RU2765999C1 - Biologically active food additive containing non-ionic forms of iodine and selenium - Google Patents

Abstract

FIELD: food industry; pharmaceutics.

SUBSTANCE: invention refers to food industry, medical and veterinary pharmacology. Preparations obtained by the proposed method can be used as food additives in the production of bakery and confectionary products, dairy and fat-and-oil products, fruit water and other beverages, canned food, vitamin and microelement complexes, feedstuffs and premixes for animals. Biologically active additive is an aqueous solution of non-ionic compounds of iodine and selenium, wherein iodine is contained in it in form of 3,5-diiodo-L-tyrosine, and selenium atoms are covalently bonded in an inorganic lattice of iron-dextrin nanocolloidal particles with the following content of iodine and selenium in grams per 1 l of the solution: iodine – 1–50, selenium – 1–10.

EFFECT: biologically active additive has high chemical stability and composition invariance during long-term storage or culinary treatment, independence of the degree of absorption of the microelement from the characteristics of the main diet of the consumer, absence of undesirable chemical interactions of microelement additives with each other or with other mineral or organic components of food during its assimilation in the gastrointestinal tract.

1 cl, 1 tbl

Description

The invention relates to the field of food industry, medical and veterinary pharmacology. The resulting products can be used as food additives in the production of bakery and confectionery products, dairy and fat-and-oil products, fruit waters and other drinks, canned food, vitamin and microelement complexes, animal feed and premixes.

The problem of alimentary deficiency of iodine and selenium is currently considered extremely acute and relevant. In animals and humans, iodine is part of the so-called thyroid hormones (thyroxine and triiodothyronine), produced by the thyroid gland and having a multifaceted effect on the growth, development and metabolism of the body.

Selenium is a synergist of vitamin E and iodine: with selenium deficiency, iodine is poorly absorbed by the body, and its biotransformation into triiodothyronine is greatly slowed down (which is due, in particular, to a lack of concentration of 5'-iodothyronine deiodinase, a key selenium-containing enzyme in thyroid tissues that converts thyroxine into significantly more active triiodothyronine) (The interactions between selenium and iodine deficiencies in man and animals. Arthur JR, Beckett GJ, Mitchell JH. - Nutrition Research Reviews. 1999 Jun; 12(1):55-73).

In the vast majority of cases (Federal Register of Biologically Active Food Supplements. Section 3: "Biologically Active Supplements (BAA) to Food - Sources of Mineral Substances". Moscow, 2001; Rebrov V.G., Gromova O.A. Vitamins, macro- and microelements: training programs of the RSC of the UNESCO Institute of Microelements Publisher: GEOTAR-Media, 2008 C.587-596 https://www.elibrary.ru/item.asp?id=19497570) currently registered dietary supplements (as well as similar medicinal preparations and premixes for animals not discussed in these references) are mixtures of water-soluble inorganic compounds of iodine (iodide or, more rarely, potassium iodate) and selenium (selenate or sodium selenate). The exceptional popularity of such combinations is due to the simplicity and low cost of preparing such compositions, but their serious shortcomings are also well known. There is a relatively high toxicity of mineral preparations of iodine and selenium (LD ₅₀ for selenium compounds - 4-5 mg / kg, for iodine compounds - 60-100 mg / kg) and, most importantly, chemical instability and a tendency to secondary pharmacokinetic and pharmacodynamic interactions with inorganic components of the atmosphere, water, food or semi-finished food.

Indeed, the oxidation of the iodide ion with atmospheric oxygen, followed by the loss of volatile molecular iodine, is the reason for the short shelf life of iodine-containing dietary supplements, including the most famous of them, iodized salt. The shelf life of this additive is 6 months, however, depending on the storage conditions, a twofold decrease in the concentration of iodide ions in this product can be observed after 3-10 months; when salt is iodized with potassium iodate, the shelf life of table salt increases slightly. Even faster is the process of iodine loss from polymicroelement dietary supplements, drugs or premixes for animals containing, along with iodides and iodates, appreciable amounts of transition metal ions (compounds of iron, manganese, cobalt, copper, etc. - catalysts for redox reactions involving iodine-containing ions).

