NZ613689B2 - Agent for improving deteriorated peripheral sensation due to aging and/or disease - Google Patents

Abstract

Disclosed is a use of a milk-derived basic protein fraction in the manufacture of a medicament for improving deteriorated peripheral sensation due to ageing and/or a disease.

Description

DESCRIPTION AGENT FOR IMPROVING DETERIORATED PERIPHERAL SENSATION DUE TO AGING AND/OR DISEASE TECHNICAL FIELD The invention relates to a skin sensitivity improving agent that includes a basic protein fraction or a ed basic protein fraction as an active ingredient and improves deterioration in peripheral sensation, and a skin sensitivity improving food, drink, feed, or cosmetic that includes the skin sensitivity improving agent.

BACKGROUND ART In recent years, an increase in age-related diseases such as osteoporosis, dementia and the like has become a serious social issue along with aging of population.

Various drugs have been developed to prevent or improve such age-related diseases. r, since drugs always need to take side effects into consideration, in recent years, attempts have been made to prevent or improve age-related diseases through a change in eating habits or intake of a specific food ingredient. For example, it is known that osteoporosis is prevented or improved by intake of a basic protein contained in cow milk (see Patent Document 1). A dementia therapeutic agent that prevents or improves Alzheimer-type ia and contains sphingomyelin which is one of the phospholipids containing relatively ntly in cow milk as an active ingredient has also been known (see Patent Document 2). oration in eral ion can be known as one of the lated symptoms. The deterioration in peripheral sensation also occurs due to not only aging, but also diseases such as diabetes and the like. Deterioration in peripheral sensation may occur following troubles; for example, as it can’t feel hot immediately when touching a hot object, a risk of burns or the like increases or the discovery of an injury becomes delay due to deterioration in pain sensation. In recent year, in order to decrease such a risk, s that improve deterioration in peripheral sensation due to ageing or diseases have been conducted. For example, it has been reported that sphingomyelinase or phosphatidylcholine-specifi'c phospholipase C, which are enzymes that increase biosynthesis 0f endogenous ceramides promote differentiation of PC-12 cells which is one of the neural cell lines through the secretion of neurotrophic factors from Swiss 3T3 fibroblast cells (see Non-patent nt 1). r, since ceramides and the above enzymes are not food ingredients, it is necessary to take account of safety. Therefore, a safe agent that is effective to improve deterioration in peripheral sensation h daily intake or skin application has been desired.

PRIORART DOCUMENT PATENT DOCUMENT Patent Document 1: JP-A-H08-1513 31 Patent Document 2: JP-A146883 TENT DOCUMENT Non-patent Document 1: Norimichi ta and Satoko Okubo, s on Expression Mechanism of Physiological Functions of Ceramides Having Skin Protection Function, Annual Report of Cosmetology, Vol. 10, 2002 SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION In some embodiments the present invention seeks to provide a safe and skin sensitivity improving agent that is effective to improve deterioration in eral sensation through daily intake or skin application. In other embodiments the present invention seeks to provide a sensation-improving food, drink, feed, or cosmetic that is effective to improve deterioration in peripheral sensation through oral intake or skin application.

