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Medicine composition pancreatin enteric capsule and preparation method thereof

A technology of enteric-coated capsules and pancreatic enzymes, applied in the field of biomedicine, can solve problems such as disadvantages, no prompts, and inability to self-activate, and achieve the effects of good stability, cost saving, and low cost

Active Publication Date: 2013-11-20
浙江华润三九众益制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, Chinese patent 02805619 discloses a method for producing recombinant trypsin, which achieves secretory expression by expressing it in a pH3.0-4.0 system, and found that the truncated trypsin (pro) expressed more, but (self) activated The final activity is only 180-200U / mg, and there is no suggestion that the sequence changes at other sites will cause impact; Chinese patent 200910055493 increases the activity and stability of trypsin by adding a sequence at the N-terminus of the truncated trypsin (original) , its experimental data is worthy of verification, but at least its physiological temperature (37°C) cannot be auxiliary activated (not to mention self-activated), which is extremely unfavorable for administration in humans

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of trypsin

[0030] We have designed the optimal expression sequence of the trypsin gene of the present invention in yeast, its nucleotide is shown in SEQ ID NO: 1, and the amino acid sequence of the trypsin encoded by it is shown in SEQ ID NO: 2. We commissioned Shanghai Shenggong Company to artificially synthesize the gene sequence and clone it into yeast cells according to conventional methods to construct expression engineering bacteria. In short, using the synthesized gene sequence as a template, using the The upstream and downstream primers were amplified by PCR, and then connected between EcoR I and Xba I of pPICZαA (available from Invitrogen, USA), and the positive construct was electrotransformed into yeast GS115 strain (available from Invitrogen, USA), and screened with Zeocin The positive yeast cells were used as engineering bacteria.

[0031] Inoculate the constructed engineered bacteria into 25ml seed culture medium (formulation: 2% ...

Embodiment 2

[0033] Example 2 Study on properties of trypsin

[0034]The trypsin prepared in Example 1 was dissolved in pH 8.0 Tris-HCl buffer solution containing 20 mM calcium chloride, put into a dialysis bag, and dialyzed twice in pH 8.0 Tris-HCl buffer solution (0.1M) at 4°C. Each time, 15 minutes each time, then adding Tos-Gly-Pro-Arg-4-pNA (Chromozym TRY) as a substrate, autoactivating trypsin at different temperatures for 1 hour, and then detecting its activity. The results are shown in Table 1. The trypsin of the present invention has high self-activation at physiological temperature, but almost no activity at low temperature, and the enzyme activity at 37°C can reach more than 300U / mg in a short time.

[0035] Table 1 Properties of pancreatin of the present invention

[0036] temperature (°C) 4 25 37 50 Activity (U / mg) 2.8 276 311 65

Embodiment 3

[0037] The preparation of embodiment 3 trypsin enteric-coated capsules

[0038] (1) Dissolve hydroxypropylmethylcellulose in water, then add 95% ethanol solution to prepare 2% (W / W) hydroxypropylmethylcellulose in ethanol aqueous solution (ethanol: water (volume ratio) = 1: 1).

[0039] (2) Mix 28 parts of pancreatin, 8 parts of sucrose and 4 parts of dextrin prepared in Example 1 and put them in a centrifugal granulator (available from Shandong Linyi Huanshi Micropill Equipment Co., Ltd., BZL-300 type) According to the manufacturer's instructions, spray 32 parts of the ethanol aqueous solution of hydroxypropyl methylcellulose in step (1) (it has been measured that about 0.6 parts of hydroxypropylmethylcellulose is bound to the core of the pellet), and granulate to form The ball core, after drying, has an average particle size of 0.9mm and is ready for use.

[0040] (3) Add 3 parts of triethyl citrate to 500 parts of distilled water, mix well and then transfer into ...

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PUM

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Abstract

The invention provides pancreatin and an enteric capsule thereof. An amino acid sequence of the pancreatin is shown in SEQIDNO:2 and the like; a particle in the pancreatin enteric capsule is composed of a pill core and a coating wrapped outside the pill core, wherein the pill core is composed of the pancreatin, cane sugar, dextrin and hydroxypropylated methylcellulose, and the coating is composed of polyacrylic resin, triethyl citrate and talcum powder.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular, the invention relates to a pancreatin and an enteric-coated capsule thereof. In addition, the present invention also relates to the preparation method and application of the above product. Background technique [0002] Pancreatin, synthesized in mammalian pancreatic cells as trypsinogen, is activated by enterokinase and is also self-activating at mildly alkaline pH (eg, physiological pH). For example, Chinese patent 02805619 discloses a method for producing recombinant trypsin, which achieves secretory expression by expressing it in a pH3.0-4.0 system, and found that the truncated trypsin (pro) expressed more, but (self) activated The final activity is only 180-200U / mg, and there is no suggestion that the sequence changes at other sites will cause impact; Chinese patent 200910055493 increases the activity and stability of trypsin by adding a sequence at the N-terminus of the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/94C12N15/52A61K9/48A61K38/43A61P1/00A61P1/14
Inventor 金鑫朱菊红毛道斌丁莉梅傅姗瑚
Owner 浙江华润三九众益制药有限公司
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