JPS647972B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a digestive enzyme-containing gastrointestinal drug containing two or more starch-digesting enzymes with different enzymatic actions, and more specifically, a gastrointestinal drug containing two or more starch-digesting enzymes with different enzymatic actions. A gastrointestinal drug containing digestive enzymes that is extremely effective in rapidly and sufficiently decomposing starch through the synergistic effect of these enzymes, and rapidly alleviating unpleasant gastrointestinal symptoms caused by overeating, indigestion, etc. Regarding. Although dietary habits have changed recently, carbohydrate intake is still overwhelmingly higher than protein and fat, and starch-digesting enzymes play an extremely important role in alleviating unpleasant gastrointestinal symptoms. big. Currently, there are many gastrointestinal medicines containing starch digestive enzymes on the market, but the activity of the digestive enzymes is determined by measuring the saccharifying power based on the total amount of reducing ends, and the activity of these digestive enzymes is determined by measuring the saccharifying power from the total amount of reducing ends. It has not been evaluated from the perspective of decomposition. The present inventors have established a method for separating and quantifying oligosaccharides such as maltose, maltotriose, and maltotetraose produced by enzymatic decomposition of starch using high-performance liquid chromatography. By using the reducing end measurement method in combination, we succeeded in accurately understanding the actions of various starch-degrading enzymes, and based on this, we believe that by combining various starch-digesting enzymes with different actions in appropriate proportions, we can improve the efficiency of individual digestion. The present inventors have discovered that an effect greater than the sum of the effects expected from enzymes, that is, a synergistic effect can be obtained, and have completed the present invention. That is, the present invention provides a digestive enzyme-containing gastrointestinal medicine containing two or more types of starch digestive enzymes having different enzymatic actions. A preferred embodiment of the present invention is to mix liquefied α-amylase and saccharified α-amylase in an appropriate ratio. As is well known, α-amylase randomly decomposes α-1,4-bonds in starch molecules, so starch is decomposed into dextrin acid and rapidly liquefied, but it is difficult to decompose into maltose. On the other hand, β-amylase separates maltose molecules from the non-reducing ends of starch molecules in an orderly manner, so saccharification progresses steadily although the liquefaction rate is slow. Liquefaction here refers to the decomposition of starch into dextrin and solubilization, and saccharification refers to the decomposition of starch into oligosaccharides.From a physiological perspective, the former relieves abdominal bloating. is,
The latter may be considered to enable absorption from the gastrointestinal tract. As mentioned above, the main action of α-amylase is to break down starch into dextrin. In other words, even if α-amylase acts on starch, only a small amount of oligosaccharide is produced. However, it has recently become known that certain α-amylases degrade starch to yield relatively large amounts of oligosaccharides. Therefore, those with a low oligosaccharide production rate are called liquefied α-amylases, and those with a high oligosaccharide production rate are called saccharification α-amylases. In this way, the existence of saccharification-type α-amylase itself is known, but the various α-amylase products actually used have not been analyzed in detail to determine whether they are liquefied or saccharified. There are no examples. The present inventors analyzed various commercially available α-amylases using a method for separating and quantifying oligosaccharides using high-performance liquid chromatography, which will be described in detail later, and classified them into liquefaction type, saccharification type, and intermediate type α-amylase. We were able to. Based on the results, starch was degraded using a sample containing an appropriate mixture of liquefied α-amylase and saccharified α-amylase, and the amount of maltose and maltotriose produced was determined using the separation quantitative method described in detail later. However, it was found that these blended samples exhibited a significant synergistic effect. This synergistic effect is due to the fact that liquefied α-amylase rapidly breaks down starch into low-molecular-weight dextrins, while saccharified α-amylase degrades low-molecular-weight dextrins and starch that are degraded by liquefied α-amylase. In order to decompose all non-reducing ends,
This seems to be due to the fact that the amount of maltose and maltotriose produced increases at an accelerated rate compared to when liquefied α-amylase or saccharified α-amylase is used alone. For example, when Diasmen SS is used as the liquefied α-amylase and biodiastase is used as the saccharified α-amylase, the ratio of the former to the latter is the reducing sugar activity ratio (the amount of reducing sugar produced when the enzyme acts on the substrate). It was found that a high synergistic effect can be obtained when the ratio (when evaluating enzyme activity) is 4:1 to 1:3. On the other hand, as mentioned above, unpleasant symptoms such as abdominal bloating and heaviness in the stomach are quickly relieved by the action of α-amylase, so liquefied α-amylase is used in actual gastrointestinal medicines. The blending ratio of saccharified α-amylase and saccharified α-amylase is preferably 4:1 to 1:1. As can be understood from the preferred embodiments described above, by effectively incorporating two or more starch digestive enzymes with different enzymatic actions into a gastrointestinal drug according to the present invention, faster efficacy and sufficient Digestion and absorption are achieved. Below, a method for separating and quantifying oligosaccharides by high performance liquid chromatography and various experimental results using this method will be described. Separation and quantification of oligosaccharides obtained by enzymatic decomposition of soluble starch Digestive enzymes were applied to soluble starch at 37°C, and after a certain period of time, the enzymes were deactivated by boiling. M. Somogyi; J.
Biol.Chem. 160 61 (1945)) to determine the titer. On the other hand, maltose and maltotriose present in this reaction solution were determined by high performance liquid chromatography. An outline of this quantitative method is shown in FIG. That is, as a column packing material.
After separating oligosaccharides using Nucleosil 10 NH ₂ , ethanolamine is pumped and heated to turn reducing sugars into phosphors (Toshio Kinoshita: Summary of the 65th Shimadzu High-speed Liquid Chromatography Conference, p. 3) (1980)) and quantified by post-label fluorescence spectroscopy. In this method, α-amylase is used as the digestive enzyme, and the chromatogram when it is reacted with soluble starch for 60 minutes and the chromatogram of a standard solution containing 5 nmol each of glucose, maltose, and maltotriose are shown in Figure 2. show. As is clear from FIG. 2, when α-amylase is applied to starch, the amounts of maltose and maltotriose produced are small, and the amount of oligosaccharides produced is extremely large. Classification of commercially available α-amylases according to mode of action Various types of commercially available α-amylases are allowed to act on starch, the amount of maltose and maltotriose produced is measured using the method described above, and based on the results α-amylase is
Amylases were classified into liquefaction type, saccharification type and intermediate type. The results are shown in Table 1.

