Use of salvianic acid a or pharmaceutically acceptable salt thereof in preparing drug or health-care product for preventing and treating hyperuricemia, gout, and uric acid nephropathy

Abstract

Disclosed in the present invention is use of salvianic acid A or a pharmaceutically acceptable salt thereof in preparing a drug or health-care product for preventing and treating hyperuricemia, gout, and uric acid nephropathy. Animal experiments prove that salvianic acid A can greatly reduce the blood uric acid level, and meanwhile, ameliorate the degree of renal tubule dilation, necrosis, and renal fibrosis of hyperuricemia rats.

Description

The use of tanshinone or its drug salts in the preparation of drugs or health products for the prevention and treatment of hyperuricemia, gout, and uric acid nephropathy. Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology related to tanshinone, specifically relating to the application of tanshinone or its pharmaceutically acceptable salt in the preparation of drugs or health products for the prevention and treatment of hyperuricemia, gout and uric acid nephropathy. Background Technology

[0002] Hyperuricemia (HUA) is defined as a fasting serum uric acid level >7.0 mg / dL (420 μmol / L) in men and >6.0 mg / dL (360 μmol / L) in women on two separate days under normal purine dietary conditions. Hyperuricemia is a chronic metabolic disease caused by impaired purine and uric acid metabolism, and is an important biochemical basis for diseases such as gout and uric acid nephropathy. Currently, the prevalence of HUA in Chinese adults is 14.1%, and the prevalence is increasing year by year and showing a trend towards affecting younger people. Due to my country's large population, the number of people with HUA reaches 190 million, making it the "fourth highest disease" after hypertension, hyperglycemia, and hyperlipidemia, and the second most common metabolic disease after diabetes. It seriously threatens human life and health, and brings enormous economic and psychological burdens to society and families. HUA is also a starting factor for many diseases, participating in the occurrence and development of metabolic diseases (diabetes, metabolic syndrome, hyperlipidemia, etc.), chronic kidney disease, and cardiovascular and cerebrovascular diseases.

[0003] Currently, common medications for treating gout include colchicine, nonsteroidal anti-inflammatory drugs (NSAIDs), allopurinol, febuxostat, and benzbromarone. These drugs aim to alleviate and treat gout and hyperuricemia by inhibiting uric acid formation and / or promoting uric acid excretion. However, their clinical application is limited due to adverse reactions. For example, colchicine is highly toxic, and the therapeutic dose is very close to the toxic dose; allopurinol has a serious adverse reaction, exfoliative dermatitis, with a mortality rate as high as 20%–25%; febuxostat increases adverse cardiovascular events and mortality and has been given a black box warning by the U.S. Food and Drug Administration (FDA); and benzbromarone has severe liver and kidney toxicity and has been banned by the FDA. Current clinical guidelines recommend that uric acid-lowering drugs need to be taken long-term or even lifelong. Therefore, finding safe and effective drugs with fewer adverse reactions to treat hyperuricemia has become a pressing issue in this field.

[0004] Traditional Chinese medicine Danshen (Salvia miltiorrhiza Bunge.) has the effects of removing blood stasis and relieving pain, promoting blood circulation and regulating menstruation, and clearing the heart and relieving irritability. Modern pharmacological studies have shown that Danshen has the effects of anti-atherosclerosis, improving blood circulation, inhibiting platelet adhesion and aggregation, scavenging oxygen free radicals in the body, improving hypoxia tolerance, improving coronary artery blood supply, and protecting against damage to heart and brain cells. Studies have shown that Danshen extract significantly reduces uric acid levels in hyperuricemic mice and promotes uric acid excretion in urine. It is a major component of traditional Chinese medicine formulas for treating uric acid nephropathy, such as Jianpi Yishen Fang, Yishen Xiezhuo Tang, Yishen Huoxue Fang, and Jiawei Siwu Tang. Danshensu (Salvianic acid A) is the most important water-soluble component of Danshen. It can reduce the damage to cells caused by inflammatory factors and free radicals through antioxidant, anti-inflammatory, and anti-apoptotic effects, maintain endothelial cell homeostasis, and protect cells. However, there are currently no reports on the effects of tanshensu in lowering uric acid, preventing and treating gout, and treating uric acid nephropathy. Summary of the Invention

[0005] This invention relates to tanshinone or its pharmaceutically acceptable salts, and also to the use of pharmaceutical compositions comprising tanshinone and its pharmaceutically acceptable salts in the preparation of medicaments for the prevention and / or treatment of hyperuricemia, gout, and uric acid nephropathy. Furthermore, it relates to the use of tanshinone and its pharmaceutically acceptable salts in the preparation of medicaments or health products that lower blood uric acid levels. The application of this invention can provide new medication options for the clinical treatment of hyperuricemia and the resulting hyperuricemic nephropathy.

