Pepsin enhancers and their applications

Abstract

This invention provides a pepsin enhancer and its application, belonging to the field of digestive technology. The pepsin enhancer includes at least one of cytosine, uracil, guanine, thymine, and adenine. This invention also provides a pepsin enhancer oral solution, which is an aqueous solution of five bases with a concentration of 50-300 mg / L. This invention further provides chewable tablets, soft capsules, capsules, or tablets that promote digestion, containing five base powders. This invention provides a low-cost, non-toxic enhancer that is resistant to gastric acid degradation and improves pepsin activity. Oral solutions, chewable tablets, soft capsules, capsules, or tablets are prepared using this enhancer, which can significantly improve pepsin activity and aid digestion in humans and animals.

Description

Technical Field

[0001] This invention belongs to the field of digestive technology, specifically relating to pepsin enhancers and their applications. Background Technology

[0002] Functional dyspepsia is very common, and most people have experienced it. Its causes are mainly related to psychological factors, and it is particularly prevalent in elderly patients, significantly impacting their quality of life. Human digestion primarily relies on digestive enzymes secreted by the gastrointestinal tract. Western medicine generally supplements digestive enzymes to alleviate indigestion symptoms. Pepsin in gastric juice breaks down proteins in food into smaller peptides and peptones, and is the most important digestive enzyme in gastric juice, playing an indispensable role in the digestion of protein-rich foods. In addition, existing research has shown that pepsin plays an important role in peptic ulcers, gastric cancer, aspiration pneumonia, gastroesophageal reflux disease, and related reflux-related diseases such as reflux esophagitis, chronic rhinosinusitis, and otitis media. Traditional Chinese medicine uses strengthening the spleen and stomach as its main treatment methods. Commonly used traditional Chinese medicine preparations for treating indigestion, such as Baohe Wan, are related to their ability to significantly increase pepsin activity, which helps to relieve indigestion and treat food stagnation. Increasing the activity of pepsin can effectively improve symptoms such as abdominal discomfort, early satiety, nausea, vomiting, and bloating in patients, thereby achieving the therapeutic goal of improving patients' impaired digestive function.

[0003] This invention isolates an active strain of Bacillus subtilis X13 from persimmon enzyme. The fermentation supernatant of this strain, after high-temperature inactivation (100℃ for 20 min) and centrifugation at 10000 rpm for 5 min, significantly enhances pepsin activity, thereby improving pepsin's catalytic efficiency. Through separation, purification, and identification analysis of the fermentation supernatant, the inventors discovered that cytosine, uracil, and guanine in the fermentation supernatant all enhance pepsin activity. Further research showed that two other naturally occurring base molecules (thymine and adenine) also have pepsin-enhancing effects to varying degrees, with uracil exhibiting the most significant enhancing effect among the five bases. To date, there are no research reports, domestically or internationally, on the enhancing effect of bases on pepsin activity.

[0004] This invention has a wide range of applications. For example, by adding pepsin enhancers to food, the digestibility and utilization rate of protein by the human body can be improved, thereby enhancing the nutritional quality of food and improving human health. It can also be used to produce animal feed that improves breeding efficiency by assisting the activity of endogenous pepsin, increasing the digestibility and absorption rate of protein in animals, and thus promoting animal growth. Bases are endogenous molecules in humans and animals and are safe for living organisms. Therefore, this invention has promising development prospects and is of great significance for the development of digestive aids, health foods, and animal feed additives. Summary of the Invention

[0005] This invention first discovered a strain of Bacillus subtilis, isolated from a high-sugar persimmon enzyme stock solution prepared in the laboratory. The fermentation broth of this strain has the effect of enhancing pepsin activity. The inventors conducted in-depth research on this fermentation broth and discovered that the alkaline solution at a specific concentration has the effect of enhancing pepsin activity. This has been applied to pharmaceuticals and food, specifically including the following contents.

[0006] First, the present invention provides a pepsin enhancer, wherein the pepsin enhancer comprises at least one of cytosine, uracil, guanine, thymine, and adenine.

[0007] The synergistic dose of cytosine, uracil, and adenine is 0.036–0.054 mmol / L.

[0008] The synergistic dose of guanine and thymine is 0.022–0.045 mmol / L.

