JP2016531114A - Nutritional composition that inhibits tumor growth - Google Patents

Abstract

The present invention relates to a tumor patient comprising a hydrolyzate of milk protein and fermented milk protein as protein, an oil containing oleic acid as lipid, and milk phospholipid and / or soybean lecithin, and isomaltulose as carbohydrate. The nutrition composition suitable for is provided.

Description

  The present invention relates to nutritional compositions useful for the nutritional management and treatment of tumor patients. The present invention particularly relates to beneficial nutritional compositions for the treatment of patients with colonic solid tumors.

In recent years, it has been reported to treat tumor patients with nutritional compositions. For example, Chinese Patent Application No. CN20061054681 discloses soy milk, vegetable juice, rice flour, wheat and the like.

It is known that soy isoflavone has growth inhibitory effects such as leukemia, colon cancer, lung cancer and stomach cancer.

Chinese Patent Publication No. CN20081006602 discloses a special nutritional food formula containing short peptides and bacteria.

Food prepared according to the food formula enhances immunity, promotes intestinal function, and prevents constipation.

WO2004002238 International publications are nutritional compositions that can reduce the risk of cancer, and these nutritional compositions can induce prebiotic sources and enzyme activity in mammals Combining the additional effects of phytochemicals to reduce the incidence of cancer is disclosed.

In WO2007107295, an internationally publicized publication discloses an invention in which a low GI diet containing a carbohydrate source with a low blood glucose index, such as isomaltulose, lacrosse, trehalulose, and turanose, is effective against malignant tumor diseases for humans and animals. Yes.

In Japanese Patent Application Laid-Open No. 10-203994, a food and beverage composition comprising, as an active ingredient, a peptide having a molecular weight of 500 to 5,000 obtained by allowing a protease to act on one or more of milk, soybean and yeast-derived proteins And inventions applied to cosmetic compositions have been disclosed.

Although these patents and patent applications relate to nutritional compositions useful for the nutritional management and treatment of tumor patients, there are no liquid diet type nutritional compositions of the art type. That is, there are no compositions comprising proteins, lipids, and soluble fibers that supply a certain amount of energy and inulin or a hydrolyzate thereof.

CN20061054681 CN20081006602 WO2004002238 A1 WO2007107295 A1 JP10-203994 JP2001-181303 WO01049308

One object of the present invention is to provide a nutritional composition that inhibits the growth of tumors, particularly colon solid tumors.

The present invention relates to a nutritional composition that reduces tumor weight and tumor volume and inhibits tumor growth so as to obtain smooth edges and high cell growth density in histopathology examinations.

The present invention provides nutritional compositions useful for the nutritional management and treatment of tumor patients. For example, it can be used for nutritional management of cancer patients as liquid food, enteral nutrition, oral / tube feeding, beverage, gel food, and the like. Especially suitable for safe nutrition management of patients with long-term treatment.

The nutritional composition of the present invention can suppress cancer growth at the same time as replenishing nutrition. For example, it can be used for liquid food, oral / enteral / tube feeding, etc. to suppress the growth of cancer in those who have taken it. Moreover, since the composition of this invention can be ingested safely, it is suitable for the nutrition management in long-term medical treatment.

FIG. 1 is a graph showing a survival curve of a mouse having an S-180 type tumor.

FIG. 2 is a graph showing the food intake of a mouse having a C26 type tumor.

FIG. 3 is a graph showing the body weight of a mouse having a C26 type tumor.

FIG. 4 is a photograph of a dissected mouse tumor with a C26 type tumor.

FIG. 5 is a graph showing the tumor volume of a mouse having a C26 type tumor. The

FIG. 6 is a pathological photograph of a tissue of a mouse having a C26 type tumor (Hematoxylin-Eosin Stain, × 100).
BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail. However, the present invention is not limited to the following preferred embodiments, and can be freely changed within the scope of the present invention.

1． protein
1-1 Milk protein hydrolyzate As raw milk protein, casein, whey protein (whey protein concentrate (WPC), whey protein isolate (WPI), α-lactalbumin (α-La), β-lactoglobulin (Β-Lg)), milk protein concentrate (MPC, also called total milk protein (TMP)) and the like can be used.

Taking whey protein hydrolysis as an example, the enzymes used for whey protein hydrolysis are usually pepsin, trypsin and chymotrypsin, but research reports using plant-derived papain, bacteria and fungi-derived proteases (Food Technol 48: 68-71, 1994; Trends Food Sci. Technol., 7: 120-125, 1996; Food Proteins and Their Applications, pp. 443-472, 1997). The activity of an enzyme that hydrolyzes whey protein varies greatly depending on its type. Pepsin degrades α-La and modified α-La, but does not degrade native β-Lg (Neth. Milk dairy J., 47: 15-22, 1993). Trypsin slowly degrades α-La but leaves β-Lg almost undegraded (Neth. Milk dairy J., 45: 225-240, 1991).

Chymotrypsin degrades α-La quickly, but slowly degrades β-Lg.

Papain degrades bovine serum albumin (BSA) and β-Lg, but is difficult to degrade α-La and is resistant to α-La (Int. Dairy Journal 6: 13-31, 1996a). However, papain completely degrades α-La not bound to Ca at an acidic pH (J. Dairy Sci., 76: 311-320, 1993).

  By controlling the degree of milk protein hydrolysis and modifying the milk protein, the functional properties of the milk protein can be altered over a wide range of pH and processing conditions (Enzyme and Chemical Modification of proteins in Food proteins and their Applications, pp. 393-423, 1997, Marcel Dekker, Inc., New York, 1997; Food Technol., 48: 68-71, 1994).

Hydrolysis of peptide bonds results in increased number of charged groups and hydrophobicity, lower molecular weight, and modification of the molecular configuration (J. Dairy Sci., 76: 311-320, 1993). Changes in the functional properties of milk proteins are highly dependent on the degree of hydrolysis.

  For example, the greatest changes commonly seen in whey protein functionality are increased solubility and decreased viscosity. When the degree of hydrolysis of whey protein is high, the hydrolyzate often does not precipitate upon heating and becomes highly soluble at pH 3.5-4.0. Also, the hydrolyzate has a much lower viscosity than intact protein. These differences are particularly noticeable when the protein concentration is high. Other effects include changes in gel properties, improved thermal stability, reduced emulsification and foaming stability (Int. Dairy journal, 6: 13-31, 1996a; Dairy Chemistry 4, pp. 347- 376, 1989; J. Dairy Sci., 79: 782-790, 1996).

Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells (Japanese Patent Laid-Open No. 62-059220).

For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells (Japanese Patent Laid-Open No. 62-059220).

Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells. It is also known that lactoferrin hydrolyzate enhances the cytotoxic activity of antibody drugs in cancer antibody therapy.
Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells. It is also known that lactoferrin hydrolyzate enhances the cytotoxic activity of antibody drugs in cancer antibody therapy.

On the other hand, in the hydrolyzate of milk protein, in addition to the academic literatures exemplified above, there are patent documents (open patent publications and patent publications) regarding many preparation methods. For example, after hydrolyzing casein and whey protein separately, after adsorbing and removing the hydrophobic portion, both are mixed in a predetermined ratio (Japanese Patent No. 2,986,764), and whey protein is combined with a protease derived from the genus Bacillus. After hydrolysis with actinomycete-derived protease, enzyme and insoluble hydrolyzate are removed (Japanese Patent No. 3,222,638), β-lactoglobulin is degraded with enzyme, and branched chain amino acid / aromatic amino acid mole A method for obtaining a mixture of peptides having a ratio of 10% by weight or more, aromatic amino acids of less than 2.0% by weight, and an average molecular weight of several hundred to several thousand (Japanese Patent No. 3,183,945), selective β-lactoglobulin in whey protein Enzymatic degradation method (Japanese Patent No. 2,794,305) or whey protein from B. licheniformis And / or protease from B. subtilis using a non-pH-stat method to hydrolyze to a degree of hydrolysis (DE) of 15-30% and an ultrafiltration exceeding a cut-off value of 10,000 Examples include a method for obtaining a permeate from a membrane (Japanese Patent No. 3167723). The milk protein hydrolyzate of the present invention includes those prepared by methods and techniques other than these patent documents. And

In the hydrolyzate of whey protein, the preparation method which consists of the process of following (1)-(5) can be mentioned.

(1) A whey protein isolate (WPI, Davisco) with a protein content of approximately 90% (w / w) as a dry product is added to distilled water so that the protein concentration is 8% (w / v). Upon dissolution, an aqueous protein solution was obtained.

(2) The aqueous solution was heat treated at 85 ° C. for 2 minutes to denature the protein. The pH of the aqueous solution after this heat treatment was about 7.5.

(3) Thereafter, Alcalase 2.4L (enzyme, Novozymes) was added at 2.0% (w / w) with respect to the concentration of the protein (substrate), and the aqueous solution was maintained at 55 ° C. for 3 hours for hydrolysis. Disassembled.

(4) Next, PTN 6.0S (enzyme, Novozymes Japan), a trypsin derived from swine, was added at a concentration of 3.0% (w / w) with respect to the concentration of the protein (substrate), and the aqueous solution was added. Hydrolysis was carried out by keeping at 3 ° C. for 3 hours. That is, the total hydrolysis time was 6 hours. The pH of the aqueous solution at the end of these hydrolysis reactions was about 7.0.

(5) The hydrolyzate of whey protein was centrifuged (20,000 × g, 10 minutes) and then treated with an ultrafiltration (UF) membrane (Millipore Ultrafree-MC) having a molecular weight cut-off of 10,000.

In the method for preparing a protein hydrolyzate, for example, residual heat, enzyme substrate (E / S), pH, hydrolysis temperature / time, and the like should be considered. Specifically, the residual heat is 65-90 ° C, E / S: 0.01-0.2, hydrolysis temperature: 30-65 ° C, hydrolysis time: 3 hours or more and 20 hours or less.