[Table] Synergistic effect of liquefaction type α-amylase and saccharification type α-amylase Diasmen SS as liquefaction type α-amylase,
Biodiastase was blended in various proportions as saccharified α-amylase, and the amount of maltose and maltotriose produced was measured according to the above method,
The relationship between blending ratio and synergistic effect was investigated. The results are shown in Figure 3. In the figure, the rate of increase is the sum of the amount of sugar produced when liquefied α-amylase and saccharified α-amylase are used alone at each blending ratio (theoretical value).
100 from the percentage of sugar produced when blending with
Expressed by subtracting . Rate of increase = Amount of sugar produced during blending x 100 / Amount of sugar produced by liquefied α-amylase alone + Amount of sugar produced by saccharified α-amylase alone – 100 Combination of liquefied α-amylase and saccharified α-amylase The ratio was expressed not as a weight ratio but as a reducing sugar production ratio. The meanings of the symbols in the figure are as follows. -◆-: Rate of increase in maltose production -●-: Rate of increase in maltotriose production -△-: Rate of increase in maltose + maltotriose production As is clear from Figure 3, the difference between Diasmen SS and biodiastase A high synergistic effect can be obtained when the blending ratio is 4:1 to 1:3. Diasmen SS and biodiastase activity ratio: 7:
Synergistic effect when mixed with 2 Diasmene SS and biodiastase were mixed at an activity ratio of 7:2 and reacted in the same manner as above. Table 2 shows the results. Compared to the theoretical values, an approximately 20% increase in the amount of maltose and maltotriose produced was observed.

[Table] Example: Prepare granules whose daily dose is as prescribed below. Ingredient content (mg) Diasmen SS x 3 69 Biodiastase 2000 21 Prozyme 30 Sodium hydrogen carbonate 1200 Sanarmin 480 Precipitated sodium carbonate 480 Funnel extract 30 Cinnamon powder 60 Cucumber powder 15 Chiyoji powder 15 Wicker powder 3 Cinnamon oil 3 Cinnabar oil 3 Spruce oil 3 Carbonic acid Add starch and HPS (hydroxypropyl starch) as excipients to sodium hydrogen, sanarmin, and precipitated sodium carbonate, and make granules according to the conventional wet granulation method. On the other hand, Ziasmen SS
×3, Add excipients to Biodiastase 2000, Prozyme, Rohto extract, cinnamon powder, powdered cinnamon powder, powdered cinnamon powder, powdered fenugreek powder, cinnamon powder, cinnamon powder, and spruce oil, and similarly follow the conventional wet granulation method. Make granules, mix these two types of granules,
Separate the packaging.

[Brief explanation of the drawing]

Figure 1 is a systematic diagram of a sugar analysis system using high-performance liquid chromatography, Figure 2 is a chromatogram of the standard solution and α-amylase reaction solution, and Figure 3 is the synergy of liquefied α-amylase and saccharified α-amylase. This is a graph showing the effect. 1... Eluent, 2... Liquid pump, 3... Injector, 4... Column, 5... Fluorescent reaction liquid,
6... Reaction tank, 7... Stirrer, 8... Reaction coil, 9... Fluorescence detector, 10... Recorder, 11...
...glucose, 12...maltose, 13...maltotriose, 14...maltotetraose,
15... maltopentaose, 16... maltohexaose, 17... maltoheptaose, 18
...maltooctaose, a... rate of increase in the amount of maltose produced, b... rate of increase in the amount of maltotriose produced, c... rate of increase in the amount of maltose + maltotriose produced.

Claims (1)

[Claims] 1 Diasmene SS, a liquefied α-amylase
This is a gastrointestinal drug containing biodiastase, which is a saccharification type α-amylase, and the blending ratio is reducing sugar activity ratio Diasmen SS: Biodiastase =
Gastrointestinal drug with a ratio of 4:1 to 1:1.