Method for teacheability and memory improvement in mammals

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely for teacheability and memory improvement in mammals. An infant takes a milk formula containing casein glycomacropeptide in amount sufficient to ensure introducing at least 100 mg/kg of sialic acid a day.

EFFECT: method extends the range of products for teacheability and memory improvement in an infant.

11 cl, 14 tbl, 13 dwg, 7 ex

 

The technical field

The present invention relates to a method for improving learning and/or memory in mammals using diet and, in more detail, to a method for improving learning and/or memory in mammals by introduction of a food source of sialic acid.

The level of technology

It was long ago noticed that breastfeeding is the preferred method of feeding for a person. Among the most significant advantages, it should be noted optimal mental development. There is therefore a need to create a composition formula, which will provide optimal mental development of the child on artificial feeding, comparable in pace with the development of children breastfed.

Compared to the brain of other primates, the brain of an adult four times as much relative to their body mass, however, the man-cub at birth is relatively immature. Parker. S.T. et al., Origin of Intelligence, pp.313-345, The Johns Hopkins Univ. Press, Baltimore, MD (1999). The brain is actively developing both before and after birth, for its proper development requires the timely supply of nutrients. Optimal nutritional support during critical phases of early brain development has a lasting effect on cognitive function is - the object of attention of doctors and clinicians. Lucas, A. et al., BMJ, 217:1481-1487 (1998), and Lucas. A. et al., Lancet, 339:261-264 (1992). In a randomized double-blind controlled studies in premature infants has been shown that IQ at 8 years (IQ) in children treated with standard infant formula, was lower than in children who received fortified infant formula. Cm. Lucas, Id. (1998). Of concern is the fact that almost half of the male children treated with standard formulas, noted reduced IQ, constituting only 13% of the IQ of children, fed enriched formula milk. Such studies emphasize the special susceptibility of premature infants and children with low body weight at birth to optimal breastfeeding.

Specific unique components of human milk have the ability to support the rapid development of the brain. In particular, sialic acid (the family of N - and O-substituted derivatives of neuraminic acid; and, in particular, N-acetylneuraminic acid, or NANA), present in large quantities in human milk oligosaccharides (up to 1 g/l) form of the limit functional residues of gangliosides and glycoproteins brain. Wang, W. et al., Am. J. Clin. Nutr., 74:510-515 (2001) and Carlson, S. E, Am. J. Clin. Nutr., 41:720-726 (1985). Components of sialic acids in high to the concentrations present in the breast milk of the mother immediately after birth. Idota et al., in J. of Japanese Soc. Of Nutr. and Food Sci. (Nihon Eiyo Shokuryo Gakkai-shi), 47(5):363-367(1994), demonstrated a significant decrease in the content of the 6'-sialyllactose in human breast milk from 3 to 482 day after birth.

In nature the highest concentration of sialic acid are found in the cerebral cortex of man. Schauer, R., Sialic acids, Chemistry, Metabolism and Function, Springer-Verlag, Wien, New York (1982) and Svennerholm, L. et al., Biochem Biophys Acta, 1005:109-117 (1989). In particular, neural cell adhesion molecule (NCAM) is validierung protein, which apparently plays a very important role in the processes of learning and memory. He participates in many morphogenetic processes, including cell migration, neurite outgrowth, pathfinding sprouting of nerve fibers, the sprouting of nerve fibers, regeneration and synaptic plasticity. Nakayama, J. et al., Virchows Archiv, 433:419-426 (1998), Mahal, L, K. et al., J.Biol. Chem., 277:9255-9261 (2002), and Ong, E. et al., Glycobiology, 8:415-424 (1998). In rodents level of polysilicate NCAM directly associated with an enhanced ability to learn and remember. Cremer, H. et al., Nature, 367:455-459 (1994).

Researchers have shown that a large part of the N-acetylneuraminic acid (NANA) in the brain newborn of rats treated with NANA by intraperitoneal introduction, included in intrasynaptic faction. Morgan, B.L. G. et al., Br. J.Nutr., 46:231-238 (1981). Later, Carlson, S.E. et al., in J.Nutr., 116:881-886 (1986) showed that the oral, and intraperitoneal N-acetylneuraminic acids leads to the formation of many more cerebral and cerebellar of gangliosides and glycoproteins N-acetylneuraminic acids than glucose.

Morgan, C. L. G. et al., in J. Nutr., 110:416-424 (1980) also showed that rat pups fed a poor diet, which was administered intraperitoneally injected N-acetylneuraminic acid, quickly learned and memorized the maze than calves treated with glucose, in addition, they recorded fewer behavioural problems, in spite of poor nutrition.

Composition based on cow's milk usually contain little of sialic acids. In one study it was shown that the concentration of sialic acids in several compositions containing a combination of casein/whey) was less than 200 mg of sialic acid per liter. In addition, the composition of soy proteins also contain a very low amount of sialic acids, even in comparison with compositions based on cow's milk. Thus, the composition of soy protein that does not contain lactose, have a very low content of sialic acids. Some studies have shown that dietary supplements of sialic acids or containing sialic acid, have a significant advantage.

There are several sources with elovich acids in their various conjugated forms. These include, without limitation specified free N-acetylneuraminic acid (or sialic acid), oligosaccharide of sialyllactose, gangliosides containing sialic acid, and macropeptide casein (CMP), also known as glycomacropeptide (GMP) and, if it is isolated from cow's milk, glycomacropeptide casein (CGMP), etc.

The method of obtaining CGMP described in application for U.S. patent 20040022918, which is to create cheese from milk by coagulation of cow's milk, with chymosin causes stvorazhivanii of coagulate, as he squeezes the serum. Macropeptide casein (CMP) is released from the protein casein in the feedback himopsina on Kappa-casein, the whey is removed about 90% of the SMR. IDA is a heterogeneous group of proteins that includes all genetic variation and posttranslational modifications of Kappa-casein (Yvon et al., Reprod. Nutr. Dev., 34:527-537 (1994)). The predominant hydrocarbon is sialic acid. Glycomacropeptide or GMP is the main (50-75%) component of the CMP. The hydrocarbon component of the GMP makes it soluble in 12% solution of trichloroacetic acid. A number of analytical measurements includes preliminary treatment, which includes the TCA solution that allows you to allocate at least part replicationmanager IDA. For example, the method, published in The Offiial Journal of the European Communities (L228/10 Annex IV), includes the use of HPLC for measurement of the GMP diet products and level of GMP to calculate the amount of cheese whey present in the sample. Other methods of obtaining CGMP of milk described Brody, E.P., in., Br. J. of Nutr 84 (Suppl.1):S39-S46 (2000).

How to add sialic acid or sources of sialic acid in certain nutrient mixture described in U.S. patent No. 6,506,422, in which are disclosed certain nutrient mixture containing glycomacropeptide casein and complementary essential amino acids except phenylalanine, designed for patients with phenylketonuria. The levels of sialic acid in milk formulas are not mentioned.

In U.S. patent No. 6,270,827 describes compositions containing human milk proteins or recombinant factors of sustainability of the owner, one of which is a recombinant human Kappa-casein, for the enrichment of artificial milk formulas.

In U.S. patent No. 4,762,822 discloses the use of N-acetylneuraminic acid or gangliosides containing sialic acid in milk mixtures to protect the newborn from microorganisms that cause infections of the gastrointestinal tract.

International patent application WO 01/60346 A2 for the nutrient mixture containing oligosaccharides of oligofructose and sialyllactose as prebiotic additives to improve the Oia growth of bifidobacteria in the intestine, which can be used in combination with milk mixture.

The application WO-A-00 describes the use of a hydrolysate of milk proteins for the treatment of diseases of bones and teeth. Glycomacropeptide casein (CGMP) extracted from fresh whey by a combination of electrodialysis, cation exchange resin, anion exchange resin, evaporation, spray-drying, ultrafiltration and lyophilization, and used for food fortification and liquid enteral compositions.

Although it was shown that the use of sialic acid for various purposes, there is no evidence that dietary supplements sialic acid affect learning and memory in mammals. Since the use of various dietary supplements is a simple and common way of introducing agents into the organism, especially a newborn mammal, it can be useful to improve learning and memory in mammals. This method is particularly useful for newborn mammals in the neonatal period, when there is a need to improve learning and memory.

BRIEF description of the INVENTION

Briefly, the present invention relates to a new method of improving learning and/or memory in mammals, which consists in the introduction to the mammal glycomic is peptide casein in amount, sufficient to improve memory and/or learning in mammals.

The present invention also relates to a new method of increasing the number of peptide linkages of sialic acid in the brain, which consists in the introduction to the mammal the number of glycomacropeptide casein, which is sufficient to increase the number of peptide linkages of sialic acid in the mammalian brain.

The present invention relates to a new method of improving learning and/or memory in mammals, which comprises the following steps: determining whether a mammal in need of improved memory and/or learning; and, if needed, the introduction of glycomacropeptide casein in an amount sufficient to improve the processes of memory and/or learning in mammals.

This invention has many advantages, among which may be noted that this invention provides a method of improving learning and/or memory in mammals due to dietary supplements, as well as the method that is particularly useful for mammalian subjects in the neonatal period, which need improvement of learning and memory.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 presents a schematic illustration of a maze with 8 compartments, which is useful for testing abusement and memory in piglets.

