BR112019008683B1 - MARGARINE COMPRISING ALLULOSE - Google Patents

Abstract

The present application concerns a composition containing allulose to promote excretion of vegetable lipids from the body and a food comprising the composition of the present application and vegetable lipids.

Description

TECHNICAL FIELD CROSS REFERENCES TO RELATED ORDERS

[0001] This application claims the priority of Korean Patent Application No. 10-2016-0179487, filed on December 26, 2016, and Korean Patent Application No. 10-2016-0179488, filed on December 26, 2016, at the Korea Intellectual Property Office, the disclosures of which are incorporated herein in their entirety by reference.

TECHNICAL FIELD

[0002] The present application concerns a composition containing allulose to promote excretion of plant lipids from the body.

FUNDAMENTALS OF THE TECHNIQUE

[0003] The recommended energy intake from lipids is 20-25% in Japan and 30% in the USA and Europe (edible fats and oils: their uses and edible fats and oils, Naeha Corp., translated by Bonghyun, KIM). However, lipid consumption is increasing worldwide, and it has been reported that when high lipid consumption is continued, the risks of obesity and several adult diseases and chronic diseases may increase.

[0004] Allulose (D-psicose), which is the C-3 epimer of D-fructose, is a natural sugar present in trace amounts in commercial mixtures of D-glucose and D-fructose obtained from the hydrolysis of sucrose or isomerization of D- glucose. It has been determined to be Generally Recognized as Safe (GRAS) by the United States Department of Agriculture (USDA). Because allulose is not metabolized in the human body to be low in calories, but has 70% of the sweetness of sucrose and is therefore a sweetener that can replace sucrose, it is currently in active development. One study reported that allulose affects lipid metabolism (Yasuo nagata et al., J. Agric, Food Chem. 2015, 63, 3168-3176), but an effect of excreting lipids from the body has not been reported.

[0005] Under these circumstances, the present inventors found that allulose has an excellent fecal excretion effect of plant lipids, thus completing the present patent application.

DISCLOSURE OF THE INVENTION TECHNICAL PROBLEM

[0006] One aspect of the present application provides a composition containing allulose to promote the excretion of plant lipids from the body.

[0007] Another aspect of the present application provides a food comprising the composition of the present application and vegetable lipids.

TECHNICAL SOLUTION

[0008] Below, the present application is described in more detail. Content not described in this specification can be sufficiently recognized and inferred by those skilled in the art or similar fields of the present application, and therefore, its description will be omitted.

[0009] The respective explanations and embodiments disclosed in the present application can also be applied to other explanations and embodiments, respectively. That is, all combinations of various elements disclosed in this application fall within the scope of the present application. Furthermore, it cannot be said that the scope of the present application is limited by the specific description to be described below.

[00010] Additionally, those skilled in the art will recognize and confirm many equivalents for specific aspects of the present application described in this application using only routine experimentation. Such equivalents are also intended to be included in this application.

[00011] According to one aspect of the present invention, there is provided a composition containing allulose to promote the excretion of plant lipids from the body.

[00012] The allulose used in the present application can be, but not limited to, directly extracted from natural products or produced by chemical synthesis or biological methods. In one embodiment, the allulose contained in a cream of the present application may be liquid or crystalline allulose. Specifically, the allulose can be crystalline allulose and more specifically, the crystalline allulose can have a purity of 90% to 99.5%.

[00013] The vegetable lipids of the present application may be one or more selected from the group consisting of soybean oil, olive oil, palm oil, corn oil, palm olein oil, palm stearin oil, coconut oil, canola and sunflower oil and hydrogenated derived oils.

[00014] Furthermore, in the present application, the excretion may be triglycerides, cholesterol, free fatty acids or a combination of these. Specifically, the excretion of the present application may be fecal excretion.

[00015] The composition for promoting the excretion of plant lipids from the body of the present application may be a pharmaceutical composition, a functional food composition or a food composition.

[00016] The composition of the present application can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the desired method, and specifically, it can be administered orally.