MEANS FOR SOLVING THE PROBLEMS The inventors of the invention, in consideration of those problems, searched for a safe component that ts an excellent improvement effect to the deterioration in sensation. As a result, the inventors found that deterioration in sensation, particularly peripheral ion, can be improved by oral intake or skin application of a basic n fraction derived from milk (hereinafter referred to as “milk-derived basic protein fraction”) or a degraded basic protein fraction obtained by ing the milk-derived basic protein fraction using a protease (hereinafter referred to as “degraded milk-derived basic protein fraction”). The ors thus completed a skin ivity improving agent by utilizing the milk-derived basic protein fraction and/or the degraded milk-derived basic protein fraction as an active ingredient. The ors also found that a skin sensitivity ing food, drink, feed, or cosmetic can be obtained by adding the skin sensitivity improving agent to a food, drink, feed, or the like, respectively. These findings have led to completion of the invention. ically, the invention es the following. (1) A skin sensitivity improving agent including a milk-derived basic protein fraction as an active ingredient. (2) The skin sensitivity improving agent according to (1), wherein the milk-derived basic protein fraction contains basic amino acids in an amount of 15 wt% or more based on total amino acids. (3) The skin sensitivity improving agent according to (1) or (2), wherein the milk-derived basic protein fraction is obtained by bringing milk or a milk-derived raw material into contact with a cation-exchange resin to allow basic proteins to be adhered on the cation-exchange resin, and eluting the fraction adhered on the cation-exchange resin using an eluant having a sodium chloride tration of 0.1 to 1.0 M. (4) A skin sensitivity ing agent including a degraded milk-derived basic protein fraction as an active ingredient, the degraded milk-derived basic protein on being obtained by treating the milk-derived basic protein fraction according to any one of (1) to (3) with a protease. (5) The skin sensitivity improving agent according to (4), n the protease is at least one protease selected from the group consisting ofpepsin, trypsin, chymotrypsin, and pancreatin. (6) A skin sensitivity improving food, drink, feed, or cosmetic including the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction ing to any one of (1) to (5). (7) A method for ing sensation of a mammal including administering a composition that includes a milk-derived basic protein fraction to the mammal via oral administration or application on skin. (8) The method according to (7), wherein the milk-derived basic protein fraction contains basic amino acids in an amount of 15 wt% or more based on total amino acids. (9) The method according to (7) or (8), wherein the milk-derived basic protein on is obtained by ng milk or a milk-derived raw al into t with a cation-exchange resin to allow basic proteins to be adhered on the cation-exchange resin, and eluting the fraction adhered on the cation-exchange resin using an eluant having a sodium chloride concentration of 0.1 to 1.0 M. (10) A method for improving sensation of a mammal ing administering a composition that includes a degraded milk-derived basic protein fraction to the mammal via oral administration or application on skin, the degraded milk-derived basic protein fraction being obtained by treating the milk-derived basic protein fraction according to any one of (7) to (9) with a protease. (11) The method according to (10), wherein the protease is at least one protease selected from the group consisting of , n, chymotrypsin, and pancreatin.

EFFECTS OF THE ION [0008a] In a first aspect the present invention provides a use of a milk-derived basic protein fraction in the manufacture of a medicament for improving deteriorated peripheral sensation due to ageing and/or a disease.

In a second aspect the present invention es a use of a degraded milk-derived basic protein fraction in the manufacture of a medicament for improving deteriorated peripheral sensation due to ageing and/or a disease, wherein the degraded milk-derived basic protein fraction is obtained by treating a milk-derived basic protein fraction with a protease. [0008c] In a third aspect the present invention provides a use of a milk-derived basic protein on in the preparation of a food, drink, feed, or cosmetic for improving deteriorated peripheral sensation due to ageing and/or a disease. [0008d] In a fourth aspect the present invention provides a use of a degraded milk-derived basic protein fraction in the preparation of a food, drink, feed, or cosmetic for improving orated peripheral sensation due to ageing and/or a disease, n the degraded milk-derived basic n fraction is obtained by treating a milk-derived basic protein fraction with a protease.

The ion can thus provide a skin sensitivity improving agent that includes a milk-derived basic n fraction or a degraded milk-derived basic protein fraction as an active ingredient, and a skin sensitivity improving food, drink, feed, or cosmetic that includes a milk-derived basic protein fraction or a degraded milk-derived basic protein fraction. The skin sensitivity improving agent according to the invention exhibits an effect that improves deterioration in peripheral sensation.

EMBODIMENTS FOR CARRYING OUT THE INVENTION The invention is characterized in that a milk-derived basic protein fraction or a degraded milk-derived basic protein on are used as active ingredients. The milk-derived basic protein fraction is ed from mammalian milk such as cow milk, human milk, goat milk, or ewe milk, and the degraded milk-derived basic protein fraction is obtained by treating the milk-derived basic protein fraction with a protease.

