NL8800693A - PHARMACEUTICAL PREPARATIONS WITH AN EFFECT ON THE HEART AND ON THE CARDIOVASCULAR SYSTEM AND THE METHOD OF PREPARING THE SAME. - Google Patents

Description

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Pharmaceutical preparations having an effect on the heart and on the cardiovascular system, and a method for their preparation.

The present invention relates to pharmaceutical compositions having an effect on the heart and the cardiovascular system.

According to another aspect of the invention, it concerns a process for the preparation of these preparations.

It is known that unsaturated fatty acids have favorable biological properties. Of these compounds, eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) are excellent. Dyerberg et al / The Lancet jj>, 117 (1978} γ) have pointed out the importance and multiple biological action of both acids.

The action, associated with the important role of polyunsaturated fatty acids, especially EPA and DHA, in the hyperlipidaemia and thrombotic disorders, has been summarized by Goodnight et al. / Arteriosclerosis 2, 87 (1982) Review / ·

Pharmaceutical compositions containing EPA and DHA as active ingredients are described, for example, in German Patent No. 3,438,630 for lowering blood cholesterol levels, further in published Japanese Patent Applications Nos. 58,08037 and 60,49097 against cerebral sclerosis and for prevention of thrombosis formation in patients suffering from heart disease.

Various publications are devoted to the clotting of platelet-inhibiting and thus thrombotic-inhibiting activity of EPA and DHA / Spencer and Caraega: Prostagl. Leucotrienes and Med. 23, 129 (1986); Knopp et al .: New Engl. Journ. or Med. 314, 937 (1986)].

The antiviral activity of EPA and DHA is described in U.S. Patent No. 4,513,008.

The active ingredients of the fish oil, eg.

EPA and DHA are precursors for the biosynthesis of the PG-3 series and prevent the formation of harmful metabolites, such as TXA2 and TXB2, resulting from the so-called "arachidonic acid step 35", which is a chain of complex biochemical processes. 880 009 3 - 2 - * 1 sen, starting from arachidonic acid.

In addition to their various beneficial effects, the polyunsaturated fatty acids have the disadvantageous pathological property of being subject to spontaneous oxidative decomposition in the human organism and giving rise to the formation of active aldehydes, such as malondialdehyde, which are harmful to the organism. These aldehydes are capable of reacting mainly with connective tissues in a physiologically harmful manner, which may lead to the so-called "ceroidal lipofuscinosis".

It is further known that algae have been used for a long time by humans for food and feed purposes. Algae are therefore mainly consumed by peoples from the Far East; however recently they have been widely used in dry form or in tablet form also in developed countries.

Algae are the carriers of very valuable nutrients, since their dry form contains a high concentration of substances essential for healthy living, such as vitamins, proteins, complexes of proteins with microelements, saccharides, polyunsaturated fatty acids and the like .

General and detailed information regarding algae has been described, e.g., by Zajic in: Properties and Products of Algae (Edition Planum, New York, 1970).

Research into the biological action of microelements and trace elements has been started in the last decades. It has become known, therefore, that one of the most important and indispensable substances of living mechanism is. The beneficial effect of selenium is mainly based on its activating action on the glutathione peroxidase enzyme, since selenium is an essential component of the prosthetic group of the glutathione peroxidase enzyme. This enzyme is the most important endo-35 gene inhibitor of the harmful peroxidation processes.

Selenium as such has a hypotensive effect, improves the. ischemic, hypoxic and infarct states of the heart and prevents the cerebral lipofuscinosis of the central nervous system; in addition, selenium exercises a gun .8800693 - 3 -

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► strong effect on periodontitis. Selenium also has a significant anti-cancer effect and has been shown to reduce the likelihood of the development of cancer diseases; moreover, it is considered to be a mutagenesis inhibitor.

5 Selenium is not accumulated in the organism, which means that it must be continuously supplemented. To this end, selenium was mainly introduced into the organism in the form of inorganic compounds, e.g., selenium dioxide and sodium selenite. A number of abnormalities or 10 diseases, such as liver necrosis, myonecrosis, erythrocyte membrane destruction, interstitial lesions, ST elevation in the EGC, Mulberry's heart syndrome, kwashiorcor syndrome (protein malnutrition) and multiple sclerosis have been induced by a lack of selenium.

