DE3438630A1 - COMBINED FATTY ACID COMPOSITION FOR LOWERING CHOLESTERIN AND TRIGLYCERIDE BLOOD LEVEL - Google Patents

Description

■ ^: - "■""' ^: 3A38630

description

The invention relates to pharmaceutical compositions and foodstuffs containing a specific combination in fatty acids. These pharmaceutical compositions Tongues and foods can be used to treat humans or other mammals with the aim of lowering the Blood cholesterol and triglyceride levels can be used.

It is known that atherosclerotic cardiovascular disease is rare in Greenland Eskimos. This has been attributed to the fact that Eskimos ingest large amounts of fish oil. The active ingredients of fish oil are (throughout-Z) -5,8,11,14,17-eicosapentaenoic acid, sometimes referred to as 20: 5eJ 3-fatty acid (referred to herein as "EPA") and (throughout -Z) -4, 7,10,13,16,19-docosahexaenoic acid, sometimes as 22: 6 (ύ 3 fatty acid (hereinafter referred to as "DHA"). EPA and DHA are known precursors in the biosynthesis of prostaglandin PGE.,.

The above alternative designations, such as 20: 5w3, give © in front of the colon the number of carbon atoms in the chain; after the colon the number of double bonds; and after the omega, the number of carbon atoms at which the first double bond occurs, from the end opposite the carboxylic acid group, members of a certain omega series of fatty acids, such as CO 3, can usually be converted into acids with different lengths and different total numbers of double bonds by normal body enzymes However, in general, a transition from one compound from one Omega row to another, e.g. (ύ 3 to W 6, is not possible. This is due to the fact that body enzymes generally change the length and the double bonds of the carbon

- cause 6 * acid end of the chain.

In GB-PS 1 604 554 and 2 033 745 it is described that EPA can be used as an effective agent for the therapy and prophylaxis of thromboembolic conditions such as myocardial infarction, strokes or deep-seated vein thromboses ¹ in the course of surgical interventions. These publications describe the extraction of EPA from fish oil, such as cod liver oil or menhaden oil. EPA can be given instead of butter or normal margarine in the form of a special margarine that is formulated in such a way that the required amount of EPA is absorbed during normal consumption.

This practice has, in spite of the fact, that it is natural, easily incorporated into the daily diet Substance served, not yet found widespread. One reason for this may be that it difficult "is to get a reasonable price EPA from fish oils separate to form a pure product. Another reason may be that the administration EPA does not have as drastic an impact as was assumed.

The object of the invention is to overcome the disadvantages of the prior art explained above, and in particular the compositions as disclosed in GB-PS 1604 and 2,033,745. In this context, compositions are to be provided which, in comparison

O to conventional products a superior therapeutic Have an effect. These therapeutically effective combinations

. Settlements are supposed to be naturally available · fatty acids which are used to lower blood levels of cholesterol and triglycerides. After all, they should therapeutic compositions of the invention increase PGE. : PGEp ratio in the patient and one Increase in the absolute amount of PGE. effect in the system.

This object is achieved according to the invention by the simultaneous administration of EPA and / or DHA together with one or more components from the group dihomo-if-linolenic acid (8,11,14-eicosatrienoic acid), ie 20:36) 6-fatty acid (hereinafter referred to as DHLA referred to), cis-linoleic acid ((Z, Z) -9,12-octadecadienoic acid), ie 18: 2 iü 6-fatty acid, and / "- linolenic acid ((Z, Z, Z-6,9,12-octadecatrienoic acid) , ie 18: 3W 6 fatty acid, either in the form of a pharmaceutical preparation, in the form of a food such as margarine or edible oil, or ¹ ^ dissolved in the form of a skin ointment or lotion for topical use.

The prostaglandins are a family of substances that have a variety of different biological effects demonstrate. Prostaglandins of the 1, 2 and 3 series have 1, 2 or 3 double bonds in the basic structure of a carboxylic acid with 20 carbon atoms, which comprises a 5-membered cyclopentene ring.

The 1 series of prostaglandins are powerful vasodilators Agents and inhibit the biosynthesis of cholesterol and collagen, as well as platelet aggregation. The prostaglandins of the 2 series strengthen platelet aggregation, the biosynthesis of cholesterol and collagen and also effect an increased proliferation of endothelial cells. The main effect of the prostaglandins of the 3 series, in particular from PGE, is a repression of the prostaglandins of the 2 series.

The precursor to the 2-series prostaglandins is arachidonic acid ((throughout Z) -5,8,11,14-eicosatetraenoic acid), i. 20: 4io 6 fatty acid. DHLA is the forerunner for that. Prostaglandins of the 1-series and EPA and DHA are, as explained above, the precursors of the prostaglandins of the 3-series.

