Inhibiting reduction of lean body mass and inhibiting accumulation of liver fat by administering allulose

Abstract

The present invention relates to a composition for inhibiting liver fat accumulation comprising allulose as an active ingredient and a pharmaceutical composition and a food composition using thereof.

Claims

1. A method for inhibiting reduction of lean body mass and inhibiting accumulation of liver fat comprising administering, to an overweight subject having a BMI of 23 kg/m.sup.2 or more to less than 30 kg/m.sup.2, a daily dose of allulose at 0.05 to 0.75 g per kg of the subject, wherein the average change of the lean body mass measured before and after administration is 1.5% or less at 12 weeks.

2. The method according to claim 1, wherein the daily dose of the allulose is administered at an amount of 3 to 45 g to the subject.

3. The method according to claim 2, wherein the daily dose of the allulose is administered at an amount of 3 to 25g to the subject.

4. The method according to claim 2, wherein the daily dose of the allulose is administered at an amount of 6 to 12g to the subject.

5. The method of claim 1, wherein the daily dose of the allulose is administered by splitting the dose 1 to 4 times.

6. The composition according to claim 1, wherein the administration is oral administration.

7. The composition according to claim 1, wherein the allulose is a component in a mixed saccharide syrup, or in a powder obtained from a mixed saccharide syrup, that comprises 2 to 55 parts by weight of allulose, 30 to 80 parts by weight of fructose, 2 to 60 parts by weight of glucose and 0 to 15 parts by weight of an oligosaccharide, based on the total solid content of the mixed saccharides.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a schematic diagram showing the human application method according to An embodiment of the present invention.

MODE FOR INVENTION

(2) Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are intended to illustrate the present invention only, but the scope of the present invention is not limited by these examples.

Preparative Example 1. Preparation of Allulose

(3) The allulose powder used in the following example was prepared by the method described in Korean Patent Publication No. 2014-0021974, Korean Patent Publication No. 2014-0080282 and Korean Granted Patent No. 10-1318422.

Example 1: Efficacy Test

(4) (1) Subject Selection

(5) After conducting a screening test for those who heard the detailed description of the purpose and research method of the present example and agreed in writing as subjects, those who met the selection/exclusion criteria were selected. The specific selection criteria are as follows, and the final selected subjects are 99 men and women who are overweight or obese, and the demographic information of the selected subjects is shown in the following Table 1.

(6) Selection Criteria

(7) 1. Adult men and women between ages of full 19 to 75

(8) 2. Overweight adult men and women with BMI of 23 kg/m2 or more to less than 30 kg/m2

(9) 3. In case of women of childbearing age, negative in pregnancy test during screening

(10) 4. Patients that can be controlled (diabetes or hypertension, etc.): patients that are well controlled with drugs are included, and 10% of the subjects have hypertension and 5% have diabetes.

(11) The gender of the subject participating in the present human application test were 5 men (15.15%) and 28 women (84.85%) in the high-dose group, 6 men (18.18%) and 27 women (81.82%) in the low-dose group, and 2 men (6.06%) and 31 women (93.94%) in the control group. The average age was 45.06 years old (range: 25-65 years old) in the high-dose group, 41.52 years old (range: 26-61 years old), and 44.61 years old (range: 26-63 years old) in the control group, and both gender and age did not show a statistically significant difference in the subject distribution.

(12) By dividing 99 selected subjects into 3 groups, the test subjects were finally determined as the high-dose group (33 people), low-dose group (33 people) and control group (33 people).

(13) TABLE-US-00001 TABLE 1 High-dose Low-dose group group Control p-value ITT 33 33 33 Age Mean 45.06 41.52 44.61 0.3698 SD 12.06 10.31 11.00 Median 42.00 40.00 46.00 Min 25.00 26.00 26.00 Max 65.00 61.00 63.00 Gender Male  5 (15.15)  6(18.18) 2 (6.06)  0.4184.sup.1) Female 28 (84.85) 27(81.82) 31(93.94) p-value: Kruscal-wallis test .sup.1)Fisher's exact test

(14) (2) Test Products

(15) The efficacy test was performed by dividing 99 selected test subjects into 3 groups, and by providing three products to the high-dose group (33 people), low-dose group (33 people), and control group (33 people), respectively. The allulose high-dose product was provided to the high-dose test group, and the allulose low-dose product was provided to the low-dose test group, and placebo product was provided to the control group.

(16) The products were stored in airtight containers, at a room temperature (1-30° C.), in a dry and well-ventilated place.