[00017] When the composition of the present patent application is used as a pharmaceutical composition, it may further comprise one or more pharmaceutically acceptable carriers for administration, in addition to allulose. As for the pharmaceutically acceptable carrier, saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol and ethanol and a mixture of one or more components selected from these may be used and in accordance with circumstances, other common additives such as antioxidants, buffer solutions, bacteriostatic agents, etc. can be added. Diluents, dispersants, surfactants, binders and lubricants can also be additionally added to the composition of the present application, and the composition can be prepared in injectable formulations, such as aqueous solutions, suspensions or emulsions, pills, capsules, granules or tablets. The composition of the present application may be prepared in various formulations depending on the type of disease or the type of components according to any method suitable in the art or one of the methods described in Remington's Pharmaceutical Science (newest edition), Mack Publishing Company Easton PA .

[00018] The dose ranges of the pharmaceutical composition of the present application vary depending on body weight, age, sex, health condition of the patient, diet, time and mode of administration, excretion rate, severity of the disease, etc. A daily dose of allulose of the present application may be from about 0.0001 to about 600 mg/kg, specifically about 0.001 to about 500 mg/kg, and may be administered in a single dose or in multiple divided doses per day. .

[00019] The pharmaceutical composition of the present application can be used alone or in combination with surgery, hormonal therapy, drug therapy and therapies using biological response modifiers.

[00020] The composition of the present application can be used as a food or a functional food composition. When the composition of the present patent application is used as a food or functional food composition, allulose may be added as such or in combination with other foods or food ingredients and may be suitably used in accordance with any common ways. The mixing amount of the active ingredient can be determined appropriately according to the purpose of use (prevention, health or therapeutic treatment). The functional food or food composition can be, without limitation, any functional food or food composition, as long as it contains vegetable lipids. Examples of the food or functional food composition of the present application include meats, sausages, breads, cakes, chocolates, sweets, snacks, confectionery (cookies, crackers, etc.), pizza, noodles (ramen, etc.), gums, dairy products , including ice cream, various soups, ketchups, sauces, sauces, sauces, drinks, teas, beverages, alcoholic beverages, vitamin complexes, etc.

[00021] In addition to the above ingredients, the food or functional composition of the foods of the present application may also contain a variety of nutritional supplements, vitamins, electrolytes, flavorings, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents for carbonated drinks, etc. These ingredients can be used independently or in combination. The proportion of the additives can be selected from the range of 0.01 to 0.20 parts by weight, relative to 100 parts by weight of the food or functional composition of the present application.

[00022] According to another aspect of the present application, a food comprising the composition of the present application and vegetable lipids is provided. Furthermore, in the food of the present application, allulose may be contained in an amount of 20 to 150 parts by weight based on 100 parts by weight of vegetable lipids.

[00023] Specifically, the foods of the present application can be margarine or cream. More specifically, the margarine of the present application may further include one or more ingredients selected from the group consisting of water, sodium chloride, milk, lecithin, an organic acid and an emulsifier. Furthermore, the substitute of the present application may also include casein, maltose and phosphate. Furthermore, the cream of the present application may be in powder form, and specifically, may have a water content of 0.5% to 5%. In one embodiment, the cream of the present application may be coffee cream or tea cream.

[00024] In the foods of the present application, allulose can be contained in an amount of 20 to 150 parts by weight, based on 100 parts by weight of vegetable lipids. Specifically, the allulose content may be contained in an amount of 20 to 100 parts by weight, 20 to 50 parts by weight, 20 to 40 parts by weight, 30 to 150 parts by weight, 30 to 100 parts by weight, 30 to 50 parts by weight, or 30 to 40 parts by weight, based on 100 parts by weight of plant lipids.

[00025] In one embodiment, the food of the present application cannot contain sugar.

[00026] In the food of the present application, the descriptions of the composition, vegetable lipids and foods are as described in the aspects mentioned above.

[00027] However, according to another aspect of the present application, there is provided a method for promoting the excretion of plant lipids administered to a patient from the body, the method comprising a step of administering the plant lipids to the individual; and a step of administering allulose to the patient before, after or simultaneously with the step of administering the plant lipids to the patient. The patient can be a human or an animal.

