DE3043446C2 - - Google Patents

Description

The present invention relates to new decaprenylamine derivatives and their acid addition salts, which are useful for control of virus infections in vertebrates and humans.

There are various substances that a preventive or alleviating effects are attributed to diseases that caused by virus hosted by vertebrates or are believed to be capable of the symptoms of such diseases through noticeable intensification to alleviate antibody activity in animals. Known so far antivirals include interferon, substances that affect the Have the ability to induce interferon; H. Induction materials (Interferon induction substances), amantadine hydrochloride or synthetically produced substances such as methysazone, which immediate inhibitory effect on virus spread to have. Interferon is a glycoprotein, which possesses antiviral and antitumor activity. This glycoprotein is produced in situ by vertebrate cells when the cells are infected with virus. It has been recommended for the therapy of infectious viral diseases and also for cancer therapy. Known induction materials that Interferon in vertebrates using a method other than The virus infection induce natural high molecular weight Substances such as double chain ribonucleic acids from Bacteriophages of a certain species or synthetic high molecular weight Substances such as double chain ribonucleic acids. Typical examples of this are polyinosinic acid-polycytidylic acid, or low molecular weight induction materials like Tyrolon.  

However, there is a significant amount in the manufacture of interferon Problem in cleaning it, and so far there is none economic process for its production has been found. On the other hand, there are known interferon induction substances have not yet been used in practice, namely in essentially because of their toxicity. Synthetic antiviral Substances that have an inhibitory effect on the Exercise virus spread, and those currently commercially available have a very narrow range in terms of virus infected Diseases caused by administration of these Means are curable. Therefore there is a considerable need in the creation of new synthetic substances with antiviral Properties. The inventors of the present invention have done extensive research with the Aim to find such compounds that are used to produce Interferon with high effectiveness and beyond that antiviral activity at the biological level have. It has now surprisingly been found that the Compounds of the general formula I shown below and their acid addition salts an excellent interferon Induction ability and at the same time excellent antiviral activity even in the biological test demonstrate.

The present invention accordingly relates to a new one Class of decaprenylamine derivatives of the general below formula

wherein

n represents an integer from 0 to 2, R₁ represents a hydrogen atom, a lower alkyl group or a decaprenyl group, and R₂ represents a phenyl group or a pyridyl group

and its acid addition salts.  

Suitable lower alkyl groups, which R₁ may represent straight-chain and branched alkyl chains with up to 4 Carbon atoms.

For the preparation of the decrapenylamine derivatives of the above general Formula I and its acid addition salts can be a method are used in the known procedural measures for amine synthesis on the starting decaprenol following formula

can be used to produce a desired amine derivative.

The amine derivative obtained in this way can also be converted into a corresponding one Salt are transferred in a manner known per se. More specifically, a desired amine can be processed be prepared according to a suitable decaprenyl alcohol the above general formula II in a corresponding Halide or a sulfonic acid ester transferred and then with an appropriate primary or secondary Amine compound according to the desired end product in Presence or absence of a base is implemented. In an alternative The desired amine can be obtained by oxidation of a decaprenol to a corresponding aldehyde be, then with a corresponding primary amino compound is condensed, water being split off in order to create an appropriate imino compound, which in turn with a suitable reducing agent (e.g. sodium borohydride) is reduced. An acid addition salt from one so produced Amine derivative can be obtained in that the obtained Amine in a suitable solvent with an appropriate Acid mixed to form the salt and the salt crystallized or obtained by evaporation or otherwise  becomes. Examples of suitable addition salts for medical Application are e.g. B. salts with hydrochloric acid, acetic acid, Citric acid, fumaric acid, etc.