Examples of the enzyme used for the preparation of the protein hydrolyzate include the followings from Novo Nordisk.
1) End-type protease derived from Bacillus licheniformus: Alcalase
B. lentus origin: Esperase Bacillus subtilis origin: Neutrase
Bacterial origin: Protamex Porcine pancreas origin: PTN (trypsin)
2) Exoprotease Aspergillus oryzae origin: Flavorzyme Pig or bovine viscera origin: Carboxypeptidase

In addition to the above-mentioned enzymes, animal-derived pancreatin, pepsin, plant-derived papain, bromelain, microorganism-derived (for example, lactic acid bacteria, yeast, mold, actinomycetes, etc.) endoprotease and exoprotease, crudely purified products, bacteria Examples include crushed body and the like. Also, as a mixture of enzymes, a combination of Alcalase derived from Bacillus licheniformus and PTN (trypsin) derived from porcine pancreas is often used.

In the protein hydrolyzate of the present invention, the enzyme hydrolyzate itself, a retentate or permeate treated with an ultrafiltration membrane, and the same activity required in the present invention are obtained. It includes certain commercially available milk protein hydrolysates. For example, as the protein hydrolyzate of the present invention, an ultrafiltration membrane retentate having a fractional molecular weight of 10,000 can be used. For example, as the protein hydrolyzate of the present invention, an ultrafiltration membrane retentate having a fractional molecular weight of 10,000 can be used.

The amount of milk protein hydrolyzate in the nutritional composition is the amount of other ingredients (fermented milk protein, fats and oils containing oleic acid, milk phospholipids, soy lecithin, isomaltulose, etc.) It can be adjusted as appropriate depending on the disease state, symptoms, age, weight, use, and the like. Specifically, the amount of milk protein hydrolyzate is 0.9 to 5.0 g, preferably 0.9 to 3.0 g, more preferably 1.0 to 2.5 g, and still more preferably 1.2 to 2.0 g per 100 mL of the nutritional composition. Although it can illustrate, it is not limited to these ranges.

1-2 Fermented milk protein As the origin of the fermented milk protein of the present invention, fermented milk (domestic milk such as cow's milk, buffalo milk, goat milk, sheep milk, horse milk, and / or partially skimmed milk, skimmed milk, reduced product) All liquid milk prepared by combining one or more milk ingredients such as whole milk, reduced skim milk, reduced partially skimmed milk, butter, cream, etc. using a starter such as lactic acid bacteria is used. be able to. For example, fresh cheese, natural cheese, yogurt, and whey cheese are included in the fermented milk of the present invention. The cheese of the present invention is a product obtained by fermenting milk, buttermilk or cream with lactic acid bacteria, or removing whey (whey) from milk, buttermilk or cream, which is obtained by adding an enzyme to the cream, It does not matter whether solidification or aging occurs. Lactobacillus bulgaricus, Streptococcus thermophilus can be mainly used as a starter for producing fermented milk. Lactobacillus acidophilus, Lactobacillus rhamnosis, Lactobacillus plantarum, Lactobacillus murinus, Lactobacillus reuteri, Lactobacillus brevis, Lactobacillus gasseri, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, etc. In addition, microorganisms used in producing fermented milk such as Propionibacterium can be used in combination. The nutritional composition of the present invention may be prepared using any fermented milk, but is preferably prepared using fresh cheese or yoghurt, more preferably quark or yoghurt.

There are many types of fresh cheese such as cottages, quarks, strings, Neuchâtel, cream cheese, mozzarella, ricotta and mascarpone. Quark is a kind of non-ripe-molded (fresh) cheese, characterized by low fat content, refreshing flavor and sourness. A general method for producing quark will be described below. First, the skim milk is sterilized and then inoculated with a lactic acid bacteria starter (mainly Lactobacillus bulgaricus, Streptococcus thermophilus) at 0.5 to 5% (w / w) and fermented. When the pH of the solution reaches 4.6, a curd is formed, and the whey is centrifuged using a quark separator, and then the obtained curd is cooled. As an example of the composition of the quark of the present invention, for example, the total solid content is 17 to 19% (w / w), the protein is 11 to 13% (w / w), the fat is less than 1% (w / w), the carbohydrate 2-8% (w / w) and lactose less than 2% (w / w). In addition, what was solidified using the rennet is also included in the quark of the present invention. Also included in the quark of the present invention are those obtained by adding and cultivating a mixed culture of Lactococcus belonging to Lactococcus, Cremoris and Leuconostoc species to skim milk and removing whey. I decided to. Moreover, what cut | disconnected the whey after heating the solution obtained by cut | disconnecting the card | curd obtained similarly to the said method with a cutter shall also be included in the fresh cheese of this invention.

It is known that lactic acid bacteria and their culture supernatants have an immunoregulatory function. For example, Lactococcus lactis subsp. Cremoris and polysaccharides derived from the culture supernatant thereof are known to be selectively toxic to transformed cells (tumors, cancer cells, etc.) (JP 2009-256312 A). Issue gazette).

The blended amount of fermented milk protein is the blended amount of other ingredients (milk protein hydrolyzate, fat-containing fats and oils containing oleic acid, milk phospholipid, soy lecithin, isomaltulose, etc.) It can be adjusted as appropriate according to symptoms, age, weight, application and the like. Specifically, as a blending amount of fermented milk protein, 0.5 to 6 g, preferably 2 to 6 g, more preferably 2.5 to 4.5 g per 100 mL of nutritional composition in terms of protein can be exemplified. It is not limited to these ranges.

According to the Ministerial Ordinance on Component Standards for Milk and Dairy Products in Japan (Ministry of Health and Welfare Ordinance No. 52 of December 27, 1951), “Standard cattle within five days after distribution” Do not milk milk from goats or sheep. " In other words, this ministerial ordinance restricts the use of colostrum in dairy products.

  In the present invention, it is more preferable to use normal milk (also referred to as mature milk) as the milk used for preparing a milk protein hydrolyzate or fermented milk protein. The origin of milk may be any animal such as cow, buffalo, goat, sheep, horse, human.

2. Lipid
2-1 Use a combination of milk phospholipid and soybean-derived lecithin or egg yolk lecithin as the phospholipid phospholipid. Milk phospholipid alone may be used. The term lecithin is only used for phosphatidylcholine in fields such as biochemistry, medicine and pharmacy, but commercially or industrially it is a mixture of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid and other phospholipids. It is used as a general term. The 7th edition of the Food Additives (1999) defines that lecithin is "obtained from oil seeds or animal raw materials, the main component of which is phospholipid".

Milk phospholipid consists of sphingomyelin (SM), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), lysophosphatidylcholine (LPC), milk fat globule membrane (MFGM) Only localized. The phospholipid fraction of milk fat globule membrane (MFGM) is shown in Table 1 (Bulletin of Japan Dairy Technical Association, Vol. 50, pp. 58-91, 2000).

As shown in Table 1, mickle lecithin contains a particularly large amount of SM but does not contain soy milk lecithin.

Soy lecithin Soy lecithin is a natural food additive and is widely used in the food field, while polyenephosphatidylcholine is also used as a pharmaceutical (indication: improvement of liver function in chronic liver disease, fatty liver, hyperlipidemia) .

The so-called “natural” series of phospholipid products are usually ordered by the PC content in the product. Various types of phospholipids that have been improved according to the use of phospholipids have been produced. For convenience, soy lecithin products are classified as shown in Table 2 due to differences in the main PC content due to the purification and fractionation of soy lecithin (Yamama Fujikawa, Oil Chemistry Vol. 40 (10), pp.951- p58, 1991).

In the present invention, milk phospholipid and soybean lecithin may be used alone or in combination. The amount of milk phospholipid and / or soy lecithin is the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, fats and oils containing oleic acid, isomaltulose, etc.) It can be adjusted as appropriate according to symptoms, age, weight, application and the like. Specifically, the total amount of 0.01-0.5 g, 0.05-0.5 g, 0.1-0.5 g, 0.2-0.3 g per 100 mL of the nutritional composition can be exemplified as the blending amount of milk phospholipid and / or soybean lecithin. However, it is not limited to these ranges.

2-2 Other lipids In the present invention, fats and oils containing oleic acid can be used as lipids. The Ministry of Health, Labor and Welfare has a desirable intake ratio of saturated fatty acids (SFA: palmitic acid, stearic acid, etc.): monounsaturated fatty acids (MUFA: oleic acid, etc.): polyunsaturated fatty acids (PUFA: linoleic acid, linolenic acid, etc.) It is recommended that the ratio of conventional 1: 1.5: 1 to 3: 4: 3 and n-6 fatty acid: n-3 fatty acid ratio be 4: 1. One of the reasons for the recommendation is that in Japan, it is difficult to carry out a diet that increases the intake ratio of MUFA to 1.5 times. Therefore, in order to realize a desirable fatty acid intake ratio (SMP ratio), it is conceivable to increase the content of monounsaturated fatty acids (MUFA) in the fatty acid composition of lipids. Therefore, oleic acid which is a monounsaturated fatty acid can be included in the nutritional composition of the present invention. Examples of lipid sources rich in oleic acid include high oleic sunflower oil, rapeseed oil, olive oil, high oleic safflower oil, soybean oil, corn oil and palm oil. Moreover, nutritionally prepared fats and oils (Nippon Yushi Co., Ltd.) are mentioned as a lipid source containing oleic acid. Sunflower oil, rapeseed oil, olive oil, and mixtures with olive oil can also be used.

The amount of oleic acid is based on the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, milk phospholipid, soy lecithin, isomaltulose, etc.), the condition, symptoms, age, weight, It can be appropriately adjusted depending on the application. Specifically, the blending amount of oleic acid can be exemplified by 25% or more, preferably 30% or more, more preferably 30-50% in the fatty acid composition of the nutritional composition of the present invention. It is not limited to. In addition, polyunsaturated fatty acids such as DHA, EPA and arachidonic acid, medium chain fatty acids such as caprylic acid, capric acid and lauric acid are added, and the ratio of saturated fatty acid: monounsaturated fatty acid: polyunsaturated fatty acid is increased. It can be adjusted to be close to 3: 4: 3.