An extensive report on the biological action of selenium was published by Thressa et al.

/ Nutrition Review 35, 7 (1977) _ /, Shamberger [j. or Env. Path, and Tox. £ 305 (1980) J as well as Masukawa et al. / Experientia 39,405 (1983) J.

The object of the present invention is to provide a new pharmaceutical composition, which mainly acts on the heart and the cardiovascular system, enabling the combination of the beneficial properties of DHA, EPA, algae as well as of selenium and at the same time eliminating the adverse properties of polyunsaturated fatty acids.

The invention is based on the recognition that the above object can be perfectly accomplished by using a selenium containing algae together with polyunsaturated fatty acids.

Accordingly, the invention relates to a pharmaceutical composition having an effect on the heart and the cardiovascular system, which composition contains 0.5 to 50% by weight of selenium-containing algae in addition to 99.5 to 50% by weight c <3. .22 fatty acids with at least two unsaturated bonds or their derivatives, optionally mixed with carriers and / or additives and / or antioxidants, which are commonly used in the pharmaceutical industry.

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In addition, the invention relates to a process for the preparation of the above preparation, characterized in that 0.5 to 50% by weight of selenium-containing algae, as well as 99.5 to 50% by weight of C18_22 fatty acids with at least two unsaturated its compounds or derivatives, optionally together with carriers and / or additives and antioxidants commonly used in the pharmaceutical industry, are mixed together.

The compositions according to the invention advantageously contain eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) as well as a selenium-containing alga.

The selenium-containing alga is prepared in the manner described in the related patent application Serial No. ........ (Hungarian patent application No. 575/88).

15 As starting material for the c-22-unsaturated fatty acids, which form one constituent of the preparation, oils from various sea fish and freshwater fish, mainly from mackerel, cod, herring, sardine, squid and liver, these fish, e.g. liver-20 tear and shark liver oil.

In addition to EPA and DHA, the fish oils contain a large amount of saturated fatty acids as well as fatty acids, which are unsaturated to a low level, and other non-hydrolysable components. The removal of these components is very important, since the dosage of the therapeutic compositions prepared from the fish oils sensitively increases the introduced amount of calorie as well as the blood triglyceride level. In addition, the non-hydrolyzable components may contain steroids, such as cholesterol, vitamin D (and its provitamin) and / or vitamin A. Vitamins D and A are accumulated in the human organism, whereby an extended treatment required for the desired effect would be impossible using a formulation containing these vitamins. As a result, the above-mentioned ingredients, such as the saturated and partially unsaturated fatty acids, as well as the non-hydrolyzable ingredients, e.g., cholesterol, vitamins A and D, are removed from the fish oil. In this way, the total amount of EPA and DHA in the fish oil is enriched to more than 50%.

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Chlorella or Scenedesmus strains can advantageously be used as algae, which are suitable for human consumption and, on account of their very favorable biological action, form an indispensable starting material for the modern diet.

To inhibit the oxidation of the preparation according to the invention, use is made advantageously of α-tocopherol (vitamin E), glutathione or the traditional antioxidants, such as butylhydroxytoluene, as active preservatives.

As a liquid or solid carrier, the materials commonly used in the pharmaceutical industry, such as lactose, starch or magnesium stearate, can be used.

Pharmacological studies have shown that the composition is free of side effects that would be harmful to health since no change was observed in the rat liver microsomal enzyme system using a hundredfold of the usual dose. The amount of lipofuscin stored in the organism was significantly reduced compared to the control test after treatment with the composition of the invention. Based on studies performed using the preparation according to the invention in 25 female Wistar rats for 6 weeks, an unambiguous inhibitory effect on platelet clotting was observed.

The optimal effective daily dose (calculated for an average body weight of 75 kg) of the preparation, consisting of fish oil and algae powder, is 30 to 2 g with a selenium content of 240 jig. On average, the oil component consists of 22% EPA and 43% DHA.

The main advantages of the preparation according to the invention can be summarized as follows: a) The preferred properties of EPA and DHA as well as of selenium and the alga are combined.

b) The harmful effects caused by the saturated lipid components of the known fish oil-containing preparations as well as the vitamin A and D content have been eliminated.