[00028] In the method of the present application, 20 to 150 parts by weight of allulose can be administered based on 100 parts by weight of plant lipids consumed by the individual. Specifically, 20 to 100 parts by weight, 20 to 50 parts by weight, 20 to 40 parts by weight, 30 to 150 parts by weight, 30 to 100 parts by weight, 30 to 50 parts by weight, or 30 to 40 parts by weight Allulose weight can be administered based on 100 parts by weight of the advantageous lipids. Administration can be oral administration.

[00029] In the method of the present application, the descriptions of allulose, plant lipids, administration and excretion are as described in the above-mentioned aspects.

[00030] According to another aspect of the present application, there is provided a use of an allulose-containing composition to promote the excretion of vegetable lipids contained in a food.

[00031] In the use of the present application, the descriptions of allulose, the food, the vegetable lipids and the excretion are as described in the aspects mentioned above.

ADVANTAGEOUS EFFECTS

[00032] In the present application, it was found that allulose supplied together with vegetable lipids notably increased fecal lipid excretion. Consequently, the present application has the effect of reducing consumer concerns regarding high lipid consumption due to the ingestion of plant lipids or foods containing them.

BRIEF DESCRIPTION OF THE DRAWINGS

[00033] Figure 1 shows the figure-eight body changes observed in C57BL/6J mice fed a high-fat diet (HFD) containing plant lipids along with allulose for 8 weeks. In Figs. 1a to 1d, abbreviations for a control group represent the following: OL: olive oil supply, MG: margarine supply, PO: palm oil supply, SO: soybean oil supply. In Figs. 1a to 1d, the abbreviations for a test group are as follows: OLA: olive oil + 5% store supply, p/w, MGA: margarine + 5% store supply, p/w, POA: palm oil + 5% allulose supply w/w, SOA: soybean oil + 5% allulose supply, w/w.

[00034] Meanwhile, Fig. 1e shows changes in body weight observed in C57BL/6J mice fed a high-fat diet (HFD) containing cream over allulose for 8 weeks. Here, RP for a control group represents provision with a high-fat diet (HFD) together with cream and PRA represents provision with a high-fat diet (HFD) + cream + 5% allulose (w/w).

[00035] Fig. 2a shows changes in fecal lipid excretion after 8 weeks observed in C57BL/6J mice fed a high-fat diet (HFD) containing plant lipids along with allulose. Here, data were represented as means ± SE. The significance between the no allulose group and the 5% allulose group is as follows: * p <0.05, ** p <0.01, *** p <0.001. In Fig. 2a, each abbreviation represents the following: OLA, HFD + olive oil + 5% allulose (w/w); OL, HFD + olive oil; MGA, HFD + margarine + 5% allulose (w/w); MG, HFD + margarine; POA, HFD + palm oil + 5% allulose (w/w); PO, HFD + palm oil; SOA, HFD + soybean oil + 5% allulose (w/w); OL, HFD + soybean oil.

[00036] Fig. 2b shows changes in fecal lipid excretion after 8 weeks observed in C57BL/6J mice fed a high-fat diet (HFD) containing cream together with allulose. Here, data were represented as means ± SE. The significance between the group without allulose and the group with 5% allulose is as follows: * p <0.05, ** p <0.01, *** p <0.001. In fig. 2, PRA represents "HFD + cream + 5% allulose (w/w)" and PR represents "cream + HFD".

MODE OF CARRYING OUT THE INVENTION

[00037] Below, preferred examples are presented to help understand the present application. However, the following examples are provided to facilitate the understanding of the present application, and the content of the present application is not limited by examples.

Experimental methods 1. Experimental animal breeding 1-1. Palm oil, olive oil, soybean oil and margarine

[00038] Twenty C57BL/6J mice (males, 4 weeks old) were purchased from The Jackson Laboratory (USA) and used. After the animals were acclimated to the feeding environment with a "lab-chow" diet (Purina co., USA) for 4 weeks, they were divided in a randomized block design into two groups: a negative control without allulose intake ( n = 10/one type of lipid); and a test group with allulose intake (n = 10/one type of lipid) and were fed the diets for 8 weeks.