[0009] Second, the present invention provides a pepsin-enhancing oral solution, wherein the pepsin-enhancing oral solution is an aqueous solution of five bases, namely cytosine, uracil, guanine, thymine and adenine.

[0010] The concentration of the base aqueous solution is 50–300 mg / L.

[0011] The pepsin-enhancing oral solution also contains flavoring agents and preservatives.

[0012] The flavoring agent is sucrose, simple syrup, fructose syrup, orange peel syrup, citric acid syrup, cherry syrup, licorice syrup, cinnamon syrup, steviol glycosides, sorbitol, mannitol, citric acid, tartaric acid and vitamin C, and the preservative is potassium sorbate.

[0013] The flavoring agent has a content of 30-80 g / L, and the preservative has a content of 10-80 mg / L.

[0014] Finally, based on the above-mentioned mechanism of enhancing pepsin activity, the present invention also provides a chewable tablet, soft capsule, capsule or tablet that promotes digestion, wherein the chewable tablet, soft capsule, capsule or tablet contains five base powders, wherein the five bases are cytosine, uracil, guanine, thymine and adenine, wherein the content of cytosine, uracil and adenine is 1-50 mg, and the content of guanine and thymine is 1-30 mg.

[0015] The chewable tablets also contain compressible starch, microcrystalline cellulose, and skim milk powder.

[0016] Beneficial effects

[0017] Pepsin is crucial for the absorption and utilization of protein in the human body. For gastrointestinal dysfunction and various chronic gastrointestinal diseases caused by it, pepsin supplementation can have a positive impact on digestion and absorption, and as an adjunct therapy, compensating for deficiencies, declines, or loss of digestive function. It has advantages such as high specificity, low dosage requirements, and significant efficacy. Therefore, the development of pepsin-based products is of great significance to the nutrition and food industries. While the use of pepsin as a digestive aid and dietary supplement has been quite common in the West for decades, research on natural pepsin enhancers is still lacking both domestically and internationally.

[0018] The uracil studied in this invention, as a pepsin enhancer, can effectively increase the biochemical reaction rate of pepsin. When formulated into a mixed dietary supplement with pepsin, it can help improve gastrointestinal motility, facilitate digestion and nutrient absorption, and enhance the therapeutic effect of functional dyspepsia. Furthermore, uracil is a natural base and is harmless to humans and animals, while currently known pepsin enhancers such as Cu... 2+ Li + Even trace amounts of metal ions pose certain safety risks to the body. This is because excessive free Cu... 2+ It has damaging effects on cell membranes, proteins, and nucleic acids. It directly binds to esterified components in the TCA cycle, causing oligomerization of esterified proteins and loss of iron-sulfur cluster proteins, inducing protein toxicity stress, and ultimately leading to cell death. For Li + In terms of applications, the most serious cardiovascular adverse reaction of lithium salts is sinoatrial node dysfunction, which is one of the manifestations of cardiotoxicity. Therefore, when using lithium salts, extra caution should be taken regarding factors that may exacerbate lithium toxicity or lead to lithium accumulation. These are all factors that need to be considered when using Cu. 2+ Li + The potential safety risks of using trace amounts of metal ions as pepsin enhancers.

[0019] Furthermore, currently commercially available pepsin supplements are all derived from animal sources, while the pepsin enhancer studied in this invention can increase the activity of pepsin in humans or animals, undoubtedly greatly increasing its safety and effectiveness. The pepsin enhancer of this invention is a natural base, resistant to acid and high temperatures; the activity of pepsin increased by 1.48 times after treatment with the pepsin activator of this invention. The method for activating pepsin in this invention is simple, particularly suitable for preparing digestive health products and pharmaceuticals, with low cost and good market prospects. Attached Figure Description

[0020] Figure 1 The silica gel chromatography elution curve for Bacillus subtilis X13 pepsin enhancer.

[0021] Figure 2Elution curves of macroporous resin D-101 column chromatography for sample 6 of the distillate.

[0022] Figure 3 The image shows the spectral scans of the four distillate samples at wavelengths of 500-190 nm. Detailed Implementation

[0023] The present invention will be described in detail below with reference to examples. All methods and techniques are conventional.