The amount of oleic acid is based on the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, milk phospholipid, soy lecithin, isomaltulose, etc.), the condition, symptoms, age, weight, It can be appropriately adjusted depending on the application. Specifically, the blending amount of oleic acid can be exemplified by 25% or more, preferably 30% or more, more preferably 30-50% in the fatty acid composition of the nutritional composition of the present invention. It is not limited to. In addition, polyunsaturated fatty acids such as DHA, EPA and arachidonic acid, medium chain fatty acids such as caprylic acid, capric acid and lauric acid are added, and the ratio of saturated fatty acid: monounsaturated fatty acid: polyunsaturated fatty acid is increased. It can be adjusted to be close to 3: 4: 3.

3.3 As a carbohydrate and dietary fiber, isomaltulose can be mainly used. Examples of other saccharides include sugar alcohols (such as sorbitol, xylitol, maltitol), honey, granulated sugar, glucose, fructose, and invert sugar.
Isomaltulose is a disaccharide in which glucose and fructose molecules are linked together by α-1,6 molecules, and is a structural isomer of sucrose, which is 6-O- (α-D-Glucopyranosyl) -D-fructose or isoform. Also called maltulose or palatinose. It has a molecular weight of 342.297 and Cas. No. 13718-94-0 and is used as a sweetener. Isomaltulose is contained in very small amounts in honey and sugarcane.

In addition, α-glucosyltransferase derived from Protaminobacter rubrum is allowed to act on sucrose to transfer α-1,2 bonds to α-1,6 bonds to produce isomaltulose. You can also The sweetness of isomaltulose is similar to that of sucrose, but the sweetness is about half that of sucrose. Orally ingested isomaltulose is degraded by isomaltase in the digestive tract, and is digested and absorbed into glucose and fructose in the same way as sucrose (Toshinao Aida, Journal of Japanese Society of Nutrition and Food, Vol. 36 (3): 169-173, 1983). In addition, isomaltose, panose, isomalttriose and the like that are digested with isomaltase compete with the digestion of isomaltulose, and it is said that digestive absorption is suppressed by ingesting isomaltulose ( Japanese Journal of Nutrition and Food, 36 (3), pp.169-173 (1983)). The calorie of isomaltulose is 4kcal / g. In the present invention, isomaltulose includes palatinose syrup, reduced palatinose or palatinose syrup. Palatinose starch syrup is a starch syrup-like liquid substance mainly composed of oligosaccharides such as tetrasaccharide, hexasaccharide and octasaccharide produced by dehydration condensation of isomaltulose.

With respect to isomaltulose, it is known that a low glycemic food composition containing isomaltulose or the like is used for the treatment / prevention of malignant tumor diseases (Japanese Patent Publication No. 2009-530326).

The blending amount of isomaltulose is blended with other ingredients (milk protein hydrolyzate, fermented milk protein, fats and oils containing oleic acid, milk phospholipids, soybean lecithin, etc.) It can be adjusted as appropriate according to age, weight, use and the like. Specifically, the blending amount of isomaltulose can be exemplified by 4 to 15 g, preferably 5 to 7 g per 100 mL of the nutritional composition, but is not limited to these ranges (Special Table No. 2009-530326) Publication).

The amount of heat of the nutritional composition of the present invention can be adjusted by appropriately adding proteins, lipids and carbohydrates. The calorie | heat amount of the nutrition composition of this invention can illustrate 50-150 kcal per 100 ml of nutrition composition, Preferably, it is 80-120 kcal, However, It is not limited to these ranges.

In addition, the energy ratio of the protein, lipid, and carbohydrate to the whole nutritional composition in the nutritional composition of the present invention substantially conforms to the nutritional requirements of the sixth revised Japanese. Specific examples include 15 to 25% protein, 20 to 30% lipid, and 45 to 65% carbohydrate. However, the present invention is not limited to these ranges.

The nutritional composition of the present invention may also contain dietary fiber. Dietary fiber refers to substances in food that are not hydrolyzed by human digestive enzymes, and is classified into water-soluble dietary fiber and insoluble dietary fiber based on their affinity for water. As water-soluble dietary fiber, indigestible oligosaccharides such as lactulose, lactitol, or raffinose can be used. Physiological functions of indigestible oligosaccharides are known to reach the large intestine as undigested substances, contribute to the activation and proliferation of intestinal bifidobacteria, and have an improved intestinal environment, ie, an intestinal regulating effect. Yes.

Furthermore, as water-soluble dietary fiber candidates, high-molecular water-soluble dietary fiber includes soybean thickening polysaccharide, konjac glucomannan, alginic acid, low molecular alginic acid, psyllium, gum arabic, seaweed polysaccharide (cellulose, lignin-like substance, agar). , Carrageenan, alginic acid, fucodyne, laminarin), microbial gum (welan gum, curdlan, xanthan gum, gellan gum, dextran, pullulan, lambzan gum), other gums (seed-derived locust bean gum, tamarind gum, tara gum, sap-derived caraya gum, Polydextrose of low molecular weight water-soluble dietary fiber such as tragacanth gum), indigestible dextrin, pine fiber, maltitol and the like can be used.

Insoluble dietary fiber increases the mass of indigestibles in the large intestine and shortens transit time. As a result, the number of defecations increases, resulting in an increase in stool volume. Insoluble dietary fiber candidates include cellulose, hemicellulose, lignin, chitin, chitosan, soy bran, wheat bran, barley bran, pine fiber, corn fiber, beet fiber, oat bran, rye bran, pearl bran, rice bran, millet, millet Millet bran such as Japanese millet, sorghum, cereal bran, legume bran, sesame bran, and okara. Oat bran, rye bran, barley bran, rice bran, millet, millet, millet, sorghum bran, cereal bran, buckwheat bran, sesame bran, and okara.

The nutritional composition of the present invention includes water, protein, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors, emulsifiers in addition to the proteins, lipids, carbohydrates, and dietary fibers. , Thickeners, stabilizers and the like can be used. Examples of the protein include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, whey protein hydrolysate, α-casein, β-casein, κ- Casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and vegetable proteins, their degradation products; butter, whey minerals, cream, whey, non-protein nitrogen And various milk-derived components such as sialic acid, phospholipid, and lactose. It may contain peptides such as casein phosphopeptides and lysines and amino acids. Examples of the saccharide include saccharides, processed starch (in addition to text phosphorus, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of lipids include animal oils such as lard and fish oil, fractionated oils thereof, hydrogenated oils and transesterified oils; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, tartaric acid, erythorbic acid, and the like. In addition, a material containing a fecal odor reducing effect (for example, champignon extract 5 mg to 500 mg (0.005% to 0.5%)), a carotenoid preparation (for example, α-carotene, β-carotene, lycopene, lutein, etc.) 10 μg to 200 μg (0.00001% to 0.0002%)) and an antioxidant (catechin, polyphenol, etc.) can also be included. Two or more of these components can be used in combination, and a synthetic product and / or a food containing a large amount thereof may be used. The form of the food may be any form such as a solid, liquid, or gel.

  The nutritional composition of the present invention can be produced by methods known in the art. After some or all of the raw materials have been prepared, homogenization is performed as necessary. Homogenization means homogenization by thoroughly mixing each prepared component, and the fat globules and coarse particles of other components are mechanically refined to prevent the rising and aggregation of fat and the like, and the nutritional composition It means making into a uniform emulsified state.

In the production of the nutritional composition of the present invention, heat treatment or heat sterilization is performed. As heat sterilization conditions, general food sterilization conditions can be used, and heat sterilization can be performed using a conventional apparatus. For example, sterilization of 62-65 ° C. × 30 minutes, 72 ° C. or more × 15 seconds or more, 72 ° C. or more × 15 minutes or more, or 120-150 ° C. × 1-5 seconds, or 121-124 ° C. × 5-20 minutes, 105 Although sterilization of ˜140 ° C., retort (pressure heating) sterilization, high-pressure steam sterilization, and the like can be used, it is not limited to these examples. The heat sterilization can be preferably performed under pressure.

In addition, after preliminarily heat-sterilizing the liquid nutritional composition, aseptically filling the container (for example, a method using a combination of the UHT sterilization method and the aseptic packaging method), filling the container with the liquid nutritional composition, Method of heat sterilization with containers (for example, autoclave method), can containers, liquid foods, various containers used for oral and tube feeding (so-called soft bags, nutrition bags, etc.) and retort sterilization (for example, at 115 to 145 ° C 5 ~ 10 seconds), filling in cans, liquid foods and various containers used for oral and tube feeding (so-called soft bags, nutrition bags, etc.) and sterilized by retort for about 5-8 seconds at about 140-145 ° C Although the method of cooling after heat sterilization and performing aseptic filling can be illustrated, it is not limited to these examples.

By the heat treatment or heat sterilization, starters (such as lactic acid bacteria, bifidobacteria, or Propionibacterium spp.) Derived from fermented milk as a raw material are killed.

The composition of the present invention can be used to suppress the growth of cancer. The nutritional composition of the present invention can be used for nutritional management of oral and enteral nutrition patients, the elderly, infants and the like, for example, as liquid foods, oral and tube feedings, beverages, gel foods and the like.

The osmotic pressure of the nutritional composition of the present invention can be exemplified by an osmotic pressure of about 500 to 1000 mOsm / l, such as about 550 to 750 mOsm / l. When measured at room temperature, the viscosity of the nutritional composition can be exemplified by 20-100 cp (1 cp = 1 mPa · s), preferably 25-60 cp, more preferably 30-50 cp. The range is not limited.
The pH of the nutritional composition of the present invention can be adjusted to 4.6 or less, preferably 3.0 to 4.3, more preferably 3.8 to 4.2, but is not limited to these ranges.

