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Hatching fluid enzymes and uses thereof

A technology of sequences and amino acids, applied in the field of hatching liquid enzymes and their uses, can solve the problem of high price

Active Publication Date: 2015-03-25
AQUA BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] The biggest disadvantage of exfoliation is the high price of some of the products and methods used to achieve said exfoliation

Method used

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  • Hatching fluid enzymes and uses thereof
  • Hatching fluid enzymes and uses thereof
  • Hatching fluid enzymes and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] Example 1: Identification and Characterization of VAPs

[0184] protein separation

[0185] During the analysis of the hatching fluid composition of Atlantic salmon, new proteins were identified that were present in the hatching fluid.

[0186] A method for preparing a portion of the hatching fluid from which zonase can be prepared which can be used as starting material for isolating the VAPs (or their precursor sequences) of the invention is provided in WO99 / 29836, incorporated herein For reference (in particular Example 1 of the process, but optionally without the urea step).

[0187] Therefore, the following method was used for separation. VAPs were isolated from hatching fluid (crude or filtered through a 0.45 μm filter). VAPs were then precipitated by adding 4x volumes of acetone at room temperature or 4°C. After 20-30 minutes, the precipitated VAPs were collected as a pellet after centrifugation at low speed (about 5000 xg) and resuspended in a suitable buff...

Embodiment 2

[0196] Example 2: In vitro medical / cosmetic applications of VAPs

[0197] Materials and methods

[0198] The following studies were performed with Atlantic salmon VAPs prepared as described in Example 1.

[0199] Differentiated human skin epithelial cultures were obtained from SkinEthics (Nice, France) 16 days after seeding onto plastic growth substrates with micropores allowing nutrients to reach the epithelial tissue from below. Such cultures exhibit normal skin morphology after differentiation during incubation at 37°C. These cultures were maintained in vitro for an additional 2 days, exposing the upper stratum corneum to air and the basal layer to the growth medium.

[0200] Parallel cultures were moved to media provided with a phosphate-buffered saline matrix containing Ca, Mg in the presence or absence of 0.5 mg / ml VAPs (measured at OD280) in a humidified atmosphere at 30°C for 6 hours. Cultures were formalin fixed and paraffin embedded and stained with hematoxylin / ...

Embodiment 3

[0205] Example 3: Medical / cosmetic application of choriolysin L in vivo

[0206] Materials and methods

[0207] The following studies were performed using choriolysin L of Atlantic salmon prepared from salmon hatching fluid as described by Yasumasu et al. 1989, supra.

[0208] Human skin epithelial cultures were prepared as described in Example 2, and 0.15 mU / ml oocysin L from salmon hatchery fluid was applied for 6 hours at 30°C.

[0209] result

[0210] A. Exfoliation

[0211] Figure 3A Results are shown in and B, where A shows skin cultures exposed to salmon ooshell lysin L and B shows control skin cultures. The results showed that choriolysin L caused the skin layers to delaminate and crack.

[0212] Exfoliation can also be analyzed by assessing the supernatant of the skin culture to assess the amount of epithelial cells removed from the skin culture during treatment. Since chotolysin L is inhibited by 1 mM EDTA, its effect can be easily inhibited to demonstrate i...

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Abstract

The present invention relates to various polypeptides from fish hatching fluid, their encoding nucleic acid sequences, pharmaceutical compositions comprising said polypeptides and nucleic acid molecules and their use in various medical and cosmetic applications to the skin, particularly for moisturizing skin and / or for exfoliation of the horny layer of the skin for treating or preventing skin disorders or conditions in an animal.

Description

[0001] This application is a divisional case of the international application date of November 30, 2010, the international application number PCT / EP2010 / 068509 entering the Chinese national phase on May 30, 2012, the application number 201080054198.1, and the invention name "hatch liquid enzyme and its use" Application. technical field [0002] The present invention relates to the use of choriolysin and strongly acidic protein (VAP) from fish hatching fluid alone or in combination in various cosmetic and medical applications for the skin. The invention also relates to strongly acidic proteins described for these uses. Background technique [0003] The skin is one of the more delicate organs of the human body. Although rarely life-threatening, skin diseases or conditions can be uncomfortable and potentially chronically incapacitating. Additionally, skin diseases and conditions can lead to psychological stress because the skin is clearly visible. Accordingly, there remains ...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K31/7088A61K8/64A61K8/60A61Q19/00A61P17/16A61P17/12A61P17/00A61P17/10A61P17/06C07K14/46C12N15/12
CPCA61K2800/28A61Q19/00A61K8/64C12N9/6402A61Q19/10A61Q19/02A61K8/606C07K14/461A61K38/00C12N9/6416A61K38/1706A61K38/4886C12Y304/24066A61P17/00A61P17/04A61P17/06A61P17/10A61P17/12A61P17/16A61K9/0014A61K9/06C07K14/46A61K8/66C12N9/64A61Q19/007
Inventor 汉斯·克里斯蒂安·莱伦贝恩特·沃瑟尔
Owner AQUA BIO TECH
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