Inorganic iodine compounds, as a rule, do not withstand any severe conditions of culinary food processing: even short-term boiling of their aqueous solutions leads to an irreversible loss of 30-60% of the initial amount of iodine. In addition, mineral iodine is highly toxic: the maximum allowable daily intake of iodine in the Russian Federation is 300 mcg (MR No. 2.3.1.1915-04 of the Ministry of Health and Social Development of the Russian Federation, Moscow, 2004, 34 s), the WHO recommendation is no more than 1000 mcg per day.

Iodine- and selenium-containing dietary supplements made on the basis of seaweeds and products of their processing are considered to be much safer and more effective. Indeed, such preparations have been used since ancient times in China, Japan and Southeast Asia to eliminate the painful manifestations of iodine deficiency, they are relatively cheap, effective and low-toxic, and a sufficiently high concentration of iodine in the dry mass of brown algae allows one to limit oneself to small daily doses of such substances. In addition, a significant part of iodine (15-25% of its total content) is present in algae tissues already in organic form - in the form of 3-iodine-L-tyrosine and 3.5-diiodine-L-tyrosine amino acid fragments of the corresponding plant proteins. The covalent bond "carbon-iodine" in such substances is very strong, and iodine from the composition of aromatic iodine-containing amino acids is no longer lost either during storage or during heat treatment, does not show undesirable chemical interactions during the storage of the drug or during the assimilation of the microelement from the gastrointestinal tract. tract.

Unfortunately, the total iodine content in brown algae (saccharine kelp, palmate or toothed kelp, vesicular fucus, pinnate undaria, etc.) varies within a fairly wide range - 0.005-0.25% of dry weight, depending on growing conditions. The predominant part of iodine (about 4/5 of the total amount) is still contained in algae in ionic form - in the form of iodide and iodate anions - and therefore can also be lost during the technological preparation of raw materials, their long-term storage or culinary processing. In addition, the intestinal digestive enzymes of mammals are not able to cleave the peptide bonds formed by mono- and diiodotyrosines (in contrast to the peptide bonds of the original tyrosine or other aromatic amino acids): brown algae proteins are much inferior to the proteins of land agricultural plants in terms of digestibility, and a significant part of the "organic iodine" of seaweed by man or higher animals cannot be absorbed.

A modern modification of this approach (that is, the use of stable organoiodine compounds as dietary supplements) is very interesting: chemical iodination of tyrosine residues of one of the milk proteins by the action of free iodine or iodine monochloride on a dilute aqueous protein solution. Thus, the drug called "Iodkazein" (LLC "NPK MEDBIOPHARM", TU 9229-001-48363077-99) - a product of hard iodization of casein with iodine monochloride - contains 7-10% iodine and is recommended, in particular, for use in food production for enrichment of various food products with "organic iodine". For individual use, the preparation "Bioiodine" (LLC "TECHNOVITA", TU 9141-001-69752570-11), obtained by mild enzymatic iodination of milk albumins (1.5-2.0% iodine), is proposed.

In fact, more than 4/5 of the total amount of iodine present in the most popular preparation "Iodcasein" (recommended by the Ministry of Health, the Russian Academy of Medical Sciences and the Ministry of Agriculture of the Russian Federation for use as a dietary supplement for both individual use and in the food industry: MP 2.3.7.1916-04 The use of iodcasein for the prevention of iodine deficiency diseases as a means of population, group and individual prevention of iodine deficiency.METHODOLOGICAL RECOMMENDATIONS of the Ministry of Health, 2004. amino acids, and inorganic compounds of iodine and esters of oxygen-containing acids - derivatives of positively charged iodine. Milk protein, highly oxidized during hard iodination, is cleaved by intestinal pancreatic proteases significantly worse than the original casein, and only about 1/15 of the initial amount of iodine in this preparation is released during proteolysis in the form of iodinated di- and tripeptides (with m.v. 0 -500 Da), suitable for assimilation by enterocytes - epithelial cells of the intestine ("Qualitative and quantitative analyzes of samples of iodinated proteins "Bioiodine" produced by Technovita LLC and Iodcasein produced by Medbiopharm LLC". REPORT ON SCIENTIFIC RESEARCH WORK. NON-STATE SCIENTIFIC CENTER Center for High Technologies "KhimRar", M., 2011. https://refdb.ru/look/2793682-pall.html).