The nutritional composition of the present invention is beneficial in inhibiting tumor growth in tumor patients. In particular, colon tumor patients, colon solid tumor patients, sarcoma cell tumor patients, lung cancer patients, thymoma patients, breast cancer patients, pharyngeal cancer patients, laryngeal cancer patients, gastric cancer patients, esophageal cancer patients, hepatocellular carcinoma patients, cholangiocarcinoma patients, Gallbladder cancer patients, pancreatic cancer patients, penile cancer patients, renal pelvis and ureter cancer patients, renal cell cancer patients, testicular cancer patients, prostate cancer patients, bladder cancer patients, vulvar cancer patients, cervical cancer patients, endometrial cancer patients It is useful for inhibiting the growth of tumors such as uterine sarcoma patients, vaginal cancer patients, ovarian cancer patients, ovarian germ cell tumor patients, or brain tumor patients.

In an in vivo test using a mouse transplanted with a tumor cell line, it was confirmed that the nutritional composition of the present invention inhibits tumor growth and tumor infiltration into surrounding tissues. When tumors invade surrounding tissues, tumor metastasis is likely to occur. In addition, pain occurs when a tumor infiltrates an internal organ or peripheral nerve. Therefore, the composition of the present invention can suppress tumor growth, tumor invasion / metastasis, or tumor-induced pain.

The daily intake of the nutritional composition of the present invention in a pharmaceutical product or food / drink depends on the pathology, age, symptoms, weight, use of the subject, and whether the nutritional composition of the present invention is the only nutritional item. Since it differs, it is not specifically limited. Specifically, in the case of an adult, the intake per day may be 400 to 1600 ml, preferably 600 to 1600 ml, more preferably 800 to 1200 ml. For adults, for example, up to 3000 ml per day may be taken. The intake amount can also be determined by the attending physician of the subject person.
Moreover, you may use the nutrition composition of this invention together with the pharmaceutical and foodstuff which have an antitumor action conventionally known.

The nutritional composition of the present invention can be used in the form of any medicine or food or drink. For example, the intestinal flora can be obtained by directly administering the nutritional composition of the present invention as a pharmaceutical or by directly ingesting it as a special-purpose food such as a food for specified health use, a nutritional functional food, a nutritional supplement, a liquid food or a supplement. Is expected to improve. In addition, various foods (milk, soft drinks, fermented milk, yogurt, cheese, bread, biscuits, crackers, pizza crusts, prepared milk powder, liquid foods, special-purpose foods, regardless of the form of liquid, paste, solid, powder, etc. , Foods for the sick, nutritional foods, frozen foods, processed foods, and other commercially available foods). Moreover, when the usage form of a nutrient composition is a powder, it can manufacture by using means, such as spray drying and freeze-drying, for example.

When the nutritional composition of the present invention is used as a pharmaceutical or a supplement, it can be administered in various forms. Examples of the form include oral administration using tablets, capsules, granules, powders, syrups and the like. These various preparations can be generally used in the pharmaceutical preparation technical field such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizers, suspension agents, coating agents, etc. as main ingredients according to conventional methods. It can be formulated using known adjuvants for formulation. Further, it may contain an appropriate amount of calcium. Further, an appropriate amount of vitamins, minerals, organic acids, sugars, amino acids, peptides, etc. may be added.
Example

Hereinafter, although an example is given and explained about the present invention, the present invention is not limited by this.
Materials and methods

Two commonly used cancer cell models, mouse sarcoma cells (S180) and mouse colon solid tumor cells (C26), construct S-180 as an ascites tumor model and C-26 as a solid tumor model. Randomly divide the mice into two groups so that half are male and half are female. The group fed with the nutritional composition of the present invention with 14 days of constant feeding is called “test composition of the present invention” and AIN-93M fed with the control diet. The survival time of mice bearing S-180 tumors was recorded as the main index, and food intake, body weight, tumor weight, and tumor volume of mice bearing C-26 tumors were recorded. Serum TNF-a and IL-6 levels were measured and tumor histopathology was also performed.
1. Meal test

  The test composition (yellow fine powder) of the present invention is a dried liquid food composition shown in Tables 1 and 3. The total fatty acid composition is formed with 39% by weight of liquid oleic acid, 1.2% by weight of eicosapentaenoic acid and 0.8% by weight of docosahexaenoic acid, and AIN-93M (white fine powder) is manufactured by Oriental Yeast Industry (Japan). A diet refined to the AIN-93M standard, commonly used in nutrition experiments in rats and mice, and its ingredients were published in 1993 by the United States National Institute of Nutrition (Table 4). And Table 5).

2. Transplanted tumor cell model
S-180 cells were provided by the Shanghai Institute of Oncology.
C-26 cells were provided by the Shanghai Institute of Oncology.
3. animal

Variety: BALB / c mice
Source: Shanghai Shanghai SIPPR-BK Laboratory Animal Co. Ltd
Gender: Half male and half female Age: 6-7 weeks Weight: 18-20g
Number: 76 (36 mice for S-180 experiments, 40 mice for C26 experiments)
Animal serial number: SCXK (Shanghai) 2008-0016
Animal license number: SYXK (Shanghai) 2008-0027
Storage conditions: SPF Animal Experiment House, room temperature 24 + 1 ° C, humidity 40-70%, 12/12 hours and night cycle.

As a commonly used mouse cancer model, ascites tumor model S-180 and solid tumor model C-26 are used to determine whether the test composition of the present invention can inhibit tumor growth. evaluate. As a result, in the test set of the present invention, the mean survival time of the mice having S-180 tumor was not significantly different from the control mice (P> O. 05). However, the test set of the present invention was able to inhibit tumor growth in the C-26 mouse tumor model.

INDUSTRIAL APPLICABILITY The nutritional composition of the present invention is beneficial to colon solid tumor patients by inhibiting the growth of colon solid tumors or reducing the invasion of malignant tumors.

  The present invention relates to nutritional compositions useful for the nutritional management and treatment of tumor patients. The present invention particularly relates to beneficial nutritional compositions for the treatment of patients with colonic solid tumors.

In recent years, it has been reported to treat tumor patients with nutritional compositions. For example, Chinese Patent Application No. CN20061054681 discloses soy milk, vegetable juice, rice flour, wheat and the like.

It is known that soy isoflavone has growth inhibitory effects such as leukemia, colon cancer, lung cancer and stomach cancer.

Chinese Patent Publication No. CN20081006602 discloses a special nutritional food formula containing short peptides and bacteria.

Food prepared according to the food formula enhances immunity, promotes intestinal function, and prevents constipation.

WO2004002238 International publications are nutritional compositions that can reduce the risk of cancer, and these nutritional compositions can induce prebiotic sources and enzyme activity in mammals Combining the additional effects of phytochemicals to reduce the incidence of cancer is disclosed.

In WO2007107295, an internationally publicized publication discloses an invention in which a low GI diet containing a carbohydrate source with a low blood glucose index, such as isomaltulose, lacrosse, trehalulose, and turanose, is effective against malignant tumor diseases for humans and animals. Yes.

In Japanese Patent Application Laid-Open No. 10-203994, a food and beverage composition comprising, as an active ingredient, a peptide having a molecular weight of 500 to 5,000 obtained by allowing a protease to act on one or more of milk, soybean and yeast-derived proteins And inventions applied to cosmetic compositions have been disclosed.

Although these patents and patent applications relate to nutritional compositions useful for the nutritional management and treatment of tumor patients, there are no liquid diet type nutritional compositions of the art type. That is, there are no compositions comprising proteins, lipids, and soluble fibers that supply a certain amount of energy and inulin or a hydrolyzate thereof.

CN20061054681 CN20081006602 WO2004002238 A1 WO2007107295 A1 JP10-203994 JP2001-181303 WO01049308

One object of the present invention is to provide a nutritional composition that inhibits the growth of tumors, particularly colon solid tumors.

The present invention relates to a nutritional composition that reduces tumor weight and tumor volume and inhibits tumor growth so as to obtain smooth edges and high cell growth density in histopathology examinations.

The present invention provides nutritional compositions useful for the nutritional management and treatment of tumor patients. For example, it can be used for nutritional management of cancer patients as liquid food, enteral nutrition, oral / tube feeding, beverage, gel food, and the like. Especially suitable for safe nutrition management of patients with long-term treatment.

The nutritional composition of the present invention can suppress cancer growth at the same time as replenishing nutrition. For example, it can be used for liquid food, oral / enteral / tube feeding, etc. to suppress the growth of cancer in those who have taken it. Moreover, since the composition of this invention can be ingested safely, it is suitable for nutrition management in long-term medical treatment.

FIG. 1 is a graph showing a survival curve of a mouse having an S-180 type tumor.

FIG. 2 is a graph showing the food intake of a mouse having a C26 type tumor.

FIG. 3 is a graph showing the body weight of a mouse having a C26 type tumor.

FIG. 4 is a photograph of a dissected mouse tumor with a C26 type tumor.

FIG. 5 is a graph showing the tumor volume of a mouse having a C26 type tumor. The

FIG. 6 is a pathological photograph of a tissue of a mouse having a C26 type tumor (Hematoxylin-Eosin Stain, × 100).
BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail. However, the present invention is not limited to the following preferred embodiments, and can be freely changed within the scope of the present invention.

1． protein
1-1 Milk protein hydrolyzate As raw milk protein, casein, whey protein (whey protein concentrate (WPC), whey protein isolate (WPI), α-lactalbumin (α-La), β-lactoglobulin (Β-Lg)), milk protein concentrate (MPC, also called total milk protein (TMP)) and the like can be used.

