CH677732A5 - - Google Patents

Description

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description

The invention relates to a medicament preparation which acts on the cardiovascular system and which primarily acts on the thromboembolic conditions of the cardiovascular system. The invention also relates to a method for producing this pharmaceutical preparation.

It is known that the <n-3-unsaturated fatty acids with a chain length of 18-24 carbon atoms have advantageous biological properties. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are of particular importance. Dyerberg et al. Report on the importance of these two acids and their complex biological effects. (The Lancet, 15.117 / 1978 /).

Goodnight et al. Reported on the important role that the polyunsaturated fatty acids, primarily EPA and DHA, play in hyperlipidemic diseases and in thrombosis. a summary paper has been published (Arteriosclerosis 2,87 / 1982 / Review).

Pharmaceutical preparations containing the active substance EPA and DHA are - primarily to lower the cholesterol level in the blood - in DE-PS 3 438 630, furthermore as a means of preventing thrombi in cardiac patients and vein calcification in the brain in the published Japanese patent applications 58.08 037 and 60.49 097.

Numerous publications have appeared on the property of EPA and DHA to inhibit platelet aggregation and thus thrombus formation, for example by Spencer and Caraega (Prostagl. Leukotrienes and Med. 23, 129/1986 /) and by Knapp et al. (New Engl. Journ. Of Med. 314, 937/1986 /). •

The antiviral activity of EPA and DHA is described in U.S. Patent 4,513,008.

The active components of fish oil, including EPA and DHA, act as precursors in the PG-3 series biosynthesis, at the same time they inhibit the formation of the harmful metabolites, for example T XAg and T XBa, which are more complex in a chain arachidonic acid-based biochemical processes, the so-called «arachidonic acid cascade».

In addition to their numerous beneficial properties, the polyunsaturated fatty acids also have the disadvantageous pathological peculiarity of being decomposed in the human organism by spontaneous oxidation mechanisms, whereby active aldehydes, including malonaldehyde which is harmful to the organism, are formed. These aldehydes can primarily interact biologically with the connective tissues, which can lead to so-called “ceroid lipofuscinosis”.

In recent decades, research has started to deal with the biological effects of micro-elements and trace elements. It was recognized that selenium is one of the most important and essential substances for life. The beneficial effect of selenium is primarily due to the fact that it activates glutathione peroxidase, more precisely: its prosthetic group, which is the most important endogenous inhibitor of the harmful peroxidation processes.

The selenium is in itself an effective blood pressure lowering substance, it improves ischemic, hypoxic and infarct conditions of the heart and prevents the ceroid lipofuscinosis of the central nervous system. It is also effective against periodontitis (tooth root infection). Its effectiveness against cancer is significant, it has also been shown to prevent the possibility of cancer developing. Selenium can also be used as a mutagenic inhibitor.

Selenium is not accumulated in the organism, i.e. constant replenishment must be ensured. To date, the organism mainly absorbs selenium in the form of inorganic compounds such as SeOa and NagSeOa. Selenium deficiency causes liver necrosis, death of muscle tissue, destruction of the erythrocyte membrane, damage to the connective tissue, in the ECG S-T elevation as a change, and can also cause Kwashios syndrome and sclerosis multiplex.

A summary of the biological effects of selenium has been published, for example by Thressa et al. (Nutrition Review 35.7 / 1977 /), by Shamberger (J. of Env. Path. And Tox. 4, 305/1980 /) and by Masukawa et ai. (Experrenta 39,405 / 1983 /).

Organic selenium compounds can be prepared by preparative synthesis, as described, for example, by Klaiman and Günther in their book “Organic Sélénium Compounds, Their Chemistry and Bio-gy” (John Wiley and Sons, Inc. New York / 1973 /). Organic selenium compounds can also be produced with the help of microorganisms. The microorganisms are grown on selenium-enriched nutrient media. The suitable microorganisms absorb the selenium, incorporate it into their metabolism and bind it to organic substances, primarily to amino acids or fats (Danch and Chmielowsky: Pr. Nauk. Univ. Slask Katowich 1, 57/1985 / and Gennity et al .: Biochem. Biophys. Res. Commun. 118.176 / 1984 /).

