,N',N'',N-tetraethyltriethylenetetramine Add 20 g of geranyl bromide to 200 ml of a chloroform solution containing 60 g of triethylenetetramine. 100 ml of a chloroform solution containing the mixture was added dropwise with stirring at room temperature over 1 hour, and the mixture was further stirred at room temperature for 3 hours.
After the reaction is completed, chloroform is concentrated from the reaction solution under reduced pressure, and then the concentrate is extracted with ethyl acetate. The extract is washed with a 10% aqueous sodium hydroxide solution and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 21 g of a concentrate. benzene concentrate
30 ml of acetic anhydride and 10 g of sodium acetate are added thereto, and heated under reflux for 4 hours while stirring. Convert the reaction solution into 300ml of 10% sodium hydroxide aqueous solution.
and extract with ethyl acetate. The extract is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 23.5 g of the concentrate was chromatographed on a chromatography column packed with 250 g of alumina using a mixture of ethanol and ethyl acetate as the eluent to obtain oily N-geranyl-N,N',N'',
N-tetraacetyltriethylenetetramine
Obtain 9.3g. Next, this N-geranyl-N,N′,
Dissolve 9.3 g of N'', N-tetraacetyltriethylenetetramine in 100 ml of anhydrous tetrahydrofuran, and add 5.0 g of lithium aluminum hydride little by little at room temperature while stirring. After stirring at room temperature for 1 hour, stir for an additional 3 hours. After cooling the reaction solution, 5 ml of a 20% aqueous sodium hydroxide solution is added, and after decomposition, insoluble materials are separated and the solution is concentrated under reduced pressure.The concentrate is extracted with isopropyl ether, and water and reflux are added. After washing with saturated brine, drying over anhydrous sodium sulfate, and concentrating under reduced pressure, the formula N-geranyl-N, N′, N″, N
- 5.1 g of tetraethyltriethylenetetramine are obtained as an oil. Next, the physical properties of this material are as follows. n ²² _D = 1.4851 NMR (δ value in CDCl ₃ ) 5.0 ~ 5.4 (2H, m) 3.10 (2H, d J = 7Hz) 0.8 ~ 2.9 (48H, m) Elemental analysis value (C ₂₄ H ₅₀ N ₄・H ₂ O) Calculated value Actual value C (%) 69.85 69.98 H (%) 12.70 12.81 N (%) 13.58 13.35 Production example 4 N-phytyl-N,N',N'',N-tetraethyltriethylenetetramine Triethylenetetramine 100 ml of a chloroform solution containing 31 g of phytyl bromide is added dropwise to 200 ml of a chloroform solution containing 60 g of phytyl bromide at room temperature over 1 hour with stirring, and the mixture is further stirred at room temperature for 3 hours.After the dropwise addition is complete, chloroform is removed from the reaction solution under reduced pressure. and the concentrate is extracted with ethyl acetate. The extract is washed with 10% aqueous sodium hydroxide solution and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 30 g of concentrate. Then concentrated. The product is dissolved in 100 ml of benzene, 30 ml of acetic anhydride and 10 g of sodium acetate are added thereto, and the resulting mixture is heated under reflux for 4 hours with stirring. After the reaction is complete, the reaction solution is dissolved in 300 ml of 10% aqueous sodium hydroxide solution. Open and extract with ethyl acetate. The extract is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 41 g of the concentrate is poured onto a chromatography column packed with 250 g of alumina using a mixture of ethanol and ethyl acetate. Chromatographed oily N
- 11 g of phytyl N, N', N'', N-tetraacetyltriethylenetetramine are obtained. Next, this N
11 g of -phytyl-N,N',N'',N-tetraacetyltriethylenetetramine are dissolved in 100 ml of anhydrous tetrahydrofuran, and 5.0 g of lithium aluminum hydride is added little by little at room temperature while stirring.
The mixture is stirred at room temperature for 1 hour and heated to reflux with stirring for an additional 3 hours. After cooling the reaction solution, 5 ml of a 20% aqueous sodium hydroxide solution is added to it, and after decomposition, insoluble materials are separated, and the resulting solution is concentrated under reduced pressure. The concentrate was extracted with isopropyl ether, washed with water and saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure.
formula N-phytyl-N, N′, N″, N
7.3 g of tetraethyltriethylenetetramine are obtained as an oil. Next, the physical properties of this material are as follows. n ²² _D = 1.4721 NMR (δ value in CDCl ₃ ) 5.30 (1H, t J = 7Hz) 3.05 (2H, d J = 7Hz) 2.0 ~ 2.9 (20H, m) 0.8 ~ 2.0 (49H, m) Elemental analysis value (as C ₃₄ H ₇₂ N ₄ · H ₂ O) Calculated value Actual value C (%) 73.58 73.78 H (%) 13.44 13.51 N (%) 10.10 7.89 Production example 5 N-decaprenyldiethylenetriamine Same as production example 1 By reacting decaprenyl bromide and diethylenetriamine, the formula N-decaprenyldiethylenetriamine represented by: is obtained. The physical properties of this product are shown in Table 1. Production Example 6 N-Solanesyltriethylenetetramine 4-hydrochloride Solanesyl bromide and triethylenetetramine were reacted in the same manner as in Production Example 1 to obtain the formula N-solanesyltriethylenetetramine tetrahydrochloride represented by is obtained. The physical properties of this product are shown in Table 1. Production Example 7 N-decaprenyldipropylenetriamine trihydrochloride Decaprenyl bromide and dipropylenetriamine were reacted in the same manner as in Production Example 1 to obtain the formula N-decaprenyldipropylenetriamine trihydrochloride represented by is obtained. The physical properties of this material are the first
As shown in the table. In the chemical structural formula in the table below, D represents a decaprenyl group, S represents a solanesyl group, and Phys.
indicates a phytyl group.

