LU80739A1 - MEDICINAL PRODUCTS WITH EFFECT AS PROSTAGLANDIN SYNTHETASE INHIBITOR - Google Patents

Description

* * m

Medicinal products that act as prostaglandin synthetase inhibitors.

The invention relates to medicaments which act as a prostaglandin synthetase inhibitor *

Prostaglandins are common in all mammalian organisms. For a few years now, research has focused intensely on the isolation and knowledge of biological

The importance of prostaglandins. According to current knowledge, there are numerous prostaglandins with slightly different structures, their biological significance; in their wide distribution, their high effectiveness and the striking breadth and variety of their metabolic interactions. These different effects are due to the fact that intracellular prostaglandin synthesis can be triggered by an irritation or damage to the cell membranes, in the first phase phospholipases from membrane lipids prostaglandin synthesis precursors that different hormones such as e.g. Bradykiniri? Acetylcholine histamine increases the synthesis and release of prostaglandins and that prostaglandins stimulate both the adenyl cyclase system and the guanyl cyclase system and can thereby lead to an increase in the intracellular] APM and GJ-M concentrations.

However, as has been shown experimentally, the prostaglandin effects vary depending on the prostaglandin types used and the organs examined. For example, the adenyl-cyclase in endocrine organs> is stimulated by prostaglandins E1 and E2, but is inhibited in adipose * * tissue. This explains why

Prostaglandins, for example, can both increase and decrease the AMP level of the cell in the successful organ and develop an adrenaline - 2 - - 2 - and glucagon-antagonistic effect in adipose tissue. On the smooth muscles, prostaglandins sometimes cause contractions, such as in the uterus and intestines, but sometimes also dilate, such as in the vessels. In the kidney, for example, prostaglandins and Ag increase sodium and potassium excretion.

In addition, it is already known that an increase in tissue levels of prostaglandin Eg and inflammatory

Initiates and maintains reactions.

* * Numerous therapeutic applications result from the diverse metabolic effects of prostaglandins. For example, prostaglandins are used in the treatment of asthma and circulatory diseases, since prostaglandins of the E type show a vasodelating activity.

On the other hand, prostaglandins trigger contractions and initiate childbirth so that they can also be used as abortives if necessary.

It has only recently become known that the action of some drugs, some of which have been known for decades, with analgesic and anti-inflammatory activity, is based on an inhibition of prostaglandin sythetases. This applies, for example, to acetosalosalicylic acid, indomethacin and ibuprofen. The strong inhibitory effect of this compound explains on the one hand its effectiveness against inflammatory phenomena, but on the other hand also the occurrence of numerous side effects, of which only the triggering of gastric bleeding is mentioned as an example.

The biosynthesis of prostane glandins starts from the membrane phosphorus lipids, which are converted into arachidonic acid and converted to endoperoxide prostaglandins by oxygen radicals. The relatively stable prostaglandins, thromboxanes - 3 - - 3 - I 4 and the relatively unstable prostacyclin are formed from these endoperoxide prostaglandins.

The formation of PGE ^ unt * PGFg ^ aua arachidonic acid can be represented as follows: ά NSSNNNnHH ou „, // A \ /// NN: oo ft ον,

The biological half-life of the prostaglandins and especially the prostaglandin precursors is very short. The degradation takes place rapidly by oxidation on the C atom 15 and ♦ then via the p-oxidation typical of the fatty acids ·

It is already known that certain chemical compounds are strong prostaglandin synthetase inhibitors.

These compounds, such as indomethacin or acetylosalicylic acid, are used as inhibitors for the - h -

PfiEg-Syniietaae viewed and used accordingly for the treatment of inflammatory symptoms of various origins such as rheumatic and arthritic diseases and the like. The pronounced inhibitor effect, which does not only appear to be restricted to PGEg synthetase, also leads to the undesirable side effects caused by this, such as triggering gastric and intestinal bleeding, other diffuse bleeding, the occurrence of allergies and the possibility of influencing pregnancy .

• The invention is therefore based on the object of developing a new> drug with action as a prostaglandin synthetase inhibitor which does not have the known disadvantages.