The milk-derived basic protein fraction has the ing properties. 1) The milk-derived basic n fraction comprises several types of proteins having a molecular weight of 3,000 to 80,000 determined by sodium dodecyl sulfate rylamide gel ophoresis (SDS-PAGE). 2) The milk-derived basic protein fraction includes proteins in an amount of 95 or less by treating a milk-derived basic protein fraction obtained by the above methods with a protease such as pepsin, trypsin, or chymotrypsin, and optionally treating the resulting product with a protease such as pancreatin or the like.

The milk-derived basic protein fraction Or the degraded erived basic protein fraction may be used ly as the skin sensitivity improving agent according to the invention. Note that the skin sensitivity improving agent may be further mixed a raw material or the like that is normally used for drugs, food, drink, and feed, such as saccharides, lipids, ns, vitamins, minerals, or , or the like, in on to the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction, and the skin sensitivity improving agent may be prepared into a powdered drug, granules, a tablet, a capsule, a drinkable preparation, or the like by a tional method. The skin sensitivity improving agent may also be used in common application form such as emulsion, cream, lotion, massage mask, or the like. The skin sensitivity improving agent in preparation form may be prepared by a conventional method while appropriately adding the milk-derived basic protein fraction or the degraded milk-derived basic protein on used as an active ingredient in the invention, and may be used as a cosmetic. Another component, e.g., ceramide, sphingomyelinase, or sphingomyelin or the like that exhibits a skin sensitivity ing effect may be used in combination with the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction. In the experiments described later using mice, the peripheral sensation was improved by orally administering the milk-derived basic protein fraction or the degraded erived basic protein fraction in an amount of 10 mg/kg weight or more, and preferably 20 mg/kg weight or more per mouse. Therefore, deterioration in sensation, particularly peripheral sensation, is expected to be ed when an adult lly takes the milk-derived basic protein fraction or the degraded erived basic protein fraction in an amount of 10 mg/day or more, and preferably 20 mg/day or more, and thus it is desired to take the above necessary quantity. When applying the milk-derived basic protein fraction or the degraded milk-derived basic protein on to skin, the skin liniment may contain the milk-derived basic protein on or the degraded milk-derived basic protein fraction in an amount of 0.001 to 40 wt%, and preferably 0.1 to 10 wt%, based on the total amount of the skin nt.

A skin sensitivity improving food or drink according to the invention may be produced by mixing the milk-derived basic protein fraction or the degraded milk-derived basic protein on to a normal food or drink, such as yogurt, milk-based drink, wafer, dessert or the like. It is preferable that the skin ivity improving food or drink contain the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction in an amount of 0.5 to 2000 mg per 100 g of the food or drink so that an adult can take the erived basic protein fraction or the degraded milk-derived basic n fraction in an amount of 10 mg/day or more, although the content of the milk-derived basic protein on or the ed milk-derived basic protein fraction is appropriately determined depending on the type of the food or drink. A skin ivity improving feed according to the invention may be produced by adding the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction to a normal feed, such as livestock feed, pet food or the like. It is preferable that the skin sensitivity improving feed contain the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction in an amount of 0.5 to 2000 mg per 100 g of the feed so that the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction is taken in an amount of 10 mg/day or more.

The method of mixing the milk-derived basic protein fraction or the degraded erived basic protein fraction may not be particularly d. For example, the milk-derived basic protein fraction or the ed erived basic protein fraction is suspended or ved in deionized water, and the mixture is stirred and prepared in the form of a drug, food, drink, or feed. The stirring/mixing conditions are not particularly d as long as the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction is unifome mixed. The mixture may be stirred/mixed using an ultra-disperser, a TK-homomixer, or the like. The solution containing the composition may optionally be concentrated using an RO membrane, or freeze-dried so that the solution can be easily used for a drug, food, drink, or feed. A sterilization treatment conventionally used in the production of a drug, food, drink, or feed may be employed in the invention. at sterilization may also be employed for a powdery product. Therefore, it is possible to e drugs, food, drink, and feed in various forms (e.g., liquid, gel, powder, or granules) that contain the milk-derived basic protein fraction or the degraded milk-derived basic protein fraction ofthe invention.

