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Method capable of reducing immunotoxicity of venom nerve growth factor and enabling immunotoxicity to be drug alternative of mouse origin nerve growth factor

A nerve growth factor and drug technology, applied in the field of nerve growth factor-specific binding of small lipid molecules, can solve problems such as unclear mechanism

Active Publication Date: 2015-01-07
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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Problems solved by technology

For example, the combined action of NGF extracted from snake venom and rat submandibular gland and lysoPS can induce the degranulation reaction of mast cells and release a variety of inflammatory mediators: such as 5-hydroxytryptamine (5-HT), histamine (hiatamine) and interleukins, etc. , but its mechanism is still unclear

Method used

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  • Method capable of reducing immunotoxicity of venom nerve growth factor and enabling immunotoxicity to be drug alternative of mouse origin nerve growth factor
  • Method capable of reducing immunotoxicity of venom nerve growth factor and enabling immunotoxicity to be drug alternative of mouse origin nerve growth factor
  • Method capable of reducing immunotoxicity of venom nerve growth factor and enabling immunotoxicity to be drug alternative of mouse origin nerve growth factor

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Embodiment 1

[0032] The invention provides a purification method and a degreasing method of snake venom nerve growth factor cNGF (SEQ ID NO: 1). Histamine release experiment of cNGF and defatted cNGF in mouse mast cell tumor cells.

[0033] 1. Natural extraction and purification of Chinese cobra venom nerve growth factor NGF protein.

[0034] The purification scheme is: first step ion column + second step ion column + molecular sieve

[0035]The crude snake venom we purchased from the Snake Venom Research Institute of Guangxi Medical University comes from Chinese cobra (N.naja atra). Those skilled in the art should understand that, in addition to the crude venom of the Chinese cobra, other commercially available crude venoms can also be used. The present invention has no special limitation on the species source of the crude snake venom.

[0036] The crude snake venom was dissolved in 0.1M sodium acetate solution at pH 4.6 with a concentration of 100mg / ml. After the fully dissolved crud...

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Abstract

The invention discovers a cobra nerve growth factor (cNGF) specific binding lipid small molecules comes from a cobra is a main source of the immunotoxicity of the cNGF according to studies of structural biology and biochemistry. Therefore, the invention relates to a purification method and a degreasing method of the cobra venom cNGF. Due to the fact that the purified cNGF is processed by means of organic solvent and a special process, the lipid small molecules are successfully eliminated, the immunotoxicity of mouse mastocytoma is reduced by means of the degreased cNGF according to confirmation of a cell experiment, and therefore the immunotoxicity of the cNGF can be used as the drug alternative of the mouse origin nerve growth factor with low side effects and is a drug which has a neurotrophic effect.

Description

Field of invention: [0001] The present invention finds that the nerve growth factor (cNGF) from the cobra specifically binds small lipid molecules through structural biology and biochemical studies, and these small lipid molecules are one of the main sources of its immunotoxicity. Therefore, the present invention provides a purification method and degreasing method of cobra venom nerve growth factor (cNGF), utilizes an organic solvent to take a special process to degrease the purified cNGF, successfully removes small lipid molecules, and confirms by cell experiments The delipidated cNGF reduces the immunotoxicity to mouse mast cell tumor cells, and can be used as a substitute drug for mouse nerve growth factor as a drug with low side effects and neurotrophic effect. Background technique: [0002] Nerve growth factor (NGF) is widely distributed in the human body and is a biologically active protein with complex functions. Mature biologically active NGF exists in organisms in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/48C07K1/18C07K1/16A61K38/18A61P25/00
Inventor 江涛唐捷童琼周洪哲孙含丽
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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