Proton pump inhibitor, and supplement, food, beverage, or medicine comprising the proton pump inhibitor

Abstract

To provide a proton pump inhibitor for a gastrointestinal system, which contains polysaccharides extracted from an Aloe plant, and which can inhibit gastric acid production by means of the polysaccharides.SOLUTION: A proton pump inhibitor PPI inhibits the activity of a proton pump PP which exists in a secretory membrane S3 of parietal cells S2 of fundic glands S1 forming the stomach S0 within an organism. The proton pump PP is an H+, K+-ATPase, and it facilitates the release of H+ from the secretory membrane S3, on the basis of the binding of M3-R, H2-R, and G-R (GCCK2-R) in the parietal cells S2 with corresponding acetylcholine, histamine, and gastrin, thereby allowing the production of the gastric acid based on the released H+. The proton pump inhibitor PPI contains polysaccharides extracted from an Aloe plant. When the proton pump inhibitor PPI reaches the secretory membrane S3 from the external part of the organism, it inhibits the activity of H+, K+-ATPase to inhibit release of H+ (protons).SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a proton pump inhibitor that inhibits the activity of proton pumps in the gastrointestinal system, and to a supplement, food, beverage, or medicine containing the proton pump inhibitor. [Background technology]

[0002] Traditionally, the aloe plant has been recognized to have useful effects such as high healing power. It is believed that compositions derived from the aloe plant are involved in these effects. There are various views on the effectiveness of the aloe-derived composition with respect to its chemical and physical properties.

[0003] For example, the composition described in the following Patent Document 1 is an aloe-based composition containing aloe extract, Poria extract, and rosemary extract. The aloe extract contains polysaccharides, which enables the composition to regulate immune homeostasis. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. WO2022 / 015559 Summary of the Invention [Problem to be solved by the invention]

[0005] On the other hand, little research has been done on how compositions derived from aloe plants act on the mucous membranes of the body and how they exert their effects. As a result of intensive research, the present inventors have clarified the mechanism by which polysaccharides extracted from aloe plants act on the proton pump in the gastrointestinal system, and further discovered for the first time that they are effective as proton pump inhibitors.

[0006] In view of the above, the object of the present invention is to provide a gastrointestinal proton pump inhibitor, which contains polysaccharides extracted from an aloe plant and can suppress the production of gastric acid with the polysaccharides. The object of the present invention is also to provide a supplement, food, drink, or medicine containing the above-mentioned proton pump inhibitor, which can suppress the production of gastric acid with the polysaccharides. [Means for solving the problem]

[0007] The technical means of the present invention for solving this technical problem is characterized by the following points. The proton pump inhibitor of the present invention inhibits the activity of a proton pump present in the secretory membrane of the parietal cells of the fundus gland that constitutes the stomach in the body of an organism. The proton pump is H + ,K + -ATPase (proton / potassium-ATPase) in the parietal cells, 3 -R (muscarinic receptor), H 2 -R(H 2 receptor), GCCK 2 -R (gastrin receptor) binds to the corresponding acetylcholine, histamine, and gastrin, + (protons) from the secretory membrane, and the released H + The proton pump inhibitor comprises a polysaccharide extracted from an aloe plant, and when it reaches the secretory membrane in the body from outside the body of the organism, it is capable of generating gastric acid based on H + ,K + -ATPase (proton / potassium-ATPase) activity, + (proton) release is suppressed.

[0008] In the proton pump inhibitor of the present invention, at least -OH (hydroxyl group) and -COCH 3 (acetyl group), and in one sugar chain of the polysaccharide, the -COCH 3 The number of (acetyl groups) is less than the number of the -OH (hydroxyl groups).

[0009] In the proton pump inhibitor of the present invention, the mass of one sugar chain of the polysaccharide is in the range of 15 kDa (kilodalton) to 835 kDa (kilodalton).

[0010] In the proton pump inhibitor of the present invention, one sugar chain of the polysaccharide has at least a β(1,4)-mannose skeleton and a β(1,4)-glucose skeleton, and includes a sugar chain in which a compound of the following formula (1) or the following formula (2) is repeatedly bonded as a unit, and the β(1,4)-mannose skeleton has -COCH 3 (acetyl group). [ka] [ka] In this case, Ac in the above formula (1) and formula (2) is -COCH 3 (acetyl group).

[0011] The proton pump inhibitor of the present invention contains 40 to 95 parts by mass of a sugar chain in which the compound of formula (1) or formula (2) is repeatedly bonded as a unit relative to 100 parts by mass of the proton pump inhibitor.