A serious drawback of iodinated proteins is their complete insolubility (or extremely low solubility) in water: this feature greatly complicates the technological use of iodinated proteins in order to enrich food production with this microelement. Proteins of brown algae do not dissolve in water at all under any conditions.

This methodological approach (the introduction of a heteroelement into the amino acid sequence of a natural protein) has also been successfully used to create selenium-containing (and not just iodine-containing) dietary supplements. Indeed, the cultivation of some saccharomycete yeasts in the presence of an excess of sodium selenite leads to a partial incorporation of the organoelement amino acid residues of selenocysteine Se-CYS and selenomethionine Se-MET into the amino acid sequence of the corresponding yeast proteins (usually Se-CYS and Se-MET are included in the protein in a molar ratio of ~ 1:2). The dry biomass of such yeast, as well as preparations of partially purified or autolyzed yeast protein, are sometimes used as a source of dietary selenium (for example, imported preparations of dietary supplement “Iodoselen” (“Paragon Laboratories Inc.”, USA), SGR: No. 77.99.11.003.E. 009774.10.15 dated October 23, 2015; dietary supplement "SELENIUM" ("Archon Vitamin Corporation", USA), SGR No. 77.99.23.3.U.4667.4.05 dated April 28, 2005; dietary supplement "SELENIUM SOLGAR" ("Solgar Vitamin", USA) ), SGR No. 77.99.23.3.U.5465.6.09 dated 06/03/2009, etc.)

The toxicity parameters of "organic yeast selenium" do not differ too much from those of compounds of tetra- and hexavalent inorganic selenium (in terms of the content of pure selenium), and in terms of effective digestibility, inorganic selenium-containing dietary supplements even significantly outperform preparations of "organic selenium" - not to mention already about the disproportionately higher cost of the daily dose of the active substance in the latter case. Nevertheless, “organic selenium” preparations have, in comparison with water-soluble selenites and alkali metal selenates, one undoubted and extremely important advantage - TOXIC SAFETY: their use minimizes the risk of selenium intoxication in case of accidental overdose of the drug by an incompetent consumer.

The closest to the proposed drug (prototype) is the drug Iodoselenium containing vitamin C (potassium ascorbate or magnesium ascorbate), vitamin E (30% mixture of d-tocopherols: alpha, beta, gamma, sigma), iodine (from kelp extract), selenium (selenomethionine) (see "Iodoselen", web page https://vitamax-moscow.ru/jodoselen).

The disadvantages of this preparation is that it is difficult to use it to enrich other products, since this preparation cannot be dissolved in water. Also, the main part of the iodine from the kelp extract is contained in the form of iodide and iodate anions - and therefore can be lost during the technological preparation of raw materials, their long-term storage or culinary processing. In addition, the intestinal digestive enzymes of mammals (including humans) are not able to cleave the peptide bonds formed by mono- and diiodotyrosines, so that part of the "organic iodine" from seaweed (kelp) cannot be absorbed by humans or higher animals.