The invention is further described below by way of examples and test examples.

Note that the following examples are for illustrative purposes only, and should not be construed as limiting the invention.

Example 1 A column (diameter: 5 cm, height: 30 cm) filled with 400 g of sulfonated Chitopearl (cation-exchange resin; manufactured by Fuji ng Co., Ltd.) was sufficiently washed with deionized water. 40 1 (liters) of unsterilized skim milk (pH 6.7) was passed through the column at a flow rate of 25 ml/min. The column was then sufficiently washed with zed water, and a basic protein fraction adhered on the resin was eluted with a 0.02 M carbonate buffer solution (pH 7.0) containing 0.98 M sodium chloride. The eluate was desalted and trated using a reverse s (RO) membrane, and freeze-dried to obtain 21 g ofpowdery milk-derived basic protein fraction (Example product 1). The molecular weight of the milk-derived basic protein fraction determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis AGE) was distributed in the range of 3000 to 80,000. The milk-derived basic protein fraction had the composition shown in Table 1. The milk-derived basic protein fraction was hydrolyzed using 6 N hydrochloric acid at 110°C for 24 hours, and the amino acid composition of the milk-derived basic protein fraction was analyzed using an amino acid analyzer (“L-8500” manufactured by i Ltd.). The results are shown in Table 2. The protein composition ofthe milk-derived basic protein fraction was analyzed by ELISA. As shown in Table 3, the milk-derived basic protein on had a lactoferrin content and a lactoperoxidase content of 40% or more. The basic protein fraction may be used directly as the skin ivity improving agent according to the invention.

TABLE 1 sh .

TABLE 2 Aspartic acid 10.1 (wt%) Arginine .

TABLE 3 Example 2 A column (diameter: 100 cm, height: 10 cm) filled with 30 kg of cation-exchange resin (SP Toyopearl; manufactured by Tosoh Corporation) was sufficiently washed with deionized water. 3 t of cheese whey (pH 6.2) that had been heat-sterilized at 121°C for 30 seconds was passed through the column at a flow rate of l/min. The column was then sufficiently washed with deionized water, and a basic n fraction adhered on the resin was eluted with a 0.1 M citrate buffer on (pH .7) containing 0.9 M sodium chloride. The eluate was desalted and concentrated by electrodialysis (ED), and freeze-dried to obtain 183 g of a powdery milk-derived basic protein on (Example product 2). The milk-derived basic protein fraction thus ed may be used directly as the skin sensitivity improving agent according to the invention.

Example 3 50 g of the milk-derived basic protein on obtained in Example 1 was dissolved in 10 l of led water. After the addition of 1% atin (manufactured by Sigma), the mixture was reacted at 37°C for 2 hours. After completion ofthe reaction, the protease was inactivated by heating the mixture at 80°C for 10 minutes to obtain 48.3 g of a degraded milk—derived basic protein fraction (Example product 3).

The degraded milk—derived basic n fraction thus obtained may be used directly as the skin sensitivity improving agent according to the invention.

Test Example 1 (Study on cell differentiation—promoting activity) Swiss 3T3 cells, which is one of the fibroblast cell lines known to be present in skin, were cultured for 2 days under the condition that Example product 1 of the milk-derived basic n fraction or Example product 3 of the degraded milk—derived basic protein fraction was added at a concentration of 0.03 to 1%, respectively (Example t 1: group A, e product 3: group B). As a control, Swiss 3T3 cells were cultured for 2 days without adding Example product 1 of the milk—derived basic protein fraction or Example product 3 of the degraded milk—derived basic protein fraction (group C). PC—12 cells, which is one of the neural cell lines, were cultured adding the culture medium of Swiss 3T3 cells, and morphological differentiation of the PC-12 cells was observed.