.8800693 - 6 -ψ (c) The possibility of "ceroidal lipofuscinosis", which occurs with the consumption of polyunsaturated fatty acids, has been removed.

d) The preparation contains selenium as a natural substance, enriched in the alga; the selenium, administered in conjunction with the alga, is better absorbed and more advantageously exerts its beneficial effect.

e) The preparation develops a suitable therapeutic effect in case of atopic disturbances and is suitable for the preventive treatment of eczema, asthma, allergic symptoms, allergic rhinitis and / or atopic abnormalities, eg migraine, Crohn's disease, ulcerative colitis, otitis media, nephrotic syndrome and diabetes.

The composition according to the invention is particularly suitable for the treatment of abnormalities of the cardiovascular system; for apoplectic conditions, thromboembolic conditions, such as stroke, infarction and Keshan's syndrome in young patients, as well as for the prevention of abnormal conditions.

The invention is further illustrated by the following non-limiting Examples.

The ω-3 polyunsaturated fatty acids used in the composition of the invention can be prepared according to Examples I and II, while the selenium-enriched algae can be prepared according to Example III. The pharmaceutical preparations according to the invention are described in Examples IV-VI.

EXAMPLE I

2 kg of sodium hydroxide were dissolved in 70 l of 95% ethanol, after which 10 kg of cod liver oil was added at a temperature of 50 to 60 ° C. The mixture was refluxed under nitrogen for 2 hours and then cooled to 10 ° C with stirring. The sodium salt of the saturated fatty acids was precipitated. The crystals were filtered and washed with a little ethanol. The ethanolic filtrate was evaporated and 20 L of boiled water was added to the residue. The non-hydrolyzable compounds, such as cholesterol, have been completely removed by extraction with 5 L of hexane.

The aqueous phase was acidified to a pH of 2 by adding sulfuric acid (8800693-7) and extracted again with 15 L hexane. The organic phase was washed with water. dried over anhydrous sodium sulfate and then evaporated to yield 3.2 kg of a concentrated oil with a DHA content of 36.8% and an EPA content of 31.8%. This oil was brown and smelled like fish. The oil was then mixed with Puller's soil, heated under nitrogen at 105 ° C for 10 min and then filtered hot. Deodorization was accomplished via steam distillation under vacuum at 170 C / l Hgmm for 10 hours to yield 1.6 kg of a pale yellow, tasteless, odorless oil of unaltered composition.

EXAMPLE II

8 kg of 40% sodium hydroxide solution was added dropwise at 50 to 60 ° C to 24 kg cod liver oil, dissolved at 60 ° C in 16 1 methanol with stirring. Then the mixture was stirred at 60 ° C for an additional 45 min. 20 kg of 15% hydrochloric acid was added to the solution at about 60 ° C.

After separation, the organic phase was washed with 10 kg of 15% hydrochloric acid and then with 180 L of hot water until the solution became neutral. The phases were again separated, after which 100 l of acetone was added to the oil phase, which mixture was then heated to about 45 ° C, followed by the addition of a solution of 3.8 kg of lithium hydroxide monohydrate in 30 l of water. After stirring for 30 min, the mixture was allowed to stand overnight, followed by filtration, and the acetone filtrate was evaporated. The residue was acidified by addition of about 8 kg of 15% hydrochloric acid and then extracted with hexane 3 times and finally evaporated.

A nitrogen atmosphere was maintained throughout the purification operation. 6.4 kg of purified fish oil with an iodine value of 258 and an acid number of 160 were obtained.

One kg of the cod liver oil, purified as described above, was added dropwise at 60 ° C to a solution of 3 kg of urea in 9 L of methanol. The mixture was stirred at the same temperature for 2 hours and then cooled. Then the mixture was stored at -10 ° C overnight, then filtered and the filtrate evaporated.

2.5 11: 1 hydrochloric acid was added to the residue, after which the mixture was stirred for an additional 15 min. After extraction with hexane .8800693 i - 8 - *, the hexane phase was washed with water until neutral, then dried over anhydrous sodium sulfate and finally evaporated to give 0.34 kg of 3-3 unsaturated fatty acid with an iodine value of 315 which is 24% EPA and 42% DHA.