Taking whey protein hydrolysis as an example, the enzymes used for whey protein hydrolysis are usually pepsin, trypsin and chymotrypsin, but research reports using plant-derived papain, bacteria and fungi-derived proteases (Food Technol 48: 68-71, 1994; Trends Food Sci. Technol., 7: 120-125, 1996; Food Proteins and Their Applications, pp. 443-472, 1997). The activity of an enzyme that hydrolyzes whey protein varies greatly depending on its type. Pepsin degrades α-La and modified α-La, but does not degrade native β-Lg (Neth. Milk dairy J., 47: 15-22, 1993). Trypsin slowly degrades α-La but leaves β-Lg almost undegraded (Neth. Milk dairy J., 45: 225-240, 1991).

Chymotrypsin degrades α-La quickly, but slowly degrades β-Lg.

Papain degrades bovine serum albumin (BSA) and β-Lg, but is difficult to degrade α-La and is resistant to α-La (Int. Dairy Journal 6: 13-31, 1996a). However, papain completely degrades α-La not bound to Ca at an acidic pH (J. Dairy Sci., 76: 311-320, 1993).

  By controlling the degree of milk protein hydrolysis and modifying the milk protein, the functional properties of the milk protein can be altered over a wide range of pH and processing conditions (Enzyme and Chemical Modification of proteins in Food proteins and their Applications, pp. 393-423, 1997, Marcel Dekker, Inc., New York, 1997; Food Technol., 48: 68-71, 1994).

Hydrolysis of peptide bonds results in increased number of charged groups and hydrophobicity, lower molecular weight, and modification of the molecular configuration (J. Dairy Sci., 76: 311-320, 1993). Changes in the functional properties of milk proteins are highly dependent on the degree of hydrolysis.

  For example, the greatest changes commonly seen in whey protein functionality are increased solubility and decreased viscosity. When the degree of hydrolysis of whey protein is high, the hydrolyzate often does not precipitate upon heating and becomes highly soluble at pH 3.5-4.0. Also, the hydrolyzate has a much lower viscosity than intact protein. These differences are particularly noticeable when the protein concentration is high. Other effects include changes in gel properties, improved thermal stability, reduced emulsification and foaming stability (Int. Dairy journal, 6: 13-31, 1996a; Dairy Chemistry 4, pp. 347- 376, 1989; J. Dairy Sci., 79: 782-790, 1996).

Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells (Japanese Patent Laid-Open No. 62-059220).

For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells (Japanese Patent Laid-Open No. 62-059220).

Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells. It is also known that lactoferrin hydrolyzate enhances the cytotoxic activity of antibody drugs in cancer antibody therapy.
Various physiologically active peptides derived from milk proteins are known. For example, it is known that autolysates of protein digestive enzymes such as pepsin suppress cancer metastasis through the platelet aggregation-inducing activity of cancer cells. It is also known that lactoferrin hydrolyzate enhances the cytotoxic activity of antibody drugs in cancer antibody therapy.

On the other hand, in the hydrolyzate of milk protein, in addition to the academic literatures exemplified above, there are patent documents (open patent publications and patent publications) regarding many preparation methods. For example, after hydrolyzing casein and whey protein separately, after adsorbing and removing the hydrophobic portion, both are mixed in a predetermined ratio (Japanese Patent No. 2,986,764), and whey protein is combined with a protease derived from the genus Bacillus. After hydrolysis with actinomycete-derived protease, enzyme and insoluble hydrolyzate are removed (Japanese Patent No. 3,222,638), β-lactoglobulin is degraded with enzyme, and branched chain amino acid / aromatic amino acid mole A method for obtaining a mixture of peptides having a ratio of 10% by weight or more, aromatic amino acids of less than 2.0% by weight, and an average molecular weight of several hundred to several thousand (Japanese Patent No. 3,183,945), selective β-lactoglobulin in whey protein Enzymatic degradation method (Japanese Patent No. 2,794,305) or whey protein from B. licheniformis And / or protease from B. subtilis using a non-pH-stat method to hydrolyze to a degree of hydrolysis (DE) of 15-30% and an ultrafiltration exceeding a cut-off value of 10,000 Examples include a method for obtaining a permeate from a membrane (Japanese Patent No. 3167723). The milk protein hydrolyzate of the present invention includes those prepared by methods and techniques other than these patent documents. And

In the hydrolyzate of whey protein, the preparation method which consists of the process of following (1)-(5) can be mentioned.

(1) A whey protein isolate (WPI, Davisco) with a protein content of approximately 90% (w / w) as a dry product is added to distilled water so that the protein concentration is 8% (w / v). Upon dissolution, an aqueous protein solution was obtained.

(2) The aqueous solution was heat treated at 85 ° C. for 2 minutes to denature the protein. The pH of the aqueous solution after this heat treatment was about 7.5.

(3) Thereafter, Alcalase 2.4L (enzyme, Novozymes) was added at 2.0% (w / w) with respect to the concentration of the protein (substrate), and the aqueous solution was maintained at 55 ° C. for 3 hours for hydrolysis. Disassembled.

(4) Next, PTN 6.0S (enzyme, Novozymes Japan), a trypsin derived from swine, was added at a concentration of 3.0% (w / w) with respect to the concentration of the protein (substrate), and the aqueous solution was 55%. Hydrolysis was carried out by keeping at 3 ° C. for 3 hours. That is, the total hydrolysis time was 6 hours. The pH of the aqueous solution at the end of these hydrolysis reactions was about 7.0.

(5) The hydrolyzate of whey protein was centrifuged (20,000 × g, 10 minutes) and then treated with an ultrafiltration (UF) membrane (Millipore Ultrafree-MC) having a molecular weight cut-off of 10,000.

In the method for preparing a protein hydrolyzate, for example, residual heat, enzyme substrate (E / S), pH, hydrolysis temperature / time, and the like should be considered. Specifically, the residual heat is 65-90 ° C, E / S: 0.01-0.2, hydrolysis temperature: 30-65 ° C, hydrolysis time: 3 hours or more and 20 hours or less.

Examples of the enzyme used for the preparation of the protein hydrolyzate include the followings from Novo Nordisk.
1) End-type protease derived from Bacillus licheniformus: Alcalase
B. lentus origin: Esperase Bacillus subtilis origin: Neutrase
Bacterial origin: Protamex Porcine pancreas origin: PTN (trypsin)
2) Exoprotease Aspergillus oryzae origin: Flavorzyme Pig or bovine viscera origin: Carboxypeptidase

In addition to the above-mentioned enzymes, animal-derived pancreatin, pepsin, plant-derived papain, bromelain, microorganism-derived (for example, lactic acid bacteria, yeast, mold, actinomycetes, etc.) endoprotease and exoprotease, crudely purified products, bacteria Examples include crushed body and the like. Also, as a mixture of enzymes, a combination of Alcalase derived from Bacillus licheniformus and PTN (trypsin) derived from porcine pancreas is often used.

In the protein hydrolyzate of the present invention, the enzyme hydrolyzate itself, a retentate or permeate treated with an ultrafiltration membrane, and the same activity required in the present invention are obtained. It includes certain commercially available milk protein hydrolysates. For example, as the protein hydrolyzate of the present invention, an ultrafiltration membrane retentate having a fractional molecular weight of 10,000 can be used. For example, as the protein hydrolyzate of the present invention, an ultrafiltration membrane retentate having a fractional molecular weight of 10,000 can be used.

The amount of milk protein hydrolyzate in the nutritional composition is the amount of other ingredients (fermented milk protein, fats and oils containing oleic acid, milk phospholipids, soy lecithin, isomaltulose, etc.) It can be adjusted as appropriate depending on the disease state, symptoms, age, weight, use, and the like. Specifically, the amount of milk protein hydrolyzate is 0.9 to 5.0 g, preferably 0.9 to 3.0 g, more preferably 1.0 to 2.5 g, and still more preferably 1.2 to 2.0 g per 100 mL of the nutritional composition. Although it can illustrate, it is not limited to these ranges.

1-2 Fermented milk protein As the origin of the fermented milk protein of the present invention, fermented milk (domestic milk such as cow's milk, buffalo milk, goat milk, sheep milk, horse milk, and / or partially skimmed milk, skimmed milk, reduced product) All liquid milk prepared by combining one or more milk ingredients such as whole milk, reduced skim milk, reduced partially skimmed milk, butter, cream, etc. using a starter such as lactic acid bacteria is used. be able to. For example, fresh cheese, natural cheese, yogurt, and whey cheese are included in the fermented milk of the present invention. The cheese of the present invention is a product obtained by fermenting milk, buttermilk or cream with lactic acid bacteria, or removing whey (whey) from milk, buttermilk or cream, which is obtained by adding an enzyme to the cream, It does not matter whether solidification or aging occurs. Lactobacillus bulgaricus, Streptococcus thermophilus can be mainly used as a starter for producing fermented milk. Lactobacillus acidophilus, Lactobacillus rhamnosis, Lactobacillus plantarum, Lactobacillus murinus, Lactobacillus reuteri, Lactobacillus brevis, Lactobacillus gasseri, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, etc. In addition, microorganisms used in producing fermented milk such as Propionibacterium can be used in combination. The nutritional composition of the present invention may be prepared using any fermented milk, but is preferably prepared using fresh cheese or yoghurt, more preferably quark or yoghurt.

There are many types of fresh cheese such as cottages, quarks, strings, Neuchâtel, cream cheese, mozzarella, ricotta and mascarpone. Quark is a kind of non-ripe-molded (fresh) cheese, characterized by low fat content, refreshing flavor and sourness. A general method for producing quark will be described below. First, the skim milk is sterilized and then inoculated with a lactic acid bacteria starter (mainly Lactobacillus bulgaricus, Streptococcus thermophilus) at 0.5 to 5% (w / w) and fermented. When the pH of the solution reaches 4.6, a curd is formed, and the whey is centrifuged using a quark separator, and then the obtained curd is cooled. As an example of the composition of the quark of the present invention, for example, the total solid content is 17 to 19% (w / w), the protein is 11 to 13% (w / w), the fat is less than 1% (w / w), the carbohydrate 2-8% (w / w) and lactose less than 2% (w / w). In addition, what was solidified using the rennet is also included in the quark of the present invention. Also included in the quark of the present invention are those obtained by adding and cultivating a mixed culture of Lactococcus belonging to Lactococcus, Cremoris and Leuconostoc species to skim milk and removing whey. I decided to. Moreover, what cut | disconnected the whey after heating the solution obtained by cut | disconnecting the card | curd obtained similarly to the said method with a cutter shall also be included in the fresh cheese of this invention.