To achieve the object, a medicinal product having an effect as a prostaglandin synthetase inhibitor is proposed, which is characterized in that it contains sterol glylcosides and / or their esters and / or spiroketal steroid glycosides and / or their esters as the active substance.

Completely surprisingly, it was found that sterol glycosides, spiroketal steroid glycosides and their esters are effective inhibitors for prostaglandin E2 and prostaglandin _synthetases and nevertheless do not trigger any side effects that otherwise occur as a result of influencing the prostaglandin level.

Sterolines occur quite frequently in nature in plants and microorganisms *, even if only in small quantities. Sterols are the glycosides of phytosterols including cholesterol and sterol-like tetracyclic triterpenes such as lanosterol and cycloartenol.

Some of these compounds are found in somewhat larger amounts in various plants, such as, for example, campesterol and stigmasterol and in particular sitosterol. The phytosterols correspond to the following general formula - 5 -

iA

in which R \ ^ and ^ are hydrogen atoms or methyl groups and in which ^ can be a hydrogen atom or a methyl, ethyl, methylene or ethylidene grouping. In addition, double bonds can be present at various points on the basic structure; this also applies to the silk chain. '

Most plants are at a certain level of phytosterols

Partly as sterol glycosides, that is, as sterolines and optionally as their esters. Most occurring in nature

Sterolines are monoglycosides, although some diglycosides have also been described. In addition to the D ^ -glucose most commonly found as sugar, which is usually linked to the 3-ß-hydroxyl group by an equatorial or ß-glycoside bond, / was found in the natural compounds, mannose, galactose, arabinose and xylose. In the naturally occurring esters, these have been identified as Bjteijfeinbasische1 monocarboxylic acids.

- 6 - and sisalagenin apply.

In nature, the agglomerates of saponins exist as glycosides and usually contain 3 or more Mongjça'cchàrideinfieiten in the sugar portion. For this reason, the relatively sugar-rich compounds are readily water-soluble and often form a soap-like foam.

The spiroketal steroid glycosides correspond to the following general formula u L ^ / 2.

in the 5-position there is a double bond or an alpha-hydrogen atom and in the Z is a mono- or bisaccharide and * in particular glucose, optionally esterified.

The sterolines and spiroketal steroid glycosides and their esters used according to the invention can be used as extracts from plant material, as enriched extracts, or as synthetically produced compounds.

The synthesis is carried out in a manner known per se, for example by the known Königs-Knorr synthesis for the production of glycosides using the corresponding aglycones, an ara C-1 brominated sugar acetate and silver oxide and silver carbonate.

When using sterolines and their esters, however, it should be noted that these have a high degree of insolubility in water. Sterolines must therefore be given in a particle size that is sufficiently small for absorption. It is therefore absolutely necessary that the sterolines used according to the invention are manufactured and / or prepared and / or incorporated into pharmaceutical preparations in such a way that liquid or solid solutions, emulsions or solid dispersions are formed which are known in a manner known per se by adsorption, absorption or by Milling processes with or without additives can be obtained. These methods all aim at reducing the size of the particles and reducing the crystallinity, so that they are present as minute amorphous mono- or multimolecular aggregates instead of in the form of crystalline microparticles. The compounds to be used according to the invention are mostly used with particle sizes of approximately 0.1 mm and preferably of 0.06 mm and smaller. The same applies despite the somewhat better solubility in water for the spiroketal steroid glycosides, which are also used with particle sizes of approximately 0.1 mm and preferably of 0.06 mm or smaller. 1

The compounds used according to the invention are administered in daily doses of about 0.03 to about 10 mg.

As a maintenance or prophylactic dose, approximately o. ^ 5 to o. L mg are usually given daily. These doses can be divided into three single doses or administered in a single dose with an extracted effect.

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To the best of our knowledge, the compounds to be used according to the invention are suitable for the treatment of those diseases in which a reduction in the PGE0 or the PGF level is required 6 ww 1. These are, for example, 1.) Ulcers, especially those of the gastrointestinal

Tract, 2.) en.docrine disorders, 3.) genitourinary disorders, especially benign

Prostate hyperthrophy and related complaints, 4.) heart diseases and blood pressure abnormalities, 5 ·) edematous conditions, 6.) vascular diseases, thromboses, varicose veins, and hemorrhoid pids, 7 ·) derraatitis and H1 · excess of stamines, 8.) inflammatory symptoms, • 9 ·) arthritic and rheumatic diseases, lo.) Allergies including asthma.

