GB2203042A - Compositions acting on the heart and cardio-vascular system - Google Patents

Description

2 2 0 3 C 12 PHARMACEUTICAL COMPOSITIONS ACTING ON THE HEART AM

CARDIOVASCULAR SYSTEM AND PROCESS FOR PREPARING SAME This invention relates to pharmaceutical compositions acting on the heart and cardiovascular system.

According to an other aspect of the invention, there is provided a process for the preparation of these compositions.

It is known that C 18-22 Ccr-3 unsaturated fatty acids possess advantageous biological properties. Of these compounds eicosapentaenoic acid (EPA) and docesahexaenoic acid THA) are outstanding. Dyerberg et al. /The Lancet 15, 117 (197ST pointed at the importance and multifold biologizal effects of both acids.

The effects c=nected with the important role of polyunsaturated fatty acics, particularly of EPA and CHA, in the hyperlipideemia and thrcmbotic diseases have been summarized,' by Goodnight et al. /-Arteriosclerosis 2, 87 (1962) ReviehE Pharmaceutical compositions containing EPA and DHA as active ingredients have been described e.g. in the German patent specification No. 3,43S,630 for lowering the blood cholesterol level, further in the published japanese patent applications Nos. 58.08037 and 60.49097 against cerebral sclerosis and for preventing the thrombus formation in patients suffering from heart diseases.

Several papers have been devoted to the platelet-aggregation inhibiting and thus the t%rombus-formation inhinit- 1 ing effect of EPA and DHA /-SQencer and Caraega: Prcstagl. Leucctrienes and Med. 23, 129 (1986); Knopp et al.: New Engl. Journ of Med. 314, 937 \110,86)7.

The antiviral action of EPA and DHA is described in the United States patent specification No. 4,513,006.

The active ingredients of the fish-oil, e.g. EPA and DHA, are precursors of the biosynthesis of the PG-3 series and they inhibit the formation of harmful metabolites such as TXA 2 and TXB 2 arising from the so-called "arachidonic acid cascade" which is a chain of complicated biochemical processes starting from arachidonic acid.

In addition to their several favourable effects, the polyunsaturated fatty acids have the disadvantageous pathologic property that they are subjected to a spontaneous oxidative decompcsition in the hLman organism and thus they give rise to the formation of active aldehydes such as malondialdehyde which is harmful to the organism. These aldehydes are capable to react mainly with the connective tissues in a physiologically harmful way which may lead to the so-called "ceroidal lipofL,scinosis".

It is further known that algae have from time immemorial been used by the mankind for the purpose of nutrition and feeding. Thus, algae are mainly consumed by the peoples of the Far East; recently, hcwever, they are widely utilized in dry form or in the form of tablets in the developed countries, 4.

Algae are t-he carriers of highly valuable nutIritive r,atEr-ia'..s since their dried form contains high conc-entraticris of substances which are essential for healthy life such as vitamins, proteins, co-miplexes of prcteins with micrcelements, saccha:'Ldes, polyunsaturated fatty acids and the like.

General and detailed informations concerning algae are described e.g. by Zajic -in: Properties and Products of Algae (Edition Planum, New York, 1970).

Investigations on the biological effects of micro- elements and trace elements have been started in the 'Last decade. Thus, it became known that selenium is one of the most import-ant and.4ndispensa'L-:',e substances of life. The beneficial action Gf selenium is mainly ',-.,ased on its activat ing effect directed to the 91u4L-ath'Lone-pe-.oxidese enzyme as sel-enium is an JLnd..-.,pensable constituent of the prosthetic group -If the gluta"LI,-i-ione-peroxidase enzyme. This enzyme is the r-,ost endogenic inhititor of the harmful peroxidation processes.

Selenium in itself has hypotensive effect, improves the ischaemic, hypoxic and infarction states of the heart and inhibits the cercidal lipofuscinosis of the central nervous system; furthermore, it alsb.exerts a beneficial effect on periodontitis. It has a significant anticancer activity and proved to diminish the probability of the development of cancer diseases; fithermore, it is considered to be a mutapenesisinhibiting agent.