Numerous microorganisms are known which are capable of collecting individual elements - including selenium - in their environment and accumulating in their organism. The preparation of yeasts enriched with selenium is described in US Pat. No. 4,530,846 and in published Japanese Patent Applications Nos. 60,224,451 and 57,174,098. The enrichment of other microorganisms, such as bacteria, radiation fungi and filamentous fungi, in addition to yeasts, with selenium is described in the published Japanese patent application No. 78,148,587.

The selenium-containing yeast is recommended in the published Japanese patent application No. 58.129 954 in a mixture with vegetable oils for the treatment of old-age diseases.

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The aim of the invention was to develop a pharmaceutical preparation which combines the advantageous effects of DHA and EPA with those of selenium-containing organic natural compounds produced by means of special microorganisms and at the same time is free from the disadvantageous properties of the polyunsaturated fatty acids.

The invention is based on the knowledge that this goal can be achieved and a means suitable for eliminating the ceroid lipofuscinous processes can be produced by combining selenium-containing, non-toxic human microorganisms with polyunsaturated fatty acids.

The invention accordingly relates to a medicament preparation which acts on the cardiovascular system and which contains 0.5-50% by mass of selenium-containing microorganisms and 99.5-50% by mass of at least double-unsaturated fatty acids with a chain length of 18-24 carbon atoms and / or their derivatives , if desired in combination with the usual extenders and / or carriers and possibly antioxidants.

The invention further relates to a method for producing the pharmaceutical preparation described. It is characteristic of the process that 0.5-50% by mass of selenium-containing microorganisms, 99.5-50% by mass of at least double-unsaturated fatty acids with a chain length of 18-24 carbon atoms and / or their derivatives and, if desired, those customary in pharmaceutical production Extenders and / or carriers and optionally antioxidants mixed together and formulated into medicinal products.

The pharmaceutical preparations according to the invention preferably contain eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and selenium-containing yeast.

As the base material for the cù-3-unsaturated fatty acids with a chain length of 18-24 carbon atoms, which preferably form a component of the preparation, they come from freshwater and saltwater fish, primarily from mackerel, cod, herring, sardines, octopus and their liver Fish-extractable oils in question, for example cod liver! (Cod liver oil) and shark liver oil.

In addition to EPA and DHA, the fish oils contain large amounts of saturated fatty acids and unsaturated fatty acids as well as unsaponifiable components. These components must be removed because otherwise the preparation according to the invention, used in higher doses, would increase the calorie intake and the triglyceride level in the blood. In addition, the non-saponifiable components can contain steroids or cholesterol, vitamin D (its provitamin) and vitamin A. Vitamins A and D are accumulated in the human body; With a preparation containing these vitamins, long-term treatment necessary for the desired effect could not be carried out. It is therefore advisable to remove the components mentioned from the fish oils (saturated fatty acids, monounsaturated fatty acids, non-saponifiable components such as cholesterol, vitamin A, vitamin D). In this way, EPA and DHA are enriched in fish oil to (together) more than 50%.

As microorganisms, strains that are not toxic to humans are preferably used. The genera which are also widely used in biotechnology are most suitable for this, such as Lactobacillus, Leuconostoc, Pediococcus, Acetobacter, Streptococcus, Torula, Kluyverromyces, Candida, Brettanomyces, Brevibacterium, Saccharomyces, Torulopis, Pichia, Hansenula, Oidium, Rhidium Trichospora, Pénicillium, Rhizopus, Mucor, Monascus, Aspergillus and other species suitable for human consumption,

The preparation according to the invention can contain, as an anti-oxidant active preservative, a-Tokoferol (vitamin E), glutathione or conventional antioxidants, for example butylated hydroxytoluene.

The additives, for example lactose, starch or magnesium stearate, which are customary in pharmaceutical production are used as carriers, extenders and other auxiliaries.

The pharmacological studies showed that the preparation has no harmful side effects. Even a dose that was 100 times the amount found in the microsomal enzyme system of the rat did not cause any changes. Treatment with the preparation significantly reduced the amount of lipofuscins accumulated in the organism compared to the untreated control. The inhibition of platelet aggregation was clearly demonstrated in female Wistar rats in tests lasting 6 weeks.

