Lamb stomach extract vitamin B12 preparation and its use in the treatment of gastric disorders

Abstract

Lamb stomach extract Vitamin B 12 A pharmaceutical composition comprising a lamb stomach extract and vitamin B 12 The lamb stomach extract has a chymosin titer of more than 40 U / g and a pepsin titer of more than 15 U / g, and is a mixture of lamb stomach extract and vitamin B 12 and their use in treating moderate to severe chronic atrophic gastritis, reversing intestinal metaplasia and preventing gastric cancer. 12 showed synergistic effects or interactions at certain ratios.

Description

[Technical field]

[0001] The present invention belongs to the field of medicine and relates to the efficacy of lamb stomach extract vitamin B in treating moderate to severe chronic atrophic gastritis, reversing intestinal metaplasia and preventing gastric cancer. 12 Use of the pharmaceutical composition. [Background technology]

[0002] Chronic gastritis is the most common type of digestive disease, and chronic atrophic gastritis is a type of chronic gastritis, accounting for about one-third of chronic gastritis cases. Clinically, some patients with chronic atrophic gastritis may not have obvious symptoms. However, most patients may complain of symptoms such as epigastric burning pain, bloating, dull pain or fullness, loss of appetite, nausea, and borborygmus, especially after eating. In severe cases, emaciation, anemia, brittle nails, and inflammation of the tongue and atrophy of the lingual papillae may also be present.

[0003] Chronic atrophic gastritis (CAG) is characterized by atrophy of gastric glands, mucosal thinning, thickening of the muscularis mucosae, and associated intestinal metaplasia and atypical hyperplasia. Its diagnosis depends primarily on gastroscopic findings and pathological findings of gastric mucosal biopsies.

[0004] The disease progression cycle from normal gastric mucosa to gastric cancer is: normal gastric mucosa → nonatrophic gastritis → atrophic gastritis → intestinal metaplasia → intraepithelial neoplasia → gastric cancer. Due to the above pathological progression, gastric diseases are divided into nonatrophic gastritis, atrophic gastritis, intestinal metaplasia, intraepithelial neoplasia, and gastric cancer. There are no effective drugs to reverse atrophic gastritis, intestinal metaplasia, or intraepithelial neoplasia. Summary of the Invention

[0005] In order to solve the above problems, the present invention aims to provide a method for the preparation of lamb stomach extract containing vitamin B 12 The present invention provides a formulation.

[0006] Another object of the present invention is to provide the lamb stomach extract with vitamin B 12 Use of the pharmaceutical composition is provided.

[0007] In order to achieve the above object, the present invention provides a lamb stomach extract containing vitamin B 12 A pharmaceutical composition comprising a lamb stomach extract and vitamin B 12 The lamb stomach extract has a chymosin titer of more than 40 U / g and a pepsin titer of more than 15 U / g, and is a mixture of lamb stomach extract and vitamin B 12 The weight ratio of the formulation composition to the active ingredient is 100,000:0.5-5.

[0008] Preferably, the lamb stomach extract and vitamin B 12 The weight ratio is 100000:1 to 4.5.

[0009] More preferably, the lamb stomach extract and vitamin B 12 is 100000:1, 100000:2, 100000:3, 100000:4 or 100000:4.5 by weight.

[0010] Preferably, the chymosin titer is 45 U / g or more and the pepsin titer is 20 U / g or more; alternatively, the chymosin titer is 55 U / g or more and the pepsin titer is 20 U / g or more.

[0011] Preferably, the protein of the lamb stomach extract has a relative molecular weight of 10,000 to 45,000.

[0012] Preferably, the ratio of the chymosin titer to the pepsin titer is from 2.5 to 5.1.

[0013] As mentioned above, it further comprises a pharma- ceutically acceptable additive, preferably one or more of an excipient, a binder, a diluent, a flow aid, a flavor enhancer, an antioxidant, and a bacteriostatic agent.

[0014] As described above, the formulation may be in the form of capsules, tablets, suspensions or granules.

[0015] The present invention also relates to a method for the preparation of a medicament for reversing gastric mucosal atrophy comprising administering to the patient a dose of vitamin B12 from a lamb stomach extract. 12 Uses of the pharmaceutical compositions are provided.

[0016] The present invention also relates to the use of lamb stomach extract vitamin B in the manufacture of a medicament for treating moderate or severe chronic atrophic gastritis. 12 Uses of the pharmaceutical compositions are provided.

[0017] The present invention also relates to a method for treating intestinal metaplasia using lamb stomach extract vitamin B 12 Uses of the pharmaceutical compositions are provided.

[0018] As described above, the intestinal metaplasia is chronic atrophic gastritis associated with intestinal metaplasia.

[0019] The present invention also relates to a method for the preparation of a medicament for treating gastric intraepithelial neoplasia using lamb stomach extract vitamin B 12 Uses of the pharmaceutical compositions are provided.

[0020] The present invention also relates to a method for the preparation of a medicament for preventing gastric cancer, comprising administering to the patient a dose of vitamin B12 from a lamb stomach extract. 12 Uses of the pharmaceutical compositions are provided.

[0021] As stated above, the treatment is an effective treatment.

[0022] As stated above, the treatment is a reversal treatment.

[0023] As described above, the subjects for administration are Helicobacter pylori-negative patients.

[0024] As mentioned above, antioxidant pathways provide for the treatment, reversal and prevention of the above.

[0025] As mentioned above, the treatment, reversal or prevention of the above can be achieved by modulating the intestinal flora dysbiosis.

[0026] As stated above, the formulation is administered to patients at 330 U, three times per day.

[0027] Lamb stomach extract and vitamin B provided by the present invention 12 The proportional dosage of the formulation has a direct impact on the therapeutic effect. Specifically, the specific ratio of lamb stomach extract and vitamin B 12 The synergistic effect was observed when lamb stomach extract or vitamin B 12 Lamb stomach extract and vitamin B compared with 12 The weight ratios of vitamin B were observed in the range of 100,000:4 to 100,000:1. For a weight ratio of 100,000:4, 12 The lamb stomach extract combined with the lower weight ratio of 100,000:1 was favorable for suppressing the remaining fundic mucosal congestion. Furthermore, vitamin B 12 When the weight ratio was decreased to 100000:0.2, no synergistic effect of the formulation was obtained.

[0028] The lamb stomach extract of the present invention contains vitamin B 12 The preparation comprises a lamb stomach extract, said preparation having a chymosin titer of 45 U / g or more and a pepsin titer of 20 U / g or more; or a lamb stomach extract having a chymosin titer of 55 U / g or more and a pepsin titer of 20 U / g or more.

[0029] The beneficial effects of the present invention are as follows: The present invention has demonstrated that lamb stomach extract vitamin B can effectively reverse gastric mucosal atrophy, intestinal metaplasia and intraepithelial neoplasia, significantly reduce the OLGA and OLGIM stages of severe CAG patients, and effectively reduce the risk of gastric cancer. 12 Pharmaceutical compositions are provided. [Brief description of the drawings]

[0030] [Figure 1] FIG. 1 is a graph showing the results of comparing the gastric juice pH of rats in a negative control group, a treatment group, and a non-treatment group, each of which was administered a lamb stomach extract vitamin B12 preparation provided by the present invention. [Diagram 2]FIG. 2 (AF) is a graph showing the pathological changes in the gastric mucosa of rats in the negative control group, the treatment group, and the non-treatment group, caused by the lamb stomach extract vitamin B12 preparation provided by the present invention. [Diagram 3] FIG. 3 shows the effective rate of chronic inflammation by site in patients with chronic atrophic gastritis, using the lamb stomach extract vitamin B12 preparation provided by the present invention. [Figure 4] FIG. 4 shows the incidence of reversal of chronic inflammation by site / effectiveness rate of inflammatory activity by site in patients with chronic atrophic gastritis by the lamb stomach extract vitamin B12 preparation provided by the present invention. [Diagram 5] FIG. 5 shows the effective rate of atrophy by site in patients with chronic atrophic gastritis treated with the lamb stomach extract vitamin B12 preparation of the present invention. [Figure 6] FIG. 6 shows the incidence of reversal of atrophy by site in patients with chronic atrophic gastritis by the lamb stomach extract vitamin B12 preparation provided by the present invention. [Figure 7] FIG. 7 shows the effectiveness of the lamb stomach extract vitamin B12 preparation provided by the present invention in intestinal epithelial chemotaxis by site in patients with chronic atrophic gastritis. [Figure 8] FIG. 8 shows the incidence of reversal of intestinal metaplasia by site in patients with chronic atrophic gastritis using the lamb stomach extract vitamin B12 preparation provided by the present invention. [Figure 9] FIG. 9 shows a comprehensive evaluation of the efficacy of the lamb stomach extract vitamin B12 preparation provided by the present invention on gastric chronic inflammation, atrophy and intestinal metaplasia in patients with chronic atrophic gastritis. [Figure 10] FIG. 10 shows a comprehensive evaluation of the incidence of reversal of gastric chronic inflammation, atrophy and intestinal metaplasia in patients with chronic atrophic gastritis by the lamb stomach extract vitamin B12 preparation provided by the present invention. [Figure 11] Figure 11 shows the gastroscopic findings of the patient on May 13, 2019, with A being the pyloric antrum and B being the body of the stomach. [Figure 12] Figure 12 shows the histopathological findings of a biopsy taken from a patient on May 13, 2019, stained with hematoxylin and eosin at low magnification; A: pyloric antrum, B: gastric body. [Figure 13] Figure 13 shows the gastroscopic findings of the patient on December 11, 2019, with A being the pyloric antrum and B being the body of the stomach. [Figure 14] Figure 14 shows the histopathological findings of a biopsy from a patient on December 11, 2019, stained with hematoxylin and eosin at low magnification; A: pyloric antrum, B: gastric body. [Figure 15] Figure 15 shows the gastroscopic findings of the patient on March 30, 2021, with A being the pyloric antrum and B being the body of the stomach. [Figure 16] Figure 16 shows the histopathological findings of a patient's biopsy on March 30, 2021, stained with hematoxylin and eosin at low magnification; A: pyloric antrum, B: gastric body. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] In order to make the advantages and features of the present invention more easily understood by those skilled in the art and to define the protection scope of the present invention more clearly and explicitly, the following embodiments of the present invention are described in detail and completely. It should be understood that the specific embodiments described herein are for the purpose of illustrating the present invention and are not intended to limit the present invention.