(17) TABLE-US-00002 TABLE 2 Product allulose allulose name high-dose low-dose placebo component allulose allulose, anhydrous name anhydrous glucose glucose type and white white white formulation crystalline crystalline crystalline powder powder powder amount of raw allulose allulose anhydrous material 12 g/d 6 g/d, glucose product anhydrous 12 g/d glucose 6 g/d packaging unit 6 g/bag 6 g/bag 6 g/bag

(18) (3) Intake Method

(19) Ninety nine (99) subjects were randomly assigned to the high-dose group, low-dose group or control group, with including 33 subjects in each test group. In three test groups, the products were mixed in 100 ml of water and were administered orally twice a day.

(20) As the daily dose for one test subject, 12 g allulose in the high-dose group, 6 g allulose and 6 g anhydrous glucose in the low-dose group, and 12 g anhydrous glucose in the control group were divided and provided twice a day. It was administered from the day after the random assignment day (0 day), and was taken with drinking water twice at morning and evening with meals for 12 weeks.

(21) At the random assignment day (0 day), the efficacy evaluation items and safety evaluation items such as DXA measurement and the like were checked, and the evaluation items were also checked through hospital outpatient visit at 4 weeks, 8 weeks and 12 weeks from the day of beginning administration of test products. In addition, during the human application test, the usual physical activity of subjects was investigated using Korean version of International Physical Activity Questionnaire (IPAQ). During the human application test period, the subjects were trained to maintain the activity amount being similar to normal physical activity. At each hospital outpatient visit, the amount of meal intake was examined by the recording board made by the subjects. The subjects were trained to maintain the amount of meal intake being similar to the usual amount with avoiding overeating condition.

Example 2: Efficacy Evaluation Method

(22) (1) Body Fat and Lean Body Mass Measured by DXA

(23) At the time of second hospital visit (visit 2) and final hospital visit (visit 5), the body fat ratio and body fat were measured using prodigy DXA of U.S. Lunar Company.

(24) For the body fat measured by DXA, after calculating the change of the body fat after 12 weeks compared to the base time, technical statistics (mean, standard deviation, median value, minimum value, maximum value) were obtained. The comparative statistic test for the three groups was performed using ANOVA or Kruscal-Wallis test by testing whether data were normally distributed. When there was a significant difference between the three groups, the multiple comparison was conducted through Tukey-Kramer method. In order to find out whether there was a difference in the mean change in a group, it was analyzed using Paired t-test or Wilcoxon's signed rank test.

(25) (2) Glucose Index Test

(26) At the time of screening hospital visit (visit1), visit2 and final visit (visit5), FBS, Insulin, and HbAlc test was conducted for the collected blood.

(27) After calculating the average change of the glucose index (FBS, Insulin, HbAlc) after 12 weeks compared to the base time, technical statistics (mean, standard deviation, median value, minimum value, maximum value) were obtained. The comparative statistic test for the three groups was performed using ANOVA or Kruscal-Wallis test by testing whether data were normally distributed. When there was a significant difference between the three groups, the multiple comparisons were conducted through Tukey-Kramer method. In order to find out whether there was a difference in the mean change in a group, it was analyzed using Paired t-test or Wilcoxon's signed rank test.

(28) (3) Liver CT

(29) In order to evaluate the degree of fat deposition in the liver at the time of hospital visit 2 and visit 5, the HU of liver, HU of spleen, HU difference of liver and spleen (Liver minus Spleen), and HU ratio of liver and spleen (Ratio Between Liver and Spleen) after 12 weeks compared to the base time were measured. CT attenuation values (Hounsfield units) were measured in regions of interest (ROI) selected as a homogeneous place avoiding blood vessels in the right lobe of liver and spleen in one non-contrast-enhanced abdominal CT image including both liver and spleen three times, and the average value was taken as each CT attenuation number. Then, the size of each region of interest was designated equally as 120 mm.sup.2.

(30) Through liver CT, for HU of liver, HU of spleen, HU difference of liver and spleen (Liver minus Spleen), and HU ratio of liver and spleen (Ratio Between Liver and Spleen) after 12 weeks, technical statistics (mean, standard deviation, median value, minimum value, maximum value) were proposed for the average change compared to the base time. The comparative statistic test for the three groups was performed using ANOVA or Kruscal-Wallis test by testing whether data were normally distributed. When there was a significant difference between the three groups, the multiple comparisons were conducted through Tukey-Kramer method. In order to find out whether there was a difference in the mean change in a group, it was analyzed using Paired t-test or Wilcoxon's signed rank test.

(31) (4) Blood chemical test result The test items performed at screening visit (visit1), visit2 and end visit (visit5) were Alkaline phosphatase, AST, ALT, and γ-GTP.