[0006] This invention provides the use of tanshinone or its pharmaceutically acceptable salts in the preparation of drugs or health products for the prevention and treatment of hyperuricemia, gout, and uric acid nephropathy.

[0007] Furthermore, the tanshinone has a structure as shown in formula (1):

[0008] Furthermore, the tanshinone or its pharmaceutically acceptable salt can reduce serum creatinine levels by improving glomerular function.

[0009] Furthermore, the tanshinone or its drug can be used to promote uric acid excretion and lower blood uric acid levels.

[0010] Furthermore, the tanshinone or its drug-acceptable salt can improve glomerular basement membrane, enhance glomerular filtration, and reduce the urine protein-to-creatinine ratio.

[0011] Furthermore, the tanshinone or its pharmaceutically acceptable salts improve renal tubular dilation and / or necrosis by lowering blood uric acid levels, reducing urate deposition, inhibiting inflammation and fibrosis.

[0012] Furthermore, the tanshinone or its drug-acceptable salt can reduce kidney inflammation by inhibiting the TLR4 / NF-κB signaling pathway, inhibiting the activation of the NLRP3 inflammasome, and thus reducing kidney inflammation.

[0013] Furthermore, the tanshinone or its drug-acceptable salts improve renal fibrosis by inhibiting renal tubular epithelial-mesenchymal transition, inhibiting the renin-angiotensin system, inhibiting vascular smooth muscle cell proliferation, inhibiting oxidative stress, and inhibiting autophagy.

[0014] Furthermore, the effective dose of the tanshinone or its pharmaceutically acceptable salt is 1–1000 mg / kg.

[0015] Furthermore, the aforementioned pharmaceutical or health product is a clinically acceptable formulation made by mixing tanshinone or its pharmaceutically acceptable salt as the active ingredient with a pharmaceutically acceptable excipient or carrier.

[0016] Furthermore, the preparation is an oral preparation or an injectable preparation.

[0017] Furthermore, the oral preparation is an oral tablet.

[0018] In this invention, tanshinone or its pharmaceutically acceptable salts refer to acidic and / or basic salts formed by tanshinone with inorganic bases and / or acids, organic bases and / or acids, including zwitterionic salts (internal salts), and quaternary ammonium salts, such as alkyl ammonium salts. The salts described in this invention are selected from: sodium tanshinone, potassium tanshinone, calcium tanshinone, lithium tanshinone, magnesium tanshinone, ammonium tanshinone, meglumine tanshinone, amine tanshinone, arginine tanshinone, and lysine tanshinone.

[0019] The advantages of this invention compared to the prior art are as follows:

[0020] This invention provides the use of tanshinone or its salts in the preparation of drugs for lowering serum uric acid levels and preventing and treating uric acid nephropathy. Animal experiments have demonstrated that tanshinone can significantly reduce serum uric acid levels while improving the degree of renal tubular dilation, necrosis, and renal fibrosis in hyperuricemic mice. The application of this invention can provide new drug options for the clinical treatment of hyperuricemia and its resulting hyperuricemic nephropathy. Attached Figure Description

[0021] Figure 1 shows the results of serum uric acid level detection in rats in Example 1 (Note: *** vs normal group P<0.001, # vs model group P<0.05, ### vs model group P<0.001);

[0022] Figure 2 shows the results of serum creatinine level detection in rats in Example 1 (Note: *** vs normal group P<0.001, ## vs model group P<0.01,### vs model group P<0.001);

[0023] Figure 3 shows the results of the urine protein-to-creatinine ratio detection in rats in Example 1 (Note: *** vs normal group P<0.001, ## vs model group P<0.01, ### vs model group P<0.001);

[0024] Figure 4 shows the PAS staining of rat kidney tissue in Example 1;

[0025] Figure 5 shows Masson staining of rat kidney tissue in Example 2;

[0026] Figure 6 shows the quantitative collagen volume in rat kidneys obtained by Masson staining in Example 2 (Note: *** vs. normal group P<0.001, ### vs model group P<0.001);

[0027] Figure 7 shows the immunohistochemical staining of rat kidney α-SMA in Example 2;

[0028] Figure 8 shows the percentage of the area of ​​the α-SMA immunohistochemically positive region in the rat kidney in Example 2.