[0032] Lamb stomach extract Vitamin B 12 The oral formulation or granules contain highly active organisms extracted from the rumen of Xinjiang Tianshan sheep (containing various physiologically active ingredients such as chymosin, pepsin, mucin, and bifidogenic factors) and vitamin B 12 The main component is. EXAMPLES

[0033] Example 1 Extraction and isolation / purification of ruminal enzymes from lamb stomach extract Lamb stomach extract was defatted with petroleum ether and extracted with 0.9% sodium chloride aqueous solution at low temperature under ultrasonic assistance. The extraction solution was 0.5 mol / L hydrochloric acid at pH 4.6. The extract was precipitated with 30%, 50%, and 90% saturated ammonium sulfate successively, and the final 90% saturated ammonium sulfate precipitate was the total enzyme active site. The precipitate was dissolved in deionized water, and the dialysis bag was desalted, and then separated and purified with diethylaminoethyl (DEAE) cellulose 52 anion exchange resin. The sample was eluted with a gradient of equilibration solution (0.05 mol / L phosphate buffer at pH 5.7) and equilibration solution containing 0.1, 0.2, 0.3, 0.4, and 0.5 mol / L NaCl solutions, and the eluate was detected by ultraviolet (UV) detection at 280 nm. The effluent was collected at a speed of 10 min / tube, and the chymosin titer was measured. The 0.4 and 0.5 mol / L NaCl aqueous solution elution sites (designated F6 and F7) had chymosin activity, and were further purified by high pressure liquid chromatography (HPLC) using a gel chromatography column with 0.9% NaCl aqueous solution as the eluent at a flow rate of 0.4 mL / min. Protein absorption was detected at two wavelengths, 212 nm and 280 nm, and samples were collected. Chymosin titers of the samples were measured by the Arima method, and pepsin titers were measured by the Mazorra Manzano method. The samples were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using a 15% separation gel. F6-2 was analyzed by LC MS / MS, and F6-2 was enzymatically cleaved into small peptides by trypsin, and the peptide fragments were analyzed by LC MS / MS, and the analytical data were identified using the similarity search tool of the SIB ExPASy bioinformatics database.

[0034] The enzyme activity and yield at each stage of isolation and purification of lamb stomach extract are shown in Table 1. The chymosin titer of lamb stomach extract was (691.2±5.16) U / g, and after one-step purification by saturated ammonium sulfate precipitation, the chymosin titer was (18341.61±4.1) U / g, which increased 26.5-fold. As a result of separation and purification by total enzyme active site chromatography on a DEAE-cellulose 52 anion exchange column, among the seven elution sites, F6 and F7 had chymosin titer, and F7 also had pepsin activity. The isolated and purified chymosins F6-2 and F7-2 had high chymosin activity and low casein hydrolysis activity, while pepsin had bovine serum albumin hydrolysis activity, and F7-1 had this measurement characteristic. As a result of HPLC separation and purification profile, F6-2 and F7-2 were chymosin, and F7-1 was pepsin, and the enzyme titers were (27557.1 ± 4.8) U / g, (17532.6 ± 5.1) U / g and (17728.15 ± 5.3) U / g, respectively. The relative molecular weights of F6-2, F7-2 and F7-1 were all in the range of 35000 to 40000. The results of LC-MS / MS (nano LC-QE) analysis of chymosin and database comparison are shown in Table 2. The relative molecular weight of chymosin F6-2 was 40392.88 and the isoelectric point was 4.58.

[0035] [Table 1]

[0036] [Table 2]

[0037] Example 2 Analysis of antioxidant activity of lamb stomach extract The lamb stomach extract was acid hydrolyzed, derivatized with DNFB, and analyzed for amino acid composition using HPLC Diamonsil AAA amino acid analysis column. To completely dissolve the drug substance, it was extracted sequentially with water, 0.1 mol / L phosphate buffer at pH 7.0, and 0.01 mol / L ammonium bicarbonate aqueous solution. The extract was desalted, lyophilized, and pre-stored at -20°C. The three extracts were assayed for in vitro antioxidant activity of 1,1-diphenyl-2-trinitrophenylhydrazine [DPPH] and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate) diammonium salt [ABTS] free radicals using the method reported in

[13] . A 15% separation gel for SDS-PAGE was used to analyze the relative molecular weight range of proteins in the three extracts.

[0038] The amino acid composition analysis results of the lamb stomach extract are shown in Table 3. Hydrophilic amino acids directly involved in the antioxidant reaction process accounted for 33.04% of the total amino acid content. The in vitro antioxidant activities of the three extracts extracted from the drug substance aqueous solution, phosphate buffer solution, and ammonium bicarbonate solution were (37.08±0.45)%, (23.20±0.78)%, and (62.80±0.74), respectively, and the DPPH scavenging activities were (57.87±0.55)%, (5.03±0.25)%, and (26.67±0.21), respectively. The comparison of antioxidant activities showed that the aqueous extract and ammonium bicarbonate extract had higher ABTS scavenging activity and DPPH scavenging activity than the phosphate buffer extract, and thus had higher antioxidant activity. The results of SDS-PAGE of the three extracts showed that the relative molecular weight of the protein in the drug substance was 10,000-40,000.

[0039] [Table 3]

[0040] Analysis of crude glycoprotein extracts from lamb stomach and their probiotic activity From the results in Table 4, the crude glycoprotein extract showed growth-promoting effects on Bifidobacterium adolescentum, Lactobacillus terreus Bulgaria subspecies, and Enterococcus faecalis, and the difference in the growth-promoting effects on Lactobacillus terreus Bulgaria subspecies and Enterococcus faecalis was statistically significant (P<0.05 for both). The glycoproteins had overlapping protein and sugar elution peaks in the chromatogram elution curve, and GP was a glycoprotein purified with Sephadex G-50, while GP-1 was a glycoprotein separated and purified with DEAE-cellulose 52. The total sugar content of GP-1 was 70.24%, the protein content was 16.39%, the polysaccharide chain consisted of two monosaccharides, lactose and glucose, and the amino acid composition was as shown in Table 5.

[0041] [Table 4]

[0042] [Table 5]

[0043] When the antioxidant activity of three types of extracts of the drug substance was examined, antioxidant activity was found in all extracts, with the water extract and ammonium bicarbonate extract showing higher antioxidant activity, suggesting that they are the main components that play a pharmacological role in the treatment and prevention of gastric ulcers, chronic atrophic gastritis, and gastric cancer.

[0044] Example 3 Lamb stomach extract Vitamin B 12 Subacute toxicity test of chymosin and pepsin activity using formulations. I. Materials and Methods 1. Animals: Thirty healthy rats weighing 115-195g (collected from the animal house and fed for one week before use in the experiment) were divided into three groups of 10 rats each. 2. Sample: Lamb stomach extract Vitamin B 12 formulation 3. Methods: Three groups of animals were divided into a control group, a low-dose maximum group, and a high-dose maximum group. The control group was given 1.5 ml of saline orally daily for 30 days. The low dose group was administered 500 mg / kg body weight daily for 30 days, and samples were prepared as 1.5 ml suspensions in saline. The high dose group was administered 1500 mg / kg body weight daily for 30 days, and samples were prepared as 1.5 ml suspensions in saline. 4. Observation items: Before and 30 days after administration of this drug, the animals' serum glutamic pyruvic transaminase (SGPT) activity and serum non-protein nitrogen content were measured, and 30 days after administration of this drug, pathological sections of the animals' organs such as the liver, spleen, heart, kidney, small intestine, and stomach were observed to observe the effects of this drug on the animals' liver function, renal function, and pathological changes in the organs.

[0045] Serum glutamic pyruvic transaminase (SGPT) measurement method Serum glutamic-pyruvic transaminase is highest in hepatocytes, and when certain drugs cause liver damage or when a patient is in the acute phase of viral hepatitis, liver cells are damaged and the enzyme overflows into the blood, causing a significant increase in serum glutamic-pyruvic transaminase activity. Therefore, changes in serum glutamic-pyruvic transaminase activity can be used as an important indicator of subacute drug poisoning and liver disease.

[0046] 2.Measurement method (1) Reagents 1. Pyruvic acid standard solution: 23.5 mg of sodium pyruvate was dissolved in 0.1 M pH 7.4 phosphate buffer and the solution was adjusted to a final volume of 100 ml. 2. Substrate solution: Dissolve 87.6 mg of ketoglutaric acid and 5.34 g of dl-alanine in 90 ml of 0.1 M, pH 7.4 phosphate buffer, adjust the pH to 7.4 with 20% NaOH solution, then dilute with 300 ml of the above phosphate buffer, store in the refrigerator, and use for one week. 3. 2,4-Dinitrophenylhydrazine solution: 19.8 mg of 2,4-dinitrophenylhydrazine was dissolved in 8.0 ml of concentrated hydrochloric acid, transferred to a 100 ml measuring flask, and made up to the mark with distilled water. 4.0.4N NaOH solution 5. 0.1M, PH 7.4 Phosphate Buffer: 2.69g KH2PO4 and 13.97g K2HPO4 were dissolved in water, transferred to a 1000ml volumetric flask and diluted with distilled water.

[0047] (2) Preparation of calibration curve: Six dry, clean test tubes were numbered 0, 1, 2, 3, 4, and 5, and 0, 0.05, 0.10, 0.15, 0.20, and 0.25 ml of pyruvate standard solution was added, and 0.25, 0.20, 0.15, 0.10, 0.05, and 0 ml of 0.1M, pH 7.4 phosphate buffer was added. Then, 0.25 ml of SGPT substrate solution and 0.5 ml of 2,4-dinitrophenylhydrazine solution were added, mixed, and kept in a 37°C water bath for 20 minutes. 5.0 ml of 0.4N NaOH rye was added to each tube, mixed, and left at room temperature for 10 minutes. The zero point was adjusted with distilled water, and the absorbance of each tube was measured at a wavelength of 520 nm using a 750 spectrophotometer equipped with a 200u microcell. The absorbance was measured at 520 nm and a calibration curve was constructed with the difference in optical density between each tube and the "0" tube as the ordinate and the corresponding micrograms of pyruvate as the abscissa.

[0048] (III) SGPT titer assay Preparation of animal serum: 0.5-1.0 ml of blood was taken from rats by cutting their tails, allowed to clot, and centrifuged at 3000 rpm for 10 minutes, and serum was collected and set aside.