The technical problem solved by the invention is to create a microelement biologically active food supplement that has:

• high chemical resistance and invariability of the composition during long-term storage or cooking;

• independence of the degree of assimilation of the microelement from the characteristics of the main diet of the consumer, the absence of undesirable chemical interactions of microelement supplements with each other (or with other mineral or organic food components) in the process of its assimilation in the gastrointestinal tract;

The technical problem is solved by a biologically active food supplement, which is an aqueous solution containing iodine and selenium in the form of non-ionic compounds, while the supplement contains iodine in the form of 3,5-diiodine-L-tyrosine, and selenium - as part of an iron-dextrin complex, with the following content of iodine and selenium in grams per 1 liter of solution:

iodine 1 - 50

selenium 1 - 10

Replacement of iodinated proteins of milk or algae (that is, high-molecular compounds, including only individual rare fragments of mono-3-iodine-L-tyrosine or di-3,5-diiodine-L-tyrosine) with chemically individual monomeric forms of mono-3-iodine-L -tyrosine or di-3,5-diiodine-L-tyrosine (low-molecular compounds that do not require preliminary depolymerization of the drug by the action of digestive tract proteases) allows us to offer an iodine-containing dietary supplement that combines the following advantages: high heteroelement content, water solubility, toxic safety, chemical stability during storage and biological assimilation, devoid of the main disadvantages of the above-mentioned drugs.

A technique that is absolutely similar in biochemical terms (it can be called "diversification of the ways of delivering a trace element to an enterocyte") can also be applied to the case with a selenium-containing dietary supplement: here it is also proposed to use a derivative of covalently bound selenium, which is absorbed by the cells of the intestinal epithelium according to a completely different, non-ionic mechanism ( fundamentally different from the banal transmembrane transport of selenite or selenate ions). Indeed, nanoparticles of colloidal iron-dextrin preparations such as "MALTOFER", "FERRUM LEK", "BIOIRON" are assimilated by enterocytes due to the same mechanism as micelles of water-insoluble food lipids - by pinocytosis (capture of a drop of the liquid phase with particles of the solid phase contained in it and subsequent enzymatic cleavage of trapped particles in the intracellular space of the enterocyte).

Refusal to use the simplest ionic forms of the mentioned trace elements (I ^- , IO ₃ ^- , SeO ₃ ^2- , SeO ₄ ^2- ) made it possible to create a combined iodine-containing dietary supplement, characterized by:

• very low acute toxicity and guaranteed toxic safety;

• high chemical resistance and composition stability during long-term storage;

• independence of the degree of assimilation of the microelement from the characteristics of the main diet of the consumer, the absence of undesirable chemical interactions of microelement supplements with each other (or with other mineral or organic food components) in the process of its assimilation in the gastrointestinal tract;

• high concentration of target microelements in preparations;

• high solubility of drugs in water, which eliminates the need to perform any operations for the preliminary preparation of drugs for their use in food production.

Iodine and selenium are biochemical synergists, and the daily norms of their consumption by a person correlate approximately as 2:1 - 4:1 (120-150 micrograms of iodine versus 40-60 micrograms of selenium). Thus, the proposed drug can be considered a synergistic composition of a pair of trace elements, iodine and selenium, in their non-ionic forms. It is a homogeneous red-brown aqueous solution with pH = 8.0-10.0, containing a non-ionic form of iodine in the form of its organoelement compound 3,5-diiodine-L-tyrosine and a non-ionic form of selenium included in the Fe ₂ O covalent lattice ₃ iron-dextrin nanoparticles.

The drug eliminates the lack of iodine and selenium in the diets of people and animals, and its distinguishing feature is the complete absence of ionic forms of these microelements (iodide, iodate, periodate, selenide, selenite, selenate ions). Iodine is present in the proposed preparation in the form of a highly physiological amino acid 3,5-diiodine-L-tyrosine (that is, an intermediate product of the biochemical synthesis of thyroid hormones from iodide ions): the molecule of this compound includes a strong covalent bond between the iodine atom and the carbon of the aromatic cycle.

Covalently bound atoms of tetravalent selenium are introduced into the composition as part of an iron-dextrin nanocolloid (in this case, >Se=O structural fragments replace iron atoms in the Fe ₂ O ₃ oxide crystal lattice of the inorganic core of the nanoparticle) (see RU 2409375 C1, pub. 20.01.2011 ).

