Application of antioxidant alpha-lipoic acid in preparation of medicine for improving gastritis

Abstract

The invention relates to the technical field of medicines, and particularly discloses application of an antioxidant alpha-lipoic acid in preparation of a medicine for improving gastritis. Chronic atrophic gastritis (CAG) is closely related to intestinal gastric cancer, wherein spasmolysis polypeptide expression metaplasia (SPEM) is a key node of pathological progress. Alpha-lipoic acid (ALA) is used for treating an SPEM mouse model, and the ALA is found to reduce lipid peroxidation and alleviate ferroptosis by activating an NRF2 antioxidant pathway, and significantly relieve SPEM symptoms and excessive generation of proinflammatory cytokines. According to the research, the protective effect of ALA on mouse SPEM is determined, and a new improvement choice is provided for improvement and regulation of SPEM. ALA, as a naturally existing antioxidant and anti-inflammatory agent, has a broad market prospect and social value in new application in protection of SPEM. The invention is expected to open up a new research direction for prevention and treatment of chronic atrophic gastritis.

Description

Technical Field

[0001] The invention relates to the field of drug preparation, and in particular to application of an antioxidant alpha-lipoic acid in the preparation of a drug for improving gastritis. Background Art

[0002] Gastric cancer (GC) is the fifth most common malignancy in the world and the fourth leading cause of cancer-related death. Chronic atrophic gastritis (CAG), a chronic inflammatory disease characterized by a reduction or loss of gastric mucosal glands, with or without intestinal epithelial or pyloric gland metaplasia, is closely related to the occurrence of intestinal type gastric cancer. In the pathological progression of CAG, metaplasia is an important control node and the key to the early prevention and control of intestinal type gastric cancer. However, there is currently no effective way to prevent and control CAG metaplasia.

[0003] Metaplasia usually includes two forms: intestinal metaplasia (Intestinal Metaplasia) characterized by intestinal goblet cells and spasmolytic polypeptide-expressing metaplasia (SPEM) characterized by deep gastric antral glands expressing spasmolytic polypeptides. Among them, SPEM plays a vital role in the entire development of gastric cancer. Many studies have found that the appearance of SPEM precedes intestinal epithelial hyperplasia. Therefore, effectively curbing the pathological formation of SPEM is of great significance for the early prevention and treatment of intestinal gastric cancer, which is a hot spot and difficulty in this research field.

[0004] Chinese patent document CN1139403090A discloses a simulation method of chronic atrophic gastritis lesions with high simulation degree, stable and lasting, specific phenotype and high efficiency. The method uses gene editing technology to hybridize GRIM19flox / flox gene editing mice with ATP4B-Cre gene editing mice to obtain gastric mucosa specific parietal cell GRIM-19 knockout mouse strain (GRIM19- / - / ATP4B-Cre, SPF level). This gene knockout mouse has early lesions of chronic atrophic gastritis intestinal metaplasia and spontaneous spasmolytic polypeptide expression metaplasia, and can be stably expressed, not repeatedly, so it can be used as a mouse chronic atrophic gastritis pathological model for research.

[0005] On the other hand, α-lipoic acid (ALA, α-Lipoic acid, 1,2-dithiolane-3-pentanoic acid) is a naturally occurring sulfhydryl compound found in almost all plant and animal species. As a potent antioxidant and anti-inflammatory agent, ALA is able to scavenge ROS and maintain a healthy cellular redox state. However, although ALA exhibits significant antioxidant and anti-inflammatory effects in a variety of disease models, such as anti-inflammatory effects by inhibiting pathways such as NF-κB, COX-2, and STAT3, and improving mitochondrial function by enhancing antioxidant capacity and inhibiting specific pathways, its role in protecting SPEM and its mechanism are still unclear.

[0006] Specifically, although some studies have reported the protective effects of ALA in diseases such as ulcerative colitis, liver damage, toxicity caused by iron overload, and unexplained recurrent spontaneous abortion, and inhibited ferroptosis through various mechanisms such as upregulating the PPARγ / NRF2 / GPX4 signaling pathway and regulating the Keap1-Nrf2 signaling pathway, there are currently no reports on the effects of antioxidants and their derivatives in chronic atrophic gastritis, especially research on the protective effect of ALA on SPEM and its mechanism is still blank.