EXAMPLE III

40 mg of sodium selenite was added to 8 L of Bud-Pringsheim's culture medium, which was transferred to a 10 L algae culture glass bottle. The culture medium thus obtained was sterilized at 121 ° C under an overpressure of 1 bar for 10 min. Then the sterile solution was cooled and inoculated with a pure culture of Scenedesmus obtisiusculus, which is able to absorb selenium well. Sterile air containing 5% by volume of carbon dioxide was bubbled through the culture medium at 25 ° C, while the system was exposed using an electric discharge tube operating at 4000 lux at a wavelength of 440-700 µm. After a culture period of 14 days, the alga was separated from the culture medium.

The algae mass thus obtained was decomposed by sound waves and then carefully dried at a temperature below 65 ° C. The yield was 6 g of an algae powder with a selenium content of 1200 µg / g.

EXAMPLE IV

100 g of a selenium-containing algae powder (selenium content: 260 µg / g) was added to 150 g of 65% enriched cod liver oil (containing 22% EPA and rel DHA). After homogenization, 0.1% vitamin E was added. The active ingredient thus obtained was transferred into soft gelatin capsules and then packed in blister foil.

EXAMPLE V

The procedure described in Example IV was repeated, except that the following starting materials were used: 400 g of enriched cod liver oil with an EPA content of 22% and a DHA content of 43%; 70.6 g of an algae powder with a selenium content of 1200 yag / g; and 0.4 g of vitamin E. The homogenizate was transferred to capsules capable of absorbing up to 500 mg of the active ingredient.

EXAMPLE VI

Tablets of the following formulation were prepared using known pharmaceutical devices. 8800693 4-9 and methods:

Cod liver oil enriched with EP A and DHA and containing 0.1% vitamin E (with a DHA content of 43% and an EPA-5 content of 22%) 200 mg

Selenium-containing algae powder (selenium content: 1500 yag / g) 86 mg lactose 140 mg starch 6 0 mg 10 polyvinylpyrrolidone 3.5 mg magnesium stearate 3.5 mg

If desired, the tablets can be coated with sugar in a tableting machine.

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Claims (10)

Pharmaceutical preparation with an effect on the heart and the cardiovascular system, characterized in that this preparation contains 0.5 to 50% by weight of selenium-containing algae in addition to 99.5 to 50% by weight of C.jg_22 fatty acids with at least two unsaturated bonds or their derivatives, optionally mixed with carriers and / or additives and antioxidants, which are commonly used in the pharmaceutical industry. 2. A composition according to claim 1, characterized in that it contains 5 to 35% by weight of selenium-containing algae. 3. A composition according to claim 1, characterized in that it contains 15 to 25% by weight of selenium-containing algae. 4. A composition according to claim 1, characterized in that it contains 95 to 65% by weight of c-8-22 fatty acids with at least two unsaturated bonds or derivatives thereof. 5. Preparation according to claim 1, characterized in that it is 85 to 75% by weight c- 8-22 fatty acids with at least two unsaturated bonds or derivatives thereof. 6. Pharmaceutical preparation according to any one of the preceding claims 1-5, characterized in that this preparation contains as unsaturated fatty acids fatty acids extracted from the sea fish oil. Pharmaceutical preparation according to claims 1-6, characterized in that as unsaturated fatty acids in this preparation contains 5,8,11,14,17-eicosapentanoic acid and 4,7,10,13,16,19-docosahexanoic acid. Method for the preparation of a pharmaceutical composition having an effect on the heart and the cardiovascular system, characterized in that 0.5 to 50% by weight of selenium-containing algae and 99.5 to 50% by weight of 35 ci8- 22 fatty acids with at least two unsaturated bonds or derivatives thereof, optionally with carriers and / or additives and .8800693 * - 11 - antioxidants commonly used in the pharmaceutical industry. 9. A method according to claim 8, characterized in that 5 to 35% by weight of selenium-containing algae and 5 95 to 65% by weight of C-22 fatty acids with at least two unsaturated bonds are mixed together. 10. Process according to claim 8, characterized in that 15 to 25% by weight of selenium-containing algae and 85 to 75% by weight of C18-22 fatty acids with at least two unsaturated bonds are mixed together. . 8 8 0 0 6 S 3