Application of beta-alanine in preparation of medicine for preventing and / or treating uric acid metabolic disorder related diseases

Abstract

The invention discloses application of beta-alanine in preparation of a medicine for preventing and / or treating uric acid metabolic disorder related diseases. The invention provides a theoretical source of beta-alanine for reducing the uric acid level. In the earlier stage, the applicant of the invention adopts non-target metabolome and targeted metabolome data to construct a hyperuricemia prediction model, and Rmcorr correlation analysis shows that high beta-alanine level and low uric acid level have strong correlation. On the other hand, beta-alanine acid is adopted to intervene in a hyperuricemia mouse, it is proved that beta-alanine can reduce the blood uric acid level of the hyperuricemia mouse and promote urine uric acid excretion, and beta-alanine can be used for preventing and / or treating uric acid metabolic disorder related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to the use of β-alanine in the preparation of a drug for preventing and / or treating diseases related to uric acid metabolism disorders. Background Art

[0002] The core causes of hyperuricemia (HUA) lie in abnormal purine metabolism, increased uric acid production, and reduced uric acid excretion. According to epidemiological research data, currently, approximately one-fifth of the global population is suffering from HUA. This disease not only leads to disorders in human metabolism but is also regarded as a potential risk factor for various diseases such as gout, obesity, type 2 diabetes, hypertension, cardiovascular and cerebrovascular diseases, and secondary chronic kidney diseases.

[0003] In recent years, with the development of social economy and the improvement of living standards, people's dietary structure has gradually changed, resulting in a trend of younger age among HUA patients and a continuous increase in the number of patients. The significant characteristic of HUA patients is the abnormal elevation of blood uric acid concentration. High concentrations of uric acid accumulate and deposit in the joints, and then form urate crystals, which may induce joint inflammation and ultimately form gout.

[0004] In the clinical treatment of the acute phase of gout, we usually use non-steroidal anti-inflammatory drugs such as indomethacin and drugs such as colchicine to effectively relieve joint inflammation. Subsequently, to reduce the uric acid level, we use uric acid-lowering drugs, such as drugs that inhibit uric acid production, such as allopurinol and toperisone, or drugs that promote uric acid excretion, such as probenecid, sulfinpyrazone, and benzbromarone. These treatment measures aim to effectively control the condition and improve the quality of life of patients.

[0005] Although these drugs show certain efficacy in reducing uric acid, the accompanying side effects, such as allergic reactions, gastrointestinal discomfort, impaired liver and kidney functions, and an increased risk of cardiovascular and cerebrovascular diseases, cannot be ignored. These side effects greatly limit their wide application. Therefore, it is of great significance and practical application value to develop highly effective uric acid-lowering drugs with mild side effects and low toxicity for improving the treatment effect of patients and reducing the treatment risk. At the same time, developing a nutritional supplement that is both effective and safe and can reduce the blood uric acid level has an even broader market prospect. Summary of the Invention

[0006] The object of the present invention is to provide the use of β-alanine in the preparation of a drug for preventing and / or treating diseases related to uric acid metabolism disorders.

[0007] β-alanine, as a non-essential amino acid, participates in the synthesis of carnosine together with histidine in human physiological processes. Carnosine, mainly stored in skeletal muscle, has significant physiological effects, including but not limited to reducing the accumulation of lactic acid during exercise, neutralizing free radicals, alleviating oxidative stress, increasing nitric oxide levels, delaying the aging process, and improving cardiovascular health. However, due to the relatively high content of histidine and relatively low content of β-alanine in muscle, this to some extent limits the production of carnosine.

[0008] In recent years, in-depth exploration in the scientific research field has revealed the important value of β-alanine supplements. Research shows that by supplementing β-alanine, the level of carnosine in muscle can be significantly increased, up to 80%, thereby significantly enhancing muscle endurance, reducing acidosis, and improving the anaerobic exercise ability and strength of the human body. Given its safe and non-toxic properties, β-alanine, as a nutritional supplement, has been widely used among athletes and fitness enthusiasts.

