FR2628972A1 - PHARMACEUTICAL COMPOSITIONS IN PARTICULAR IN THE CARDIAC AND CARDIOVASCULAR SYSTEM AND METHOD FOR THE PREPARATION THEREOF - Google Patents

Abstract

The invention relates to pharmaceutical compositions which act in particular on thromboembolic conditions of the cardiac and cardiovascular system, which comprise 0.5 to 50% by mass of a microorganism containing selenium as well as 99.5 to 50% by mass. mass of C 1 8 - 24 fatty acids containing at least two unsaturated bonds or their derivatives, optionally mixed with vehicles and / or additives commonly used in the pharmaceutical industry and optionally with antioxidants. Further, the invention relates to a process for preparing such compositions.

Description

() FRENCH REPUBLIC Publication No. 2,628,972 (to be used only

  for INSTITUTE NATIONAL Reproduction Orders)

OF THE PROPERTY = INDUSTRIAL TE

  OF INDUSTRIAL PROPERTY N of national registration 89 03812 PARIS

  () Int CI'4: A 61 K 33/04, 31/20, 35/74.

@ APPLICATION FOR PATENT OF INVENTION A1

  ) Date of deposit: 23 March 1989. Q Applicant (s): BIOREX KFT - HU.

  ) Priority: HU, 23 March 1988, No. 1484/88.

  (Inventor (s): Péter Literati Nagy: Erzsebet Radvany born Hegedus: Maria Boros, Jeno Szilbereky, Gyorgy) Date of making the Blask6 available to the public; Bela Gachilyi; Gizella Hamar born Nemes; Gabor

  application: BOPI "Patents" No. 39 of 29 September 1989. Nemeth.

  References to other national documents appearing

rented: rn Owner (s):

  Attorney (s): Cabinet Ores.

  Pharmaceutical compositions acting in particular on the cardiac and cardiovascular system and process for

prepare them.

  The invention relates to pharmaceutical compositions which in particular act on the thromboembolic states of the cardiac and cardiovascular system, which comprise 0.5 to% by weight of a microorganism containing selenium as well as 99.5 to 50%. in bulk of C18.24 fatty acids containing at least two unsaturated bonds or their derivatives, optionally mixed with vehicles and / or additives commonly used in the pharmaceutical industry and

  possibly with antioxidants.

  In addition, the invention relates to a process for preparing these compositions. I 1% N IL N Co UL Sales of fascicles to NATIONAL PRINTING. 27. rue de la Convention - 75732 PARIS CEDEX 15

  The present invention relates to pharmaceutical compositions

  Ceutiquei acting in particular on the thrombo-embolic states of

  cardiac and cardiovascular system.

  - According to another aspect of the invention a proposal is

  assigned for the preparation of said compositions. It is known that the w-3 unsaturated C18-24 fatty acids have advantageous biological properties. Among these compounds, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are remarkable. Dyerberg et al. / The Lancet 15, 117 (1978) 7 emphasized the importance and the multiple biological effects of these two acids.

  The effects related to the important role of poly-

  saturated8 in particular EPA and DHAq in hyperlipidemia and

  thrombotic conditions have been reviewed by Goodnight et al.

  / fArteriosclerosis 2, 87 (1982) Review 7.

  Pharmaceutical compositions containing EPA and DHA as active ingredients have been described, for example,

  in the specification of German Patent No. 3,438,630 for the reduction of

  of blood cholesterol, and further in the Japanese patent applications published under Nos. 58.08037 and 60. 49097 against cerebral sclerosis and to prevent thrombus formation in

  patients with heart disease.

  Several articles have been devoted to the inhibitory effect of aggregation piaquettaires thus the inhibitory effect of formation of the

  thrombus of EPA and DHA i Spencer and Caraegas Prostagl. Leucotrie-

  r.es and Med. 23, 129 (1986); Knapp et al. New Engl. Journ. of Med.

314,937 (1986) 7.

  The antiviral action of EPA and DHA is described in

  U.S. Patent Specification No. 0 4,513,008.

  The active ingredients of fish oil, for example

  EPA and DHA are precursors to the biosynthesis of

  PG-3 and they inhibit the formation of harmful metabolites such as TXA2 and TXB2 prover.nant of the "arachidonic acid cascade", so-called that is a catech of complicated biochemical processes starting

arachidonic acid.