[0007] In order to solve the above problems, the applicant proposes the use of an antioxidant α-lipoic acid in the preparation of a drug for improving gastritis. Summary of the invention

[0008] The purpose of the present invention is to provide an application of an antioxidant alpha-lipoic acid in the preparation of a medicament for improving gastritis, so as to solve the problems in the prior art.

[0009] To achieve the above object, the present invention provides the following technical solution: a pharmaceutical composition for improving gastritis, characterized in that it contains an effective amount of α-lipoic acid as an active ingredient.

[0010] Optionally, the gastritis is chronic atrophic gastritis.

[0011] Optionally, the α-lipoic acid reduces lipid peroxidation, alleviates ferroptosis, and alleviates spasmolytic polypeptide expression metaplasia symptoms by activating the NRF2 antioxidant pathway.

[0012] Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0013] A method for improving gastritis, comprising administering an effective amount of the pharmaceutical composition according to any one of claims 1 to 4 to an individual in need thereof.

[0014] Optionally, the gastritis is chronic atrophic gastritis.

[0015] Optionally, the improvement alleviates gastritis symptoms, including reducing lipid peroxidation, reducing ferroptosis, and reducing spasmolytic polypeptide expression and metaplasia symptoms.

[0016] Beneficial effects: Innovative application in the improvement of chronic atrophic gastritis: This invention is the first to apply α-lipoic acid (ALA) to the improvement of chronic atrophic gastritis (CAG), especially its key pathological node - spasmolytic polypeptide expressing metaplasia (SPEM), filling the research gap in this field.

[0017] Clear protective mechanism: Through experimental research, the present invention reveals that ALA reduces ferroptosis by activating the NRF2 antioxidant pathway and reducing lipid peroxidation, and significantly alleviates SPEM symptoms and excessive production of proinflammatory cytokines. This discovery provides an important basis for understanding the mechanism of action of ALA in improving gastritis.

[0018] Potential clinical application prospects: In view of the natural existence of ALA and its good antioxidant and anti-inflammatory properties, the present invention provides a new improvement option for the improvement and regulation of SPEM, and is expected to open up a new research direction for the prevention and treatment of chronic atrophic gastritis, with important clinical application potential and social value. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a graph showing the viability of GES-1-134 cells after intervention with the antioxidant ALA.

[0020] Figure 2 Figure 2 is the detection of NRF2-related proteins in the antioxidant pathway of GES-1-134 cells and GRIM19- / - mouse gastric mucosal tissue after ALA intervention; A is the detection of antioxidant ALA by Western blotting ( 0 μm、 20 μm、 40 μm ) was used to detect the expression levels of NRF2 pathway marker proteins (NRF2, P-NRF2, HO-1) in GES-1-134 cells treated with the antioxidant ALA; B was used to detect the expression levels of NRF2 pathway marker proteins (NRF2, P-NRF2, HO-1) in the gastric mucosal tissue of GRIM19- / - mice treated with the antioxidant ALA.

[0021] Figure 3The figure shows the protein detection of iron death markers in the gastric mucosa tissue of GES-1-134 cells and GRIM19- / - mice after ALA intervention; A is the expression level of iron death markers (TFR, SLC7A11, GPX4, FTH1) in GES-1-134 cells treated with antioxidant ALA by immunoblotting; B is the expression level of iron death markers (TFR, SLC7A11, GPX4, FTH1, COX2) in the gastric mucosa tissue of GRIM19- / - mice treated with antioxidant ALA by immunoblotting. ;

[0022] Figure 4 This is the Lillie iron (II) staining of the gastric mucosal tissue of GRIM19- / - mice after ALA intervention;

[0023] Figure 5 To detect the expression of inflammatory factors in gastric mucosal tissue of GRIM19- / - mice after ALA intervention;

[0024] Figure 6 The SPEM pathological process of GRIM19- / - mouse gastric mucosal tissue after ALA intervention was detected and analyzed. A is the protein immunoblotting detection of gastric mucosal tissue SPEM markers TFF2, Mist1, Clustein-1, and HE4 protein levels; B is the immunofluorescence detection of GIF, GSII and Mist1 immunofluorescence triple-label analysis. (Bar: 50 μm )(***<0.001). DETAILED DESCRIPTION

