RU2457852C2 - New application of therapeutically effective peptides - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: tripeptides Ile-Pro-Pro (IPP) and/or Val-Pro-Pro (VPP) are applicable for preparing a product removing arterial stiffness and for preparing a product improving vascular elasticity. They are made of casein or casein-containing starting material by fermentation with the strain Lactobacillus helveticus DSM 13137 or Lactobacillus helveticus DSM 17754 and recovery by concentration and nanofiltration. The product contains a ferment composition containing the casein-recovered peptides Ile-Pro-Pro and/or Val-Pro-Pro, and living lactic acid bacteria. For the purpose of removing arterial stiffness, the product containing Ile-Pro-Pro and/or Val-Pro-Pro is introduced into an individual. For the purpose of improving vascular elasticity, the product containing Ile-Pro-Pro and/or Val-Pro-Pro is introduced into an individual with using said biologically active peptides.

EFFECT: invention promotes prevention and normalisation of endothelial dysfunction, improves vascular elasticity and removes arterial stiffness.

12 cl, 3 tbl, 5 ex

Description

FIELD OF THE INVENTION

The invention relates to the prevention and treatment of endothelial dysfunction using biologically active peptides and their products. It was found that a product having a high content of short chain peptides is particularly effective for use in accordance with the present invention.

The product used in accordance with the present invention can be manufactured, for example, as a healthy lifestyle food product or a pharmaceutical product. It contains low molecular weight peptides such as Ile-Pro-Pro (IPP), Val-Pro-Pro (VPP) tripeptides or mixtures or concentrates containing them, and acts by improving epithelial function and curing diseases associated with it. A specific objective of the present invention is to reduce stiffness and thus increase blood vessel elasticity using low molecular weight peptides such as Ile-Pro-Pro tripeptides (IPP), Val-Pro-Pro (VPP) or mixtures, concentrates and other products thereof containing.

State of the art

The vascular endothelium locally regulates vascular tone through the release of vasodilating agents such as endothelial relaxing factor (EDRF), nitric oxide, prostacyclin and endothelial hyperpolarizing factor (EDHF), and vasoconstrictors such as thromboxane A ₂ , free radicals and endothelin. Several clinical studies have shown the importance of nitric oxide in both basal and induced vascular tone control in large epicardial coronary arteries and in coronary microcirculation (see review papers by Vapaatalo H, Mervaala E. Clinically important factors influencing endothelial function. Med Sci Monit 2001. ; 7 (5): 1075-1085. Drexler H, Hornig B. Endothelial dysfunction: a novel therapeutic target. J Mol Cell Cardiol 1999; 31: 51-60). The aforementioned relaxing factors of endothelial origin correct endothelial dysfunction, for example, with atherosclerosis. The endothelium is associated with the regulation of vascular tone, modulation of blood coagulation, stimulation and prevention of vascular growth, modulation of inflammation and action as an object of exposure and undesirable effects of drugs. The release of EDRF is stimulated by an increase in the vessels of the inflow, for example, bradykinin, thrombin, acetylcholine and serotonin.

The endothelium controls the tone of the underlying smooth muscles in response to certain pharmacological and physiological stimuli. Functionally, the endothelium is involved in vascular growth, lymphocyte adhesion and immunological regulation, the metabolism of circulating amines, the metabolism of lipoproteins and the integration and transmission of hematopoietic signals.

Endothelial dysfunction is characterized by a decreased secretion of vasodilating mediators, increased production of vasoconstrictor substances, increased sensitivity to vasoconstrictor substances and / or vascular smooth muscle resistance to endothelial vasodilator substances.

Dysfunction is the result of an imbalance between relaxing and contracting factors, or agents that stimulate and inhibit growth. Inflammation, lipoprotein oxidation, or other oxidative stress reactions are factors that influence the development and maintenance of endothelial dysfunction. Theoretically, nitric oxide and its metabolites, as well as cyclic GMF, are the most distinct and direct indicators of endothelial dysfunction.