  Apart from their various favorable effects, the polyunsaturated fatty acids have the disadvantageous property of being subjected to spontaneous decoemposition by oxidation in the human body, thus giving rise to the formation of active aldehydes, such as malonodialdehyde which is harmful to the body. These aldehydes are able to react primarily with the connective tissues of

  a physiologically harmful way that can lead to "lipofus-

  the so-called central nervous system of the central nervous system.

  Research has begun on the biological effects of microelements and trace elements in the last decade. It is now known that selenium is one of the most important and indispensable substances for life. The favorable action of selenium is mainly due to its activating effect towards

  of the enzyme glutathione peroxidase because selenium is a

  indispensable tituart of the prosthetic group of the enzyme glutathione-

  peroxidase. This enzyme is the most important endogenous inhibitor

  pernicious peroxidation processes.

  Selenium in itself has a hypotensive, ameliorative effect.

  re ischemic, hypoxic and infarctic states of the heart and inhibits

  lipofuscinose ceroldep and in others it also has an

  tageux on the periodontite. It has significant anticancer activity and decreases the likelihood of cancerous disease progression. Furthermore,

  it is considered as an inhibitory agent for mutagenesis.

  Selenium does not accumulate in the body, so well

  that it must be supplemented continuously. Until then, selenium has been

  usually introduced into the body in the form of inorganic compounds

  for example, selenium dioxide and sodium selenite. A number of alterations or pathological conditions such as

  hepatic necrosis muscular necrosis the destruction of the

  Erythrocyte bran intersticially lesions ST elevation in ECGs Mulberry heart syndrome, kwashiorkor syndrome

  (protein malnutrition) and multiple sclerosis were found to be

  respectively triggered by selenium deficiency.

  A review of the overall biological effects of selenium has been published by Thressa et al. / Nutrition Review 35 * 7 (1977) 7,

  Shamberger / = J. of Env. Path. and Tox. 4s 305 (1980) 7 and Masum

  kawa and coil. Experientia 39, 405 (1983) 7.

  The organic selenium compounds can be obtained by synthetic methods described, for example, by Klaiman and Gunther in their work entitled "Organic Selenium Compounds, Their Chemistry and Biology", John Wiley and Sors., Inc. ew-York. , 1973. By

  In addition, it is also possible to prepare the organic compounds of the

  using microorganisms. According to this last process

  cultivates microorganisms on media enriched with inor-

  selenium. Suitable organisms for this purpose bind selenium and, by introducing it into their metabolism, they bind it to organic substances, most frequently to amino acids.

  or fats. These possibilities are described by Danch and Clbmielow.

  sky./ Pr. Nauk. Univ. Slask Katowich 57 (1985) 7 and by

  ty et al. / Biochem. Biophys. Res. Commumn. 118, 176 (1984) 7.

  A number of microorganisms are known that are able to collect some elements, such as selenium, in

  their environment and to accumulate them in their own organism.

  The yeast preparation enriched with selenium is de-

  disclosed in U.S. Patent Specification No. 4,530,846 and in Japanese Patent Applications Nos. 60.224451 and 57.174098.

  The enrichment of selenium by other microcracks

  Nismess such as actinomycete and hyphomycete bacteriins, besides yeasts is described in the Japanese Patent Application published under

N 78.148587.

  According to the Japanese patent application published under No. 58.129954, the use of yeast containing selenium is proposed,

  mixed with vegetable oils to treat serological diseases

  the. The object of the present invention is to provide a new pharmaceutical composition acting on the cardiac and cardiovascular system which makes possible the combination of the properties

  advantageous microorganisms containing organic selenium

  with DHA and EPA, and simultaneously eliminates

  disadvantageous properties of polyunsaturated fatty acids.

  The invention is based on the discovery that the aforementioned objective can be fully achieved through useful compositions

  to eliminate lipofuscinose ceroid, which can be obtained using

  By virtue of a feature of the invention, a microorganism containing selenium which is toxic from the point of view of human use, in conjunction with polyunsaturated fatty acids, is thus provided with a pharmaceutical composition acting on the cardiac system.

  cardiovascular and which comprises 0.5 to 907. in mass of a microorganism

  containing selenium and 9995 to 50% by mass of fatty acids

  at C18, 24 with at least two unsaturated bonds and / or their dice.

  riveted, optionally in admixture with vehicles and / or additives commonly used in the pharmaceutical industry and possibly

with antioxidants.