Figure 2 presents a schematic illustration of one of the compartments of the maze presented in figure 1

Figure 3 presents a visual signals used in the job training 1 and (C) job training 2, when testing learning in pigs.

Figure 4 presents a graph showing the number of piglets in each of the four groups learned the correct response to a visual cue in the task 1 (the number of animals represented as a function of the number of experiments), while pigs from Group 4 (2nd best) received a diet containing sialic acid in the amount of 842 mg/l, enriched glycomacropeptides casein (240 mg/kg / day power acid), pigs from Group 3 (3rd best result) received a diet containing sialic acid in the amount of 600 mg/l, enriched with glycomacropeptide casein (171 mg/kg / day sialic acid), piglets from Group 2 (the best result) received a diet containing sialic acid in the amount of 250 mg/l, enriched glycomacropeptides casein (71 mg/kg / day sialic acid), and pigs from Group 1 (the worst result) received a diet enriched in sialic acid in the amount of 77 mg/l (25 mg/kg / day sialic acid) without glycomacropeptide casein.

Figure 5 presents a graph showing the number of piglets in each of the four groups easy is or the correct response to a visual cue in the task 2 (the number of animals represented as a function of the number of experiments), while pigs from Group 4 (2nd best) received a diet containing sialic acid in the amount of 842 mg/l, enriched glycomacropeptides casein (240 mg/kg / day power acid), pigs from Group 3 (3rd best result) received a diet containing sialic acid in the amount of 600 mg/l, enriched glycomacropeptides casein (171 mg/kg / day sialic acid), piglets from Group 2 (the best result) received a diet containing sialic acid in the amount of 250 mg/l, enriched with glycomacropeptide casein (71 mg/kg / day sialic acid), and pigs from Group 1 (the worst result) received a diet enriched in sialic acid in the amount of 77 mg/l (25 mg/kg / day sialic acid) without glycomacropeptide casein.

Figure 6 presents a bar chart showing the total number of errors 1 and 2 tasks in each group of piglets, while the chart shows that the greatest number of errors were made by the piglets from the group who did not receive enrichment glycomacropeptides casein and diet support in any quantity leads to improved learning animals.

Figure 7 shows two bar charts, this chart (A) shows the average number of errors committed by animals from each of the four groups in the job memory 1, and diagram (C) shows the em average number of errors, perfect animals from each of the four groups in the job memory 2.

On FIG presents two bar chart, with the chart (A) represents the comparison of the levels of cortisol in the plasma in each of the four groups of animals for each week of the study; and diagram (C) represents the comparison of the levels of cortisol in different groups receiving combined treatment and control group each week 5-week trial.

Figure 9 presents a bar chart, which compares the concentration of sialic acid in the gray matter of the brain of animals from the experimental and control groups, and the concentration of sialic acid in the gray matter of the brain is higher in animals fed a diet enriched with glycomacropeptide casein.

Figure 10 presents two scatter plot of concentration-related protein sialic acid in the tissue of the frontal cortex depending on the number of experiments needed to piglets learned visual cue in the task 1 and task 2 (B), both charts show that the highest levels associated with protein sialic acid in the tissue of the frontal cortex correlated with improved memory in piglets.

Figure 11 presents two that is Ichnya chart concentration-related glycoside of sialic acid in the tissue of the frontal cortex depending on the number of experiments, it took to the pigs learned visual cue in the task 1 and task 2 (B), both charts show that the highest levels associated with protein sialic acid in the tissue of the frontal cortex correlated with improved memory in piglets.

On FIG presents a graph showing the number of errors in the job memory 1 in Group 4, depending on the concentration associated with protein sialic acid in the tissue of the frontal cortex, from the graph we see that the animals with the highest level associated with protein sialic acid in the tissue of the frontal cortex, make fewer errors in tasks of memory and demonstrate the best ability to memorize than animals in which the level of sialic acid below.

On FIG presents a graph showing the average weight gain of piglets from each of the four groups during the whole study, the chart shows that the diet in each group was comparable to the expected excess nutritional requirements of pigs.

DETAILED DESCRIPTION of PREFERRED embodiments of the INVENTION

In accordance with the present invention it has been shown that learning and memory in mammals can be stimulated by the introduction of mammals of glycomacropeptides in number, sufficient to improve memory and/or learning in mammals. According to a useful variant of implementation of the present invention glycomacropeptide casein may be included in the composition.

It was shown that pigs fed a diet containing 77 mg/l of sialic acid (without enrichment) and 250 mg/l 600 mg/l and 842 mg/l of sialic acid-enriched glycomacropeptides casein, demonstrate different abilities to memorize and learn. In fact, it was shown that as the process memory and the learning process is improved in animals fed a diet enriched with glycomacropeptide casein, compared to animals not receiving enriched with casein, but otherwise, completely balanced nutritional diet.

This improvement was unexpected, since the authors of the invention have not yet seen evidence that it is possible with the help of dietary supplements or diets enriched with glycomacropeptide casein, improve memory and learning ability. Also, previous studies have demonstrated a correlation between reduced food intake and the inclusion of CGMP in the diet. See, for example, U.S. patent No. 20040077530 and 20030059496. Because reduced food intake is often associated with low mental development and does not contribute to the improvement of learning is memory, the opening made by the authors of the present invention, that the inclusion in the diet of CGMP can significantly improve these indicators, was unexpected.

The authors of the present invention found that glycomacropeptide casein is extremely useful dietary Supplement to improve memory and learning. This is surprising, since cow's milk, in General, has a low content of sialic acid, in addition, macromolecule that binds sialic acid differ from macromolecules human milk. However, the authors of the present invention showed that glycomacropeptide casein is surprisingly useful dietary Supplement for improving memory and learning, and, in addition, CGMP is relatively inexpensive, readily available and safe for children Supplement.

The present invention is useful for any mammal. However, it is most useful for a person. The mammal can be any age. However, it was shown that the claimed method is most useful for mammals at the age of about 1 day to about 4 years. This includes age newborns and children of early age, if we are talking about man. Preferably, the mammal is in the neonatal period of development. Used herein, the term "neonatal" IP is alzueta to describe age from birth to 2 years of life.

Method declared in accordance with the present invention is most useful in the case when a mammal in need of improved memory and/or learning. The term "in need of improved memory and/or learning" is used here to describe the entity that will benefit, no matter big or not, from the way used to improve memory and/or learning. This method can include the following steps: to establish whether a mammal in improving learning and/or memory; and, if needed, to enter the mammal a composition comprising an effective amount of glycomacropeptide casein.

The term "improving learning/learning processes" means any improvement, no matter whether significant or not, the ability to learn the subject. The term "improve memory/memory processes" means any improvement, no matter whether significant or not, the ability to memorize the subject. Improving learning and/or memory can be measured using one or more tests used to assess learning and/or memory in children. Examples of such tests include, without limitation specified, the test pagana to define intelligence in children (FTII), target permanent scale of Uzgiris-hunt in the modification of Dunst (PAS)test cross-modal transfer is (CMT), the Bayley scales for assessing development in children, second edition, the mental development index (MDI), questionnaire Bailey to assess the development of the nervous system in children (BINS) and neurological test Amiela-Tisone (AT). For more information on testing processes of learning and memory in children can be obtained from Bayley, N., Manual for the Bayley scales for infant development. Psychological Corporation, New York (1969); Bates, J.E. et al., Child Dev., 50:794-803 (1979); and Black, M.M. et al., Bayley Scales of Infant Development II Assessment. Unlimited Learning Resources, Winston-Salem, NC(2003).

It is believed that the need to improve the processes of learning and/or memory in mammals and in particular humans can be caused by many factors that are well known to specialists in this field. For example, these factors include malnutrition, various environmental factors such as exposure to certain metals, oxygen deficiency, injury, disease and other factors that determine the need for such improvement. As discussed above, the field of technology it is clear that there is a connection between the intellectual abilities of mammals in the neonatal period and the level of certain nutrients in the diet, it was shown that sialic acid is one such nutrient. Thus, in accordance with the present invention is disclosed need the diamonds in improving learning and/or memory or the method of determining the need for improvements in teaching and learning and/or memory in a mammal; in this case, at least part of the nutritional needs of a mammal is satisfied by introducing a composition containing at least 100 mg/l of sialic acid, and, in particular, almost all the nutritional needs of a mammal are satisfied by introducing a composition containing less than 100 mg/l of sialic acid.

Used when the expression is satisfied, at least part of the nutritional needs of a mammal"is meant that at least 25% of the food needs of a mammal is satisfied claimed composition in most part of the period of time from birth until the present age. Preferably, a specified part of the food needs of a mammal, which met over a longer period of time from birth until the present age, is at least 50%, more preferably 75%, and more preferably, is almost all the nutritional needs of a mammal.

Because the proteins in soy and cow's milk proteins the number of sialic acid is small, especially compared with human milk, particularly in colostrum and milk, produced in the early stages of lactation, it is believed that the mammal receiving mol is cnie mixture, in which a large part of the protein composition comprises the protein of soy or cow's milk with an average sialic acid of less than 100 mg/l, requires improvement of the processes of learning and/or memory. In particular, it is considered that such a case is when at least 75% of the total weight of protein in the composition presents soy protein or proteins in cow's milk, and this is even more true, than when the whole protein composition formula represented soy protein or proteins in cow's milk.