The preparation of the compounds I and their acid addition salts is explained using the following manufacturing examples:

Production Example 1 N-benzyl didecaprenylamine hydrochloride

To a solution of benzylamine (10 g) in ethanol (100 ml) was added dropwise at room temperature over 1 hour Stir a solution of decaprenyl bromide (20 g) in benzene (40 ml) and stirring was continued for a further 2 hours. The mixture obtained was then stirred for 2 hours on Heated to reflux. The reaction mixture obtained was after cooling with an aqueous 2N sodium hydroxide solution (100 ml) was added and then extracted with isopropyl ether. The liquid extract thus obtained was saturated with water and Washed saline, dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The Residue (21.4 g) was removed by column chromatography Cleaned using silica gel (200 g). The elution was performed with a mixture of isopropyl ether and benzene. The initially eluted fraction (7.0 g) was in ethyl acetate dissolved, mixed with HCl-containing ether to weakly acidic reaction and then cooled. The crystallized mass was filtered off, using N-benzyl-didecaprenylamine hydrochloride (4.9 g) F. 52 to 54 ° C was obtained. The elementary analysis for C₁₀₇H₁₆₉N · HCl gave the following values:  

calculated: C 85.34, H 11.38, N 0.93%; found: C 85.08, H 11.23, N 0.94%.

Production Example 2 N-benzyldecaprenylamine hydrochloride

The fraction eluted last according to Preparation Example 1 (7.3 g) was dissolved in acetone and HCl-containing ether was added. The mixture was made in the same manner as in Example 1 described worked up, wherein N-benzyldecaprenylamine hydrochloride (5.8 g), mp 105-107 ° C. The elementary analysis for C₅₇H₈₉N · HCl gave the following values:

calculated: C 83.00, H 11.00, N 1.70%; found: C 82.82, H 10.87, N 1.65%.

Production Examples 3 to 9

The same procedure as described in Example 1 was applied to the reaction of decaprenyl bromide with a primary or secondary amine compound, to produce the following compounds. Their structural formula, empirical formula, melting point and values of the elementary analysis are summarized in Table I.

Production Examples 10 to 13

To a solution of 3-aminomethylpyridine (25 g) in ethanol (100 ml) was a solution of decaprenyl bromide (30 g) in Chloroform dropwise at room temperature with stirring inside Added 1 hour. Stirring was continued for 3 hours. The reaction mixture obtained was after cooling mixed with an aqueous 2N sodium hydroxide solution (100 ml) and extracted with isopropyl ether. The liquid obtained Extract was washed with water and saturated saline, dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The residue (21 g) was determined by column chromatography using Silica gel (200 g) cleaned. The elution was initially carried out with a mixture of 20% ethyl acetate and 80% hexane. The 3-didecaprenylaminomethylpyridine (about 1.38 g) was used as Obtain oily substance.

The above-mentioned column was further mixed eluted from 20% ethanol and 80% ethyl acetate. The faction (8.6 g) was recrystallized from acetone, whereby crystalline 3-decaprenylaminomethylpyridine (7.8 g) was obtained.

Using essentially the same procedures, as described above, 2- (2-decaprenylamino) - ethyl pyridine, 2- (2-didecaprenylamino) ethyl pyridine and 3- (N- Decaprenyl-N-methylamino) -methylpyridine produced.

The physicochemical properties of these compounds are summarized in Table 2.

Production Example 14

To a solution of N, N-dimethylbenzylamine (10 g) in ethanol (100 ml) decaprenyl bromide (20 g) was added dropwise during Given for 30 minutes at room temperature. Then 1.5 Stirred for hours at room temperature. The reaction mixture obtained was extracted with isopropyl ether, and the extracted Phase was washed with water, over anhydrous sodium sulfate dried and concentrated under reduced pressure. The residue obtained (22.0 g) was purified by column chromatography cleaned over silica gel. Then through the column a mixture of ethyl acetate / chloroform with a concentration gradient given from 1 to 10% ethyl acetate. The eluted Fraction (18.6 g) was dissolved in acetone and the resulting one Solution was left in a refrigerator overnight, crystalline N-benzyl-decaprenylammonium bromide (15.8 g), F. 51.2 to 53.1 ° C to win.  

The physiological effects of the compounds of the present Invention are explained below:

1. Interferon induction activity

The test compound was surface-active Medium suspended in water groups of 5 female each ICR mice weighing approximately 25 g intraperitoneally administered. 20 hours after the administration, the blood was the mice were collected and the serum was separated to make a To obtain serum interferon. The ones described below Steps were performed in the order given to determine the potency of the serum interferon so induced. L-929 cells derived from mice and previously in one Monolayer had been incubated at 10-fold diluted test serum solution contacted overnight at Incubated 37 ° C in an incubator in a carbon dioxide atmosphere and the diluted test serum solution was removed therefrom. After that the cells were inoculated with vesicular stomatitis virus and placed on a tissue culture medium containing 1% agar. After 24 hours of incubation at 37 ° C, the cells were added a neutral red solution and to a suitable concentration diluted to count the number of plaques formed and thereby plaque inhibition rate in the respective test group to charge against a group that has no test compound had been administered. In Table 3 they are Plaque inhibition rates of the test compounds are compiled.