[0025] The following describes the preferred embodiments of the present invention with reference to the drawings in the specification, so that the technical content is clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0026] In the drawings, components with the same structure are indicated by the same numerical reference numerals, and components with similar structures or functions are indicated by similar numerical reference numerals. The size and thickness of each component shown in the drawings are arbitrarily shown, and the present invention does not limit the size and thickness of each component. In order to make the illustration clearer, the thickness of the components is appropriately exaggerated in some places in the drawings.

[0027] The present invention provides an application of an antioxidant alpha-lipoic acid in the preparation of a drug for improving gastritis, especially for chronic atrophic gastritis and its related symptoms, such as improvement of spasmolytic polypeptide expressing metaplasia (SPEM). ALA significantly alleviates gastritis symptoms and excessive production of proinflammatory cytokines by activating the NRF2 antioxidant pathway, reducing lipid peroxidation, alleviating ferroptosis, and alleviating SPEM symptoms.

[0028] The pharmaceutical composition of the present invention comprises an effective amount of alpha-lipoic acid (ALA) as an active ingredient. ALA is an effective antioxidant and anti-inflammatory agent, which can protect gastric mucosa and alleviate gastritis symptoms through multiple mechanisms.

[0029] The pharmaceutical composition may also contain a pharmaceutically acceptable carrier or excipient to ensure the stability, solubility and bioavailability of the drug. These carriers or excipients include, but are not limited to, fillers (such as lactose, microcrystalline cellulose), binders (such as pregelatinized starch, hydroxypropyl methylcellulose), lubricants (such as magnesium stearate, talc), disintegrants (such as cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose), etc.

[0030] Pharmaceutical compositions can be prepared into a variety of dosage forms to suit different routes of administration and patient needs. Common dosage forms include oral preparations (such as tablets, capsules, granules, oral liquids), injection preparations (such as injections, lyophilized powder injections) and topical preparations (such as sprays, gels), etc.

[0031] Prepare high-quality alpha-lipoic acid raw materials to ensure their purity and stability. At the same time, prepare the required pharmaceutically acceptable carriers or excipients and perform necessary pretreatment on them.

[0032] Alpha-lipoic acid is mixed with a carrier or excipient in a certain proportion and uniformly mixed. If necessary, an appropriate amount of a binder and a lubricant may be added to facilitate the granulation and tableting process. The mixture is then made into granules by wet granulation or dry granulation technology.

[0033] The obtained granules are compressed into tablets. The tablets can be coated as required to improve their stability, mask the bitter taste or improve the appearance.

[0034] For sterile preparations such as injections, the drug solution needs to be filled into sterile containers and undergo strict sterilization to ensure the sterility and safety of the product.

[0035] ALA can activate the NRF2 antioxidant pathway and enhance the antioxidant capacity of cells. NRF2 is an important transcription factor that can regulate the expression of a variety of antioxidant enzymes and detoxification enzymes, thereby protecting cells from oxidative stress damage. By activating the NRF2 pathway, ALA can reduce the oxidative stress level of gastric mucosal cells and alleviate gastritis symptoms.

[0036] Lipid peroxidation is an important damage mechanism caused by oxidative stress. ALA can scavenge free radicals and reduce the generation of lipid peroxidation products, thereby protecting gastric mucosal cells from oxidative damage.

[0037] Ferroptosis is a new type of programmed cell death, which is closely related to the occurrence and development of many diseases. ALA can alleviate ferroptosis through various mechanisms, including inhibiting the accumulation of iron ions and reducing the generation of lipid peroxidation products, thereby protecting gastric mucosal cells from damage caused by ferroptosis.

[0038] SPEM is an important pathological feature of chronic atrophic gastritis and is closely related to the occurrence of intestinal gastric cancer. ALA can alleviate SPEM symptoms through the above mechanisms, including reducing the number of metaplastic cells and inhibiting the expression of inflammatory factors. It can delay the progression of chronic atrophic gastritis and reduce the risk of intestinal gastric cancer.

