WO2025206981A1 - Composition for preventing iodine deficiency and optimizing iodine metabolism - Google Patents

Abstract

The invention relates to the medical and food industries, and more particularly to a composition for preventing iodine deficiency and optimizing iodine metabolism. The claimed composition for preventing iodine deficiency and optimizing iodine metabolism comprises, as an active agent, iodized micellar casein in an amount equivalent to 1-50 µg of iodine per kilogram of body weight per serving, in combination with a pharmaceutically acceptable carrier. The invention makes it possible to produce an effective composition that is highly soluble and exhibits long-term storage stability.

Description

NAME OF INVENTION

Composition for the prevention of iodine deficiency and optimization of iodine metabolism

AREA OF TECHNOLOGY

The invention relates to the medical and food industries, namely to a composition for the prevention of iodine deficiency and optimization of iodine metabolism.

PRIOR ART

A known method for producing a biologically active food supplement involves iodizing the original protein raw material by mixing it with an aqueous solution of molecular iodine at a ratio of inorganic iodine solution to the total protein content in the range of (2-40): 1. The iodination of the whey protein mixture is carried out with a mixture of enzymes immobilized on semipermeable membranes or on inert carriers, and the fermentation process is carried out with continuous monitoring of the iodine content in the solution. The solution of iodinated proteins is purified from impurities using macrofiltration, microfiltration, and ultrafiltration, followed by diafiltration of the aqueous solution of iodinated proteins in an ultrafiltration unit. The purified solution of iodinated proteins is subjected to sterilizing microfiltration, then freeze-drying to obtain the finished powder product. (RU 2212155 C1).

A method for producing iodinated whey proteins is known, which yields a biologically active substance by iodizing the starting protein raw material, which is α-lactalbumin, β-lactoglobulin, a mixture of these proteins, or hydrolysates of these proteins. Fermentation of the protein mixture with an aqueous solution of inorganic iodine is initiated by introducing a buffer mixture of reagents and an enzyme mixture based on lactoperoxidase, containing 16 to 24 wt.%, horseradish peroxidase, and 14 to 21 wt.% catalase. The fermentation process is carried out with continuous monitoring of the iodine content in the solution. The aqueous solution of iodinated proteins is purified from impurities by a combination of macrofiltration, microfiltration, and subsequent diafiltration of the aqueous solution of iodinated proteins in an ultrafiltration unit. The resulting solution of iodinated proteins is spray dried to obtain a finished powder product containing 0.5-4% deterministic covalently bound iodine in the form of a mixture contained in iodinated proteins. iodinated amino acids - monoiodotyrosines in the amount of 55-75 May. %, 24.0-43.5 May. % diiodotyrosines and 1.0-1.5 May. % triiodotyrosines (RU 2700444 C1).

However, the known methods have drawbacks in the form of a structurally complex process of enzymatic synthesis using immobilized enzymes, which leads to a high cost of the final product when using relatively inexpensive raw materials.

A harsh method of iodination of casein with iodine monochloride is known (OOO NIK MEDBIOFARM, TU 9229-001-48363077-99), which results in the production of a drug called “Iodcasein”.

Known methods produce iodinated proteins "Iodcasein" and "Bioiodine", a significant drawback of which is the following: more than 4/5 of the total amount of iodine present in the preparation "Iodcasein" is not represented by iodine-derivative amino acids, but by inorganic iodine compounds and esters of oxygen-containing acids - derivatives of positively charged iodine. Milk protein, heavily oxidized during hard iodination, is broken down by pancreatic intestinal proteases noticeably worse than the original casein, and only about 1/15 of the original amount of iodine in this preparation is released during proteolysis in the form of iodinated di- and tripeptides (with a m.v. of 0-500 Da), suitable for absorption by enterocytes - intestinal epithelial cells ("Conducting qualitative and quantitative analyses of samples of iodinated proteins "Bioiod" produced by Tekhnovita LLC and "Iodcasein" produced by Med biofar m LLC". RESEARCH REPORT. NON-STATE SCIENTIFIC CENTER High Technology Center "ChemRar", M., 2011. https://refdb.ru/look/2793682-pall.html).

A serious drawback of these iodized proteins is their complete insolubility (or extremely low solubility) in water: this feature greatly complicates the technological use of iodized proteins for the purpose of enriching food products with this microelement.