It is known that lactic acid bacteria and their culture supernatants have an immunoregulatory function. For example, Lactococcus lactis subsp. Cremoris and polysaccharides derived from the culture supernatant thereof are known to be selectively toxic to transformed cells (tumors, cancer cells, etc.) (JP 2009-256312 A). Issue gazette).

The blended amount of fermented milk protein is the blended amount of other ingredients (milk protein hydrolyzate, fat-containing fats and oils containing oleic acid, milk phospholipid, soy lecithin, isomaltulose, etc.) It can be adjusted as appropriate according to symptoms, age, weight, application and the like. Specifically, as a blending amount of fermented milk protein, 0.5 to 6 g, preferably 2 to 6 g, more preferably 2.5 to 4.5 g per 100 mL of nutritional composition in terms of protein can be exemplified. It is not limited to these ranges.

According to the Ministerial Ordinance on Component Standards for Milk and Dairy Products in Japan (Ministry of Health and Welfare Ordinance No. 52 of December 27, 1951), “Standard cattle within five days after distribution” Do not milk milk from goats or sheep. " In other words, this ministerial ordinance restricts the use of colostrum in dairy products.

  In the present invention, it is more preferable to use normal milk (also referred to as mature milk) as the milk used for preparing a milk protein hydrolyzate or fermented milk protein. The origin of milk may be any animal such as cow, buffalo, goat, sheep, horse, human.

2. Lipid
2-1 Use a combination of milk phospholipid and soybean-derived lecithin or egg yolk lecithin as the phospholipid phospholipid. Milk phospholipid alone may be used. The term lecithin is only used for phosphatidylcholine in fields such as biochemistry, medicine and pharmacy, but commercially or industrially it is a mixture of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid and other phospholipids. It is used as a general term. The 7th edition of the Food Additives (1999) defines that lecithin is "obtained from oil seeds or animal raw materials, the main component of which is phospholipid".

Milk phospholipid consists of sphingomyelin (SM), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), lysophosphatidylcholine (LPC), milk fat globule membrane (MFGM) Only localized. The phospholipid fraction of milk fat globule membrane (MFGM) is shown in Table 1 (Bulletin of Japan Dairy Technical Association, Vol. 50, pp. 58-91, 2000).

As shown in Table 1, mickle lecithin contains a particularly large amount of SM but does not contain soy milk lecithin.

Soy lecithin Soy lecithin is a natural food additive and is widely used in the food field, while polyenephosphatidylcholine is also used as a pharmaceutical (indication: improvement of liver function in chronic liver disease, fatty liver, hyperlipidemia) .

The so-called “natural” series of phospholipid products are usually ordered by the PC content in the product. Various types of phospholipids that have been improved according to the use of phospholipids have been produced. For convenience, soy lecithin products are classified as shown in Table 2 due to differences in the main PC content due to the purification and fractionation of soy lecithin (Yamama Fujikawa, Oil Chemistry Vol. 40 (10), pp.951- p58, 1991).

In the present invention, milk phospholipid and soybean lecithin may be used alone or in combination. The amount of milk phospholipid and / or soy lecithin is the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, fats and oils containing oleic acid, isomaltulose, etc.) It can be adjusted as appropriate according to symptoms, age, weight, application and the like. Specifically, the total amount of 0.01-0.5 g, 0.05-0.5 g, 0.1-0.5 g, 0.2-0.3 g per 100 mL of the nutritional composition can be exemplified as the blending amount of milk phospholipid and / or soybean lecithin. However, it is not limited to these ranges.

2-2 Other lipids In the present invention, fats and oils containing oleic acid can be used as lipids. The Ministry of Health, Labor and Welfare has a desirable intake ratio of saturated fatty acids (SFA: palmitic acid, stearic acid, etc.): monounsaturated fatty acids (MUFA: oleic acid, etc.): polyunsaturated fatty acids (PUFA: linoleic acid, linolenic acid, etc.) It is recommended that the ratio of conventional 1: 1.5: 1 to 3: 4: 3 and n-6 fatty acid: n-3 fatty acid ratio be 4: 1. One of the reasons for the recommendation is that in Japan, it is difficult to carry out a diet that increases the intake ratio of MUFA to 1.5 times. Therefore, in order to realize a desirable fatty acid intake ratio (SMP ratio), it is conceivable to increase the content of monounsaturated fatty acids (MUFA) in the fatty acid composition of lipids. Therefore, oleic acid which is a monounsaturated fatty acid can be included in the nutritional composition of the present invention. Examples of lipid sources rich in oleic acid include high oleic sunflower oil, rapeseed oil, olive oil, high oleic safflower oil, soybean oil, corn oil and palm oil. Moreover, nutritionally prepared fats and oils (Nippon Yushi Co., Ltd.) are mentioned as a lipid source containing oleic acid. Sunflower oil, rapeseed oil, olive oil, and mixtures with olive oil can also be used.

The amount of oleic acid is based on the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, milk phospholipid, soy lecithin, isomaltulose, etc.), the condition, symptoms, age, weight, It can be appropriately adjusted depending on the application. Specifically, the blending amount of oleic acid can be exemplified by 25% or more, preferably 30% or more, more preferably 30-50% in the fatty acid composition of the nutritional composition of the present invention. It is not limited to. In addition, polyunsaturated fatty acids such as DHA, EPA and arachidonic acid, medium chain fatty acids such as caprylic acid, capric acid and lauric acid are added, and the ratio of saturated fatty acid: monounsaturated fatty acid: polyunsaturated fatty acid is increased. It can be adjusted to be close to 3: 4: 3.

The amount of oleic acid is based on the amount of other ingredients (milk protein hydrolyzate, fermented milk protein, milk phospholipid, soy lecithin, isomaltulose, etc.), the condition, symptoms, age, weight, It can be appropriately adjusted depending on the application. Specifically, the blending amount of oleic acid can be exemplified by 25% or more, preferably 30% or more, more preferably 30-50% in the fatty acid composition of the nutritional composition of the present invention. It is not limited to. In addition, polyunsaturated fatty acids such as DHA, EPA and arachidonic acid, medium chain fatty acids such as caprylic acid, capric acid and lauric acid are added, and the ratio of saturated fatty acid: monounsaturated fatty acid: polyunsaturated fatty acid is increased. It can be adjusted to be close to 3: 4: 3.

3.3 As a carbohydrate and dietary fiber, isomaltulose can be mainly used. Examples of other saccharides include sugar alcohols (such as sorbitol, xylitol, maltitol), honey, granulated sugar, glucose, fructose, and invert sugar.
Isomaltulose is a disaccharide in which glucose and fructose molecules are linked together by α-1,6, and is a structural isomer of sucrose. Also called maltulose or palatinose. It has a molecular weight of 342.297 and Cas. No. 13718-94-0 and is used as a sweetener. Isomaltulose is contained in very small amounts in honey and sugarcane.

In addition, α-glucosyltransferase derived from Protaminobacter rubrum is allowed to act on sucrose to transfer α-1,2 bonds to α-1,6 bonds to produce isomaltulose. You can also The sweetness of isomaltulose is similar to that of sucrose, but the sweetness is about half that of sucrose. Orally ingested isomaltulose is degraded by isomaltase in the digestive tract, and is digested and absorbed into glucose and fructose in the same way as sucrose (Toshinao Aida, Journal of Japanese Society of Nutrition and Food, Vol. 36 (3): 169-173, 1983). In addition, isomaltose, panose, isomalttriose and the like that are digested with isomaltase compete with the digestion of isomaltulose, and it is said that digestive absorption is suppressed by ingesting isomaltulose ( Japanese Journal of Nutrition and Food, 36 (3), pp.169-173 (1983)). The calorie of isomaltulose is 4kcal / g. In the present invention, isomaltulose includes palatinose syrup, reduced palatinose or palatinose syrup. Palatinose starch syrup is a starch syrup-like liquid substance mainly composed of oligosaccharides such as tetrasaccharide, hexasaccharide and octasaccharide produced by dehydration condensation of isomaltulose.

With respect to isomaltulose, it is known that a low glycemic food composition containing isomaltulose or the like is used for the treatment / prevention of malignant tumor diseases (Japanese Patent Publication No. 2009-530326).

The blending amount of isomaltulose is blended with other ingredients (milk protein hydrolyzate, fermented milk protein, fats and oils containing oleic acid, milk phospholipids, soybean lecithin, etc.) It can be adjusted as appropriate according to age, weight, use and the like. Specifically, the blending amount of isomaltulose can be exemplified by 4 to 15 g, preferably 5 to 7 g per 100 mL of the nutritional composition, but is not limited to these ranges (Special Table No. 2009-530326) Publication).

The amount of heat of the nutritional composition of the present invention can be adjusted by appropriately adding proteins, lipids and carbohydrates. The calorie | heat amount of the nutrition composition of this invention can illustrate 50-150 kcal per 100 ml of nutrition composition, Preferably, it is 80-120 kcal, However, It is not limited to these ranges.

In addition, the energy ratio of the protein, lipid, and carbohydrate to the whole nutritional composition in the nutritional composition of the present invention substantially conforms to the nutritional requirements of the sixth revised Japanese. Specific examples include 15 to 25% protein, 20 to 30% lipid, and 45 to 65% carbohydrate. However, the present invention is not limited to these ranges.

