Application of undaria pinnatifida sporophyll fucoidan in preparation of products for reducing uric acid or treating gout

Abstract

The invention relates to novel application of undaria pinnatifida sporophyll fucoidin, and belongs to the technical field of food and medicine. The invention provides a new application of undaria pinnatifida sporophyll fucoidan in preparation of drugs, health food and food for reducing uric acid, and the undaria pinnatifida sporophyll fucoidan can remarkably reduce the blood uric acid level and relieve and treat gout symptoms, is free of toxic and side effects, safe and effective, can be used for preparing URAT1 inhibitors and / or GLUT9 inhibitors, and has a good application prospect. The compound can be used for preparing medicines or health foods and foods for reducing uric acid and treating gout, and has a wide application prospect.

Description

Technical Field

[0001] The invention relates to a new application of fucoidan from spore leaves of Undaria pinnatifida, and belongs to the technical field of food and medicine. Background Art

[0002] The sporophylls of Undaria pinnatifida (Mekabu) are the parts of Undaria pinnatifida where spores are produced. They are located between the leaves and the rhizoids of Undaria pinnatifida and are rich in fucoidan. The chemical structure of fucoidan is very complex, and the structures of fucoidan isolated from different brown algae vary greatly. The basic structure of fucoidan (MF) from the sporophylls of Undaria pinnatifida is composed of sulfated fucose and galactose, with a molar ratio of about 1:1, and some are accompanied by acetylation, such as Figure 1 As shown. The fucoidan of Laminaria saccharina is composed of a poly-α-(1→3)-furanofucoside backbone, with the sulfate group mainly located at the C-4 position, sometimes at the C-2 position, and acetylated at the C-2 position in some cases; the fucoidan of Cladosiphonokamuranus also has an α(1→3)-linked L-furanofucose backbone, which can be substituted with sulfate at the C-4 position and / or through a single α(1→2)-linked α-L-furanofucose substituent, accompanied by adjacent glucuronic acid substitutions, and some side chain fucose residues may be O-acetylated, which is significantly different from the MF molecular formula, as shown in Figure 2. Figure 2 As shown in the figure. Fucoidan from different sources has significant differences in electron microscope microstructure. MF is significantly different from fucoidan from other sources, which may be due to its different molecular weight or main chain conformation, which affects the different interactions within the polysaccharide molecules and forms different structures, thus producing diverse biological activities. MF has multiple activities, such as antioxidant, anti-tumor, anticoagulant, immunomodulatory, antibacterial and antiviral, hypoglycemic and hypolipidemic, and neuroprotective.

[0003] Uric acid is one of the metabolites of the human body. It is mainly produced by the liver metabolism of purine compounds ingested by the diet and decomposed in the body, and excreted through the kidneys and digestive tract. Under normal circumstances, the production and excretion of uric acid in the body are in balance. Hyperuricemia (HUA) is a metabolic disease caused by purine metabolism disorders and / or abnormal uric acid excretion. Its main clinical feature is increased blood uric acid in the body. The kidney is an important organ for uric acid excretion. About 2 / 3 of uric acid is excreted from the body through the kidneys with urine, and a small part is excreted through the gastrointestinal tract. Some HUA patients will eventually develop gout or other chronic diseases, which seriously affect the quality of life of patients. HUA is not only the biochemical basis of gout, but also an independent risk factor for the development of chronic kidney disease, cardiovascular disease and various metabolic diseases, and an independent predictor of all-cause mortality. However, it is often ignored because of its hidden clinical symptoms.