  In addition, the invention is directed to a process for preparing the aforesaid composition, which comprises mixing 0.5 to 50

  mass of a microorganism containing selenium as well as 99.5 to 50 /.

  mass of C18.24 fatty acids containing at least two

  saturated and / or their derivatives, optionally with vehicles and / or additives, commonly used in the pharmaceutical industry and possibly with antioxidants, in a manner commonly known

in the pharmaceutical industry.

  The composition according to the invention comprises suitably

  eicosapentaenoic acid (EPA) and docosahexaenoic acid

  (DHA) as well as a yeast containing selenium.

  As raw materials of C 3 -unsaturated fatty acids

  Res C18.24 represents one of the components of the composition of the invention, can be used oils that can be obtained from various marine fish and freshwater mainly from mackerel, herring cod sardine, or molluscs such as squid as well as from the liver of said fish, for example

  cod liver oil and shark liver oil.

  In addition to EPA and DHA, fish oils

  hold a large amount of saturated fatty acids as well as

  partially unsaturated fats and other non-hydrolytic

  sands. The elimination of these constituents is very important if the administration of the therapeutic compositions prepared from fish oils would have the effect of substantially increasing

  the amount of calories introduced and the blood triglyceride level

  wrinkles. In addition, non-hydrolysable constituents may contain

  steroids such as cholesterol, vitamin D (and its provita-

  mine) and / ordelavitamin A. Vitamins D and A are accumulated in the human body so that prolonged treatment, necessary for the desired effect would be impossible using a composition

  containing these vitamins. As a result, we must eliminate from the oil

  the above-mentioned constituents, such as fatty acids,

  partially unsaturated and non-hydrous constituents

  lysabless eg cholesterol and vitamins A and D. In this way the total amount of EPA and DHA of the fish oil is

more than 50% enriched.

  It is appropriate to use micro-organism strains

  which are not toxic to the human organism and are useful for human consumption. Genes widely used in biotechnology, such as Lactobacillus Leukonostocp Pediococcus,

  Acetobacter, Streptococcus Torula, Kluyveromycess Candida, Brettano-

  myces, Brevibacteriums Saccharomyces, Torulopsis, Pichia, Hansenula,

  Oidiums Rhodotorula "Trichospora, Penicillitum Rhizopuss Mucors Monas-

  cuss Aspergilluss among others # that are useful for consumption

  Human O20s are very suitable for this purpose.

  To inhibit the oxidation of the composition of the invention,

  tion, it is appropriate to use "(-tocopherol (vitamin E),

  glutathione or common antioxidants such as butylhydroxy.

  toluene as active preservatives.

  It is possible to use substances or carriers as substances.

  these commonly used in the pharmaceutical industry, such as lactose starch or magnesium stearate,

  Pharmaceutical research has shown that the compound

  The invention is devoid of damaging side effects.

  because there is no change in the micro-system.

  so-called enzymatic liver in rats using a hundred times the amount

  the current dose. The amount of lipofuscin accumulated in

  the organism is significantly lowered compared with the witness a-

  closely with the composition of the invention. According to examinations carried out using the composition of the invention in

  Wistar rats for 6 weeks # an inhibitory effect of

  platelet aggregation free from ambiguities.

  The optimum daily effective dose (calculated for an average body weight of 70 kg) of the composition of the invention is 0.1 to 5 g. The oil component contains 22% EPA and 43% DHA on average. The effective daily dose is 100-300 μg of selenium.

  The main advantages of the ccmposition of the invention

  can be summarized as follows: 1. The advantageous properties of EPA and DHA as well as

  that those of selenium and yeast are combined.

  2. The harmful effects of known compositions

  containing fish oils which are caused by

  such as sterols and vitamins A and Ds are

clouds swept away.

  3.The possibility of occurrence of "Iipofuscinosis"

  The composition of the invention contains selenium as an incipient substance enriched in a micro-organism such as in the consumption of polyunsaturated fatty acids.

  only bacteria, fungi or yeasts. Selenium administered jointly

  with the specific micro-organism is better absorbed and exerts

  preferably, its favorable effect.

