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

Abstract

The present invention relates to a new application of fucoidan from sporophyll of Undaria pinnatifida, belonging to the technical field of food and medicine. The present invention provides a new application of fucoidan from sporophyll of Undaria pinnatifida in the preparation of drugs, health foods and foods for reducing uric acid, which can significantly reduce the level of blood uric acid, relieve and treat gout symptoms, has no toxic and side effects, is safe and effective, can be used for preparing URAT1 inhibitors and / or GLUT9 inhibitors, can be used for preparing health foods and foods for reducing uric acid, and can be used for preparing drugs for reducing uric acid and treating gout, and has broad application prospects.

Description

Technical Field

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

[0002] The sporophyll (Mekabu) of Undaria pinnatifida is the part where Undaria pinnatifida produces spores. It is located between the blade and the rhizoid of Undaria pinnatifida and is rich in fucoidan. Fucoidan has a very complex chemical structure, and the structures of fucoidans isolated from different brown algae vary greatly. The basic structure of fucoidan from the sporophyll of Undaria pinnatifida (Mekabu Fucoidan, abbreviated as MF) is composed of sulfated fucose and galactose, with a molar ratio of about 1:1, and individual ones are accompanied by acetylation, such as Figure 1 shown. The fucoidan of Laminaria saccharina is composed of a poly-α-(1→3)-furanfucoside main chain, and the sulfate groups are mainly located at the C-4 position, sometimes also at the C-2 position, and individual ones are acetylated at the C-2 position; the fucoidan of Cladosiphon okamuranus also has an α(1→3)-linked L-furanfucose main chain, which can be sulfated at the C-4 position and / or substituted by a single α-L-furanfucose substituent linked by α(1→2), and is accompanied by adjacent glucuronic acid substitution. Some fucose residues in the side chain may undergo O-acetylation, which is significantly different from the MF molecular formula, such as Figure 2 shown. Different sources of fucoidan have significant differences in the electron microscopy microstructure. There are significant differences between MF and fucoidans from other sources, which may be due to differences in their molecular weight or main chain conformation, thus affecting the different interactions within the polysaccharide molecule, forming different structures, and thus producing diverse biological activities. MF has various activities, such as antioxidant, anti-tumor, anticoagulant, immunomodulatory, antibacterial and antiviral, hypoglycemic and lipid-lowering, and neuroprotective.

[0003] Uric acid is one of the human body's metabolites, mainly produced by the liver metabolism of purine compounds ingested from the diet and decomposed in the body, and excreted through the kidneys and the digestive tract. Under normal circumstances, the production and excretion of uric acid in the body remain in a balanced state. Hyperuricemia (HUA) is a metabolic disease caused by purine metabolism disorders and / or abnormal uric acid excretion, and its main clinical feature is the elevation of blood uric acid in the body. The kidney is an important organ for uric acid excretion. Approximately two-thirds 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, seriously affecting the quality of life of patients. At present, the total prevalence of HUA in China has climbed to 18.4%, and the prevalence in young men (18 - 29 years old) reaches 32.3%, showing a significant trend of getting younger. HUA is not only the biochemical basis of gouty diseases, but also an independent risk factor for the occurrence and development of chronic kidney disease, cardiovascular disease and various metabolic diseases, as well as an independent predictor of all-cause mortality. However, it is often overlooked due to 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 diet not only has 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 will cause serious liver and kidney damage. Clinically used drugs for reducing uric acid levels mainly include: (1) Drugs that inhibit uric acid production: Xanthine oxidase inhibitors (such as allopurinol, 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 function damage, and some patients have poor tolerance; (2) Drugs that promote uric acid excretion: Urate transporter inhibitors (such as benzbromarone, probenecid), which reduce the reabsorption of uric acid in the renal tubules by inhibiting the expression of the renal uric acid reabsorption transporter URAT1, accelerate the renal excretion of uric acid, and reduce blood uric acid levels; but these drugs may cause side effects such as fever, diarrhea, allergic reactions, and liver function damage, and some patients have poor tolerance; (3) Uricase preparations: Recombinant uricases such as rasburicase, which degrade uric acid into more soluble products, 49% excreted through the kidneys and 45% excreted through the feces, belonging to dual-channel excretion drugs to promote excretion; but it may be harmful to cardiovascular health, with limited clinical application of the drug and high price, and is not suitable for long-term use.