It is believed that the effectiveness of EPA and DHA in preventing ^ atherosclerotic cardiovascular Diseases both on their effect as precursors for pro-

staglandin PGE ,,, which promotes the formation of the prostaglandins of the 2 series suppressed, as well as based on the fact that EPA and / or DHA itself is the same with arachidonic acid compete enzymatic system and thus inhibit the biosynthesis of the prostaglandins of the 2 series. This inhibition of Prostaglandins of the 2 series leads to an increase in the ratio of PGE.:PGEp.

In order to improve the effect of the administration of EPA and / or DHA all.ein, with the aim of _{further increasing the PGE 1} : PGEp ratio and ensuring an increase in the absolute amount of PGE ₁ in the system, DHLA should simultaneously can be administered with the pure EPA and / or DHA. Since cis-linoleic acid and ^ -linolenic acid both form DHLA in the body's metabolism, DHLA can be replaced in whole or in part by one or both of these fatty acids.

It has been found that the combination of EPA (and / or DHA), and DHLA (and / or cis-linolenic acid and / or ^ "-linolenic acid) causes a substantial reduction in cholesterol and triglycerides in the blood. Recent research has finally confirmed that that there is a connection between the level of cholesterol in the blood and the occurrence of coronary heart disease; JAMA, vol. 251 (T $ 84) _f pp. 351-364 and JAMA, vol. 251, pp. 365-374 (1984). Further, it is expected that a _s such combination further beneficial therapeutic properties. for example, it is known that in schizophrenia, rheumatoid arthritis and other collagen and autoimmune diseases as well as in some forms of cancer indications for extremely low PGE ^ concentrations and high PGEP concentrations exist. It is therefore to be expected that the combination according to the invention will contribute to an effective treatment of such conditions. Furthermore, it is assumed that the anti-inflammatory effect of Co rticosteroids and the analgesic effect of acetylsalicylic acid (aspirin) is due to the repression of PGE ₂ formation. It can thus be expected that the combination according to the invention will be a natural and very effective

represents a common anti-inflammatory, pain reliever.

The dose of the composition according to the invention, the one Combination of EPA (and / or DHA) and DHLA (and / or cis-linoleic acid and / or ^ -linolenic acid), which for a therapeutic or prophylactic effect required varies depending on the route of administration and the nature of the condition to be treated, but is generally at least 1 g and preferably 1.5 to 3 g per day. This is the dose for an average patient with a body weight of 70 kg. The dose for other patients or animals can accordingly depend on the weight vary, i.e. about 20 to 40 mg / kg. ■

·.

The relative amounts of EPA (and / or DHA) and DHLA (and / or cis-linoleic acid and / or / or "- linolenic acid) are in the Composition preferably 1: 1, this ratio can vary from 3: 1 to 1: 3.

EPA (and / or DHA) and DHLA (and / or cis-linoleic acid and / or ^ -linolenic acid) do not need to be administered as the acids themselves but can be used in the form of their pharmaceutically acceptable salts, esters or amides.

Esters or amides that are in vivo in the acid and other pharmaceutical compatible products can be converted can be used. The preferred ester is the ethyl ester. The preferred salts are the sodium and potassium salts. Any other pharmaceutically acceptable ones also come solid salts in question, provided they are suitable for tableting.

The inventive composition is preferably on administered orally, as this is the most convenient route of administration represents. However, the active ingredients can also be used in any other way in which successful absorption administered, e.g., parenterally (i.e. subcutaneously, intramuscularly or intravenously), rectally or

vaginally and topically, e.g. in the form of skin ointments or lotions.

It is possible to administer the active ingredients as such in the form of a simple mixture of the individual components. However, they are preferably administered in the form of pharmaceutical preparations. The preparations according to the invention for ' Veterinary and human medical purposes comprise the active ingredients defined above together with one or.

several pharmaceutically acceptable carriers for this and, if necessary, further therapeutic ingredients, but other unsaturated fatty acids, in particular arachidonic acid, should be avoided if possible. The carrier (s) must be pharmaceutically acceptable, ie they must be compatible with the other components of the preparations and must not have any adverse effects for the recipient. These preparations include those which are suitable (including subcutaneous, intramuscular and intravenous) for oral, rectal, vaginal, intrapulmonary, or _t parenteral administration. Preparations for oral administration such as tablets or capsules are preferred.