[0029] Figure 9 shows the expression of renal α-SMA protein in Example 2 using Western blot analysis;

[0030] Figure 10 shows the quantitative analysis of renal α-SMA protein in Example 2 (Note: *** vs normal group P<0.001, ### vs model group P<0.001);

[0031] Figure 11 shows the 24-hour urinary uric acid excretion in mice in Example 2 (Note: *** vs normal group P<0.001, # vs model group P<0.05, ### vs model group P<0.001). Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] As used herein, the term "tanshinone or its pharmaceutically acceptable salt" refers to acidic and / or basic salts formed by tanshinone with inorganic bases and / or acids, organic bases and / or acids, including zwitterionic salts (internal salts), and quaternary ammonium salts, such as alkyl ammonium salts. The salts described in this invention are selected from: sodium tanshinone, potassium tanshinone, calcium tanshinone, lithium tanshinone, magnesium tanshinone, ammonium tanshinone, meglumine tanshinone, amine tanshinone, arginine tanshinone, and lysine tanshinone.

[0034] The tanshinone of this invention can be administered to patients in the form of a pharmaceutically acceptable salt or pharmaceutical composition. A complex needs to be mixed with a suitable carrier or excipient to form a pharmaceutical composition to ensure an effective therapeutic dose. "Effective therapeutic dose" refers to the dose necessary for tanshinone or its pharmaceutically acceptable salt to achieve a therapeutic effect.

[0035] Tanshinone or its pharmaceutically acceptable salts can be formulated into various dosage forms, including solid, semi-solid, liquid, and aerosol formulations (Remington's Pharmaceutical Sciences, Mack Publishing Company (1995), Philadelphia, PA, 19th ed.). Specific dosage forms within these categories include tablets, pills, sugar tablets, granules, gels, ointments, solutions, suppositories, injections, inhalers, and sprays. These dosage forms can be used for both local and systemic administration, as well as for immediate-release or sustained-release administration.

[0036] When tanshinone or its pharmaceutically acceptable saline is administered by injection, these compounds can be formulated into solutions, suspensions, and emulsions using water-soluble or lipid-soluble solvents. Lipid-soluble solvents specifically include vegetable oils and similar oils, synthetic fatty acid glycerides, higher fatty acid esters, and proylene glycol esters. These compounds are more readily soluble in ethanol solutions and trace amounts of DMSO solutions.

[0037] When tanshinone or its pharmaceutically acceptable saline is administered orally, it can be compounded with pharmaceutically acceptable excipients using common techniques. These excipients can formulate these compounds into various dosage forms that can be used by patients, such as tablets, pills, suspensions, and gels. There are several methods for formulating oral dosage forms, such as first mixing the compound and solid excipients, thoroughly grinding the mixture, adding appropriate excipients, and processing it into granules. Excipients that can be used to formulate oral dosage forms include: sugars such as lactose, sucrose, mannitol, or sorbitol; and celluloses such as corn starch, wheat starch, potato starch, gelatin, taro gum, methylcellulose, hydroxymethylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone.

[0038] The tanshinone or its pharmaceutically acceptable salts involved in this invention can also be formulated as a spray, which is achieved via a pressurizer and a sprayer or a dry powder inhaler. Suitable propellants that can be used in the sprayer include dichlorodifluoromethane, chloroform, dichlorotetrafluoroethane, carbon dioxide, and dimethyl ether. The dosage of the aerosol can be adjusted via a valve on the sprayer.

[0039] The various dosage forms involved in this invention relate to the effective therapeutic dose of tanshinone or its pharmaceutically acceptable salts. The effective therapeutic dose of these compounds depends on the patient receiving treatment. In determining the appropriate dose, the patient's weight, condition, method of administration, and the prescribing physician's subjective judgment must be taken into account. The therapeutically effective amount of tanshinone or its pharmaceutically acceptable salts, and compositions containing these compounds, should be determined by a competent and experienced prescribing physician.

