DE2760410C2 - - Google Patents

Description

The present invention relates to the use of verbenon in the treatment of diseases of the respiratory system.

Swiss patent 5 42 163 and the corresponding BE-PS 7 84 885 describe a process for the production of terpene fractions which are suitable for the treatment of bronchopneumonic diseases. In this process, mixtures are primarily oxidized that contain α- pinene. According to this process, in particular two terpene fractions are obtained which distill within a temperature range from 40 ° C to 60 ° C or 65 ° C.

The above oxidation mixtures can undergo additional treatment get abandoned. For the production of verbenon this treatment in another, with chromium trioxide Oxidation carried out in sulfuric acid. It shows that if you look at the oxidation mixture of the Swiss Patent document treated with chromium trioxide in sulfuric acid, the oxidation to a complete disappearance of the alcohols and for the formation of carbonyl compounds (verbenone and Myrtenal) leads. The treatment with chromium trioxide Ketones obtained in sulfuric acid can be treated with a bisulfite Complex and isolated by fractional distillation.

For what therapeutic purposes the compounds so produced can be used follows from the known usability of the terpene fractions of the Swiss Patent 5 42 163 and in particular the parliamentary group, which distills between 65 ° C and 105 ° C (hereinafter referred to as "Fraction 2").

Fraction 2 shows both a soothing and an analeptic Effect and is used for appropriate therapeutic Purposes.  

Apart from the fact that the active substances are scientific do not identify exactly on which the therapeutic Effectiveness of pharmaceutical, on this fraction 2 based preparations, it is in technical terms to be regarded as a disadvantage that a mixture of compounds must be processed, their connection - albeit within a certain range - is changeable. It it must also be noted that pharmaceuticals that Preparations containing fraction 2 in addition to the linden tree mentioned properties as a general indication in less Measurements of some other effects show that it is not it is possible to have one of these effects stronger than the other to emphasize or to improve the general effect.

It has now been found that that in the above fraction 2 in Traces of existing verbenon with the structural formula

in addition to the properties already mentioned, such as. B. the soothing and analeptic effect, also clearly bronchodilatory, anti-inflammatory, anti-exudative and anti-aggregative Shows effects.

The invention is accordingly the in claim defined use of verbenon.

To demonstrate the pharmacological effectiveness of verbenone, the following parameters were used:

• 1. Acute toxicity
• 2. Effect on the bronchial muscles
• 3. Influence on inflammatory processes
• 4. Effect on hemolysis
• 5. Effect on platelet aggregation
• 6. Antibacterial effect

In all trials, the effectiveness of Verbenon was assessed that of Fraction 2 described above and pharmaceutical Preparations based on terpene alcohols, in particular Sobrerol, compared.

1. Acute toxicity

Table I below shows the LD₅₀ of verbenone (and the confidence limits 95%), which according to the Litchfield procedure and Wilcoxon (J. Pharmacol. Exp. Ther., 96, p. 99, 1949) was determined. Each dose of verbenon was intraperitoneal (i. p.) and orally (p. o.) a group of 10 mice or 6 rabbits administered.

Table I

Because of these values, Verbenon appears to be a well tolerated compound to be if it is these animal species in the manner mentioned  is administered because the Gleason rating is only moderate to low toxicity.

2. Bronchodilatory effectiveness

Verbenon shows a pronounced, both in vitro and in vivo bronchodilator effect and differs in a very clear way from the other medically effective Substances of fraction 2 of Swiss patent 5 42 163 and the terpene used for comparison, i.e. H. Sobrerol.

a-in vitro

In a guinea pig trachea, which had been isolated according to the method of Costantine (J. Pharm. Pharmcol., 17, page 384, 1965), verbenone in a concentration of 125 to 2000 μg / ml caused the smooth tracheal muscles to relax was significantly higher than in the case of the terpenes of fraction 2 and the sobrerol (see Table II). In concentrations from 10 ^-3 M to 5 · 10 ^-3 M it inhibited the contractions caused by histamine of the isolated guinea pig trachea (histamine 10 ^-5 M, see Table III).

Table II

Percent relaxation (average ± standard deviation of 4 preparations) of the isolated guinea pig trachea at concentrations of 125, 500 and 2000 µg / ml

Table III

Percentage inhibition achieved with verbenon of the histamine spasm caused by slackening of the smooth tracheal muscles (average + standard deviation of 4 preparations)

b-in vivo

Verbenon, the anesthetized dogs in doses from 0.6 to 4.8 mg / kg body weight injected intravenously resulted in a significant reduction in lung resistance (see Table IV), as in the Diamond (Arch. Int. Pharmacodyn., 168, page 239, 1967). Also inhibited Verbenon when it is rabbits by intravenous perfusion in dosage units of 2.5 ml / kg body weight per minute was a bronchospasm experimentally induced by histamine (100 µg histamine per body weight, intravenously) essentially to a greater extent than the terpenes of fraction 2 and as sobrerol (see table V).

Table IV

Percentage reduction in dog lung resistance when administered intravenously (average ± standard deviation from 4 trials)

Table V

Percentage inhibition of histamine-induced bronchospasm in rabbits with intravenous perfusion (average ± standard deviation from 4 experiments

3. Anti-inflammatory effect

Albino rats of the COBS breeding strain (Charles River) intraperitoneally a dose of 30 mg test compound per kg body weight, one was experimented with carrageenin edema produced (Winter et al., Proc. Soc. Exp. Biol. Med., 111, 544, 1962) on the paw both in normal rats as well in rats with the adrenal gland removed Use of verbenon inhibited much more than with the Fraction 2 and sobrerol (see Table VI).

