DE19700796C2 - Mixtures of active ingredients to reduce the cholesterol content in the serum of warm-blooded animals - Google Patents

Description

Field of the Invention

The invention relates to the use of mixtures of phytosterols and phytostanols or Phytosterol carboxylic acid esters and phytostanol carboxylic acid esters and selected potentiation means for reducing the cholesterol content in the serum of warm-blooded animals.

State of the art

Hypocholesterolemic agents are understood to mean agents that reduce the Cholesterol levels in the serum of warm-blooded animals result in no inhibition or Reduction in the formation of cholesterol in the blood occurs. For this purpose have already been developed by Peterson et al. in J. Nutrit. 50, 191 (1953) phytosterols, i.e. plant sterols, and their esters with Fatty acids suggested. The patent specifications US 3,085,939, US 3,203,862 and the German patent application DE-OS 20 35 069 (Procter & Gamble). The Active ingredients are usually added to frying or cooking oils and then through food recorded, but the amounts used are generally small and usually less than 0.5 % By weight in order to prevent the edible oils from clouding or the sterols from adding water be canceled. For use in the food sector, in cosmetics, pharmaceutical Preparations and in the agricultural sector are described in European patent application EP 0289636 A1 (Ashai) proposed storage-stable emulsions of sterol esters in sugar or polyglycerol esters. In The international patent application WO 92/19640 A1 (Raision) describes the incorporation of hydrogenated Phytosterol esters, namely sitostanol esters, to reduce blood cholesterol in Margarine, butter, mayonnaise, salad dressings and the like are proposed. Furthermore, be on the Writings Chem. Abstr. 79: 77319, DE 23 24 652 A1 and WO 96/38047 A1, which the Describe the use of phytosterols and tocopherols.  

The disadvantage, however, is that the phytosterol and phytostanol esters are the food can usually only be added in small quantities, otherwise there is a risk of that they affect the taste and / or the consistency of the agents. For sustainable However, influencing the cholesterol content in the blood would be the absorption of larger amounts Phytosterols and stanols or phytosterol and phytostanol esters are desirable. Farther The speed with which the substances maintain the level of cholesterol in the serum needs improvement Reduce. The object of the invention was therefore to remedy these shortcomings.

Description of the invention

The invention relates to the use of mixtures of active ingredients

• 1. Phytosterols and / or phytostanols and / or phytosterol carboxylic acid esters and / or Phytostanol carboxylic acid esters and
• 2. Chitosans and / or phytosterol sulfates and / or phytostanol sulfates and / or (deoxy) ribonucleic acids
  or
• 3. Phytostanols and / or phytostanol carboxylic acid esters and
• 4. Tocopherols and / or chitosans and / or phytosterol sulfates and / or phytostanol sulfates and (deoxy) ribonucleic acids

to reduce cholesterol levels in warm-blooded animals.

Surprisingly, it was found that tocopherols, chitosans, phytosterol sulfates, Phytostanol sulfates and / or deoxy or ribonucleic acids that do not have any have hypocholesterolemic properties, as potentiating agents for phytosterols and / or stanols and / or phytosterol and / or phytostanol carboxylic acid esters act, d. H. the decrease of the Cholesterol content in the serum in the presence of phytosterols and / or stanols and / or phytosterol and / or accelerate phytostanol carboxylic acid esters. Both encapsulated in gelatin Phytosterols and / or stanols and / or phytosterol and / or phytostanol carboxylic acid esters as also take the active ingredient mixtures orally without any problems.

Phytosterols, phytostanols, phytosterol carboxylic acid esters and phytostanol carboxylic acid esters

Phytosterols and phytostanols are to be understood as plant steroids that are only found on the C-3 Hydroxyl group, but otherwise no functional groups. They usually have them  Compounds 27 to 30 carbon atoms. Phytosterols are characterized by one Double bond in the ring system in 5/6, possibly 7/8, 8/9 or other positions. Phytostanols are the saturated compounds that can be obtained by hardening phytosterols. Typical examples of Suitable phytosterols are, for example, ergosterols, campesterols, stigmasterols, brassicasterols, preferably sitosterols or sitostanols and in particular β-sitosterols or β-sitostanols.