[0009] On the one hand, based on rigorous scientific research, the present invention reveals the theoretical basis for β-alanine to reduce uric acid levels. In previous work, the applicant of the present invention constructed a hyperuricemia prediction model using non-target metabolome and targeted metabolomics data of hyperuricemia patients ( Figure 1 ), and through Rmcorr correlation analysis, it was found that a higher level of β-alanine and a lower level of uric acid had a strong correlation ( Figure 2 ).

[0010] On the other hand, through the intervention experiment of β-alanine on hyperuricemia mice, the present invention fully confirms that β-alanine can effectively reduce the blood uric acid level of hyperuricemic mice and promote urinary uric acid excretion. This discovery provides new possibilities for the prevention and / or treatment of diseases related to uric acid metabolism disorders.

[0011] Therefore, the present invention provides the use of β-alanine in the preparation of a drug for the prevention and / or treatment of diseases related to uric acid metabolism disorders.

[0012] Preferably, the drug is a drug having the effect of negatively regulating uric acid synthesis and / or promoting uric acid excretion in a hyperuricemia model.

[0013] Preferably, the effective concentration of the drug is about 3% by mass percentage.

[0014] The present invention also provides a drug for the prevention and / or treatment of diseases related to uric acid metabolism disorders, which contains β-alanine as an active ingredient.

[0015] Preferably, the drug further contains a pharmaceutically acceptable carrier composition.

[0016] Preferably, the pharmaceutical dosage form is a pharmaceutically acceptable oral preparation, and the dosage form is selected from: dry powder, granule, suspension, tablet or other suitable dosage forms.

[0017] The drug described above can be used for diseases related to uric acid metabolism disorders, including but not limited to one or more of hyperuricemia and its complications, gouty arthritis, or secondary kidney injury caused by hyperuricemia.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] 1. β-alanine, as the core component of the present invention, is a non-essential amino acid, has the function of nutritional supplement, and at the same time shows the characteristics of safety and non-toxicity.

[0020] 2. Compared with traditional chemical drugs for reducing uric acid, the drug of the present invention is friendly to the ecological environment, has no toxic side effects and residual risks, and shows the characteristics of green safety.

[0021] 3. The drug of the present invention is convenient and safe to use, has been applied in the human body as other efficacy products, has no potential hazards, and combines safety and high efficiency.

[0022] 4. The β-alanine of the present invention is an amino acid with uric acid-lowering effect screened from human hyperuricemia samples, specifically manifested in:

[0023] (1) There is a strong correlation with the reduction of uric acid;

[0024] (2) It can significantly reduce the uric acid level in hyperuricemic mice;

[0025] (3) Promote the urinary uric acid excretion in hyperuricemic mice.

[0026] Therefore, β-alanine shows great application potential and economic value in hyperuricemia, gout and other diseases caused by hyperuricemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 is the modeling flowchart of the hyperuricemia prediction model.

[0028] Figure 2 is the correlation between β-alanine and uric acid level.

[0029] Figure 3 is the effect of β-alanine on the body weight of hyperuricemic mice.

[0030] Figure 4 is the effect of β-alanine on the blood uric acid of hyperuricemic mice.

[0031] Figure 5 is the effect of β-alanine on the urinary uric acid of hyperuricemic mice.

[0032] Figure 6 Effect of β-alanine on serum and liver XOD in hyperuricemic mice.

[0033] Figure 7 Effect of β-alanine on kidney pathology and urate-excreting proteins in hyperuricemic mice. Detailed implementation manners

[0034] Hereinafter, we will elaborate on the present invention in detail and clearly based on the specific implementation technical solutions and the obtained experimental results. It should be clear that the described specific embodiments are only exemplary in nature, aiming to explain the principles and characteristics of the present invention, rather than limiting the scope of application of the present invention. In the test methods involved in the embodiments, unless otherwise specified, they all follow the conventional experimental methods; at the same time, for the experimental materials, reagents, etc. used, unless otherwise specified, they are all reagents and materials that can be obtained through commercial channels.