The combination of EPA (and / or DHA) and DHLA (and / or cis-linoleic acid and / or ^ -linolenic acid) can also be administered by mixing butter and / or normal margarine with a special margarine, e.g. of the emulsion type, the is composed in such a way that, with normal use by the consumer, the required amount of the combination is absorbed is replaced. Edible oils and fats can be provided in a similar manner so that they have the inventive Composition included.

EPA (and / or DHA) and DHLA (and / or cis-linoleic acid and / or / or -linolenic acid), which are used in the compositions according to the invention should be as pure as possible. EPA and / or DHA cannot be used directly in the form of fish oil because when using fish oil in the amount

which provides the desired amount of EPA and / or DHA, too many calories and potentially toxic amounts of the Vitamins A and D are provided. Thus, pure EPA and / or DHA should be extracted from the fish oil. The in-entity of other unsaturated fatty acids besides EPA, DHA, DHLA, cis-linoleic acid and / '- linolenic acid should be avoided will.

Substantially pure EPA and / or DHA can be extracted from fish oil, such as cod liver oil, by a method such as that described in US Pat. No. 4,377,526. Another possibility for their separation consists in a new procedure, which is also the subject of the invention, in which the double bonds of the unsaturated fatty acids in the fat or oil used as the starting product are iodized. These iodization allows protection of fatty acids from oxidation during processing and the resolution increases the fatty acids in the later column _t nchromatographie. After iodination, the fat or oil is saponified. The iodized fatty acids are extracted from the saponified mixture, then methylated and separated by column chromatography. The desired fractions are then de-iodized. This process can be used not only for the separation of EPA and DHA from fish oils, but also for the separation and extraction of other unsaturated fatty acids such as linolenic acid and linolenic acid; from the triglyceride forms in which they occur naturally, for example in soybean oil, linseed oil, safflower oil, oil from evening primrose and the like, are used. The separation of unsaturated fatty acids can be facilitated by means of the iodination process according to the invention.

The starting material of the process according to the invention can be natural fats or fatty oils, the The first stage is iodination, followed by saponification. However, this can also be the case with the starting material Any mixture of unsaturated fatty acids that are difficult to separate. In this case there is

the first stage in iodination, but no saponification is required, as this is the case with the starting material is not about triglycerides.

Iodination takes place by organizing iodine in one Slowly add solvent, preferably in the form of a 20 percent solution in ethanol, to the starting material, until the color in the starting material no longer disappears. This reaction takes place at room temperature under constant. Stirring instead.

The saponification step can be carried out in a conventional manner for example, by treatment with a 20 percent solution of KOH in ethanol for 2 hours.

The iodized fatty acids are obtained from the saponification mixture by conventional methods, for example by extraction with ether, extracted.

The next step in the process is the methylation of the iodized fatty acids to prepare them for column chromatography. Again, it is a conventional one Stage, which can be carried out, for example, with a 5 percent solution of hydrogen chloride in methanol can.

Finally the fatty acids are separated by column chromatography. Column chromatography is based on conventional Manner carried out with a conventional elution mixture.

While the separation of the various fatty acids is very poor in conventional processes, this separation is greatly improved when the fatty acids in column chromatography Treatment are iodized. The column can be packed with silica gel in a conventional manner. as Elution solutions can be any conventional solution such as hexane / ether / acetic acid (85: 10: 5).

After separation into fractions by column chromatography

the fatty acids are de-iodized, for example using of silver nitrate.

Although above for the various stages of the invention Process specific reagents and process conditions have been given, it is for those skilled in the art clear that other reagents and process conditions "are required to carry out the individual process steps desired are suitable. The critical factor is iodination prior to chromatography to achieve separation or dissolution to increase and protect the fatty acids against oxidation.

As described above, the separation is carried out by column chromatography. It should be pointed out, however, that other separation processes are also possible, for example high-speed centrifugation. The separation is improved also in such other 'separation process by the _t iodination.

The following example serves to explain the inventive Process for the separation of EPA and DHA from cod liver oil.

Manufacturing example

A ^ 20 percent solution of iodine in ethanol will be slow added to 300 g of cod liver oil. The addition of iodine is continued until the iodine color in the oil disappears. The reaction takes place at room temperature with continuous stirring. After iodination is finished, the iodized oily solution by treating with a 20-. percent solution of KOH in ethanol saponified. The iodized Fatty acids (260 g) can be easily removed from the saponification

Extract mixture.
35

The iodinated fatty acids are then methylated with a 5 percent solution of hydrogen chloride in methanol. EPA and DHA are column chromatography (1500 g silica gel,

3A38630

70 to 230 mesh, Merck). The elution is carried out with 5 liters of hexane / ether / acetic acid (85: 10: 5). at the first fraction extracted is iodized DHA. The second fraction is iodized EPA.