[0012] The supplement, food, beverage, or medicine of the present invention contains the proton pump inhibitor described above. Effect of the Invention

[0013] According to the present invention, the activity of the proton pump can be inhibited by polysaccharides extracted from the aloe plant, and H + It can suppress the release of (protons), thereby suppressing the production of stomach acid. [Brief description of the drawings]

[0014] [Figure 1]FIG. 2 is a diagram for explaining the mechanism by which a proton pump inhibitor according to an embodiment of the present invention acts on a proton pump in the stomach in the body of an organism. [Diagram 2] FIG. 2 shows the mode of production and transport of each substance in parietal cells containing proton pumps, which are the targets of action of the proton pump inhibitors shown in FIG. 1. [Diagram 3] FIG. 2 shows the results of ATR-FTIR analysis of the composition contained in the proton pump inhibitor shown in FIG. 1. [Figure 4] FIG. 2 shows the results of 1H-NMR analysis of the composition contained in the proton pump inhibitor shown in FIG. 1. [Diagram 5] FIG. 2 shows the results of 13C-NMR analysis of the composition contained in the proton pump inhibitor shown in FIG. 1. [Figure 6] FIG. 2 shows the results of HPSEC-RI analysis of the compositions contained in the proton pump inhibitors shown in FIG. 1. [Figure 7] FIG. 2 shows the results of HPLC analysis of the compositions contained in the proton pump inhibitors shown in FIG. 1. [Figure 8] FIG. 2 shows the results of analyzing the compositions contained in the proton pump inhibitors shown in FIG. 1 by UV-Vis spectroscopy. [Figure 9] FIG. 2 shows the results of 1H-NMR analysis of the composition contained in the proton pump inhibitor shown in FIG. 1. [Figure 10] FIG. 2 shows the results of HPSEC-RI analysis of the compositions contained in the proton pump inhibitors shown in FIG. 1. [Figure 11] FIG. 2 shows the results of HPLC analysis of the compositions contained in the proton pump inhibitors shown in FIG. 1. [Figure 12] FIG. 2 shows the results of analyzing the compositions contained in the proton pump inhibitors shown in FIG. 1 by UV-Vis spectroscopy. [Figure 13] FIG. 2 shows the results of testing cell viability at each concentration when a composition containing the proton pump inhibitor shown in FIG. 1 was used. [Figure 14]A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 15] A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 16] A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 17] A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 18] A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 19] A diagram showing the test results of cell viability at each concentration when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 20] A diagram showing the results of analyzing the LDH release when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 21] A diagram showing the results of the protective effect of the composition against cell damage when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 22] A diagram showing the results of the protective effect of the composition against cell damage when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 23] A diagram showing the results of the protective effect of the composition against cell damage when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 24] A diagram showing the results of the protective effect of the composition against cell damage when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Diagram 25] A diagram showing the results of the protective effect of the composition against cell damage when using the composition contained in the proton pump inhibitor shown in Fig. 1. [Figure 26] FIG. 2 is a graph showing the results of measuring the weight of gastric juice when a composition containing the proton pump inhibitor shown in FIG. 1 was used. [Figure 27] FIG. 2 is a graph showing the results of measuring the pH of gastric juice when a composition containing the proton pump inhibitor shown in FIG. 1 was used. [Figure 28] FIG. 2 is a graph showing the results of measuring total acidity when a composition containing the proton pump inhibitor shown in FIG. 1 is used. [Figure 29] FIG. 2 is a graph showing the results of measuring the pH of gastric juice when a composition containing the proton pump inhibitor shown in FIG. 1 was used. [Diagram 30] FIG. 2 is a graph showing the results of measuring total acidity when a composition containing the proton pump inhibitor shown in FIG. 1 is used. [Diagram 31] FIG. 2 is a graph showing the results of measuring the damaged area of ​​the stomach when a composition containing the proton pump inhibitor shown in FIG. 1 was used. [Diagram 32] FIG. 2 is a graph showing the calculation results of the gastric lesion index when a composition contained in the proton pump inhibitor shown in FIG. 1 was used. [Diagram 33] FIG. 2 is a diagram showing a summary of the effects when a composition containing the proton pump inhibitor shown in FIG. 1 is used. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0016] As shown in Fig. 1, the proton pump inhibitor PPI according to an embodiment of the present invention is a composition that inhibits the activity of the proton pump PP in the stomach S0. The stomach S0 is the stomach S0 in the body of an organism and is composed of a fundus gland S1. Parietal cells S2 of the fundus gland S1 have a secretory membrane S3.

[0017] The proton pump PP is an enzyme present in the secretory membrane S3. + ,K +The proton pump PP is a proton / potassium-ATPase. + (protons) from the secretory membrane S3.

[0018] As shown in Figure 2, more specifically, inside the parietal cell S2, water H 2 O is H + , O.H. - (hydroxide ion). + is released into the stomach S0 lumen by the proton pump PP. - Carbon dioxide CO inside the parietal cells S2 2 Combines with HCO 3 - (bicarbonate ion) is produced.

[0019] HCO produced 3 - is Cl - ,HCO 3 - By exchange transporters, Cl is released into the blood outside the parietal cell S2. - ,HCO 3 - The Cl in blood is transported by exchange transporters. - (Chloride ions) are taken up into the interior of the parietal cell S2.

[0020] On the other hand, in the proton pump PP, H + In response to the release of + (potassium ions) are taken into the parietal cell S2. K + is transported to the bloodstream, while Cl - is transported to the lumen of the stomach S0. H released by the proton pump PP + and transported Cl - Based on this, HCl, which is gastric acid, can be produced.

[0021] As shown in Fig. 1, the parietal cells S2 contain M 3-R (muscarinic receptor), H 2 -R(H 2 receptor), and GR (GCCK 2 -R (gastrin receptor)) are present in parietal cells S2. 3 -R, H 2 -R, GCCK 2 -R binds to the corresponding acetylcholine, histamine, and gastrin to activate the proton pump PP, H + ,K + -ATPase is activated.