[00039] For the negative control group, the high-fat diet was applied to the AIN-76 diet, and vegetable lipids [palm oil (Wilmar, Malaysia), olive oil (CJ CheilJedang Corp., South Korea), soybean (CJ C heilJedang Corp., South Korea), margarine (Ottogi Foods Co., Ltd., 'Corn margarine', raw materials: consisted of 80% by weight vegetable lipids (palm olein oil, of palm stearin, coconut oil, corn oil)] For the test group, among the dietary ingredients for the negative control group, 5% by weight of sucrose was replaced by allulose (crystalline allulose, 98% by weight of allulose based on dry solids, CJ CheilJedang Corporation) (Table 1) All animal experiments were carried out under the approval of the Ethics Committee for Animal Studies of Kyungpook National University, South Korea (approval no. KNU-2013- 18). [Table 1] Composition of the test feed (% of diet, w/w) [Table 1]

[00040] Note 1) Mineral mixture: AIN-76 mineral mixture (gram/kg): calcium phosphate, 500; sodium chloride, 74; potassium citrate, 2220; potassium sulfate, 52; magnesium oxide, 24; manganous carbonate, 3.5; ferric citrate, 6; zinc carbonate, 1.6; cupric carbonate, 0.3; potassium iodate, 0.01; sodium celenite, 0.01; chromium potassium sulfate, 0.55; sucrose 118.03.

[00041] Note 2) Vitamin mixture: AIN-76 vitamin mixture (gram/kg): thiamine HCL, 0.6; riboflavin, 0.6; Pyridoxine HCl, 0.7; nicotic acid, 0.003; D-calcium pantothenate, 0.0016; folate, 0.2; D- biotin, 0.02; cyanocobalamin (vitamin B12), 0.001; retinyl palmitate premix, 0.8; DL-alpha-tocopheryl acetate, premix, 20; cholecalciferol (vitamin D3), 0.0025; menaquinone (vitamin K), 0.05; antioxidant, 0.01; fine powder sucrose, 972.8

1-2. Cream

[00042] Sixteen C57BL/6J mice (males, 4 weeks old) were purchased from The Jackson Laboratory (USA) and used. After the animals were acclimatized to the feeding environment with a “lab-chow” diet (Purina co., USA) for 4 weeks, they were divided in a randomized block design into two groups: negative control group without allulose intake (RP, n = 8); and a test group with allulose intake (PRA, n = 8) and were fed the diets for 8 weeks.

[00043] For the negative control group, the high-fat diet was applied to the AIN-76 diet, and as the vegetable lipid, cream [Dongsuh foods corporation 'Prima', South Korea, raw materials: hydrogenated vegetable oil 30 -38% by weight (hydrogenated coconut oil, hydrogenated palm oil), starch syrup (including maltose), disodium casein, potassium phosphate, dibasic, calcium phosphate, tribasic]. For the test group, among the dietary ingredients for the negative control group, 5 wt% sucrose was replaced by allulose (crystalline allulose, 98 wt% aluminous with dry solids, CJ CheilJedang Corporation) (Table 2). All animal experiments were performed under the approval of the Ethics Committee for Animal Studies of Kyungpook National University, South Korea (approval no. KNU-2013-18). [Table 2] Composition of the test feed (% of diet, w/w) [Table 2] Note 1) Mineral mixture: is the same as in Table 1 above and will therefore be omitted here. Note 2) Vitamin Blend: This is the same as in Table 1 above and will therefore be omitted here.

[00044] All groups were allowed to ingest iso-energetic diet of the same level through pair feeding based on the test group, so as to exclude the calorie reducing effect due to allulose and the diets were kept refrigerated at 4 °C during the feeding period. The animals were housed in an individual cage under constant temperature (25 ± 2 °C), constant humidity (50 ± 5%) and a 12-hour light-dark cycle.

2. Measurement of intake and body weight

[00045] Dietary intake was measured at a constant time each day, and body weight was measured at a constant time each week.

3. Collection and analysis of fecal samples 3-1. Fecal sample collection

[00046] Feces were collected for 84 hours (3.5 days) after the end of feeding, dried and stored in the freezer.