(32) In case of continuous data, for the average change compared to the base time, technical statistics (mean, standard deviation, median value, minimum value, maximum value) were proposed for the average change compared to the base time. The comparative statistic test for the three groups was performed using ANOVA or Kruscal-Wallis test by testing whether data were normally distributed. When there was a significant difference between the three groups, the multiple comparison was conducted through Tukey-Kramer method. In order to find out whether there was a difference in the mean change in a group, it was analyzed using Paired t-test or Wilcoxon's signed rank test. In case of categorical data, technical statistics (frequency, percentage) were proposed, and comparative statistic test was performed between three groups using Chi-square test or Fisher' exact test.

Example 3: Efficacy Evaluation Result

(33) (1) Body Fat and Lean Body Mass Measured by DXA

(34) For 99 8ITT analysis subjects, the average change in bod fat at 12 weeks compared to the base time was increased by 0.25±1.34 kg in the high-dose group and was increased by 0.16±0.91 kg in the low-dose group, and was increased by 0.95±1.65 kg in the control group, and it showed a statistically significant difference between three groups (p-value=0.0358). In addition, the control group showed a statistically significant difference before and after at 12 weeks compared to the base time (p-value=<0.0001).

(35) The average change of the lean item did not show a statistically significant difference between three groups, and in the control group, the average change at 12 weeks compared to the base time was reduced by −0.61±1.28 kg, and thereby showed a statistically significant difference (p-value=0.0048). The result of body fat measured by DXA was shown in the following Table 3.

(36) TABLE-US-00003 TABLE 3 High-dose Low-dose group group Control p-value.sup.(a) ITT 33 33 33 Fat (Kg) Mean 26.06 26.64 26.18 0.8735 Baseline SD 4.36 4.74 5.28 12 W Mean 26.31 26.80 27.13 0.7920 SD 4.59 4.76 5.41 Difference 0.25 ± 1.34 0.16 ± 0.91 0.95 ± 1.65 0.0358.sup.1)+ p-value.sup.(b) 0.2937 0.3177 <0.0001.sup.2) Lean(Kg) Mean 40.46 42.21 39.74 0.7691.sup.1) Baseline SD 6.94 9.12 6.64 12 W Mean 40.24 42.26 39.13 0.4607.sup.1) SD 6.75 9.40 6.63 Difference −0.22 ± 1.08  0.05 ± 0.89 −0.61 ± 1.28  0.0884.sup.1) p-value.sup.(b) 0.2522 0.7373 0.0048.sup.2) p-value.sup.(a): ANOVA p-value.sup.(b): Paired t-test .sup.1)Kruskal-wallis test .sup.2)Wilcoxon's signed rank test .sup.+P-value < 0.05, statistically significant, post hoc method by tukey low-dose group versus control group

(37) For the body fat item measured by DXA, the variance at 12 weeks compared to the base time showed a statistically significant difference between three groups. In the control group, the increase of the body fat showed a statistically significant difference before and after administration. However, the allulose low-dose group and high-dose group were slightly increased, but there was no statistically significant difference before and after administration.

(38) The lean body mass was reduced by −0.61±1.28 kg after 12 weeks in the control group, but the allulose administration groups did not show a difference before and after administration. As overweight was included in selection criteria of subjects in this study, all subjects were in mild obesity group (average body mass index is 27 or less), and it is considered that there was no statistically significant change in weight or body mass index due to the lack of subjects per group. The reduction of muscle mass was suppressed.

(39) (2) Glucose Index

(40) As the result of the glucose index test for 99 ITT analysis subjects, the difference of the average change at 12 weeks compared to the base time between the high-dose group, low-dose group and control group was not significant for all the items of Glucose, Insulin, and HbAlc, and there was no statistically significant difference between the groups before and after, so there was no hypoglycemic effect in the analysis subjects as normal blood sugar. The result of glucose index analysis of analysis subjects was shown in the following Table 4, Table 5 and Table 6.

(41) TABLE-US-00004 TABLE 4 High-dose Low-dose group group Control p-value.sup.(a) ITT 33 33 33 Glucose(mg/dl) Mean 94.70 94.76 93.85 0.8542 Baseline SD 9.72 10.16 12.21 Median 93.00 94.00 92.00 Min 76.00 78.00 80.00 Max 122.00 128.00 145.00 12 W Mean 94.58 97.18 92.64 0.4407 SD 10.62 13.72 11.93 Median 94.00 97.00 91.00 Min 78.00 79.00 66.00 Max 126.00 140.00 139.00 Difference −0.12 ± 8.35 2.42 ± 10.10 −1.21 ± 8.36 0.3770 p-value.sup.(b) 0.9341.sup.2) 0.2410 0.4110.sup.2) p-value.sup.(a): Kruskal-wallis test p-value.sup.(b): Wilcoxon's signed rank test 1) ANOVA .sup.2)Paired t-test

