Application of aminoguanidine compounds in the preparation of drugs for treating gout caused by gosling astrovirus type 2

Abstract

This invention relates to the application of aminoguanidine compounds in the preparation of drugs for treating gout, primarily in the preparation of drugs for treating gout caused by gosling astrovirus type 2. The dosage of aminoguanidine hydrochloride or aminoguanidine sulfate in the drug is 5–25 mg / day / gosling. The application of aminoguanidine compounds in the preparation of drugs for treating gout provided in the above technical solution can effectively reduce gosling mortality, alleviate joint swelling and urate deposition in viscera, reduce the content of nitric oxide and nitric oxide synthase in kidney tissue, and reduce the content of pro-inflammatory factors IL-1β and IL-8, inhibiting autophagy in renal tubular epithelial cells. In other words, aminoguanidine compounds can effectively alleviate gout caused by gosling astrovirus type 2.

Description

Technical Field

[0001] This invention relates to the field of veterinary drug technology, specifically the application of aminoguanidine compounds in the preparation of drugs that alleviate gout symptoms in goslings caused by type 2 goose astrovirus and reduce gosling mortality. Background Technology

[0002] A new disease has emerged in goose farms, characterized primarily by urate deposits in the internal organs and joints. It is mainly caused by goose astrovirus type 2 (GAstV-2), resulting in mass mortality of goslings. Currently, there is no commercially available vaccine to prevent goose astrovirus infection. Research institutions are developing hyperimmune egg yolk antibodies to treat gosling gout, but the treatment efficacy is poor due to strain differences and low antibody titers. Furthermore, subcutaneous or intramuscular injections cause significant stress to the goslings, leading to mass mortality within 1-2 days after antibody injection.

[0003] Currently, there is still a lack of safe and effective veterinary drugs for the prevention and treatment of gout in goslings. Summary of the Invention

[0004] This invention addresses the current lack of commercially available veterinary drugs for treating gout in goslings. Based on research into the mechanism of kidney damage caused by goose astrovirus type 2, the inventors developed aminoguanidine, a nitric oxide inhibitor that alleviates gout symptoms and reduces gosling mortality. Aminoguanidine primarily alleviates gout symptoms and reduces gosling mortality by inhibiting the production of nitric oxide and pro-inflammatory factors in the gosling kidneys and reducing autophagy in renal tubular epithelial cells.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] This invention relates to the application of aminoguanidine compounds in the preparation of drugs for treating gout caused by gosling astrovirus type 2.

[0007] The aminoguanidine compound in the drug is aminoguanidine hydrochloride or aminoguanidine sulfate.

[0008] Preferably, the drug may or may not contain other active ingredients.

[0009] Preferably, the drug does not contain other active ingredients, but contains pharmaceutically acceptable excipients.

[0010] Preferably, the dosage of aminoguanidine hydrochloride or aminoguanidine sulfate in the drug is 5–25 mg / day / animal.

[0011] The above-mentioned technical solution describes the application of aminoguanidine compounds in the preparation of drugs for treating gout. When using this method, each 2-day-old healthy gosling is orally infected with 0.8 mL of type 2 goose astrovirus GAstV-2 (TCID2). 50 =10 -4.69 The animal infection model was replicated by injecting 0.1 mL of aminoguanidine hydrochloride (i.e., 5 mg and 25 mg / bird / day) into the peritoneum on the same day. The aminoguanidine hydrochloride was then injected continuously for 7 days. During this period, the clinical symptoms and mortality of the goslings were observed. After the medication was completed, the goslings were euthanized to observe kidney lesions. The levels of nitric oxide and nitric oxide synthase, changes in pro-inflammatory factors, and changes in autophagy-related molecules in the kidney tissue of the goslings were detected.

[0012] The results showed that treatment with aminoguanidine compounds could effectively reduce gosling mortality, alleviate joint swelling and urate deposition in viscera, reduce the levels of nitric oxide and nitric oxide synthase in kidney tissue and the levels of pro-inflammatory factors IL-1β and IL-8, and inhibit the occurrence of autophagy in renal tubular epithelial cells. In other words, aminoguanidine compounds can effectively alleviate gout caused by gosling astrovirus type 2. Attached Figure Description

[0013] Figure 1 A graph showing the reduction of spleen and kidney organ coefficients in GAstV-2 infected goslings by aminoguanidine;

[0014] Figure 2 Organ lesions in goslings infected with GAstV-2 were illustrated using aminoguanidine to alleviate the symptoms.