In a corresponding manner, the compounds to be used according to the invention can also be used for the treatment of animal diseases. The doses in the fight against - 9 - «t

Animal diseases can be calculated in a known manner, i.e. based on weight, assuming an average human weight of 75 kg.

The compounds can be processed into pharmaceutical specialties in a manner known per se wi®. for example to powders, pills and tablets, capsules, dragees, emulsions, solutions, solutions for injection or infusion, ointments and creams.

The invention is explained in more detail below with the aid of the examples:

Example 1;

Preparation of sitosterol (i-D-glucoside.

A mixture of 41.4 g of sitosterol and 55.2 g of silver carbonate in toluene is distilled with stirring until the distillate passes over anhydrous. A solution of 82.2 g of acetobromoglucose in 100 ml of toluene is then added dropwise to the stirred boiling mixture. The toluene is continuously distilled so that all of the water formed in the reaction is removed azeotropically.

The reaction vessel is protected from light during this time. If necessary, the volume of the reaction mixture is switched to constant by adding dry toluene. After the bromoacetoglucose solution has been added, the boiling is continued until the distillate is anhydrous. The reaction mixture is then filtered off and the residue is washed out with fresh hot toluene. The combined filtrates and washes are then evaporated under reduced pressure. The residue is recrystallized from ethanol or hexane. The yield of sitosterol glucoside tetraacetate is 22.4 g, corresponding to 30%.

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A solution of 4 g of sodium in 100 ml of ethanol is rapidly added with stirring to a solution of 10 g of sitosterol glucoside tetraacetate in 600 ml of ethanol at a temperature of 45 ° C. The mixture is stirred for one hour before 2 liters of water are added and the mixture is stirred for another hour. The precipitated sitosterol glucoside is filtered off and washed neutral with water before being dried in vacuo for 12 hours.

The yield is 6.9 g corresponding to 95%.

By choosing suitable starting compounds, all the other sterolines mentioned can also be obtained according to the *

Processes are prepared.

Example 2:

Preparation of diosgenin-3-jb'D-glucoside.

41.4 g of diosgenin and 55.2 g of silver carbonate were placed in boiling toluene, and the mixture was distilled with stirring until the distillate passed anhydrous. A solution of 82.2 g of bromoacetyl glucose in 100 ml of toluene was then added dropwise to the stirred boiling mixture. The mixture is continuously distilled to remove the water formed in the reaction. During this time, the reaction vessel is protected from light. If necessary, the volume of the reaction mixture is kept constant by adding dry toluene. After the addition of the acetrobromoglucose solution, the distillation continues until the distillate passes over anhydrous. The reaction mixture is then cooled and filtered. The residue is washed out with fresh hot toluene. The combined filtrates and washing liquids are screened to dryness under reduced pressure. The residue is recrystallized from ethanol or hexane. The yield of diosgenin-3-ft-D-glucoside tetraacetate was 25.5 g or 34.3%.

1 g of sodium was dissolved in 100 ml of absolute ethanol.

15 ml of this solution were quickly added with stirring to a solution of 10 g of diosgenin-glucoside-tetraacetate in 600 ml of ethanol at 45 ° C. Hour is stirred.

The precipitated diosgenin-β-D-glucoside is filtered off and washed neutral with water before it is dried in vacuo for 12 hours. The yield was 7 g or 90%. >

All other spiroketal steroid glycosides mentioned can also be prepared in a corresponding manner.

Example 3.

Manufacture of pharmaceutical preparations.

a) Production of lactose maize starch powders containing oiosgenin-ß-D-glxikosid.

15 g of diosgenin-ß-D-gluco3id are dissolved in 3 1 of a boiling mixture of chloroform and ethanol in a ratio of 3 * · 1. The solution is then added to 1 kg of lactose with a particle size of not more than 0.15 mm. The resulting slurry is evaporated to dryness with constant stirring.