The nutritional composition of the present invention may also contain dietary fiber. Dietary fiber refers to substances in food that are not hydrolyzed by human digestive enzymes, and is classified into water-soluble dietary fiber and insoluble dietary fiber based on their affinity for water. As water-soluble dietary fiber, indigestible oligosaccharides such as lactulose, lactitol, or raffinose can be used. Physiological functions of indigestible oligosaccharides are known to reach the large intestine as undigested substances, contribute to the activation and proliferation of intestinal bifidobacteria, and have an improved intestinal environment, ie, an intestinal regulating effect. Yes.

Furthermore, as water-soluble dietary fiber candidates, high-molecular water-soluble dietary fiber includes soybean thickening polysaccharide, konjac glucomannan, alginic acid, low molecular alginic acid, psyllium, gum arabic, seaweed polysaccharide (cellulose, lignin-like substance, agar). , Carrageenan, alginic acid, fucodyne, laminarin), microbial gum (welan gum, curdlan, xanthan gum, gellan gum, dextran, pullulan, lambzan gum), other gums (seed-derived locust bean gum, tamarind gum, tara gum, sap-derived caraya gum, Polydextrose of low molecular weight water-soluble dietary fiber such as tragacanth gum), indigestible dextrin, pine fiber, maltitol and the like can be used.

Insoluble dietary fiber increases the mass of indigestibles in the large intestine and shortens transit time. As a result, the number of defecations increases, resulting in an increase in stool volume. Insoluble dietary fiber candidates include cellulose, hemicellulose, lignin, chitin, chitosan, soy bran, wheat bran, barley bran, pine fiber, corn fiber, beet fiber, oat bran, rye bran, pearl bran, rice bran, millet, millet Millet bran such as Japanese millet, sorghum, cereal bran, legume bran, sesame bran, and okara. Oat bran, rye bran, barley bran, rice bran, millet, millet, millet, sorghum bran, cereal bran, buckwheat bran, sesame bran, and okara.

The nutritional composition of the present invention includes water, protein, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors, emulsifiers in addition to the proteins, lipids, carbohydrates, and dietary fibers. , Thickeners, stabilizers and the like can be used. Examples of the protein include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, whey protein hydrolysate, α-casein, β-casein, κ- Casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and vegetable proteins, their degradation products; butter, whey minerals, cream, whey, non-protein nitrogen And various milk-derived components such as sialic acid, phospholipid, and lactose. It may contain peptides such as casein phosphopeptides and lysines and amino acids. Examples of the saccharide include saccharides, processed starch (in addition to text phosphorus, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of lipids include animal oils such as lard and fish oil, fractionated oils thereof, hydrogenated oils and transesterified oils; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, tartaric acid, erythorbic acid, and the like. In addition, a material containing a fecal odor reducing effect (for example, champignon extract 5 mg to 500 mg (0.005% to 0.5%)), a carotenoid preparation (for example, α-carotene, β-carotene, lycopene, lutein, etc.) 10 μg to 200 μg (0.00001% to 0.0002%)) and an antioxidant (catechin, polyphenol, etc.) can also be included. Two or more of these components can be used in combination, and a synthetic product and / or a food containing a large amount thereof may be used. The form of the food may be any form such as a solid, liquid, or gel.

  The nutritional composition of the present invention can be produced by methods known in the art. After some or all of the raw materials have been prepared, homogenization is performed as necessary. Homogenization means homogenization by thoroughly mixing each prepared component, and the fat globules and coarse particles of other components are mechanically refined to prevent the rising and aggregation of fat and the like, and the nutritional composition It means making into a uniform emulsified state.

In the production of the nutritional composition of the present invention, heat treatment or heat sterilization is performed. As heat sterilization conditions, general food sterilization conditions can be used, and heat sterilization can be performed using a conventional apparatus. For example, sterilization of 62-65 ° C. × 30 minutes, 72 ° C. or more × 15 seconds or more, 72 ° C. or more × 15 minutes or more, or 120-150 ° C. × 1-5 seconds, or 121-124 ° C. × 5-20 minutes, 105 Although sterilization of ˜140 ° C., retort (pressure heating) sterilization, high-pressure steam sterilization, and the like can be used, it is not limited to these examples. The heat sterilization can be preferably performed under pressure.

In addition, after preliminarily heat-sterilizing the liquid nutritional composition, aseptically filling the container (for example, a method using a combination of the UHT sterilization method and the aseptic packaging method), filling the container with the liquid nutritional composition, Method of heat sterilization with containers (for example, autoclave method), can containers, liquid foods, various containers used for oral and tube feeding (so-called soft bags, nutrition bags, etc.) and retort sterilization (for example, at 115 to 145 ° C 5 ~ 10 seconds), filling in cans, liquid foods and various containers used for oral and tube feeding (so-called soft bags, nutrition bags, etc.) and sterilized by retort for about 5-8 seconds at about 140-145 ° C Although the method of cooling after heat sterilization and performing aseptic filling can be illustrated, it is not limited to these examples.

By the heat treatment or heat sterilization, starters (such as lactic acid bacteria, bifidobacteria, or Propionibacterium spp.) Derived from fermented milk as a raw material are killed.

The composition of the present invention can be used to suppress the growth of cancer. The nutritional composition of the present invention can be used for nutritional management of oral and enteral nutrition patients, the elderly, infants and the like, for example, as liquid foods, oral and tube feedings, beverages, gel foods and the like.

The osmotic pressure of the nutritional composition of the present invention can be exemplified by an osmotic pressure of about 500 to 1000 mOsm / l, such as about 550 to 750 mOsm / l. When measured at room temperature, the viscosity of the nutritional composition can be exemplified by 20-100 cp (1 cp = 1 mPa · s), preferably 25-60 cp, more preferably 30-50 cp. The range is not limited.
The pH of the nutritional composition of the present invention can be adjusted to 4.6 or less, preferably 3.0 to 4.3, more preferably 3.8 to 4.2, but is not limited to these ranges.

The nutritional composition of the present invention is beneficial in inhibiting tumor growth in tumor patients. In particular, colon tumor patients, colon solid tumor patients, sarcoma cell tumor patients, lung cancer patients, thymoma patients, breast cancer patients, pharyngeal cancer patients, laryngeal cancer patients, gastric cancer patients, esophageal cancer patients, hepatocellular carcinoma patients, cholangiocarcinoma patients, Gallbladder cancer patients, pancreatic cancer patients, penile cancer patients, renal pelvis and ureter cancer patients, renal cell cancer patients, testicular cancer patients, prostate cancer patients, bladder cancer patients, vulvar cancer patients, cervical cancer patients, endometrial cancer patients It is useful for inhibiting the growth of tumors such as uterine sarcoma patients, vaginal cancer patients, ovarian cancer patients, ovarian germ cell tumor patients, or brain tumor patients.

In an in vivo test using a mouse transplanted with a tumor cell line, it was confirmed that the nutritional composition of the present invention inhibits tumor growth and tumor infiltration into surrounding tissues. When tumors invade surrounding tissues, tumor metastasis is likely to occur. In addition, pain occurs when a tumor infiltrates an internal organ or peripheral nerve. Therefore, the composition of the present invention can suppress tumor growth, tumor invasion / metastasis, or tumor-induced pain.

The daily intake of the nutritional composition of the present invention in a pharmaceutical product or food / drink depends on the pathology, age, symptoms, weight, use of the subject, and whether the nutritional composition of the present invention is the only nutritional item. Since it differs, it is not specifically limited. Specifically, in the case of an adult, the intake per day may be 400 to 1600 ml, preferably 600 to 1600 ml, more preferably 800 to 1200 ml. For adults, for example, up to 3000 ml per day may be taken. The intake amount can also be determined by the attending physician of the subject person.
Moreover, you may use the nutrition composition of this invention together with the pharmaceutical and foodstuff which have an antitumor action conventionally known.

The nutritional composition of the present invention can be used in the form of any medicine or food or drink. For example, the intestinal flora can be obtained by directly administering the nutritional composition of the present invention as a pharmaceutical or by directly ingesting it as a special-purpose food such as a food for specified health use, a nutritional functional food, a nutritional supplement, a liquid food or a supplement. Is expected to improve. In addition, various foods (milk, soft drinks, fermented milk, yogurt, cheese, bread, biscuits, crackers, pizza crusts, prepared milk powder, liquid foods, special-purpose foods, regardless of the form of liquid, paste, solid, powder, etc. , Foods for the sick, nutritional foods, frozen foods, processed foods, and other commercially available foods). Moreover, when the usage form of a nutrient composition is a powder, it can manufacture by using means, such as spray drying and freeze-drying, for example.

When the nutritional composition of the present invention is used as a pharmaceutical or a supplement, it can be administered in various forms. Examples of the form include oral administration using tablets, capsules, granules, powders, syrups and the like. These various preparations can be generally used in the pharmaceutical preparation technical field such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizers, suspension agents, coating agents, etc. as main ingredients according to conventional methods. It can be formulated using known adjuvants for formulation. Further, it may contain an appropriate amount of calcium. Further, an appropriate amount of vitamins, minerals, organic acids, sugars, amino acids, peptides, etc. may be added.
Example

Hereinafter, although an example is given and explained about the present invention, the present invention is not limited by this.
Materials and methods

Two commonly used cancer cell models, mouse sarcoma cells (S180) and mouse colon solid tumor cells (C26), construct S-180 as an ascites tumor model and C-26 as a solid tumor model. Randomly divide the mice into two groups so that half are male and half are female. The group fed with the nutritional composition of the present invention with 14 days of constant feeding is called “test composition of the present invention” and AIN-93M fed with the control diet. The survival time of mice bearing S-180 tumors was recorded as the main index, and food intake, body weight, tumor weight, and tumor volume of mice bearing C-26 tumors were recorded. Serum TNF-a and IL-6 levels were measured and tumor histopathology was also performed.
1. Meal test

  The test composition (yellow fine powder) of the present invention is a dried liquid food composition shown in Tables 1 and 3. The total fatty acid composition is formed with 39% by weight of liquid oleic acid, 1.2% by weight of eicosapentaenoic acid and 0.8% by weight of docosahexaenoic acid, and AIN-93M (white fine powder) is manufactured by Oriental Yeast Industry (Japan). A diet refined to the AIN-93M standard, commonly used in nutrition experiments in rats and mice, and its ingredients were published in 1993 by the United States National Institute of Nutrition (Table 4). And Table 5).