  5. The selenium bound in organic compounds can

  Be economically prepared in large quantities, specifically

  richi in microorganisms, mainly in yeasts.

  6. The composition exerts a therapeutic action promoting

  in the case of atopic disorders and is useful for the preventive treatment of eczema, asthma, allergic symptoms, allergic rhinitis and / or atopic disorders such as migraine.

  Crohn's disease # ulcerative colitis, otitis media nephrotic syndrome

  that and diabetes. It is particularly useful for the treatment of

  disorders of the cardiovascular system: for apoplexy

  ticks, thromboembolic states such as infarction and syndromes

  of Keshan in young patients, as well as for prevention

abnormal conditions.

  The invention is illustrated in detail by the Examples

  following, given as an indication, but in no way limiting.

  The polyunsaturated fatty acids used in the

  composition of the invention may be prepared in accordance with the

  pie 1, while the selenium-enriched microorganism can be

  prepared as described in Examples 2 to 6. The pharmaceutical compositions are

  The ticks according to the invention are illustrated by Examples 7 to 9.

Example 1

  8 kg of a 40% sodium hydroxide solution at 50-600C are poured into 24 kg of mackerel oil dissolved

  600C in 16 liters of methanol with stirring. Then we shake the mixture

  at 600C for an additional 45 minutes. After addition of 20 kg of 15% hydrochloric acid to the solution at about 60 ° C., the mixture is separated.

  the layers and the organic phase is washed with 10 kg of hydrochloric acid

  dries at 15%. then with 180 liters of hot tap water, until neutral. The phases are separated again, 10C liters of acetone are added to the oily phase which is then heated to approximately 450 ° C. and 3.8 kg of lithium hydroxide dissolved in 30 liters of calcium are added.

  water. After stirring for 30 minutes, the mixture is allowed to stand.

  overnight, then filtered and the filtrate evaporated.

  tonic. The residue is acidified by adding about 8 kg of 15% hydrochloric acid, extracted 3 times with hexane and the mixture is evaporated.

  hexane solution. An atmosphere of nitrogen is used during the operation

  entire purification. 6.4 kg of pea oil are thus obtained.

its purified.

  1 kg of the purified mackerel oil as described above is poured dropwise at 600 ° C. into a solution containing 3 kg of urea in 9 liters of methanol. The mixture is stirred at the same temperature for 2 hours and then cooled. We leave him at

  at -10 ° C overnight, then filter and evaporate the

  trat. 2s5 liters of 1: 1 hydrochloric acid are added to the residue and the mixture is stirred for 15 minutes. After extraction with hexanes, the hexane phase is washed with water until neutral, dried over anhydrous sodium sulphate. and evaporated to 0.34 kg of W-3 unsaturated fatty acids having an iodine number of 315 and containing 247.

EPA and 42% DHA.

Example 2

  Each time, 2 liters of culture medium are poured into sterilized liquid malt extract in glass flasks of a volume of 5 liters and the medium is enriched by adding 3% by weight of glucose. After adding 5 g / wl of sodium selenite, it is

  inoculates with Saccharomyces cerevisiae. then it is grown under aeration

  & 320C for 72 hours. An additional 51ug / ml of sodium selenite is added to the culture at the 48th hour of the day.

culture operation.

  After filtration, the cells are washed several times

  with water and dried at 68-70 C. After micronization,

  terminates the selenium content by atomic absorption by obtaining a

value of 2600P g / g.

  EXAMPLE 3 100 ml of a culture medium is poured into the liquid yeast extract in Erlenmeyer flasks with a volume of

  500 ml under sterile conditions. After addition of 5 μg / ml of

  Sodium lenite is inoculated with Aspergillus soy mildew and shaken cultures at 28-30 C. An additional amount of 5 g / ml of sodium selenite is added to the culture the 3rd

  day of the culture operation. After 5 days, the material is collected

  It is dried by filtration, washed and dried as described in Example 2. After micronization, the selenium content of a sample is determined by atomic absorption and found to be 3000 μg / g.

Example 4

  The procedure described in Example 3p is followed except that the yeast Torulopsis used is used instead of mold. The microbial mass obtained during the cultivation

1200 g / g of selenium.