Endothelial dysfunction can be both a cause and a consequence of several clinical conditions, such as hypertension, atherosclerosis, coronary heart disease, heart failure, diabetes, and high cholesterol in the blood. The most direct evidence of endothelial dysfunction is an imbalance between reduced production or functioning of receptors for vasodilating factors such as nitric oxide, prostacyclin, endothelial hyperpolarizing factor (EDHF), and natriuretic peptides; or increased education or sensitivity to vasoconstrictors, such as endothelin-1, angiotensin II, endoperoxides and thromboxane A ₂ .

Endothelial dysfunction can be treated with several known drugs, the most important of which are angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, nitrate drugs and cholesterol lowering drugs. The effect of ACE inhibitors is mainly based on their ability to improve the action of bradykinin, which enhances the synthesis of nitric oxide in the endothelium. Induced and / or enhanced production of nitric oxide or the addition of nitrates can balance inadequate internal production of nitric oxide. Medications and nitrates act as exogenous EDRF and dilate blood vessels, in addition, they are active as an antithrombotic compound in damaged veins.

In WO 02/34767 A1, Selwood et al., Peptides are described which are fragments of vascular endothelial growth factor (VEGF) and are useful for suppressing angiogenesis. In accordance with the invention, the peptides contain from three to eight amino acids, and a key feature of the amino acid sequence is the presence and location of the main residues, in particular residues of arginine (Arg) and / or lysine (Lys). The document includes a description of several peptides containing from six to sixteen amino acids. None of them contain the sequence Isoleucine-Proline-Proline (IPP) or Valine-Proline-Proline (VPP). According to the publication, peptides may be useful in diseases where angiogenesis plays a significant role in pathology. Such diseases may include diabetic retinopathy, age-related macular degeneration (ARDS), cancer, endometriosis, psoriasis, and rheumatoid arthritis.

WO 99/45941, Sandberg et al., Describe a composition used to improve the ductility, elasticity or appearance of a fabric. The composition is composed of peptides that belong to any of the 41 petid fragments obtained by thermolytic cleavage of elastin. Preferably, the composition comprises a polypeptide having the formula R1-valyl-valyl-prolyl-glutamine-R2, where R1 is the amino terminal portion of the peptide and R2 is the carboxy terminal portion of the peptide. The composition is preferably applied to human skin as part of a cosmetic composition. According to the document, the composition may also be useful in the treatment of hypertension, coronary heart disease, arteriosclerosis, angina pectoris, coronary artery thrombosis, chronic obstructive pulmonary disease and restenosis after angioplastic surgery.

WO 01/91700 A2 and US Pat. No. 6,962,904 B1, Mitts et al., Are based in part on the same studies as WO 99/45941, Sandberg et al. Documents describe compositions for increasing tissue elasticity, and these compositions are composed of peptides corresponding to the sequences found in elastin, in particular, the sequences-Valine-Valine-Proline- and-Valine-Valine-Proline-Asparagin. These compositions are useful for improving the production of elastin in tissues. According to publications, the main practical use is again the use in cosmetics, but it is also indicated that the compositions can be useful in the treatment of, for example, hypertension, coronary heart disease, arteriosclerosis, angina pectoris, coronary artery thrombosis, chronic obstructive pulmonary disease and restenosis after angioplastic operations.

The described peptide fragments of elastin contain a large amount of glycine and / or proline residues compared to other amino acid sequences. Fragments do not include the sequences of Isoleucine-Proline-Proline (IPP) and / or Valine-Proline-Proline (VPP).

The manufacture of biologically active peptides by fermentation has been well studied and described in the prior art, with peptides isolated from milk being of particular interest. It has been shown that they possess, for example, opioid receptor binding ability, ACE inhibitor activity and antimicrobial properties. Some studies have been carried out in connection with, for example, hypertension, but the effects of these peptides on endothelial function have not been described. In particular, the effects of Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP) tripeptides and products containing them on endothelial function have not been described in the prior art.

Tripeptides VPP and IPP are known compounds that have been described as ACE inhibitors having antihypertensive effect (lowering blood pressure). For example, in J Dairy Sci 78 (1995) 777-783, Nakamura et al. Describe the use of a starter culture containing Lactobacillus helveticus and Saccharomyces cerevisiae to produce two ACE inhibitors. Both compounds have been identified as tripeptides, Val-Pro-Pro and Ile-Pro-Pro. Although this publication does not describe the antihypertensive effect of tripeptides in vivo , it is envisaged that this is the next goal of research.