A method for producing an iodized food product is known, which uses diiodotyrosine in a free or bound state within a protein molecule for the prevention of iodine deficiency in the diet of food products fortified with it (RU 2134520 C1). The invention considers diiodotyrosine or its bound form in natural proteins as a chemically stable compound replacing unstable inorganic iodide compounds.

A "Biologically active food supplement for the prevention of iodine deficiency and optimization of iodine metabolism, and a food product containing it" (RU 2192150 C1) is known. The advantage of the invention lies in the proposal to use chemical compounds of various classes containing iodine in a covalently bound form as iodine-containing nutrients, including compounds iodinated at the 3- or 5-positions of the phenolic ring, such as monoiodotyrosine, diiotyrosine in the free state, or as part of protein and peptide molecules. The main feature of these compounds is their accessibility to enzymatic deiodination with the release of iodide, intended to meet the thyroid gland's needs for thyroid hormone synthesis. As iodine-containing nutrients, these compounds have an undeniable advantage over chemically unstable inorganic iodide salts.

The “Method for preventing damage by iodine radionuclides and optimizing iodine metabolism in the post-prophylactic period” (RU 2323733 C2) and “Method and preparation for preventing damage by iodine radionuclides to the human or animal body” (RU 2796757 C1) are known, which propose the use of proteins of plant or animal origin containing 4-hydroxy, 3-, 5-diiodophenyl compound as prophylactic preparations twice a day with a 10-14 hour interval.

However, the known methods do not envisage a specific product and only provide a list of possible raw materials for its creation. Another significant drawback is the proposal to use proteins containing covalently bound iodine at the 5- and 3- or 3-position of the phenol ring. It is known that a wide range of high-molecular compounds, especially when combined into "mixtures" (as indicated in the aforementioned patents), are heterogeneous materials that are extremely difficult to standardize for the subsequent production of a standardized product capable of serving as an effective means of preventing radiation injury.

Caseins, acid and rennet, are dry concentrates of food or technical quality, produced in the form of granulated or finely dispersed powders. Their isolation is based on the disruption of the colloidal stability of casein micelles, caused by the presence of a "hairy" layer of hydrophilic macropeptide residues of κ-casein on the micelle surface, which prevents their spherical approximation. During the precipitation process, in addition to monomeric denatured molecules, protein molecule complexes are formed, including dimeric, trimeric, and more complex forms, ultimately resulting in the presence of particles of varying sizes in the final product. The technologies used to produce such traditional protein ingredients (caseins, caseinates, co-precipitates) do not ensure the preservation of their native properties, the most important of which is solubility, which ensures high bioavailability and digestibility in the human gastrointestinal tract.

In the production of micellar caseins, ultrafiltration membranes are used, the pore size of which ensures the retention of almost all milk proteins, with the maximum removal of non-protein components. By using membranes of different sizes, it is possible to obtain micellar caseins with specified physicochemical properties, such as solubility, fat and moisture retention, emulsification, and also perform a number of technological functions in food systems (Kumar P, Sharma N, Ranjan R, Kumar S, Bhat ZF, Jeong DK. Perspective of membrane technology in dairy industry: A review. Asian-Australasian Journal of Animal Science. 2013;26(9): 1347-1358. https://doi.org/10.5713/ajas.2013.13082).

DISCLOSURE OF THE INVENTION

The objective of the invention is to create a composition for the prevention of iodine deficiency and optimization of iodine metabolism based on iodinated micellar casein.

The technical result of using the proposed invention consists in obtaining iodinated micellar casein and creating an effective composition based on it for the prevention of iodine deficiency and optimization of iodine metabolism, which has a therapeutic and prophylactic effect and an unchanged composition during long-term storage.

The stated problem was solved by creating a method for obtaining iodinated micellar casein, suitable for use in the compositions of pharmacological and veterinary drugs, biologically active additives and radioprotective drugs, including the addition of crystalline potassium iodide to an aqueous solution of micellar casein in a ratio of 1:7 - 1:9, holding the mixture for 5-10 minutes, adding crystalline potassium persulfate to the mixture in a ratio to the weight of micellar casein of 1:8 - 1:12, then holding the mixture for 4-8 hours at a temperature of 18-25 °C and a pH of 6.5-8.9 with continuous monitoring of the content of molecular iodine in the solution and periodically adding portions of crystalline potassium persulfate in a ratio to the weight of micellar casein of 1:12 - 1:24 to continue the iodination reaction, purifying the resulting aqueous solution of iodinated micellar casein from the remaining inorganic salts by microfiltration and diafiltration, followed by sterilizing microfiltration, sublimation or spray drying.