As the results, PC-12 cells were highly differentiated when adding the culture mediums ofthe group A or B. The above experiment was repeated several times, and the difl'erentiation ratio was determined using an optical cope. It was confirmed that 95% or more ofthe cells were differentiated in both cases of group A and B. ' contrast, PC-12 cells were not differentiated when adding the culture medium of the group C. No differentiation was observed using an l microscope when the experiment was repeated several times. It was thus confirmed that the milk-derived basic protein fraction obtained in Example 1 and the ed milk-derived basic protein fraction obtained in Example 3 promoteddifferentiation of PC-12 cells which is one ofthe neural cell lines.

Test Example 2 (Study on skin sensitivity improving effect using experimental animals) A skin ivity improving effect was evaluated by the hot plate test that is a thermal stimulation behavioristic approach developed by Woolfe and MacDonald. 24-week-old hairless mice R-l) were divided into three groups (10 mice/group).

Example product 2 (milk-derived basic protein fraction) was orally stered to each mouse using a sonde in an amount of 0, 10, or 20 mg/kg weight once daily for 4 weeks.

Each mouse was placed on a hot plate at 54°C, and the time elapsed until the mouse made an escape behavior such as taking off the foot from the hot plate, standing up, or jumping was measured. The m time of escape behavior positive reaction time that is time until the mouse made an escape behavior after applying l stimulation was set to 30 seconds. When the escape behavior positive reaction time was 30 seconds or more, the escape behavior positive reaction time was determined to be 30 seconds. The results are shown in Table 4.

TABLE 4 Escape behavior 29.2.0443... momec .1 iozssec As shown in Table 4, the escape behavior positive reaction time tended to be shortened when administering Example product 2 (milk-derived basic protein fraction) in an amount of 10 mg, and significantly shortened when administering Example product 2 (milk-derived basic n fraction) in an amount of 20 mg. It was thus confirmed that deterioration in sensation, particularly peripheral ion, can be prevented or improved by intake of Example product 2 derived basic protein on).

Test Example 3 (Study on skin sensitivity improving effect by oral intake) Healthy elderly persons (average age: 752t3) who suffered deterioration in sensation in the hands were divided into following five groups (10 subjects/group).

Group A: the subjects did not take a milk-derived basic protein fraction or a milk-derived basic protein fraction, Group B: the subjects took the milk-derived basic protein fraction obtained in Example 1 in an amount of 10 mg for 6 weeks, Group C: the subjects took the milk-derived basic protein fraction obtained in e 1 in an amount of 20 mg for 6 weeks, Group D: the subjects took the degraded erived basic protein fraction obtained in Example 3 in an amount of 10 mg for 6 weeks, and Group E: the subjects took the degraded milk-derived basic protein fraction ed in Example 3 in an amount of 20 mg for 6 weeks.

The pain sensation in the palm and the arch of the foot was measured with 4 criteria (normal, deterioration I, deterioration II, and deterioration III) before the intake and after the intake for 6 weeks with nce to the pain sensation in the medial side of the arm using a pain/touch-pressure sensation measuring device (Algesiometer; manufactured by Intercross Ltd.) in accordance with the instruction manual. A questionnaire survey on an improvement in sensation in the hands was carried out to each elderly person after the intake for 6 weeks. The results are shown in Tables 5 to (Measuring method) The pain ion was evaluated using five pins having different thickness at five fiilcrum positions. The thinnest pin 1 was rolled along the medial side ofthe arm, and the subject was asked about the degree of normal pain sensation.

The pin 1 was then rolled along the palm and the bottom ofthe foot while sequentially changing the holder fillCI'llIIl on to determine the fillch position at which the same pain sensation degree as the first pain sensation occurred.

(Evaluation ) The algesiometer is ed so that the pain sensation occurs to the same extent between when rolling the pin 1 (fiilcrum: 50 g) along the medial side ofthe arm and when rolling the pin 2 (fillcrum: 50 g) along the palm. The pain sensation was evaluated as follows in accordance with the instruction manual. The pain sensation was evaluated by , and the average points were calculated.

Normal (0 points): The same pain sensation ed when rolling the pin 2 (fiilcrum: 50 g). oration I (1 point): The same pain sensation occurred when rolling the pin 1 (15110111111: 50 g).