The essentially pure methylated and iodinated ' The fatty acid mixture can also be prepared by other conventional procedures, for example by high speed centrifugation or distillation. The de-iodination will performed by separating the iodized Me-DHA and Me-EPA with a 10 percent aqueous silver nitrate solution shakes. A silver jo'did precipitate forms. The organic phases are separated off. Same procedure is repeated until no more precipitation occurs. Microanalysis, HPLC and NMR show that the desired Products have been received. The yield is over 90 percent and the degree of purity is "96 to 100 percent. It is not necessary to carry out the process under nitrogen, since the fatty acids are saturated with iodine, whereby oxidation is prevented.

The following clinical investigations explain the synergistic effect that results when the inventive Combination achieves wind. For comparison, the effects are given when the individual components are administered alone specified.

Clinical examination

36 male and female outpatients * aged

35 to 75 years are divided into three groups of 12 Pa- each. divided into clients. The individual groups receive 45 days long in addition to your normal diet 5 cm / day free fat

• 3 acids. For Group I, the additive consists of 5 cm / day of essentially pure EPA. For group II the diet becomes with 5 cm / day essentially pure cis-linolenic acid and in group III with 3 cm / day essentially pure EPA and 2 cm / day of essentially pure cis-linoleic acid added.

1 By the term "substantially pure" is meant a purity of about 96 to 100 percent.

Cholesterol and triglycerides in the blood are tested 1 day before the start of treatment and after the 45-day treatment. The treatment results are summarized in Tables I, II and III below.

Al
ter Table I: 250 5 cm "/ day pure EPA Trigly
ceride
after 45
Days
mg / 100 ml Pa
tient
No. 45 Total-
chole-
Sterine
bad 1 day
before the
Treat
lung,
mg / 100 ml 248 Total-
chole-
sterol
after 45
Days
mg / 100 ml Trigly
ceride
1 day
before the
Treat
lung,
mg / 100 ml 102 1 45 M. 230 220 130 95 2 47 M. 270 220 115 90 3 73 M. 280 210 102 90 4th 60 . M. 265 240 95 95 5 56 W. 215 240 ■ 115 105 6th 54 M. 285 260 120 90 7th 52 W. 300 205 95 100 8th 63 W. 350 250 115 95 9 64 M. 190 250 110. 105 10 55 M. 200 260 • 160 80 ' 11 ν 35 M. 256.9 + 43.2 190 95 90 12th Average M. 190 90 94.95 + 7 227.9 + 24.4 111.8 + 18.6 15% % Decrease 11.2%

age Tabel - - 16 _ 3438630 ; % Decrease 190 Total-
chole-
sterol
after 45
Days
mg / 100 ml Trigly
ceride
1 day
before the
Treat
lung,
mg / 100 ml Trigly
ceride
after 45
Days
mg / 100. ml 47 II: 5 cnr / day of pure cis-linoleic acid 350 200 95 95 75 Total-
Chilled
bad sterol
1 day
before the
Treat
lung,
mg / 100 ml 200 340 90 95 * Pa
tient
No. 60 M. 220 205 110 105 1 60 W. 240, 220 115 100 2 55 W. 270 • 210 ■ 90 100 3 37 W. 220 260 105 95 4th 40 W. 400 230 80 80 5 45 M. 310 350 Ί65 150 6th 62 M. 230 300 140 140 7th 54 M. 260 220 ■ . 115 115 8th 52 W. 215 250 110 110 9 61 ' M. 265.3-61.75 215 130 125 10 Average M. 250 ^ 50 112 ^ 22.1 109, i6ii9.4 11th W. 5% 2% 12th

Table III age : 3 cm ³ '"" -17' % Decrease 450 ... - 3438630 Trigly
ceride
after 45
Days
mg / 100 ml Pa
tient
No. 48 / Day pure EPA 310 - pure cis-linoleic acid 90 1 1 60 Total-
Chilled
bad sterol
1 day
before the
Treat
lung,
mg / 100 ml 257 and 2 cdI / day Trigly
ceride
1 day
before the
Treat
lung,
mg / 100 ml 40- 5 2 45 M. 305 Total-
chole-
sterol
after 45.
Days,.
mg / 100 ml 160 50 3 40 M. 210 ' 260 70 45 10 4th 54 M. 210 240 106 55 5 35 M. 290 210 98 45 6th 40 M. 270 250 '95 70 7th 61 W. 240 200 95 45 8th 45 M. 210 190 100 80 15th 9 50 W. 300 240 116 60 10 64. W. 190 220 95 80 11th 64 W. 270.1 + 67.5 215 • 75 50 12th Average M. 190 ■ 130 59.16 + 16 M. 220 55 40.5% 20th 180 99.5 217.9 + 24.4 19.3%

Administration of 5 cm / day EPA alone results in
a decrease in
Serum levels of cholesterol and triglycerides, namely an average reduction of 11.2 for total cholesterol and of 15 percent for triglycerides. If pure cis-linoleic acid is administered for 45 days, the effect is
practically insignificant. In fact, many patients have cholesterol levels rising.