2. Effect on mice infected with vaccinia virus

Groups of 10 female ICR mice each were identified by the Tail vein vaccinia virus (DIE strain) injected intravenously. On the 8th day after the injection, the number of injuries in the form of small pox counted on the surface of the tail Staining the tail with an ethanol solution containing 1% fluorescein and 0.5% methylene blue. In this attempt  were the test compounds the day before the Inoculation of the virus administered intraperitoneally to the mice. The antiviral activity of the test compounds was expressed by that the inhibition of tail damage to the particular Experimental group was compared to those the group to which no test compound was administered was.

Table 3 shows the inhibition rates with respect to Tail lesions of the test compounds compiled.

3. Effect on mice infected with influenza virus

Groups of 10 female ICR mice each weighing of about 25 g was inhaled by inhaling nebulized influenza. Virus A / PR-8 infected. A solution of the respective test connection in an aqueous solution containing a surfactant Mean was given 24 hours and 3 hours before the virus infection and five times every other day after intraperitoneally the mice second day after infection administered. Those mice 21 days after infection survived were considered survivors, and the Survival rate was calculated according to the following equation:

Table 3

4. Toxicity

To determine the acute toxicity of the compounds according to the invention became the 50% lethal dose of the compounds on male ddY mice weighing 20 to 25 g determined. From the results summarized in Table 4 it can be seen that the connections have a high security area when administered intraperitoneally.

Table 4

From the test results above it is clearly evident that the active ingredients of the present invention in vivo have an interferon induction activity and are not very toxic and have excellent antiviral activity. In view of the fact that a strict correlation of the Interferon activity with the respective antiviral activity not always observed with the compounds according to the invention the possibility that the antiviral activity of these compounds in the biological field not only based on interferon, but also on one other host defense mechanism. If so  the active ingredients of the present invention used in the treatment of diseases infected by virus they can pass the patient through any Methods are administered, e.g. B. orally, by inhalation or by other administration methods and also by subcutaneous, intramuscular or intravenous injection. Depending on the condition the patient, e.g. B. its age, symptom and route of administration of the active ingredients becomes the active ingredient according to the Invention expediently in a dose of 0.5 to 20 mg / kg, preferably 3 to 5 mg / kg administered several times (2 to 4 times) daily.

The compounds according to the invention can be used in suitable preparations formulated for medical administration be, e.g. B. to tablets, capsules, granules, powder, liquid Preparations for oral use, eye drops, suppositories, Ointments, solutions for injection, etc.

When the compounds according to the invention administered orally they can be in the form of tablets, capsules, granules or powder. These solid preparations for Oral administration can use commonly used excipients included, e.g. B. silicic acid anhydride, metasilicic acid, Magnesium alginate, synthetic aluminum silicate, lactose, Cane sugar, corn starch, microcrystalline cellulose, hydroxypropylated Starch or glycerin, etc .; Binders such as rubber arabic, gelatin, tragacanth, hydroxypropyl cellulose or Polyvinyl pyrrolidone; Lubricants such as magnesium stearate, lime or silica, disintegrants such as potato starch and carboxymethyl cellulose; Wetting agents such as polyethylene glycol, Sorbitan monooleate, hydrogenated castor oil, sodium lauryl sulfate. For the production of soft capsules, the compounds according to formulated the present invention in particular that they are used in the usual way oily substrates like sesame oil, peanut oil, germ oil, fractionated coconut oil like Miglyol etc. can be dissolved or suspended. Tablets or granulate preparations can be coated according to customary methods will.  

Liquid preparations for oral administration can be in the form of aqueous or oily emulsions or syrups.