In addition to the phytosterols and phytostanols mentioned, their esters are preferably used. The acid component of the ester can be traced back to carboxylic acids of the formula (I)

R ¹ CO-OH (I)

in which R ¹ CO represents an aliphatic, linear or branched acyl radical having 2 to 22 carbon atoms and 0 and / or 1, 2 or 3 double bonds. Typical examples are acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, caprylic acid, 2-ethylhexanoic acid, capric acid, lauric acid, isotridecanoic acid, myristic acid, palmitic acid, palmoleic acid, stearic acid, isostearic acid, oleic acid, elaidic acid, petroselinic acid, linoleic acid, linoleic acid, linoleic acid, Ga doleic acid, behenic acid and erucic acid and their technical mixtures, the z. B. in the pressure splitting of natural fats and oils, in the reduction of aldehydes from Roe len's oxo synthesis or the dimerization of unsaturated fatty acids. Technical fatty acids with 12 to 18 carbon atoms such as coconut, palm, palm kernel or tallow fatty acid are preferred. The use of esters of β-sitosterol or β-sitanol with fatty acids having 12 to 18 carbon atoms is particularly preferred. These esters can either by direct esterification of the phytosterols with the fatty acids or by transesterification with fatty acid lower alkyl esters or triglycerides in the presence of suitable catalysts, such as. B. sodium ethylate or specifically also enzymes [cf. EP 0195311 A2 (Yoshikawa)].

Tocopherols

Tocopherols are taken to mean chroman-6-ols (3,4-dihydro-2-H-1-benzopyran-6-ols) which are substituted in the 2-position with 4,8,12-trimethyltridecyl radicals and follow the formula (II)

in which R ² , R ³ and R ^{4 are} independently hydrogen or a methyl group. Tocopherols belong to the bioquinones, i.e. polyprenylated 1,4-benzo and naphthoquinones, whose prenyl chains are more or less saturated. Typical examples of tocopherols which can be considered as component (b) in the context of the invention are ubiquinones, bovichinones, K vitamins and / or menaquinones (2-methyl-1,4-naphthoquinones). A distinction is also made in the tocopherols α, β, γ-, δ- and ε-tocopherols, the latter still having the original unsaturated prenyl side chain, and α-tocopherolquinone and hydroquinone, in which the pyran ring system is open. Preferably used as component (b) α-tocopherol (vitamin E) of the formula (II) in which R ² , R ³ and R ^{4 are} methyl groups, or esters of α-tocopherol with carboxylic acids having 2 to 22 carbon atoms, such as for example α-tocopherol acetate or α-tocopherol palmitate.

Chitosane

Chitosans are biopolymers and belong to the group of hydrocolloids. From a chemical point of view, these are partially deacetylated chitins of different molecular weights, which contain the following - idealized - monomer unit (III):

In contrast to most hydrocolloids, which are negatively charged in the range of biological pH values , chitosans are cationic biopolymers under these conditions. The positively charged Chitosans can and do interact with oppositely charged surfaces therefore in cosmetic hair and body care products as well as pharmaceutical preparations used (cf.Ullmann's Encyclopedia of Industrial Chemistry, 5th Ed., Vol. A6, Weinheim, Verlag Chemie, 1986, pp. 231-332). Overviews on this topic are also available from B. Gesslein et al. in HAPPI 27, 57 (1990), O. Skaugrud in Drug Cosm. Ind. 148, 24 (1991) and E. Onsoyen et al. in Seifen-Öle-Fette-Wwachs 117, 633 (1991). To make the Chitosane is assumed to be from chitin, preferably the shell remains of crustaceans, which are considered cheap Raw materials are available in large quantities. The chitin is produced in a process that for the first time by Hackmann et al. has been described, usually first by adding bases deproteinized, demineralized by adding mineral acids and finally by adding strong bases deacetylated, the molecular weights being distributed over a broad spectrum  can. Appropriate methods are, for example, made from Makromol. Chem. 177, 3589 (1976) or the French patent application FR A 2701266 known. In a preferred embodiment The invention is a chitin degradation product, as described in international patent application WO 96/16 991 A1 (Henkel) is described, or its degradation product is used with hydrogen peroxide. Out Chem. Abstr. 120: 392 the use of chitosans to lower cholesterol is known.

