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New preparation method of high-density lipoprotein

A high-density lipoprotein and apolipoprotein technology, which is applied in the application field of medicine, can solve the problems that limit the wide application of natural HDL injections, the scale is difficult to expand, and the burden on patients is increased.

Inactive Publication Date: 2010-12-29
吉林圣元科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0007] However, the only way to obtain HDL at this stage is to separate it from plasma. The source of raw materials is limited, and the production cost is high, and the scale is difficult to expand. The use of blood products is also easy to cause iatrogenic infection
These factors will increase the burden on patients and limit the wide application of natural HDL injections in clinical practice

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: the preparation of rhHDL

[0029] According to HDL 2 The ratio of various apolipoprotein components in the synthetic rhHDL, HDL 2 The ratio of various apolipoprotein components in the blood is as follows: ApoA-I 65%, ApoA-II 10%, ApoC-I 13%, ApoC-II 1%, ApoE 3% (Liu Ruijie, Niu Haomin, Wang Jingfu, etc. Hyperlipidemia syndrome and related diseases [M]. Beijing: Science and Technology Literature Publishing House, 1999).

[0030] 1) Take 7.2mg of lecithin and 1.6mg of cholesterol and dissolve them in 0.5ml of absolute ethanol. Inject quickly into 12.1ml 1 / 20PBS after inhalation with a skin test syringe, N 2 Stir for 15 min.

[0031] 2) 5.4 mg sodium cholate was dissolved in 0.5 ml 1 / 20 PBS; 3.2 mg rhApoA-I, 0.5 mg ApoA-II, 0.64 mg ApoC-I, 0.05 mg ApoC-II, 0.15 mg ApoE were dissolved in 0.8 ml 1 / 20 PBS. Add to the above liquid with stirring.

[0032] 3) Stand at room temperature for 30 minutes.

[0033] 4) Move to 4°C and incubate for 12h.

[0034] 5)...

Embodiment 2

[0037] Example 2: rhHDL mediates cholesterol efflux in rat peritoneal macrophages

[0038] 1) Establishment of experimental hyperlipidemia model

[0039] High-fat feed formulated with 1% cholesterol, 0.2% propylthiouracil, 0.5% sodium cholate, 2% lard, 5% soybean, 0.5% egg white, 0.5% fish meal, 2% egg yolk powder, and 88.3% basic feed Rats were fed for 15 days to establish a hyperlipidemia model.

[0040] 2) Induction of macrophages

[0041] Hyperlipidemic rats were intraperitoneally injected with 3% sterile sodium thioglycolate broth (2ml / rat) to induce the production of macrophages. Macrophages can be collected after 4 days.

[0042] 3) Isolation and culture of macrophages

[0043] Rats were killed by ether anesthesia, peritoneal macrophages were collected by peritoneal lavage with D-Hanks solution, centrifuged at 1000r / min for 10min, the precipitate was made into suspension with RPMI-1640 medium containing 10% calf serum, and the cell concentration was adjusted to 1×1...

Embodiment 3

[0055] Example 3: Effect of rhHDL on blood lipid metabolism in experimental hyperlipidemic rats

[0056] 1) Animal grouping and administration method

[0057] Fifty Wistar male rats, weighing 120-160 g, were randomly divided into 5 groups, 10 in each group. One of them was the blank control group, which was given normal feed for 15 days. The rest were in the high-fat diet group, given high-fat diet for 15 days, blood was collected on the 15th day to measure blood lipids, and then the high-fat diet group was randomly divided into 4 groups according to blood lipid levels, which were model control group, rhHDL low-dose group, rhHDL high-dose group, rhHDL high-dose group, positive control group. The grouping and administration methods are shown in Table 3.

[0058] Table 3 Animal grouping, administration method

[0059]

[0060] From the 16th day onwards, 8:00 to 9:00 in the morning of the next day, the tail vein was administered once, a total of 10 times. Animals were fas...

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Abstract

The invention relates to a new preparation method of high-density lipoprotein (rhHDL), in particular to a function of regulating blood fat metabolism through the rhHDL and application of the rhHDL to preparation of a medicament for preventing and treating blood fat metabolism abnormal diseases. The rhHDL is mainly characterized in that protein components comprise recombinant human apolipoprotein A-I, recombinant human apolipoprotein A-II, recombinant human apolipoprotein C-I, recombinant human apolipoprotein C-II and recombinant human apolipoprotein E which are prepared from methyl nutritional yeast; and the biological activity of the rhHDL is similar to that of natural high-density lipoprotein.

Description

field of invention [0001] The invention relates to the preparation of a new high-density lipoprotein (rhHDL), in particular to the function of rhHDL in regulating blood lipid metabolism, and its application in the production of medicines for preventing and treating abnormal blood lipid metabolism diseases. The main characteristic of rhHDL is that the protein components include recombinant human apolipoprotein A-I, recombinant human apolipoprotein A-II, recombinant human apolipoprotein C-I, recombinant human apolipoprotein C-II and recombinant human Apolipoprotein E 5 kinds of apolipoprotein; its biological activity is very similar to natural high-density lipoprotein. Background of the invention [0002] Metabolic syndrome patients and hyperlipidemia-induced coronary heart disease (coronary artery disease, CAD) patients all have obvious lipid metabolism disorders (elevated cholesterol and decreased HDL), and a large number of epidemiological studies have proved that plasma HD...

Claims

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Application Information

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IPC IPC(8): C12P21/02A61K38/17A61P3/06A61P9/10C12R1/84
Inventor 颜炜群苏曼曼
Owner 吉林圣元科技有限责任公司
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