US Pat. No. 5,449,661 to Nakamura et al. Discloses the preparation of a peptide comprising the Val-Pro-Pro tripeptide sequence and its use for lowering high blood pressure. The peptide was obtained by fermentation of skim milk powder using strain JCM-1004 Lactobacillus helveticus , after which the peptide is purified chromatographically and lyophilized.

In WO 99/16862, Yamamoto et al. Describe a strain CM4, FERM BP-6060 of Lactobacillus helveticus , which is capable of producing large quantities of Val-Pro-Pro and / or Ile-Pro-Pro tripeptides.

WO 01/32905, Valio, describes a product having antihypertensive properties and its preparation. The product is produced by fermentation of a casein-containing feed using lactic acid bacteria, and nanofiltration of the resulting fermentation product containing the peptide. The antihypertensive properties of the product are manifested, in particular, due to the content of IPP and VPP tripeptides in it. WO 03/070267, Valio, describes the use of IPP and VPP, as well as the product described in WO 01/32905, in the preparation of a product that increases the availability of minerals. This product can be used, for example, to enhance osteogenesis (bone formation), strengthen the skeletal system and treat or prevent osteoporosis.

Blood vessels are involved in diseases associated with elastic properties, including hypertension, arteriosclerosis, angina, angiogenesis, myocardial infarction, coronary artery thrombosis, restenosis after angioplastic surgery, and chronic obstructive pulmonary disease. Cardiovascular diseases are one of the most common diseases in the world, and they are on the list of the five most life-threatening diseases in many countries. Unfortunately, with an increase in living standards, the risk of these diseases also increases, and, therefore, they will be even more important in the future. Currently, in addition to conventional medicines, the consumer is offered an attractive alternative in the form of functional products.

Functional products that improve the elasticity of blood vessels and improve and normalize the function of the endothelium will thus be welcomed as part of a regular diet. In addition, the introduction of peptides with useful properties in relation to arterial stiffness is worthy of consideration as a medical or pharmaceutical product.

SUMMARY OF THE INVENTION

Thus, it is an object of the present invention to provide a product that, as part of a regular diet, or drug and pharmaceutical product, improves or normalizes endothelial function and, therefore, is able to prevent, alleviate or cure disorders and diseases associated with endothelial dysfunction. Endothelial dysfunction plays a significant role in determining the stiffness or elasticity of blood vessels, which, in turn, is very important for many serious diseases, for example, coronary heart disease, arteriosclerosis, angina pectoris, coronary artery thrombosis, chronic obstructive pulmonary disease and restenosis after angioplastic operations. Therefore, the ability to increase the elasticity of blood vessels is a particularly important property of the present invention.

The product to be used according to the invention consists of or contains peptides that improve endothelial function.

The next objective of the present invention is the provision of the product, as such, as an agent that improves endothelial function, or for use in the manufacture of functional foods or drugs intended for consumption.

Another objective of the present invention is the provision of a method of preventing, alleviating or treating endothelial dysfunction, as well as disorders and diseases associated with it, by administering to an individual in need of such treatment, peptides that improve endothelial function, or a product containing them sufficient to achieve the desired effect of quantity.

Another objective of the present invention is the provision of a method of preventing, alleviating or treating endothelial dysfunction, as well as disorders and diseases associated with it, by administering to an individual a product with a high content of low molecular weight peptides derived from casein and obtained by fermentation of casein-containing feedstock using strain LBK-16H, DSM 13137 Lactobacillus helveticus, or strain LB 1936, DSM 17754 Lactobacillus helveticus, and optionally a partial or complete removal of casein and / or other proteins Molo a and / or lactose, in a sufficient to achieve the desired effect amount. In a preferred embodiment, the fermentation product also undergoes nanofiltration.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the present invention, it was surprisingly found that the above objectives are achieved using low molecular weight peptides, in particular, Ile-Pro-Pro tripeptides (IPP), Val-Pro-Pro (VPP). It has now been proven that these peptides are able to normalize endothelial functions, improving the elasticity of blood vessels and counteract arterial stiffness.