3 λ
However, when 3 cnr EPA + 2 cm cis-linoleic acid are administered per day, a pronounced synergistic effect can be determined. Using this combination results in a significantly significant reduction in serum cholesterol

-ΙΟΙ sterins (average decrease of 19.3 percent) and the Serum triglycerides (average decrease of 40.5 percent)

* 20

Claims (1)

CENTURY LABORATORIES, INC. Combined fatty acid composition to lower blood cholesterol and triglyceride levels ¹ ^ patent claims 1. Pharmaceutical composition for lowering cholesterol and triglyceride blood levels, consisting of . essentially of an effective amount of a combination of a first component from the group 5,8,11,14,17-eicosapentaenoic acid, 4,7,10,13,16,19-docosahexaenoic acid and combinations thereof and a second component from the group dihomo -jA-iinolenic acid, cis-linoleic acid, f- linolenic acid and combinations thereof, the first ² ^ and the second component in relative amounts of 3: 1 to 1: 3. are present. 2. Composition according to claim 1, characterized in that it additionally contains a pharmaceutically acceptable carrier. . 3. Composition according to claim 1, characterized in that it is essentially free of other unsaturated fatty acids.
. 4. Composition according to claim 1, characterized in that the first component is 5,8,11,14,17-eicosapentaenoic acid acts. 5. Composition according to claim 1, characterized in that that the second component is cis-linoleic acid. 6. Composition according to claim 4, characterized in that the second component is cis-linoleic acid acts. 7. Food containing a substantial amount of at least one fatty acid, characterized in that it the fatty acid is essentially a combination of a first component from the group 5,8,11, -14,17-eicosapentaenoic acid ', 4,7,10,13,16,19-docosahexaenoic acid and combinations thereof and a second component from the group consisting of dihomo- / '- linolenic acid, cis-linoleic acid, ^ -Linolenic acid and combinations thereof, the first and second components in relative amounts from 3: 1 to 1: 3 are available. 8. Food according to claim 7, characterized in that it is essentially free of other unsaturated substances Fatty acids is. 9. Food according to claim 7, characterized in that the first component is 5,8,11,14,17-Ej ^ cosapentaenoic acid acts. 10. Food according to .Anspruch 7, characterized in that that the second component is qis-linoleic acid acts. 11. Food according to claim 9, characterized in that the second component is cis-linoleic acid acts. 12. Composition according to claim 1, characterized in that it is the unsaturated contained therein Fatty acids are products made from natural sources len has been extracted by the following process steps are: - Iodination of the double bonds of the unsaturated fatty acids and triglycerides in the starting material; - saponification of the mixture obtained; - Extracting the iodized fatty acids from the saponified mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and - De-iodine the desired fraction. 13. Food according to claim 7, characterized in that it is the unsaturated contained therein Fatty acids are products that are extracted from natural sources through the following process steps have been: - Iodination of the double bonds of the unsaturated fatty acids and triglycerides in the starting material; - saponification of the mixture obtained; - Extracting the iodized fatty acids from the saponified mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and r De-iodine the desired fraction. 1 * t. Method of extracting pure fatty acids from natural ones Sources, characterized by the following process steps: - Iodination of the double bonds of the unsaturated fatty acids and triglycerides in the starting material .; - saponification of the mixture obtained; - Extracting the iodized fatty acids from the saponified mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and - De-iodine the desired fraction. 15. Composition according to claim 1, characterized in that it is the unsaturated contained therein Fatty acids are products that have been separated from fatty acid mixtures by the following stages: - Iodination of the double bonds of the unsaturated fatty acids in the mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and - De-iodine the desired fractions. 16. Food according to claim 7, characterized in that it is the unsaturated fatty acids contained therein Products that have been separated from mixtures of fatty acids by the following process steps: - Iodination of the double bonds of unsaturated fatty acids in the mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and ■ . - De-iodine the desired fractions. 17. A method for separating fatty acids from fatty acid mixtures, characterized by the following procedural steps: ''" _s - Iodination of the double bonds of the unsaturated fatty acids in the mixture; - methylation of iodinated fatty acids; - Separation of the fatty acids by column chromatography; and - De-iodine the desired fractions.