Deterioration II (2 point): The same pain sensation occurred when rolling the pin 1 (fiilcrum: 60 g).

Deterioration III (3 point): The same pain sensation occurred when rolling the pin 1 (fiilcrum: 70 g).

TABLE 5 Hand sensation measurement (before intake) ---Deterioration Deterioration Deterioration Average s .

TABLE 6 Foot bottom sensation ement (before intake) ‘- - l- _— l- —— -n .- - ——_——_ TABLE 7 Hand sensation TABLE 8 Foot bottom sensation As shown in Tables 5 to 8, the sensation in the palm and the bottom of the foot tended to be improved by intake of Example product 1 derived basic protein fraction) or Example product 3 (degraded milk-derived basic protein fraction) in an amount of 10 mg, and was significantly improved by intake of e product 1 (milk-derived basic protein fraction) or Example product 3 (degraded milk-derived basic protein fraction) in an amount of 20 mg. Deterioration in sensation, particularly in peripheral sensation, is ed to be improved when an adult takes the milk-derived basic protein fraction or the ed milk-derived basic protein fraction in an amount of 10 mg/day or more generally, and preferably 20 mg/day or more.

Example 4 (Preparation of skin sensitivity improving cosmetic (cream)) A skin sensitivity improving cosmetic (cream) was prepared by mixing the ed milk-derived basic protein fraction (Example product 3) obtained in Example 3 with the raw materials in the ratio shown in Table 9. [003 5] TABLE 9 Glycerol monostearate (self-emulsifiable) Purified lanolin Liquid paraffin 50 Jojoba oil 5.0 Parabene 0.3 Degraded milk-derived basic protein fraction (Example production 3)_ ' Test Example 4 (Study on skin sensitivity improving effect via skin application) Healthy y persons (average age: 75:l:3) who suffered deterioration in ion in the hands were divided into groups A and B (15 subjects/group). The subjects of the group A were d the cosmetic (cream) that was prepared in the same manner as Example product 4, but did not contain a skin sensitivity improving agent, once daily to whole of the hands and the feet thereof, and the subjects of the group B were d the skin sensitivity improving cosmetic (cream) obtained in Example 4 once daily to whole of the hands and the feet f. The application period was 6 weeks. The pain sensation in the palm and the arch of the foot was measured by 4 criteria (normal, deterioration I, deterioration II, or oration 111) before the application and after the application for 6 weeks with respect to the pain sensation in the medial side of the arm using a pain/touch-pressure sensation ing device (Algesiometer; manufactured by Intercross Ltd.) in accordance with the ction manual. After the completion of the application for 6weeks, a onnaire survey on an improvement in sensation in the hands was carried out to each subject. The results are shown in Tables 10 to 13. The measurement was performed in the same manner as in Test Example 3. [003 7] TABLE 10 Hand sensation measurement (before application) Deterioration Deterioration Normal Average | H ”I vaue "n“— Hand sensation measurement (after application for 6 weeks) _-Deteripration Deterilclaration Deterifiratlon —————-7 ——————3 TABLE 11 Foot bottom sensation measurement (before application) _---- Foot bottom sensation measurement (after application for 6 weeks) ----—-Deterioration Deterioration oration Average ——-—-- -—_--- TABLE 12 Hand sensation TABLE 13 Foot bottom sensation As shown in Tables 10 to 13, the sensation in the palm and the bottom of the foot tended to be improved by applying the skin sensitivity improving cosmetic (cream) of Example product 4 . It was thus med that deterioration in sensation, particularly peripheral sensation, is expected to be improved by applying cream that includes the skin sensitivity improving agent according to the invention.