Alternatively, they can be in the form of dry products are present before use by means of suitable Carriers can be dissolved again. This liquid Preparations can be added to commonly used additives such as emulsifying aids such as sorbitol syrup, Methyl cellulose, gelatin, hydroxyethyl cellulose, etc. Emulsifiers such as lecithin, sorbitan monooleate, hydrogenated Castor oil, non-aqueous carriers such as fractionated coconut oil, Almond oil, peanut oil, etc .; or antiseptics like methyl p-hydroxybenzoate, propyl p-hydroxy benzoate or sorbic acid. These oral preparations can also if necessary, preservatives, stabilizers and the like Contain additives.

If the compounds according to the invention are in the form of non-oral Suppositories can be administered in known per se To be formulated using olephile substrates like cocoa oil, or them can be used in the form of rectal capsules that get are made by enveloping a mixture of polyethylene glycol, Sesame oil, germ oil, fractionated coconut oil etc. in a sheet of gelatin. The rectal capsules can, if desired, be coated with waxy substances.

If the compounds according to the invention as injection solutions used, they can be emulsified in oil solutions Solutions or aqueous solutions are formulated, and these solutions can contain commonly used emulsifiers, Contain stabilizers or similar additives.

Depending on the type of administration, the described Preparations the compounds according to the invention in one Amount of z. B. contain at least 1%, preferably 5 to 50%.  

Process for formulating the compounds according to the invention into different preparations are based below of the pharmaceutical examples explained.

Pharmaceutical Example 1 Hard capsules for oral administration

A mixture of 25 g of N-benzyldecaprenylamine hydrochloride and 7.5 g of polyoxyethylene castor oil in acetone was mixed with 25 g of silica mixed. After the acetone was evaporated the mixture further with 5 g calcium carboxymethyl cellulose, 5 g corn starch, 7.5 g hydroxypropyl cellulose and 20 g microcrystalline Cellulose mixed and 30 ml of water was given, and the mixture was then produced to produce a Kneaded granules. The mass was pelletized, whereby a pelletizing machine (ECK pelletter from Fuji Paudal Co., Japan) was used with a No. 24 mesh (B.S.) sieve was equipped. In this way, granules were produced. The granules were brought down to a moisture content of less than 5% dried and through a No. 16 mesh (B. S.) Sieve sieved. The sieved granulate was in a capsule filling machine filled into capsules, each containing 190 mg.

Pharmaceutical Example 2 Soft capsules for oral administration

By mixing 50 g of N- (methyl-N-benzyl) decaprenylamine hydrochloride with 130 g of polyethylene glycol (Macrogol 400) homogeneous solution. A gelatin solution was separated prepared, the 93 g gelatin, 19 g glycerin, 10 g D-sorbitol, 0.4 g ethyl p-hydroxybenzoate, 0.2 g propyl p- contained hydroxybenzoate and 0.4 g of titanium oxide, and which as Capsule film-forming agent was used. The first one received Solution was made together with the capsule film-forming agent treated in a manual capsule forming machine, whereby Capsules were obtained, each containing 180 mg.  

Pharmaceutical Example 3 Solution for injection

A mixture of 5 g of N-methyl-N-phenyl-decaprenylamine hydrochloride, an appropriate amount of peanut oil and 1 g of benzyl alcohol were made up by adding peanut oil to a total volume brought from 100 ml. The solution was poured into ampoules in portions, containing 1 ml each, filled under aseptic conditions. The ampoules were then sealed.

Pharmaceutical Example 4 Solution for injection

A mixture of 1.0 g of N-phenethyl-decaprenylamine hydrochloride, 5.0 g Nikkol RCO-60 (trademark) (hydrogenated castor oil / polyethylene 60 mol ether), 20 g propylene glycol, 10 g glycerol and 5.0 ethyl alcohol was mixed with 100 ml of distilled water and stirred. This was done under aseptic conditions Solution in portions in amounts of 1.4 ml in ampoules bottled, which were then melted.

Claims (6)

1. Decaprenylamine derivatives of the formula: wherein n represents an integer from 0 to 2, R₁ represents a hydrogen atom, a lower alkyl group or a decaprenyl group, and R₂ represents a phenyl group or a pyridyl group and their acid addition salts. 2. N-Benzyldecaprenylamine hydrochloride. 3. N-Benzyldidecaprenylamine hydrochloride. 4. 3-decaprenylaminomethylpyridine. 5. 2-decaprenylaminoethyl pyridine. 6. Use of compounds according to claim 1 in the Fighting virus infections.