[0022] More specifically, M 3 When acetylcholine binds to Ca + Increases the concentration of calcium ions. 2 When -R binds to histamine, it increases the concentration of cAMP (cyclic adenosine monophosphate). 2 When bound to gastrin, -R induces Ca + Increases the concentration of cAMP and Ca + In response to increases in the concentration of each of these, protein kinases are activated, resulting in the formation of H + ,K + -ATPase is activated.

[0023] Here, when the proton pump inhibitor PPI reaches the secretory membrane S3 (parietal cell S2) inside the body of an organism, the proton pump inhibitor PPI acts as a H + ,K + More specifically, proton pump inhibitors (PPIs) that reach the parietal cells S2 are activated by gastric acid and are converted into H + ,K + -Binds to ATPase.

[0024] This allows H + ,K + The SH group of H-ATPase is modified, and the enzyme activity is suppressed. + ,K + -ATPase, H +In this way, the action of the proton pump inhibitors (PPIs) mentioned above inhibits the production of gastric acid, which is caused by the binding of acetylcholine, histamine, and gastrin.

[0025] The proton pump inhibitor PPI according to the embodiment of the present invention contains polysaccharides extracted from an aloe plant. The aloe plant used for extraction may be any species of aloe plant. For example, one or more of Aloe Vera, Aloe arborescens, Aloe Barbadense, Aloe Ferox, etc. may be selected and used for extraction. The extraction of polysaccharides from an aloe plant may be carried out through a process such as extraction with a solvent or concentration by chromatography.

[0026] The polysaccharide contained in the proton pump inhibitor PPI may have any chemical structure and molecular weight as long as it has a sugar chain based on a sugar skeleton such as glucose, mannose, or galactose. One sugar chain of the polysaccharide has at least -OH (hydroxyl group) and -COCH 3 (acetyl group).

[0027] More specifically, -OH may be contained in each of the glucose backbone and the mannose backbone that constitute the sugar chain. 3 may be contained in the mannose skeleton that constitutes the glycan. In one glycan of the polysaccharide contained in the proton pump inhibitor PPI, -COCH 3 For example, in one glycan, one, two, or three of the -OH groups in the mannose backbone may be -COCH 3 and the -OH of the glucose backbone is maintained, resulting in -COCH 3 may be less than the number of -OH.

[0028] In addition, the degree of acetylation in one sugar chain of a polysaccharide is, for example, in the range of 30% to 70%. More specifically, the degree of acetylation may be, for example, in the range of 30% to 45%, 45% to 55%, or 55% to 70%. Here, the degree of acetylation is determined by the degree of acetylation of -OH of the mannose skeleton to -COCH 3 For example, the ratio of -OH in the mannose skeleton to -COCH 3 If the -OH of the mannose backbone is not replaced by -COCH, the degree of acetylation is zero. 3 When all of the acetyl groups are replaced by , the degree of acetylation is 100%.

[0029] -COCH contained in polysaccharides 3 -COCH is thought to play an important role in physiological activity. 3 By containing, for example, it is expected that the antioxidant effect and the immune regulation function will be improved. In particular, in one sugar chain of a polysaccharide, as described above, -COCH 3 By adjusting the number of H + ,K + This makes it possible to realize a proton pump inhibitor that is more suitable for inhibiting the activity of .BETA.-ATPase.

[0030] The mass of one sugar chain of the polysaccharide contained in the proton pump inhibitor PPI is, for example, in the range of 15 kDa (kilodalton) to 835 kDa (kilodalton). The mass of the sugar chain is preferably small from the viewpoint of water solubility and ease of diffusion into a medium, and may be, for example, in the range of 15 kDa to 100 kDa, 100 kDa to 200 kDa, 200 kDa to 300 kDa, 300 kDa to 400 kDa, 400 kDa to 500 kDa, 500 kDa to 600 kDa, 600 kDa to 700 kDa, or 700 kDa to 835 kDa.

[0031] One sugar chain of the polysaccharide contained in the proton pump inhibitor PPI may have at least a β(1,4)-Mannose (mannose) skeleton and a β(1,4)-Glucose (glucose) skeleton. One sugar chain of the polysaccharide contains a sugar chain in which the compounds of the following formula (1) or the following formula (2) are repeatedly bonded as units. In the following formula (1) and formula (2), only the -OH corresponding to the C at the 3rd position in the β(1,4)-Mannose skeleton on the left end is -COCH 3 (acetyl group) is contained. Ac in the above formula (1) and formula (2) represents -COCH 3 . n in the above formula (1) and formula (2) represents the degree of polymerization. The degree of polymerization n may be adjusted so that the mass of the sugar chain changes within the above-mentioned range.

[0032]

Chemical formula

[0033]

Chemical formula

[0034] The compound shown in the above formula (1) has four saccharide skeletons and is composed of a β(1,4)-Mannose skeleton, a β(1,4)-Mannose skeleton, a β(1,4)-Glucose skeleton, and a β(1,4)-Mannose skeleton from left to right.

[0035] Each Mannose skeleton has a cyclic structure. The cyclic structure is composed of 5 C (carbons) and 1 O (oxygen). Starting from the O, numbers 1, 2, 3, 4, 5 are assigned to the 5 C in a clockwise direction on the paper surface. Also, a number 6 is assigned to the C that is bonded to the C at the 5th position and is outside the cyclic structure.

