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New application of metallothionein mt-2 in the preparation of asthma therapeutic drugs

A technology of metallothionein and MT-2, which is applied in the field of protein-related drugs, can solve the problems of less research on the anti-asthma function of MT-2 protein, and there is no impact on lung compliance of tracheal smooth muscle cell systolic and diastolic functions, and achieve good clinical application Prospect, improvement of lung compliance, effects of inhibiting cell proliferation and migration

Active Publication Date: 2019-09-13
SHANGHAI UNIV OF T C M +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are few studies on the anti-asthma function of MT-2 protein, and there is no research on the effect of tracheal smooth muscle cell systolic and diastolic function, airway resistance, and lung compliance

Method used

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  • New application of metallothionein mt-2 in the preparation of asthma therapeutic drugs
  • New application of metallothionein mt-2 in the preparation of asthma therapeutic drugs
  • New application of metallothionein mt-2 in the preparation of asthma therapeutic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Preparation and purification of embodiment 1 metallothionein MT-2

[0043] According to the NCBI tissue distribution information, normal rat lung tissue was selected, total RNA was extracted by one-step method of guanidine isothiocyanate, and cDNA was synthesized by AMV reverse transcriptase, which was used as a template for PCR amplification of MT-2 gene under the action of DNA taq enzyme. PCR amplification, the upstream primer sequence of PCR amplification is 5'-AATGAATTCATGGACCCCAACTGCTCCT-3' (SEQ ID NO: 3), and the downstream primer sequence is 5'-AATCTCGAGGGCGCAGCAGCTGCACTT-3' (SEQ ID NO: 4). The PCR amplification program is as follows: the reaction system with a total amount of 20ul is pre-denatured at 94°C for 5min, then denatured at 94°C for 30s, annealed at 58°C for 30s, and extended at 72°C for 60s, for a total of 35 cycles, and finally extended at 72°C for 5min; The product sequence is shown in SEQ ID NO:2.

[0044] After the amplified product was digested w...

Embodiment 2

[0046] Embodiment 2 MT-2 protein influences on airway resistance and lung compliance of rat asthma model

[0047] 1. Experimental materials

[0048] 1.1 Experimental objects

[0049] SD rats, male, 4 weeks old, weighing 100-120 g.

[0050] 1.2 Experimental reagents

[0051] 1) The sensitization solution was injected intraperitoneally, which was physiological saline containing 1 mg OVA and 10 mg Al(OH)3, and 1 ml was injected intraperitoneally to each rat.

[0052] 2) The excitation solution is a normal saline solution containing 0.5% OVA, which is injected into the external jugular vein at a dose of 5 mg / kg.

[0053] 3) Other reagents are obtained by preparing analytically pure pharmaceutical saline.

[0054] 4) MT-2 protein, prepared in Example 1.

[0055] 5) Trx protein, as a protein negative control, the preparation method of this protein is as follows: transform pET32a plasmid into BL21(DE3) competent cells, culture at 37°C, 250rpm with shaking until OD 600 After abo...

Embodiment 3

[0085] Example 3 MT-2 Effector Protein Effects on Rat Isolated Tracheal Helical Strip Tension

[0086] 1. Experimental materials

[0087] 1.1 Experimental objects

[0088] Preparation of isolated rat bronchial smooth muscle spiral strips (bronchial spiral strips): After the rats were anesthetized with 1% pentobarbital sodium (50mg / kg), the trachea was separated and made into a strip with a width of about 5mm and a length of 15mm according to the method recorded in the literature. Helical strips, suspended in a gram-Heinz solution bath, filled with 95% O 2 +5%CO 2 gas to keep Po in solution 2 >53.3kPa, maintain a constant temperature of 37°C. The tracheal coil was connected to a tension transducer, and the tension changes were recorded with a physiological recorder. Before the experiment, the tracheal strips were equilibrated under the initial tension of 1.5g for 1h, and replaced with fresh Gram-Heinz solution every 20min. Use Ach (8mg / ml) to shrink the tracheal strip to ...

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Abstract

The invention relates to the field of protein drugs and particularly discloses an application of metallothionein MT-2 to preparation of an asthma therapeutic drug and a drug preparation containing the metallothionein MT-2. The invention discloses a new function of dilating the airway smooth muscle and reducing the airway resistance through the metallothionein MT-2. The airway smooth muscle can be obviously dilated, the airway resistance of an asthma model rat is reduced, the effect of curing the asthma is achieved, the metallothionein MT-2 can be used for preventing and curing the asthma disease and preparing the asthma therapeutic drug, and the application of the metallothionein MT-2 to curing of the asthma disease has the advantages of high safety, good effectiveness, high stability and the like.

Description

technical field [0001] The invention relates to the field of protein medicines, and specifically discloses the use of a metallothionein MT-2 for preparing asthma medicines and related asthma medicines. Background technique [0002] Metallothionein (MT) was first discovered in 1957 during the study of the biological effects of metals. It is called metallothionein because it contains rich sulfhydryl groups and can chelate a large amount of metal ions. Although the 20 members counted by the Metallothionein Nomenclature Committee generally have a low molecular weight and only contain 60-63 amino acids, their cysteine ​​content accounts for 23%-33%, and all of them are conservatively distributed. The metallothionein polypeptide chain does not contain aromatic amino acids, but each protein molecule can bind 7-12 metal ions through cysteine ​​sulfhydryl groups, thus having special spectral characteristics of metallothionein compounds. Mammalian metallothionein can be divided into ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61K38/17A61P11/06
Inventor 杨永清尹磊淼王宇徐玉东
Owner SHANGHAI UNIV OF T C M
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