[0049] Several dry and clean test tubes were taken, one of which was a control tube. For the remaining measurement tubes, stomach was microinjected and 0.02 ml of serum was added to each tube, and 0.10 ml of SGPT substrate solution was added to the measurement tube and mixed. The tubes were incubated in a 37°C water bath for 30 minutes, and 0.10 ml of 2,4-dinitrophenylhydrazine solution was added to each tube, while 0.10 ml of SGPT substrate solution was added to the control tube, mixed, and incubated in a 37°C water bath for 20 minutes. The zero point was adjusted with distilled water, and the light absorption was measured at a wavelength of 520 nm. The number of micrograms of pyruvate was determined from the calibration curve, and the SGPT activity was calculated according to the following formula:

[0050]

number

[0051] 3. Measurement of serum non-protein nitrogen content Serum non-protein nitrogen is a general term for nitrogen contained in substances such as urea, uric acid, amino acids, ammonia, and bilirubin, and mainly refers to urea nitrogen. A significant increase in serum non-protein nitrogen indicates kidney damage. Therefore, the measurement of serum non-protein nitrogen content can be used to estimate the effect of subacute drug poisoning on the kidney and the degree of kidney damage.

[0052] Measurement method (1) Reagents 1.10% sodium tungstate solution 2.2 / 3N NaOH solution 3.1:1 Sulfuric acid 4. Nessler's Reagent: 22.5g of iodine, 30g of KI, and 20ml of water were dissolved in a 500ml flask, 30g of pure water was added, and the mixture was shaken well. The mixture was then rinsed with tap water and cooled intermittently, and shaken until the yellow color of the supernatant disappeared. The supernatant was poured into a 200ml measuring flask, diluted to the mark, mixed, and 975ml of 2.5N NaOH aqueous solution was added and mixed. The mixture was stored in a brown bottle to clarify it and serve as a spare. 5. Standard ammonium sulfate solution (0.06 mg nitrogen per ml): Pure ammonium sulfate was placed in a desiccator for several days, then 0.2834 g was taken into a 10 ml measuring flask, diluted to the mark and mixed well (at this point this stock solution was taken and diluted 2-fold to give a working solution containing 0.03 mg nitrogen per ml).

[0053] (3) Measurement 1. Preparation of protein-free filtrate: Add 0.7 ml of water, 0.1 ml of serum, and 0.1 ml of 10% sodium tungstate to a 5 ml dry, clean small centrifuge tube, mix well, then add 0.1 ml of 2 / 3N H2SO4 solution, mix well, leave for 10 minutes, then centrifuge at 3000 RPM for 10 minutes, and reserve the supernatant. 2. Prepare three hard test tubes: one for measurement, one for standard, and one for blank. 0.5 ml of protein-free filtrate and 0.1 ml of 1:1 sulfuric acid were added to the measurement tube, mixed thoroughly, and then digested under an alcohol lamp until colorless and cooled. 0.5 ml of standard ammonium sulfate solution, 0.1 ml of 1:1 sulfuric acid, and 4.4 ml of distilled water were added to the standard tube. 5.0 ml of distilled water was added to the blank tube. 1.5 ml of Nessler's reagent was added to the above three tubes and mixed thoroughly. The blank tube was used to adjust the absorbance to zero, and the absorbance at 420 nm was measured. The non-protein nitrogen content per 100 ml of serum was calculated according to the following formula.

[0054]

number

[0055] In a normal individual, 100 ml of blood contains 21 to 42.5 mg of nonprotein nitrogen.

[0056] 4. Discussion of the results [Table 6]

[0057] [Table 7]

[0058] [Table 8]

[0059] [Table 9]

[0060] As can be seen from Tables 6 to 9, compared with normal rats, before administration, the SGPT titer of 2 out of 30 rats was 40 units or more, and the NPN content of 10 rats was 42.5 mg% or more, but after administration, the SGPT titer of all rats was 40 units or less. Regarding the NPN content, after administration, the serum NPN content of all rats was within the normal range, except for 2 rats in the control group, which had an NPN content of 42.5 mg% or more. From this, it can be seen that the lamb stomach extract vitamin B 12 The formulation was shown to cause no damage to the liver and kidneys of rats.

[0061] 5. Observation of pathological sections Thirty days after administration, the rats were sacrificed and parts of the liver, lungs, heart, spleen, kidneys, stomach, and small intestine were immediately removed, fixed in 10% formaldehyde solution, and sectioned. No significant differences were observed between the control and treatment groups.

[0062] Example 4 Effect of vitamin B from lamb stomach extract on precancerous lesions in rat gastric mucosa 12 Therapeutic effect of the formulation Research method 1: Testing agent: The lamb stomach extract under test had a chymosin titer of 45 U / g, pepsin titer of 20 U / g, and a relative molecular weight of protein of 10,000-40,000. 12 is a USP Vitamin B 12 It was.

[0063] Control and treatment groups The animal distribution is as follows: - The control group (A) was given only po water under the same conditions as the other groups. - Treatment group (B) received lamb stomach extract: vitamin B 12 A combination of 100,000:1 by weight was used. - Treatment group (C) received vitamin B 12 Only lamb stomach extracts free of . - Treatment group (D) received lamb stomach extract: vitamin B 12 A combination of 100,000:0.2 by weight was used. - Treatment group (E) received lamb stomach extract: vitamin B 12 A combination of 100,000:4 by weight was used.

[0064] The treatments for each group were performed under the same conditions, with vitamin B 12 The test combination was formulated in an aqueous suspension and administered orally at a dose of 400 mg / kg, except for the group treated with vitamin B alone. 12 was administered at (4ug / kg).

[0065] Analysis of covariance (ANCOVA) was applied to the data for each day to perform statistical comparisons of body weight changes in the different experimental groups.

[0066] The incidence of peptic mucosal ulcers and inflammation in different experimental groups is shown in Table 10. [Table 10]

[0067] As can be seen from Table 6, treatment group B showed a significant improving effect on the appearance of inflammation of the gastrointestinal mucosa. This effect was observed in all aspects of the gastrointestinal tract observed at autopsy. Groups E and B showed the same clinical effect, and the protective effect was also similar. The number of congestions in the residual mucosal base was the lowest in group B compared to group E.

[0068] By comparison, vitamin B 12 Group C, which received only lamb stomach extract without vitamin B12, was similar to control group A. 12The results were similar to those of group F, which received only rituximab. The severity of gastrointestinal disorders was similar in groups B and E. Group D showed only a slight improvement over the results of control group A.

[0069] As can be seen from Table 6, lamb stomach extract and vitamin B 12 In terms of efficacy, both treatment groups C (vitamin B 12 Treatment group B (lamb stomach extract) had no protective effect against phenylbutazone-induced ulcers, whereas treatment group C (lamb stomach extract) was highly protective (P<0.00001).

[0070] Vitamin B 12 No protective effect was observed in the treatment group (D), which had the lowest content. On the other hand, the protective effect provided by treatment group (E) was compared with treatment group (B), and treatment group (B) had a lower number of residual basal mucosal congestions observed than treatment group (E). These results indicate that lamb stomach extract and vitamin B 12 When the weight ratio of is in the range of 100,000:4 to 100,000:1, lamb stomach extract or vitamin B 12 A synergistic effect was observed when compared with the use of either vitamin B alone. The combination of 100,000:1 was more favorable for suppressing congestion of the remaining basal mucosa than the lower ratio of 100,000:4. 12 When the weight ratio was further reduced to 100,000:0.2, no synergistic effect was observed when the two were combined. 12 Formulations were prepared according to the optimal combination B for subsequent studies.

[0071] Research method 2: 1. Experimental animals: 42 specific pathogen-free grade healthy Wistar male rats (weight 140-180g, 4-6 weeks old) were selected, and all rats were housed in the experimental animal center at constant temperature and humidity (temperature ~23°C, humidity ~50%) and given a 12-hour light and 12-hour dark period to simulate day and night.

[0072] 2. Experimental materials: 1×10 5N-methyl-N-nitro-N-nitrosoguanidine (MNNG) solution at a concentration of 0.1 μg / L or sterile tap water was used. For rat feed, either germ-free rat feed containing ranitidine hydrochloride at a concentration of 0.03% (hereinafter referred to as ranitidine feed) or standard feed for germ-free rats was used. For oral administration, lamb stomach extract vitamin B diluted with 50% ethanol solution or 0.9% sodium chloride solution was used. 12 The contents of the formulation or a 0.9% sodium chloride solution was used.

[0073] Construction of rat precancerous lesion model and drug intervention experiment: Rats were divided into an administration group, a non-administration group, and a negative control group, and the administration group was administered 0.2 g / kg of lamb stomach extract vitamin B once a day. 12 The formulation was orally administered to rats in the non-administered and negative control groups, and 0.2 g / kg of a 0.9% NaCl aqueous solution was orally administered once a day to rats in the non-administered and negative control groups, and the experiment was continued for three months.

[0074] Sample collection and processing: After the drug intervention experiments, the rats in the three groups were anesthetized and sacrificed after 24-36 hours of fasting and anhydrous diet. Blood samples, gastric juice samples, and gastric tissue samples were collected. The gastric tissue samples were sent to the pathology department, dehydrated, embedded in paraffin wax, sliced, and stained with hematoxylin and eosin for pathological examination.

[0075] Observation and evaluation: The general condition of the rats (coat color, usual activity, food intake, water intake, body weight change), death, pH value of gastric juice, and pathological changes in the gastric mucosa of rats in each group (observation of the macroscopic morphology of the mucosa, infiltration of inflammatory cells in the mucosa, number of hyperplastic nodules on the surface of the gastric mucosa, etc.) were observed. The pathological changes in the gastric mucosa of rats were evaluated with reference to the "Expert Consensus on the Prevention, Control and Management of Helicobacter pylori Infection in Chinese Residents' Families (2021)". The precancerous lesions of the gastric mucosa of rats in the treatment and non-treatment groups were scored (1 to 3 points), with 1 point for rats without precancerous lesions (IN), 2 points for those with LGIN, and 3 points for those with HGIN.

[0076] The results of comparing the pH of gastric juice in rats in the treatment group, non-treatment group, and negative control group are shown in Figure 1.

[0077] As shown in Figure 1, the pH of gastric juice in the treated rats was higher than that in the negative control group (3.07±0.55 vs. 2.11±0.33) and lower than that in the non-treated group (3.07±0.55 vs. 4.45±0.72). Both differences were statistically significant (t=3.45, 2.80, both P<0.05). Since gastric acid secretion was improved in the treated group, it is possible that secretory function and the type of gastric mucosal epithelial cells were also improved.

[0078] The pathological changes and precancerous scores of the gastric mucosa of rats in the treated, untreated and negative control groups were evaluated.