The optimal daily dose of the preparation is 2 g based on an average body weight of 70 kg. The oil component contains an average of 22% EPA and 43% DHA. The optimal daily selenium dose is 100-300 [ig.

The advantages of the preparation according to the invention can be summarized as follows:

1. It combines the beneficial properties of EPA and DHA with those of selenium and yeast.

2. It reduces or eliminates the harmful effects of the known fish oil-containing preparations, which result from the content of saturated lipoid components, for example sterols, and vitamins A and D.

3. When taking the preparation, you do not have to expect ceroid lipofuscinosis, as occurs through the consumption of polyunsaturated fatty acids.

4. The selenium is enriched as a natural substance in microorganisms, for example in bacteria,

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Mushrooms, yeast included. The selenium ingested together with the special microorganism is better absorbed and exerts its beneficial effects better.

5. The organic selenium compounds can be enriched in microorganisms, primarily in yeast, on an industrial scale and cheaply.

6. The preparation is effective against atopic disorders and can also be used for preventive treatment against eczema, asthma, allergic symptoms, allergic rhinitis and against disorders with atopic connections, for example migraines, Crohn's syndrome, ulcerative colitis, otitis media, nephrotic syndromes, Diabetes can be used. In particular, it is effective against disorders of the circulatory system, apoplexy, thromboembolic conditions such as cerebral hemorrhage, infarction, Keshan syndrome in adolescents, and is also suitable for preventing the development of pathological conditions.

The (o-3-unsaturated fatty acids used as a component of the preparation according to the invention can be prepared in the manner described in Example 1, and the selenium-enriched microorganisms according to Examples 2-6. Examples 7-9 show the preparation of the preparation itself.

example 1

24 kg of mackerel oil are dissolved in 16 liters of methanol at 50-60'C. 8 kg of 40% sodium hydroxide solution are added to the solution with stirring, and the mixture is stirred at 60 ° C. for a further 45 minutes. At about 60 ° G, 20 kg of 15% hydrochloric acid are added to the solution. The phases are separated and the organic phase is washed neutral with 10 kg of 15% hydrochloric acid and then with 180 liters of hot tap water. The resulting phases are separated again. The oily phase is mixed with 100 liters of acetone, then heated to 45 ° C., the solution of 3.8 kg of LÌ0H-H2C! N 30 liters of water is added and the mixture is left to stand overnight after half an hour of stirring. The next day is filtered, the acetone filtrate is evaporated. The residue is acidified with 8 kg of 15% acidic acid, extracted three times with hexane and the hexane phase is evaporated. The entire cleaning process is carried out under a nitrogen atmosphere. At the end you get 6.4 kg of purified fish oil.

One kg of the mackerel oil purified in the manner described is added dropwise to the solution of 3 kg of urea in 9 liters of methanol at 60 ° C. The mixture is stirred at the same temperature for 2 hours. After cooling, the mixture is left in the freezer at -10 ° C. overnight. The next day is filtered and the filtrate evaporated. 2.5 liters of semi-dilute hydrochloric acid are poured into the evaporation residue, then it is shaken out with hexane and the hexane phase is washed neutral with water. After drying over sodium sulfate, the solution is evaporated. In this way, 0.34 kg of w-3-unsaturated fatty acids are obtained. Iodine number: 315; EPA content: 24%, DHA content: 42%.

Example 2

Glass balloons with a volume of 5 liters are each filled with 2 liters of sterilized, liquid nutrient medium containing malt extract and enriched with 3% by mass of glucose. After adding 5 μg / ml sodium selenite, the batch is inoculated with Saccharomyces cerevisiae and then cultivated with aeration at 32 ° C. for 72 hours. In the 48th hour a further 5 ng / ml sodium selenite are added to the culture. After filtering, the cells are washed several times with distilled water and then dried at 68-70 ° C. The material is micronized and the selenium content is determined using the atomic absorption method. Se content: 2600 ng / g.

