Rabbit skin comprising biological active substances and its use

Abstract

A rabbit skin containing biologically active substances is obtained by the process including vaccinating rabbit skin tissues with vaccinia virus, feeding a rabbit vaccinated with vaccinia virus, killing the rabbit when its skin tissues are sufficiently inflamed, and skinning the rabbit. The rabbit skin of the present invention can be used for preparing drugs and health foods.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a rabbit skin containing biologically active substances and its use. BACKGROUND OF THE INVENTION [0002] It was reported that the extracts from inflammatory rabbit skin tissues vaccinated with vaccinia virus can be used for the treatment of allergic disease and have the analgesic effect. There has not been any method to prepare rabbit skin that contains strongly active and high yield biologically active substances.DETAILED DESCRIPTION OF THE INVENTION Aims [0003] The objective of the present invention is to provide a rabbit skin which is rich in biologically highly active substances that can be used for preparing drugs and health foods. Project [0004] As a result of many years of hard work, the inventors of the present invention have reached this aim. [0005] The rabbit skin of the present invention is obtained by the process including vaccinating rabbit (Oryctolagus cuniculus) skin tissues with vaccinia virus, feeding r...

AI Technical Summary

Benefits of technology

[0010] The rabbit skin of the present invention possesses 0.5 iu / g SART activity or more, and which also possesses the kallikrein-protease inhibition activity.

[0011] By extracting with organic solvent, processing with acid, processing with alkali, absorbing, eluting and concentrating, biologically active preparations being rich in amino acids and nucleic acids can be prepared from the rabbit skin. The said amino acids include glutamic acid, glycine, alanine, valine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, aspartic acid, threonine and serine. The said nucleic acids include urocanic acid, uracil, hypoxanthine, xanthine and thymine.

[0020] The rabbit skin was cut in pieces of 1 cm2, 4 times (w / w) of 3% phenol aqueous solution was added, then placed the mixture at 4° C. for 72 h, centrifuged after liquid changed into emulsion. The supernatant was filtrated to collect brown solution A. The brown solution A was boiled for 40 min in a water bath after pH was adjusted to 5.0 by 1M HCl, cooled to 28° C. promptly, centrifuged and filtrated to collect solution B. The solution B was boiled for 40 min in a water bath after pH of filtrate was adjusted to 9.2 by 1M NaOH, cooled to 28° C. promptly, and filtrated to collect solution C. pH of the solution C was adjusted to 4.5 by 1M HCl, activated charcoal was added at 30° C., stirred continuously for 4 h, stopped stirring, left it for 30 min, removed the supernatant, filtrated under nitrogen atmosphere. Then the activated charcoal was dipped in injectable water and washed, filtrated and discarded filtrate to collect the activated charcoal, reserved the activated charcoal. The activated charcoal was then put into injectable water, adjusted pH to 11.0 by 1M NaOH, stirred continuously for 4 h, filtrated with a 0.45-μm Millipore filter under nitrogen atmosphere, washed the activated charcoal by injectable water to collect solution D. After pH of the solution D was adjusted to 6.0 by 1M HCl, the vessel was sealed, heated up and kept the temperature at 121° C. for 20 min, and then cooled down to under 40° C. to collect solution E. The solution E was placed into a decompression distillator, replaced the air with nitrogen in the decompression distillator, distilled at 60° C. under decompression, filtrated to collect solution containing biologically active substances, whose SART activity was determined. The SART activity of the solution was adjusted to 1.2 iu / ml by evaporating, concentrating and diluting with distilled water. 10 ml of this solution was desalted at 10 μs / cm of final conductance, dried under decompression condition, into which 1.5 ml of 0.25 M NaCl solution was added to obtain test solution. 0.2 ml of 0.25M NaCl solution was regarded as the control solution and treated with parallel process with the 0.2 ml test solution. 0.5 ml human plasma were added respectively to both solutions, placed at freezing point for 5 min, added 0.25 ml of suspension of argilla, placed at freezing point for 20 min, and filtrated. 0.1 ml of filtrate was mixed with 0.2 ml of 0.1 M Tris-HCl buffer and 0.1 ml of basic solution. Effected reaction for 20 min at 30° C., and stopped the reaction by adding 0.8 ml of 1% citric acid. The absorbance A of test solution was determined in 405 nm as the absorbance of control solution was initialized as 0.4. If A was less than 0.4, the rabbit skin from which the test solution was prepared was regarded as possessing the kallikrein-protease inhibition activity.

Problems solved by technology

There has not been any method to prepare rabbit skin that contains strongly active and high yield biologically active substances.