Selenium is not accumulated in the organism, thus it should continuously be supplEmented. Hillherto, selerium has rriainly been introduced to the croan,Lsr-. in the form of inorganic compounds, e.g. selenium dioxide and sodium selenite. A number 0 alte:ations or diseases, reseectively, such as liver necrosis, myonecrosis, oestruction of the erythrocyte membrane, interstitial laesions, ST-elevation in the ECG, Mulberry's heart syndrome, kwpshiorkor syndrome (protein malnutrition) and multiplex sclerosis proved to be induced by selenium deficiency.

A comprehensive report on the biological effects of selenium was published by Thressa et al. /-Nutrition Review 35, 7 (1977% Shamberger TY of Env. Path. and Tox. t, 305 (19017 as well as Masukawa et al. TExperientia 39, 405 (19ST7 The aim of the p:esent invention is to provide a novel pharmaceutical c=position, mainly acting on the heart and cardievescula: system, which renders possible the combination 0 the advantageous properties of DHA, EPA, algae as well as selenium anj which simultaneously eliminates the disadvantagenus p=perties of polyunsaturated fatty acids.

The invention is based on the recognition that the above aim can be perfectly achieved by using a selenium-containing alga together with polyunsaturated fatty acids.

Thus, the present invention relates to a pharmaceutical composition acting on the heart and cardiovascular system, which comprises 0.5 to 50% by mass of selenium-containing alga as well as 99.5 to 50% by mass of C 18-22 fatty acids containing at least two unsaturated bonds or the derivatives thereof, cpti2Pally in admixture with carriers and/or additives and/or anticxidants commonly used in the pharmaceutical industry.

Furthermore the invention relates to a process Cr preparing the above composition, which comprises mixing 0. 5 to 50% by mass of se Lenium-containing alga as well as 99.5 to 50% by mass of C 18- 22 fatty acids containing at least two unsaturated bonds or the derivatives thereof optionally together with carriers and/or additives and antioxidants commonly used in the pharmaceutical industry.

The compositions according to the invention suitably comprise eicosapentaenic acid (EPA) and docosahexaenic acid (DHA) as well as a selenium-containing alga.

The algae car., be prepared by cultivating -7 algae on or in an aqueous medium 'D to 2 x 10 moles per i--e cf a sclube organ.c a:-,di'or inorganc co-.".%und; preferatly the algae used fGr cultivat-Lon Is selected by treating a strain of algae N-.-ethy' -NI dine and propagatIng the strain on a se-e-.-..i---ccnta-J--J'ng propagation med-Lu-r, and selecting for further those '-'nd-v.,l.,ia'Ls that both incorporate selenium read2-,lv and have a high grzwth rate. Preferred algae species include unicellular blue or green algae, e.g. Chlorella sp., Scenedesr.us sp., sp., and in particular Aphanocapsa Chlorella C'--,crella fusca, Scenedesm.us ol.--t-is-'.iscu.-,us, Scenedesmus ob-ijcuu-c or Nostoc cG:-mune (as desc.ribed in our co-pend-ing anpl-- 'cation correspondling ",j dU In this way, se.Len--.,,ir-, acc.jr---,lates in the crg-anc-r. of the g-ow--'rg algae.

As raw material of the Cl,-22 ci-r-3 unsaturated fatty acids representing one compo-nent of the composition, oils obtainable from various marine and fresh-water fishes, mainly from mackerel, cod-fish, herring, sardine, squid as well as from the liver of these fishes, e.g. cod-liver oil and shark-1i,,.er oil, can be used.

1 k> In addition to EPA and DHA, the.fish-oils contain a large amount of saturated fatty acids as well as fatty acids unsaturated to a low extent and other nonhydrolyzable components. The removal of these constituents is very important since the dosis of the therapeutical compositions prepared from the fish-oils would sensitively enhance the introduced calory quantity as well as the blood triglyceride level. Besides, the nonhydrolizable constituents may contain such as cholesterol-, vitamin 0 (and its prcvitamJLn) vitamin A. The vitamins D and A are accumulated in -7 the human organism whereby a pro'longed treatment, needed tz, L the desire,-' effect, would be -'Lnpcss.ib'ie by using a ccr-..posijL., i:)n containing these vitarins. There"fore, the components mentloned above such as the saturateCJ ard partly unsaturated fatty acids as well as the nonhydrolyzable constituents, e.g. cholesterol, vitamins A and 0, are removed from the fish-oil. In this way the total amount of EPA and DHA in the is enriched to more than 50%.