[0036] In the β(1,4)-Mannose skeleton at the left end, only the -OH corresponding to the C at the 3rd position is -COCH 3In the second β(1,4)-Mannose skeleton from the left, only the -OH corresponding to the 2nd C is replaced by -COCH 3 In the β(1,4)-Mannose skeleton on the right, only the -OH corresponding to the 3rd C is replaced by -COCH 3 has been replaced by.

[0037] The compound shown in the above formula (2) is obtained by adding an α(1,6)-galactose skeleton to the compound shown in the above formula (1). More specifically, one α(1,6)-galactose skeleton is connected to the C at the 6th position of the second β(1,4)-mannose skeleton from the left. The compound shown in the above formula (2) differs from the compound shown in the above formula (1) only in this respect.

[0038] The proton pump inhibitor PPI according to the embodiment of the present invention contains 40 to 95 parts by mass of a sugar chain in which the compound of formula (1) or formula (2) is repeatedly bonded as a unit, relative to 100 parts by mass of the proton pump inhibitor. The content of the sugar chain can be adjusted depending on the production process and other additives added to the proton pump inhibitor PPI, and may be, for example, in the range of 40 to 50 parts by weight, 50 to 60 parts by weight, 60 to 70 parts by weight, 70 to 80 parts by weight, 80 to 90 parts by weight, or 90 to 95 parts by weight.

[0039] The proton pump inhibitor PPI according to the embodiment of the present invention may be contained in a supplement, food, beverage, or medicine. The food containing the proton pump inhibitor PPI may be configured as a health food, functional food, etc. The beverage containing the proton pump inhibitor PPI may be configured as a health drink, functional drink, etc. The supplement or medicine containing the proton pump inhibitor PPI may be in any form such as a solid, liquid, gel, etc., and more specifically, may be an oral tablet, a granule contained in a capsule, or a liquid or gel sprayed by an inhaler.

[0040] <Analysis of Proton Pump Inhibitors> The components contained in the proton pump inhibitor PPI will be described with reference to Figs. 3 to 12. The components were analyzed by analyzing the molecular structure, molecular weight, and amount of acetyl groups of the compounds. FTIR and NMR were used for the analysis of molecular structure. HPSEC-RI and HPLC were used for the analysis of molecular weight. UV-Vis spectroscopy was used for the analysis of the amount of acetyl groups.

[0041] The proton pump inhibitors (PPIs) to be analyzed include compositions obtained by purifying extracts from Aloe vera leaves. The compositions were obtained by various extraction methods, and two types of compositions were analyzed. The two types of compositions are referred to as composition A50 and composition I50, respectively. Composition A50 corresponds to the compound shown in formula (1) above, and composition I50 corresponds to the compound shown in formula (2) above.

[0042] Figure 3 shows the results of ATR-FTIR analysis of composition A50. The four main functional groups (OH, CO, CH 3 , and COO) were confirmed. In the spectrum shown in FIG. -1 , ~2924cm -1 , ~1734cm -1 , 1236cm -1 The peaks are the stretching vibrations of -OH (hydroxyl group), -CH 2 and -CH 3 (aliphatic group), stretching vibration of C=O in -COO (carbonyl group) of polysaccharides, -COCH of polysaccharides 3 These correspond to the stretching vibration of CO in the acetyl group.

[0043] FIG. 4 shows composition A50. 14 shows the results of analysis by H-NMR. In the spectrum shown in FIG. 4, three signals at 2.10 ppm, 2.06 ppm, and 2.02 ppm correspond to methyl groups. This indicates that the acetyl groups are located at different positions. The signal at 2.00 ppm corresponds to O-acetyl, and the O-acetyl content in composition A50 was 0.75.

[0044] FIG. 5 shows composition A50. 13 5 shows the results of analysis by C-NMR. In the spectrum shown in FIG. 5, the three signals at 20.54 ppm, 20.27 ppm, and 20.06 ppm correspond to methyl groups. The three signals at 173.93 ppm, 173.42 ppm, and 12.58 ppm correspond to carboxyl carbons of acetyl groups. This indicates that composition A50 contains acetylated mannan (e.g., Acemannan).

[0045] FIG. 6 shows the results of analyzing composition A50 by HPSEC-RI (high performance size exclusion chromatography-differential refractive index detection). The relative molecular weight of the sample was obtained based on the largest peak point detected within the trend line range. The trend line was obtained by plotting using a pull line standard. In FIG. 6, the relative molecular weight of the first peak on the left side was 180 kDa (average value within the range of the first peak). The second peak on the right side outside the trend line at 45 to 50 minutes indicates the presence of a molecular weight smaller than 5 kDa. The results of analyzing composition A50 by HPLC (high performance liquid chromatography) are also shown in FIG. 7.

[0046] FIG. 8 shows the results of analyzing composition A50 by UV-Vis spectroscopy. First, the acetyl group of composition A50 was reacted with hydroxylamine to form acetohydroxamic acid. The resulting compound was then reacted with ferric chloride to form a ferric-acetohydroxamic complex. The ferric-acetohydroxamic complex is red and has an absorption peak at a wavelength of 540 nm. The O-acetyl content in the ferric-acetohydroxamic complex can be determined by the absorbance at that wavelength. Therefore, the absorbance of the complex at a wavelength of 540 nm was detected. The detected absorbance was compared with a calibration curve (reference standard: acetylcholine chloride≧99% (TLC)).