The dry impregnated lactose is on the original. The particle size was crushed again and finally mixed with 9 kg of corn starch and 50 g of magnesium sterate. ·

This mixture is ideal for filling in capsules. Each capsule can contain, for example, 100 mg of the mixture, which corresponds to a content of 0.15 mg of diosginine, β-D-glul: 10 mg of lactose, 90 mg of corn starch and 0.5 mg of magnesium sterate.

b) Production of lactose granules containing diosgenin-ß-D-glucoside.

* I.

• · V *. Î.

Μ. * 1 " * ' .

- 12 "5 g dioagenin-ß-D-glucoside are boiling in 5 1> i,

Ethanol dissolved. The solution is then added to 3 »32 kg lactose with a particle size of not more than 0.15 mm. The slurry is evaporated to dryness with constant stirring. The dry impregnated lactose is comminuted to the original particle size before it is processed into granules with a preferred particle size of about 0.7 to 1.2. This granulated product is also particularly suitable for further processing in capsules, with for example one capsule containing 100 mg of the granules then containing 0.15 mg of diosgenin-β-D-glucoside. ·. ·

Products as under a) and b) can also be produced using i) glycosides of the 3-β-hydroxypiroketn steroid mentioned and in particular the β-D-glultosides from Tigogenin and Heco, genin.

ii) glucose, ascorbic acid or talc as carriers for the glycosides or also using other inert pharmaceutically acceptable carriers.

iii) The content of active glycoside compounds in each capsule can be set between o, ol mg and more.

iv) The auxiliary substances mentioned in a) and b) can be found in. Corresponding to the usual pharmaceutical manu factures 1.1 are changed.

v) Other pharmaceutically active compounds can be incorporated at any stage of the manufacturing processes in a) or].

c) Manufacture of tablets containing - 1% - *. J * · l, 25o g of diosgenin-β-D-glucoside are dissolved in 1 l of chloroform and mixed with 90o g of lactose. The slurry is dried under reduced pressure and constant stirring at room temperature. Then the mixture is mixed with 21oo g of potato starch and mixed vigorously again. The impregnated lactoso-starch mixture is mixed with 25oo ml of an aqueous solution of 25o g gelatin and 5 g glycerin and granulated in a manner known per se. The granules are dried under reduced pressure at room temperature.

The granulate is then> in a manner known per se.

pressed into tablets with a total weight of 4oo mg. Each tablet accordingly contains 0.15 mg of diosgenin-β-D-glucoside, 110.56 mg of lactose, 257 »97 mg of potato starch, 3 °, 3t mg of gelatin and o.61 mg of glycerin» d) Preparation of coated tablets containing Hecogenin-β-D-glucoside.

A solution of 45 ° mg of hecogenin-β-D-glncoside in 2 l of chloroform is mixed with 1050 g of lactose and 300 g of sucrose. The slurry is dried at 3 ° C. under reduced pressure and then granulated in a manner known per se after the addition of 1.6 l of an aqueous gelatin solution containing 40 g of gelatin. The granules are dried under reduced pressure and at a temperature of 45 ° C. Then the granules are intimately mixed with lo g of magnesium sterate »

The mixture thus produced (22oo g) is pressed to about 3ooo kernels, which are then coated in the usual coating process: *> with an optionally colored, thin Dragoec'ecko. Each coated tablet therefore contains 0.15 mg hecogenin-β-D-glucoside, 6l6.6? mg lactose, Ιοο, οο mg sucrose, 13.33 mg gelatin and 3 »33 mg magnesixtmsterate.

e) Preparation of an ointment containing

Hecogenin-β-D-glucoside.

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Cetylstearyl alcohol incorporated. After adding .105 g of viscous paraffin and 105 g of white petroleum jelly, the mixture is melted to 60 ° C. in a water bath. The melt is mixed with 699 g of water at approximately the same temperature in small proportions. The mixture is stirred until cool and gives an ointment containing 0.1% glucoside.

f) Preparation of a cream containing tigogenin (S -D-glucoside.

lg of tigogenin-D -D-glucoside is introduced into 500 g of wool wax alcohol and heated to about 50 ° C. in a water bath.