[0015] Figure 3 Pathological image of kidney tissue damage in goslings infected with GAstV-2, as a result of aminoguanidine.

[0016] Figure 4 Pathological image of liver tissue damage in goslings infected with GAstV-2, as a result of aminoguanidine.

[0017] Figure 5 Aminoguanidine can reduce serum uric acid and creatinine levels in GAstV-2 infected goslings;

[0018] Figure 6 A graph showing how aminoguanidine reduces the levels of pro-inflammatory factors in the kidneys of GAstV-2 infected goslings.

[0019] Figure 7 A graph showing how aminoguanidine reduces autophagy levels in the kidneys of GAstV-2 infected goslings. Detailed Implementation

[0020] To make the objectives and advantages of this invention clearer, the invention will be specifically described below with reference to embodiments. It should be understood that the following text is merely used to describe one or more specific embodiments of the invention and does not strictly limit the scope of protection specifically claimed by the invention.

[0021] Example

[0022] (1) Preparation of aminoguanidine hydrochloride solution

[0023] Aminoguanidine hydrochloride (AG) was dissolved in PBS to prepare solutions with concentrations of 10 mg / mL and 50 mg / mL, stored at 4°C, and equilibrated to room temperature before use.

[0024] (2) Grouping of experimental animals

[0025] One-day-old goslings were randomly divided into four groups of eight each. Except for the control group, all goslings in each group were orally infected with GAstV-2 virus solution at two days of age.

[0026] ①Control group: Starting from day 2, 0.5 mL of PBS was injected intraperitoneally daily for 7 consecutive days.

[0027] ②GAstV-2 group: Starting from day 2, 0.5 mL of PBS was injected intraperitoneally daily for 7 consecutive days.

[0028] ③AG (5mg / d) group: Starting from day 2, 0.5mL of AG solution with a concentration of 10mg / mL was injected intraperitoneally every day for 7 consecutive days.

[0029] ④AG (25mg / d) group: Starting from day 2, 0.5mL of AG solution with a concentration of 50mg / mL was injected intraperitoneally every day for 7 consecutive days.

[0030] (3) Clinical symptom statistics and autopsy sampling

[0031] Daily observations and statistics were conducted on the mental state, number of deaths, and number of goslings with standing difficulties in each group. On the eighth day after viral infection, necropsies were performed on goslings in each group to observe and analyze urate deposition in the ureters, joint cavities, and internal organs. The weight of each gosling, as well as the weight of its spleen and kidneys, were measured. A portion of the liver and kidneys was collected and fixed in paraformaldehyde for subsequent pathological section preparation; the other portion was stored in cryovials at -80°C for subsequent detection of inflammatory factors and autophagy levels. Blood was collected from the wing vein, centrifuged at 1500 rpm for 15 minutes to separate serum, and creatinine and uric acid were measured to assess kidney damage.

[0032] The test results are as follows:

[0033] (1) Aminoguanidine reduced mortality and clinical symptoms in goslings infected with GAstV-2, as shown in Table 1.

[0034] Table 1. Aminoguanidine reduces mortality and clinical symptoms in goslings infected with GAstV-2.

[0035]

[0036] (2) Aminoguanidine reduces the organ coefficients of the kidneys and spleen in GAstV-2 infected goslings.

[0037] like Figure 1 As shown, the organ coefficients of the kidneys and spleen increased in goslings infected with GAstV-2, indicating that GAstV-2 infection leads to swelling of the spleen and kidneys in goslings. After using AG, the organ coefficients of the spleen and kidneys in goslings decreased significantly, indicating that AG can alleviate the swelling of the spleen and kidneys in goslings after GAstV-2 infection.

[0038] (3) Aminoguanidine alleviates organ lesions in goslings infected with GASTV-2.

[0039] Figure 2 Anatomical photographs of the liver, kidneys, gallbladder, and joint cavity are shown. As can be seen from the figures, there are a large amount of urate deposits in the liver, kidneys, gallbladder, and joint cavity of goslings in the GAstV-2 group. The urate deposits in the viscera and joint cavity of goslings in the low-dose AG group (5mg / d) are less than those in the GAstV-2 group. In the high-dose AG group (25mg / d), almost no obvious urate deposits are observed on the surface of the viscera and in the joint cavity of goslings.