[0039] By constructing an animal model of chronic atrophic gastritis, the effects of ALA on gastritis symptoms and SPEM pathological characteristics were evaluated. The experimental results showed that ALA can significantly alleviate gastritis symptoms, including reducing gastric mucosal damage and lowering the level of inflammatory factors. At the same time, ALA can also inhibit the development of SPEM pathological characteristics, including reducing the number of metaplastic cells and inhibiting the expression of inflammatory factors.

[0040] The cell constructed by the present invention is a GES-1-134 cell model, and the GES-1-134 cell model is a GRIM19 knockout human gastric mucosal epithelial GES-1 cell (GES-1-134) monoclone.

[0041] The effects of ALA on cell antioxidant capacity, lipid peroxidation level and ferroptosis were evaluated by culturing GES-1-134 cell models in vitro. The experimental results showed that ALA could significantly improve the antioxidant capacity of cells, reduce the generation of lipid peroxidation products, and alleviate the occurrence of ferroptosis.

[0042] The safety of ALA was evaluated by using the cell drug toxicity test CCK8. The experimental results showed that ALA had low acute toxicity and no obvious side effects.

[0043] The main efficacy is that the SPEM index is significantly reduced, which effectively inhibits the pathological process of chronic atrophic gastritis. It can be used to improve chronic atrophic gastritis, effectively reduce the expression of inflammatory factors such as NLRP3, IL-33, IL-Iβ in gastric mucosal tissue, and inhibit the pathological process of ferroptosis of gastric mucosal parietal cells. ALA treats GES-1-134 cells in vitro, increases GPX4 levels, inhibits the pathological process of ferroptosis, and reduces the expression of inflammatory factors. .

[0044] SPSS and other statistical software were used for data analysis to compare the differences in the primary and secondary efficacy indicators between the ALA improvement group and the placebo control group. Statistical analysis was performed using t-test or chi-square test, with P < 0.05 considered statistically significant.

[0045] The present invention provides an application of an antioxidant alpha-lipoic acid in the preparation of a drug for improving gastritis, especially for the improvement of chronic atrophic gastritis and its related symptoms. ALA significantly alleviates gastritis symptoms and excessive production of proinflammatory cytokines by activating the NRF2 antioxidant pathway, reducing lipid peroxidation, alleviating ferroptosis, and alleviating SPEM symptoms. Clinical trial results further confirmed the effectiveness and safety of ALA in improving patients with chronic atrophic gastritis. Therefore, the present invention provides a new and effective drug option for the improvement of chronic atrophic gastritis.

[0046] Example 1: Preparation of ALA tablets

[0047] Weigh 10 g of high-quality α-lipoic acid raw material, 80 g of lactose, 50 g of microcrystalline cellulose, 10 g of pregelatinized starch, and 1 g of magnesium stearate.

[0048] Alpha-lipoic acid, lactose, microcrystalline cellulose and pregelatinized starch are mixed evenly, and an appropriate amount of purified water is added as a binder to form granules through a wet granulation technique. After the granules are dried, magnesium stearate is added as a lubricant and mixed evenly.

[0049] The uniformly mixed granules are placed in a tablet press for tableting, and the parameters of the tablet press such as pressure and time are adjusted to obtain ALA tablets with moderate hardness and smooth appearance.

[0050] The quality of the ALA tablets is checked, including appearance, hardness, disintegration time, content uniformity, etc. After ensuring that the tablets meet the quality standards, they are packaged and stored.

[0051] Example 2: Preparation of ALA injection

[0052] Weigh 5 g of high-quality α-lipoic acid raw material and an appropriate amount of water for injection.

[0053] Add α-lipoic acid raw material to an appropriate amount of water for injection, heat and stir until completely dissolved, adjust the pH value of the solution to an appropriate range (such as 6.0-7.0), and filter and sterilize with a 0.22 μm microporous filter membrane.

[0054] Fill the filtered ALA solution into sterile ampoules with an appropriate amount per bottle. Perform strict sterilization on the filled ampoules (such as steam sterilization at 121°C for 30 minutes) to ensure the sterility and safety of the product.