[0024] In this invention, the following components were used: pepsin (30000 U / g), tyrosine (99% purity), hemoglobin, cytosine (99% purity); uracil (98% purity); thymine (99% purity); guanine (98% purity); adenine (98% purity), from Shanghai Yuanye Biotechnology Co., Ltd.; trichloroacetic acid (AR), from Shanghai Maclean Biochemical Technology Co., Ltd.; methanol (AR), petroleum ether (AR), ethyl acetate (AR), from Chengdu Kelong Chemical Co., Ltd.; acetone (AR), from Wuxi Zhanwang Chemical Reagent Co., Ltd.; silica gel (100-200 mesh), from Shanghai Haohong Biomedical Technology Co., Ltd.; and macroporous resin D-101, from Anhui Samsung Resin Co., Ltd.

[0025] The method for determining pepsin activity in this invention is as follows.

[0026] 1. Preparation methods of control solution and test solution.

[0027] (1) Acid solution: Measure 65 mL of 1 mol / L hydrochloric acid, add pure water to 1000 mL, mix well and set aside.

[0028] (2) Hemoglobin test solution: Weigh 1g of bovine hemoglobin, add acid solution to dissolve to 100mL, mix well and set aside.

[0029] (3) 5% trichloroacetic acid solution: Weigh 5g of trichloroacetic acid, add pure water to 100mL, mix well and set aside.

[0030] (4) Tyrosine control solution: Weigh 14.5 mg of tyrosine dried to constant weight at 105℃, add acid solution to 100 mL, mix well and set aside.

[0031] (5) Pepsin test solution: Weigh 0.01g of pepsin, add 12mL of acid solution, mix well to obtain a pepsin solution of 25U / mL, dilute the pepsin solution with acid solution to 0.25U / mL to obtain the pepsin test solution.

[0032] (6) Test sample solution: Take 2 mL of inactivated and cooled fermentation supernatant and 2 mL of pepsin test solution, mix well, and react in a water bath at 37℃ for 5 min to obtain the test sample solution experimental group. Take 2 mL of acid solution and 2 mL of pepsin test solution, mix well, and react in a water bath at 37℃ for 5 min to obtain the test sample solution control group.

[0033] 2. Method for determining pepsin activity.

[0034] Determination of pepsin in the experimental group: Take one test tube for the experimental group, add 5 mL of hemoglobin test solution, and incubate at 37℃ in a water bath. Add 1 mL of the test sample solution (either the experimental group or the control group), shake well, and incubate at 37℃ for 10 min. Immediately after incubation, add 5 mL of 5% trichloroacetic acid, shake well, and filter through three layers of filter paper. Collect the filtrate for later use. Take another test tube as the blank group, add 5 mL of hemoglobin, and incubate at 37℃ in a water bath for 10 min. Immediately after incubation, add 5 mL of 5% trichloroacetic acid, shake well, and add 1 mL of the test solution. Filter through three layers of filter paper. Collect the filtrate for later use. Measure the absorbance (A) of the experimental group filtrate at a wavelength of 275 nm using a spectrophotometer after zeroing the blank group.

[0035] Determination of the tyrosine control group: Take one test tube (make three replicates), add 1 mL of tyrosine control solution, incubate at 37℃ for 5 min, then add 5 mL of hemoglobin test solution preheated to 37℃, shake well, incubate at 37℃ for 10 min, immediately add 5 mL of 5% trichloroacetic acid solution, shake well, let stand at room temperature for 5 min, filter with slow qualitative filter paper, and collect the filtrate for later use. Take another test tube as the blank group, add 5 mL of hemoglobin test solution, incubate at 37℃ for 10 min, immediately add 5 mL of 5% trichloroacetic acid solution, shake well, and then add 1 mL of acid solution. Measure the absorbance (A') of the tyrosine control group filtrate at a wavelength of 275 nm using a spectrophotometer after zeroing the blank group.

[0036] The pepsin activity was calculated as shown in the formula below. It was incubated at 37°C for 10 min, and one unit of pepsin activity (U) was defined as the production of 1 μmol of tyrosine from the substrate per minute.