Example 5 (Preparation of skin sensitivity ing liquid nutrient composition) g of Example product 1 (milk-derived basic protein on) was dissolved in 4995 g of deionized water. The solution was stirred by a omixer (“TK ROBO MICS” manufactured by PRIMIX Corporation) at 6000 rpm for 30 minutes to obtain a basic protein fraction solution having a basic protein fraction content of 100 mg/100 g. 4.0 kg of casein, 5.0 kg of a soybean protein, 1.0 kg of fish oil, 3.0 kg of perilla oil, 18.0 kg of dextrin, 6.0 kg of a mineral mixture, 1.95 kg of a vitamin mixture, 2.0 kg of an emulsifying agent, 4.0 kg of a stabilizer, and 0.05 kg of essence were added to 5.0 kg of the milk-derived basic protein fraction solution. A retort pouch (200 ml) was filled with the mixture. The mixture was then sterilized using a retort sterilizer (class-1 pressure vessel, “RCS-4CRTGN” ctured by Hisaka Works, Ltd.) at 1216C for 20 minutes to obtain 50 kg of a skin sensitivity improving liquid nutrient composition.

The resulting skin sensitivity improving liquid nutrient ition was not observed any precipitation and the like, and did not t abnormal flavor. The skin sensitivity improving liquid nutrient composition contained the milk-derived basic protein fraction in an amount of 10 mg/100 g.

Example 6 ration of skin sensitivity improving gel-like food) 2 g ofExample product 2 (milk-derived basic protein fraction) was dissolved in 708 g of deionized water. The solution was stirred using an ultra-disperser (“ULTRA-TURRAX T-25” manufactured by IKA Japan) at 9500 rpm for 30 minutes. 40 g sorbitol, 2 g of a sour agent, 2 g of essence, 5 g of pectin, 5 g of Whey protein concentrate, l g of calcium lactate, and 235 g of deionized water were added to the solution. After ng the mixture, the mixture was filled in a cheer pack (200 ml).

After sterilizing the mixture at 85°C for 20 minutes, the pack was tightly , and thus five bags (Net 200 g each) of skin sensitivity improving gel-like food of the invention were prepared. The resulting skin sensitivity improving gel-like food was not observed any precipitation, and did not exhibit abnormal flavor. The skin ivity improving ke food contained the erived basic protein fraction in an amount of 200 mg/100 g.

Example 7 (Preparation of skin sensitivity improving drink) 2 g of sour agent was dissolved in 706 g of deionized water, and 4 g of Example product 3 (degraded milk-derived basic protein fraction) was dissolved in the solution.

The solution was d using an ultra-disperser (“ULTRA-TURRAX T-25” manufactured by IKA Japan) at 9500 rpm for 30 minutes. After the addition of 100 g ofmaltitol, 20 g ced starch syrup, 2 g of essence, and 166 g of deionized water, the resulting e was filled into a glass bottle (100 ml). After sterilizing the e at 95°C for 15 seconds, the bottle was sealed to obtain ten bottles of skin sensitivity improving drink (100 ml/ bottle). The resulting skin sensitivity improving drink was not observed any precipitation, and did not exhibit abnormal flavor. The skin sensitivity improving drink contained the degraded milk-derived basic protein on in an amount of 400 mg/100 g.

Example 8 (Preparation of skin sensitivity improving feed) 2 kg of Example product 3 (degraded milk-derived basic n fraction) was dissolved in 98 kg of deionized water. The solution was stirred using a TK-homomixer (“MARK II 160” manufactured by PRIMIX Corporation) at 3600 rpm for 40 minutes to obtain a degraded milk-derived basic protein fraction solution containing the degraded milk-derived basic protein fraction in an amount of 2 g/100 g. 12 kg of soybean cake, 14 kg of skim milk powder, 4 kg of soybean oil, 2 kg of corn oil, 23.2 kg of palm oil, 14 kg of corn starch, 9 kg of flour, 2 kg of bran, 5 kg of a vitamin mixture, 2.8 kg of ose, and 2 kg of a mineral mixture were added to 10 kg of the degraded milk-derived basic protein fraction on. The mixture was sterilized at 120°C for 4 s to obtain 100 kg of skin sensitivity improving dog food. The skin sensitivity improving dog food contained the degraded erived basic protein fraction in an amount of 200 mg/100 g.

