Cellular glycosaminoglycan compositions and methods of making and using
Inventors
GLASS, Charles • THACKER, Bryan • ESKO, Jeffrey
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11549000-B2
Publication Date
2023-01-10
Expiration Date
Abstract
Disclosed herein are substantially pure glycosaminoglycan compositions derived from genetically modified cells.
Core Innovation
The invention relates to glycosaminoglycan compositions, including heparan sulfate and chondroitin sulfate, with defined sulfation or defined modification patterns. The compositions are substantially pure, genetically engineered-cell-derived glycosaminoglycan (GAG) compositions comprising heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, and hyaluronic acid, and are substantially free of contaminating GAG types.
Genetically modified cell line sources are used to generate defined GAG sulfation and other modifications through genetic modifications of glycosyltransferases, sulfotransferases, epimerases, sulfatases, and related targets. The engineered modifications include deficiencies and transgenes intended to produce defined patterns of sulfation and epimerization of uronic acid, and the resulting heparan sulfate can have reduced or increased binding and/or signaling with respect to FGF2, relative to heparan sulfate derived from a wild-type CHO cell.
The compositions are supported by analytical characterization directed to validating the defined GAG identity and modification state, including CRISPR/Cas mutation assays, flow cytometry/antibody detection of CS/HS on cell surfaces, GAG quantification, and disaccharide LC/MS analysis. The invention also includes a pharmaceutical composition comprising the described GAG composition with pharmaceutically acceptable carriers and/or excipients.
Claims Coverage
The partial claim set includes two independent composition claims focused on genetically engineered-cell-derived heparan sulfate compositions with defined purity constraints and altered interaction with FGF2, together with specified genetic modification scopes for producing cell lines.
Reduced FGF2 binding heparan sulfate composition with defined cell genetic modifications
A composition comprising a heparan sulfate having reduced FGF2 binding compared to a heparan sulfate derived from a wild-type CHO cell, wherein the heparan sulfate is derived from a cell line comprising genetic modifications selected from deficiency in HS2ST, deficiency in HSNDST1 and deficiency in HSNDST2, and transgenic for Sulfatase 2 (Sulf2), and genetic modifications selected from deficiency in ChSy, Chondroitin Sulfate N-Acetylgalactosaminyltransferase 2 (CSGalNAcT2), and Chondroitin Polymerizing Factor (ChPF), wherein the composition is at least 95% free from chondroitin sulfate.
Increased FGF2 cell surface binding or signaling heparan sulfate composition with defined cell genetic modifications
A composition comprising a heparan sulfate having increased FGF2 cell surface binding or FGF2 signaling compared to a heparan sulfate derived from a wild-type CHO cell, wherein the heparan sulfate is derived from a cell line comprising a genetic modification to overexpress a gene selected from HSNDST1, HSNDST2, HS6ST1, and HS6ST2, and be deficient in a gene selected from ChSy, CSGalNAcT2, and ChPF, wherein the composition is at least 95% free from chondroitin sulfate.
Across the independent claims, the inventive scope is defined by genetically engineered cell line modifications that alter heparan sulfate interaction with FGF2 while constraining the composition purity to be at least 95% free from chondroitin sulfate.
Stated Advantages
Provides heparan sulfate with reduced FGF2 binding compared to heparan sulfate derived from a wild-type CHO cell.
Provides heparan sulfate with increased FGF2 cell surface binding or FGF2 signaling compared to heparan sulfate derived from a wild-type CHO cell.
Meets purity constraints including being at least 95% free from chondroitin sulfate.
In some embodiments, is at least 95% free from protein and nucleic acid contamination and/or at least 95% free from specific contaminant GAGs.
Provides therapeutic effect outcomes including symptom improvement for microbial infection pathogens.
Provides glycosaminoglycan/heparan sulfate compositions with defined sulfation or defined modification patterns for disease treatment use-cases.
Documented Applications
Treating thrombosis.
Treating inflammation.
Treating cancer.
Treating microbial infection, including MRSA.
Treating neurodegenerative disorders.
Treating wound healing.
Treatment of genetic disorders using glycosaminoglycan/heparan sulfate compositions with defined modification patterns.
Treatment of wounds using glycosaminoglycan/heparan sulfate compositions with defined modification patterns.
Interested in licensing this patent?