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Building method for acute hyperuricemia animal model

A technology of hyperuricemia and construction method, which is applied in the field of construction of animal models of acute hyperuricemia, can solve the problems of unsuitable PNP enzyme inhibitors for reducing uric acid, and achieve good reproducibility and simple construction method

Active Publication Date: 2017-05-10
KPC PHARM INC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing commercially available drugs that inhibit uric acid production all target xanthine oxidase (XO). From the perspective of uric acid production pathway, inosine forms uric acid through the sequential hydrolysis and oxidation of PNP enzyme and XO enzyme. That is, the PNP enzyme is located upstream of the XO enzyme, so the animal model of hyperuricemia established by supplementing purine drugs (food) is not suitable for evaluating the uric acid-lowering effect of PNP enzyme inhibitors

Method used

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  • Building method for acute hyperuricemia animal model
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  • Building method for acute hyperuricemia animal model

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Experimental program
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Effect test

Embodiment 1

[0019] 1. Experimental reagents

[0020] Uric acid (UA) assay kit: 50T / 48 samples, batch number: 20160701, Nanjing Jiancheng Bioengineering Research Institute; sodium chloride injection: 250ml, 2.25g, batch number: A140829G2, Kunming Nanjiang Pharmaceutical Co., Ltd. Inosine: 14125-10G, Lot #019K1124V, CAS: 58-63-9, Sigma.

[0021] 2. Experimental animals

[0022] Male rhesus monkey, weighing 6-8kg, production license number: SCXK (Dian) 2011-0005, use license number: SYXK (Dian) K2014-007.

[0023] 3. Experimental equipment

[0024] SYNERGY / HT multifunctional microplate reader, BioTek; MiniSpin centrifuge, eppendorf; NewClassic MFMS105 analytical balance, METTLER TOLEDO.

[0025] 4. Experimental method

[0026] Four rhesus monkeys were divided into four groups: model group and normal saline control group. The model group was intraperitoneally given different doses of inosine solution to create models. High dose group (100mg / kg), middle dose group (50mg / kg), low dose gro...

Embodiment 2

[0033] After a time interval of two weeks, the 4 rhesus monkeys used in the experimental group of Example 1 were used, 2 new male rhesus monkeys were added, and 6 monkeys were used to build a model, and the blood uric acid level was used as a control. The dose of inosine is (100mg / kg), intraperitoneally injected, and the administration volume is 2ml / kg. About 2ml of blood was taken at different time points, and the blood uric acid value was compared at different time points. The time points were set as: 0h (that is, blank control blood was taken before modeling), 0.5h, 1h, 2h, 4h, and 8h after modeling. After the blood samples were left at room temperature for 30 minutes, they were centrifuged at 4000r / min for 10 minutes, and the serum was taken and immediately frozen in a -80°C refrigerator. After the serum was collected at all time points, the uric acid value was determined according to the kit method (phosphotungstic acid method). The results are shown in Table 2, figure ...

Embodiment 3

[0039] Six male rhesus monkeys were divided into two drug groups. 3 only for Ulodesine group, 3 only for Febuxostat group. Drugs were given 1 hour before modeling. The dose of Ulodesine was 2 mg / kg (equivalent to clinical human dose), and the dose of Febuxostat was 2 mg / kg (equivalent to clinical human dose). The dosage of the modeling agent inosine is (100mg / kg), injected intraperitoneally, and the administration volume is 2ml / kg. Take about 2ml of blood at different time points, and compare the blood uric acid values ​​at different time points. 4h, 8h, and 24h after administration. After the blood samples were left at room temperature for 30 minutes, they were centrifuged at 4000r / min for 10 minutes, and the serum was taken and immediately frozen in a -80°C refrigerator. After the serum was collected at all time points, the uric acid value was determined according to the kit method (phosphotungstic acid method). The results are shown in Table 3, image 3 and Figure 4 ,...

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Abstract

The invention provides a building method for an acute hyperuricemia animal model. The building method comprises the following steps: applying inosine to a rhesus monkey to obtain an acute hyperuricemia animal model. In the building method, the inosine is taken as an inducer for inducing rise of serum uric acid, and is applied to the rhesus monkey to build the hyperuricemia animal model for assessing the efficacy of decreasing serum uric acid. As proved by experiment results, the acute hyperuricemia animal model can be built by the method provided by the invention, and the model has high reproducibility; the hyperuricemia animal model built by the method can be used for evaluating the serum uric acid decreasing function of a PNP enzyme inhibitor, and is suitable for assessing the serum uric acid decreasing effect of an XO inhibitor. Moreover, the building method provided by the invention is simple, only injection of inosine into the rhesus monkey is required, and other medicaments are not required.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a method for constructing an animal model of acute hyperuricemia. Background technique [0002] Hyperuricemia is a pathological state in which urate in the blood exceeds the normal level (6 mg / 100 ml) due to excessive production of uric acid caused by various factors, or decreased excretion of uric acid by the kidneys, or both. Hyperuricemia is the most important biochemical basis of gout, and it is closely related to cardiovascular diseases and immune system disorders, which seriously endangers human health. The drugs currently used clinically to treat hyperuricemia mainly inhibit uric acid production and promote uric acid excretion. With the rapid development of molecular biotechnology and people's deepening understanding of the pathogenesis and genetics of hyperuricemia, new therapeutic targets have been discovered, and new therapeutic drugs have also come out one after anot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7076
Inventor 张建文黄茜闫东明杨旭娟李鹏辉
Owner KPC PHARM INC
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