[0040] Although the effective therapeutic dose of tanshinone or its pharmaceutically acceptable salts can vary depending on the patient’s condition, the usual appropriate dosage range is 1–1000 mg / kg.

[0041] Example 1: Tanshinone reduces serum uric acid levels and improves kidney damage

[0042] Animal modeling and grouping for drug administration, the specific steps are as follows:

[0043] Experimental animals: Sixty SPF-grade male SD rats were randomly divided into 6 groups of 10 each, weighing 180–220 g. They were fed a standardized experimental diet under constant temperature conditions for at least one week prior to the experiment. The rats were housed in separate cages with free access to water and food at a room temperature of (25±2)℃ and a relative humidity of 50%–70%.

[0044] Animal grouping: Sixty rats were divided into six groups: normal control group, hyperuricemia model group, low-dose tanshinone group (5mg / kg / d), medium-dose tanshinone group (10mg / kg / d), high-dose tanshinone group (50mg / kg / d), and positive control allopurinol group (50mg / kg / d), with 10 rats in each group.

[0045] Drugs and reagents: potassium oxonate (Sigma); adenine (Sigma); allopurinol (Shanghai Titan Technology Co., Ltd.); tanshinone (Shanghai Zhongshi Pharmaceutical Co., Ltd.).

[0046] Experimental Methods: Establishment, drug administration, and material collection methods for the animal model of hyperuricemia. Except for the normal control group, all other groups were administered adenine 1.5 g / kg and potassium oxalate 1.5 g / kg daily by gavage to establish a hyperoxalemia model for 21 consecutive days. The tanshinone and allopurinol treatment groups received tanshinone and allopurinol by gavage daily for 21 consecutive days. The normal control group and the model group received the same volume of physiological saline.

[0047] Biochemical index determination: After the last administration, rats were placed in metabolic cages, fasted but allowed free access to water, and urine was collected for 24 hours. After urine collection, blood was collected from the eyeballs of rats in each group. Blood samples were centrifuged at 3000 r / min for 15 min at room temperature, and serum was collected for biochemical index determination using a fully automated biochemical analyzer. Mouse urine samples were centrifuged at 3500 r / min for 10 min at room temperature, and the supernatant urine was collected for biochemical index determination using a fully automated biochemical analyzer. After blood collection, both kidneys were removed, paraffin sections were prepared using paraformaldehyde, and PAS staining was performed to observe kidney structure. The experimental results are shown in Figure 4. MASSON staining was used to observe renal fibrosis. The experimental results are shown in Figure 5.

[0048] As shown in Figure 1, compared with the normal group, the model group had significantly higher uric acid levels, indicating that the combination of adenine and potassium oxonate can induce hyperuricemia in rats. Tanshinone can significantly reduce the levels of serum uric acid, creatinine, and the ratio of urinary protein to creatinine, and the high-dose treatment group can achieve better results than allopurinol.

[0049] As shown in Figure 2, high and medium doses of tanshinone significantly reduced serum creatinine levels, and the effect was better than that of allopurinol, indicating that tanshinone has a protective effect on the kidney function of animals while lowering uric acid.

[0050] As shown in Figure 3, tanshinone is more effective than allopurinol in reducing the ratio of urinary protein to creatinine, and it has the effect of improving renal function.

[0051] As shown in Figure 4, compared with the normal group, the model group mice exhibited abnormal kidney structure, including missing brush borders in the renal tubules, glomerular mesangial proliferation, and increased inflammatory cell infiltration in the tubulointerstitium. After administration of tanshinone, renal glycogen deposition was significantly reduced, and glomerular and tubulointerstitial damage was significantly alleviated, with effects exceeding those of the allopurinol group. This indicates that tanshinone can improve renal pathological damage caused by hyperuricemia.

[0052] Example 2: The therapeutic effect of tanshinone on renal fibrosis in rats with hyperuricemia

[0053] Animals were grouped and administered the same as in Example 1. MASSON staining and α-SMA (α-smooth muscle actin) immunohistochemical staining were used to observe the expression of renal collagen fibers and assess the renal fibrosis status. The experimental results are shown in Figures 5-10.