[0004] At present, the prevention and treatment of HUA mainly rely on a low-purine diet and uric acid-lowering drugs. However, exogenous purines only account for about 20% of the total purines in the human body. Long-term excessive low-purine diets not only have limited effects on the prevention and treatment of HUA, but may also lead to nutritional deficiencies. Although the existing uric acid-lowering drugs have clear pharmacological mechanisms and obvious uric acid-lowering effects, long-term use can cause serious liver and kidney damage. Drugs used clinically to lower uric acid levels mainly include: (1) Drugs that inhibit uric acid production: xanthine oxidase inhibitors (such as allopurinol and febuxostat), which reduce uric acid production by inhibiting xanthine oxidase (XO); however, these drugs may cause side effects such as fever, diarrhea, allergic reactions, and liver damage, and some patients have poor tolerance; (2) Drugs that promote uric acid excretion: uric acid transporter inhibitors (such as benzbromarone and probenecid), which inhibit the expression of the renal uric acid reabsorption transporter URAT1. , reducing the reabsorption of uric acid by the renal tubules, accelerating the renal excretion of uric acid, and lowering the level of uric acid in the blood; however, this type of drug may cause side effects such as fever, diarrhea, allergic reactions, and liver damage, and some patients have poor tolerance; (3) Uricase preparations: recombinant uricase such as rebuxostat, which degrades uric acid into more soluble products, 49% of which is excreted through the kidneys and 45% through the feces. It is a dual-channel excretion drug that promotes excretion; however, it may cause harm to cardiovascular health, the drug has limited clinical application, is expensive, and is not suitable for long-term use.

[0005] Due to the limitations of existing drugs, there is an urgent need to develop new, safe, effective, and uniquely structured drugs, health foods, and foods for preventing, treating, and alleviating HUA and diseases caused by hyperuricemia (gout, etc.). Summary of the invention

[0006] The purpose of the present invention is to provide new applications of MF in medicines, health foods, and foods for preventing, treating, and alleviating HUA and diseases caused by hyperuricemia (gout, etc.), overcome the shortcomings of the prior art, and provide a safe, effective, and unique mechanism of action natural uric acid-lowering solution.

[0007] In order to achieve the above-mentioned invention object, the present invention adopts the following technical solutions:

[0008] The invention provides application of Undaria pinnatifida spore leaf fucoidan (Mekabu Fucoidan, MF) in preparing medicines, health foods and foods for reducing uric acid.

[0009] In the above technical scheme, further, the application of the spore leaf fucoidan of Undaria pinnatifida in the preparation of medicines for preventing, treating and alleviating diseases related to hyperuricemia; the application of the spore leaf fucoidan of Undaria pinnatifida in the preparation of health foods and foods for preventing and alleviating diseases related to hyperuricemia.

[0010] In the above technical solution, further, the hyperuricemia-related disease is gout caused by hyperuricemia.

[0011] In the above technical solution, further, the active ingredient in the food, health food or medicine includes Undaria pinnatifida spore leaf fucoidan.

[0012] In the above technical solution, further, the effective dose of the Undaria pinnatifida spore leaf fucoidan is ≥450 mg / day.

[0013] In the above technical solution, further, the effective dosage of the Undaria pinnatifida spore leaf fucoidan is 450-1350 mg / day.

[0014] In the above technical solution, further, the spore leaf fucoidan of Undaria pinnatifida significantly reduces the reabsorption of uric acid and increases uric acid excretion by inhibiting the renal uric acid transporters URAT1 and GLUT9; the spore leaf fucoidan of Undaria pinnatifida has a certain inhibitory effect on the liver xanthine oxidase (XO), playing an auxiliary role in lowering uric acid. The spore leaf fucoidan of Undaria pinnatifida has antioxidant and anti-inflammatory properties, which indirectly promotes uric acid metabolism and excretion.

[0015] In the above technical solution, further, the basic structure of the Undaria pinnatifida spore leaf fucoidan is as follows, which is mainly composed of sulfated fucose and galactose, with a molar ratio of about 1:1, and some are accompanied by acetylation:

[0016]

[0017] In the above technical solution, further, the dosage form of the product includes oral liquid, capsule, tablet, granule or targeted agent.

[0018] The present invention also provides the use of Undaria pinnatifida spore leaf fucoidan in the preparation of URAT1 inhibitor and / or GLUT9 inhibitor.

[0019] In the above technical solution, further, the active ingredients of the URAT1 inhibitor and the GLUT9 inhibitor include fucoidan from the spore leaves of Undaria pinnatifida.

[0020] In the above technical solution, further, the effective dose of the fucoidan from the spore leaves of Undaria pinnatifida is ≥450 mg / day; preferably, the effective dose of the fucoidan from the spore leaves of Undaria pinnatifida is 450-1350 mg / day.