Phytosterol sulfates and phytostanol sulfates

Phytosterol sulfates and phytostanol sulfates are known substances, for example, by Sulphation of phytosterols or phytostanols with a complex of sulfur trioxide and pyridine can be produced in benzene [cf. J. Am. Chem. Soc. 63, 1259 (1941)]. Typical examples are the sulfates of ergosterols, campesterols, stigmasterols and sitosterols. The phytosterol sulfates and the phytostanol sulfates can be used as alkali and / or alkaline earth metal salts, as ammonium, Alkylammonium, alkanolammonium and / or glucammonium salts are present. As a rule, they will used in the form of their sodium salts.

(Deoxy) ribonucleic acids

Under (deoxy) ribonucleic acids (DNA or RNA) high molecular, thread-like Polynuc understood leotides derived from 2'-deoxy-β-D-ribonucleosides or D-ribonucleosides which in turn from equivalent amounts of a nucleobase and the pentose 2-deoxy-D-ribo furanose or D-ribofuranose can be built. The use of nucleic acids as active ingredients in the Cosmetics are well known. For example, in French patent application FR 2511253 A1 Skin and sunscreen containing high polymerized DNA suggested. From Japanese laid-open publication JP 62/096404 A2 (Kanebo) are cosmetic Compositions with nucleic acids and diisopropylamine dichloroacetate are known. Subject of French patent specification FR 2620024 B1 (Soc. d'Etudes Dermatologiques) are preparations which containing nucleic acid derivatives as radical scavengers. Examples are adenine, guanosine, xanthine, Hypoxanthine, uracil and ribonucleic acid. In international patent application WO 94/14419 A1 (Applied Genetics) skin and sun protection agents based on liposomes are proposed, which as special active ingredient contain a DNA repair enzyme. In the international patent application WO 95/01773 A1 (Boston University) uses a method to stimulate pigment production in which DNA fragments in liposomal form are transported into the epidermis. Finally, the subject of German patent application DE 43 23 615 A1 is compositions Containing nucleic acids and their fragments as anti-aging and sun protection creams.  

The DNA or RNA may contain the purine derivatives adenine and guanine and the pyrimidines cytosine and thymine or uracil as nucleobases. In the nucleic acids, the nucleobases are N-glycosidic with carbon atom 1 of the ribose, which in individual cases produces adenosines, guanosines, cytidines and thimidines. In the acids, a phosphate group links the 5'-hydroxy group of the nucleosides with the 3'-OH group of the following groups through a phosphodiester bridge to form single-stranded DNA or RNA. Due to the large ratio of length to diameter, DNA or RNA molecules tend to break strands even when subjected to mechanical stress, for example during extraction. For this reason, the molecular weight of the nucleic acids can range from 10 ³ to 10 ⁹ . For the purposes of the invention, concentrated DNA or RNA solutions are used which are distinguished by a liquid crystalline behavior. Deoxy- or ribonucleic acids are preferably used, which are obtained from marine sources, for example by extraction of fish sperm, and which have a molecular weight in the range from 40,000 to 1,000,000.

Industrial applicability

The active ingredient mixtures of the invention can be the phytosterols and / or stanols and / or Phytosterol and / or phytostanol carboxylic acid esters and the potentiating agents in a weight ratio 70: 25 to 25: 75 and particularly preferably contain 60: 40 to 40: 60, ensuring alone is that with the use according to the invention one for lowering the cholesterol content in the blood sufficient amount of component (a1) or (a2) is added. In a special one Embodiment of the invention, the active ingredient mixtures in a manner known per se in Encapsulated gelatin, with components (a1) and (b1) or (a2) and (b2) each in quantities from 0.1 to 50, preferably 1 to 30, in particular 5 to 25 and particularly preferably 10 to 15 parts by weight % - based on the weight of the gelatin capsules. A preferred embodiment for the oral ingestion of the active ingredients is the encapsulation of phytosterols and stanols and / or Phytosterol and phytostanol carboxylic acid esters in gelatin.

Another form of administration of the active ingredient mixtures are suppositories, which are rectal or vaginal can be performed, and as a supositorial matrix also gelatin, optionally in a combination nation with glycerin, or synthetic fats or waxes, polyethylene glycols or natural Components such as B. may contain cocoa butter.

