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Anti-gout active polypeptide RDP2 as well as preparation method and application thereof

An active polypeptide and anti-gout technology, applied in the field of biomedicine, to achieve the effect of strong anti-gout activity, simple structure, and strong uric acid-lowering activity

Active Publication Date: 2020-08-14
KUNMING MEDICAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing clinical anti-gout drugs basically start from one side, either to control the uric acid level during its occult period, or to inhibit the occurrence of inflammation during its attack period. The development of overall anti-gout drugs with dual effects still remains lots of white space

Method used

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  • Anti-gout active polypeptide RDP2 as well as preparation method and application thereof
  • Anti-gout active polypeptide RDP2 as well as preparation method and application thereof
  • Anti-gout active polypeptide RDP2 as well as preparation method and application thereof

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preparation example Construction

[0027] The preparation method of the anti-gout active polypeptide RDP2 of the present invention comprises the following steps:

[0028] A. Soak the rice overnight in deionized water, vacuum freeze-dry the filtered supernatant to obtain material a and store it at -80°C for later use;

[0029] B. Dissolving material a in water to obtain material b, eluting material b with Tris-HCl buffer, collecting the eluent to obtain material c;

[0030] C, subjecting material c to high performance liquid chromatography reversed-phase chromatography to obtain material d;

[0031] D. The material d is subjected to the second high-performance liquid chromatography reversed-phase chromatography to obtain the target anti-gout active polypeptide RDP2.

[0032] The water described in the B step is deionized water.

[0033] The elution described in the B step is to take the Sephadex G50 column equilibrated with 20 mmol / L Tris-HCl buffer solution for 24 h in advance on the material b, and then use ...

Embodiment 1

[0040] Isolation, purification and identification of anti-hyperuricemia active polypeptide RDP2

[0041] 1. Separation and purification

[0042] The rice was soaked in deionized water overnight, and the supernatant was vacuum freeze-dried after filtration, and stored at -80°C for later use. The lyophilized powder of the obtained rice extract was dissolved in deionized water, and 1 mL of Sephadex G50 (GE Healthcare, ultra-fine) column (length 40 cm, inner diameter width 1.5 cm), eluted with the same buffer, the flow rate was 3 mL / 10 min, and collected once every 10 min. Detect the absorbance of each tube, the detection wavelength is 280 nm, the figure 1 The samples under the absorption peak indicated by the arrow were combined.

[0043] The obtained sample was loaded on a Hypersil ODS 25mm column (product of Yilite, with a size of 4.6 mm × 300 mm) equilibrated with ultrapure water (containing 0.1% trifluoroacetic acid) in advance, and the experimental instrument was a Waters...

Embodiment 2

[0048] Determination of Anti-gout Activity of Polypeptide RDP2

[0049] (1) Determination of serum uric acid content

[0050] The uric acid-lowering activity of polypeptide RDP2 was detected by oxonate potassium-induced hyperuricemia animal model. SPF grade Kunming mice with a body weight of 22-25 g were selected for the experiment, and the mice were randomly divided into 7 groups with 6 mice in each group. These 7 groups were blank control, model group, allopurinol group (positive control), benzbroma Long group (positive control) and three RDP2 treatment groups (100, 10, 5 µg / kg). The blank control group was given the same amount of normal saline by intragastric administration every day, and the other groups were given oxonate potassium (300 mg / kg) and adenine (200 mg / kg) by intragastric administration for 1 hour, and then intraperitoneally administered respectively: the model group was given etc. The positive control group was given 10 mg / kg allopurinol and 8 mg / kg benzbro...

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Abstract

The invention discloses an anti-gout active polypeptide RDP2 as well as a preparation method and application thereof. An amino acid sequence of the anti-gout active polypeptide RDP2 is AAAAGAMPK-NH2.The anti-gout active polypeptide RDP2 obtained by purification displays higher uric acid reducing activity in an oteracil potassium induced hyperuricemia animal model experiment and shows better relieving capacity in a monosodium urate crystal induced gout attack animal model, revealing that the anti-gout active polypeptide RDP2 has potential application prospects. The anti-gout active polypeptideRDP2 has the beneficial characteristics of simple structure and high anti-gout activity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an anti-gout active polypeptide RDP2 and its preparation method and application. Background technique [0002] With the development of the economy and the changes in the Chinese diet, gout has gradually evolved from a common occurrence in the West to a global occurrence. The 2017 China Gout Report White Paper shows that there are more than 80 million gout patients in China, and gout has become the second largest metabolic disease in China after diabetes. Gout is caused by the accumulation of uric acid in the blood and crystallization in joints or soft tissues. The pre-disease symptom is hyperuricemia. Gout not only induces severe pain during its acute attack period, affects people's quality of life, and leads to reduced joint function; but also during its occult period, that is, the non-attack period, persistent high uric acid can also increase the risk of cardio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/20A61K38/08A61P19/06
CPCC07K7/06A61P19/06A61K38/00
Inventor 杨新旺王滢唐璟孙俊刘乃心
Owner KUNMING MEDICAL UNIVERSITY
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