[0047] The detailed method is as follows. First, 0.5 mL of each of the standard solution, the diluted sample, and water (blank group) were taken into 20 mL glass vials. Next, 0.5 mL of 4 M HCl was added to the blank group and shaken. Next, 1 mL of alkaline hydroxylamine hydrochloride solution was added to each glass vial, shaken, and left at room temperature for 4 minutes. Next, 0.5 mL of 4 M HCl was added to each glass vial except the blank group, shaken, and adjusted to a pH of 1.2 ± 0.2. Next, 0.5 mL of 0.62 M ferric chloride was added to each glass vial and stirred with a vortex mixer. At this time, the solution turned reddish orange. After removing air bubbles from the samples using an ultrasonic device, the absorbance at 540 nm of the standard solution and the sample was measured. The O-acetyl content of the sample was calculated according to the absorbance value of the standard solution. The calculation results are shown in the table at the bottom of Figure 8.

[0048] FIG. 9 shows composition I50. 1 9 shows the results of H-NMR analysis. In the spectrum shown in FIG. 9, the signal at 2.00 ppm corresponds to O-acetyl, and the O-acetyl content in composition I50 was 1.62.

[0049] Figure 10 shows the result of analyzing Composition I50 by HPSEC-RI (high performance size exclusion chromatography - refractive index detection). Based on the largest peak point detected within the range of the trend line, the relative molecular weight of the sample was obtained. The said trend line was obtained by plotting using pullulan standards. In Figure 10, the relative molecular weight at the first peak on the left side was 37 kDa (average value within the range of the first peak). The second peak on the right side outside the trend line at 458 minutes indicates the presence of a molecular weight smaller than 5 kDa. The result of analyzing Composition I50 by HPLC (high performance liquid chromatography) is also shown in Figure 11.

[0050] Figure 12 shows the result of analyzing Composition I50 by UV-Vis spectroscopy. The analysis process is the same as described above. The calculation results of the O-acetyl content of the sample are shown in the table at the lower part of Figure 12.

[0051] <Treatment and / or prevention of IBD (in vitro test)> RGM1 (rat gastric mucosa) cells were cultured in DMEM / F12 (Corning, USA) supplemented with 10% (v / v) fetal bovine serum (FBS, HyClone, USA) and 1% (v / v) antibiotic-antimycotic (Gibco, Thermo Fisher Scientific, USA). The cells were incubated at 37 °C in a 5% CO 2 atmosphere. Before each experiment, the culture was washed with phosphate buffered saline (PBS, pH 7.4, PROTECH, Taiwan) and detached with 0.05% trypsin-EDTA solution (Gibco, Thermo Fisher Scientific, USA). Then, centrifugation (Hermle Z 36 HK, Germany) was performed at 1700 rpm for 5 minutes, followed by resuspension in complete medium.

[0052] The details of the in vitro cytotoxicity assay are described below. A total of 1×104 cells / well were seeded in a 96-well plate and incubated for 12 hours for cell attachment. 50 mg / mL stock solutions of compositions with different degrees of acetylation (e.g., acemannan) were prepared directly in the medium. The degrees of acetylation to be adjusted were 10%, 42%, and 50%. Excipients were used in the 10% formulation. The concentrations of excipients were calculated corresponding to the 10% formulation. Cells were treated with different concentrations of compositions (e.g., acemannan) and incubated for 24 and 48 hours. The concentrations used for treatment were 1000 mg / mL, 500 mg / mL, 200 mg / mL, 100 mg / mL, and 50 mg / mL. After treatment, 10 mL of CCK-8 solution (Dojindo Laboratories, Japan) was added to each well and the plate was incubated for another hour. Absorbance readings at 460 nm were performed using a microplate reader (EPOCH2, BioTek Instruments, Inc., USA). Data are presented as mean ± SD, and statistical significance was analyzed by Student's t-test. A p-value of 0.05 was considered statistically significant. Photomicrograph images of cells treated with different concentrations of compositions (e.g., acemannan) were taken at 10x and 20x magnifications using a Zeiss Axio Vert.A1, Germany.

[0053] Samples with 10%, 42%, and 50% acetylation were weighed as necessary and dissolved directly in serum-free medium. The sample with 10% acetylation degree can be composed of a sample with 40% acemannan or 50% acetylation degree and a pure excipient (e.g., cornstarch). That is, a sample with 10% acetylation degree can be prepared by simply adding an excipient to dilute a sample with 50% acetylation degree. In this case, the degree of acetylation was actually 10% to 12%. The sample for the cytotoxicity test is a powder rather than a tablet, but the recommended intake amount when formed into a tablet is 1 to 2 tablets / day for adults. The mass of the tablet may be, for example, 100 mg (composition (e.g., acemannan) + excipient).

[0054] Figures 13 to 19 show the test results. In these figures, A10, A42, and A50 correspond to samples with acetylation degrees of 10%, 42%, and 50%, respectively. When cells were treated with A10 and the activator, significant cell proliferation was observed after 48 hours. When A42 and A50 were used, cell proliferation decreased after 24 hours of incubation, and increased after 48 hours of incubation (up to 500 mg / mL). The morphology and viability of RGM1 cells under different treatment conditions were also evaluated by microscope, but the cell morphology did not show any significant change under all treatment conditions. Details of the evaluation by microscope are omitted.