The proposed micronutrient dietary supplement is prepared as follows.

A concentrated dextrin solution containing Fe (III) chloride and sodium selenite (the composition and proportions for each group of animals or humans are established according to the established recommendations), neutralize with stirring with a concentrated soda solution until a precipitate begins to form, alkalinize the mixture with a solution of sodium hydroxide to pH 9-11 , the mixture is heated to the temperature of dissolution of the precipitate, the resulting red-brown solution is acidified with concentrated HCl to pH 5.8-6.5. The concentration and desalting of the colloidal solution is carried out by ultrafiltration on membranes with an exclusion limit of more than 10 kDa.

The preparation of stable nanocolloids of iron-dextrin modified with selenium (covalently included in the iron oxide crystal lattice at a concentration of 0.1-20.0 wt% of the iron content) is described in more detail in patent RU 2409375 C1. Spectrophotometric control of the composition of the resulting preparation indicates the quantitative covalent binding of selenite ions introduced into the reaction mixture by the crystal lattice of iron oxide (III) - the basis of the inorganic core of nanoparticles of iron-dextrin colloids.

EXAMPLE OF PREPARATION OF BAA CONTAINING IODINE AND SELENIUM IN CONCENTRATIONS OF 10 g/L AND 4 G/L RESPECTIVELY

INITIAL COMPONENT 1: SELENE-CONTAINING IRON-DESTRIN NANOCOLLOID. A concentrated nanosuspension of selenium-containing iron-dextrin nanoparticles is prepared in accordance with the method given in the patent "Method for obtaining a microelement drug based on an iron-dextrin complex", RU No. 2409375. At the same time, 50.0 kg of ferric chloride hexahydrate, 1.20 kg of sodium selenite and 30 kg of dextrin with a molecular weight of 2-5 KDa are used as starting materials. Receive 100 l of liquid concentrated nanocolloid containing 100 g/l of non-ionic iron and 6.1 g/l of covalently bound selenium.

STARTING COMPONENT 2: CRYSTALLINE 3,5-DIIODO-L-TYROSINE. An improved method for the mild iodination of L-tyrosine in acetic acid medium (see US patent 2,835,700, May 20, 1958) was used to synthesize the 3,5-diiodo-L-tyrosine preparation: a typical sample, judging by the results of chromatographic and volumetric analysis, contains 97-98 % of the target substance, 1-3% mono-3-iodine-L-tyrosine and trace, at the level of 0.2%, the concentration of the initial tyrosine, acetates and iodides of alkali metals (melting point of the anhydrous preparation - 230-234 ° C, with decomposition ).

INTERMEDIATE 3: WEAKLY BASIC SOLUTION OF 3,5-DIIODO-L-TYROSINE. To a mixture of 60 g of crystalline 3,5-diiodo-L-tyrosine and 0.5 l of water, a concentrated solution of sodium hydroxide was added with stirring until the amino acid precipitate was completely dissolved, the volume of the solution was adjusted to 1.00 l, and the pH value was up to 9.6 -9.8. The concentration of non-ionic iodine in the intermediate product is 30 g/L.

FINAL PRODUCT 4 - BAA was mixed with 330 ml of an alkaline solution of 3,5-diiodo-L-tyrosine (INTERMEDIATE PRODUCT 3) with a concentration of non-ionic iodine equal to 30 g/l, and 670 ml of a solution of selenium-containing iron-dextrin nanocolloid containing selenium at a concentration of 6 g/l (INITIAL COMPONENT 1). The resulting solution containing 4 g/l of selenium and 10 g/l of iodine was pasteurized and preserved by adding 1.5 g of methylparaben (sterilization at 110°C or 120°C is also acceptable).