[0079] The gastric mucosa of the rats in the negative control group was dark orange-red in color, and no abnormal proliferation was observed on the surface (Fig. 2A); the gastric mucosa of the rats in the treatment group was similar in appearance and color to the gastric mucosa surface of the rats in the negative control group, except for a few white spots, and no obvious abnormalities were observed (Fig. 2B); the gastric mucosa surface of the fundus, body, and pyloric antrum of the rats in the untreated group contained tumor-like hyperplastic nodules of various sizes and amounts, and erosions, blood sinuses, etc. were observed, although there were individual differences (Fig. 2C). In the negative control group, normal glandular structure was observed in the gastric mucosa, and basically no infiltration of inflammatory cells or loss of glands was observed (Figure 2D); in the treated group, slight infiltration of inflammatory cells was observed in the gastric mucosa and atrophy of the glands in the lamina propria was observed, but it did not worsen compared to the end of the model period, and the density of the glands in the lamina propria of some individuals was more abundant than in the previous period (Figure 2E); in the non-treated group, atrophy of the gastric mucosa was not reversed, but rather worsened, and many atypical hyperplasias were observed (Figure 2E). From these results, it was found that lamb stomach extract vitamin B 12 can significantly reduce the risk of progression from precancerous gastric cancer to precancerous lesions, and lamb stomach extract vitamin B 12 It was suggested that this could prevent the development of precancerous lesions.

[0080] The weight gain in the treated group was greater than that in the untreated group at week 6 [(508.26±33.96)g vs. (495.50±23.01)g], and the pH value of gastric juice was lower than that in the untreated group (3.07±0.55 vs. 4.45±0.72), both of which were statistically significant (P<0.05 for both). The number of hyperplastic nodules on the gastric mucosal surface was lower in the treated group than in the untreated group at all sites [fundus 6.00 (3.00, 7.00) vs. 11.00 (7.00, 13.00)]. The scores for the pylorus were 3.00 (2.00, 4.00) vs. 0.00 (0.00, 1.00), and for the whole stomach were 15.00 (13.00, 17.00) vs. 7.00 (3.00, 10.00), and the differences were statistically significant (U = 43.50, 49.00, 49.00, all p < 0.05). The precancerous lesion score of the gastric mucosa of the treatment group was lower than that of the non-treatment group [(1.00 ± 0.00) points vs. (1.14 ± 0.38) points], and the incidence of precancerous lesions was lower than that of the non-treatment group (1 / 7 vs. 5 / 7), and the differences were statistically significant (t = 2.45, χ 2 =4.67, both P=0.031).

[0081] Lamb stomach extract-vitamin B1 was administered in rat models of chronic atrophic gastritis and intestinal metaplasia (research methods 1 and 2). 12 The preparation has been proven to promote the secretion of gastric acid and pepsin, promote the proliferation of master cells, parietal cells, G cells, D cells, regulate the secretion of neuroendocrine hormones and gastrin, growth inhibitory hormones, prostaglandin E2, epidermal growth factor and its receptors, play a role in the protection of gastric mucosa, and promote the regeneration of gastric mucosal glands, improve the bleeding volume and bleeding condition of gastric mucosa, and have obvious improvement and inhibition effects on intestinal metaplasia subtype. At the same time, it can significantly increase the level of LPOSODJSH-PX in the blood of rats, promote the proliferation of gastric mucosal gland cells, improve the synthesis of gastric mucosal cells, scavenge free radicals, play a role in the protection of gastric mucosa, and have therapeutic effects on the therapeutic reversal of chronic atrophic gastritis in rats, intestinal metaplasia, CAG.

[0082] Example 5. Vitamin B from Lamb Stomach Extract in the Treatment of Chronic Atrophic Gastritis with Intestinal Metaplasia 12Pathological effects and activity of chymosin and pepsin in preparations Background: Chronic atrophic gastritis accompanied by intestinal metaplasia is closely related to the development of gastric cancer. Lamb stomach extract Vitamin B 12 It was found that the bioactive enzymes contained in the formulation could reverse gastric mucosal atrophy and intestinal metaplasia by promoting glandular proliferation, inhibiting apoptosis, and suppressing oxidative stress in the gastric mucosa of rats.

[0083] Objective: Lamb stomach extract-vitamin B in the treatment of chronic atrophic gastritis with intestinal metaplasia 12 We will investigate the pathological effects of the biological activity of the drug and the factors that affect it.

[0084] Methods: The study included 173 patients with chronic atrophic gastritis with intestinal metaplasia who visited Saijing Hospital between June 2018 and June 2019. All patients were orally administered 3 to 6 capsules of lamb stomach 3 times per day, with an increased dose (or number of doses). Gastric endoscopy and gastric mucosal tissue biopsy were performed 6 and 12 months after the start of treatment, respectively. The treatment effect was evaluated using the changes in the OLGA stage and OLGIM stage before and after treatment, and factors affecting efficacy were explored using single-factor and multifactor analysis.

[0085] Results: After 6 months of treatment, the OLGA stage success rate of patients was 49.7%, the OLGIM stage success rate was 32.9%, and the overall success rate was 64.7%. After 12 months of treatment, the OLGA stage success rate was 56.4%, the OLGIM stage success rate was 41.8%, and the overall success rate was 70.9%.

[0086] Conclusion: Lamb stomach significantly reduced the OLGA and OLGIM stages in patients with chronic atrophic gastritis with intestinal metaplasia, suggesting that this drug may reduce the risk of gastric cancer in patients, and raising the prospect of clinical application in gastric cancer prevention.

[0087] Subjects and methods I. Research Subjects A follow-up study was conducted on 173 patients with CAG with IM who visited the gastroenterology outpatient department of Xijing Hospital, China Air Force Military Medical University from June 2018 to June 2019. Enrollment criteria were: (i) aged 18-70 years, (ii) diagnosed with CAG with IM by upper gastrointestinal endoscopy and histopathological biopsy within the past 3 months, and exclusion criteria were: (i) previously diagnosed with malignant tumor; (ii) history of gastric surgery; (iii) breastfeeding or pregnancy; (iv) endocrine or autoimmune diseases such as hypothyroidism, adrenal insufficiency, systemic lupus erythematosus, ankylosing spondylitis, and systemic vasculitis; (v) severe psychiatric illness; (vi) refusal of drug treatment; (vii) diagnosed with gastric cancer or high-grade intraepithelial neoplasia by upper gastrointestinal endoscopy and pathological examination; (viii) severe hepatic and renal failure; and (ix) incomplete clinicopathological data. Data on the patients' general condition, dietary habits, behavioral characteristics (smoking, drinking), medical history, and medication history were collected.The mean age was 54.4±8.8 years, and the Hp infection rate was 79.8% (138 / 173).

[0088] 2. Research method 1. Treatment: Lamb stomach extract vitamin B in all cases 12 The formulation (capsules, 55 U / capsule) was orally administered at 3-6 capsules / day for 12 months. At the same time, a 4-drug combination therapy including bismuth was administered to the Hp-infected patients, and the successful eradication of the bacteria was confirmed. Gastrointestinal endoscopy and mucosal histopathological examinations were repeated 6 and 12 months after the start of treatment, respectively. 2. Gastric mucosal biopsy: Biopsies were performed according to the new Sydney system histological diagnostic criteria for chronic gastritis at five sites: the greater antrum curvature, the lesser antrum curvature, the greater antrum curvature of the body of the stomach, the lesser antrum curvature of the body of the stomach, and the gastric angle. 3. Efficacy evaluation: According to the OLGA and OLGIM gastric cancer risk stage evaluation methods, the patients' gastric mucosal atrophy and IM stage were calculated, and the stage changes before treatment and 6 and 12 months after treatment were compared. If the OLGA stage and OLGIM stage decreased by 1 or more, it was judged that the treatment was effective. If any one of them was effective, it was judged that the treatment was effective overall, and if both were effective, it was judged that the treatment was effective overall.

[0089] 3. Statistical analysis The statistical software used was SPSS 22.0. The one-sample Kolmogorov-Smirnov test was used to test for normality of continuous variables, and variables that were not normally distributed were represented by M, and variables that were normally distributed were represented by .

number

[0090] result 1. Lamb stomach extract Vitamin B 12 Changes in OLGA and OLGIM stages in patients administered the drug One hundred seventy-three patients with CAG IM completed 6 months of treatment and follow-up, and 110 completed 12 months of treatment and follow-up. After 6 and 12 months of treatment, the OLGA and OLGIM stages showed a significant increase in patients with stage 0 and stage I, and a significant decrease in patients with stage II, stage III, and stage IV (Tables 11 and 12).

[0091] [Table 11]

[0092] [Table 12]

[0093] Further analysis revealed that lamb stomach extract vitamin B 12After 6 months of treatment with the preparation, the OLGA stage was reduced in 49.7% (86 / 173 cases), the OLGIM stage was reduced in 32.9% (57 / 173 cases), the OLGA and / or OLGIM stage was reduced in 64.7% (112 / 173 cases), and both the OLGA and OLGIM stages were reduced in 17.9% (31 / 173 cases) (Figure 1). 12 After 12 months of treatment with the preparation, the OLGA stage was reduced in 56.4% (62 / 110 cases), the OLGIM stage was reduced in 41.8% (46 / 110 cases), the OLGA and / or OLGIM stage was reduced in 70.9% (78 / 110 cases), and both the OLGA and OLGIM stages were reduced in 27.3% (30 / 110 cases) (Figure 2). 12 No statistically significant difference in efficacy was observed between the 6-month and 12-month treatments (P>0.05).

[0094] Before treatment, the proportion of high-risk stages (stages III and IV) was high in OLGA and OLGIM. After treatment, the proportion of high-risk stages significantly decreased, and the proportion of low-risk stages significantly increased. The difference in stage distribution after treatment compared with before treatment was statistically significant (P<0.01; Figures 3 and 4). From these results, lamb stomach extract vitamin B 12 The formulation was shown to have a significant reversal effect on atrophy and IM. Further analysis showed that after 6 and 12 months of treatment, the proportion of OLGA high-risk stages was significantly reduced compared with OLGIM stages, and the difference was statistically significant (P<0.05). Furthermore, the proportion of patients with complete disappearance of atrophy (OLGIM stage 0) and complete disappearance of IM (OLGIM stage 0) was higher after 12 months of treatment than after 6 months of treatment, but the difference was not statistically significant (P>0.05).