As algae, Chicrella or Scenedesmus strains may suitably be used which are useful fcr the human ccn-cumption and, owing to their very favourable biological effect, represent an indispensable raw material of the modern alimentat-icn.

-he oxidation o' the -ompositon accord- FoT- in'-jib-'t-in2 4. 1 ing to the invention, it is suitable to use OC-tccoferol (vitamin E), glutathione or the traditional antic).xidants such as 'LJutylhydroxytoluerie as active preservating agents.

As vehicle or carrier, the materials commonly used in the pharmaceutical, -Lndustry such as lactose, starch or magnesium stearate can be used.

Pharmacological investigations proved that the composition is free from side effects.damaging the health, as no change was observed in the microsomal enzyme system of the rat liver by using a hundred-fold quan4k. ity of the usual dose.

The amount of liPofuscine accumulated in the organism was Significantly decreased as compared to the control after trea""ment with the composition. Based on the invest.-1c_a'tions carried out by using the composition of the invention on Alemale Wistar rats for 6 weeks, an platelet- 9 -aggregation inhibiting effect was observe14 The optimum effective daily dcse (calculated for an average body-weight of 75 kg) of the composition consisting of fish-oil and alga powder amounts -to 2 9, with a selenium content of 240 /ug. In average the oil component consists of 22% of EPA and 43% of DHA.

The main advantages of the composition according to the invention can be summarized as follows: a) The preferable properties of EPA and DHA as well as of the selenium and the alga are combined. b) The harm.'L'ul effects caused by the saturated lipid components of the known fish-oil containing compOsitions as well. as the v.+lar.in A and 0 content are eliminated.

Pcssibility of "ceroidel lipc.fuscincsis" occurring on consL:mption of polyunsaturated.fatty acids is abolished.

compositien contains selenium as natural substance enriched in the alga; the selenium administered together wit, the alga is better resor-bed and exerts more preferably its favourable effects. e) The composition develops a fav6urable therapeutic action in case of atopic disturbances and is useful for the preventive treatment of eczema, asthma, allergic symptoms, allergic rhinitis and/or atopic disturbances, e.g. migraine, CrohnIs disease, ulcerative colitis, otitis media, nephrotic syndrcme and diabetes.

The composition of the inentiDn is particularly useful for the treatment of disturbances of the cardioascular sys4Le:T.: for a,-oPlectic rranifeúlatiol-s, thror.--c-e,-..bolic states suC,.

c) The the d) The 4? as stroke, infarction and Keshan ' s syndrome of young patients as well as for the Prevention of conditions.

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

The cA.;-3 polyunsaturated fatty acids used in the composition of the invention can be prepared according to Examples 1 and 2 whilst the algae enriched with selenium can be prepared according to Example 3. The pharmaceutical compositions according to the invention are described in Examples 4 to 6.

Ex2-Mie 1 2 kg of sodium hydroxide are dissolved in 70 litres of 95% ethanol- and 10 kg of cod-liver oil are added at a temperature of 50 to 60 OC. The mixtute is refluxed under nitrogen for 2 hours and then cooled to 10 0 C while stirring. The sodium salt of the saturated fatty acids are precipilCated. The crystals are filtered and washed with a little ethanol. The ethanolic filtrate is evaporated and 20 litres of boiled-out water are added to the residue. The nonhydrolyzable compounds such as cholesterol are completely removed by extraction with 5 litres of hexane. The aqueous phase is acidified to pH 2 by adding sulfuric acid and again extracted with 15 litres of hexane. The organic phase is washed with water, dried over anhydrous sodium sulfate and e.a,:ra"Led to give 3.2 kg of a concentrated oil with a DHA cortent of 36.8 % and EPA content of 31. 8 %. This oil is brown and smelling c,&" fish. Thus, it is mixed with Fuller's earth, heated under nitrogen /0 at 105 0C for 10 minutes and filtered as hot. The decdorizatici s a chi eved by st eam di st i 11 at i on under v@ cuum a t 1 -1 0 OC/ l Hgn, m for 3 hours to obtain 1.6 kg of a light yellow, tasteless, odourless oil with unchanged composition.