In fact, it is considered that the need to improve learning and memory in mammals occurs when the nutritional needs of a mammal are satisfied by the diet, to ensure the delivery of sialic acid in a quantity that is lower than the number, which normally enters the body of the child when breastfeeding. In the case when it uses the expression "the nutritional needs of a mammal satisfied", we mean that during the greater part of the period of time from birth until the present age of the mammal's nutritional needs mammal fully satisfied. The term "diet to ensure the delivery of sialic acid in a quantity that is lower than the number, which normally enters the body of the child when breastfeeding, is CDATA, involving the introduction of a liquid composition in which the amount of sialic acid below 100 mg/L. According to other variants of implementation of the present invention, the number of sialic acid in the specified liquid composition is less than 200 mg/L.

According to the present invention, the composition that contains glycomacropeptide casein, may be administered to a mammal by any method enteral administration. Used herein, the term "oral introduction" means the administration of the composition in any Department of the gastrointestinal tract of a mammal and includes, without limiting the above, oral administration and enteral probe introduction.

Because glycomacropeptide casein may be administered to a mammal by itself, or together with other compounds or materials, it is a useful source of CGMP as one of the ingredients of the composition. The composition, which includes glycomacropeptide casein, can be any nutritional composition, preferably milk mixture. According to some variants of implementation of the present invention, the formula is a complete nutritional formula containing carbohydrates, fats and proteins. The number of lipids or fats can vary from 3 to 7 g per 100 kcal. The number of proteins typically in Rerum from 1 to 5 g per 100 kcal. The amount of carbohydrate usually varies from 6 to 16 g per 100 kcal.

Used herein, the term "composition" refers to man-made composition and, for example, does not apply to breast milk.

Sources of protein can be any known sources, such as skim milk, whey, casein, soy protein, hydrolyzed protein, amino acids and other Sources of carbohydrates can be any known sources, for example, lactose, glucose, corn syrup, maltodextrins, sucrose, starch, rice syrup and other Sources of lipids can be any known sources, including vegetable oils such as palm oil, soybean oil, palmolein, coconut oil, triglycerides of medium-chain, high oleic sunflower oil, high oleic safflower oil, etc.

As a basic framework for the introduction of CGMP can be used in some commercially available infant formula. For example, milk formula Enfamil® Lipil with iron (manufactured by Mead Johnson &Company, Evansville, Indiana, U.S.A) can be enriched with an effective amount of CGMP and used in accordance with the method stated in accordance with the present invention. Certain formulas, suitable for use in accordance with the present invention, described in the Examples below.

The total is the amount of protein in the composition, from all available sources is adequate for children and is usually from 12 grams per liter to 18 grams per liter and, according to some variants of implementation of the present invention, may be approximately 14 g per liter. The total number of sialic acid in the composition may range from 200 to 1500 mg per litre. Preferably, the claimed formula contains a liquid with a concentration of sialic acids, at least 200 mg per liter. According to some variants of implementation of the present invention, more preferably, the composition contains a liquid with a concentration of sialic acid, at least 300 mg per liter, more preferably 600 mg per litre. Preferably, the composition contains up to 6 g glycomacropeptide casein (CGMP) per liter or related protein fraction, commercially available from different sources and containing approximately 81% of protein and about 40 to 60 SA per gram protein; thus, the total number of sialic acid in the composition ranges from 194 to 1458 mg / l; or up to 6 g per liter fractions CGMP containing an increased amount of sialic acid.

The casein of glycomacropeptide, which is useful in accordance with the present invention, in General, can be isolated from any source and have any degree of purification or quality, suitable for use in the project for nutritional purposes or for inclusion in infant formula. Glycomacropeptide casein can be isolated from milk using any suitable method. For example, glycomacropeptide casein can be extracted from the concentrate obtained after the concentration of whey proteins. For example, this can be done by partial removal of lactose from concentrate followed by the addition of ethanol to cause precipitation. The supernatant is then collected and dried to obtain glycomacropeptide casein. A more detailed description of this process is presented in U.S. patent No. 5,216,129, which is included in the present description by reference and whose content does not contradict the information presented here. CGMP, useful for use in accordance with the method, as claimed in the present invention can also be obtained using the methods described in U.S. patents Nos. 6,555,659, 5,280,107, 5,968,586 and 5,075,424 as well as in PCT/ and 894/15952 and WO 03/049547. Alternatively, CGMP can be obtained from commercial sources, such as, for example, The Tatua Co-Operative Dairy Company Limited, Tatuanui, Morrinsville, New Zealand, MD Foods Ingredients amba of DK-6920 Videbaek, Denmark, DMV International of NCB-laan 80, NL-5460 BA Veghel, The Netherlands.

In accordance with the claimed method, preferably, glycomacropeptide casein or composition containing glycomacropeptide casein, is introduced in a quantity sufficient to ensure that iloveu acid in the body of a mammal at a concentration of 100 mg/kg / day, and, according to some variants of implementation of the present invention, the concentration of sialic acid 200 mg/kg / day is preferred.

According to one variant of implementation of the present invention, preferably, the total amount of protein in the composition is from 12 to 16 grams per liter, less than 40% by weight of the specified protein is glycomacropeptide casein. More preferably, the total amount of protein in the composition is from 13 to 15 grams per liter, with not more than 30% by weight of the specified protein is glycomacropeptide casein. According to one variant of implementation of the present invention, the protein composition represented by glycomacropeptides casinos and soy protein.

According to some variants of implementation of the present invention, uses new CGMP products in which the level of sialic acid is higher than in the standard CGMP products that are commercially available. These new products can be used alone or in combination in order to increase the content of sialic acid to a level close to the level in breast milk, taking into consideration the number of sialic acid in different ingredients. According to one variant of implementation of the present invention, glycomacropeptide casein includes glycomacropeptide the casein with a high content of sialic acid.

Used herein, the term "CGMP with high content of sialic acid" refers to containing glycomacropeptide casein (CGMP) fraction of milk that has been specially treated to increase the level of sialic acid and in which the level of sialic acid is higher than before treatment. CGMP products containing elevated levels of sialic acid, as described below in Examples 2 and 3.

One of these products, the approximate composition of which is described below in Example 2, hereinafter referred to as "CGMP with elevated levels of sialic acid" or "CGMP with high levels of sialic acid. Preferably, the amount of sialic acid CGMP in the specified product is more than 60 mg per gram of protein. Preferably, the amount of sialic acid exceeds 100 mg per gram of protein, more preferably, greater than 150 mg per gram of protein, even more preferably, 200 mg per gram of protein. Usually, the total protein content in the specified product is 50-60% by weight of the dry powder product, the content of sialic acid is about 190-230 mg per gram of protein, or approximately 100-130 mg per gram of powder. For comparison, a standard dry powder CGMP (for example, glycomacropeptide company Tatua Co-Operative Dairy Company Limited) contains 81% protein by weight, and the number of sialic acid is approximately 52 mg per gram of protein and 42 mg per gram of powder. It is obvious that the number of sialic acid in the specified product CGMP with high content of sialic acid increased 3 times (based on the weight of the powder) or 4 times (based on protein) compared to the standard CGMP products. For comparison, elektrodializnye (ED) whey powder contains 14% protein on a dry basis and approximately 30 mg of sialic acid per gram of protein, or 4.3 mg of sialic acid per gram of powder.

The advantage of using CGMP with high content of sialic acid as a source of protein in milk formulas lies in the fact that the number of sialic acid in the composition can be increased without replacing or reducing the number of standard sources of protein that are used in the composition. This feature is useful because it provides minimal disruption of the amino acid profile of protein composition.

According to a particular variant of implementation of the present invention, the level of the amino acid threonine in CGMP product with a high content of sialic acid is lower than in the source glycomacropeptide of a new product. Specified product type CGMP with high content of sialic acid in this description can also be indicated by the terms "CGMP with elevated levels of sialic acid and decreased levels of threonine or GMP with a high content of sialic acid and low content of threonine". The approximate composition of the product specified below in Example 3.

The number of sialic acid in the product CGMP with high content of sialic acid and low content of threonine is approximately 60 mg per gram of protein, and the concentration of threonine - less than 15 grams per 16 g of nitrogen. Preferably, the content of sialic acid is 100 mg per gram of protein, more preferably, is approximately 150 mg per gram of protein and, more preferably, 200 mg per gram of protein. Usually the content of sialic acid CGMP product with a high content of sialic acid and low content of threonine is from 85 to 150 mg of sialic acid (SA) per gram of powder, preferably, from 90 to about 140 mg SA per gram of powder, which is comparable with the content of sialic acid CGMP product with a high content of sialic acid. However, the content of threonine in CGMP product with a high level of sialic acid is one-fourth from the contents of the specified amino acids in commercial CGMP products. Preferably, the content of threonine in the product is less than 10 g per 16 g of nitrogen, more preferably less than 7 g per 16 g of nitrogen, more preferably less than 5 g per 16 g of nitrogen, and, even more preferably less than 4 g per 16 g of nitrogen. Alternative content threonine can be expressed another SPO is obom by weight calculated on the total weight of amino acids in a protein in this case, the content of threonine below 8% by weight calculated on the total weight of amino acids, preferably below 6%, even more preferably below 4%, even more preferably below 3%.