[0055] Figure 20 shows the results of analyzing the LDH release of composition A50. As a pretreatment, RGM1 was treated with composition A50 for 1 hour, then washed and replenished with fresh complete medium. RGM1 was treated with composition A50 for 24 or 48 hours by co-incubation. The results showed that there was no leakage of LDH from RGM1 treated with composition A50 (1 μg / mL) for 48 hours, and no cytotoxicity.

[0056] 21 to 25 show the results of the protective effect of Composition A50 against cell damage. The cell damage in this evaluation is caused by indomethacin.

[0057] From the above results, the edible composition (e.g., acemannan) has the advantages of attenuating the digestive process, increasing satiety, and delaying sugar release from starch for gastrointestinal health. In the colon, the composition (e.g., acemannan) is converted by microorganisms into short-chain fatty acids, providing a prebiotic effect to the host. As a soluble dietary fiber, the composition (e.g., acemannan) has the effects of increasing mineral absorption, suppressing obesity, suppressing inflammation and related comorbidities (colon cancer, IBD, etc.), improving laxity (defecation), and reducing chronic systemic inflammation.

[0058] <Improvement of mucosal protection (in vivo test)> A small-scale pilot study was conducted to evaluate the mucosal protective function. The study followed the government regulation on the evaluation method for improving gastrointestinal function. An evaluation model and active dose were established and the evaluation was carried out.

[0059] Eleven mice were used for the evaluation. The mice used were 6-week-old male BALB / cAnNCrlBltw mice weighing 19-22 g. Three of them were in the Sham group, three in the Vehicle group, and five in the Acemannan group.

[0060] The mice were kept in an environment with a temperature of 22±3°C, a relative humidity of 50±20%, and a light / dark cycle of 12 hours light and 12 hours dark. The mice were kept on absorbent corn cob beds (10054, Andersons, USA), with sterilized feed (MFG, Oriental Yeast Co., Ltd., Japan), and in individually ventilated cages (IVC). Sterilized RO water was constantly supplied to the cages.

[0061] As shown in Table 1, the test referred to the method for evaluating the improvement of gastrointestinal function in health foods (Ministry of Health and Welfare Notification No. 0920401629). Oral administration was performed at a dose of 500 mg / kg (BID) to mice.

[0062] [Table 1]

[0063] a: The drug was administered twice a day at 6-hour intervals for 7 consecutive days. Gastric acid secretion was measured on the morning of the 8th day after the final administration. b: Oral administration

[0064] A homogenous solution of the substance was prepared using sterile water for injection. The route and method of administration were in accordance with the "Standard Operating Procedure for Sample Administration" (SHGLP-SOP-IA-020). On the day of administration, an amount of substance corresponding to the weight of the mouse was taken using a sterile syringe and orally administered to the mouse through a plastic feeding needle. The amount administered to each mouse was calculated based on the mouse's weight (weight before the substance was administered on the first day). The day the substance was first administered was defined as day 1. The weight of the mice was measured on days 1 and 8 according to the "Standard Operating Procedure for Laboratory Animal Care" (SHGLP-SOP-IA-003).

[0065] After substance administration was started, the "Standard Operating Procedure for Animal Ward Rounds" (SHGLP-SOP-IA-009) was followed. Clinical signs of mice were observed and recorded at least once daily throughout the study period. Mice were monitored for survival twice daily with at least 6-h intervals.

[0066] Evaluation was performed using Shay's ulcer method. Mice in the Sham, Vehicle and Acemannan groups were fasted for 24 hours. The pylorus of the stomach was ligated and then sutured for recovery. After 4 hours, the stomach was removed from the mice. Gastric juice was collected, quantified and titrated with 0.01N NaOH solution. The change in total acidity between the experimental and control groups was calculated. p<0.05 indicates that the composition (e.g., acemannan) has the function of protecting the gastric mucosa.

[0067] Data are expressed as mean ± standard deviation (SD). Data were analyzed using t-test, and a statistically significant difference was determined compared to the Sham group using a significance criterion of ≦0.05.

[0068] Table 2 shows a summary of the body weight, clinical symptoms, and autopsy findings of the animals on days 1 and 8. As a result, no animals died during the experiment. No significant difference in weight was observed before and after administration of the substance. No adverse clinical symptoms were observed during the test period (days 1 to 8). No abnormal findings were observed at autopsy. Based on these results, the mice did not lose weight or show any observable side effects even after continuous administration of Acemannan (500 mg / kg, BID) for 8 days. In addition, no gross abnormalities were observed in the organs.

[0069] [Table 2]

[0070] Table 3 and Figures 26 to 28 show the results of gastric juice weight, gastric juice pH measurement, and total acidity measurement. On the morning of the 8th day, Acemannan and Vehicle were administered, and then Shay's ulcer method was performed. After 24 hours of fasting, the stomach was ligated at the junction of the pylorus and jejunum. Four hours later, the mice were sacrificed and gastric juice was collected.

[0071] [Table 3]

[0072] In the above table, *p<0.05 compared to G1.