(42) TABLE-US-00005 TABLE 5 High-dose Low-dose group group Control p-value.sup.(a) ITT 33 33 33 Insulin (mg/dl) Mean 6.95 7.92 6.75 0.4110 Baseline SD 3.01 5.84 3.77 Median 6.20 6.50 5.30 Min 3.10 2.40 2.00 Max 18.20 36.20 20.00 12 W Mean 8.70 9.18 6.65 0.1273 SD 5.34 9.94 3.14 Median 7.50 7.10 5.80 Min 2.10 2.30 2.40 Max 31.20 61.30 13.80 Difference 1.75 ± 5.33 1.27 ± 5.15 −0.10 ± 2.60 0.4399 p-value.sup.(b) 0.0629 0.0814 0.4801 p-value.sup.(a): Kruskal-wallis test p-value.sup.(b): Wilcoxon's signed rank test 1) ANOVA 2) Paired t-test

(43) TABLE-US-00006 TABLE 6 High-dose Low-dose group group Control p-value.sup.(a) ITT 33 33 33 HbA1 (mg/dl) Mean 5.61 5.57 5.52 0.4325 Baseline SD 0.36 0.44 0.43 Median 5.60 5.50 5.40 Min 5.00 4.80 5.00 Max 6.60 7.20 6.90 12 W Mean 5.66 5.64 5.56 0.4527 SD 0.35 0.49 0.43 Median 5.60 5.60 5.50 Min 5.00 4.90 5.00 Max 6.50 7.20 6.90 Difference 0.05 ± 0.18 0.07 ± 0.22 0.04 ± 0.14 0.8846.sup.1) p-value.sup.(b) 0.1159 0.0915 0.0996.sup.2) p-value.sup.(a): Kruskal-wallis test p-value.sup.(b): Wilcoxon's signed rank test .sup.1)ANOVA .sup.2)Paired t-test

(44) (3) Liver CT

(45) As the result of the CT test tested for 99 ITT analysis subjects, the average change at 12 weeks compared to the base time did not show a statistically significant difference between three groups in all the items. All the Liver HU average value, Liver−Spleen HU, and Liver/Spleen HU items showed a decreasing tendency in the liver CT, and showed a statistically significant difference before and after (p-value=0.0300, 0.0027, 0.0029).

(46) TABLE-US-00007 TABLE 7 High-dose Low-dose group group Control p-value.sup.(a) ITT 33 33   33 Liver CT Liver HU Mean(HU) Base line Mean(HU) 58.27 55.86 59.62 0.9676 SD 10.30 14.86 7.12 12 W Mean 57.49 55.17 57.77 0.6497 SD 11.21 15.36 7.57 Difference −0.78 ± 5.60 −0.69 ± 5.17 −1.85 ± 4.68 0.3310 p-value.sup.(b) 0.4304.sup.2)   0.5903 0.0300.sup.2) Spleen HU Mean(HU) Base line N 33 32 §  33 Mean 49.80 51.07 49.86 0.2269.sup.1) SD 3.40  2.65 3.81 12 W Mean 50.30 50.66 50.76 0.8207.sup.1) SD 3.16  3.06 3.10 Difference  0.50 ± 2.77 −0.41 ± 2.34  0.90 ± 3.37 0.0676 p-value.sup.(b) 0.3069.sup.2)   0.1549 0.1354.sup.2) Liver - Spleen HU.sup.+ Base line N 33 32 §  33 Mean 8.47  5.30 9.76 07957 SD 9.72 14.86 7.87 12 W Mean 7.19  4.75 7.01 0.8278 SD 10.94 15.15 8.04 Difference −1.28 ± 5.36 −0.55 ± 5.43 −2.75 ± 4.85 0.2277 p-value.sup.(b) 0.1802.sup.2)    0.5726.sup.2) 0.0027.sup.2) Liver/Spleen (HU) .sup.++ Base line N 33 32 §  33 Mean 1.17  1.11 1.20 0.6829 SD 0.20  0.30 0.17 12 W Mean 1.14  1.09 1.14 0.8544 SD 0.23  0.31 0.16 Difference −0.03 ± 0.11 −0.01 ± 0.11 −0.06 ± 0.11 0.1204 p-value.sup.(b) 0.1903.sup.2)   0.8942 0.0029.sup.2) .sup.+Difference of Liver and Spleen (Liver minus Spleen) .sup.++ Ratio of Liver and Spleen (Ratio between Liver and Spleen) § R012 subject was excluded from the test, because the spleen scan was not measured at the base time. p-value.sup.(a): Kruskal-wallis test p-value.sup.(b): Wilcoxon's signed rank test .sup.1)ANOVA .sup.2)Paired t-test

(47) (4) Blood Chemical Test Result

(48) The difference of the average change at 12 weeks compared to the base time by each blood chemical test items between the groups was analyzed, and Alkaline phosphatase (p-value=<0.0001), ALT (SGPT) (p-value=0.0127), and γ-GTP (p-value=0.0043) in the blood chemical test items showed a statistically significant difference between three groups.

