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TSG6 polypeptide fragments for dry eye disease

A dry eye disease and disease technology, applied in the field of treatment of dry eye disease, can solve problems such as changes in the stability of recombinant proteins, difficulties in large-scale production, etc.

Pending Publication Date: 2022-05-27
UNIV OF MANCHESTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, they concluded that the clinical application of TSG-6 is limited by various factors, including difficulties in large-scale production or changes in the stability of recombinant proteins

Method used

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  • TSG6 polypeptide fragments for dry eye disease
  • TSG6 polypeptide fragments for dry eye disease
  • TSG6 polypeptide fragments for dry eye disease

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0137] Example 1: LINK_TSG6 is more soluble than full-length TSG-6

[0138] The solubility and aggregation state of full-length TSG-6 and LINK_TSG6 in PBS were compared over a range of concentrations using UV spectrophotometry and dynamic light scattering (DLS). LINK_TSG6 was completely dissolved at 2 mg / ml and showed no aggregation at 0.4, 0.8, 1.6 or 3.2 mg / ml. The solubility of the full-length protein was much lower, with less than 40% of the protein remaining in solution at 0.4, 0.8, 1.6 and 3.2 mg / ml. The full-length protein was also highly aggregated at 0.2, 0.40.8, 1.6 and 3.2 mg / ml. DLS measurements of full-length TSG-6 at 1.6 and 3.2 mg / ml were out of range, indicating the formation of aggregates greater than 25 million Daltons.

example 2

[0139] Example 2: LINK TSG6 Ameliorates Signs and Symptoms of Dry Eye Disease in a Mouse Model

[0140] Topical TSG-6 has previously been shown to be as effective as cyclosporine eye drops in inflammation-mediated dry eye disease (Kim et al., 2016). To test whether the LINK_TSG6 polypeptide could be used for this indication, we investigated its effect in the NOD.B10 mouse model of primary Sjögren's syndrome (spontaneous dry eye without diabetes).

[0141] Put NOD.B10.H2 b Mice (12 weeks old, Jackson Lab), a model of primary Sjögren's syndrome (dry eye without diabetes), were treated with LINK_TSG6 topical application for 7 days. C57BL / 6 mice were used as a negative control as they did not suffer from spontaneous dry eye.

[0142] Mice were treated topically with 1 μg LINK_TSG6 in 10 μl PBS, 4 times a day (QID) for 7 days. Zolazepam-Teletamine by intraperitoneal injection Mice were anesthetized by Virbac, Carros, France and 10 μl of LINK_TSG6 or PBS was administered usin...

example 3

[0156] Example 3: The effect of LINK TSG6 on dry eye disease is dose-dependent

[0157] We were interested to know whether the effect observed in Example 1 was dose-dependent. To this end, we applied topically QID 1, 0.1 or 0.01 μg of LINK_TSG6 in 5 μl PBS to 12-week-old NOD.B10.H2b mice (Jackson Laboratory) for 7 days.

[0158] Mice were randomly assigned to the following treatment groups:

[0159] 1) Group 1 (negative control group): C57BL / 6 mice, 12 weeks old (n=6, 12 eyes) untreated

[0160] 2) Group 2 (positive control group): NOD.B10 (n=8, 8 eyes)+PBS 5μl QID

[0161] 3) Group 3 (experimental group): NOD.B10 (n=8, 8 eyes) + LINK_TSG6 1 μg (in 5 μl PBS) QID

[0162] 4) Group 4 (experimental group): NOD.B10 (n=8, 8 eyes)+LINK_TSG6 0.1μg QID

[0163] 5) Group 5 (experimental group): NOD.B10 (n=8, 8 eyes)+LINK_TSG6 0.01μg QID

[0164] The effect of the treatment was determined as in Example 1.

[0165] like Figure 3A Corneal epithelial defects were quantified in Li...

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Abstract

The present invention relates to the treatment of dry eye disease, and in particular, to (but not limited to) the treatment of dry eye disease with a LINKTSG6 polypeptide.

Description

[0001] This application claims priority to GB1910645.9 filed on July 25, 2019, the content and elements of which are incorporated herein by reference for all purposes. technical field [0002] The present invention relates to the treatment of ocular surface disorders, and in particular, but not limited to, the treatment of dry eye disease with LINK_TSG6 polypeptides. Background technique [0003] Tumor necrosis factor (TNF)-stimulated gene 6 (TSG-6) is the ~35 kDa secreted product of TNF-stimulated gene-6, expressed in response to inflammatory mediators and growth factors. [0004] Although constitutively expressed in a few tissues, TSG-6 is often upregulated where there is inflammation. For the most part, TSG-6 exhibits anti-inflammatory and tissue protective properties, but has been thought to sometimes play a role in disease pathology such as in the lungs. Although TSG-6 is produced by a wide range of cell types, TSG-6 was found to be produced by mesenchymal stem cells / s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P9/08A61P27/02A61P27/04A61K31/4725A61K31/573A61K38/13
CPCA61K38/1709A61K31/4725A61K31/573A61K38/13A61P27/04A61P27/02A61P9/08A61K9/0048A61K2300/00C07K14/4718A61P37/06
Inventor A·戴C·米尔纳吴周妍
Owner UNIV OF MANCHESTER
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