[0095] 2. Hp infection and vitamin B from lamb stomach extract 12 Relationship of efficacy of formulation According to the Hp infection, patients were divided into Hp negative group (patients who were always Hp negative before enrollment), Hp eradication experience group (patients who were successfully eradicated 3 months before enrollment and were judged to be negative before enrollment), and pre-treatment Hp eradication group (patients who were Hp positive at enrollment and were judged to be Hp negative before enrollment). 12 The differences in OLGA and OLGIM stage efficiency at 6 months of treatment were not statistically significant in any of the three groups (P>0.05); the differences in OLGA stage and OLGIM stage efficiency at 12 months of treatment were not statistically significant in any of the three groups (P>0.05). 12 The use of the formulation composition did not significantly affect the efficacy rate of OLGA stage, but could more effectively reduce the OLGIM stage of the patients (P<0.05; Table 13).

[0096] [Table 13]

[0097] Example 6 Lamb stomach extract Vitamin B 12 Phase IV clinical trial to treat chronic atrophic gastritis with formulation Vitamin B from lamb stomach extract for chronic atrophic gastritis 12 To evaluate the efficacy and safety of the formulation, Lamb Stomach Extract Vitamin B12 for reversing glandular atrophy in chronic atrophic gastritis. 12 The efficacy of the formulation was evaluated and pharmacoeconomic data were collected during the treatment period.

[0098] Research method A randomized, double-blind, multicenter clinical trial will be conducted using a placebo as the control and histopathological changes as the primary efficacy endpoint. Subjects who passed the screening will be randomly assigned to the test group or placebo group in a 2:1 ratio.

[0099] Evaluation criteria Treatment indicators: (1) Efficacy: Efficacy was defined as a decrease of 1 or more points in the histopathological score from baseline. For histopathological evaluation, biopsies were taken from five sites: the lesser curvature of the pyloric antrum, the greater curvature of the pyloric antrum, the angle of the stomach, the lesser curvature of the gastric body, and the greater curvature of the gastric body. Chronic inflammation, activity, atrophy, and intestinal metaplasia were classified into four stages: normal (0 points), mild (1 point), moderate (2 points), and severe (3 points).

[0100] The efficacy rate was calculated and evaluated for each of the five sites: the lesser antrum curvature, the greater antrum curvature, the gastric angle, the lesser antrum curvature of the gastric body, and the greater antrum curvature of the gastric body. Cases in which the histopathological evaluation at the time of enrollment was normal were not included in the calculation of the efficacy rate. A comprehensive evaluation of the efficacy rate was performed using the following two methods: (i) Overall evaluation 1: The scores of the most severe areas before and after treatment were selected separately to calculate the effectiveness rate; (ii) Overall evaluation 2: The sites where the subject's chronic inflammation, activity, atrophy, and intestinal metaplasia were most severe at the time of enrollment were selected as the target observation sites, and the effectiveness rate of the target observation sites 180 days after administration was evaluated. If the scores at enrollment were the same for multiple sites, the site with the smallest change in score before and after treatment was selected for evaluation. In this case, the integrated evaluation 1 method was used as the main indicator for evaluating the effectiveness rate.

[0101] (2) Reversal incidence: Reversal incidence was defined as a reduction of 2 or more points from baseline in the histopathological score after 180 days in enrolled moderate and severe patients. The incidence of reversal was calculated and evaluated at five sites: the lesser curvature of the antrum, the greater curvature of the antrum, the angle of the stomach, the lesser curvature of the gastric body, and the greater curvature of the gastric body. Cases with normal or mild histopathological evaluation at the time of enrollment were not included in the calculation of the incidence of reversal. The overall incidence of reversal was evaluated in the following two ways: (i) Overall assessment 1: The scores of the most severe sites before and after treatment were selected separately, and the incidence of reversal was calculated; (ii) Overall evaluation 2: The sites with the most severe chronic inflammation, activity, atrophy, and intestinal metaplasia at the time of registration were selected as the target observation sites, and the incidence of reversal at the target observation sites 180 days after administration was evaluated. If the scores at the time of registration were the same for multiple sites, the site with the smallest change in score before and after treatment was selected for evaluation. In this case, the method of integrated evaluation 1 was used as the main indicator for assessing the reversal occurrence rate.

[0102] (3) Histopathological score: The total histopathological score of the five sites, the lesser curvature of the pyloric antrum, the greater curvature of the pyloric antrum, the gastric angle, the lesser curvature of the gastric body, and the greater curvature of the gastric body, the most severe histopathological score of each site, and the histopathological score of each site were evaluated. In the histopathological evaluation, chronic inflammation, activity, atrophy, intestinal metaplasia, etc. were evaluated.

[0103] (4) Endoscopic gastric mucosal changes: These included endoscopic signs of atrophic gastritis (red and white mucosa, white mucosa predominance, flattened folds, loss of folds, vascularization, granular or nodular mucosa) and those with endoscopic erosions or bile reflux (flat erosions, raised erosions, hemorrhagic petechiae, mucosal congestion, bile reflux). The cardia, body, angle, antrum, fundus, and pylorus were evaluated separately.

[0104] (5) Clinical symptom score: Seven items: abdominal discomfort, loss of appetite, abdominal bloating, dyspeptic diarrhea, abdominal pressure, gastric acid reflux, and nausea / vomiting. Of these, the abdominal pain score consisted of three items: abdominal discomfort, abdominal bloating, and abdominal pressure, and the scores for items other than abdominal pain consisted of four items: loss of appetite, dyspeptic diarrhea, gastric acid reflux, and nausea / vomiting.

[0105] safety index Adverse events; Laboratory tests: routine blood tests (red blood cells, hemoglobin, white blood cells, platelets, neutrophils), blood biochemistry tests (ALT, AST, BUN, Cr, fasting blood glucose), routine urine tests (urine white blood cells, urine red blood cells, urine sugar), routine stool tests, occult blood tests; electro-cardiogram; General examination: temperature, pulse, blood pressure, respiration, etc.

[0106] Statistical methods: All statistical tests were performed using two-sided tests, and a p-value of 0.05 or less was considered to indicate statistical significance for the differences tested (unless otherwise stated). Measurement information was statistically expressed as mean, median, standard deviation, maximum, minimum, 25% quartile, and 75% quartile.

[0107] Efficacy analysis 1. Effectiveness rate of chronic inflammation by area The efficacy of lamb stomach extract with vitamin B12 in chronic inflammation in the greater antrum and greater corpus of the stomach in patients with chronic atrophic gastritis 12 The results were higher in the formulation group than in the placebo group (P<0.05). The results are shown in Figure 3.

[0108] 2. Chronic inflammation reversal incidence by area The incidence of reversal of chronic inflammation in the greater and lesser curvatures of the stomach in patients with chronic atrophic gastritis is increased by lamb stomach extract with vitamin B 12 The results were higher in the formulation group than in the placebo group (P<0.05). The results are shown in Figure 4.

[0109] 3. Effective rate of atrophy by area The effectiveness of lamb stomach extract vitamin B1 in reducing atrophy in the greater curvature of the stomach, the lesser curvature of the stomach, the angle of the stomach, the lesser curvature of the antrum, and the greater curvature of the antrum in patients with chronic atrophic gastritis was 12 The results were higher in the formulation group than in the placebo group (P<0.05). The results are shown in Figure 5.

[0110] 4. Incidence of atrophy reversal by area The incidence of reversal of atrophy in the greater curvature of the stomach, the lesser curvature of the stomach, the angle of the stomach, the lesser curvature of the antrum, and the greater curvature of the antrum in patients with chronic atrophic gastritis was significantly increased by the administration of lamb stomach extract vitamin B 12 The results of the study were higher in the formulation group than in the placebo group (P<0.05). The results are shown in Figure 6.

[0111] 5. Effectiveness rate of intestinal metaplasia by site The efficacy of lamb stomach extract vitamin B1 in the prevention of intestinal metaplasia in the lesser curvature of the stomach, the gastric angle, the lesser curvature of the antrum, and the greater curvature of the antrum in patients with chronic atrophic gastritis was12 The results were higher in the formulation group than in the placebo group (P<0.05). The results are shown in Figure 7.

[0112] 6. Incidence of intestinal metaplasia reversal by location The incidence of reversal of intestinal metaplasia in the lesser curvature of the stomach, the gastric angle, the lesser curvature of the antrum, and the greater curvature of the antrum in patients with chronic atrophic gastritis was significantly increased by the administration of lamb stomach extract with vitamin B 12 The formulation group was higher than the placebo group (P<0.05). The results are shown in Figure 8.

[0113] 7. Comprehensive evaluation of efficacy rate The efficacy of lamb stomach extract with vitamin B12 in reducing chronic inflammation, atrophy and intestinal metaplasia in patients with chronic atrophic gastritis 12 The formulation group was higher than the placebo group (P<0.05). The results are shown in Figure 9.

[0114] 8. Comprehensive evaluation of reversal occurrence rate The incidence of reversal of chronic inflammation, atrophy and intestinal metaplasia in patients with chronic atrophic gastritis was increased by the administration of lamb stomach extract with vitamin B 12 The formulation group was higher than the placebo group (P<0.05). The results are shown in Figure 10.

[0115] Efficacy Results: Effectiveness rate A comprehensive evaluation of 1 was used, i.e., the most severe of the histopathological evaluations before and after treatment was used to calculate the efficacy rate: Comparison of chronic inflammation efficacy rates between groups: efficacy rate FAS (PPS), placebo 10.60% (12.13%), lamb stomach 15.70% (17.86%), no two-way CMH-χ 2 The results of the FAS test were P=0.0411 and the PPS test was P=0.0492, and both intergroup differences were statistically significant.

[0116] Comparison of active efficacy rates between groups: Effectiveness rate FAS (PPS), placebo 34.48% (38.16%), lamb stomach 41.07% (45.58%), no two-way CMH-χ 2The results of the test were P=0.5336 for the FAS results and P=0.4601 for the PPS results, and neither of the differences between the groups was statistically significant.

[0117] Comparison of atrophy effectiveness rates between groups: Effectiveness rate FAS (PPS), placebo 7.80% (8.79%), lamb stomach 16.01% (18.60%), no two-way CMH-χ 2 The results of the FAS test were P=0.0010 and the PPS test was P=0.0008, and both intergroup differences were statistically significant.

[0118] Two-way layout without CMH-χ 2 The FAS result was P=0.0003, and the PPS result was P=0.0003, and both intergroup differences were statistically significant. Because there was a baseline imbalance in patients with the most severe intestinal metaplasia, a logistic regression model was applied with the most severe intestinal metaplasia at baseline as a covariate and the intestinal metaplasia success rate as a response variable to further examine the intestinal metaplasia success rate. As a result, the FAS result was P=0.0001, and the PPS result was P=0.0001, and the intergroup differences were still statistically significant.

