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Composition containing active prourokinase, freeze-drying process and freeze-dried preparation thereof

A technology of pro-urokinase and freeze-dried preparations, which is applied in the field of medicine and can solve problems such as instability of pro-urokinase preparations

Active Publication Date: 2008-09-24
TASLY BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of unstable pro-urokinase preparations in the prior art, the inventors of the present invention have obtained a composition containing active pro-urokinase through long-term research. Compared with the existing pro-urokinase preparations, the present invention The composition can be stored stably for a long time

Method used

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  • Composition containing active prourokinase, freeze-drying process and freeze-dried preparation thereof
  • Composition containing active prourokinase, freeze-drying process and freeze-dried preparation thereof
  • Composition containing active prourokinase, freeze-drying process and freeze-dried preparation thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Preparation of prourokinase

[0019] (See Chinese patent: preparation method of recombinant human glycosylated pro-urokinase, announcement number CN1062016C, announcement date: February 14, 2001)

[0020] (1) Cloning of prourokinase gene and construction of expression vector

[0021] Detroit 562 cells were induced by myristate (PMA), total RNA was extracted, and a cDNA library was constructed. A positive clone containing a gene fragment encoding urokinase was obtained by screening, and the full-length cDNA gene of human prourokinase was obtained. Gene recombination technology operation, the two transcription units expressing prourokinase and dihydrofolate reductase genes are placed in the same vector, which are controlled by metallothionein (MT) and SV40 early promoters respectively.

[0022] (2) Transfection and screening of highly expressed CHO engineered cells

[0023] Transfect 20-40 μg pMTSV-du plasmid DNA into CHO-dhfr by calcium phosphate coprecipitation method...

Embodiment 2

[0029] Vacuum Freeze Drying of Pro-uk Compositions

[0030] Purified Pro-uk pure product 2mg / ml, add mannitol to 6% (weight / volume ratio), albumin to 3‰ (weight / volume ratio), phosphate buffer concentration to 10mmol / L, chlorine The concentration of sodium chloride is 0.05mol / L, and it is divided into 1ml ampoules, put into a lyophilizer pre-cooled to -40°C, pre-frozen for 3 hours, and then vacuumized, and the temperature is raised to -30°C for the first time. After the internal moisture is basically sublimated, the temperature is raised for the second time to make the temperature rise to 30°C, and after stabilization, it is kept for 1 hour, and then the freeze-drying can be ended.

Embodiment 3

[0032] Vacuum Freeze Drying of Pro-uk Compositions

[0033] Purified Pro-uk pure product 4mg / ml, add mannitol to 6% (weight / volume ratio), albumin to 3‰ (weight / volume ratio), phosphate buffer concentration to 10mmol / L, chlorine The concentration of sodium chloride is 0.10mol / L, and it is divided into 1ml ampoules, placed in a freeze dryer that has been cooled to -40°C in advance, pre-frozen for 3 hours, and then vacuumized, and the temperature is raised to -30°C for the first time. After the internal moisture is basically sublimated, the temperature is raised for the second time to make the temperature rise to 30°C, and after stabilization, it is kept for 1 hour, and then the freeze-drying can be ended.

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Abstract

The invention belongs to the field of pharmacy, more specifically, the invention relates to a medicinal composition containing reactive pro-urokinase, its cryodesiccation method and freeze-dried preparation.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a pharmaceutical composition containing active prourokinase, a freeze-drying method and a freeze-dried preparation thereof. Background technique [0002] Cardiovascular system diseases caused by thrombus seriously threaten human health, and there is an urgent need for a drug with good thrombolytic effect and small side effects clinically. Pro-urokinase (Pro-uk) can specifically activate plasminogen on the surface of the thrombus, and then specifically dissolve the thrombus, while not destroying the fibrinolytic system, and the side effects of systemic bleeding are small; after preclinical animal experiments And clinical trials have proved that Pro-uk also has the advantages of small dosage, safe use, good thrombolytic effect, low re-thrombotic rate, and no allergic reaction. It is a safe, non-toxic, and ideal biological thrombolytic preparation. Broad market prospects. [0003] Since th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/49A61K9/19A61P7/02A61P9/10
CPCA61K38/49A61K9/19A61P7/02A61P9/10
Inventor 姜燕胡铁强肖成祖
Owner TASLY BIOPHARMACEUTICALS CO LTD
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