In doses of 36 and 120 mg / kg body weight intraperitoneally administered verbenon shows in an experimental Turpentine-induced pleurisy (Hurley et al, J. Path., 91, Page 575, 1966) a similar anti-exudative effect as Aspirin (see Table VII).  

Table VI

Anti-inflammatory effect on carrageenin-induced edema in rats

Table VII

Effect on turpentine-induced pleurisy in rats

4. Anti-hemolytic activity, in vitro

With an effective concentration of 50% (EC 50) of 639.5 µg / kg Body weight (confidence limits 95%: 518.0-760.11) protects verbenon in vitro the red blood cells of rats in front of one due to capillary activity Medium ("Tween®80") induced hemolysis.  

5. Antiaggregative activity, in vitro

Verbenon inhibits at concentrations of 160 to 1280 µg / ml platelet aggregation of ADP in vitro (evaluated according to the method of Born and Cross, J. Physiol., London, 168, page 178, 1963) to a greater extent than the terpenes of fraction 2 and of sobrerol (see Table VIII).

Table VIII

Percent inhibition of platelet aggregation (average of several attempts)

6. Antibacterial effectiveness

Verbenon shows only a slight antibacterial effect on gram- positive and gram-negative germs with the least inhibition ration concentration (MIC) of 800 µg / ml in Staphylococcus aureus and Escherichia coli (see Table IX), but is also effective like fraction 2 and more effective than sobrerol.

Table IX

MIC in µg / ml

In summary, it can be said that Verbenon - in one strong contrast to fraction 2 of the Swiss patent 5 42 163 and on Sobrerol - a distinct bronchodilator Shows effect.

It can therefore be used in particular as an active ingredient in pharmaceutical preparations for the treatment of bronchopneumonic diseases where the respiratory tract through Inflammation or clogged infections are used. For therapeutic A dose of 10 to 100 mg verbenone per dose is recommended for purposes Day.  

The verbenon used according to the invention can in a known manner to pharmaceutical preparations to be administered orally of normal effect or long-term effect, e.g. B. softer in shape Capsules, or to injection liquids, suppositories, Sprays processed in various solution forms, ointments and creams be used in the pharmaceutical industry usual carriers and fillers can be used. Verbenon can either be the only active ingredient in the preparation form or in association with appropriate drugs, d. H. Antibiotics, antibacterial substances, chemotherapeutic Substances, sulfonamide preparations, anti-inflammatory agents, Cortisone preparations or analgesics.

The production of verbenon for the use according to the invention can be done as follows:

example (a) Alcohol oxidation

100 g of the terpene mixture were added to 2000 ml of dry acetone solved according to the Swiss patent specification 5 42 163 was obtained as fraction 2; this mixture consisted of 20 up to 30% from compounds with one carbonyl part of the molecule (predominantly Verbenon and Myrtenal) and 50 to 60% from compounds with an alcoholic part of the molecule (mainly verbenol and myrtenol).

The oxidation solution was prepared separately by carefully 37.41 g of CrO₃ and 32.2 ml of concentrated H₂SO₄ in of such an amount of water that the final volume of the Solution was exactly 140 ml.

The chromic acid was now slowly and dropwise into the acetone solution stirred while cooling on an ice bath to keep the temperature below 30 ° C. After completion The addition was made using commercially available diatomaceous earth  filtered and the filtrate worked up by adding acetone evaporated under reduced pressure at 40 ° C. The residue was diluted with water, cold neutralized with 10% NaOH and extracted with CHCl₃ until the mother liquor is exhausted. The combined organic extracts were over anhydrous Na₂SO₄ dried and by evaporation under reduced Pressure dried at 40 ° C.

In this way, about 100 g of a crude product were obtained, 50 to 60% from compounds with a carbonyl Part of the molecule (mainly Verbenon and Myrtenal) existed.

(b) separation of the carbonyl compounds from that by the Oxidation obtained raw material

A suitable vessel was charged with 100 g of that obtained in step (a) raw mixture, dissolved in 500 ml of ether, and with a solution of 120 g NaHSO₃ and 72 g NaHCO₃ in 2000 ml of water. The mixture was at room temperature for about 20 hours shaken vigorously and then poured into a separatory funnel; the organic phase was discarded. The aqueous phase was washed twice with 500 ml of ether, whereupon the obtained aqueous bisulfite solution and a water was subjected to steam distillation. The processing of the De stillats gave 50 to 55 g of a mixture that was practically complete consisted of pure verbenon and myrtle.

(c) Obtaining pure verbenon

50 g of the product from stage (b) were removed in vacuo at 2.7 kPa fractionated, with a back filled with nickel chips flow column was used. The distillation became slow performed (5 to 8 ml / h) by throughout the process step maintain a reflux ratio of about 40: 1 has been. The fraction distilling at 90 ° to 95 ° C consisted of myrtenal (19 g).  

The distilling at 110 ° to 113 ° C and 2.7 kPa Fraction consisted of verbenon (28.5 g) and had the following characteristics values:

n _D ¹⁰ = 1.49, d ₂₀ = 0.97 g / cm³
IR analysis: bands at 1670 cm ^-1 γ (C = O conjugated), 1615 cm ^-1 γ (C = C conjugated), 1650, 1435, 1370 cm ^-1
UV analysis; λ _max (C₂H₅OH) = 250 nm, ε = 7300
Semicarbazone: F = 188-190 ° C

Claims (1)

Use of verbenon in the treatment of diseases of the respiratory system in dosage units with one Verbenon content from 10 to 100 mg.