[0005] Due to the limitations of existing drugs, there is an urgent need to develop new drugs that are safe, effective, and have unique mechanisms of action for the prevention, treatment, and alleviation of HUA and diseases caused by hyperuricemia (such as gout). Summary of the Invention

[0006] The object of the present invention is to provide a new application of Mekabu Fucoidan (MF) in the preparation of drugs for preventing, treating, and alleviating hyperuricemia (HUA) and diseases caused by hyperuricemia (such as gout, etc.), to overcome the deficiencies of the prior art, and to provide a safe, effective, and unique natural uric acid-lowering solution.

[0007] To achieve the above object of the invention, the following technical solutions are adopted:

[0008] The present invention provides the application of Mekabu Fucoidan (MF) in the preparation of drugs and foods for reducing uric acid.

[0009] In the above technical solution, further, the application of the Mekabu Fucoidan in the preparation of drugs for preventing, treating, and alleviating diseases related to hyperuricemia; the application of the Mekabu Fucoidan in the preparation of foods for alleviating diseases related to hyperuricemia.

[0010] In the above technical solution, further, the diseases related to hyperuricemia are gout caused by hyperuricemia.

[0011] In the above technical solution, further, the active ingredients in the food, health food, or drug include Mekabu Fucoidan.

[0012] In the above technical solution, further, the effective dose of the Mekabu Fucoidan ≥ 450 mg / day.

[0013] In the above technical solution, further, the effective dose of the Mekabu Fucoidan is 450 - 1350 mg / day.

[0014] In the above technical solution, further, the Mekabu Fucoidan significantly reduces the reabsorption of uric acid and increases uric acid excretion by inhibiting the renal uric acid transporters URAT1 and GLUT9; the Mekabu Fucoidan has a certain inhibitory effect on hepatic xanthine oxidase (XO), playing an auxiliary role in reducing uric acid. The Mekabu Fucoidan has antioxidant and anti-inflammatory properties, indirectly promoting uric acid metabolism and excretion.

[0015] In the above technical solution, further, the basic structure of the Mekabu Fucoidan is as follows, mainly composed of sulfated fucose and galactose, with a molar ratio of about 1:1, and individual acetylation:

[0016]

[0017] In the above technical solution, further, the dosage forms of the product include oral liquid, capsule, tablet, granule, or targeting agent.

[0018] The present invention also provides the use of laminaria japonica sporophyll fucoidan in the preparation of URAT1 inhibitors and / or GLUT9 inhibitors.

[0019] In the above technical solution, further, the active ingredients of the URAT1 inhibitor and the GLUT9 inhibitor include laminaria japonica sporophyll fucoidan.

[0020] In the above technical solution, further, the effective dose of the laminaria japonica sporophyll fucoidan ≥ 450 mg / day; preferably, the effective dose of the laminaria japonica sporophyll fucoidan is 450 - 1350 mg / day.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] The present invention first proposes that laminaria japonica sporophyll fucoidan MF has the effect of reducing uric acid, is safe and non-toxic, and can effectively relieve the attack frequency and severity of gout. Laminaria japonica sporophyll fucoidan mainly inhibits the reabsorption of uric acid in the renal tubules and increases the excretion of uric acid by significantly reducing URAT1 and GLUT9 through its high binding affinity with URAT1 and GLUT9, thereby reducing the serum uric acid level. MF can be used to prepare URAT1 inhibitors and GLUT9 inhibitors to reduce uric acid. MF is safe to take and can effectively reduce uric acid, and is used to prepare health foods and foods for reducing uric acid, as well as drugs for reducing uric acid and treating gout, and has broad application prospects. Description of the Drawings

[0023] Figure 1 Basic structure of MF;

[0024] Figure 2 Basic structures of fucoidans from Undaria pinnatifida and Gracilaria chorda;

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

[0026] Figure 4 Changes in the body weight of mice in the single-dose toxicity test of MF;

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

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

[0029] Figure 7 Uric acid change curve of patient A 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 implementation manners

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

[0032] Embodiment 1

[0033] Toxicology experiment of MF:

[0034] 1. Experimental sample: The MF components used in this toxicology experiment: fucose 21.0%, galactose 20.6%, sulfate group 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 for details.

[0035] Table 1

[0036] 2. Acute oral toxicity experiment of mice

[0037] Experimental method: The acute oral toxicity experiment method of mice was adopted to detect the acute toxicity of MF.