The mixture is then mixed with 499 g of water of approximately the same temperature in small proportions. The cream is stirred until it cools, whereby evaporated water components are added. The cream contains 0.1% glucoside.

In the same way as described in this example, the sterol glycosides to be used according to the invention can be processed into pharmaceutical specialties.

Example ^ L 4:

Manufacture of pharmaceutically acceptable solutions.

a) Solution containing semisynthetic soyasterol- | 2> -D-glucoside.

A solution of 10 g of polyvinylpyrrolidone in 4 l of distilled water at a temperature of 65 ° C. is added to a boiling solution of 600 mg of semisynthetic soyasterol-β-D-glucoside in 6 liters of absolute ethanol. The cooled 60% ethanol solution is filled into 250 ml bottles. Patients are instructed to take half a teaspoon equivalent to 2.5 ml of this mixture 3 times a day.

· - «- B -

The total solution gives 40 250 ml bottles, each containing about 100 2.5 ml cans and therefore sufficient for a treatment of about 33 days. Each 2.5 ml teaspoon contains 0.15 mg sterolines, 2.5 mg PVP and 1.5 ml ethanol.

It should be noted that concentrations of over 0.075 mg of sterolines and 1 mg of PVP per 100 ml of 60% aqueous ethanol should not be exceeded if clear solutions are desired, ie that about 0.1875 mg of sterolines per 2.5 ml aqueous 60% ethanol with a clear solution represent the maximum dose.

Solutions of other sterol monoglycosides or monoglucosides can also be prepared by the process described, but all of these compounds have low solubilities, so that these solutions should not be exposed to low temperatures by the described method for preventing turbidity.

b) Solutions containing semisynthetic sitosterol- (¾ -D-maltoside.

800 mg of sitosterol-P> -D-maltoside are dissolved in a mixture of 3 1 of ethanol and 7 1 of water at gentle reflux. The cooled 30% aqueous ethanol solution is then filled into 250 ml bottles.

Patients are instructed to take 2.5 ml of this solution 3 times a day (half a teaspoon or a full teaspoon, depending on the size).

* The total solution gives 40 250 ml bottles, each containing K 100 doses of 2.5 ml, so that the total amount is sufficient for a treatment of about 33 days. 2.3 ml of the solution contain 0.2 mg sterolines and 0.75 ml ethanol.

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Solutions of other sterol disaccharides can also be prepared by the process described. The

Water solubility of the ß-D-maltoside, the ß-D-lactoside and the ß-D-callobioside of sitosterol is 0.38 mg, 0.21 mg and 0.75 mg / 1 ml of water at a temperature of 24 ° C. These solubilities are above the preferred single doses of the compounds.

The preferred single dose for sterol disaccharides is 0.2 mg, i.e. 0.6 mg per day.

It is pointed out that, together with the sterol disaccharides, other pharmaceutically active compounds can also be incorporated into the solutions. In addition, the alcohol content of these sterol solutions can be changed; if appropriate, other pharmaceutically acceptable solvents can also be used. Pure water can also be used as the only solvent.

As described in this example, the spiroketal steroid glycosides can also be processed into pharmaceutically acceptable solutions.

Example 5:

Pharmacological testing of sterolines.

Toxicity testing of sterolines and spiroketal steroid glycosides.

When testing the acute toxicity in rats, mice, rabbits, dogs and primates, after oral administration of * e.g. Sitosterol-ß-D-glucoside even in doses of 1 to 2 g / kg * body weight no toxic effects were found.

»4 - 17 -

Even when administered over a longer period of daily doses of loo to 2oc mg / kg body weight, no toxic or gout-like symptoms could be found in these animal species, so that the tolerance can be described as good.

Example 6:

Proof of efficacy as a prostaglandin synthetase inhibitor.

The activity of the compounds as prostaglandinsynthetic inhibitor was determined by the procedures of A.L. Willis proven. Corresponding test conditions are described, for example, in Proceedings of a Workshop Held during the VIII th European Rheumatology Congress Helsinki 1975.