[0037]

[0038] In the formula, A: absorbance of the pepsin experimental group; A': absorbance of the tyrosine control group; W: sample weight, g; W S n: Amount of tyrosine in 1 mL of reference solution, in μg; n: Dilution factor of the test sample.

[0039] 3. Method for determining the activity of pepsin enhancers.

[0040] Pepsin enhancer activity (times / mL) = 2 × (psepsin activity in experimental group of test sample / pepsin activity in control group of test sample - 1)

[0041] Example 1

[0042] The applicant discovered that the high-sugar persimmon enzyme stock solution has a significant digestive aid function, and therefore studied the effective strains in it. The results showed that Bacillus subtilis X13 in one of the persimmon enzyme stock solutions has the effect of producing pepsin activators at high levels.

[0043] The extraction, separation, purification, and identification methods of the active ingredient of pepsin enhancer in the fermentation supernatant of Bacillus subtilis X13 are as follows.

[0044] (1) Methanol removal: The fermentation supernatant was concentrated 10 times by rotary evaporation (20 r / min, 40℃). An equal volume of methanol was added to the concentrate, and the mixture was shaken and placed in a 4℃ refrigerator. After standing for 1 hour, the mixture was centrifuged (10000 r / min, 10 min, 4℃). After centrifugation, the supernatant was collected and rotary evaporated (20 r / min, 40℃) to remove methanol. The concentrate was transferred to a separatory funnel for extraction.

[0045] (2) Petroleum ether extraction: Measure an equal volume of petroleum ether to the concentrate after methanol treatment and add it to a separatory funnel. Shake well and allow to stand for separation. After separation, place the lower concentrate and the upper organic phase separately in beakers. Repeat the extraction 5 times. Combine the separated organic phases and collect them in beakers. After extraction, rotary evaporate the organic phase (20 r / min, 40 °C) to remove the petroleum ether.

[0046] (3) Silica gel column chromatography: Weigh 30g of silica gel (100-200 mesh) and place it in a beaker. Add 50mL of a mixture of petroleum ether and ethyl acetate in a volume ratio of 9:1. Stir well and pour into the chromatography column. After filling, use a dropper to evenly drop the sample solution extracted by petroleum ether onto the surface of the silica gel bed. Elute with a petroleum ether and ethyl acetate eluent in a volume ratio of 9:1 at a elution rate of 5mL / min. The amount of eluent used is 600mL, and 50mL is one fraction. The obtained fraction 6 is evaporated to dryness by rotary evaporation (20r / min, 40℃) and then reconstituted with 5mL of pure water for later use.

[0047] (4) Macroporous resin column chromatography: Weigh 50g of macroporous resin D-101 and soak it in anhydrous ethanol for at least 24h. Wash it repeatedly with ultrapure water until it is free of ethanol. Weigh the pretreated macroporous resin and add 100mL of the sample solution extracted with petroleum ether (concentration 7.5 times that of the original solution). Stir well and place it in a shaking incubator (120r / min, 20℃, 2h) to allow the enzyme enhancer to be fully adsorbed into the macroporous resin. After adsorption is complete, load macroporous resin D-101 onto the column, filling the column to 1 / 2 of the column volume. Gently tap the column with a wash bottle during the process. After packing, drop fraction 6 evenly onto the surface of the macroporous resin bed. Elute with 500mL of acetone. No fraction collection is required during this process. Then elute with 200mL of acetone, 20mL for each fraction. Evaporate fraction 4 to dryness by rotary evaporation (20r / min, 40℃) and reconstitute with 2mL of pure water for later use.

[0048] Methods for identifying the active ingredients in pepsin enhancers

[0049] (1) Metabolite extraction: Transfer 50 μl of sample fraction 4 prepared in 1.2.3.2 into an EP tube, add 200 μl of extraction solution (methanol:acetonitrile = 1:1 (V / V)), the extraction solution contains isotope-labeled internal standard; vortex mix for 30 s, sonicate for 10 min (ice-water bath); let stand at -40℃ for 1 h; centrifuge the sample at 4℃, 12000 rpm for 15 min; take the supernatant into a sample vial for instrumental analysis.