[0054] As shown in Figures 5-6, compared with the normal group, the model group mice showed obvious blue staining in the glomeruli and renal tubules, indicating that there was more collagen deposition and obvious fibrosis. Compared with the model group, the allopurinol and tanshinone administration groups showed varying degrees of reduction, with the high-dose tanshinone group showing a more significant effect.

[0055] α-SMA is a biomarker of fibrosis. As shown in Figure 7 (immunohistochemistry), α-SMA expression was increased in the model group (brown staining). Figure 9 (Weighted Blot) also shows increased α-SMA expression in the model group, indicating that hyperuricemia leads to renal fibrosis. After treatment with tanshinone, α-SMA expression was inhibited, and the degree of renal interstitial fibrosis in hyperuricemic mice was significantly improved. Immunohistochemical staining of α-SMA indicates that tanshinone can effectively inhibit interstitial α-SMA expression, demonstrating that tanshinone can improve renal fibrosis in hyperuricemic rats.

[0056] Example 3: Tanshinone reduces blood uric acid levels by promoting renal uric acid excretion.

[0057] On day 21 of the experiment, 24-hour urine samples from mice were collected using metabolic cages, and the urine volume was recorded. After centrifugation at 800 g / min for 10 min at room temperature, the supernatant urine was collected and its uric acid level was measured using an automated biochemical analyzer. The 24-hour urinary uric acid excretion was calculated as: 24-hour urine volume × 24-hour urinary uric acid concentration. The experimental results are shown in Figure 11.

[0058] As shown in Figure 11, the 24-hour urinary uric acid excretion in the model group mice was significantly reduced; after treatment with tanshinone, urinary uric acid excretion was significantly increased, and the high-dose group was superior to the low-dose group. These results indicate that tanshinone reduces serum uric acid levels by promoting renal uric acid excretion and alleviates the damage to renal tissue caused by urate deposition, thereby protecting renal function.

[0059] This document describes the compounds, their preparation methods, and applications of the present invention in conjunction with specific embodiments and examples, and also sets forth and explains many details. However, it should be understood that the specific embodiments and examples provided herein are merely exemplary and do not limit the scope of protection of the present invention. In fact, those skilled in the art will recognize that the present invention can be implemented in other specific ways, and any modifications, alterations, or adjustments made accordingly do not depart from the spirit and intent of the present invention, and therefore should all be considered to be included within the scope of the present invention.

Claims

1. The use of tanshinone or its drug-acceptable salt in the preparation of drugs or health products for the prevention and treatment of hyperuricemia, gout and uric acid nephropathy.

2. The application according to claim 1, characterized in that: The tanshinone has the structure shown in formula (1):

3. The application according to claim 1, characterized in that: The tanshinone or its drug-acceptable salts reduce serum creatinine levels by improving glomerular function.

4. The application according to claim 1, characterized in that: The tanshinone or its drug can be used to promote uric acid excretion and lower blood uric acid levels.

5. The application according to claim 1, characterized in that: The tanshinone or its drug-acceptable salts improve glomerular basement membrane, enhance glomerular filtration, and reduce the urine protein-to-creatinine ratio.

6. The application according to claim 1, characterized in that: The tanshinone or its drug-acceptable salts improve renal tubular dilation and / or necrosis by lowering blood uric acid levels, reducing urate deposition, inhibiting inflammation and fibrosis.

7. The application according to claim 1, characterized in that: The tanshinone or its drug-acceptable salts reduce kidney inflammation by inhibiting the TLR4 / NF-κB signaling pathway, suppressing the activation of the NLRP3 inflammasome, and thus reducing kidney inflammation.

8. The application according to claim 1, characterized in that: The tanshinone or its drug-acceptable salts improve renal fibrosis by inhibiting renal tubular epithelial-mesenchymal transition, inhibiting the renin-angiotensin system, inhibiting vascular smooth muscle cell proliferation, inhibiting oxidative stress, and inhibiting autophagy.

9. The application according to claim 1, characterized in that: The effective dose of the tanshinone or its acceptable drug salt is 1–1000 mg / kg.

10. The application according to claim 1, characterized in that: The aforementioned medicine or health product is a clinically acceptable formulation made by mixing tanshinone or its pharmaceutically acceptable salt as the active ingredient with a pharmaceutically acceptable excipient or carrier.

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