[0021] Beneficial effects of the present invention:

[0022] The present invention proposes for the first time that the spore leaf fucoidan MF of Undaria pinnatifida has the effect of lowering uric acid, is safe and non-toxic, and can effectively alleviate the frequency and severity of gout attacks. The spore leaf fucoidan of Undaria pinnatifida mainly inhibits URAT1 and GLUT9 through its high binding force with URAT1 and GLUT9, significantly reducing the reabsorption of uric acid in the renal tubules and increasing the excretion of uric acid, thereby reducing serum uric acid levels. MF can be used to prepare URAT1 inhibitors and GLUT9 inhibitors to lower uric acid. MF is safe to take, can effectively lower uric acid, and can be used to prepare drugs or health foods and foods for lowering uric acid and treating gout, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 MF basic structure;

[0024] Figure 2 The basic structure of fucoidan from kelp and Okinawa mozuku;

[0025] Figure 3 Changes in feed intake in mice after a single dose of MF toxicity test;

[0026] Figure 4 Body weight changes of mice in the toxicity test of MF single administration;

[0027] Figure 5 Changes in organ coefficients of mice in the toxicity test of MF single administration;

[0028] Figure 6 Changes in uric acid in 20 subjects before and after taking MF;

[0029] Figure 7 Patient A's uric acid change curve before and after taking MF from 2016 to 2024;

[0030] Figure 8 Uric acid change curve of patient B before and after taking febuxostat and MF. DETAILED DESCRIPTION

[0031] The present invention is further described below in conjunction with specific embodiments, but the present invention is not limited in any way.

[0032] Example 1

[0033] Toxicology experiments of MF:

[0034] 1. Experimental samples: The MF components used in this toxicological experiment are: fucose 21.0%, galactose 20.6%, sulfate content 25.3%, number average molecular weight 95,000, weight average molecular weight 437,000. The molecular weight distribution of MF is shown in Table 1.

[0035] Table 1

[0036]

[0037] 2. Acute oral toxicity test in mice

[0038] Experimental method: The acute toxicity of MF was detected by acute oral toxicity test in mice.

[0039] Experimental subjects: 20 male and 20 female Kunming mice, weighing 17.5-22.9 g.

[0040] Dose setting: A single oral administration of 4000 mg / kg body weight of MF was used as the maximum tolerated dose (MTD) experiment.

[0041] Observation period: 14 days.

[0042] 3. Experimental results:

[0043] Survival status: During the observation period, no death or poisoning symptoms were found in the experimental mice.

[0044] General status: There are no abnormalities in the activities, diet, drinking water, fur, feces, etc. of male and female animals. Figure 3 The figure shows the living conditions (diet) of male and female mice. The feed intake of the treated group was slightly lower than that of the control group, but the difference was not statistically significant, so there was no abnormality in the diet after taking MF.

[0045] Weight changes: The weight gain of mice in the experimental group and the control group was normal, with no significant difference ( Figure 4 ). Organ coefficient: The organ coefficients of the main organs (heart, liver, spleen, lung, kidney, pancreas) were not significantly different from those of the control group ( Figure 5 ) Autopsy results: After the observation period, the mice were autopsied and no pathological changes were found in their internal organs and tissues.

[0046] Under the current experimental conditions, MF has an effect on the LD of mice. 50 The values ​​(median lethal dose) are all greater than 4000 mg / kg body weight, and MF is a practically non-toxic substance.

[0047] 4. Safety evaluation

[0048] According to the toxicology evaluation standard: According to the results of acute toxicity experiments, MF has no acute toxicity, which proves that MF has extremely high safety and can be used in the development of drugs, health foods and foods.

[0049] Example 2

[0050] In the course of evolution, the gene encoding urate oxidase (UOX) in humans has been silenced and inactivated, so the risk of HUA is greatly increased. However, the gene encoding urate oxidase in rodents (mice) is normally expressed, and urate oxidase can break down uric acid into allantoin and excrete it from the body. Therefore, it is difficult for the mouse model to simulate the HUA model, and its stability and accuracy are limited. Therefore, this experiment widely recruited volunteers from the population.

[0051] 1. Test sample: The MF sample used this time is consistent with the toxicological test sample in Example 1.