[0035] Example 1

[0036] (1) In-depth exploration of serum metabolites related to uric acid levels

[0037] 1. Sample selection: A total of 143 volunteers were recruited in this study and divided into three cohorts for in-depth research:

[0038] (1) Discovery cohort (n = 49): Composed of Han Chinese volunteers, with a male-to-female ratio of 26 / 23 and an average age of 35.08 ± 6.48 years. The volunteers in this cohort were transferred from the plain area to the plateau (Nyingchi City, altitude 2900 m) and stayed for 12 months. Clinical data and blood samples were collected at baseline, and repeated collections were performed at the 0th month (the 1st week), 1st, 3rd, 6th, and 12th months after high-altitude exposure.

[0039] (2) Validation cohort (n = 47): Also composed of Han Chinese volunteers, with a male-to-female ratio of 35 / 12 and an average age of 40.37 ± 6.06 years. These volunteers migrated from the plain area to the plateau and lived there for 12 months. Clinical data and blood samples were collected at baseline, and repeated collections were performed at the 6th and 12th months after high-altitude exposure.

[0040] (3) Local cohort (n = 47): This cohort was divided into two subgroups, including 36 people in the Han Chinese group (Local_H group) (male-to-female ratio = 10 / 26), with an average age of 30.86 ± 8.30 years; and 11 people in the Tibetan group (Local_T group) (male-to-female ratio = 1 / 10), with an average age of 35.73 ± 8.57 years. All were residents who had lived on the plateau for a long time, with a residence duration of more than five years. Clinical data and blood samples were collected from the volunteers in these two subgroups.

[0041] During the follow-up period, some Han Chinese volunteer Tibet aid workers left Nyingchi due to work arrangements and were thus considered to have withdrawn from this study. All blood samples were collected in a fasting state, and the serum was separated by centrifugation and stored at -80 °C for subsequent testing. According to whether the baseline serum uric acid level was higher than the normal upper limit of detection, the volunteers in each cohort were divided into a control group and a hyperuricemia group. The untargeted metabolomics analysis was performed on 259 serum samples from the discovery cohort and 47 serum samples from the validation cohort using the UHPLC-MS method. Meanwhile, the targeted detection of AAs, SCFAs, and MCFAs was performed on 426 serum samples from the discovery cohort, validation cohort, and local cohort using the UHPLC-MS / MS and GC-MS methods (as Figure 1 shown).

[0042] 2. Analysis strategy: In this study, a 10-fold cross-validation random forest classification (RFC) based on selected metabolites and phenotypes was used to construct a prediction model in the discovery cohort to identify serum metabolites related to serum uric acid levels under high-altitude exposure and to perform independent validation in the validation cohort. The local cohort was used to explore the effects of ethnicity and high-altitude exposure time on serum uric acid and metabolome. Through the Rmcorr correlation analysis, the association between metabolites and serum uric acid was deeply analyzed. To ensure the robustness of the research results, a sensitivity analysis was performed in the discovery cohort. In addition, after adjusting for age, gender, and BMI, a multivariable logistic regression model was used to further analyze the correlation between key serum metabolites and hyperuricemia levels.

[0043] (2) Pharmacodynamic experiment to verify the uric acid-lowering effect of β-alanine

[0044] 1. Experimental materials

[0045] The materials used in the experiment included 60 SPF-grade Kunming mice, all male, 4 weeks old, and weighing between 18 and 20 grams. The mice were purchased from the Guangdong Medical Experimental Animal Center and had a legal license number: Yue Si Zheng (2024) 05073. The experimental environment was strictly controlled at a temperature of 23 ± 2 °C and a humidity of 60 ± 10%, ensuring that the mice had free access to food and water. The reagents required for the experiment included β-alanine, potassium oxonate, hypoxanthine, allopurinol, etc., all purchased from Macklin Reagent Company; the uric acid assay kit was provided by Beijing Boxshengong Technology Co., Ltd.; the mouse xanthine oxidase (XOD) enzyme-linked immunosorbent assay kit was from Shanghai Future Industry Co., Ltd., and the primers were synthesized by Shanghai Shengong Biological Engineering Co., Ltd.; the real-time fluorescence quantitative PCR kit was from Nanjing Novozymes Biotechnology Co., Ltd. All experimental operations were carried out strictly according to the instructions of the kits.