[0038] Experimental subjects: 20 male and 20 female Kunming mice, with a body weight of 17.5 - 22.9 g.

[0039] Dose setting: MF at a dose of 4000 mg / kg body weight was administered orally once for the maximum tolerated dose (MTD) experiment.

[0040] Observation period: 14 days.

[0041] 3. Experimental results:

[0042] Survival situation: During the observation period, no deaths or poisoning symptoms were found in the experimental mice.

[0043] General state: The living states of male and female animals such as activity, diet, drinking water, hair, feces, etc. were all normal. As Figure 3 shown, for the living states (diet) of male and female mice, the feed intake of the treatment group was slightly lower than that of the control group, but the difference was not statistically significant compared with the control group, so there was no abnormality in diet after taking MF.

[0044] Body weight change: The body weights of the mice in the experimental group and the control group increased normally, with no significant difference ( Figure 4 ) Organ coefficients: The organ coefficients of the main viscera (heart, liver, spleen, lung, kidney, pancreas) had no significant difference compared with the control group ( Figure 5 ). Anatomical results: After dissection at the end of the observation period, there were no pathological changes in the internal organs and tissues of the mice.

[0045] Under the current experimental conditions, the LD of MF for mice 50The values (median lethal dose) are all greater than 4000 mg / kg body weight, and MF belongs to substances with virtually no toxicity.

[0046] 4. Safety evaluation:

[0047] According to the toxicological evaluation criteria: Based on the results of the acute toxicity experiment, MF has no acute toxic effect. It is proved that MF has extremely high safety and can be used for the development of drugs, health foods, and foods.

[0048] Example 2

[0049] During human evolution, the gene encoding urate oxidase (UOX) was silenced and inactivated, so the risk of developing HUA has also increased significantly. In contrast, in rodents (mice), the gene encoding urate oxidase is normally expressed, and urate oxidase can decompose 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 trial widely recruited human volunteers.

[0050] 1. Test sample: The MF sample used in this trial is the same as the sample used in the toxicological experiment of Example 1.

[0051] 2. Human trial protocol

[0052] People with a blood uric acid level greater than 420 μmol / L (some have developed gout) were tested as follows:

[0053] Test method: Without taking other drugs for reducing blood uric acid, 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.

[0054] Subjects: 20 subjects, aged 30 - 60 years.

[0055] Test results ( Figure 6 )

[0056] Reduction in uric acid level: The blood uric acid levels of 85% of the subjects decreased: Among them, the uric acid levels of 55% of the subjects decreased by more than 50 μmol / L, and the uric acid levels of 20% of the subjects decreased by more than 100 μmol / L.

[0057] Improvement in gout symptoms: The frequency and severity of gout attacks in the subjects decreased significantly.

[0058] Safety observation: During the trial, no adverse reactions related to MF were found.

[0059] 3. Individual cases in the human trial

[0060] Case 1: Case study of long-term quantitative administration of MF is as follows:

[0061] Male patient A, born in 1953, is a HUA patient.

[0062] Medical history: The patient's uric acid level has been continuously monitored since 2016 and has remained persistently high.

[0063]

[0064] Intervention measures: Since the beginning of 2020, the patient has taken MF 450 mg daily.

[0065] Uric acid monitoring results ( Figure 7 )

[0066]

[0067] Result analysis:

[0068] The uric acid level has decreased significantly: Compared with 2019, the uric acid level in 2024 has decreased by 201 μmol / L.

[0069] Long-term stability: The uric acid level remained stable and gradually decreased during the period of taking MF.

[0070] Safety and tolerance: The patient did not report any adverse reactions or discomfort symptoms.

[0071] Case 2: A case study on the comparison of the effects of taking MF and drugs is as follows:

[0072] Medical history: Male patient B, born in 1966, is a patient with HUA and gout. He has had gout for more than 10 years. The uric acid value was 520 μmol / L during the period without taking medicine. The patient has been taking febuxostat 20 mg / day all year round, and the uric acid has been maintained at 360 μmol / L.

[0073] Intervention measures: To compare the effects of taking MF, febuxostat was stopped for 7 days, and the uric acid value returned to 520 μmol / L. Then MF was taken once a day, on an empty stomach, 450 mg each time, for 15 consecutive days. The changes in uric acid values are shown in Figure 8 .