The peptides used in accordance with the present invention are biologically active peptides corresponding to those obtained by hydrolysis of casein or casein-containing raw materials such as milk. Short chain peptides di-, tri- and tetrapeptides and mixtures thereof are considered suitable. In particular, the peptides are IPP and VPP tripeptides, mixtures of peptides including IPP and VPP, and products and compositions containing them.

The preferred embodiment provides excellent opportunities for normalizing endothelial dysfunction and endothelial-related disorders.

Thus, the invention relates to the use of low molecular weight peptides isolated from casein for the manufacture of a product that improves endothelial function.

The invention further relates to the use of a product containing low molecular weight peptides isolated from casein for the manufacture of a product that improves endothelial function.

Preferably, the low molecular weight casein-derived peptides include di-, tri- and tetrapeptides and mixtures thereof. Most preferably, a high content of IPP and VPP.

Further, the invention relates to the use of a product with a high content of low molecular weight casein-derived peptides that were obtained by fermentation of a casein-containing feed using lactic acid bacteria, and possibly nanofiltration of the resulting peptide-containing fermentation product, in the manufacture of a final product that improves endothelial function.

As a preferred embodiment, the invention relates to the use of a starter composition which contains peptides isolated from casein, minerals and live lactic acid bacteria for the manufacture of an endothelial function improving product.

The invention also relates to a method for normalizing endothelial function and endothelial-dependent disorders by administering to an individual peptides that normalize endothelial function and endothelial-dependent disorders, or a product containing them in an amount sufficient to achieve the desired effect.

First of all, the individual is a person. The beneficial effects of using the products according to the invention are naturally also beneficial for animals, especially domestic animals and farm animals. Examples thereof include dogs, cats, rabbits, horses, cows, pigs, goats, sheep and birds.

Tripeptides VPP and IPP can be obtained synthetically or by hydrolysis of a feed containing these sequences. In a preferred embodiment, the peptides are derived from casein or casein-containing feedstock by fermentation. Particularly preferred is the use of the method described in WO 01/32905, where a product containing biologically active peptides is obtained by fermentation followed by concentration and nanofiltration. This preferred embodiment provides excellent opportunities for the use of feedstocks of various types and modifications of the composition of the final product according to the requirements, as described in detail in WO 01/32905, hereby incorporated by reference.

In the case where the biologically active peptides for use in accordance with the present invention are obtained by fermentation, the feedstock may be any product containing sequences of the desired biologically active peptides as part of its own peptide or protein sequence. Milk protein, especially casein, is preferred for use as such or in the form of various preparations. Suitable types of raw materials also include various dairy products containing casein, such as skim milk or milk with different fat content as such, or in the form of appropriate milk powder and lactic acid products such as sour milk, buttermilk, yogurt, yogurt, unripened cheese and etc.

Fermentation can be carried out using any lactic acid bacteria that are capable of producing the desired peptides from the feedstock. Suitable lactic acid bacteria can be found among species belonging to the genus, for example, Lactobacillus, Lactococcus, Leuconostoc, Streptococcus and Bifidobacterium . The most pronounced among lactic acid bacteria proteolytic properties possesses Lactobacillus helveticus , and, therefore, is considered as the most suitable for this purpose. A preferred strain of Lactobacillus helveticus is L.helveticus LBK-16H, DSM 13137, which is described in detail in patent publication WO 01/32836. Particularly preferred is Lactobacillus helveticus LB 1936, DSM 17754, which is described in detail in patent publication FI20065054.

Lactic acid bacteria can be used as pure cultures or mixed cultures, separately or in conjunction with commonly used and commercially available starter cultures. Lactic acid bacteria can also be used in conjunction with other microorganisms.