[0119] In other words, the area with the highest histopathological evaluation before treatment was used to calculate the efficacy rate: A comprehensive evaluation of 2 was used, i.e., the area with the highest histopathological evaluation before treatment was used to calculate the efficacy rate: Comparison of chronic inflammation efficacy rates between groups: efficacy rate FAS (PPS), placebo 39.93% (46.44%), lamb stomach 49.21% (57.56%), no two-way CMH-χ 2 The results of the FAS test were P=0.0103 and the PPS test were P=0.0063, and both intergroup differences were statistically significant. Comparison of active efficacy rates between groups: Effectiveness rate FAS (PPS), placebo 58.62% (65.79%), lamb stomach 63.69% (70.07%), no two-way CMH-χ 2The results of the test were P=0.6068 for the FAS results and P=0.7386 for the PPS results, and neither of the differences between the groups was statistically significant. Comparison of atrophy effectiveness rates between groups: Effectiveness rate FAS (PPS), placebo 36.17% (41.84%), lamb stomach 59.61% (69.13%), no two-way CMH-χ 2 The results of the FAS test were P<0.0001, and the PPS test was P<0.0001, and both intergroup differences were statistically significant. Comparison of intestinal metaplasia efficacy rates between groups: efficacy rate FAS (PPS), placebo 34.29% (38.31%), lamb stomach 62.24% (69.25%), no two-way CMH-χ 2 The results of the FAS test were P<0.0001, and the PPS test was P<0.0001, and both intergroup differences were statistically significant.

[0120] Reversal Rate A global score of 1 was used, i.e., the most severe of the pre- and post-treatment histopathological scores, to calculate the reversal incidence: Comparison of incidence of chronic inflammation reversal between groups: No subjects in either the lamb stomach or placebo group experienced reversal. Comparison of the incidence of activity reversal between groups: incidence reversal rate FAS (PPS), placebo 33.33% (35.48%), lamb stomach 20.73% (22.54%), no two-way CMH-χ 2 The results of the test were P=0.1044 for the FAS results and P=0.3446 for the PPS results, and neither of the differences between the groups was statistically significant. Comparison of the incidence of atrophy reversal between groups: incidence reversal rate FAS (PPS), placebo 0.00% (0.00%), lamb stomach 1.13% (1.34%), no two-way CMH-χ 2 The results of the test were P=0.0818 for the FAS results and P=0.0810 for the PPS results, and neither of the differences between the groups was statistically significant. Comparison of the incidence of intestinal metaplasia reversal between groups: incidence reversal rate FAS (PPS), placebo 3.64% (4.08%), lamb stomach 13.62% (14.67%), no two-way CMH-χ 2The results of the FAS test were P=0.0079 and the PPS test were P=0.0139, and both intergroup differences were statistically significant.

[0121] Because there was a baseline imbalance in patients with the most severe intestinal metaplasia, we further examined the incidence of intestinal metaplasia reversal by fitting a logistic regression model with the most severe intestinal metaplasia at baseline as the covariate and the incidence of intestinal metaplasia reversal as the response variable.The results were P = 0.0083 for the FAS and PPS, respectively, and P = 0.0104, indicating that the difference between the groups remained statistically significant.

[0122] A comprehensive evaluation of 2 was adopted, i.e., the area with the highest histopathological evaluation before treatment was used to calculate the reversal incidence: Comparison of chronic inflammation reversal incidence between groups: incidence reversal rate FAS (PPS), placebo 1.59% (1.87%), lamb stomach 6.10% (6.68%), no two-way CMH-χ 2 The results of the FAS test were P=0.0057, and the PPS test was P=0.0082, and both intergroup differences were statistically significant. Comparison of the incidence of activity reversal between groups: incidence reversal rate FAS (PPS), placebo 63.89% (70.97%), lamb stomach 50.00% (53.52%), no two-way CMH-χ 2 The test showed that the FAS result was P=0.0424, and the difference between the groups was statistically significant; the PPS result was P=0.0626, and the difference between the groups was not statistically significant. Comparison of the incidence of atrophy reversal between groups: incidence reversal rate FAS (PPS), placebo 6.39% (7.14%), lamb stomach 19.96% (23.16%), no two-way CMH-χ 2 The FAS and PPS results were P<0.0001 and P<0.0001, respectively, and the differences between the groups were both statistically significant. Comparison of the incidence of intestinal metaplasia reversal between groups: incidence reversal rate FAS (PPS), placebo 11.82% (13.27%), lamb stomach 47.86% (53.33%), no CMH-χ 2The results of the FAS test were P<0.0001, and the PPS test was P<0.0001, and both intergroup differences were statistically significant.

[0123] Histopathological score Comparison of the sum of histopathological scores at five sites of chronic inflammation between groups: before treatment, mean ± SD, placebo FAS (PPS) 8.32 ± 2.35 (8.33 ± 2.33), lamb stomach FAS (PPS) 8.19 ± 2.28 (8.21 ± 2.22); after treatment, mean ± SD, placebo FAS (PPS) 8.63 ± 2.28 (8.72 ± 2.25), lamb stomach FAS (PPS) 8.19 ± 2.14 (8.22 ± 2.06); (post-treatment - pre-treatment) mean difference ± SD, placebo FAS (PPS) 0.31 ± 1.72 (0.38 ± 1.81), lamb stomach FAS (PPS) -0.00 ± 2.04 (0.00 ± 2.20). Paired t-test showed P = 0.0246 for FAS results and P = 0.0203 for PPS results, and both intergroup differences were statistically significant.

[0124] Comparison of the sum of histopathological scores at the five active sites between groups: pretreatment mean ± SD, placebo FAS (PPS) 0.95 ± 1.96 (0.96 ± 1.90), lamb stomach FAS (PPS) 1.01 ± 2.03 (1.02 ± 1.98); posttreatment mean ± SD, placebo FAS (PPS) 1.21 ± 2.32 (1.28 ± 2.38), lamb stomach FAS (PPS) 1.27 ± 2.35 (1.34 ± 2.37); (post-treatment - pre-treatment) mean difference ± SD, placebo FAS (PPS) 0.26 ± 2.11 (0.32 ± 2.20), lamb stomach FAS (PPS) 0.26 ± 2.13 (0.32 ± 2.28). Paired t-test showed P = 0.9909 for FAS results and P = 0.9997 for PPS results, and none of the differences between groups were statistically significant.

[0125] Comparison of the sum of histopathological scores at five sites of atrophy between groups: pretreatment mean ± SD, placebo FAS (PPS) 7.77 ± 3.05 (7.75 ± 2.98), lamb stomach FAS (PPS) 8.21 ± 3.00 (8.25 ± 3.00); posttreatment mean ± SD, placebo FAS (PPS) 9.17 ± 2.86 (9.38 ± 2.66), lamb stomach FAS (PPS) 8.75 ± 2.56 (8.89 ± 2.45); (post-treatment - pre-treatment) mean difference ± SD, placebo FAS (PPS) 1.40 ± 2.53 (1.63 ± 2.59), lamb stomach FAS (PPS) 0.54 ± 2.92 (0.63 ± 3.13). Paired t-test showed P < 0.0001 for FAS results and P < 0.0001 for PPS results, and both intergroup differences were statistically significant.

[0126] Comparison of the sum of histopathological scores in the five sites of intestinal metaplasia between groups: pretreatment mean ± SD, placebo FAS (PPS) 2.31 ± 2.75 (2.45 ± 2.79), lamb stomach FAS (PPS) 2.90 ± 3.18 (3.01 ± 3.20); posttreatment mean ± SD, placebo FAS (PPS) 2.97±3.42 (3.25±3.52), lamb stomach FAS (PPS) 2.78±3.07 (2.89±3.09); (post-treatment-pre-treatment) mean ±SD difference, placebo FAS (PPS) 0.66±1.90 (0.80±2.02), lamb stomach FAS (PPS) -0.12±2.19 (-0.13±2.36). By paired t-test, P<0.0001 for FAS and PPS, and P<0.0001 for PPS, and both intergroup differences were statistically significant.

[0127] Clinical Symptom Score The change in the total clinical symptom score at the end of treatment was not statistically significant between subgroups by analysis of covariance (FAS: P = 0.4637, PPS: P = 0.9235), but there was a consistent trend in between-group differences between centers (FAS: P = 0.8169, PPS: P = 0.9044); The adjusted mean difference (95% confidence interval) between placebo and lamb stomach was 0.24 (-0.40, 0.88) for FAS and 0.03 (-0.67, 0.74) for PPS; none of the between-group differences were statistically significant. The adjusted mean (95% CI) change in clinical score totals at the end of placebo treatment was -0.41 (-1.22, 0.40) for FAS and -0.37 (-1.35, 0.61) for PPS, neither difference was statistically significant.The adjusted mean (95% CI) change in clinical score totals at the end of lamb stomach treatment was -0.65 (-1.37, 0.07) for FAS and -0.41 (-1.32, 0.51) for PPS, neither difference was statistically significant. The change in total abdominal pain score at the end of treatment was not statistically significantly different between subgroups by analysis of covariance (FAS: P = 0.0976, PPS: P = 0.5786), and there was a consistent tendency for between-group differences between facilities (FAS: P = 0.9651, PPS: P = 0.9743); The adjusted mean difference (95% confidence interval) between placebo and lamb stomach was 0.38 (-0.07, 0.82) for FAS and 0.14 (-0.35, 0.63) for PPS; none of the between-group differences were statistically significant. The adjusted mean (95% CI) change in total abdominal pain score at the end of placebo treatment was 0.07 (-0.50, 0.63) for FAS and -0.14 (-0.81, 0.54) for PPS; neither difference was statistically significant.The adjusted mean (95% CI) change in total abdominal pain score at the end of lamb stomach treatment was -0.31 (-0.81, 0.20) for FAS and -0.27 (-0.91, 0.36) for PPS; neither difference was statistically significant. The total score change of clinical symptoms other than abdominal pain at the end of treatment was not statistically significant by analysis of covariance (FAS: P = 0.7218, PPS: P = 0.6078). The difference between the groups at the facilities showed a consistent tendency (FAS: P = 0.4674, PPS: P = 0.7934); The adjusted mean difference (95% confidence interval) between placebo and lamb stomach was −0.07 (−0.46, 0.32) for FAS and −0.11 (−0.53, 0.31) for PPS; none of the between-group differences were statistically significant. The adjusted mean (95% CI) change in the total score of clinical symptoms other than abdominal pain at the end of placebo treatment was -0.40 (-0.89, 0.09) for FAS and -0.21 (-0.80, 0.38) for PPS, neither difference was statistically significant.The adjusted mean (95% CI) change in the total score of clinical symptoms other than abdominal pain at the end of lamb stomach treatment was -0.33 (-0.77, 0.11) for FAS and -0.10 (-0.65, 0.45) for PPS, neither difference was statistically significant.