Example 2

8 kg of 40% sodium hydroxide solution are dropped at 50 to 60 OC to 24 kg of cod-liver oil dissolved at 60 OC in 16 litres of methanol under stirring. Then the mixture is stirred at 60 oC for additional 45 minutes. 20 kg of 15% hydrochloric acid are added to the solution at about 60 0C. After separation, the organic phase is washed with 1G kg of 15% hydrochloric acid and then with 180 litres of hot tap water until neutral. 'the phases are again separated, 100 litres of aetone are added to the oily phase which is then heated to about 4.5 OC and a solution of 3.8 kg of lithi-Jum hydroxide monchydrate in 30 litres of water are added. After stirring for 30 minlutes the mixture is left to stand overnight, then filtered and the acetone filtrate is evaporated. The residue is acidified by adding about 8 kg of 15% hydrochloric acid, extracted 3 times with hexane and evaporated. During the whole purifying operation nitrogen atmosphere is used. Thus, 6.4 kg of purified fish-oil are obtained with an iodine number of 258 and acid number of 160.

One kg of the cod-liver oil puPified as described is dropped at 60 OC to a solution containing 3 kg of urea in 9 litres of methanol. The mixture is stirred at the sare temperature for 2 hours and then cooled. It is left to t ) 1 at -10 OC overnight, then filtered and the filtrate is evaporated. 2. 5 litres of 1:1 hyd-ochlDric acid a7c added to the residue and the mixture is stirred for 15 minutes. After extraction with hexane, the hexane phase is washed with water until neutral, dried over anhydrous scdjum sulfatt. and evaporated to give 0.34 kg of W-3 unsaturated fatty acid with an iodine number of 315 which contains 24% of EPA and 42% of DHA.

Example 3 mg of sodium selenite are added to 8 litres of Knop-Pringsheimls culture medium filled in an alga-cultivating glass bottle of 10 litres volume. The thus-obtained culture medium is sterilized at 121 0 C under an overpressure of 1 bar for 30 miriutes. Then, the sterile solution is cooled and inoculated with a pure culture of Scenedesmus obtisiusculus which is capable to readily incorporate selenlurri. Sterile air containing 5% by volume of carbon dioxide is bubbled through the culture medium at 25 OLC while the system is illuminated by an electric discharge tube working with 4000 lux at a wavelength of 440 to 700 /um. After a cultivation period of 14 days the alga is separated from the culture medium. The thus-obtained alga mass is decomposed by supersound and carefully dried at a temperature below 65 OC. 1he yield i- 6 g of an alga powder with a selenium cor-,ent of 1200 1 :i /ugg- 121 Example 4

9 of a selenlum-containing alga powder (selenium content: 260 /ug/c) are added to 150 9 of. 65% enriched cod -liver oil (containing 22% of EPA and 43% of DHA). After homogenizing, 0.1% of vitamin E is added. The thus-obtained active ingredient is filled-into soft gelatine capsules and packaged into blister foils.

Example 5

The process described in Example 4 is followed, except that the following starting materials are used: 400 g of 65% enriched cod-liver oil with an EPA content of 22% and DHA content of 43%; 70,6 g of an alga powder with a selenium of 1200 /ug,/a; and 0.4 9 of vitamin E. The horrogenizate is filled into capsules capable of taking up 500 mg of active inGredient.

Example 6

Tablets with the following composition are prepared by using known pharmaceutical devices and processt Cod-liver oil enriched with.EPA and DHA and containing 0.1% of vitamin E (with a DHA content of 43% and EPA content of 22%) 200 mg Selenium-containing alga powder kselenium content:

1500 /ug/g) 86 mg .a-,".,cse 140 M9 C t 8:' c h 60 mg 13.

3. 5 mg 3.5 mg Poilvinlipyrrolidone Magnesium stearate if desired. the tailets map be coCed with sugar in a taoletting machine.

liJ- C 1 a 1 m!.