The advantage of the specified enriched in sialic acid product is that in addition to the high content of sialic acid without disrupting amino acid profile, as discussed above, can be controlled by the amount of threonine in the protein sources included in this formula. This can be useful in some embodiments to implement the present invention, when it is required to reduce the likelihood of hyperreninemia or another disease that is caused or potentiated by high levels of threonine in the diet.

For example, milk formula, which is useful in accordance with the present invention, may be made so that the content of sialic acid was at least 200 mg per liter, and the total protein content ranged from 12 to 16 grams per liter, less than 40% by weight is provided CGMP with high concentration of sialic acid. Preferably, the specified formula has a total protein content of 13 to 15 grams per liter, with not more than 30% by weight is provided CGMP with high concentration of sialic acid, more preferably mo is full-time the mixture has a total protein content of 13 to 15 grams per liter, not more than 15% by weight of CGMP is provided with a high content of sialic acid.

As an example, milk formula, stated in accordance with the present invention, may be made so that the content of sialic acid was at least 400 mg per liter, and the total protein content was from 13 to 15 grams per liter, with no more than 15% by weight of CGMP is provided with a high content of sialic acid.

Enriched with glycomacropeptide milk, notified in accordance with the present invention can be used in the same way as any other commercially available infant formula. It can be represented by a powder for subsequent breeding or liquid form. The composition should be Packed, stored and transported in the same manner as any similar product, and, in General, be used in the same way.

The following examples describe certain embodiments of the present invention. Other embodiments of the present invention, within the following claims, will be obvious to experts in this field. Assume that the specific embodiments of the present invention, and examples of their implementation are presented solely for the purpose of the illustration and better understanding of the present invention and are within the following claims of the invention without limiting its scope and nature. In all examples presented here, if not stated otherwise, all data are expressed in percent by weight.

REFERENCE EXAMPLE 1

This example describes the nutrient composition of commercially available milk mixture, suitable for enrichment of sialic acid, in accordance with the present invention.

Table 1
The nutrient composition of infant formula (Enfamil® Lipil with iron)
Nutrients (normal dilution)100 calories (5 FL oz)
Proteins, g2,1
Fat, g5,3
Carbohydrates, g10,9
Water, g134
Linoleic acid mg860
Vitamins
A, ME300
D, ME60
E, IU2
K, mcg 8
Thiamine (vitamin B1), mcg80
Riboflavin (vitamin B2), mcg140
B6, mcg60
B12, mcg0,3
Niacin, mcg1000
Folic acid, mcg16
Pantothenic acid, mcg500
Biotin, mcg3
C (Ascorbic acid)12
Choline, mg12
Inositol, mg6
Minerals
Calcium, mg78
Phosphorus mg53
Magnesium, mg8
Iron mg1,8
Zinc mg1
Manganese, mcg15
Copper, mcg75
Iodine, mcg10
Selenium, mcg2,8
Sodium, mcg27
Potassium, mg108
Chlorine mg63

The specified formula includes the following ingredients: whey with low mineral content, skim milk, vegetable oil (palm, soy, coconut and high oleic sunflower oils), lactose and a number of ingredients containing less than 1%: the oil fungus mortierella alpine, oil, Crypthecodinium cohnii, vitamin a palmitate, vitamin D3, acetate vitamin E, vitamin K1, thiamine hydrochloride, hydrochloride vitamin B6, vitamin B12, Niacinamide, folic acid, calcium Pantothenate, Biotin, sodium ascorbate, Inositol, calcium chloride, calcium phosphate, ferrous sulfate, zinc sulfate, manganese sulfate, copper sulfate, sodium chloride, sodium citrate, potassium hydroxide, sodium Selenite, taurine, nucleotides (adenosine 5'-monophosphate, citizen 5'-monophosphate, disodium salt guanosine 5'-monophosphate, the disodium salt of uridine 5'-monophosphate).

In accordance with the method, as claimed in the present invention, to this is oppozitsii you want to add, for example, glycomacropeptide casein in an amount sufficient to provide a concentration of sialic acid in the mixture, described in Table 1, 250 mg / l to about 1500 mg per litre. Added the number of sialic acid will be a portion of the total quantity of protein in the mixture (the total protein content of about 2.1 grams per 100 calories).

EXAMPLE 1

This example describes certain combinations of protein sources with a total content of sialic acid approximately 250 mg per liter. The ingredients shown in Table 2, can be used to replace the protein component kompoziziii described in Table 1.

Table 2
Sources of protein compositions And
Ingredientmg SA/g proteinand% protein ingredientGrams of ingredient per literGrams of protein per litermg SA/liter
The concentrate of whey proteins23,0035,0020,267,09163,08
6,3734,0015,385,2333,31
CGMPb52,0081,001,451,1761,07
Notes: a. "SA" in the table indicates sialic acid
b. CGMP refers to glycomacropeptide casein

EXAMPLE 2

This example describes certain combinations of protein sources with a total content of sialic acid to approximately 300 mg / l. The ingredients shown in Table 3, can be used to replace the protein component kompoziziii described in Table 1.

Table 3
Sources of protein composition In
Ingredientmg SA/g proteinand% protein ingredientGrams of ingredient per literGrams of protein per litermg SA/liter
Concentrate proteins23,0035,0037,0012,95297,85
serum
CGMPb52,0081,001,451,1761,07
Notes: a. "SA" in the table indicates sialic acid
b. CGMP refers to glycomacropeptide casein

EXAMPLE 3

This example describes certain combinations of protein sources with a total content of sialic acid to approximately 600 mg / l. The ingredients shown in Table 4, can be used to replace the protein component of the composition described in Table 1.

Table 4
Sources of protein composition In
Ingredientmg SA/g proteinand% Belk is in ingredient Grams of ingredient per literGrams of protein per litermg SA /liter
The concentrate of whey proteins23,0035,0013,004,55104,65
CGMPb52,0081,0012,009,72505,44
Notes: A. "SA" in the table indicates sialic acid
b. CGMP refers to glycomacropeptide casein

EXAMPLE 4

Table 5 presents the estimated complete nutrient composition of infant formula containing a total sialic acid approximately 250 mg per liter.

Table 5
The example formula, containing sialic acid
IngredientWeightNumber per 10,000 litres
Lactose (95% solids) 573,000 kg
Fat blend332,500 kg
The concentrate of whey proteins (36% protein, 5.8% of Ash)202,578 kg
Non-fat dry milk (36% protein, 52% carbohydrate)153,844 kg
Glycomacropeptide casein (CGMP, 81,18% protein)14,500 kg
Mono - and diglycerides7,233 kg
Calcium phosphate, rejonowy6,520 kg
Single-cell oil arachidonic acid6,485 kg
A dry mixture of vitamins for liquid compositions Enfamil AR5,250 kg
Ascorbic acid2924,250 g
Inositol834,750 g
Solid Zernovo the syrup 654,938 g
Taurine582,750 g
Niacinamide119,438 g
Calcium Pantothenate44,730 g
Vitamin b 12, 0.1% in the starch29,400 g
Biotin, 1% homogeneous25,095 g
Hydrochloride thiamine13,913 g
Riboflavin10,238 g
Hydrochloride pyridoxine8,138 g
Folic acid2,363 g
Concentrate lecithin3,694 kg
Potassium citrate3,350 kg
Unicellular wt is about docosahexaenoic acid amounting to 3.243 kg
A mixture of nucleotides powder Enfamil2,900 kg
Maltodextrin, 15 DE2552,290 g
Free from citizen 5-monophosphate acid202,710 g
The disodium salt of uridine 5-monophosphate59,740 g
Adenosine 5-monophosphate, free acid47,357 g
The disodium salt of guanosine 5-monophosphate37,903 g
Carragenan2,826 kg
Magnesium chloride1,657 kg
Calcium chloride, dihydrate1,200 kg
Chloride choline0,700 kg
Ferrous sulfate heptahydrate is 0,682 kg
Sodium citrate, dihydrate, granulated0,455 kg
A mixture of minerals in trace quantities w/Selenite powder0,392 kg
Zinc sulfate, monohydrate276,238 g
Powder of sodium Selenite 0,5%65,907 g
Copper sulphate powder29,510 g
Lactose, powder And16,323 g
Manganese sulfate,4,022 g
monohydrate
The mixture of vitamins A, D, E, K in the liquid mixture Enfamil0,324 kg
Acetate tocopherol160,882 g
Soybean oil 139,612 g
Palmitate vitamin a17,253 g
Concentrate cholecalciferol5,715 g
Vitamin K liquid0,538 g
Ascorbic acid0,150 kg
L-carnitine0,150 kg
Detonirovanie water q.s.10310,986 kg
Hydrochloride potassium---

In Tables 6 and 7 presents the content of specific components in the composition described in Table 5 percentages: 1) mass mass (2) mass to volume, and 3) calories. Specific density specified a particular composition is 1.0310986.

