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Method and compsn. of nervous regulation protein for treating myocardial infarction

A technology of neuregulin and myocardial infarction, applied in drug combination, peptide/protein composition, pharmaceutical formula, etc., can solve the problem that it is difficult to observe the effect of Neuregulin-1

Inactive Publication Date: 2004-05-26
ZENSUN (SHANGHAI) SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, in the aforementioned Neuregulin-1 gene inactivation experiments, it was difficult to observe the role of Neuregulin-1 in the adult heart due to the early death of embryos

Method used

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  • Method and compsn. of nervous regulation protein for treating myocardial infarction
  • Method and compsn. of nervous regulation protein for treating myocardial infarction
  • Method and compsn. of nervous regulation protein for treating myocardial infarction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Amplification of the neuregulin gene

[0072] The neuregulin gene is located on chromosome 8P12 and has about 13 exons. The constructed neuregulin consists of 61 amino acids, the theoretical molecular weight is 7055D, the apparent molecular weight of SDS-PAGE electrophoresis is 6500-7000D, the isoelectric point is around 6.5, there is no glycosylation site, and it contains three disulfide bonds. Expression was performed using the E. coli system. PET22b was selected as the expression plasmid, and the human neuregulin gene was introduced to transform Escherichia coli BL21. After screening, an engineering strain with high expression of recombinant human neuregulin was obtained.

[0073] Amplification of the neuregulin gene. Total RNA and mRNA were extracted from human 5-month-old embryonic brain tissue, reverse-transcribed into cDNA, and used as a template for RT-PCR to obtain the target gene. The PCR amplification product was subjected to 1.5% agarose electr...

Embodiment 2

[0074] Embodiment 2 Preparation of recombinant human neuregulin

[0075] The human neuregulin gene was cloned into the PET22b expression plasmid by the calcium chloride precipitation method, and a recombinant human neuregulin expression plasmid (PET22b-human neuregulin) was constructed. This protein was highly expressed under the drive of the T7 promoter. The N-terminal of the expressed gene is inserted from the NdeI site, and the C-terminal has a terminator next to the last amino acid, and the resulting protein does not form a fusion protein with any amino acid. The correct fragment of about 183bp was excised by enzyme digestion. After the transformants were identified by enzyme digestion, the double-stranded DNA was extracted for nucleotide sequence determination. The result confirmed that the human neuregulin sequence on the expression vector was completely correct.

[0076] The sequence of neuregulin cDNA is (SEQ ID NO: 2):

[0077] AGC CAT CTT GTA AAA TGT GCG GAG AAG G...

Embodiment 3

[0085] Example 3. NRG-1 Therapeutic Experiments on Animals with Myocardial Infarction

[0086] Experimental materials and methods

[0087] Animals: 300-350 g, male, SD rats (provided by the Experimental Animal Center of Shanghai Medical University) were selected.

[0088] Model making method: according to Johns and Olson (Johns, T.N.P., and B.J.Olson. Experimental myocardial infarction. I.A method of coronary occlusion in small animals. Ann. Surg. 140: 675-682, 1954.) Ligate the left coronary artery of rats, ligate One week later, the echocardiogram was performed, and rats with a EF (ejection fraction) drop of about 70% were selected as successful myocardial infarction models.

[0089] Grouping and treatment: divided into 4 groups, 10 in each group. Sham operation group: perform surgery but do not ligate the left coronary artery of the rats, without any treatment; negative control group: inject PBS into the tail of myocardial infarction rats; positive control group: intragas...

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Abstract

An application of the neuregulin, its functional fragment, the nucleic acid for coding said neuregulin or its functional, fragment, or the substances for increasing the output and function of said neuregulin in preventing, treating and relaxing the myocardial infarction is disclosed. Its components, reagent kit and composition are also disclosed.

Description

Technical field: [0001] The present invention relates to components and methods for preventing, treating or delaying myocardial infarction in mammals, especially humans. More specifically, the present invention provides a method for preventing, treating or delaying myocardial infarction in mammals, the method comprising administering an effective amount of neuregulin, or its Functional fragments, or nucleic acids encoding the protein or functional fragments, or substances that increase the yield and (or) function of the neuregulin, so as to prevent, treat or delay the myocardial infarction. The present invention also provides neuregulin and substances for improving the yield and (or) function of neuregulin for preventing, treating or delaying myocardial infarction in mammals, especially human beings, kits and effective pharmaceutical ingredients. technical background: [0002] Myocardial infarction is coronary artery occlusion, blood flow interruption, causing partial necro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P9/10
Inventor 周明东
Owner ZENSUN (SHANGHAI) SCI & TECH CO LTD
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