The stated problem was solved by creating a composition for the prevention of iodine deficiency and optimization of iodine metabolism, containing iodinated micellar casein as an active substance in an amount that contains, in terms of iodine, 1-50 μg per kg of body weight for a single dose, in combination with a pharmaceutically acceptable carrier.

It is preferable that the composition for the prevention of iodine deficiency and optimization of iodine metabolism is made in the form of capsules, tablets, powder, solutions, or is included in vitamin-mineral complexes.

The micellar casein used in the proposed composition surpasses rennet casein and all types of animal and plant protein mixtures—egg albumin, milk or whey proteins, hemoglobin, soy protein, yeast proteins, and yeast hydrolysate—in its physicochemical properties. This is because micellar casein molecules are uniform in size and have nearly 100% solubility.

Table 1. Some physicochemical properties of proteins, including the micellar casein fraction (according to our own research). This also explains the high efficiency of iodinated micellar casein, as micellar casein has a solubility close to 100% and retains the protein's native properties, enabling "soft iodination" at solution pH values that preserve the protein's native structure. Preservation of the protein's native structure is also ensured by the selected reagent concentration ranges, the fractional size, and the order of their addition, which avoids the effects of extreme concentrations of highly active substances on the protein.

Iodinated micellar casein used in the claimed composition is obtained as follows.

The required amount of crystalline potassium iodide is added to a 5-10% aqueous solution of micellar casein, mixed, and the mixture is left to stand for 5-10 minutes. Crystalline potassium persulfate is then added at a ratio of 1:8 to 1:12 to micellar casein, and the mixture is left to stand for another 4-8 hours. The molecular iodine content in the solution is continuously monitored, and portions of crystalline potassium persulfate are periodically added to continue the iodination reaction. Iodination is carried out at a temperature of 18-25°C, in a pH range of 6.5-8.9, with constant pH monitoring and titration maintaining the required value.

An aqueous solution of micellar casein with covalently bound iodine is purified from remaining inorganic salts using microfiltration followed by diafiltration of the aqueous solution. The resulting iodinated micellar casein solution undergoes sterilizing microfiltration, followed by freeze-drying or spray drying to yield a finished powder product with a covalently bound iodine content of 4-10%.

The method yields a product containing 4-10% deterministic covalently bound iodine in the form of iodinated amino acid monomers. Iodine is covalently bound at the 5- and 3- or only at the 3-position of the phenolic group of tyrosinyl monomers, which is part of micellar casein, as well as at the 2-, 2-, and 5-positions, as well as the 1-, 2-, and 5-positions of the histidinyl monomers, which is part of micellar casein.

Experimental studies of iodinated micellar casein for use in the formulations of pharmacological and veterinary drugs, as well as Dietary supplements have shown their high effectiveness. This was confirmed by animal experiments.

Purified iodinated micellar casein is used to produce pharmaceutical and veterinary formulations for the prevention of iodine deficiency and the optimization of iodine metabolism. For the production of formulations, particularly dietary supplements (DS), for the prevention of iodine deficiency and the optimization of iodine metabolism, iodinated micellar casein is available in both monoforms and combinations with other physiologically active substances, providing a customized approach to the prevention of diseases associated with iodine metabolism disorders. These formulations are available in capsules, tablets, powders, solutions, or as part of vitamin and mineral complexes.

IMPLEMENTATION OPTIONS OF THE INVENTION

Method for obtaining the proposed composition.

Dried iodinated micellar casein, containing the daily human iodine requirement (150 mcg), is mixed with suitable carriers. The resulting mixture is either packaged as a powder, or tableted and packaged, or placed in capsules (such as hard gelatin capsules for medical use and packaged), or dissolved in water and bottled.

Below are examples of the production of the proposed composition in the form of vitamin-mineral complexes.

Example 1.

Dried iodized micellar casein in an amount that provides the daily requirement for iodine (150 mcg) is added to the bars, containing additional proteins, carbohydrates and fats as fillers, as well as premixes of vitamins and mineral components.

Example 2.