Fermentation conditions are selected in such a way as to satisfy the requirements of the strain used to form a sufficient number of biologically active peptides to obtain the desired effect. The selection of suitable conditions, such as temperature, pH and aeration, is part of the practical knowledge of a person skilled in the art. Typically, fermentation is carried out at a temperature of about 30 to 45 ° C. Fermentation is carried out until the required amount of biologically active peptides is formed. Typically, the process takes about 20 to 30 hours. During fermentation, a mixture of various peptides is formed. In the case of sufficiently long fermentation, relatively short di- and tripeptides, such as Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP), are generally obtained. Preferably, the incubation time is from about 22 to 24 hours.

The resulting cell suspension can be used as such, or the peptides can be separated and purified using standard techniques. A preferred type of treatment is the concentration, for example, by evaporation or partial or complete drying of the cell suspension by spreading the suspension over the surface of the plate, drying and finally grinding into a well-preserved dry powder.

Additional processing of the cell suspension by nanofiltration is an excellent way to facilitate the production of peptides (active against endothelial functions, such as arterial stiffness) together with the necessary composition of minerals in the same product. By choosing the type of membrane for nanofiltration and process conditions, it is possible to influence the composition of the resulting product.

Nanofiltration is carried out until the desired dry matter content is reached, which can be from about 20 to 40%, or to an approximate ratio of volume concentrations of 5 to 20. By nanofiltration, the peptide content in the product increases. The composition of the final products naturally depends on the fermentation conditions used, optional processing by nanofiltration and other possible preliminary and additional treatments.

Surprisingly, in accordance with the present invention, IPP and VPP tripeptides and products containing them reduce arterial stiffness and are thus useful in arterial dysfunctions. During an exposure period of ten weeks, the ambulatory arterial stiffness index (AASI), measuring arterial stiffness, significantly improved in the group receiving the product containing IPP and VPP tripeptides compared to the control group; even in the first non-optimized test, the index decreased by about 9% compared with the control group. The average AASI at the end of the treatment period was 0.31 ± 0.15 in the Lactobacillus helveticus group and 0.34 ± 0.17 in the control group.

The products described above can be used as such to achieve the desired effect. The products can also be dried and used in the form of a powder or lyophilized preparation. The products are preferably also used in the preparation of functional foods or other products. Application as medicinal or pharmaceutical products is also possible.

The term “food” is broadly defined in the present invention, encompassing all consumable products that may be in solid, jelly, or liquid form, and encompassing both finished products and products to which are added as an additive or part of the product during consumption biologically active peptides or products containing them. Food products may, for example, be products of the dairy industry or the beverage industry. Typical products include dairy products such as yogurt, yogurt, cottage cheese, sour milk, sour whole milk, buttermilk, other dairy products, unripened cheeses and mature cheeses, snack fillings, etc. Another important group includes drinks, such as whey, fruit drinks, and carbonated drinks.

In accordance with the invention, biologically active peptides or products containing them are used in sufficient amounts to achieve the desired effect. In the case of using the product obtained by the two-stage method described above, the amount for use mainly depends on the degree of concentration of the serum and is, for example, from 0.1 to 30%, preferably from about 5 to 15%, when calculated relative to the weight of the final product .

Biologically active peptides or products containing them can be added to the food product during its manufacture or to the finished food product. The food products contemplated thus include the desired peptides or a product containing them, in addition to the other components contained in the respective food products, and are fully consistent in taste and characteristics with these conventional products.

The invention will be described in detail through the following examples. These examples are provided only to illustrate the invention and should not be construed as limiting in any way the scope of protection. The examples set forth demonstrate the beneficial, beneficial and therapeutic effects of IPP and VPP, either alone or in combination, on arterial stiffness by enhancing endothelial function, as well as preferred end products for receiving biologically active peptides.

EXAMPLE 1

The effect of biologically active product on arterial

rigidity

Arterial stiffness can be measured using ultrasound equipment or applanation tonometry, or using the ambulatory arterial stiffness index (AASI). AASI is defined as 1 minus the tangent of the angle of inclination to the x-axis of diastolic or systolic pressure during 24-hour outpatient monitoring, and is a reliable indicator of arterial stiffness (Li Y, Wang JG, Dolan E, Gao PJ, Guo HF, Nawrot T. Ambulatory arterial stiffness index derived from 24-hour ambulatory blood pressure monitoring. Hypertension 2006; 47 (3): 359-64).