1. A pharmaceutical composition acting on the heart and cardiovascular system, which c c m p r i s e s 0.5 to 50% by mass of selenium-containing alga as well as 99.5 to 50% by mass of C 18-22 fatty acids containing at least two unsaturated bonds or the derivatives thereof, optionally in admixture with carriers and/or additives and antioxidants commonly used in the pharmaceutical industry. 1 4. A ccr,,position as claimed in claim 1, which c c m p r i s e s 5 to 35% by mass of selenium-containing alga.

3. A cc:ntposition as claimed in claim 1, which c c m p r i s e s 15 to 25% by mass of selenium-containing alga.

4. A composition as claimed in claim 1, which c 0 m p r i 5 E S 95 tO 65% by mass of C 18-22 fatty acids ccnta,;ning at least two unsaturated bonds or the derivatives thereof.

5. A composition. as claim ed in claim 1 which c c m p r i s e s 85 to 75% by mass of C 18-22 fatty acids containing at least two unsaturated bonds or the derivatives thereof.

6. A pharmaceutical composition as claimed in any of claims 1 to 5, which c c m p r i s e s fatty acids extracted from the oil of marine fishes as unsaturated fatty acids.

7. A pharmaceutical compositien as claimed in any of clairrs 1 to 6, which. c o r, p r i s e s 5,8,11,14,17-eicosapen4$aenc-ic ac"Ld and 's.,7. ,"0,13,16,19-dozosahexaenoic. acic as LnsaturatEd fatt acids.

4 i 8. A process for the preQe7ation of a pharmaceutical composition acting on the heart and cardiovascular system, which c o m p r i s e s mixing 0. 5 to 50% by mass of selenium-containing alga as well as 99.5 to 50% by mass of C 18-22 fatty acids containing at least two unsaturated bonds or the derivatives thereof optionally together with carriers and/or additives and antioxidants commonly used in the pharmaceutical industry.

9. A process as claimed in claim 8, in which 5 to 35% by mass of seleniumcontaining alga and 95 to 65% by mass of C 18-22 fatty acids containing at least two unsaturated bonds are admixed.

10. A process as claimed in claim 6, in.which 15 to 25% by mass of selenijm-containing alga and 85 to 75% by mass of C 18-22 fatt, acids containing at least two unsaturated bonds are admixed.

11. A pharmaceutical composition comprising selenium-ccntaining algae and one or more C 18-22 f atty acid(s) containing at least two unsaturated bondn or derivatives thereof, optionally in admixture with carriersand/or additives, e.g. antioxidants, commonly used in the pharmaceutical industry.

12. A composition as claiTed in claim 11 which includes, based on the combined weight of the selenium-containing algae and the c 28-22 fatty acid(s), or derivatives thereof, 0.5 to 50% of the selenium-containing algae and 50 to 99.5% of the C 18-22 fatty acid(s) or derivatives thereof.

13. A process of preparing a composition as claimed in claim 11 or 12, which comprises mixing together th- components thereof.

14. A composition as claimed in any one of claims 1 to 7, or 12 for use in treating heart or cardiovascular conditions.

14 15. Use of algae com.tined w-'tl,-j one cr r-,cre C, ratty acids containing at least two unsaturated bond. or L 8-22 " -1 de7Jvat--'ves thereof, in the preparation cf a r.e,-JJ_,zament fer treating heart or cardiovascular conditions.

16. A. composition substantially as hereinbefore described in any one of Examples 4 to 6.

17. A composition as claimed in claim 1 or claim 11, wherein the fatty acids and/or the algae are prepared by a process substantially as hereinbefore described in Example 1 or 2 or in Example 3, respectively.

18. A process as claimed in claim 13 substantially as hereinbefore described in any one of Examples 4 to 6.

1 Published 1988 a, The Pater. Ofice. State House. 6671 High Holborn. London WC1R 4TP. Further copies mkv be obtained from The Patent Oface. Sales Branch. St Mary Cray. Orpingwn, Kent BR5 3FLD. Printed by Multiplex techniques ltd. St Mary Cray, Kent Con. 1187-