[0128] As can be seen from the above results: 1. Lamb stomach extract Vitamin B 12 The formulation is generally more effective in chronic atrophic gastritis and shows better histological improvement. 2. It is histologically effective in chronic inflammation, atrophy, and intestinal metaplasia, and is effective in reversing intestinal metaplasia.

[0129] Example 7 Real-world study using big data: Lamb stomach extract vitamin B 12 Reversal of chronic atrophic gastritis and intestinal metaplasia by pharmaceutical agents Lamb stomach extract Vitamin B 12 CAG patients treated with the formulation were analyzed before and after their own controls. 1. Treatment: Lamb stomach extract Vitamin B 12 After 6 months of regular treatment (110 U / time, 3 times / day), upper gastrointestinal endoscopy and pathological biopsy of the gastric mucosa were performed to evaluate the therapeutic effect, and a urea breath test was performed to detect the presence or absence of H. pylori infection. Treatment was started with eradication of H. pylori infection if present, and then after 2 weeks, administration of lamb stomach extract vitamin B 12 Treatment with the drug was started. 2. Endoscopic biopsy: According to the requirements of the New Sydney Standard, biopsy specimens were taken from the lesser curvature of the antrum 2-3 cm from the pylorus, the greater curvature of the antrum 2-3 cm from the pylorus, the lesser curvature of the body of the stomach 4 cm from the gastric angle, the greater curvature of the body of the stomach 8 cm from the cardia, and five biopsy specimens were taken from the gastric angle. 3. Determination of the inflammatory state and atrophy of the gastric mucosa: The inflammatory state of the gastric mucosa was evaluated using the OLGA stage system for chronic gastritis, and was divided into stages 0-IV based on the scoring level and recorded as 0-4 points. Of these, 1-2 points were defined as mild inflammation, and 3-4 points as severe inflammation. Atrophy and intestinal metaplasia were staged using the OLGA and OLGIM stage systems for chronic gastritis, respectively, and the overall evaluation results were divided into stages 0-IV, with stages 0-II defined as low stage and stages III or IV defined as high stage. 4. Efficacy index: Effective reversal of atrophy or enteritis was defined as a decrease in the OLGA or OLGIM stage after treatment by one stage or more compared to before treatment, and progression of atrophy or enteritis was defined as an increase in the OLGA or OLGIM stage after treatment by one stage or more compared to before treatment. Effective reversal rate (%) = number of cases with effective reversal after treatment / total number of cases x 100%, progression rate (%) = number of cases with progression after treatment / total number of cases x 100%, total effective reversal rate (%) = number of cases with effective reversal of atrophy or intestinal metaplasia after treatment / total number of cases x 100%.

[0130] statistical methods Statistical analysis was performed using SPSS 25.0. Measurement information was normal distribution.

number

[0131] General Information and Clinical Features Of the 240 cases, there were 146 men and 94 women, with an average age of 53.8±8.0 years and a BMI of 18.5 kg / m 2 17 cases (7.1%) were underweight or underweight, and BMI was 18.5-24.0 kg / m 2 147 cases (61.2%) were underweight or underweight, and BMI was 24.0-28.0 kg / m 266 cases (27.5%), BMI 28.0kg / m 2 10 cases (4.2%) had type A blood, 44 cases (18.3%) had type B blood, 47 cases (19.6%) had type B blood, 52 cases (21.7%) had type O blood, 22 cases (9.2%) had type AB blood, 75 cases (31.2%) had unknown blood type, 155 cases (64.6%) had a high school diploma or higher, 165 cases (68.8%) were urban residents, 56 cases (23.3%) had a family history of gastric cancer, 14 cases (5.8%) had a history of cholecystectomy, 43 cases (17.9%) had high blood pressure, 14 cases (5.8%) had diabetes, and 76 cases (31.7%) tested positive for Helicobacter pylori. The lowest gastric mucosal inflammation scores were 1 point in 4 cases (1.7%), 2 points in 158 cases (65.8%), 3 points in 65 cases (27.1%), and 4 points in 13 cases (5.4%).

[0132] Analysis of treatment effects As shown in Table 14, lamb stomach extract vitamin B 12 After 6 months of treatment with the drug, the proportion of patients with low OLGA and OLGIM stages (0-II) was higher than before treatment [62.9% (151 / 240 cases) vs. 50.8% (122 / 240 cases), 80.8% (194 / 240 cases) vs. 72.5% (174 / 240 cases)], and the differences were statistically significant in all cases (χ 2 = 7.143, 4.658, both P < 0.05). As shown in Table 15, the proportion of patients with severe inflammation in the gastric mucosa (inflammation score 3-4) was lower after treatment than before treatment [23.3% (56 / 240) vs. 32.5% (78 / 240)], and the difference was statistically significant (χ 2 = 5.011, P < 0.05). Of 10 patients with low-grade intraepithelial neoplasia at the time of initial treatment, 8 had no detectable neoplasia at the time of posttreatment review. No side effects occurred in any of the 240 patients during treatment.

[0133] [Table 14]

[0134] [Table 15]

[0135] Lamb stomach extract Vitamin B 12 After 6 months of treatment with the formulation, the reversal rates of gastric atrophy and intestinal metaplasia were 45.4% (109 / 240 cases) and 37.9% (91 / 240 cases), respectively, for a total reversal rate of 62.9% (151 / 240 cases), of which 35 cases showed complete reversal of atrophy, 15 cases showed complete reversal of intestinal metaplasia, and 8 cases showed complete reversal of both atrophy and intestinal metaplasia. Atrophy and intestinal metaplasia progressed in 21.3% (51 / 240) and 21.3% (51 / 240) of patients, respectively. The stages of atrophy and intestinal metaplasia remained unchanged in 33.3% (80 / 240) and 40.8% (98 / 240) of patients. After treatment, 53.4% ​​(63 / 118 cases) and 54.5% (36 / 66 cases) of patients with high OLGA and OLGIM stages, respectively, reversed to low stage, and 27.0% (33 / 122 cases) and 15.5% (16 / 174 cases) of patients with low OLGA and OLGIM stages, respectively, progressed to high stage. The differences in the proportions of patients with high and low OLGA and OLGIM stages before and after treatment were all statistically significant (χ 2 = 19.492, 10.013, all P < 0.05). The proportions of patients with reversal of gastric mucosal atrophy in OLGA stages I to IV were 4 / 13, 30.3% (30 / 99), 59.5% (50 / 84), and 70.6% (24 / 34), respectively. The reversal rates of intestinal metaplasia in OLGIM stages I to IV were 11.5% (7 / 61), 49.4% (44 / 89), 51.1% (24 / 47), and 13 / 19, respectively. The reversal rates of intestinal metaplasia in OLGIM stages II and II patients were relatively insignificant (P > 0.05), but the reversal rates of intestinal chemotherapy were higher in OLGIM and higher stages than in lower stages (all P < 0.05).

[0136] Analysis of influencing factors related to effectiveness As shown in Table 16, the results of univariate analysis showed that gender and vitamin supplementation were factors influencing the reversal of atrophic gastritis, while pickled vegetable intake, gastric mucosal inflammatory state, and Helicobacter pylori infection were factors influencing the reversal of intestinal metaplasia (all P<0.05). As shown in Table 17, the results of multifactorial logistic regression analysis showed that female gender, vitamin supplementation (3 times or more per week), Helicobacter pylori negativity or successful eradication, and mild inflammation (inflammation score 1-2) were associated with increased efficacy of lamb stomach extract vitamin B12 preparation, while large intake of pickled vegetables (3 times or more per week) was associated with decreased efficacy (all P<0.05).

[0137] [Table 16] JPEG2025507205000022.jpg221170JPEG2025507205000023.jpg75170

[0138] [Table 17]

[0139] As shown in Table 18, the results of the stratified analysis showed that the differences in the atrophy reversal rate and intestinal metaplasia reversal rate between the H. pylori eradication group (76 cases) and the H. pylori-negative, uneradicated group (164 cases) were not statistically significant (all P>0.05); the total effective reversal rate in the H. pylori eradication group was higher than that in the H. pylori-negative, uneradicated group, and the difference was statistically significant (χ = 5.388, P<0.05); in the H. pylori eradication group, the comparison of the total effective reversal rate, atrophy reversal rate, and intestinal metaplasia reversal rate between eradication success and eradication failure patients was not statistically significant (all P>0.05); in the H. pylori-negative, uneradicated group, the comparison of the total effective reversal rate, atrophy reversal rate, and intestinal metaplasia reversal rate between retest-negative and retest-positive patients was not statistically significant (all P>0.05).

[0140] [Table 18]

[0141] As shown in Table 19, patients with complete reversal of atrophy had a lower rate of consuming pickled foods, and a higher rate of consuming fruit, having a higher OLGA stage, and undergoing H. pylori eradication than patients with progression, and the differences were statistically significant (χ 2 = 9.701, 40.518, 4.575, and 4.023, all P < 0.05). The proportions of alcohol intake, pickled food intake, and high OLGA values ​​were lower in patients with complete reversal of intestinal metaplasia than in patients with progressive disease, and the differences were statistically significant in all cases (χ 2 = 5.315, 3.958, and 40.518, all P<0.05). No statistically significant differences were observed between the atrophy / intestinal metaplasia complete reversal group and the progression group in terms of gender, age, family history of gastric cancer, smoking history, high salt diet, vegetable intake, and OLGIM stage (all P>0.05).

[0142] [Table 19] JPEG2025507205000027.jpg115170

[0143] As a result, the lamb stomach extract of the present invention contains vitamin B 12 The overall efficacy rate of the formulation in treating atrophy and intestinal metaplasia was 62.9%, which was comparable to the results of previous studies. Furthermore, in the present invention, risk stratification of CAG patients was performed using the OLGA and OLGIM stage systems, and the results showed that lamb stomach extract vitamin B 12 The formulation was found to be highly effective, with efficacy in up to approximately 70% of stage IV patients. The treatment reversed intestinal metaplasia from high to low OLGIM stage in 54.5% (36 / 66 patients), and lamb stomach extract vitamin B 12 It was suggested that treatment with the formulation could reduce the risk of gastric cancer in CAG patients.