[0040] (4) Aminoguanidine reduces the pathological damage to the kidney tissue of goslings infected with GAstV-2.

[0041] Depend on Figure 3 It can be seen that the kidney tissue structure in the control group is clear, the renal tubular structure is intact, and the brush border is present in the proximal tubules;

[0042] In the GAstV-2 group, diffuse infiltration of inflammatory cells was observed (indicated by thick gray arrows), destruction of the brush border of the proximal convoluted tubule (indicated by thick black arrows), necrosis of some renal tubular epithelial cells (indicated by thick white arrows), and a large number of sloughed renal tubular epithelial cells and protein casts were found in the lumen of the renal tubules (indicated by thin black arrows).

[0043] The renal tubular structure of the AG group (25mg / d) was intact, the brush border was clear, and the infiltration of inflammatory cells was significantly reduced compared with the GAstV-2 group.

[0044] (5) Aminoguanidine reduces liver tissue pathological damage in GAstV-2 infected goslings.

[0045] from Figure 4 It can be seen that the liver tissue morphology of the control group was normal, the hepatic cords were neatly arranged, and no obvious lesions were observed;

[0046] In the GAstV-2 group, diffuse infiltration of numerous inflammatory cells was observed (indicated by thick black arrows), some hepatocytes showed fatty degeneration (indicated by thin black arrows), and scattered hemorrhages were observed in the interstitium (indicated by thick white arrows).

[0047] The AG group (25mg / d) showed scattered, small amounts of inflammatory cell infiltration, and the degree of lesion was significantly milder than that of the GAstV-2 group.

[0048] (6) Aminoguanidine reduces serum uric acid and creatinine levels in GAstV-2 infected goslings.

[0049] Excessive uric acid is a major cause of gout in goslings, and serum uric acid and creatinine levels are also important indicators of kidney function. Figure 5 It can be seen that, compared with the control group, the concentrations of uric acid and creatinine in the serum of goslings in the GAstV-2 group were significantly increased, indicating that GAstV-2 causes kidney damage. After injection of AG, the levels of uric acid and creatinine were significantly reduced compared with the GAstV-2 group, indicating that AG can effectively alleviate the kidney damage caused by GAstV-2.

[0050] (7) Aminoguanidine reduces the level of pro-inflammatory factors in the kidneys of GAstV-2 infected goslings.

[0051] from Figure 6 It can be seen that infection with GAstV-2 significantly increased the expression levels of pro-inflammatory factors iNOS, IL-1β, IL-8, and TNF-α in the kidneys of goslings, as well as the concentration of nitric oxide in the serum. The high expression of pro-inflammatory factors and excessive nitric oxide may trigger inflammation, leading to kidney damage. Intraperitoneal injection of AG significantly decreased the expression levels of pro-inflammatory factors iNOS, IL-1β, and IL-8, and also significantly reduced the concentration of NO in the serum.

[0052] (8) Aminoguanidine reduces autophagy levels in the kidneys of GAstV-2 infected goslings.

[0053] ATG5, Beclin1, and LC3B are key indicators of autophagy. From Figure 7 It can be seen that the expression levels of ATG5, Beclin1, and LC3B in the kidneys of goslings were all upregulated after GAstV-2 infection, indicating that GAstV-2 induced autophagy in the kidneys of goslings. After intraperitoneal injection of AG, the expression levels of ATG5 and Beclin1 were significantly downregulated, and the expression of LC3B also showed a decreasing trend, indicating that AG can reduce the level of autophagy in the kidneys of GAstV-2 infected goslings.

[0054] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments. For those skilled in the art, after learning the contents described in the present invention, several equivalent changes and substitutions can be made without departing from the principle of the present invention. These equivalent changes and substitutions should also be considered to fall within the protection scope of the present invention.

Claims

1. Use of aminoguanidine hydrochloride in the preparation of a medicament for treating gosling gout caused by goose astrovirus type 2.

2. Use according to claim 1, characterized in that: The medicament can or can not include other active ingredients.

3. Use according to claim 1, characterized in that: The medicament contains pharmaceutically acceptable adjuvants and does not contain other active ingredients.

4. Use according to claim 1, characterized in that: The aminoguanidine hydrochloride is used in a dosage of 5-25 mg / day / bird.

Images