Table 6
Composition formula
Component% weight/weight % weight/vol.
Proteins1.381.42
Fats3.503.61
Carbohydrates7.207.43
Balance0.370.38
The total solids content12.4512.84

Table 7
The distribution of calories in the milk mixture
Component%
Proteins8.38
Fats47.83
Carbohydrates43.79

EXAMPLE 5

Table 8 presents the nutritional composition described in Example 4, 100 calories, and 100 milliliters of composition.

63
Table 8
The nutrient composition of the milk mixture
At 100 calories100 ml
Calories, calories10068
Proteins, g2.11.42
Fat, g5.33.6
Carbohydrates, g10.97.4
Linoleic acid mg860580
Linolenic acid mg8054
Arachidonic acid mg3423
Docosahexaenoic acid mg1711.5
Vitamin a, ME300200
Vitamin D, ME6041
Vitamin E, ME21.35
Vitamin K1, mcg128.1
Thiamine, mcg12081
Riboflavin, mcg14095
Vitamin WB, mcg6041
The vitamin B12, mcg0.50.3
Niacin, mcg1200812
Folic acid, mcg1610.8
Pantothenic acid, mcg500340
Biotin, mcg32
Vitamin C mg149.5
Choline, mg128.1
Inositol, mg64.1
Taurine, mg64.1
L-carnitine, mg21.35
Calcium, mg7853
Phosphorus mg5336
Magnesium, mg85.4
Iron mg1.81.2
Zinc mg10.68
Manganese, mg2617.6
Copper mg8557
Iodine mg1510
Sodium, mg2718.3
Potassium, mg10873
Chlorine mg43
Selenium mg2.81.89
Sialic acid mg3725
The ratio of calcium/phosphorus---------
AMP Equivalents, mg (a)0.50.34
IDA Equivalents, mg (a)2.51.69
GMP Equivalents, mg (a)0.30.20
UMP Equivalents, mg (a)0.90.61
Equivalents of nucleotides, mg (a)4.22.84
TPAN-AMP mg----
TPAN-CMP, mg----
TPAN-GMP mg----
TPAN-UMP mg----
The total content of TPAN----
TPAN-CMP/TPAN-GMP----
ratio
Note: the Total content of nucleotides and the corresponding nucleosides, expressed by weight of nucleotides.

REFERENCE EXAMPLE 2

This example illustrates the creation of CGMP product with a high content of sialic acid.

The fraction of cheese whey, enriched with GMP, fractionary using anion chromatography to obtain fractions enriched in sialic acid. The amino acid profile of the product is similar amino acid profile of commercially available GMP (for example, the company Tatua Co-Operative Dairy Company Limited, Tatuanui, Morrinsville, New Zealand), the content of sialic acid in 1.5-3 times higher than in the standard product.

From the obtained fractions enriched in sialic acid, can be removed all salt by electrodialysis, if necessary, after which the fraction is dried for having Holocene dry powder, which can then be used to add liquid or powder formula. This product is CGMP product with a high content of sialic acid and, at the time of filing, as supplied by Tatua Co-Operative Dairy Company Limited; the products are referred to as H, H, H and H. The protein, sialic acid, amino acid profile of these products are presented in Table 9.

REFERENCE EXAMPLE 3

This example illustrates the receipt of CGMP product with a high content of sialic acid and low content of threonine.

The fraction of cheese whey, enriched with GMP, is subjected to partial proteolytic hydrolysis with subsequent fractionation using anion chromatography to obtain fractions enriched in sialic acid with a low content of threonine. The content of sialic acid in the specified product in 1.5-3 times higher than its content in commercially available CGMP products, but the level of threonine is one-fourth of the level in the original CGMP material.

From the obtained fractions enriched in sialic acid with a low content of threonine, can be removed all salt by electrodialysis, if necessary, after which the fraction is dried to obtain a dry powder, which you can then use the diamonds to add in liquid or powder formula. This product is CGMP product with a high content of sialic acid and low content of threonine to the time of the application supplied by the company Tatua Co-Operative Dairy Company Limited; the product is denoted as W 4733. The protein, sialic acid, amino acid profile of these products is presented in Table 10.

Table 10
Amino acid profile and content of sialic acid four CGMP products with a high content of sialic acid and low content of threonine
Amino acidCGMP with high content of sialic acid and low content of threonine
W4733CGMP powderED powder whey
Arginine2.31.4
Histidine-1
Isoleucine13.1 11.6
Leucine5.34.6
Lysine3.28.3
Methionine0.71.5
Cystine0.10.2
Phenylalanine01.8
Tyrosine00.4
Threonine3.815.9
Tryptophan00
Valine16.39.7
Alanine15.96.6
Aspartic acid6.311.1
Glutamic acid38.926.2
Glycine2.51.5
Proline16.914.2
Serine08.1
Total125.4124.1
% protein8114.31
mg SA/g protein5229.92
mg SA/g38.03 42.124.28
powder
(re-analysis)117.02
Average127.525
The number of amino acids are expressed in grams of amino acids per 16 grams of nitrogen
CGMP powder is commercially available glycomacropeptides supplied by Tatua Co-Operative Dairy Company Limited
ED whey powder is a commercially available electrodialysing whey powder
Samples W4731, W4733 and W4735 represent CGMP with high content of sialic acid and low content of threonine and available in the company Tatua Co-operative Dairy Company Limited, Tatuanui, Morrinsville, New Zealand.

As mentioned above, the level of threonine in the new product is one-quarter of its level in a commercial product. Accordingly, it is believed that the use of CGMP enriched in sialic KIS is Auteuil, with low levels of threonine in formula allows you to create a composition with a high level of sialic acid and normal, the desired protein content of about 14 g of protein per liter, and the desired amino acid profile with low levels of threonine.

EXAMPLE 6

This example describes the use of CGMP fraction enriched in sialic acid in milk mixture.

CGMP product enriched in sialic acid, can be used as a source of protein in the milk mixture in the same way as the powder serum or normal CGMP powder. Table 11 shows the expected number of sialic acid in the milk mixture, the protein composition which is provided by traditional sources. The number of CGMP powder is strictly limited in order to avoid unwanted deviations amino acid profile of proteins from the desired standard profile for the dairy mixture.

Table 11
The content of sialic acid in the milk mixture, the protein composition which is provided from traditional sources
Source of proteinSialic acid, mg/ protein gPR is cent protein Protein, g/lSialic acid, mg/l
The concentrate of whey proteins2335%6.82156.77
Non-fat dry milk slightly warmed6.3734%6.2539.8
CGMP product enriched in sialic acid21454.93%1.11237.54
Total14.17434.12

CGMP powder used in this composition may be replaced by a new CGMP product enriched in sialic acid, as described above in reference example 3. Table 12 shows that in this case, the total content of sialic acid increases more than two times without breaking the amino acid profile of the protein.

Table 12
The content of sialic acid in milk mixture, Belkova the composition of which is provided by traditional sources, and the new CGMP product enriched in sialic acid
Source of proteinSialic acid, mg/protein gThe percentage of proteinProtein, g/lSialic acid, mg/l
Concentrate proteins2335%6.82156.77
serum
Skimmed milk powder Low heat6.3734%6.2539.8
CGMP product enriched in sialic acid21454.93%1.11237.54
Total14.17434.12

In case CGMP powder enriched in sialic acid, is used in the dual number, as described above, when the consumption of skimmed milk powder with the content of sialic acid in milk mixture may rise to the level presented in Table 13.

Table 13
The content of sialic acid in the milk mixture, the protein composition which is provided by traditional sources, as well as new CGMP product enriched in sialic acid
Source of proteinSialic acid, mg/protein gThe percentage of proteinProtein, g/lSialic acid, mg/l
The concentrate of whey proteins2335%6.82156.77
Skimmed milk powder Low heat6.3734%5.1432.74
CGMP product enriched in sialic acid21454.93%2.22475.08
Total14.18664.59

EXAMPLE 7

what this example illustrates the effectiveness of diet, including glycomacropeptide casein, in the improvement of processes of learning and memory in pigs.

Methods

Animals

Within 20 months from the industrial pigsty take home male piglets at the age of 3 days (Sus scrofa) weighing from 1.5 to 2.4 kg from 14 different litters. Animals distributed to weight and randomly divided into 4 groups for different treatment. Pigs contain pairs, according to the chosen diet, in wire pens with a concrete floor in a room with controlled temperature. Each bathroom has described "the nest" (rubber tire, covered with a towel), lamp heating and identical plastic toys. Pigs are trained to use the far corner of the paddock as toilets, stalls cleaned daily. To organize the next light cycle: 12 h light (8 am - 8 pm)/dark (8 pm - 8 am).