3-2. Extraction of fecal lipids

[00047] For measurement, neutral lipids, cholesterol and free fatty acids in feces were extracted, modifying and complementing the method of Folch et al (1957). Specifically, the dried feces were ground in a mortar to obtain 0.5 g and then 5 mL of a chloroform:methanol solution (2:1, v/v) was added and the lipids were extracted at 4 °C for 24 hours. The extract was centrifuged at 3000 x g (4 °C) for 10 minutes, and then 3 ml of the supernatant was removed and dried under nitrogen gas at 37 °C and then dissolved again in 1 ml of the same extraction solvent. 200 μL of each sample for measuring neutral lipids, cholesterol and free fatty acids was removed, dried again under nitrogen gas and the samples for measuring neutral lipid and total cholesterol were dissolved in 500 μL of ethanol. The sample for measuring free fatty acids was dissolved in 2.25 ml of NaOH and then a 1M HCl solution was added to adjust the pH to the range of 2-3. To remove haze occurring from development with 3 mM cholic acid (sodium salt), 0.5% Triton X-100 as an emulsifier was mixed with enzyme test solution during quantification of total cholesterol and neutral lipids. .

3-3. Quantification of total cholesterol in feces

[00048] To measure total cholesterol, 10 μL of a solution dissolved in 500 μL of ethanol and 690 μL of the emulsifier were mixed and 800 μL of a test solution for measurement (kit, Asan Pharmaceutical Co., Ltd.) for which the enzymatic method of Allain et al. (1974) was applied mixed with the same. For the quantification of free cholesterol (FC) and ester cholesterol (EC) in feces, EC was converted by cholesterol esterase into FC and fatty acids. Among them, FC was subjected to reaction with cholesterol oxidase to convert into Δ4-cholestenone. The resulting product and H2O2 which is the substrate were subjected to reaction with peroxidase, phenol and 4-amino-antipyrine to obtain a color revealing material and the absorbance was measured at 500 nm. Quantification was calculated by comparing the measured value with the cholesterol standard curve.

3-4. Quantification of fecal neutral lipids

[00049] For the measurement of neutral lipid, 10 μL of a solution dissolved in 500 μL of ethanol and 690 μL of the emulsifier were mixed and 800 μL of a test solution for measurement (kit, Asan Pharmaceutical Co., Ltd.) the enzymatic method of McGowan et al. (1983) was applied was mixed with the same. The neutral lipid was degraded by lipoprotein lipase into glycerol and fatty acids. From the degradation products, glycerol was formed, through ATP and the action of glycerol kinase (GK), L-α-glycerol phosphate, which reacted with O2 and glycerophosphoxidase (GPO) to produce H2O2. Peroxidase and 4-amino-antipyrene were treated to develop a red color and then the absorbance was measured at 550 nm and quantification was performed by comparison with the glycerol standard curve.

3-5. Quantification of fecal free fatty acids

[00050] The concentration of free fatty acids was measured using a test solution for measuring free fatty acids (n-esterified fatty acid; NEFA kit, Wako, Japan) according to the principle of color development using an enzyme mode . First, acyl-coenzyme A synthase was applied to free fatty acid in plasma to produce acyl-CoA, AMP, and pyrophosphoric acid. Then, acyl-coenzyme A oxidase was added to generate 2,3-trans-enoyl-CoA and hydrogen peroxide. Peroxidase, 4-aminoantipyrine, and N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine were treated with it to develop the red color, and then the absorbance was measured at 546 nm and the Quantification was performed by comparison with the free fatty acid standard curve.

Experimental results 1. Confirmation of the effect of inhibiting body weight gain due to allulose 1-1. Palm oil, olive oil, soybean oil and margarine

[00051] At the time of week 0 of the diet, the body weights of the negative control group and the test group were similar (Table 3), but after 8 weeks of the diet, the effect of inhibiting body weight gain in the control group test was confirmed in comparison to the negative control group (Table 4 and Figs. 1a to 1d). Specifically, in the case of the MGA group and the OLA group, from the 5th week of the diet and, in the case of the POA group, from the 6th week of the diet, the significant effect of inhibiting weight gain was confirmed in comparison with the negative control group (Figs. 1a to 1c). Particularly, in the case of the SOA group, the effect of inhibiting body weight gain was exhibited from the 1st week of the diet, and therefore, the exceptional effect of inhibiting weight gain was confirmed in comparison with other plant lipids (Fig. 1d). [Table 3]

[00052] Data were represented as means ± SE. *ANOVA is a comparative analysis between four groups. ** The t-test is the comparison value between the HF D without allulose and the HFD containing 5% by weight of allulose in each group. [Table 4]

[00053] Data were represented as means ± SE. *ANOVA is a comparative analysis between four groups. **The t-test is the comparison value between the HFD without allulose and the HFD containing 5% by weight of allulose in each group (# p <0.05, ### p <0.001).