In addition, it is possible to dissolve or disperse the mixtures in conventional foods as there are for example: salad oils, dressings, mayonnaises, margarines, butter, frying fats, cocoa pro products, sausage and the like.

Examples 1 to 16, comparative examples V1 to V4

There were gelatin capsules (weight approx. 1.5 g) with a content of 5 or 10% by weight of β-sitostanol or β-sitostanol esters and optionally 5 or 10% by weight of the various potentiating agents and 0.5% by weight of radiolabelled cholesterol. To study the hypocholeste The rinemic effect was allowed to fast male rats (individual weight approx. 200 g) overnight. At the the following day, the test animals were each crushed gelatin capsule with a little table salt containing water through a gastric tube. After 3, 6, 12, 24 and 48 h the animals became blood removed and the content of radioactive cholesterol determined. The results, the mean which represent measurements from 10 test animals are summarized in Tables 1 and 2. The Information on the decrease in radioactivity is in each case related to a blind group of Experimental animals, which only contain gelatin capsules containing 20% by weight of vitamin E and one appropriate amount of radiolabelled cholesterol had been administered. Mixtures 1 to 16 are according to the invention, the mixtures V1 to V4 are used for comparison.

Table 1

Hypocholesterolemic effect (quantities as% by weight based on gelatin capsule)

Table 2

Hypocholesterolemic effect (quantities as% by weight based on gelatin capsule)

Comparative examples V1 to V4 show that the addition of phytostanols or Phytostanol carboxylic acid esters the content of cholesterol in the blood of the test animals over a period of 48 hours significantly decreased. Components (b1) and (b2) accelerate according to Examples 1 to 16 the decrease in cholesterol and act as a potentiating agent for the phytostanols or phytostanol carboxylic acid esters.

Claims (10)

1. Use of active ingredient mixtures

• 1. Phytosterols and / or phytostanols and / or phytosterol carboxylic acid esters and / or phytostanol carboxylic acid esters and
• 2. Chitosans and / or phytosterol sulfates and / or phytostanol sulfates and / or (deoxy) ribonucleic acids
  or
• 3. Phytostanols and / or phytostanol carboxylic acid esters and
• 4. Tocopherols and / or chitosans and / or phytosterol sulfates and / or phytostanol sulfates and / or (deoxy) ribonucleic acids

to reduce the cholesterol level in the serum of warm-blooded animals. 2. Use according to claim 1, characterized in that as the phytosterol β-sitosterol, as phytosterol carboxylic acid ester β-sitosterol carboxylic acid ester, as phytostanol β-sitostanol and as Phytostanol carboxylic acid ester uses β-sitostanol carboxylic acid ester. 3. Use according to claims 1 and 2, characterized in that esters of β-sitosterol or β-sitostanol with carboxylic acids of the formula (I) are used,
R ¹ CO-OH (I)
in which R ¹ CO represents an aliphatic, linear or branched acyl radical having 2 to 22 carbon atoms and 0 and / or 1, 2 or 3 double bonds. 4. Use according to claim 3, characterized in that the carboxylic acids with fatty acids Are 12 to 18 carbon atoms.   5. Use according to claims 1 to 4, characterized in that tocopherols of the formula (II) are used as component (b2),
in which R ² , R ³ and R ⁴ independently of one another represent hydrogen or methyl groups. 6. Use according to claims 1 to 5, characterized in that one as a component (b2) uses α-tocopherol (vitamin E). 7. Use according to claims 1 to 6, characterized in that one as a component (b1 and b2) uses marine deoxyribonucleic acids which have a molecular weight in the range of Have 40,000 to 1,000,000. 8. Use according to claims 1 to 7, characterized in that the Components (a1) and (b1) or (a2) and (b2) in a weight ratio of 25:75 to 75:25 are used. 9. Use according to claims 1 to 8, characterized in that the Components (a1) and (b1) or (a2) and (b2) used encapsulated in gelatin. 10. Use according to claims 1 to 10, characterized in that the components (a1) and (b1) or (a2) and (b2) each in amounts of 0.1 to 50 wt .-% - based on the Weight of the gelatin capsules - inserts.