2. Transplanted tumor cell model
S-180 cells were provided by the Shanghai Institute of Oncology.
C-26 cells were provided by the Shanghai Institute of Oncology.
3. animal

Variety: BALB / c mice
Source: Shanghai Shanghai SIPPR-BK Laboratory Animal Co. Ltd
Gender: Half male and half female Age: 6-7 weeks Weight: 18-20g
Number: 76 (36 mice for S-180 experiments, 40 mice for C26 experiments)
Animal serial number: SCXK (Shanghai) 2008-0016
Animal license number: SYXK (Shanghai) 2008-0027
Storage conditions: SPF Animal Experiment House, room temperature 24 + 1 ° C, humidity 40-70%, 12/12 hours and night cycle.
4). Method

Collect S-180 cell line and C26 cell line one week before inoculation. RALB / c mice are used to create a murine tumor model. The mouse is grabbed and the skin is disinfected, and S-180 cells are inoculated into the abdominal cavity of the mouse, or C26 cells are inoculated subcutaneously near the right rear axilla of the mouse. On the day of inoculation, animals were randomly divided into AIN-93M group and test composition group and placed separately in cages.

36 mice (18 per group) were used for S-180 ascites tumor studies. Six mice were bred in each cage with two special food feeders, each filled with about 40 g of the diet. The mice were given food freely. Food intake was weighed every other day, body weight was recorded every 3 days, and the number of dead mice was recorded.

Forty mice (20 in each group) were used for the C26 solid tumor study. Five mice were bred in each cage with two special food feeders, each filled with about 40 g of the diet. The mice were given food freely. Tumor volume and food intake were weighed every other day and body weight was recorded every 3 days. On the 14th day after inoculation, the mouse was sacrificed by cervical dislocation and eye blood was collected. Tumors were dissected and removed, tumor volume was measured, tumor weight was also measured, and the tumor tissue was placed in 10% formalin for histopathological examination.

Tumor volume calculation formula: V (cm ³ ) = a * b2 / 2 (a = major axis (cm), b = minor axis (cm))
Tumor growth inhibition rate (%) = (mean tumor volume of control experiment group−mean tumor volume of test composition group) × 100% / mean tumor volume of control experiment group, or
Tumor growth inhibition rate (%) = (average tumor weight of control experiment group−average tumor weight of test composition group) × 100% / average tumor weight of control experiment group

Drug efficacy criteria: Assume that the difference between the mean tumor weight (or tumor volume) of the two groups is statistically significant in the situation of P <0.05.
5. Statistical analysis

Data was analyzed using statistical software SPSS version 11.5. In the study of S-180 tumors, the mean survival time and cumulative survival rate of the two groups were compared with the survival analysis log rank test. In the C26 tumor study, the average tumor weight and average tumor volume of the two groups were compared using Student's Ty test. P <0.05 was considered statistically significant.
III Results 1. S-180 ascites tumor study in mice

The average survival of mice in the test composition group was 13.0 ± 0.3 days (mean ± SE), and the average survival of mice in the AIN-93M group was 12.4 ± 0.4 days. There was no significant difference between the two groups with respect to either mean survival days or cumulative survival (P> 0.05 in the logarithmic rank test for survival analysis) (Table 6, Table 7). Although there were no significant differences, the survival curve revealed the following: That is, the survival rate of the mice in the test composition group was higher than that of the control experimental group, and the two survival curves did not intersect at any time (Fig.1, Fig.2). This means that the mouse survival time of the test composition group tended to be longer than that of the control experimental group.
2. Murine C26 solid tumor study 2.1 Food intake and body weight

During the entire experiment, the average food intake of the tumor-bearing mice in the test composition group was 2.60 ± 0.49 grams per day for each mouse, and the average food intake of the tumor-bearing mice in the control group was per day for each mouse. 3.37 ± 0.45 grams (Table 8). There was no significant difference in food intake between the two groups (P> 0.05) (Fig. 2). There was no significant difference between the two groups in weight change (P> 0.05) (Fig. 3).
2.2 Tumor weight and tumor volume

As shown in Table 8, the average tumor weight of the mice in the test composition group was 1.36 ± 0.54 grams, and the average tumor weight of the control test group was 1.83 ± 0.89 grams, indicating a significant difference (P <0.05). ). The tumor growth inhibition rate of the test composition group calculated by tumor weight was 25.59%.

Similarly, the average tumor volume for mice in the test composition group was 1.868 ± 0.84 grams and the average tumor volume for the control test group was 2.582 ± 1.055 grams. The mean tumor volumes measured on days 4, 7 and 11 were all significantly greater in the test composition group than in the control test group (P <0.05). The tumor growth inhibition rate of the test composition group calculated by tumor volume was 27.65% (Table 9, Fig. 4, Fig. 5).
2.3 Tumor pathology

Upon tumor tissue dissection, the tumor integrity in the test composition group was better, had a clear framing and was found to be easier to detach. In contrast, tumors in the ALN-93M group appeared to have invaded surrounding tissues and were difficult to detach.

Histopathological examination revealed the following. That is, the outline of the tumor in the test composition group was smooth, but cancer cells and hemorrhagic lesions were distributed in the marginal area, and darkly colored cells grew densely in the central area (FIG. 6A). These cancer cells had various sizes and shapes with darkly colored nuclei (FIG. 6B). Tumor outlines in the AIN-93M control study group were unclear, bleeding was seen in a wide area (FIG. 6C), cancer cells were spread, necrosis was evident and surrounded by bleeding (FIG. 6D).

A pathology study noted the following: That is, malignant tumors in the test composition group were less invasive than those in the control test group. This suggests that the test composition has some degree of deterrent effect on the tumor.

A. Test composition group: The tumor profile in the test composition group is clean and smooth, the cancer cells are scattered, and a homogeneous eosin-like substance (ie bleeding) is seen. Darkly colored cancer cells are found in the central region.

B. Test composition group: Cancer cells grow densely in the central area, cells are of various sizes and shapes, and homogeneous eosin-like material (ie bleeding) is found in several narrow areas. (Upper right, high magnification: Cancer cells and nuclei of various shapes and sizes were heavily colored. HEx400)

C. Control study group: The tumor margin is not uniform and there is a wide area of homogeneous eosin-like substance (ie bleeding).

D. Control study group: Cancer cells spread, necrotic lesions visible and surrounded by homogeneous eosin-like substance (ie bleeding).

As a commonly used mouse cancer model, ascites tumor model S-180 and solid tumor model C-26 are used to determine whether the test composition of the present invention can inhibit tumor growth. evaluate. As a result, in the test set of the present invention, the mean survival time of the mice having S-180 tumor was not significantly different from the control mice (P> O. 05). However, the test set of the present invention was able to inhibit tumor growth in the C-26 mouse tumor model.

The nutritional composition of the present invention is beneficial for colon solid tumor patients by inhibiting colon solid tumor growth or reducing malignant tumor invasion.

Claims (16)

Nutritional composition suitable for tumor patients comprising hydrolyzate of milk protein as protein and fermented milk protein, fat containing oleic acid as lipid, milk phospholipid and / or soy lecithin, isomaltulose as carbohydrate . The milk protein is selected from the group consisting of casein, milk protein concentrate (MPC), whey protein concentrate (WPC), whey protein isolate (WPI), α-lactalbumin, β-lactoglobulin and lactoferrin. The nutritional composition according to claim 1, wherein: The nutritional composition according to claim 1, wherein 0.9 to 5.0 g of milk protein hydrolyzate is contained per 100 ml of the nutritional composition. The milk protein hydrolyzate may be obtained by hydrolyzing a whey protein isolate (WPI) with an alcalase derived from Bacillus licheniformus and with a trypsin derived from porcine pancreas. The nutritional composition according to claim 1. The nutritional composition according to claim 4, wherein the milk protein hydrolyzate is a permeate obtained by treatment with an ultrafiltration membrane having a fractional molecular weight of 10,000. The nutritional composition according to claim 1, wherein the fermented milk protein is derived from cheese. The nutritional composition according to claim 6, wherein the cheese is quark. The nutritional composition according to claim 1, wherein 0.5-6g of the fermented milk protein is contained per 100ml of the nutritional composition. The nutritional composition according to claim 1, wherein 4-15g of isomaltulose is contained per 100ml of the nutritional composition. The nutritional composition according to claim 1, wherein the lipid contains oleic acid at 30% or more of the total fatty acid composition.   Nutritional composition suitable for tumor patients, including milk protein hydrolyzate and fermented milk protein fat as protein, fat containing oleic acid as quality, milk phospholipid and / or soy lecithin, isomaltulose as sugar A method for inhibiting tumor growth comprising the step of administering an agent. 12. The method of claim 11, wherein the tumor patient is a colon solid tumor patient. 12. The method according to claim 11, wherein the nutritional composition is an oral or tube food intake (enteral) nutritional composition.   Nutritional composition suitable for tumor patients, including milk protein hydrolyzate and fermented milk protein fat as protein, fat containing oleic acid as quality, milk phospholipid and / or soy lecithin, isomaltulose as sugar A method of treating or shrinking a tumor, comprising administering an agent.   15. The method of claim 14, wherein the tumor is a colon solid tumor.   15. The method according to claim 14, wherein the nutritional composition is an oral nutritional agent.