Chimeric polypeptide with dual-targeting function and applications thereof

A chimeric polypeptide and dual-targeting technology, which is applied in the preparation of protein-based polypeptides and its application field, to achieve the effects of increasing expression, strong pertinence, and reducing the number of medication

Inactive Publication Date: 2010-04-28
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using the enzymes present in the human body to crack the polypeptide drug in the body and slowly release its active ingredients, making it a polype

Method used

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  • Chimeric polypeptide with dual-targeting function and applications thereof
  • Chimeric polypeptide with dual-targeting function and applications thereof
  • Chimeric polypeptide with dual-targeting function and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Preparation of chimeric polypeptide of the present invention

[0030] according to figure 1 According to the arrangement design, a chimeric peptide sequence composed of 80 amino acids at the N-terminal of lysozyme, 2 amino acid linking sequences and Exendin-4 was generated by genetic engineering technology, the sequence is as follows:

[0031] MKALIVLGLVLLSVTVQGKVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNT

[0032] RATNYNAGDRSTDYGIFQINSRGPHGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS (SEQ ID No: 3)

[0033] The underline is the connection sequence, and P in the connection sequence can also be A, S, V, L.

[0034] The specific steps for preparing the above sequence are as follows:

[0035] 1. Obtain the DNA sequence of 80 amino acids at the N-terminal of human lysozyme and the cleavage sites of thrombin and dipeptidase by using PCR method or chemical synthesis method;

[0036] 2. Obtain the linking sequence containing thrombin and dipeptidase cleavage site and the DNA sequen...

Embodiment 2

[0048] according to figure 1 According to the arrangement design, a chimeric polypeptide consisting of 60 amino acids at the N-terminal of lysozyme, 6 amino acid linking sequences and Exendin-4 was synthesized by chemical synthesis, and then purified and prepared according to conventional methods. Peptide pharmaceutical ingredients.

[0049] The sequence of the chimeric polypeptide in this example is:

[0050] MKALIVLGLVLLSVTVQGKVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNT

[0051] RALVPRGPHGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS (SEQ ID No: 4)

[0052] The underline is the connection sequence, L and V in the connection sequence can also be A, I, M, F, P, W, etc., and the P in the last position of G in the connection sequence can also be A, S, V, L.

Embodiment 3

[0054] according to figure 1 According to the arrangement design, a chimeric polypeptide composed of 40 amino acids at the N-terminal of lysozyme, 12 amino acid linking sequences, and Exendin-4 was synthesized by chemical synthesis, and then purified and prepared according to conventional methods for this chimeric polypeptide drug suitable for human Element.

[0055] The sequence of the chimeric polypeptide in this example is:

[0056] MKALIVLGLVLLSVTVQGKVFERCELARTLKRLGMDGYRGLVPRGPLVPRGPHGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS (SEQ ID No: 5)

[0057] The underline is the connection sequence, L and V in the connection sequence can also be A, I, M, F, P, W, etc., and the P in the last position of G in the connection sequence can also be A, S, V, L.

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Abstract

The invention provides a chimeric polypeptide with dual-targeting function, and a preparation method and applications thereof. The chimeric polypeptide of the invention comprises three sequences with the following characteristics: sequence 1 is an amino acid sequence comprising the first 20-80 human lysozyme proteins counting from end N; sequence 2 is an amino acid sequence of Exendin-4 proteins; and sequence 3 is a connecting sequence of the sequence 1 and the sequence 2 and comprises a thrombin restriction enzyme site and a dipeptidyl peptidase restriction enzyme site. The chimeric polypeptide can be synthesized by using a polypeptide synthesizing technology, or can be obtained by constructing an expression carrier comprising the polypeptide nucleotide sequence specified by the code, expressing in a genetic engineering stain and finally purifying. The chimeric polypeptide of the invention has obvious dual-targeting activity for resisting AGEs and reducing blood sugar, and can be used for preparing medicines for resisting AGEs and/or lowering high blood sugar.

Description

technical field [0001] The invention relates to the field of preparation and application of protein-made polypeptides, in particular to a dual-targeting chimeric polypeptide comprising human lysozyme N-terminal amino acid sequence and Exendin-4 amino acid sequence and its application. Background technique [0002] Lysozyme is a defensive enzyme present in egg white and body secretions (such as saliva, tears, etc.), which can effectively hydrolyze bacterial wall polysaccharides. Human lysozyme is composed of 148 amino acid residues, and its structure is shown in SEQ ID No: 1, in which there is a domain composed of about 18 amino acid residues near the N-terminal and has a high degree of advanced glycation end products (AGEs) Affinity, its dissociation constant Kd is 50nM. In vitro experiments found that lysozyme can bind AGEs-modified albumin and promote the uptake of AGEs by monocyte-macrophages. Based on the fact that AGEs in the body are degraded by extracellular enzymes...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K1/00C12N15/62C12N15/63A61K38/17A61P3/10
Inventor 刘誉谢琪璇
Owner JINAN UNIVERSITY
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