EXAMPLES OF BAA COMPOSITIONS

The table shows examples of the proposed dietary supplement with a different content of trace elements.

table

trace element Microelement content, g/l dietary supplement 1 dietary supplement 2 dietary supplement 3 dietary supplement 4 Iodine one eleven thirty 50 Selenium one 4 7 10

The proposed dietary supplement of all the above compositions has good solubility, chemical and pharmacodynamic stability. Both the selenium-containing component of this synergistic composition (iron-dextrin colloid) and its iodine-containing component (3,5-diiodine-L-tyrosine) are highly soluble (up to 15-20%) and quite stable during storage. The characteristics of dietary supplements and its components do not change during storage for 3-4 years, according to the "accelerated aging" method (I-42-2-82 INTERIM INSTRUCTIONS FOR WORK TO DETERMINE THE EXPIRATION DATES OF MEDICINAL PRODUCTS ON THE BASIS OF THE "ACCELERATED AGING" METHOD AT HIGH TEMPERATURE INTRODUCED BY Order of the Ministry of Health of the USSR and the Ministry of Medical Industry on April 18, 1983 No. 430/224.) in weakly alkaline aqueous solutions at pH 8.0-9.5. The components of the drug do not show any tangible interactions with each other, as well as with the protein, lipid and carbohydrate components of food products during boiling in water.

The drug is completely indifferent in relation to the components of feed and premixes and can be used as a source of trace elements in feed production and in veterinary pharmacology.

The use of dietary supplements for the enrichment of food or animal feed (both in the course of their individual use and in the technological processes of food production) does not require any preliminary preparation. An appropriate volume of liquid formulation is simply mixed into the aqueous phase of the micronutrient-enriched food product - or the formulation is applied by spraying the surface of the solid particles of the dry food (feed) product.

The extremely low toxicity of the proposed composition makes it impossible to correctly determine the LD ₅₀ values: at no physically achievable doses of the finished drug - or each of its two individual components - the death of experimental animals was recorded (tests were carried out on white rats of both sexes weighing 200 ± 25 g at oral administration of the maximum possible doses of concentrated drugs through a gastric tube). It was only possible to state that the LD ₅₀ value for pure selenium (which is part of the selenized iron dextrin), in any case, significantly exceeds 150 mg / kg, and the LD ₅₀ value for pure iodine (in the composition of 3,5-diiodine-L- tyrosine component) exceeds 1000 mg/kg. Thus, the toxicity of the drug for selenium (in terms of a microelement mass unit) is at least 40-50 times less than the toxicity of inorganic selenium compounds, and its toxicity for iodine is at least 10-15 times less than that of inorganic compounds. iodine compounds.

EXAMPLE OF APPLICATION OF BAA. The proposed drug with the content of non-ionic forms of iodine and selenium, equal to 11 mg/ml and 4 mg/ml, respectively (composition No. 2 of the table), was used to enrich drinking milk with these microelements (at concentrations of 200 μg/l and 80 μg/l, respectively). For this purpose, 20 ml of the drug was added to 1000 liters of milk and mixed: 0.5 liters of such a product contains an approximately daily dose of each of the indicated microelements.

Used dietary supplements of other compositions indicated in the table. All of them allow you to create a daily dose of each of these trace elements with an appropriate ratio of the volume of the drug and milk.

The proposed solution to the problem of deficiency of these microelements (the use of their water-soluble non-ionic forms) provides the possibility of reliable targeted delivery of iodine and selenium to the human or animal body, makes it possible to achieve the chemical stability of its biologically active components during storage, cooking and digestive assimilation of preparations, eliminates the dependence of the degrees of assimilation trace elements from the characteristics of the main diet of the consumer.

The preparative form of this composition is ready for direct use (both for individual use and in technological processes of food or feed production) and does not require any preparatory operations.

Claims (2)

Biologically active food supplement, which is an aqueous solution containing iodine and selenium in the form of non-ionic compounds, characterized in that it contains iodine in the form of 3,5-diiodine-L-tyrosine, and selenium - as part of the iron-dextrin complex with the following content iodine and selenium in grams per 1 liter of solution: iodine 1-50 selenium 1-10