[0074] Uric acid monitoring results: The uric acid value decreased significantly on the 2nd day of taking the medicine. The uric acid value had dropped to 380 μmol / L on the 13th day of taking the medicine, which was lower than the critical value (420 μmol / L). After 15 days, MF was stopped, and the uric acid value rose to 520 μmol / L. The patient did not report any adverse reactions or discomfort symptoms. The data showed that the effect of MF was comparable to that of febuxostat in reducing uric acid levels.

[0075] Case 3-12: For the gender, age, dosage, administration time, and uric acid levels before and after administration in Case 3-12, please refer to Table 2. Among them, 4 patients took the drug 3 times a day, 450 mg each time, and the uric acid level decreased by 120 - 340 μmol / L before and after the administration period; 5 patients took the drug 2 times a day, 450 mg each time, and the uric acid level decreased by 60 - 139 μmol / L before and after the administration period; 1 patient took the drug 1 time a day, 450 mg each time, and the uric acid level decreased by 30 μmol / L before and after the administration period.

[0076] Table 2 Changes in Uric Acid Levels of 10 Subjects before and after Taking MF

[0077]

[0078] For the above 10 subjects taking MF, the uric acid level decreased significantly, and no adverse reactions or discomfort symptoms were reported, demonstrating its uric acid-lowering effect and safety during long-term use, and it can be used as an effective means for treating HUA and gout.

[0079] Example 3

[0080] Mechanism of action of MF in lowering uric acid and treating gout:

[0081] Using artificial intelligence technology, through molecular docking, molecular dynamics simulation, enhanced sampling techniques, and quantum mechanics / molecular mechanics (QM / MM) simulation, the mechanism of action of MF in lowering uric acid was studied, mainly including:

[0082] 2.1 Inhibiting the function of renal uric acid transporter (URAT1) in the kidney

[0083] High binding affinity: Through simulation, the binding free energy of MF to URAT1 is approximately -40 kcal / mol, higher than that of known URAT1 inhibitors (such as benzbromarone and probenecid). The binding free energy of MF to GLUT9 is approximately -35 kcal / mol, showing a strong binding ability.

[0084] Key interactions: MF binds to the key sites of these transporters (such as Arg477 and Lys35 of URAT1, 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 group of MF forms hydrogen bonds with the protein, jointly improving the binding stability.

[0085] Action effect: Inhibiting URAT1 and GLUT9 can significantly reduce the reabsorption of uric acid in the renal tubules, increase the excretion of uric acid, and thus lower the serum uric acid level.

[0086] 2.2 Auxiliary inhibition of the activity of xanthine oxidase (XO) in the liver

[0087] Binding affinity: The sulfate group of MF forms strong electrostatic interactions and hydrogen bonds with the key residues (such as Glu802 and Arg880) of the active site of XO. 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).

[0088] Limited effect: It has a weak inhibitory effect on XO, and may only partially reduce the production of uric acid, playing an auxiliary role in reducing uric acid levels.

[0089] 2.3 Effects on other uric acid transporters

[0090] Low binding affinity: The binding free energy of MF with proteins such as OAT1, OAT3, and ABCG2 is about -25 kcal / mol, with a weak effect.

[0091] Limited impact: It has no significant effect on the secretion and excretion of uric acid, and has a relatively small effect on reducing uric acid levels.

[0092] 2.4 Reduction of inflammatory response

[0093] The occurrence of HUA and gout is closely related to oxidative stress. The excessive accumulation of uric acid can lead to the generation of free radicals, causing tissue damage. MF has 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 alleviating inflammation and oxidative stress and improving the tissue microenvironment.

Claims

1. Application of Undaria pinnatifida spore leaf fucoidan in the preparation of medicines and foods for reducing uric acid; the basic structure of the Undaria pinnatifida spore leaf fucoidan is as follows: ； The number average molecular weight of the spore leaf fucoidan of Undaria pinnatifida is 95,000, the weight average molecular weight is 437,000, the fucose content is 21.0%, the galactose content is 20.6%, and the sulfate content is 25.3%.

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

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

4. The use according to claim 1, characterized in that: The active ingredient in the 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, 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 dosage forms of the medicine include oral liquid, capsule, tablet and granule.

9. The use according to claim 1, characterized in that: The dosage form of the drug is a targeted agent.

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