Matrix metalloproteinases and uses thereof

A matrix metal, protease technology, applied in the direction of peptide/protein components, enzymes, peptidases, etc., can solve problems such as instability

Inactive Publication Date: 2016-01-13
克劳迪娅·齐尔贝尔博格 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

An obvious problem is that collagenase I is the least stable component to release enzyme HI, as form Ia is rapidly autocatalytically degraded to form Ib

Method used

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  • Matrix metalloproteinases and uses thereof
  • Matrix metalloproteinases and uses thereof
  • Matrix metalloproteinases and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0262] Example 1: MMP-induced isolation of stem cells from adipose tissue

[0263] Isolation of adipose-derived stromal stem cells (ADSCs) has mainly been achieved with collagenase I or liberase consisting of collagenases I and II and thermolysin (Priya, Sarcar et al., (2012) J. Tissue Eng. Regen . Med. Jul. 27:1-9). Collagenase types I and II were purified from the highly dangerous Clostridium spp., an agent of gas gangrene. Furthermore, crude preparations from Clostridium histolyticum contained not only some collagenases, but also thiol proteases, clostripain, trypsin-like enzymes and aminopeptidases. During the release of the enzyme production, the collagenase isoenzyme is purified by a process of removing the considerable amount of endotoxin present in the raw material. There was a wide range of endotoxin contamination of conventional collagenase preparations compared to endotoxin levels of releasing enzymes. However, regardless of the source, all purified collagenases ...

Embodiment 2

[0288] Example 2: Separation of various cells

[0289] Islet isolation from pancreas: There are three main steps: in situ perfusion of the pancreas with an MMP-mixture (eg two or more MMPs), digestion of the pancreas and purification of the islets.

[0290] Mice are first anesthetized with pentobarbital or the like and then moved into a hood. Each mouse was laid down, belly side up, and its skin was disinfected with 70% ethanol. An incision is made around the upper abdomen to expose the liver and intestines. Clamp the ampulla of the mouse against the duodenal wall with surgical forceps to block the path of bile to the duodenum. Prepare 3 ml of buffer by dissolving the MMP mixture in 5 ml of 1X HBSS and draw into a 5 ml syringe fitted with a 30G1 / 2-G needle. The needle is then inserted under a microscope into the common bile duct through the junction of the hepatic and cystic ducts and into the middle of the common bile duct. The solution is injected slowly to expand the panc...

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Abstract

Matrix metalloproteinases (MMPs) compositions, inactive forms of MMPs (e.g. proMMPs), fragments, mutants, variants or combinations thereof. A pharmaceutical composition comprises one or more of the above in a pharmaceutical carrier. A composition comprises at least one of: a matrix metalloproteinase (MMP), an inactive MMP or a proenzyme (proMMP) thereof, wherein the matrix metalloproteinase (MMPs), inactive MMPs or proMMPs thereof, comprise: MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13.MMP-14, MMP-15, MMP-16, MMP-17, MMP-18, MMP-19, MMP-20, MMP-21, MMP-23A, MMP-23B, MMP-24, MMP-25, MMP-26, MMP-27, MMP-28, active fragments, mutants, variants or any combinations thereof. The uses include isolation of cells, in particular stem cells, from tissues, dissociation of tissues, proteins and treatment of a variety of conditions.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application Serial No. 61 / 808,861, entitled "Matrix Metalloproteases and Uses Thereof," filed April 5, 2013, the entire contents of which are incorporated herein by reference. technical field [0003] Embodiments relate to compositions, methods of use, and therapeutic applications of matrix metalloproteinases (MMPs). Background technique [0004] Mesenchymal stem cells (MSCs) are known to have the ability to self-renew and differentiate into cell types of the mesenchymal lineage including osteoblasts, adipocytes, chondrocytes, tenocytes, and myoblasts, and contribute to various mesenchymal Tissue regeneration. MSCs can be obtained after a bone marrow aspiration (aspiration) procedure and then cultured in vitro without loss of their stem cell potential, making them attractive targets for cell therapy. However, the traditional bone marrow harvesting procedure is pai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/08A61K38/16C12N9/50C12N9/64
CPCA61K38/4886C12N9/6416C12N9/6491C12Y304/24C12Y304/24007C12Y304/24017C12Y304/24024C12Y304/24034C12Y304/24035C12Y304/24065A61P3/04A61P43/00
Inventor 克劳迪娅·齐尔贝尔博格格雷格·B·菲尔茨
Owner 克劳迪娅·齐尔贝尔博格
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