Dried iodized micellar casein in an amount that provides the daily requirement for iodine (150 mcg) is mixed with carriers, as well as premixes of vitamins and mineral components, and packaged in the form of capsules and tablets. Example 3.

Dried iodized micellar casein is mixed with premixes of vitamins, minerals, and flavorings, using sucrose as a carrier. The resulting product is used as a powder for making a drink.

Below are examples of some dietary supplement compositions:

A monoform of the drug, produced in capsules.

Capsule contents weight: 100-500 mg

Ingredients: iodinated micellar casein 5% - 1-30 mg, microcrystalline cellulose - 90-492 mg, magnesium stearate - 1-5 mg.

Soluble form, for sublingual instillation:

Volume of solution in a bottle: 15-50 ml,

Ingredients: distilled water 12-45 ml, glycerin - 2-5 g, iodinated micellar casein 5% - 30-500 mg.

Tablet.

Tablet weight: 100-500 mg,

Ingredients: iodinated micellar casein 5% - 1-3 mg, microcrystalline cellulose - 90-492 mg, magnesium stearate - 1-5 mg.

Powder, in sticks, weight 9-18 g.

Ingredients: sucrose - 6-17.6 g, edible salt - 0.1-0.28 g, vitamin C - 15-43 mg, vitamin B3 - 3-8 mg, vitamin E - 2-6 mg, vitamin B5 - 1-3 mg, vitamin B6 - 0.01-0.61 mg, vitamin B2 - 0.01-0.61 mg, vitamin B 1 - 0.01-0.6 mg, vitamin B9 - 0.1 mg, vitamin B 12 - 0.001 mg, iodized micellar casein 5% - 0.5-1 mg, magnesium bisglycinate (1000 mg magnesium) - 0.5-1 g, flavoring - 0.1 -0.2 g.

The claimed composition obtained on the basis of iodinated micellar casein has good water solubility and stability in iodine content during long-term storage.

Table 2. Iodine content in iodinated micellar casein (IMC) during storage.

To demonstrate the efficacy of the iodinated micellar casein supplement, biological experiments were conducted in accordance with the recommendations of MUK 2.3.2.721 "Determination of the safety and efficacy of dietary supplements." The experiment was conducted on male Wistar rats weighing 200 g, with n = 8-10 rats in each group.

Considering the pathogenetic mechanisms of hypothyroidism development, a hypothyroidism model was created to experimentally simulate its main symptoms. For this, mercazolil, which accelerates the removal of iodides from the thyroid gland, inhibits peroxidase, and inhibits the iodination of tyrosine to form thyroid hormones, was administered at a dose of 25 mg/kg of animal body weight.

In the experiment, the concentration of thyroid hormones in the blood serum of experimental animals was determined against the background of hypothyroidism and with subsequent correction with a preparation of iodinated micellar casein in the form of a composition (example 1) and The reference drug was potassium iodide. The iodine concentration was calculated based on the animal's weight and corresponded to the WHO-recommended human dose of 200 μg iodine. Hormone levels were determined using enzyme-linked immunosorbent assay.

Chronic administration of mercazolil to rats for 2 weeks caused a 2-fold decrease in the concentration of thyroxine and triiodothyronine, while the level of thyroid-stimulating hormone increased 2-fold (Table 3).

Table 3. Indicators of decrease in thyroxine and triiodothyronine concentrations.

Note: p < 0.01 (mean error).

When exposed to a composition with iodinated micellar casein, the level of hormones: thyroxine was 99.8%, triiodothyronine - 95.5%, thyroid-stimulating hormone - 95.2% of the level of hormone concentration in the intact group of animals.

The use of KI-200 for the correction of experimental hypothyroidism reduced the concentration of thyroid-stimulating hormone by 35% and increased the concentration of thyroxine by 55% and triiodothyronine by 45% compared to the experimental hypothyroidism group.

Thus, experimental data show that the claimed composition containing iodinated micellar casein completely eliminates the effects of hypothyroidism and is much more effective than KI.

Claims

Invention formula 1. A composition for the prevention of iodine deficiency and optimization of iodine metabolism, containing iodinated micellar casein as an active substance in an amount that contains, in terms of iodine, 1-50 μg per kg of body weight for a single dose, in combination with a pharmaceutically acceptable carrier. 2. The composition according to claim 1, characterized in that it is made in the form of capsules, tablets, powder, solutions, or is part of vitamin-mineral complexes.