Example 3

Erlenmeyer flasks with a volume of 500 ml are filled under sterile conditions with 100 ml of liquid nutrient medium containing yeast extract. After adding 5 ng / ml sodium selenite, the mixture is inoculated with the Aspergillus sojae mushroom and then incubated at 28-30 ° C on the shaking table. On day 3, a further 5 ng / ml sodium selenite is added to the culture. After 5 days, the solid substance is filtered off, washed and dried in the manner described in Example 2. The Se content determined as in Example 2 is 3000 ng / g.

Example 4

The procedure is as described in Example 3, but Torulopsis utilis yeast is used. The selenium content of the microbe mass is 1299 ng / g.

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Example 5

Streptococcus thermophilus is cultivated at 45 ° C. in the nutrient medium prepared according to Example 3. The microbial powder obtained after working up has a selenium content of 4200 μg / g.

Example 6

Lactobacillus mesenteroides is cultivated in the manner described in Example 3. The nutrient medium is supplemented by 1% glucose and 2% calcium carbonate. Selenium content of the bacterial mass: 3000 (xg / g.

Example 7

10 g of selenium-containing yeast powder (Se concentration: 2600 ng / g) are added to 150 g of enriched mackerel oil (EPA content: 24%, DHA content: 42%). The mixture is homogenized and preserved by adding 0.15 g of vitamin E. The active ingredient mixture is filled into pearl capsules made of soft gelatin or into soft gelatin capsules and packed in blister film.

Example 8

The procedure is as described in Example 7, but the following starting materials are used: 400 g of 65% enriched cod liver oil (EPA content: 22%, DHA content: 43%), 100 g yeast powder with a Se content of 1200 ng / g and 0.4 g vitamin E. The homogenate is filled into capsules suitable for taking 500 mg of active ingredient.

Example 9

The procedure described in Example 8 is followed, with the difference that the fish oil component is 400 g of 30% fish oil (EPA content: 18%, DHA content: 12%; manufacturer: Martens and Co., Bergen, Norway) used.

Example 10

The procedure is as described in Example 8, with the difference that 400 g of fish oil which contains 9.2% EPA and 10.4% DHA are used as the fish oil component.

Example 11

Known devices and processes are used to produce tablets of the following composition:

Enriched cod liver oil 200.0 mg (DHA content: 43%, EPA content: 22%, vitamin E content: 0.1%)

Se yeast powder (Se content: 1500 p.g / g) 86.0 mg

Lactose 140.0 mg

Strength 60.0 mg

Polyvlnyipyrrolidone 3.5 mg

Magnesium stearate 3.5 mg

If desired, the tablets can be coated with sugar or other substances on a coating machine.

Claims (8)

Claims 1. Cardiovascular drug preparation, characterized in that it contains 0.5 to 50% by mass of selenium-containing microorganisms and 99.5 to 50% by mass of at least double-unsaturated fatty acids with a chain length of 18 to 24 carbon atoms. 2. Medicament preparation according to claim 1, characterized in that it contains 5 to 35% by mass, preferably 15 to 25% by mass, of selenium-containing microorganisms. 3. Medicament preparation according to claim 1 or claim 2, characterized in that it is 95 to 5 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 677 732 A5 Contains 65% by mass, preferably 85 to 75% by mass, of at least double unsaturated fatty acids with a chain length of 18 to 24 carbon atoms. 4. Medicament preparation according to one of the preceding claims, characterized in that it contains unsaturated fatty acids from the oil of sea fish. 5. Medicament preparation according to one of the preceding claims, characterized in that it contains 5,8,11,14,17-eicosapentaenoic acid and 4,7,10,13,16,19-docosahexaenoic acid as unsaturated fatty acids. 6. A process for the preparation of a medicament preparation which acts on the cardiovascular system, characterized in that 0.5 to 50% by mass of selenium-containing microorganisms and 99.5 to 50% by mass of at least two times unsaturated fatty acids with a chain length of 18 to 24 carbon atoms formulated into medicinal products. 7. The method according to claim 6, characterized in that one uses unsaturated fatty acids from the oil of sea fish. 8. The method according to claim 6 or 7, characterized in that 5,8,11,14,17-eicosapentaenoic acid and 4,7,10,13,16,19-docosahexaenoic acid are used as unsaturated fatty acids. 6