In a randomized trial where the control group took a placebo at the same time, 94 patients with high blood pressure who did not receive any drugs took either milk fermented with the Lactobacillus helveticus LBK-16H strain in accordance with Example 2 or the control product within ten weeks after four-week preparatory period. A twenty-four hour outpatient blood pressure measurement was performed at the beginning and at the end of the experimental period. Mean baseline values of systolic and diastolic blood pressure were 132.6 ± 9.9 / 83.0 ± 8.0 mm Hg. in the group taking Lactobacillus helveticus , and 130.3 ± 9.6 / 80.2 ± 7.0 mm Hg. in the control group.

At the end of the ten-week experimental period, systolic and diastolic blood pressure decreased to a greater extent in the group receiving the product Lactobacillus helveticus , compared with the control group. The treatment effect on SBP (systolic blood pressure) was -4.1 (95% Cl: -6.6 to -1.8, p <0.001). The treatment effect on DBP (diastolic blood pressure) was -1.8 (95% Cl: -3.7 to -0.0, p = 0.048).

The AASI index was determined using the values of ambulatory blood pressure and their analysis by the method proposed by Li et al., Ibid. The AASI index increases linearly with age in both men and women. Low values of the AASI index are more favorable than high values, and therefore, if the value decreases, the arterial stiffness index improves. The average AASI baseline values were 0.36 ± 0.15 in the Lactobacillus helveticus group and 0.36 ± 0.17 in the control group. At the end of the experimental period, the average AASI values were 0.31 ± 0.14 in the Lactobacillus helveticus group and 0.34 ± 0.15 in the control group.

During the ten weeks of the experimental period, the AASI index improved statistically significantly in the tested groups (p = 0.043), but not statistically significant in the control group (p = 0.47) (Table 1).

Table 1 AASI Index Changes After a 10-Week Pilot Period Relevance
(2 sided) Average difference Test group AASI Index Changes 0,043 -0.0427 Control group AASI Index Changes 0.474 -0.0195

EXAMPLE 2

Obtaining products suitable in the present invention

The strain Lactobacillus helveticus LBK 16-H, DSM 13137, was grown in MRS at 37 ° C for 24 hours and plated in reconstituted milk (10%) to form a starter culture. After two growth cycles, the leaven (15%) was added to the fermenter medium prepared from 9-10% skimmed milk powder and sterilized at 110 ° C for 10 minutes. Fermentation was carried out at 37 ° C for 22 to 24 hours with continuous strong stirring. The product (a) can be used as such, in dry and / or ground form, or the desired peptides can be separated from the product using known methods.

The procedure was repeated using Lactobacillus helveticus LB 1936, DSM 17754, and whole milk or buttermilk, respectively, instead of skimmed milk powder, in order to produce the corresponding products b and c.

EXAMPLE 3

Obtaining the final product suitable in the present invention

Peptide Concentrate

Sour milk containing peptides active in correcting endothelial dysfunction was obtained by adding about 1% of the peptide concentrate to commercially available sour milk. The composition of the product is shown in Table 2, which also shows, for comparison, the composition of a commercially available fermented milk product, sour milk AB, manufactured by Valio Ltd.

To obtain the Evolus® peptide concentrate, fermentation was carried out as described in Example 2. The resulting cell suspension was separated using a clarification centrifuge. Thus, the pre-treated serum was nanofiltered through a Nanomax-50 membrane at 40 ° C and a pressure of 30 bar. Serum was filtered until the volume concentration ratio reached 9.

table 2 The composition of the product Evolus and commercial lactic acid
product (sour milk AB, Valio Ltd) Nutritional Information
product / 100 g (portion 200 g) Evolus Sour milk AB Calorie content 260 kj /
61 kcal 220 kJ /
53 kcal Squirrels 3.0 g 3.2 g Carbohydrates 12 g 4.4 g Of which lactose is not more 1.0 g * Cellulose 0 g 0 g Fats 0.1 g 2.5 g Calcium 150 mg ** 120 mg Potassium 233 mg 170 mg Magnesium 13 mg 11 mg Sodium 24 mg 39 mg Tripeptides Ile-pro-pro 1.1 mg 0 Val-pro-pro 1.3 mg 0 * Over 80% of lactose is enzymatically destroyed
** 25% of recommended daily intake