[0050] (2) Analytical Separation: A Vanquish (Thermo Fisher Scientific) ultra-high performance liquid chromatograph was used to separate the target compounds using a Waters ACQUITY UPLC BEH Amide (2.1 mm × 50 mm, 1.7 μm) column. Phase A of the liquid chromatography was aqueous, containing 25 mmol / L ammonium acetate and 25 mmol / L ammonia, while Phase B was acetonitrile. Sample tray temperature: 4 °C, injection volume: 2 μl. The Orbitrap Exploris 120 mass spectrometer was able to acquire primary and secondary mass spectrometry data under the control of software (Xcalibur, version 4.4, Thermo). Detailed parameters are as follows: Sheath gas flow rate: 50 Arb, Aux gas flow rate: 15 Arb, Capillary temperature: 320℃, Full ms resolution: 60000, MS / MS resolution: 15000, Collision energy: SNCE 20 / 30 / 40, Spray voltage: 3.8kV (positive) or -3.4kV (negative).

[0051] (3) Data processing: The raw data was converted into mzXML format by ProteoWizard software, and then metabolite identification was performed using a collaboratively developed R package. The database used was BiotreeDB (V3.0) 7. Then, visualization analysis was performed using a self-developed R package.

[0052] The supernatant from the Bacillus subtilis X13 fermentation was purified by methanol removal, extracted with petroleum ether, and subjected to silica gel column chromatography. Fraction 6 was then collected. Figure 1 Then, fraction 6 was subjected to macroporous resin D101 column chromatography, and fraction 4 was collected. Figure 2 ), spectral scan of wavelengths from 500 to 190 nm ( Figure 3 It was found that fraction 4 of the distillate solution had absorption peaks at 194 nm and 265 nm in the ultraviolet spectrum. Substituting fraction 4 for liquid chromatography-mass spectrometry (LC-MS) analysis identified 208 level 1 organic compounds, meaning the metabolites in the sample matched the MS1, MS2, and RT values ​​of the standards. Based on the chemical characteristics of the pepsin enhancer produced by strain X13—namely, its weak polarity and absorption peaks at 194 nm and 265 nm in the ultraviolet spectrum—cytosine, uracil, and guanine were screened as potential target compounds for the following verification experiments.

[0053] Example 2

[0054] This embodiment verifies the synergistic effect of five bases on pepsin. The specific method is as follows: A certain amount of cytosine, uracil, thymine, guanine, and adenine were weighed respectively. Based on the concentration in the previous fermentation broth and the preliminary experiment, the five bases were prepared to concentrations of 0.022, 0.036, 0.045, and 0.054 mmol / L in sterilized and cooled YEPD liquid medium, respectively, to replace the fermentation supernatant in the preparation of the test solution. The rest of the method is the same.

[0055] Experiments showed that the solubility of the five bases in sterilized liquid culture medium was significantly higher than that in pure water. Furthermore, the sterilized liquid culture medium did not act as a pepsin enhancer and therefore did not interfere with the determination of the base synergistic effect. Thus, sterilized liquid culture medium was used to dissolve the bases in subsequent experiments. The experiments revealed that higher concentrations of cytosine, uracil, and guanine solutions did not exhibit pepsin synergistic effects, but upon dilution, they did show some pepsin synergistic effect. Uracil showed the most significant synergistic effect; as shown in Table 1, a 0.045 mmol / L uracil solution increased pepsin activity by 1.48 times. This experiment further measured the activity of the other two natural bases (thymine and adenine). These two natural bases also showed a similar trend, exhibiting pepsin synergistic activity only at lower concentrations, but their synergistic activity was lower than that of uracil.

[0056] Since pepsin activity is also affected by substrate concentration, meaning that when an synergist increases pepsin catalytic efficiency, but the substrate concentration is insufficient to support high-efficiency enzyme catalysis, the synergistic effect of the synergist may be affected, this experiment investigated the effect of increasing hemoglobin concentration on the synergistic effect of uracil. The results showed that when the hemoglobin concentration was increased by 0.5, 1, and 1.5 times, the synergistic activity of 0.045 mmol / L uracil was 1.12, 1.19, and 1.28 times that of the control group (with unchanged hemoglobin concentration), respectively. Furthermore, as shown in Table 1, at the three concentration gradients measured, the synergistic activity of the 0.054 mmol / L uracil solution was significantly lower than that of the 0.045 mmol / L solution. However, when the substrate concentration increased by 1.5 times, the synergistic activities of the 0.054 mmol / L and 0.066 mmol / L uracil solutions were 1.034 and 1.046 times that of the 0.045 mmol / L uracil solution, respectively. This indicates that the decrease in the synergistic activity of uracil at higher concentrations is due to insufficient substrate concentration. Therefore, when food is ingested, the substrate concentration of pepsin increases, and the synergistic effect of uracil also increases accordingly, without exhibiting the phenomenon of decreased synergistic effect at higher concentrations of uracil.