[0052] 2. Population trial plan

[0053] People with blood uric acid levels greater than 420 μmol / L (some of whom have developed gout) are tested as follows:

[0054] Experimental method: Do not take other uric acid-lowering drugs, take MF twice a day, on an empty stomach, once in the morning and once in the evening, 450 mg each time, for 30 consecutive days.

[0055] Subjects: 20 subjects, age range 30-60 years old.

[0056] Test results ( Figure 6 ):

[0057] Reduced uric acid levels: 85% of the subjects had reduced blood uric acid levels: 55% of the subjects had uric acid levels reduced by more than 50μmol / L, and 20% of the subjects had uric acid levels reduced by more than 100μmol / L.

[0058] Improvement in gout symptoms: Subjects experienced a significant reduction in the frequency and severity of gout attacks.

[0059] Safety observation: During the trial, no adverse reactions related to MF were found.

[0060] 3. Population test cases

[0061] Case 1: A case study of long-term quantitative use of MF, as follows:

[0062] Male patient A, born in 1953, is a HUA patient.

[0063] Medical history: The patient began monitoring his uric acid level in 2016, and the uric acid value remained high.

[0064] 2016: 440 μmol / L,

[0065] 2017: 437 μmol / L,

[0066] 2018: 452 μmol / L,

[0067] 2019: 485 μmol / L,

[0068] Interventions: Starting in early 2020, patients took MF 450 mg daily.

[0069] Uric acid monitoring results ( Figure 7 ):

[0070] 2020 (after taking it for half a year): 386μmol / L,

[0071] 2021: 398 μmol / L,

[0072] 2022: 394 μmol / L,

[0073] 2024: 284 μmol / L,

[0074] Result analysis:

[0075] Uric acid levels decreased significantly: uric acid levels in 2024 decreased by 201 μmol / L compared with 2019.

[0076] Long-term stability: Uric acid levels remain stable and gradually decrease during the period of taking MF.

[0077] Safety and Tolerability: Patients did not report any adverse reactions or symptoms.

[0078] Case 2: A case study comparing the effects of taking MF and medication, as follows:

[0079] Medical history: Male patient B, born in 1966, HUA and gout patient, gout for more than 10 years, uric acid level was 520μmol / L when not taking medication. The patient took 20mg / day of febuxostat all year round, and uric acid was maintained at 360μmol / L.

[0080] Intervention measures: To compare the effects of taking MF, the patient stopped taking febuxostat for 7 days, and the uric acid level returned to 520 μmol / L. After that, MF was taken once a day, on an empty stomach, 450 mg each time, for 15 consecutive days. The changes in uric acid levels are shown in Figure 8 .

[0081] Uric acid monitoring results: The uric acid level decreased significantly on the second day of taking it, and on the 13th day of taking it, the uric acid level had dropped to 380μmol / L, which was lower than the critical value (420μmol / L). After 15 days of stopping MF, the uric acid level rose to 520μmol / L. The patient did not report any adverse reactions or discomfort symptoms. The data showed that the effect of MF on lowering uric acid levels was comparable to that of Febuxostat.

[0082] Case 3-12: The gender, age, dosage, duration of use, and uric acid levels before and after use of Case 3-12 are detailed in Table 2. Among them, 4 patients took 450 mg three times a day, and their uric acid levels dropped by 120-340 μmol / L; 5 patients took 450 mg twice a day, and their uric acid levels dropped by 60-139 μmol / L; 1 patient took 450 mg once a day, and their uric acid levels dropped by 30 μmol / L.

[0083] Table 2 Changes of uric acid in 10 subjects before and after taking MF

[0084]

[0085] The above 10 subjects took MF and their uric acid levels were significantly reduced, and no adverse reactions or discomfort symptoms were reported, which proves its uric acid-lowering effect and safety in long-term use, and can be used as an effective means to treat HUA and gout.

[0086] Example 3

[0087] Mechanism of action of MF in lowering uric acid and treating gout:

[0088] Using artificial intelligence technology, molecular docking, molecular dynamics simulation, enhanced sampling technology and quantum mechanics / molecular mechanics (QM / MM) simulation, the mechanism of MF in lowering uric acid was studied, mainly including:

[0089] 2.1 Inhibit the function of renal uric acid transporter (URAT1) in the kidney

[0090] High binding affinity: After simulation, the binding free energy of MF to URAT1 is about -40 kcal / mol, which is higher than that of known URAT1 inhibitors (such as benzbromarone and probenecid). The binding free energy of MF to GLUT9 is about -35 kcal / mol, showing strong binding ability.