[0144] Helicobacter pylori infection is the most important cause of gastric cancer, and the continuous activation and intracellular positive feedback of inflammation-cancer pathways such as nuclear factor kappa B, signal transducer and activator of transcription 3 under the infection state are one of the important risk factors for the irreversible progression of gastric mucosal lesions. Helicobacter pylori eradication can effectively block the inflammation-cancer pathway and significantly reduce the incidence of gastric cancer in the population. Even after Helicobacter pylori eradication, intestinal metaplasia remains an important risk factor for the development and progression of gastric cancer. According to several studies and meta-analyses, Helicobacter pylori eradication can only reverse gastric mucosal atrophy, but cannot reverse intestinal metaplasia. Therefore, in the Correa model of gastric carcinogenesis, intestinal metaplasia is often regarded as an irreversible point of gastric mucosal lesions. The present invention and other previous studies have confirmed that gastric mucosal atrophy and intestinal metaplasia can be reversed by drug intervention, indicating that it is not appropriate to regard intestinal metaplasia as an irreversible point of gastric mucosal lesions. Furthermore, after 5-10 years of follow-up, reversal of intestinal metaplasia was observed in 33.9% (75 / 221 cases) of the pyloric antrum and 44.4% (63 / 142 cases) of the gastric body in the H. pylori eradication group, suggesting that although H. pylori eradication can reverse atrophy and intestinal metaplasia to some extent, the reversal effect is weak and requires the accumulation of a longer period of time. 12 The formulation was effective in preventing atrophy and intestinal metaplasia in patients with or without Helicobacter pylori infection, and eradication therapy was associated with increased efficacy of lamb stomach extract and vitamin B 12 The overall reversal effect of the formulation on atrophy and intestinal metaplasia was shown to be increased.

[0145] In conclusion, lamb stomach extract vitamin B 12 Treatment with the preparation may have a better reversal effect on atrophy and intestinal metaplasia in CAG patients, and may reduce the risk of gastric cancer. For CAG patients at high risk (OLGA, OLGIM stage III and IV), lamb stomach extract vitamin B 12 Pharmaceutical treatment should be recommended.

[0146] Example 8 Lamb stomach extract Vitamin B 12Reversal of chronic atrophic gastritis with severe intestinal metaplasia by a pharmaceutical preparation The patient was a 52-year-old woman who was admitted to the outpatient gastroenterology department of Xijing Hospital, China Air Force Military Medical University on March 13, 2018, complaining of "a worsening of epigastric discomfort, including abdominal bloating and borborygmus, over the past month."

[0147] 13C-UBT was negative, and gastroscopy revealed that the gastric mucosa was less smooth, with obvious intervals in the gastric body, the submucosal vascular lines were visible, and scattered punctate desquamation erosions and old bleeding spots were observed, as shown in Figure 11. The gastric body, gastric angle, and pyloric antrum were mostly gray-white microemboli, with some of their surfaces slightly depressed and reddened. Narrow-band endoscopic observation revealed bright blue crista-like changes, most of which were intestinal metaplasia. Samples were taken from the greater and lesser curvatures of the upper gastric body, the gastric angle, and the greater and lesser curvatures of the pyloric antrum. The tissues were soft, peristaltic, and normal, and the mucus lake was slightly turbid. Pathological findings, as shown in Figure 12, suggested moderate mucosal chronic inflammation, severe atrophy, and severe intestinal metaplasia in the greater and lesser curvatures of the upper gastric body, and moderate mucosal chronic inflammation, moderate atrophy, and mild intestinal metaplasia in the gastric angle and the greater and lesser curvatures of the pyloric antrum. The OLGA / OLGIM stage of gastritis based on intestinal metaplasia was IV / III. Diagnosis: chronic atrophic gastritis with severe intestinal metaplasia. The patient had epigastric discomfort, including abdominal distension and borborygmus, and was treated with lamb stomach extract vitamin B. 12The preparation was administered orally. After half a year of treatment, 13C-UBT was repeated on December 11, 2019, but the result was negative. According to the results of gastroscopy (Fig. 13), the greater curvature of the lower part of the cardia showed shallow depressions of flaky mucosa, the bottom was not mossy, the mucosal edge was still smooth and slightly congested, the tissue of the sampling site was soft, the lower part of the lesser curvature of the gastric body showed a slight protuberance of flaky mucosa, the tip of small blisters was visible, the tissue of the sampling site was soft, the remaining part of the gastric mucosa was still smooth, soft, reddish-white in color, the pyloric antrum dotted with flaky mucosa was erythematous, peristaltic movement was possible, and the mucus lake was slightly turbid. The pathological biopsy results on December 11, 2019 are shown in Figure 14. The results are: moderate chronic inflammation of the lower gastric mucosa with mild acute activity, moderate atrophy, moderate intestinal metaplasia, and microconvex hyperplasia; chronic inflammation of the cardiac mucosa with microconvex hyperplasia and moderate intestinal metaplasia; mild chronic inflammation of the pyloric antrum and gastric angle mucosa with mild acute activity and mild to moderate atrophy; mild chronic inflammation of the upper gastric mucosa. The OLGA / OLGIM stage is III / II. Diagnosis: chronic atrophic gastritis with intestinal metaplasia. Gastric mucosal atrophy and intestinal metaplasia have improved from previous cases, and lamb stomach extract vitamin B 12 The patient was instructed to continue taking the preparation orally and be re-examined in 6 months. On March 30, 2021, the 13C-UBT re-examination was negative. The results of gastroscopy, as shown in Figure 15, suggested localized mucosal congestion in the epigastric region, the remaining gastric mucosa was not soft and smooth, the small curvature on the gastric body side was obvious, the red and white colors were obvious, the submucosal vascular lines were visible, peristalsis was normal, and the mucus was slightly cloudy. Samples were taken from the greater and lesser curvatures of the gastric body, the gastric angle, and the greater and lesser curvatures of the pyloric antrum, respectively, and the tissues were soft. The pathological biopsy results (Figure 16) suggested moderate chronic inflammation, mild acute activity, mild atrophy, and small concave epithelial hyperplasia in the mucosa of the pyloric antrum and gastric angle. The mucosa of the gastric body had mild chronic inflammation. The OLGA / OLGIM stage was stage I / stage 0. Diagnosis: chronic atrophic gastritis, myocarditis. After 1.5 years of treatment with lamb stomach extract Vibramycin B preparation, the patient's OLGA stage of the gastric mucosa was reduced from stage IV to stage I, and the OLGIM stage was reduced from stage III to stage 0. Lamb stomach extract Vitamin B 12The formulation effectively reduced the OLGA and OLGIM stages in patients with CAG with intestinal metaplasia, suggesting that it may reduce the risk of gastric cancer in these patients.

[0148] It should be noted that what has been described above are preferred embodiments of the present invention, and those skilled in the art can make many improvements and modifications without departing from the principles described in the present invention.

Claims

1. Lamb stomach extract Vitamin B １２ A pharmaceutical composition comprising a lamb stomach extract and vitamin B １２ The lamb stomach extract has a chymosin titer of more than 40 U / g and a pepsin titer of more than 15 U / g, and is a mixture of lamb stomach extract and vitamin B １２ The weight ratio of the pharmaceutical composition to the pharmaceutical composition is 100,000:0.5-5.

2. The lamb stomach extract and vitamin B １２ The lamb stomach extract vitamin B according to claim 1, characterized in that the weight ratio of １２ Pharmaceutical compositions.

3. The lamb stomach extract and vitamin B １２ 3. The lamb stomach extract vitamin B according to claim 2, wherein the weight ratio of １２ Pharmaceutical compositions.

4. The lamb stomach extract vitamin B according to any one of claims 1 to 3, characterized in that the chymosin titer is 45 U / g or more and the pepsin titer is 20 U / g or more; or the chymosin titer is 55 U / g or more and the pepsin titer is 20 U / g or more. １２ Pharmaceutical compositions.

5. A lamb stomach extract vitamin B according to any one of claims 1 to 3, characterized in that the proteins of the lamb stomach extract have a relative molecular weight of 10,000 to 45,000. １２ Pharmaceutical compositions.

6. A lamb stomach extract vitamin B according to any one of claims 1 to 3, characterized in that the ratio of the chymosin titer to the pepsin titer is between 2.5 and 5.

1. １２ Pharmaceutical compositions.

7. The lamb stomach extract vitamin B according to any one of claims 1 to 6, further comprising a pharma- ceutically acceptable additive. １２ Pharmaceutical compositions.

8. The lamb stomach extract vitamin B according to claim 7, characterized in that the pharma- ceutically acceptable additive is one or more of an excipient, a binder, a diluent, a flow aid, a flavor enhancer, an antioxidant, and a bacteriostatic agent. １２ Pharmaceutical compositions.

9. The lamb stomach extract vitamin B according to any one of claims 1 to 6, wherein the formulation is in the form of a capsule, tablet, suspension or granule. １２ Pharmaceutical compositions.

10. Lamb stomach extract vitamin B in the manufacture of a medicament for reversing gastric mucosal atrophy １２ Use of the pharmaceutical composition.

11. Lamb stomach extract vitamin B in the manufacture of a medicament for treating moderate or severe chronic atrophic gastritis １２ Use of the pharmaceutical composition.

12. Lamb stomach extract vitamin B in the manufacture of a medicament for treating intestinal metaplasia １２ Use of the pharmaceutical composition.

13. The use according to claim 12, wherein the intestinal metaplasia is chronic atrophic gastritis with intestinal metaplasia.

14. Lamb stomach extract with vitamin B in the manufacture of a medicine for treating gastric intraepithelial neoplasia １２ Use of the pharmaceutical composition.

15. Lamb stomach extract vitamin B in the manufacture of a medicine for preventing stomach cancer １２ Use of the pharmaceutical composition.

16. The use according to any one of claims 10 to 14, characterized in that the treatment is an effective treatment.

17. Use according to any one of claims 10 to 14, characterized in that the treatment is a reversal treatment.

18. The use according to any one of claims 10 to 14, characterized in that the subject of administration is a patient who is negative for Helicobacter pylori.

19. Use according to any one of claims 10 to 14, characterized in that the treatment, reversal or prevention is achieved via antioxidant pathways.

20. The use according to any one of claims 10 to 14, characterized in that the treatment, reversal or prevention is achieved by regulating dysbiosis of the intestinal microflora.

21. 21. The use according to any one of claims 8 to 20, characterized in that the lamb stomach extract in the pharmaceutical composition has a chymosin titer of 45 U / g or more and a pepsin titer of 20 U / g or more; or a chymosin titer of 55 U / g or more and a pepsin titer of 20 U / g or more, is administered to the patient at 330 U, three times a day, and the proteins of the lamb stomach extract have a relative molecular weight of 10,000 to 45,000.

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