CGMP support

Glycomacropeptide casein (CGMP) content of sialic acid 60 mg/g provided by the company Tatua Dairy Cooperative (Morrinsville, New Zealand) and is added to the substitute pork milk in the specific quantity Woombaroo Food Products (Glen Osmond, Australia). The number of sialic acid in the final formula varies depending on the group: 77 mg/l (group 1, control group receives the mixture without adding CGMP), 250 mg/l (group 2, low dose), 600 mg/l (group 3, the average dose) and 842 mg/l (group 4 high dose). Shown is the concentration qualifies for 25, 71, 171 and 240 mg of sialic acid per kg of body weight. Substitutes are such that the total protein intake remained the same, regardless of the number of added CGMP. To maintain the normal growth of the piglets received 285 ml of milk per kg of body weight in the first 2 weeks of the study and 230 ml per kg of body weight during the remaining weeks. Feeding is carried out in 8:00, 13:00, 18:00 and 23:00, advanced night give 50 ml of milk. Body weight, consumption of milk and medicines are registering daily.

Assessment of learning ability and memory

Formal testing of the ability to learn to start with a 23-day-old piglets by means of the maze with the radial location of the 8 compartments (figure 1 and figure 2). The labyrinth is a specially designed wooden structure located in the educational sector (4.2 m × 4.4 m), which is connected with the home paddock. In order to reduce stress, couples piglets allowed to explore the maze the day before the formal study. Perform two tests on learning: task 1 and task 2. For each job in one compartment of the maze is placed available milk, and in the remaining 7 - unavailable milk, so that all 8 compartments of the maze comes the same olfactory signal. For each job on the door of the compartment containing the available m the Loco, put a visual signal consisting of 3 points (FIGURE 3). In the first task (easy job) one black dot is placed on the door of the other seven compartments. In the second task (more complicated) for the remaining 7 doors put two black dots.

All pigs tested in the maze individually. Forty experiments task 1 carry for 5 days (8 experiments in the day) and 40 experiments task 2 is carried out for 6 days. Assessing the speed of learning is determined by counting the number of repetitions needed to successfully learn a visual signal, and the number of errors (wrong door) and successful attempts (right door) to find the available milk in each experiment. The error is considered each case, when the pig is in the wrong door or just pokes his head through the wrong door. Successful execution of the job is when the pig enters through the right door and finds the available milk. The criterion for learning the visual signal is a maximum of one error in three consecutive experiments. Two days after the completion of experiments on task 1 and 2 perform similar tasks or "memory test", but for each job perform only one experiment. All tests evaluated 2 groups of researchers, aware of to whom icesto consumed sialic acid, however, the results of tests later confirmed by an independent analysis of the video.

Evaluation of stress

Because stress can affect learning or memory, for measuring the level of cortisol in the blood in the morning, with weekly intervals, starting from the 7th day using a commercially available kit (Coat-A-Count Cortisol, Diagnostic Products, Doncaster, Australia). Variations between the experiments and the experiments were 9% and 12%, respectively.

Analysis of the quantity associated with gangliosides and proteins sialic acid

34 and 35 day of pigs killed by injection of sodium pentothal (50 mg/kg). The brain tissue from the frontal cortex is collected and stored at -80° C until analysis. The number of sialic acid associated with gangliosides and proteins, determined separately using known literature methods (see Mahal, Id). All samples analyzed twice the final concentration of sialic acid in each fraction is expressed as µg/g of fresh tissue.

Statistical analysis

The difference in speed of learning (number of experiments, which is required for learning visual signal) is compared using analysis of durability of the Kaplan-Meier Cox regression to explore possible covariances, which can affect the speed of learning. Comparison of averages (without covariances and with them) OS the effect using the General linear model (univariate ANOVA). The Pearson correlation is used to examine the relationship between the number of errors, successful attempts, body weight and performance memory. All statistical analysis completed using SPSS for Windows 11 Inc, Chicago. Set the confidence level of 0.05.

Results

Speed training

As in the easy and difficult task, groups of pigs fed a diet enriched in sialic acid, was studied visual signal faster than piglets from the control group (Kaplan-Meier, P=0.0014 in task 1 and P=0.0177 in task 2, see FIGURE 4 and FIGURE 5). In task 1, only 45% of the piglets from the control group achieved results in 40 experiments, while the best indicators of training were recorded for group 2 (100% piglets learned signal 40 experiments), then for a group of 4 (80%) and group 3 (70%). In task 2 was identified relationship between dose and response (P=0.0177), while pigs from all 4 groups had achieved results of 30 experiments. Differences between groups remained significant when studying the differences of body weight during the study (P<0.05).

Covariance learning

Apparently, the pigs used the information obtained during task 1, for storing a visual alert in task 2. Therefore, the researchers used the total number of errors and successful attempts in task 1 as to the variations for the analysis of learning rates in task 2. Differences between groups after the amendment was relatively high (P=0.002 errors as covariances & 0.004 successful attempts as covariances), but dose-dependent effect of speed training in the task 2 did not change. The results were considered similar, if taken into account all 40 experiments in task 1, or only the last 20 experiments. The number of errors in experiments 21-40 task 1 also varied significantly between groups. This was the case with or without the use of errors in experiments 1-10 as covariances (P=0.016). The control group made a significant greater number of errors than group 2 (P=0.005) and group 4 (P=0.006), but not significantly more than group 3 (P=0.06). In a similar analysis for task 2, the covariance was significant, but groups did not differ significantly (covariance or not) (P>0.05) (FIG.6). A similar analysis using the total number of errors in all experiments revealed significant differences between experimental groups (combined groups 2, 3 and 4) and control group (P=0.009) and minor differences between groups in task 2 (p=0.048).

Test memory

In piglets, which were able to pass a test on training, learning the ability to remember visual signal within 48 hours. The overall differences between the groups was the way the almost significant for task 2 (P=0.036), but not for task 1 (P=0.165, FIG.7 (A)).

Dose-dependent effect was not observed in any of the jobs. In the 2nd task, the performance of the group 3 and control group were approximately equivalent (FIG.7 (C)). When combining the results of groups 2, 3 and 4 (experimental groups) number of errors was less than 35% (mean=1.5) compared with the control group (mean=2.4, p=0.036).

It is not surprising that the number of errors and successful test runs during the study period significantly affects the process of learning all the pigs. Thus, if during the period of study was made of a large number of errors, we can assume that in the test for the memory will also be a lot of errors (P=0.03 in task 1 and P=0.029 in task 2, the bilateral correlation of Spearman). Similarly, a large number of correctly performed experiments during the first job is associated with a small number of errors in the test on the memory in task 1 (P=0.007), but not in task 2 (P=0.20). Body weight, weight gain and speed training had no significant effect on the test for memory (P>0.05).

The level of cortisol in plasma

The average concentration of cortisol in each group during the 5 weeks of the study presented in FIG (A). In all groups, differences in the concentration in the first week and during the remaining weeks were significant (weeks the La 1 vs 2, 3, 4 and 5 weeks; P=0.031, 0.015, 0.001 and 0.001 respectively). Since 2 weeks and then the concentration of cortisol in serum from all pigs were on average lower, but not significantly (P>0.05). When all experimental groups were combined into one group, the differences between the experimental groups and the control group were insignificant (P>0.05, FIG ()). On the 2nd week of differences between group 2 and group 3 were significant (P=0.044). In the case when the concentration of cortisol is used as the covariance during training, significant differences are not observed (P>0.05).

The concentration of sialic acid in the brain

A large part of sialic acid in the frontal cortex of pigs (86%) was associated with gangliosides, a small fraction (~13%) was associated with a glycoprotein, an even smaller proportion (2%) were in the free form. These data are consistent with previously published data Brunngraber, E.G. et al., Brain Res., 38:151-162 (1972) and Mahal, L.K. et al., in J. Biolo. Chem., 277:9255-9261 (2002). Was a significant dose-dependent relationship between content-related protein sialic acid in the brain and the amount of CGMP in the diet, for example, for a group of 4 experienced the highest level, then there were a group of 3, 2 and 1. Level in group 1 was significantly lower than in group 4 (P=0.001) and group 3 (P=0.001), but not in group 2 (P=0.126). On average, sod the neigh-related protein sialic acid in the gray matter was 6-10% higher in groups, receiving CGMP than in the control group (P=0.000, table 14).

Table 14
The average level of sialic acid associated with gangliosides, sialic acid-related protein, and free sialic acid in the frontal cortex, depending on the amount of typing CGMP
GroupnaAssociated with ganglioside SA (µg/g)Associated with protein SA (µg/g)Free SA (µg/g)Total (ug/g)
averageSEaverageSEaverageSEaverageSE
Group 114162911423.00.227910
Group 213176912123.10.230010
Group 314182912623.00.231110
Group 4121851012723.20.231510
P value : 0.3070.0010.8760.068
Notes: a. "n" is the number of piglets in the group

The average concentration of sialic acid associated with gangliosides, was also dose dependent (13%, 11% and 8% higher in the group 4, group 3 and groups is 2, respectively, vs control), but the differences were not statistically significant (P=0.089, 0.124 and 0.299, respectively, table 14). When all groups treated with CGMP, combined into one group, the differences between the experimental groups and the control group was insignificant (P=0.07, FIGURE 9). Variations between individual indicators of sialic acid associated with gangliosides, were higher than the variation of sialic acid-related proteins (CV=~5% for related gangliosides vs 2% for the related protein sialic acid). The same phenomenon was obtained in the study of the brain of children rights, held Mahal, L.K., Id.

The total concentration of sialic acid (associated with gangliosides + - related proteins) was slightly higher in the combined experimental group treated with CGMP, than in the control group (P=0.051), but not the concentration of free sialic acid (R=0.58, FIGURE 9).