[0073] No significant differences were observed in gastric juice weight and pH (G1: Sham, G2: Vehicle (sterile water), G3: Acemannan 500mg / kg, BID). However, the total acidity of G3 group (0.390±0.122) was significantly lower than that of G1 group (0.660±0.181). The total acidity of G2 group (0.620±0.167) given sterile water did not show a significant difference from that of G1 group. Based on these data, it can be concluded that after 8 days of continuous administration of Acemannan (500mg / kg, BID), no statistically significant differences were observed in gastric juice weight and pH when compared with the Sham-fed G1 group using the Shay's ulcer method. On the other hand, the total acidity showed a significant decrease.

[0074] A test was carried out on male Sprague Dawley rats (50 rats in total, 8 weeks old) to examine the gastroprotective effect of compositions A10 and A50. After an adaptation period, the rats were randomly divided into 10 groups (n=5 / group). The feeding pattern for each group is shown below. (1) NC1 group: underwent sham surgery and were provided with distilled water at 10 mL / kg BW / day (2) NC2 group: Provided with distilled water at 10 mL / kg BW / day (3) NC3 group: 335 mg of excipient (4) PC Group: Clinical Drug 5-Aminosalicylic Acid (5) L-A10 group: composition A10, 100 mg / daily dose for humans (6) M-A10 group: Composition A10, 300 mg / daily dose in humans (7) H-A10 group: Composition A10, 500 mg / daily human dose (8) L-A50 group: composition A50, 33 mg / daily human dose (9) M-A50 group: composition A50, 100 mg / daily dose for humans (10) H-A50 group: Composition A50, 167 mg / daily human dose

[0075] All rats were preconditioned for 1 week, and 50 SD rats were allowed to acclimate to the environment before being grouped as described above. The rats were housed in plastic cages, with one or two per cage. First, the rats were assigned identification numbers, and the rats were weighed and grouped to balance the weight. Throughout the experiment, reverse osmosis purified water was provided as drinking water, and the rats were allowed to have water ad libitum. The rats were also provided with standard animal chow (Chow 5001). In this experiment, the rats were weighed regularly to monitor their health and the dosage of the test sample for each rat was determined based on their weight. The test samples were weighed daily according to the dosage required for each group. The samples were then dissolved in sterile water and orally administered to the rats using a feeding tube for 7 days. Clinical observations were performed daily during the experiment to record any abnormal clinical symptoms or death. The rats' weights were recorded every 2 days and monitored for changes. The rats' food intake was recorded every 3 days, and the rats' water intake was recorded daily.

[0076] The rats were continuously administered supplements containing compositions A10 and A50 for 7 days, and then fasted for 24 hours. The rats' abdomens were shaved along the midline of the xiphoid process, and a 1 cm incision was made along the midline to perform pylorus ligation. The incision was then sutured and the rats were allowed to recover. After 4 hours, the abdomen was incised along the midline to expose the stomach. The stomach was removed, and gastric juice was collected and quantified. The collected gastric juice was titrated with 0.1N NaOH solution. Total gastric acidity was defined as follows: Total gastric acidity (mEq / L) = gastric acid amount (L) x added sodium hydroxide (NaOH) amount (L) x 0.1N (NaOH solution titration) x 1000.

[0077] After washing the stomach with saline, the stomach was photographed in a flat position and the images were analyzed using the image analysis software ImageJ. The images of the gastric mucosa were converted to black and white and quantified by adjusting the appropriate image threshold. The damaged area was measured in mm 2 The gastric lesion index was calculated using the following formula: Gastric lesion index = gastric damage area (mm 2 ) / total gastric mucosa area (mm 2) × 100. The results are shown in Figures 29 to 33.

[0078] All values ​​are presented as mean ± SD. Sample size was 5 rats per group. One-way analysis of variance (ANOVA) was performed using the SAS statistical software package. Duncan's test was used for post-hoc comparison of group differences, and a level of p<0.05 was considered statistically significant.

[0079] No significant differences were observed in body weight, food, or water intake between the groups. The results showed that continuous administration of high, medium, and low doses of Compositions A10 and A50 supplements for 7 days significantly reduced total gastric acidity, gastric damage area, and gastric lesion index (p<0.05). Therefore, this experiment confirmed the gastroprotective effect of the test substances "A10 supplement" and "A50 supplement" on the gastric mucosa.

[0080] <Effects of the embodiment> As described above, the proton pump inhibitor PPI according to the embodiment of the present invention inhibits the activity of the proton pump PP present in the secretory membrane S3 of the parietal cell S2 of the fundus gland S1 constituting the stomach S0 in the body of an organism. + ,K + -ATPase (proton / potassium-ATPase) in parietal cells S2 3 -R (muscarinic receptor), H 2 -R(H 2 receptor), GR(GCCK 2 -R (gastrin receptor)) binds to the corresponding acetylcholine, histamine, and gastrin, + (protons) from the secretory membrane S2, and the released H + Proton pump inhibitors (PPIs) contain polysaccharides extracted from the aloe plant, and when they reach the secretory membrane S2 inside the body, they can generate gastric acid based on H + ,K + -ATPase (proton / potassium-ATPase) activity,+ (proton) release is suppressed.

[0081] According to this, polysaccharides extracted from aloe plants can inhibit the activity of the proton pump PP, and H + It can suppress the release of (protons), thereby suppressing the production of stomach acid.