[0046] 2. Experimental method

[0047] The experimental mice were randomly divided into six groups, namely the control group, the model group (PO+HX), the low-dose β-alanine administration group (1%), the medium-dose β-alanine administration group (2%), the high-dose β-alanine administration group (3%), and the allopurinol administration group (5mg / kg-ALL), with 10 mice in each group.

[0048] The mice in the control group were fed with regular food without any other treatment. While the mice in the model group and other administration groups were established with a hyperuricemia model by gavage with 900mg / kg potassium oxonate and 600mg / kg hypoxanthine daily for 14 consecutive days. During this period, the changes in the physiological states of the mice, such as body weight, hair color, diet, and drinking water, were recorded in detail.

[0049] During the administration process, the mice in the β-alanine administration groups were administered by freely drinking drinking water containing different concentrations (1%, 2%, 3%) of β-alanine. The mice in the allopurinol group were gavaged with 5mg / kg of allopurinol 1 hour after the modeling and administration, while the mice in the control group and the model group were given the same volume of solvent.

[0050] After 14 consecutive days of administration, the blood and urine specimens of the mice in each group were collected, and their uric acid levels were measured using a biochemical analyzer. Subsequently, the mice were anesthetized with sodium pentobarbital, and the liver and kidney tissues were isolated, weighed, and recorded. The right kidney was fixed in 4% paraformaldehyde to prepare 4μm paraffin sections for pathological examination with H&E staining. The remaining tissues were stored in liquid nitrogen for subsequent tests.

[0051] (III) Experimental Results and Discussion

[0052] 1. Through Rmcorr correlation analysis, we deeply explored the potential relationship between the serum uric acid level and serum metabolites in individuals exposed to high altitudes. The analysis results showed that there was a significant negative correlation between β-alanine and serum uric acid ( Figure 2 A). It is worth noting that compared with the residents in plain areas, the serum uric acid level of the permanent residents in high altitudes was significantly lower, while β-alanine showed an increasing trend in their bodies ( Figure 2 B), which may reveal the potential mechanism of action of β-alanine in reducing the serum uric acid level of residents with long-term high-altitude exposure.

[0053] 2. General state and body weight of mice

[0054] As Figure 3 shown, compared with the control group, the body weight of the animals in the modeling group increased slowly with the increase of modeling time. Especially, the difference in body weight between the modeling group and the normal group was significant after 14 days (P<0.05). Compared with the model group, the rising speed of the body weight of the animals in the allopurinol administration group and the β-alanine administration group was relatively slow ( Figure 3 ), but there were no obvious differences in appearance signs, behavioral activities, fecal traits, etc.

[0055] 3. β-Alanine reduces the serum uric acid level in a hyperuricemia mouse model induced by potassium oxonate and hypoxanthine

[0056] As Figure 4 shown, the serum uric acid content of the mice in the model group increased significantly, showing a significant difference compared with the normal group (P < 0.0001), indicating successful modeling. After continuous daily administration of allopurinol and low, medium, and high doses of β-alanine for 14 days, the serum uric acid content of the mice in the administration groups decreased significantly, and showed a downward trend with the increase in the concentration of β-alanine: the medium-dose β-alanine administration group (2%) had a significant uric acid-lowering effect compared with the model group (P < 0.001), and the high-dose β-alanine administration group (3%) had the best uric acid-lowering effect similar to the allopurinol administration group. The experimental results prove that the β-alanine of the present application has the efficacy of reducing the uric acid level.