The correlation between the concentration of sialic acid in the brain and learning

High concentration of sialic acid associated with gangliosides, sialic acid-related proteins, and total sialic acid is associated with a more rapid learning in both tasks 1 and 2 (see FIGURE 10 (A), FIGURE 10 (B)11 (a) and 11 (B)).

None of the correlations, parametric or nonparametric, did not reach statistical significance (see FIGURE 10 and 11). Anallogichnom analysis test on the memory negative correlation (high content of sialic acid, a small number of errors) were found in both tasks 1 and 2, however, the results again did not reach statistical significance (P>0.05), except for the related protein sialic acid in group 4 for task 1 (P=0.045, FIG). There was a significant positive correlation between the total number of successful results and the concentration of sialic acid associated with gangliosides, in task 1 (P=0.045), but not in task 2 (P 0.05).

Gain weight

Average (±SE) of initial body weight was similar for each group (2.1±0.04 kg), all animals gained body weight with the same pace (FIG). Despite the fact that the control group weighed significantly more than other groups at the end of the study, the rate of weight gain (g / day) among groups were not significantly different (P=0.503).

All references cited herein, including publications, patents, patent applications, reports, reviews, manuscripts, theses, pamphlets, books, Internet sites, magazine articles, periodicals, etc. included in the description only as the quoted material. Discussion of links is intended to summarize the statements of the authors of these publications here and not made any assumptions about their occurrence in the prior art. The applicant leave the right to challenge the accuracy and validity of citire what's sources of information.

From the above description of the obvious benefits that can be achieved using the present invention, and the advantages of the obtained results.

Disclosed in the description and other modifications and variations of the present invention can be carried out by a specialist in this field of knowledge without going beyond the scope of the invention, which is reflected in the formula. In addition, it should be clear that various aspects and embodiments of the invention can be partially or completely modified without changing its essence. Specialist in this field is also clear that given in this application is the description of the invention serves only for illustrative purposes and in no way limits the scope of the invention, the essence of which is expressed in the formula. Thus, the content and breadth of the claims of the claims is not limited to the description that characterizes the preferred embodiment of the invention.

1. Method for improving learning and/or memory in the human child, which includes the introduction of baby formula, containing glycomacropeptide casein in an amount sufficient to ensure that the child's body, at least 100 mg/kg / day sialic acid

2. The method according to claim 1, in which the age of the child varies from 1 day to 4 years.

3. The method according to claim 1, in which the ω child is a child in the neonatal period of development.

4. The method according to claim 1, in which the need for improving learning and/or memory occurs when at least part of the nutritional needs of the child are met by the introduction of the milk mixture with the content of sialic acid of less than 100 mg per litre.

5. The method according to claim 4, in which a large part of the protein composition represented soy protein or proteins in cow's milk.

6. The method according to claim 5, in which at least 75% (by weight) protein composition represented soy protein or proteins in cow's milk.

7. The method according to claim 5, in which almost the entire protein composition represented soy protein or proteins in cow's milk.

8. The method according to claim 1, in which the formula is balanced in nutrients and contains proteins, fats and carbohydrates.

9. The method of claim 8, wherein the protein is selected from the group comprising the proteins of cow's milk, soy proteins and their mixture.

10. The method according to claim 1, in which the number of glycomacropeptide casein in the milk mixture is sufficient to ensure receipt in an organism of the child, at least 200 mg/kg / day of sialic acid.

11. The method according to claim 1, wherein the total protein content in the milk mixture ranges from 12 to 16 grams per liter, less than 40% by weight, it is provided by glycomacropeptides casein.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: present invention relates to bioorganic chemistry and medicine and can be used to prepare medicinal agents with neurotropic activity. The technical result is achieved due to creation of a peptide with neurotropic activity and having general formula: Ac-X-Met-Pro-Arg-Gly-NH2, where X-Met - L-Met or DL-Met.

EFFECT: obtaining a novel compound - analogues of the C-terminal end of vasopressin which are capable of selectively bonding with different subtypes of vasopressin receptors and, as a result, have different types of neurotropic activity, have low toxicity and are suitable for simple intranasal administration.

5 ex

FIELD: medicine.

SUBSTANCE: there is claimed application of dipeptidyl peptidase IV inhibitor (DPP-IV inhibitor), vildagliptin or its salt for production of medication for prevention, retardation of progress or treatment of peripheral diseases such as peripheral neuropatia, neurodegenerative disorders, cognitive disorders, as well as for improvement of memory and ability to learn, and pharmaceutical composition for the same purpose. It is demonstrated: vildagliptin increases stage of wakefulness and response to external stimuli, increases REM sleep phase.

EFFECT: combination of vildagliptin with donepezil considerably improves disturbed ability to learn.

23 cl, 5 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to a compound having the structural formula I , or its pharmaceutically acceptable salt, ester or amide, where A has the structure , where each bond in A, represented by a dotted and a solid line, represents a carbon-carbon single bond; each of a, b and c is a carbon atom; each of e, f, g and h is a carbon atom; X is nitrogen; X' is C; L is absent; each n equals 1; Y is nitrogen; W is nitrogen; R1 is hydrogen; each of R2, R3 and R4 is a hydrogen atom; each of R6, R8 and R9 is a hydrogen atom; R5 is selected from a group consisting of halogen, C1-6alkyl, optionally substituted with a hydroxy group, and C1-6alkoxy; R7 is selected from a group consisting of halogen, C1-6alkyl and perhalogenalkyl; Z is selected from a group consisting of NR11, oxygen and CH2; R11 is hydrogen; and each bond in formula I, represented by a dotted and a solid line, is a carbon-carbon double bond. The invention also relates to a method for synthesis of a formula V compound, a pharmaceutical composition based on a formula I compound, methods of treating psychoneurological disorders, a pharmaceutical composition containing a formula I compound and a psychoneurological agent.

EFFECT: obtaining novel compounds useful for modulating muscarine receptor activity.

37 cl, 1 tbl, 141 ex

FIELD: medicine.

SUBSTANCE: invention refers to psychoneurology, particularly to an agent for treating multiple sclerosis. A composition (solid or liquid dosage form) contains triiodide 1,3-diethylbenzimidazolium as an agent, low-molecular surgical polyvinylpyrrolidone, presented as a solubiliser, and an agent stabiliser, and in addition in the liquid dosage form - ethanol as a solvent.

EFFECT: composition under the invention exhibits high therapeutic effectiveness in treating multiple sclerosis, and is characterised by relieving undesirable by-effects.

2 cl, 4 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel substituted 2-alkylsulfanyl-3-arylsulfonyl-pyrazolo[1,5-a]pyrimidines of general formula 1, their pharmaceutically acceptable salts and/or hydrates having serotonin 5-HT6 receptor antagonist properties. In general formula 1 , R1 and R3 independently denote optionally identical C1-C3 alkyl, and R2 is a -(CH2)nX group or R1 and R3 independently denote different substitutes selected from C1-C3 alkyl or a -(CH2)nX group, and R2 is a hydrogen atom or C1-C3 alkyl; R4 is C1-C3 alkyl; Ri5 is a hydrogen atom, one or two identical or different halogen atoms, C1-C3 alkyl; equals 0, 1 or 2; n equals 0, 1, 2 or 3; X is a carboxyl CO2H, C1-C3 alkyloxycarbonyl, aminocarbonyl CONR6R7 or amino group NR6R7; except compounds in which R3 is a -(CH2)nX group, where X is an amino group NR6R7 and n equals 0; R6 and R7 are optionally identical and denote a hydrogen atom, optionally substituted C1-C5 alkyl or R6 and R7 together with the nitrogen atom with which they are bonded form an optionally substituted 6-member azaheterocyclyl containing 1-2 nitrogen atoms in the ring, where the substitute is selected from C1-C3 alkyl.

EFFECT: obtaining compounds which can be used in treating diseases of the central nervous system during prevention or treatment of cognitive disorders, neurodegenerative diseases, psychiatric disorders, have anxiolytic and nootropic effect and can be used to prevent and treat anxiety disorders and enhance mental capacity.

25 cl, 2 tbl, 12 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel substituted 3-arylsulfonyl-pyrazolo[1,5-a]pyrimidines of general formula 1, their pharmaceutically acceptable salts and/or hydrates having serotonin 5-HT6 receptor antagonist properties. The compounds can be used in treating and preventing various diseases of the central nervous system, pathogenesis of which is associated with 5-HT6 receptors, particularly cognitive disorders, neurodegenerative diseases and psychiatric disorders. The compounds have anxiolytic and nootropic effect and can also be used for preventing and treating anxiety disorder and for enhancing metal capacity. In formula 1 , R1 is a hydrogen atom, C1-C3 alkyl, (C1-C3)alkyloxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, pyridyl; R2 is a hydrogen atom, halogen atom, C1-C3 alkyl, phenyloxy or pyridyloxy; R3 is a hydrogen atom, C1-C3 alkyl, C1-C3 alkyloxy, (C1-C3)alkyloxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, pyridyl; R4 is C1-C3 alkyl; R5 is a hydrogen atom, one or two halogen atoms, C1-C3 alkyl, C1-C3 alkyloxy or hydroxyl; X is a sulphur atom or thionyl group (SO).