[0057] Table 1. Synergistic effects of five different concentrations of base solutions on pepsin

[0058]

[0059] Example 3

[0060] The inventors further determined whether there was a synergistic effect among the three highly synergistic bases (cytosine, uracil, and thymine). The results are shown in Table 2. The results indicate that, regardless of whether they were combined in pairs (1:1) or in groups of three (1:1:1), the synergistic activity of the combined solution was lower than the sum of the individual synergistic activities of each monomer. In particular, the synergistic activity of the uracil + thymine combination was reduced by 65% ​​compared to the sum of the individual activities of uracil and thymine. Therefore, to determine whether there is a synergistic effect among cytosine, uracil, and thymine, using uracil alone is the optimal choice.

[0061] Table 2. Synergistic effects of the combination solution of the three bases on pepsin.

[0062]

[0063] Example 4

[0064] This embodiment aims to investigate whether the duration of contact between uracil and pepsin in the stomach affects its synergistic effect when used as a pepsin enhancer in practical applications. The results are shown in Table 3. The synergistic activity of uracil is highest when in contact with pepsin for 5 minutes, but its synergistic activity is also high when the contact time is shorter. For example, uracil has a 1.05-fold synergistic activity when in contact with pepsin for only 1 minute, indicating that its onset time is relatively fast. Therefore, uracil is highly practical as a pepsin enhancer.

[0065] Table 3. Effect of contact time between uracil and pepsin on synergistic effect.

[0066]

[0067] Example 5

[0068] This embodiment provides several oral solutions that enhance pepsin activity, namely cytosine oral solution, uracil oral solution, guanine oral solution, thymine oral solution, and adenine oral solution.

[0069] The cytosine oral solution comprises cytosine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 250 mg of cytosine and 50 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 250 mg / L cytosine. Then add 50 mg of potassium sorbate to obtain the cytosine oral solution that enhances pepsin activity.

[0070] Uracil oral solution comprises uracil, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 255 mg of uracil and 50 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 255 mg / L uracil. Then add 50 mg of potassium sorbate to obtain a uracil oral solution that enhances pepsin activity.

[0071] The guanine oral solution consists of guanine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 270 mg of guanine and 50 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 270 mg / L guanine. Then add 50 mg of potassium sorbate to it to obtain a guanine oral solution that enhances pepsin activity.

[0072] The oral solution of thymine consists of thymine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 225 mg of thymine and 50 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 225 mg / L of thymine. Then add 50 mg of potassium sorbate to it to obtain an oral solution of thymine that enhances pepsin activity.

[0073] Adenine oral solution comprises adenine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 300 mg of adenine and 50 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 300 mg / L adenine. Then add 50 mg of potassium sorbate to obtain an adenine oral solution that enhances pepsin activity.

[0074] The cytosine oral solution comprises cytosine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 150 mg of cytosine and 30 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 150 mg / L cytosine. Then add 20 mg of potassium sorbate to obtain the cytosine oral solution that enhances pepsin activity.

[0075] Uracil oral solution comprises uracil, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 100 mg of uracil and 80 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 100 mg / L uracil. Then add 30 mg of potassium sorbate to obtain a uracil oral solution that enhances pepsin activity.

[0076] The guanine oral solution consists of guanine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 50 mg of guanine and 60 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 50 mg / L guanine. Then add 80 mg of potassium sorbate to it to obtain a guanine oral solution that enhances pepsin activity.

[0077] The oral solution of thymine consists of thymine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 120 mg of thymine and 60 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 120 mg / L thymine. Then add 40 mg of potassium sorbate to it to obtain an oral solution of thymine that enhances pepsin activity.