1-2. Cream

[00054] At week 0 of the diet, the body weights of the negative control group (RP) and the test group (PRA) were similar (Table 5). However, the body weight of the negative control group increased significantly from 1 week onwards, while the body weight of the test group was notably inhibited from the 1st week of the diet and thus after 8 weeks of the diet, the significant effect of inhibiting body weight gain in the test was confirmed (Table 5 and Fig. 1e). [Table 5]

[00055] Data were represented as means ± SE. ** t-test is the comparison value between PR without allulose and PRA containing 5% by weight of allulose in each group.

2. Confirmation of the effect of excreting plant lipids due to allulose 2-1. Palm oil, olive oil, soybean oil and margarine

[00056] The effect of excreting plant lipids due to allulose was confirmed by lipid excretion in feces.

[00057] As a result, it was confirmed that the content of triglycerides and free fatty acids in feces were significantly increased in the test groups of all types of oils, compared to the negative control group. Particularly, it was confirmed that the content of free fatty acids in the test groups was significantly higher than that in the negative control group (Tables 6 to 9 and Figs. 2a (A) and (C)). Furthermore, it was confirmed that the SOA group also showed a significant increase in fecal cholesterol content compared to the SO group (Fig. 2a(B)). [Table 6]

[00058] Data represented as means ± SE. There was a significant difference between OLA p <0.01. HELLO, Olive Oil + HFD + 5% olive oil. [Table 7]

[00059] means ± SE. There was *** p <0.001. MGA, + margarine. The data were significant difference HFD + margarine + 5% [Table 8]

[00060] Data were represented as means ± SE. There was a significant difference between POA and PO: ** p <0.01, *** p <0.001. POA, HFD + palm oil + 5% allulose; PO, HFD + palm oil. [Table 9]

[00061] Data were represented as means ± SE. There was a significant difference between SOA and SO: * p <0.05. SOA, HFD + soybean oil + 5% allulose; SO, HFD + soybean oil.

2-2. Cream

[00062] The effect of lipid excretion in the cream due to allulose was confirmed by lipid excretion in feces.

[00063] As a result, it was confirmed that the content of triglycerides and free fatty acids in feces were significantly increased in the test group compared to the negative control group. Particularly, it was confirmed that the content of free fatty acids in the test group was significantly higher than that in the negative control group (Tables 10 and Fig. 2b). [Table 10]

[00064] Data were represented as means ± SE. There was a significant difference between PR and PRA: * p <0.05, *** p <0.001. PRA, HFD + cream + 5% allulose; PR, HFD + cream.

2-3. Consideration

[00065] It was confirmed that when allulose was consumed together with vegetable lipids in cream, fecal excretion of lipids was promoted. Furthermore, it was confirmed that when allulose was consumed together with palm oil, olive oil, soybean oil and margarine, fecal lipid excretion was also promoted. Especially, when allulose was taken together with soybean oil, the effect of lipid excretion was excellent compared with taking allulose together with other plant lipids and the excretion of cholesterol was also increased.

[00066] The above-mentioned description of the present application is intended to be merely exemplary, and it will be understood by those skilled in the art to which the present application belongs that the present application can be easily modified to other specific forms without changing the technical concepts. or its essential characteristics. It is, therefore, to be understood that the above-described embodiments are illustrative in all respects and not restrictive.

Claims (1)

1. Margarine characterized by the fact that it comprises allulose, vegetable lipids and one or more of water, sodium chloride, milk, lecithin, an organic acid, or an emulsifier; wherein the vegetable lipids are one or more of soybean oil, olive oil, palm oil, palm olein oil, palm stearin oil, coconut oil, and hydrogenated versions of these oils; and wherein allulose is contained in an amount of 20 to 150 parts by weight based on 100 parts by weight of plant lipids, based on dry solids.