[0057] 4. β-Alanine promotes the urinary uric acid excretion in a hyperuricemia mouse model induced by potassium oxonate and hypoxanthine

[0058] As Figure 5 shown, after modeling, the urinary uric acid level of the mice in the model group increased significantly, showing a significant difference compared with the normal group (P < 0.0001), indicating successful modeling. The urinary uric acid increased in the low-dose β-alanine group, which was significantly higher than that of the normal control group, indicating increased uric acid excretion. Allopurinol is an isomer of hypoxanthine, and it and its metabolites reduce uric acid production and lower the serum uric acid and urinary uric acid concentrations by inhibiting XOD. After continuous daily administration of allopurinol and different doses of β-alanine for 14 days, the urinary uric acid content of the mice in the high-dose β-alanine administration group decreased significantly, showing no significant difference from the allopurinol group, indicating that the reduction in uric acid production secondary to the reduction in urinary uric acid excretion.

[0059] 5. Effects of β-alanine on serum and liver XOD in hyperuricemia mice

[0060] As Figure 6 shown, after modeling, the serum and liver XOD contents of the mice in the model group increased, showing a significant difference compared with the normal group (P < 0.0001). After continuous daily administration of allopurinol and low, medium, and high doses of β-alanine for 14 days, the serum and liver XOD contents of the mice in the administration groups decreased, and the down-regulation of XOD in the liver was more obvious (P < 0.01). The urinary uric acid content of the mice in the high-dose β-alanine administration group decreased significantly, showing no significant difference from the allopurinol group. It can be seen that β-alanine reduces uric acid production and lowers the serum uric acid and urinary uric acid concentrations by reducing liver XOD.

[0061] 6. Effects of β-alanine on kidney pathology and uric acid excretion-promoting proteins in hyperuricemic mice.

[0062] As shown Figure 7 in Figure 7 Figure A, compared with the β-alanine intervention group, the pathological manifestations of the kidneys in the model group were thickening of the basement membrane, sclerosis of some glomeruli, and thickening and hyaline degeneration of the renal arterioles Figure 7 (Figure A). The expression of ABCG2, which promotes the excretion of uric acid, was increased in the β-alanine group, higher than that in the model group

[0063] (Figure B). On the contrary, the expressions of GLUT9 and URAT1, which mediate the reabsorption of uric acid, were lower in the β-alanine group than in the model group. It can be speculated from this that β-alanine intervention may enhance the excretion of uric acid in hyperuricemic mice.

[0064] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. It is not necessary and impossible to enumerate all the implementation manners here. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of β-alanine in the preparation of a medicament for reducing the serum uric acid concentration level.

2. The application according to claim 1, wherein The medicament is used for preventing and / or treating diseases related to uric acid metabolism disorders, wherein the diseases related to uric acid metabolism disorders include, but are not limited to, one or more of hyperuricemia and its complications, gouty arthritis, or secondary renal injury caused by hyperuricemia.

3. The application according to claim 1, characterized in that, The medicament is a medicament having the effect of negatively regulating uric acid synthesis in hyperuricemia and / or promoting uric acid excretion.

4. The application according to claim 1, characterized in that The effective concentration of β-alanine in the medicament is about 3% by mass percentage.

5. The application according to claim 1, wherein The medicament further comprises at least one pharmaceutically acceptable carrier.

6. The application according to claim 1, characterized in that The dosage form of the medicament is a pharmaceutically acceptable oral preparation, and the dosage form is selected from: dry powder, granule, suspension, tablet or other suitable dosage forms.

7. A pharmaceutical composition for preventing and / or treating diseases related to uric acid metabolism disorders, characterized in that, It contains an effective therapeutic amount of β-alanine as an active ingredient, and at least one pharmaceutical excipient selected from disintegrants, binders and optionally antioxidant additives as a pharmaceutical carrier.

8. The pharmaceutical composition according to claim 7, wherein The dosage of the β-alanine is 3% of the daily water intake, and it is uniformly administered orally.

9. The pharmaceutical composition according to claim 7, characterized in that, The dosage form is selected from one or more of dry powder, granule, suspension, tablet or other suitable dosage forms.

10. The pharmaceutical composition according to claim 7, characterized in that, The diseases related to uric acid metabolism disorders include, but are not limited to, one or more of hyperuricemia and its complications, gouty arthritis, or secondary renal injury caused by hyperuricemia.

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