Siliconized KUVettes of the aggregometer are used as an incubation vessel at a temperature of 37 ° C, in which an arachidonate solution is quickly added and mixed with a prostaglandin synthetase enzyme system, usually made from sheep's bladder. Platelet-rich plasma treated with anticoagulants, which was also heated to 37 ° C., is then added to this solution in the cuvette. The light transmission through the cuvette is recorded immediately after addition. After 45 seconds of incubation, the control sample shows a clear peak in platelet aggregation, which shows the formation of prostaglandins PGE and PGFrt and the resulting platelet-à z,; k_ aggregation.

«4 -½ -

In the test samples with the addition of ο, οοοοί% sterol glycosides or spiroketal steroid glycosides, the 31utt platelets are not aggregated. This clearly indicates that the formation of prostaglandins via endoperoxide compounds from the arachidonate was inhibited.

Example 7t

According to newer knowledge, the prostaglandins PGE0 and PGF and the endoperoxide precursors of these compounds play an essential role in triggering rheumatoid arthritis. The effectiveness of the compounds to be used according to the invention as prostaglandir.synthetase inhibitors was therefore tested in pharmacological experiments.

A comparison of arthritis after experimental erysipelas, for example, in rats with rheumatoid arthritis in humans, reveals a broad overview of the individual morphological changes. The examinations were carried out according to the information given in Schulz et al., Contrib. Path. 15'i, 1-26, 27-51 (1975). The arthritis in rats caused by erysipelas can be reproduced with almost 100% certainty by a single injection. More than 6 joints are always altered in red gouty arthritis, large and small *

Limb joints in the same form. In experimental erysipelas, all animals are still highly poli after 3 months? erative processes.

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To test the substances from the manifestation phase, the following parameters are used to assess the effectiveness of the preparation:

Paws

The arthritis of the rat is clinically characterized in animals with a body weight of ljo g from the 3rd day at about 2oo g from the 3rd day by a high-grade peri-articular edema.

Kidney (egg shell)

Nephrosis caused by microthrombi can occur at approx.

3o - 4o% of the animals on 7 · - δ. Day occur.

eye

Inflammation (clouding) of the cornea around the 8th day.

External genital organs, tail tip Necrosis due to thrombi from 6th - 8th day.

aorta

Between the 6th and 11th day, fibrin-rich thrombi appear on the aortic intima in the rat. The largest area expansion is reached around the 8th day.

Male Wistar rats with weights between 15 o and 18 o g were used for the study. The animals were kept in individual cages and were given a rat standard diet (Ssniff R) and water ad libitum.

The room temperature was constant at 22 ° C, the relative air humidity between 5o and 6o%. The daily lighting duration was 12 hours.

Before the start of the test, the animals had an acclimatization period of 10 days.

Infection occurred with the T 28 erysipelas.

The dosage was 2 ml cutaneous (approx. Loo - 2oo million germs). The compounds to be tested here were in sterile physiological saline. r suspended and in a dosage of 5 mg / kg iP ..

applied. The treatment ran from Infecton day until the end of the experiment or the death of the animals, 5 times a week.

The evaluation of the characteristic aortic thrombi resulted in the following value

Control group S score 2.8 o

Sitostering glucoside: 2,000 hecogenin glucoside; 1.38

Diosgenin glucoside: 1.33.

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Claims (6)

V V * - 21 - 1 ") Medicament with effect as a prostagiandin synthetase inhibitor, characterized by a content of sterol glycosides and / or their esters and / or spiroketal steroid glycosides and / or their esters. 2.) Medicament according to claim 1, characterized by * a content of glucosides of T & Lölsterole, sitosterol, ergosterol, cholesterol, 5t ^ cholesterol, lanosterol, 24, 25 ~ dihydrolanosterols, stigmasterols and / or soyasterols. 3rd ) Medicament according to Claim 1, characterized by a content of β-D-galactoside, β-D-maltoside, β-D-lactoside and / or β-L> -cellobioside of sitosterol. 4 «) Medicament according to claim 1, characterized by a content of glucosides of diosgenin, hecogenin and / or tigogenin. 5 ·) Medicament according to claim 1-4, characterized in that the particle size of the active substance is in the range of less than 0.1 mm in diameter and in particular of about 0.60 mm and smaller diameter. + 6%