[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 0.1 ml of a diluted solution of vaccinia virus was intravenously injected 2 cm from the base of the tail to 10 ICR female mice weighing around 15 g, and 8 days after inoculation. Lesions that appeared on the surface of the tail were stained with a 1% fluorescein-0.5% methylene blue mixture and counted. The test compound was made into a suspension using a surfactant and administered intraperitoneally at 50 mg/Kg 24 hours before virus inoculation, and the antiviral effect was evaluated based on the lesion inhibition rate compared to the group administered only with the surfactant.
Table 2 shows the inhibition rate of each sample compound.

[Table] (2) Antitumor effect Balb/c male mice weighing around 20g, group 16
5 x ¹⁰⁵ tumor cells _KN7-8 in each animal
was administered intraperitoneally. The test compound was made into a suspension using a surfactant, and was administered three times, 24 hours before tumor cell transplantation, and 2 and 5 days after transplantation.
It was administered intraperitoneally at a dose of mg/Kg, and the antitumor effect was determined by the number of survivors on the 30th day after transplantation. Table 3 shows the number of survivors for each test compound.

[Table] (3) Human interferon-inducing effect (in vitro) A suspension of the test compound (after making it into an ethanol solution) was applied to human-derived normal diploid cells (fibroblast-like cells).
diluted with PBS (-), 25 nmol concentration).
It was induced according to the method of Edward A. Havell et al. The interferon induced inhibition rate of ³ H-uridine uptake was measured using the radioisotope microassay method of H. Ishitsuka et al. Table 4 shows the ³ H-uridine uptake inhibition rate of each test compound.

【table】

[Table] (4) Anti-vaccinia virus activity (in vitro) A suspension of the test compound (made into an ethanol solution and then suspended in Hanks culture medium to a concentration of 50 nmol) and virus dilution were applied to Vero cells derived from African green monkey kidney. The inhibition rate of virus plaque formation was determined by applying the solution. Table 5 shows the inhibition rates of the test compounds.

[Table] (5) Toxicity The 50% lethal dose was determined by intravenous administration using 20 to 25 g of ddY male mice. The result is the 6th
Shown in the 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 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 due to interferon alone but also to other host-mediated factors. It is also possible that sexual defense mechanisms are 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 route 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 gum arabic, gelatin, tragacanth, hydroxypropylcellulose or polyvinylpyrrolidone, lubricants such as magnesium stearate, talc or silica, disintegrants such as potato starch, carboxylic It may contain methylcellulose calcium, 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 fluid formulations 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 Oral hard capsule N-decaprenyltriethylenetetramine 25g
and 7.5 g of polyoxyethylene castor oil are dissolved in acetone, and then 25 g of silicic anhydride are mixed.
After evaporating the acetone, add 5 g of carboxymethyl cellulose calcium and 5 g of corn starch.
g, 7.5 g of hydroxypropylcellulose and 20 g of microcrystalline cellulose are mixed, mixed with 30 ml of water and granulated. This was granulated using a No. 24 mesh (BS) granulator (manufactured by Fuji Paudal Co., Ltd.) equipped with a screen.
The condylar flow was dried to less than 5% water content and sieved through a No. 16 mesh (BS) sieve. Next, the particles were filled to 190 mg per capsule using a capsule filling machine. Formulation example 2 Soft capsule for oral use N-decaprenyl-N,N',N'',N-tetrabenzyltriethylenetetramine tetrahydrochloride 50g
and polyethylene glycol (macrogol-
400) Mix 130g to make a homogeneous solution. Separately 93g gelatin, 19g glycerin, D-sorbitol
10g, ethyl paraoxybenzoate 0.4g, propyl paraoxybenzoate 0.2g and titanium oxide 0.4
A gelatin solution having the composition of 1.g was prepared, and this was used as a capsule coating agent to produce soft capsules containing 180 mg of content by manual plate punching. Formulation Example 3 Injection Mix 5 g of N-geranyl-N,N',N'',N-tetraethyltriethylenetetramine, an appropriate amount of peanut oil, and 1 g of benzyl alcohol, and use peanut oil to bring the total amount to 100 c.c. Dispense 1 c.c. of this solution into ampoules using aseptic technique and seal. Formulation example 4 Injection N-decaprenyltriethylenetetramine 1.0
g, Nikkol HCO60 [Nikkol HCO60 (product name)] (hydrogenated castor oil polyoxyethylene-60
mol-ether) 5.0g, propylene glycol
20g, glycerol 10g, ethyl alcohol 5.0
g, and add 100 ml of distilled water to this and stir. Dispense this solution into 1.4 ml ampoules using aseptic technique and melt and seal.

Claims (1)

[Claims] 1. General formula (where n is 2 to 10, A and B are each a hydrogen atom or together form a single bond, and n is 4
in which A and B may be a combination of the above two cases, R is hydrogen, benzyl or lower alkyl, p is 2 or 3 and q is at least 2) Prenylamine derivatives and acid addition salts thereof. 2 General formula (where n is 2 to 10, A and B are each a hydrogen atom or together form a single bond, and n is 4
in which A and B may be a combination of the above two cases, R is hydrogen, benzyl or lower alkyl, p is 2 or 3 and q is at least 2) An antiviral agent containing a prenylamine derivative and its acid addition salt as an active ingredient.