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Modified growth differentiation factor and preparation and application methods thereof

A growth differentiation factor and differentiation factor technology, applied in the biological field, can solve the problems of low biological activity and short plasma half-life, and achieve the effects of simple modification process, increased half-life, and improved anti-apoptotic activity.

Active Publication Date: 2019-03-19
广东美赛尔细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] Aiming at the problem that the plasma half-life of growth differentiation factor 11 (GDF11) is relatively short and its biological activity is low, the purpose of the present invention is to provide a modified GDF11

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  • Modified growth differentiation factor and preparation and application methods thereof
  • Modified growth differentiation factor and preparation and application methods thereof
  • Modified growth differentiation factor and preparation and application methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1 A kind of modified growth differentiation factor

[0052] The modified growth and differentiation factor is obtained by specifically modifying the N-terminus of the natural growth and differentiation factor 11 protein derived from mammals by methoxy polyethylene glycol-propionaldehyde (mPEG-ALD).

[0053] The preparation method of the modified growth differentiation factor (PEG-rhGDF11) is:

[0054] S1. Take the growth differentiation factor 11 solution with a concentration of 1 mg / mL in a phosphate buffer solution with a pH value of 7.2 and a concentration of 50 mmol / L, and stir at 300 / min for 5 min until completely dissolved to obtain a mixture I;

[0055] S2. Add mPEG-ALD of the same mass as growth differentiation factor 11 to the mixture I obtained in step S1, react at 4°C for 10 h, add glycine to terminate the reaction, and obtain a crude reactant;

[0056] S3, using Superdex 200 molecular sieves for the crude reactant obtained in step S2, with a conc...

Embodiment 2

[0058] Embodiment 2 A kind of modified growth differentiation factor

[0059] The modified growth and differentiation factor is obtained by specifically modifying the N-terminus of the natural growth and differentiation factor 11 protein derived from mammals by methoxy polyethylene glycol-propionaldehyde (mPEG-ALD).

[0060] The preparation method of the modified growth differentiation factor (PEG-rhGDF11) is:

[0061] S1. Take the growth differentiation factor 11 solution with a concentration of 1 mg / mL in a phosphate buffer solution with a pH value of 7.6 and a concentration of 50 mmol / L, and stir at 500 r / min for 10 min until completely dissolved to obtain a mixture I;

[0062] S2. Add mPEG-ALD of the same mass as growth differentiation factor 11 to the mixture I obtained in step S1, react at 4°C for 15 h, add glycine to terminate the reaction, and obtain a crude reactant;

[0063] S3, using Superdex 200 molecular sieves for the crude reactant obtained in step S2, with a c...

Embodiment 3

[0065] Embodiment 3 A kind of modified growth differentiation factor

[0066] The modified growth and differentiation factor is obtained by specifically modifying the N-terminus of the natural growth and differentiation factor 11 protein derived from mammals by methoxy polyethylene glycol-propionaldehyde (mPEG-ALD).

[0067] The preparation method of the modified growth differentiation factor (PEG-rhGDF11) is:

[0068] S1. Take the growth differentiation factor 11 solution with a concentration of 1 mg / mL in a phosphate buffer solution with a pH value of 7.4 and a concentration of 50 mmol / L, and stir at 400 r / min for 8 min until completely dissolved to obtain a mixture I;

[0069] S2. Add polyethylene glycol active ester of the same mass as growth differentiation factor 11 to the mixture I obtained in step S1, react at 4° C. for 13 h, add glycine to terminate the reaction, and obtain a crude reactant;

[0070] S3, using Superdex 200 molecular sieves for the crude reactant obta...

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Abstract

The invention belongs to the field of biotechnology and particularly relates to a modified growth differentiation factor and preparation and application methods thereof. The modified growth differentiation factor is prepared by taking methoxy-polyethylene glycol-propionaldehyde as a modifier to perform specific modification on the N terminal of growth differentiation factor 11 protein, performingmodification reaction, separation and purification and collecting eluent. The growth modification factors modified by methoxy-polyethylene glycol-propionaldehyde can be applied to resisting ageing andimproving the cardiocerebral vascular system, is high in stability and significantly increased in half-life period and improves the inhibiting effects on apoptosis and the protective effects on endotheliocytes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a modified growth differentiation factor and its preparation method and application. Background technique [0002] Growth differentiation factor 11 (GDF11), also known as bone morphogenetic protein 11 (BMP11), belongs to the transforming growth factor-β (Transforming growth factor-β, TGF-β) superfamily. A new growth differentiation factor was first reported in 1999. In early embryonic development, GDF11 participates in the formation and differentiation of tissues and organs including bone, kidney, pancreas, retina, and olfactory nerve through negative regulation, and is an indispensable molecule for normal embryonic development. In recent years, studies have found that GDF11 can significantly improve brain cognition, reverse cardiac hypertrophy, improve cerebrovascular system, improve skeletal muscle metabolism and other functions, showing the extensive biological activi...

Claims

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Application Information

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IPC IPC(8): C07K14/51C07K1/107C07K1/16A61K38/18A61K47/60A61P25/00A61P9/00A61P9/10A61P19/08A61P3/10A61P3/06A61P9/14
CPCA61K38/1875A61K47/60A61P3/06A61P3/10A61P9/00A61P9/10A61P9/14A61P19/08A61P25/00C07K14/51
Inventor 任哲刘俊伟廖晓凤梅建勋
Owner 广东美赛尔细胞生物科技有限公司
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