[0082] In the proton pump inhibitor PPI according to the embodiment of the present invention, at least -OH (hydroxyl group) and -COCH 3 (acetyl group), and in one sugar chain of a polysaccharide, -COCH 3 The number of -acetyl groups is less than the number of -OH (hydroxyl groups).

[0083] According to this, in polysaccharides, -COCH 3 By appropriately adjusting the number of -acetyl groups and -OH (hydroxyl groups), the polysaccharide can be made to act effectively on the gastrointestinal system.

[0084] In the proton pump inhibitor PPI according to an embodiment of the present invention, the mass of one sugar chain of the polysaccharide is in the range of 15 kDa (kilodalton) to 835 kDa (kilodalton).

[0085] This allows the molecular weight of the polysaccharide to be relatively small, and the ease of diffusion into a medium (particularly water solubility) to be improved, which makes it easier to purify the polysaccharide, and allows a relatively large amount of the polysaccharide to be uniformly contained in supplements, foods, beverages, medicines, etc.

[0086] In the proton pump inhibitor PPI according to the embodiment of the present invention, one sugar chain of the polysaccharide has at least a β(1,4)-mannose backbone and a β(1,4)-glucose backbone, and includes a sugar chain in which the compound of the above formula (1) or the above formula (2) is repeatedly bonded as a unit, and the β(1,4)-mannose backbone is linked to -COCH 3(acetyl group).

[0087] According to this, by giving the polysaccharide a specific chemical structure, the polysaccharide can act more effectively on the gastrointestinal system.

[0088] The proton pump inhibitor PPI according to an embodiment of the present invention contains 40 to 95 parts by mass of a sugar chain in which the compound of formula (1) or formula (2) is repeatedly bonded as a unit relative to 100 parts by mass of the proton pump inhibitor.

[0089] According to this, when administering the proton pump inhibitor PPI, the content of polysaccharides can be adjusted in advance. In addition, when the proton pump inhibitor PPI (polysaccharides) is contained in supplements, foods, beverages, medicines, etc., the amount of polysaccharides can be appropriately adjusted according to the object to be contained. For example, by improving the content of polysaccharides, efficient intake is possible. On the other hand, by suppressing the content of polysaccharides, the flavor of foods, beverages, etc. can be prevented from being impaired.

[0090] The proton pump inhibitor PPI according to embodiments of the present invention may be included in a supplement, food, beverage, or medicine.

[0091] This allows the proton pump inhibitor PPI to be easily ingested.

[0092] The above-mentioned embodiments disclosed herein are illustrative in all respects and should not be considered as limiting. The scope of the present invention is defined by the claims, not the above description, and is intended to include all modifications within the meaning and scope of the claims. [Explanation of symbols]

[0093] S0…stomach PP...Proton pump PPI: Proton pump inhibitor

Claims

1. The fundus glands that make up the stomach in an organism's body Located in the secretory membrane of parietal cells Proton Pump In a proton pump inhibitor which inhibits the activity of The proton pump comprises: H ＋ , K ＋ -ATPase (proton / potassium-ATPase), M in the parietal cells ３ -R (muscarinic receptor), H ２ -R(H ２ receptor), GCCK ２ Based on the fact that the gastrin receptor (H-R) binds to the corresponding acetylcholine, histamine, and gastrin, ＋ (protons) from the secretory membrane, and the released H ＋ (protons) based stomach acid can be generated, The proton pump inhibitor is The polysaccharides are extracted from an aloe plant, and when they reach the secretory membrane inside the body of the organism from outside the body of the organism, the H ＋ , K ＋ -ATPase (proton / potassium-ATPase) activity to inhibit the H ＋ Suppresses the release of (protons) Proton pump inhibitors

2. 2. The proton pump inhibitor of claim 1, One sugar chain of the polysaccharide is At least -OH (hydroxyl group) and -COCH ３ (acetyl group), In one sugar chain of the polysaccharide, The -COCH ３ The number of (acetyl groups) is less than the number of the —OH (hydroxyl groups). Proton pump inhibitors.

3. 3. The proton pump inhibitor according to claim 2, The mass of one sugar chain of the polysaccharide is In the range of 15 kDa (kilodaltons) to 835 kDa (kilodaltons) Proton pump inhibitors.

4. The proton pump inhibitor according to any one of claims 1 to 3, One sugar chain of the polysaccharide is The present invention relates to a sugar chain having at least a β(1,4)-mannose skeleton and a β(1,4)-glucose skeleton, in which a compound represented by the following formula (1) or the following formula (2) is repeatedly bonded as a unit: The β(1,4)-mannose backbone is -COCH ３ (acetyl group) is included Proton pump inhibitors. 【Chemistry 1】 【Chemistry 2】 In the above formula (1) and formula (2), Ac is -COCH ３ (acetyl group).

5. 5. The proton pump inhibitor according to claim 4, The proton pump inhibitor contains 40 to 95 parts by mass of a sugar chain in which the compound of formula (1) or formula (2) is repeatedly bonded as a unit, relative to 100 parts by mass of the proton pump inhibitor. Proton pump inhibitors.

6. A supplement, food, beverage, or medicine comprising the proton pump inhibitor according to claim 4.

7. A supplement, food, beverage, or medicine comprising the proton pump inhibitor according to any one of claims 1 to 5.

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