[0091] Key interactions: MF binds to key sites of these transporters (such as Arg477 and Lys35 of URAT1 and Arg380 and Lys394 of GLUT9), hindering the reabsorption of uric acid in the renal tubules. The fucose methyl substitution and L-configuration of MF enhance the hydrophobic and electrostatic interactions with URAT1, and the galactose hydroxyl groups of MF form hydrogen bonds with proteins, which together improve the binding stability.

[0092] Effect: Inhibition of URAT1 and GLUT9 can significantly reduce the reabsorption of uric acid in the renal tubules, increase the excretion of uric acid, and thus reduce serum uric acid levels.

[0093] 2.2 Assist in inhibiting the activity of xanthine oxidase (XO) in the liver

[0094] Binding affinity: The sulfate group of MF forms strong electrostatic interactions and hydrogen bonds with key residues in the active site of XO (such as Glu802 and Arg880), and the binding free energy with XO is about -30 kcal / mol, which is lower than that of known XO inhibitors (such as allopurinol and febuxostat).

[0095] Limited effect: The inhibitory effect on XO is weak and may only partially reduce the production of uric acid, playing a role in assisting in lowering uric acid.

[0096] 2.3 Effects on other uric acid transporters

[0097] Low binding affinity: The binding free energy of MF to OAT1, OAT3, ABCG2 and other proteins is around -25 kcal / mol, and the effect is relatively weak.

[0098] Limited impact: No significant effect on the secretion and excretion of uric acid, and little effect on lowering uric acid.

[0099] 2.4 Reduce inflammatory response

[0100] The occurrence of HUA and gout is closely related to oxidative stress. Excessive accumulation of uric acid can lead to the production of free radicals, causing tissue damage. MF has a strong antioxidant capacity, can scavenge free radicals, and reduce the level of oxidative stress. MF indirectly promotes the metabolism and excretion of uric acid by reducing inflammation and oxidative stress and improving the tissue microenvironment.

Claims

1. Application of Undaria pinnatifida spore leaf fucoidan in the preparation of medicines, health foods and foods for lowering uric acid.

2. The use according to claim 1, characterized in that: The application of the Undaria pinnatifida spore leaf fucoidan in the preparation of medicines for preventing, treating and alleviating hyperuricemia and related diseases caused by it; the application of the Undaria pinnatifida spore leaf fucoidan in the preparation of health care foods and foods for preventing and alleviating hyperuricemia and related diseases caused by it.

3. The use according to claim 2, characterized in that: The hyperuricemia-related disease is gout caused by hyperuricemia.

4. The use according to claim 1, characterized in that: The active ingredient in the food, health food or medicine includes Undaria pinnatifida spore leaf fucoidan.

5. The use according to claim 1, characterized in that: The effective dosage of the Undaria pinnatifida spore leaf fucoidan is ≥450 mg / day.

6. The use according to claim 5, characterized in that: The effective dosage of the Undaria pinnatifida spore leaf fucoidan is 450-1350 mg / day.

7. The use according to claim 1, characterized in that: Undaria pinnatifida fucoidan can significantly reduce uric acid reabsorption and increase uric acid excretion by inhibiting renal uric acid transporters URAT1 and GLUT9; Undaria pinnatifida fucoidan has a certain inhibitory effect on liver xanthine oxidase (XO), playing an auxiliary role in lowering uric acid; Undaria pinnatifida fucoidan has antioxidant and anti-inflammatory properties, indirectly promoting uric acid metabolism and excretion.

8. The use according to claim 1, characterized in that: The basic structure of the spore leaf fucoidan of Undaria pinnatifida:

9. The use according to claim 1, characterized in that: The dosage forms of the product include oral solution, capsule, tablet, granule or targeted agent.

10. Use of fucoidan from spore leaves of Undaria pinnatifida in the preparation of URAT1 inhibitors and / or GLUT9 inhibitors.

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