[0055] The ALA injection is subjected to quality inspection, including appearance, pH value, sterility inspection, content determination and other indicators. After ensuring that the injection meets the quality standards, it is packaged and stored.

[0056] Example 3: Clinical trial of ALA in improving patients with chronic atrophic gastritis

[0057] A multicenter, randomized, double-blind, placebo-controlled clinical trial was designed. Patients with chronic atrophic gastritis diagnosed by gastroscopy and pathological examination were enrolled, with a total of 200 cases. The patients were randomly divided into an ALA improvement group and a placebo control group, with 100 cases in each group. Patients in the improvement group took ALA tablets (containing 200 mg of ALA) orally daily, and patients in the control group took placebo tablets orally daily, and the treatment course was 8 weeks.

[0058] The main efficacy indicators include the improvement of gastritis symptoms (such as stomach pain, bloating, acid reflux, etc.), the reduction of gastric mucosal inflammation (such as gastric mucosal congestion, edema, erosion, etc. under gastroscopy), and the improvement of SPEM pathological characteristics (such as the reduction of metaplastic cells and the reduction of inflammatory factor levels, etc.). Secondary efficacy indicators include patients' quality of life scores and the incidence of adverse reactions.

[0059] Gastroscopy and pathological examination were performed on the patients before and after improvement to evaluate the improvement of gastritis symptoms and SPEM pathological characteristics. At the same time, the quality of life scores and adverse reactions of the patients were collected. SPSS and other statistical software were used for data analysis to compare the differences in the main efficacy indicators and secondary efficacy indicators between the ALA improvement group and the placebo control group.

[0060] The results showed that the gastritis symptoms of the patients in the ALA improvement group were significantly improved after the improvement, the degree of gastric mucosal inflammation was significantly reduced, and the SPEM pathological characteristics were also improved. Compared with the placebo control group, the symptom scores of stomach pain, bloating, acid reflux and other symptoms in the improvement group were significantly reduced (P<0.05), the pathological changes such as gastric mucosal congestion, edema, and erosion were significantly reduced (P<0.05), the number of metaplastic cells decreased (P<0.05), and the level of inflammatory factors decreased (P<0.05). At the same time, the quality of life score of the patients in the improvement group was significantly improved (P<0.05), and the incidence of adverse reactions was low and all were mild reactions.

[0061] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the attached claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0062] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A pharmaceutical composition for improving gastritis, characterized in that Contains an effective amount of alpha-lipoic acid as an active ingredient.

2. The pharmaceutical composition according to claim 1, wherein the gastritis is chronic atrophic gastritis.

3. The pharmaceutical composition according to claim 1 or 2, wherein the α-lipoic acid reduces lipid peroxidation, alleviates ferroptosis, and alleviates spasmolytic polypeptide expressing metaplasia (SPEM) symptoms by activating the NRF2 antioxidant pathway.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient, the carrier or excipient including but not limited to one or more of a filler, a binder, a lubricant, and a disintegrant.

5. A method for improving gastritis, characterized in that An effective amount of the pharmaceutical composition according to any one of claims 1 to 4 is administered to an individual in need thereof. The method according to claim 5 , wherein the gastritis is chronic atrophic gastritis.

7. The method according to claim 5 or 6, wherein the improvement alleviates gastritis symptoms, including reducing lipid peroxidation, reducing ferroptosis, and reducing spasmolytic polypeptide expressing metaplasia (SPEM) symptoms.

8. The method according to any one of claims 5 to 7, wherein the pharmaceutical composition is administered orally, and the dosage of the α-lipoic acid is 70-80 mg / kg body weight per day, and the drug composition is α-lipoic acid with a purity of 99%, which is dissolved in DMSO to prepare a storage solution (DMSO concentration is less than 10%), and then diluted with physiological saline to prepare a working solution for administration.

9. The method according to claim 8, wherein the α-lipoic acid is administered in the form of a tablet or capsule, and the tablet or capsule further comprises a pharmaceutically acceptable carrier or excipient.

10. A method for preparing the pharmaceutical composition according to any one of claims 1 to 4, characterized in that The invention comprises mixing an effective amount of alpha-lipoic acid with a pharmaceutically acceptable carrier or excipient and preparing the mixture into a dosage form suitable for administration.

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