EFFECT: obtaining compounds which can be used in treating and preventing various diseases of the central nervous system.

24 cl, 9 dwg, 4 tbl, 19 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel 2-alkylamino-3-arylsulfonyl-pyrazolo[1,5-a]pyrimidines of general formula 1, their pharmaceutically acceptable salts and/or hydrates having serotonin 5-HT6 receptor antagonist properties. The compounds can be used to treat and prevent development of various diseases of the central nervous system, pathogenesis of which is associated with 5-HT6 receptors, particularly Alzheimer's disease, Parkinson's disease, Huntington disease, schizophrenia, other neurodegenerative diseases and cognitive disorders. In general formula 1 , R1 is a hydrogen atom, C1-C3alkyl, C1-C3alkyloxyC1-C3alkyl, hydroxyC1-C3alkyl, adamantyl, optionally substituted phenyl, 5-6-member heterocyclyl containing a nitrogen or oxygen heteroatom, possibly condensed with a benzene ring; R2 is a hydrogen atom, halogen atom, C1-C3alkyl, pyridyloxy; R3 is a hydrogen atom; C1-C3alkyl; optionally substituted amino group selected from amino, mono- or di(C1-C3alkyl)amino, di(C1-C3alkyl)aminoC2-C3alkylamino, N-[di(C1-C3alkyl)aminoC2-C3alkyl]-N-(C1-C3alkyl)-amino; C1-C3alkyloxy, C1-C3alkyloxyC1-C3alkyl, hydroxyC1-C3alkyl; 6-member azaheterocyclyl, possibly containing an extra nitrogen or oxygen atom, a -N(C1-C3alkyl) group; R4 is C1-C3alkyl; R5 is a hydrogen atom, one or two halogen atoms or C1-C3alkyl.

EFFECT: design of a method of obtaining compounds, a pharmaceutical composition and a medicinal agent for treating and preventing various diseases of the central nervous system.

25 cl, 12 dwg, 3 tbl, 20 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine and aims at treating the conditions associated with amnesia and impaired cognition. A medically indigent patient is prescribed with the effective amount of a composition containing a mixed extract prepared of Scutellaria plants with high concentration of free V-ring flavonoids, including Baicalin, and an extract of Acacia plants with high concentration of flavanes, including catechine and epicatechin.

EFFECT: methods ensure restoration and preservation of cognition and memory.

44 cl, 15 ex, 2 tbl, 17 dwg

FIELD: chemistry.

SUBSTANCE: invention refers to novel compounds of the general formula (I) , where R1, R2 are independently H or C1-C6-alkyl; R3, R4 are independently H or C1-C6-alkyl; R5 is halogen, CN; n, m or o are 0, 1 or 2; and to pharmaceutically acceptable salts thereof.

EFFECT: compounds with monoaminooxidase B inhibition properties applicable in obtainment of pharmaceutical drugs with relevant effect.

14 cl, 3 dwg, 31 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to serotonin 5-HT6 receptor antagonists - new substituted 3-sulphonyl-6,7,8,9-tetrahydro-pyrazolo[1,5-a]pyrido[4,3-e]pyrimidines of formula and substituted 3-sulphonyl-5,6,7,8-tetrahydro-pyrazolo[1,5-a]pyrido[4,3-c1]pyrimidines of general formula 2, a medicinal base and pharmaceutical compositions containing the medicinal base in form of the said compounds, as well as to a method of treating and preventing development of different conditions and diseases of the central nervous system of humans and warm-blooded animals. In formulae and Ar is phenyl which is optionally substituted with halogen atoms, or a 6-member heteroaryl which contains a nitrogen atom in the ring; R1 is a hydrogen atom, C1-C3alkyl, hydroxy C1-C3alkyloxy group, C1-C3alkylsulphanyl group; R2 is a hydrogen atom or C1-C3alkyl, R3 is a hydrogen atom optionally substituted C1-C3alkyl or tert-butyloxycarbonyl.

EFFECT: obtaining compounds for preventing development of different conditions and diseases of the central nervous system of humans and warm-blooded animals.

16 cl, 3 tbl, 1 dwg, 10 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and biotechnology and concerns an antimicrobial peptide compound. Substance of the invention consists in use of an amino acid compound containing the sequence IGKEFKRIVERIKRFLRELVRPLR for preparing a drug for fungal or bacterial infection in a mammal. Besides said compound exhibits sensitivity to toxins, and especially to bacterial toxins, such as lipopolysaccharide or lipoteichoic acid.

EFFECT: extended field of use in comparison with antibiotics.

22 cl, 9 ex, 4 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to oncology, and concerns therapy of intestinal adenoma and/or adenocarcinoma by Notch inhibition. That is ensured by introduction of a γ-secretase inhibitor or a based pharmaceutical composition in the effective amount.

EFFECT: effective therapy enabled due to differentiation of adenoma and/or adenocarcinoma cells into goblet cells.

20 cl, 1 tbl, 6 dwg

FIELD: medicine.

SUBSTANCE: methods according to the invention consist in introducing an Aβ 16-23 fragment having an amino acid sequence KLVFFAED of 16-23 residues SEQ ID NO:1. Besides the invention concerns the Aβ 16-23 fragment and a pharmaceutical composition containing it.

EFFECT: feasible prevention and treatment of Alzheimer's disease ensured by inhibition of amyloid deposition in cerebrum.

72 cl, 2 dwg, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to glucagon-like peptide 1 secretion stimulation and to foodstuffs and beverages stimulating glucagon-like peptide 1 secretion and containing k-casein as the active component. declaration is made for a medicinal glucagon-like peptide-1 secretagogue containing k-casein as the active component.

EFFECT: invention ensures glucagon-like peptide 1 secretion stimulation and postprandial hyperglycemia inhibition.

2 cl, 2 ex, 4 tbl, 2 dwg

FIELD: medicine.

SUBSTANCE: treating cardiovascular diseases caused by fibroid heart is ensured by introducing compositions containing vasoactive intestinal peptide (VIP) or its active fragments, in the developed doses and regimen.

EFFECT: involution or prevented development of fibroid heart.

16 cl, 10 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: invention represents a combination containing VEGF Trap and 5-fluorouracil to be applied in treatment of neoplasms.

EFFECT: higher effectiveness of the combination ensured by therapeutic synergism of its components, and reduced toxicity.

4 cl, 1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to ophthalmology. It involves intradermal or open patch retinalamine test. A papule of diametre 5 mm and more shows ophthalmia.

EFFECT: method allows simplifying and improving effectiveness of diagnostics.

2 ex

FIELD: medicine.

SUBSTANCE: invention is related to nucleic acids and multidomain proteins, which are able to bind vessel endotheliocyte growth factor (VEGF), and may be used in medicine. Recombinant method is used to produce polypeptide, which consists of component (R1R2)X and, unnecessarily, multidomain component (MC), which represents aminoacid sequence with length from 1 to 200 of amino acids, having at least one remainder of cysteine, where X≥1, R1 means antibody-like (Ig) domain 2 of VEGF receptor Llt-1, and R2 means Ig-domain 3 of VEGF receptor Flk-1. Produced fused polypeptide does not contain multidomain component in case, when X=2, and in case when X=1, multidomain component represents aminoacid sequence with length from 1 to 15 amino acids. Produced polypeptide is used in composition of pharmaceutical compound for VEGF-mediated disease or condition.

EFFECT: invention makes it possible to produce highly efficient trap of VEGF, special structure of which is suitable for local introduction into specific organs, tissues or cells.

16 cl, 3 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: as a result of substitution of part of amino-acid sequence of human protein PRG-4 coded with exon 6 with artificial polypeptide, including 4-15 sequences KEPAPTT or KEPAPTT-like sequences, recombinant proteins are produced with considerably lower number of glycolised repetitions compared to natural lubricin, which at the same time preserve its lubricating properties. It is suggested to use new recombinant proteins in pharmaceutical compositions and in methods of treatment, where natural lubricin is traditionally used.

EFFECT: application of the present invention for medical purposes has certain advantages related to simplification, compared to native protein.

28 cl, 1 dwg, 3 tbl, 9 ex

FIELD: medicine.

SUBSTANCE: invention concerns new biologically active compound peptide possessing protective action against Alzheimer's disease of formula Glu-Trp-Asp-Leu-Val-Gly-Ile-Pro-Gly-Lys-Arg-Ser-Glu-Arg-Phe-Tyr-Glu-Cys-Cys-Lys-Glu.

EFFECT: can be used to produce a preparation for immunoprophylaxis of Alzheimer's diseases.

1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: given invention refers to medicine and concerns a combination for treating an oncological disease, containing as an active ingredient, at least one immunostimulating drug of general formula and one or more chemotherapeutic agents chosen from the group consisting of alkylating agents including carboplatin, cisplatin and oxaliplatin, cytostatic agents chosen from cyclophosphamide and its derivatives, and antimetabolites chosen from fluorouracila and its derivatives, and to a method of treating oncological conditions.

EFFECT: invention provides higher clinical effectiveness.

18 cl, 8 dwg, 5 tbl, 5 ex

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