(49) The Alkaline phosphatase and γ-GTP showed a statistically significant difference in the three groups, as they were reduced in the high-dose group and low-dose group and a statistically significant difference before/after administration in the high-dose group and low-dose group, and they were increased in the control group. ALT showed a statistically significant difference between three groups, as it was reduced in the high-dose group and low-dose group and was increased in the control group, and a statistically significant difference before/after administration in the high-dose group.

(50) TABLE-US-00008 TABLE 8 High-dose Low-dose group group Control p-value ITT 33 33 33 Blood chemical test Alkaline Mean 183.21 178.97 175.03 0.9119.sup.1) phosphatase(U/L) SD 56.52 49.53 41.85 Median 169.00 170.00 165.00 Min 104.00 102.00 112.00 Max 331.00 328.00 270.00 AST(SGOT)(U/L) Mean 18.76 22.64 19.24 0.5445.sup.1) SD 5.33 10.52 6.66 Median 18.00 18.00 17.00 Min 10.00 12.00 10.00 Max 28.00 53.00 41.00 ALT(SGPT)(U/L) Mean 22.21 26.88 20.39 0.4768.sup.1) SD 13.90 18.82 13.41 Median 16.00 19.00 17.00 Min 7.00 10.00 8.00 Max 61.00 76.00 68.00 γ-GTP (U/L) Mean 18.18 33.58 24.12 0.0640.sup.1) SD 12.64 47.95 19.56 Median 14.00 19.00 16.00 Min 7.00 8.00 7.00 Max 61.00 270.00 81.00 p-value: ANOVA .sup.1)Kruskal-wallis test *p-value < 0.05, statistically significant

(51) The Alkaline phosphatase and γ-GTP items showed a statistically significant difference between three groups, as they were reduced in the high-dose group and low-dose group and they were increased in the control group, and a statistically significant difference before/after was shown in the high-dose group and low-dose group. ALT showed a statistically significant difference between three groups as it was reduced in the high-dose group and low-dose group and it was increased in the control group, and a statistically significant difference before/after was shown in the high-dose group. In this test, blood liver enzyme indexes reflecting liver function commonly showed the improved result after taking allulose, and nonalcoholic fatty liver disease is considered as another phenotype of metabolic syndrome caused by obesity. In conclusion, the results of analyzing the above body components showed that allulose inhibited body fat accumulation and muscle mass reduction.

Example 4: Safety Test

(52) In the physiochemical test for the safety, the low-dose group and high-dose group were examined as normal or clinically insignificant abnormal opinion in all items, and there was no statistically significant difference between the three groups. Twelve (12) specific items of the physiochemical test (ITT, Ears/Nose/Throat, Eye, /Head/Neck, Cardiovascular system, Abdomen, Respiratory system, Breast/Genitalia, Neurological system, Lymph nodes, Skin, Musculoskeletal, etc.) were classified into three (3) categories of “Normal, Abnormal (Not Clinically Significant), and Abnormal (Clinically Significant)”, and they were examined as normal or Abnormal (Not Clinically Significant) opinion in all the items, and there was no statistically significant difference between three groups.

(53) In addition, in the test result of vital signs, the average change in systolic blood pressure was reduced by 4.15±10.30 mmHg in the high-dose group and was reduced by 2.67±9.71 mmHg in the low-dose group, and was increased by 1.73±9.63 mmHg in the control group, at 8 weeks compared to the base time, which showed a statistically significant difference between the three groups (p-value=0.0349). The diastolic blood pressure and pulse rate did not showed statistically significant difference between three groups.

(54) Furthermore, the difference of the average change between the three groups at 12 weeks compared to the base time for the items was analyzed in the laboratory test. In the blood chemical test items, Alkaline phosphatase (p-value=<0.0001), ALT (SGPT) (p-value=0.0127), and γ-GTP (p-value=0.0043) showed statistically significant difference between the three groups. As the result of comparing the normality of the urine test, there was no clinically significant abnormal subject, and all items did not showed statistically significant difference between three groups.