EXAMPLE 4

Obtaining the final product suitable in the present invention

Fermented milk drink Evolus Tehojuoma

To obtain a daily dose of Evolus® Tehojuoma fermented milk drink, fermentation was performed as described in Example 2. Next, 1% Evolus® peptide concentrate prepared as described in Example 3 was added to the fermented milk product. The product containing the required daily dose of the active ingredient as such is intended for consumption.

The nutritional information of the product is shown in Table 3. Other ingredients that contain a fermented milk drink are pasteurized skim milk, milk protein concentrate, thickeners, seaweed calcium, acidity regulator, yeast, flavoring, vitamins (folic acid, B6, B12) and coloring .

Table 3 Evolus® Tehojuoma Lactic Drink Composition Nutritional Information / 100 g (100 ml serving) Evolus Calorie content 260 kj /
61 kcal Squirrels 3.5 g Carbohydrates 11 g Of which lactose is not more 1.0 g * Cellulose 0 g Fats 0.1 g Calcium 200 mg ** Potassium 410 mg Magnesium 15.5 mg Sodium 38 mg Folic acid 66 mcg *** Vitamin B6 0.66 mcg *** Vitamin B12 0.33 mcg *** * Over 80% of lactose is enzymatically destroyed
** 25% of recommended daily intake
*** 33% of recommended daily intake

EXAMPLE 5

Obtaining the final product suitable in the present invention

Soft cheese

Soft cheese, such as fresh cheese, cream cheese and homemade cheese (pressed cottage cheese) was obtained according to generally accepted production technologies. For example, Evolus® processed cheese was prepared by mixing cottage cheese, cream or butter and other ingredients, cooked and packaged. The Evolus® peptide concentrate prepared as described in Example 3 was added to the curd-rich mixture. A product containing the required daily intake of the active ingredient, as such, is intended for consumption.

Claims (12)

1. The use of Ile-Pro-Pro and / or Val-Pro-Pro to obtain a product that reduces arterial stiffness. 2. The use of Ile-Pro-Pro and / or Val-Pro-Pro to obtain a product that improves the elasticity of blood vessels. 3. The use according to claim 1 or 2, characterized in that Ile-Pro-Pro and / or Val-Pro-Pro are in the form of a food product. 4. The use according to claim 3, characterized in that the food product has a high content of low molecular weight peptides isolated from casein, and which is obtained by fermentation of casein-containing feedstock with lactic acid bacteria. 5. The use according to claim 4, characterized in that the food product has a high content of low molecular weight peptides isolated from casein, and which is obtained by fermentation of casein-containing feedstock with a strain of Lactobacillus helveticus LBK-16H, DSM 13137, or with a strain of Lactobacillus helveticus LB 1936, DSM 17754, optionally with partial or complete removal of casein and / or other milk and / or lactose proteins and nanofiltration of the resulting protein-containing fermented product. 6. The use according to claim 4, characterized in that the product comprises a yeast composition containing low molecular weight Ile-Pro-Pro and / or Val-Pro-Pro peptides and live lactic acid bacteria, isolated from casein. 7. The use according to claim 3, characterized in that the food product is a product of the dairy industry, the beverage industry, the processed food industry, the finished meat dishes industry, the baking industry or the confectionery industry. 8. The use according to claim 7, characterized in that the product is a fermented dairy product. 9. The use according to claim 7, characterized in that the product is a drink, preferably a whey drink, fruit drink or beer. 10. The use according to claim 1 or 2, characterized in that the product is a product of the pharmaceutical industry. 11. A method of reducing arterial stiffness, comprising administering to an individual a product comprising Ile-Pro-Pro and / or Val-Pro-Pro as defined in claim 1. 12. A method for improving the elasticity of arterial vessels, comprising administering to an individual a product comprising Ile-Pro-Pro and / or Val-Pro-Pro, as defined in claim 2.