Example 9 (Preparation of skin sensitivity ing agent (tablet)) The raw materials were mixed in the ratio shown in Table 14. l g of the resulting mixture was forrned and tableted by a conventional method to obtain a skin sensitivity improving agent of the invention. The skin sensitivity improving agent contained the milk-derived basic protein fraction in an amount of 100 mg/g.

TABLE 14 ' Hydrated crystalline glucose 83.5(wt%) erived basic protein fraction (Example 10 0 t1 ' nwwwlllllllllnlll lllllllfllll Example 10 (Preparation of skin sensitivity improving cosmetic (lotion)) A skin sensitivity improving cosmetic (lotion) was prepared by mixing the raw materials in the ratio shown in Table 15.

TABLE 15 mm Illfllll Sodium DL—pyrrolldone carboxylate mmwmm IIIEIII Parabene 0.1 Milk-derived basic protein fraction (Example t 2) Essence Proper quantity Sterilized ion-exchanged water Balance (total: 100) Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The nce in this specification to any prior publication (or ation derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this ication relates.

Claims (15)

The claims defining the invention are as follows:

1. Use of a milk-derived basic protein fraction in the cture of a ment for improving orated peripheral ion due to ageing and/or a disease.

2. The use according to claim 1, wherein the milk-derived basic protein fraction comprises lactoperoxidase and/or lactoferrin.

3. The use according to claim 1 or claim 2, n the milk-derived basic protein fraction has at least 15 wt% basic amino acids based on total amino acids.

4. The use according to any one of claims 1 to 3, wherein the milk-derived basic protein fraction is ed by bringing milk or a milk-derived raw material into contact with a cation-exchange resin to allow basic proteins to be adhered on the cation-exchange resin, and eluting a fraction adhered on the cation-exchange resin using an eluant having a sodium chloride concentration of 0.1 to 1.0 M.

5. Use of a degraded milk-derived basic protein fraction in the manufacture of a medicament for improving deteriorated peripheral sensation due to ageing and/or a disease, wherein the degraded milk-derived basic protein fraction is obtained by treating a milk-derived basic protein fraction with a protease.

6. The use according to claim 5, wherein the milk-derived basic protein fraction comprises lactoperoxidase and/or lactoferrin.

7. The use according to claim 5 or claim 6, wherein the milk-derived basic protein on has at least 15 wt% basic amino acids based on total amino acids.

8. The use ing to any one of claims 5 to 7, wherein the erived basic protein fraction is obtained by bringing milk or a milk-derived raw material into contact with a cation-exchange resin to allow basic proteins to be adhered on the -exchange resin, and eluting a fraction adhered on the cation-exchange resin using an eluant having a sodium chloride concentration of 0.1 to 1.0 M.

9. The use according to any one of claims 5 to 8, n the se is at least one protease selected from the group consisting of , trypsin, chymotrypsin, and atin.

10. Use of a milk-derived basic protein fraction in the preparation of a food, drink, feed, or cosmetic for improving deteriorated peripheral sensation due to ageing and/or a disease.

11. Use of a degraded milk-derived basic protein fraction in the preparation of a food, drink, feed, or cosmetic for improving deteriorated peripheral ion due to ageing and/or a disease, wherein the ed milk-derived basic protein fraction is obtained by treating a milk-derived basic protein fraction with a protease.

12. The use according to claim 10 or claim 11, wherein the milk-derived basic protein fraction comprises lactoperoxidase and/or lactoferrin.

13. The use according to any one of claims 10 to 12, wherein the milk-derived basic protein fraction has at least 15 wt% basic amino acids based on total amino acids.

14. The use according to any one of claims 10 to 13, wherein the milk-derived basic n fraction is obtained by bringing milk or a milk-derived raw material into contact with a cation-exchange resin to allow basic proteins to be adhered on the cation-exchange resin, and eluting a fraction adhered on the cation-exchange resin using an eluant having a sodium de concentration of 0.1 to 1.0 M.

15. The use according to any one of claims 11 to 14, wherein the protease is at least one protease selected from the group consisting of pepsin, trypsin, chymotrypsin, and pancreatin.