[0078] Adenine oral solution includes adenine, sucrose, and potassium sorbate. The specific preparation method is as follows: accurately weigh 180 mg of adenine and 30 g of sucrose and dissolve them in 1 L of deionized water to prepare a sucrose solution with a final concentration of 180 mg / L adenine. Then add 10 mg of potassium sorbate to obtain an adenine oral solution that enhances pepsin activity.

[0079] Preliminary animal experiments showed that the activity of pepsin in mice increased by 0.5 to 1.2 times after oral administration of 1 mL of the alkaline oral solution prepared by the above method.

[0080] Sucrose is added as a flavoring agent. Sucrose can be replaced with simple syrup, fructose syrup, orange peel syrup, citron syrup, cherry syrup, licorice syrup, and cinnamon syrup. Stevioside, sorbitol, and mannitol can also be used as flavoring agents. To improve the taste, citric acid, tartaric acid, and vitamin C can be added at a concentration of 1–10 g / L.

[0081] Example 6

[0082] A method for preparing a digestive alkaline chewable tablet, the main components of which include compressible starch, microcrystalline cellulose, skim milk powder and five kinds of alkaline powder, is prepared by compressing with a tableting machine. In this embodiment, the formulation of a compressed tablet is as follows.

[0083] Table 4. Formulation of Alkaline Chewable Tablets

[0084]

[0085] Similarly, since the above embodiments have demonstrated that the core effective component of the present invention is a single base, namely cytosine, uracil, guanine, thymine, and adenine, other forms of packaging, such as soft capsules, capsules containing 1-5 mg of cytosine, uracil, or adenine, or containing 1-3 mg of guanine or thymine, can also achieve the effect of increasing pepsin activity.

[0086] Example 7

[0087] A drug to promote animal digestion is administered in the form of soft capsules or capsules. Each soft capsule or capsule contains 1–50 mg of cytosine, uracil, or adenine, or 1–30 mg of guanine or thymine. The dosage is determined based on the animal's gastric acid levels and the average water content of a meal. Adding this drug can increase the activity of pepsin in the animal, thereby aiding digestion. Preliminary animal experiments have shown that pepsin activity in mice can be increased by 0.5–1.3 times.

Claims

1. The application of a five-base solution, wherein the five-base solution comprises one of cytosine, uracil, guanine, thymine, and adenine, characterized in that: The base solution is used to prepare a pepsin enhancer, wherein the enhancing dose of cytosine, uracil, and adenine is 0.036~0.054 mmol / L; and the enhancing dose of guanine and thymine is 0.022~0.045 mmol / L.

2. The application of a five-base solution, wherein the five-base solution comprises one of cytosine, uracil, guanine, thymine, and adenine, characterized in that: The base solution is used to prepare a pepsin-enhancing oral solution, and the concentration of the base aqueous solution is 50~300 mg / L.

3. The application of the five base solutions according to claim 2, characterized in that: The pepsin-enhancing oral solution also contains flavoring agents and preservatives.

4. The application of the five base solutions according to claim 3, characterized in that: The flavoring agents are sucrose, simple syrup, fructose syrup, orange peel syrup, citric acid syrup, cherry syrup, licorice syrup, cinnamon syrup, steviol glycosides, sorbitol, mannitol, citric acid, tartaric acid, and vitamin C, and the preservative is potassium sorbate.

5. The application of the five basic solutions according to claim 3, characterized in that: The flavoring agent content is 30~80g / L, and the preservative content is 10~80mg / L.

6. The application of a five-base powder, wherein the five bases include one of cytosine, uracil, guanine, thymine, and adenine, characterized in that: The base powder is used to prepare pepsin-enhancing chewable tablets. The formula of the chewable tablets is: 80-150 parts compressible starch, 100-160 parts microcrystalline cellulose, 70-140 parts skim milk powder, and 1-5 parts base powder.

7. The application of a five-base powder, wherein the five bases include one of cytosine, uracil, guanine, thymine, and adenine, characterized in that, The base powder is used to prepare pepsin-enhancing capsules or tablets, wherein the contents of cytosine, uracil, and adenine in the pepsin-enhancing capsules or tablets are 1-50 mg, and the contents of guanine and thymine are 1-30 mg.

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