Example 5

  Streptococcus thermophilus is cultured at 45oC in a

  culture medium prepared as described in Example 3.

  subsequent treatment and treatment the microbial powder contains 4200

pg / g of selenium.

Example 6

  The bacterium Lactobacillus mesenteroides

  described in Example 39 except that the medium is supplemented by the addition of 1% glucose and 2% calcium carbonate by obtaining

  a bacterial mass containing 3000ug / g of selenium.

Example 7

  10 g of a yeast powder containing

  lenium (selenium content: 2600 / ug / g) to 150 g of mackerel oil

  (containing 24% EPA and 42% DHA) enriched to 65%. After homogeneity

  The mixture is preserved by the addition of 0915 g of vitamin E. The active agent thus obtained is then mixed with gelatin capsules.

  soft that lion packs into blister sheets.

  EXAMPLE 8 The procedure described in Example 7 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 a DHA content 43%; 100 g of yeast powder with selenium content of 1200 pug / g; and 0 & g of vitamin E. Capsules capable of receiving 500 mg of active agent are each filled with 1 homogenizate.

Example 9

  The procedure described in Example 8 is followed except that 400 g dl is used as a fish oil component.

  a fish oil at 30 /. (containing 18% EPA and 12% DHA)

  by Martens and Co., Bergen, Norway).

Example 10

  The procedure described in Example 8 is followed, except that 400 g of fish oil component is used.

  fish oil containing 9.2% EPA and 10.4% DHA.

Example 11

  Tablets having the following composition are prepared using known pharmaceutical devices and methods: EPA and DHA-enriched cod liver oil containing 011% vitamin E (at DHA content 437 and EPA content 22) %) 200.0 mg Yeast powder containing selenium (selenium content: 1500 ug / g) 86 $ 0 mg Lactose 140.0 mg Starch 60.0 mE Polyvinyl pyrrolidone 3.5 mg Magnesium stearate 3.5 mg Si it is desired, the tablets can be coated with sugar or with another substance in a machine to be made

tablets.

  It is understood that lion can implement the invention according to many variations and modifications, without

  as much to move away from his frame and his mind.

Claims (10)

  1. Pharmaceutical compositions acting on the system   cardiac and cardiovascular systems characterized in that they comprise   0 to 50% by weight of a microorganism containing selenum and 99 to 50% by weight of C 18-24 fatty acids containing at least two   unsaturated bonds and / or their derivatives, optionally in admixture with   vehicles and / or additives commonly used in industry   pharmaceutical and possibly with antioxidants.   2. Composition according to claim 11, characterized in that it comprises 5 to 357. en masse of the microorganism containing selenium and 95 to 65% by weight of unsaturated fatty acids or their derivatives. 3. Composition according to claim 11, characterized in that it comprises 15 to 25% by weight of the microorganism containing   selenium and 85 to 75% by weight unsaturated fatty acids.   4. Composition according to claim 1, characterized in that it comprises fatty acids extracted from fish oil   as unsaturated fatty acids.   5. Composition according to claim 1 characterized in that it comprises 5s81114,17-eicosapentaenoic acid and 1 '   acid 497 $ 10s13,16,19-docosahexaér.olque as unsaturated fatty acids tured.   6. Process for the preparation of a pharmaceutical composition   heart and cardiovascular system, characterized by   in which 0.5 to 50% by weight of a microorganism containing   selenium as well as 9995 to 50% by weight of C18.24 fatty acids containing at least two unsaturated bonds or derivatives thereof with   vehicles and / or additives commonly used in the pharmaceutical industry.   and possibly with antioxidants of about one   known in the pharmaceutical industry.   7. Method according to claim 6p characterized in that   5 to 35% by weight of the microorganism containing   with 95 to 65% by mass of unsaturated fatty acids.   8. Process according to claim 6, characterized in that   15 to 25% by weight of the microorganism containing   with 85 to 75% by mass of unsaturated fatty acids.   9. Process according to claim 6, characterized in that fatty acids extracted from marine fish oil are used. as unsaturated fatty acids.   10. Process according to claim 6, characterized in   what it includes the use of 5s8,1114p17-eicosapenta acid   enolque and 4s7O10,13,16,19-docosahexaenoic acid as acids unsaturated fat.