The invention relates to a novel process for obtaining direct sulfation of unprotected sugars, in particular polysaccharides, using ecologically acceptable solvents.

The present invention includes methods for preparing anticoagulant polysaccharides using several non-naturally occurring, engineered sulfotransferase enzymes that are designed to react with aryl sulfate compounds instead of the natural substrate, PAPS, to facilitate sulfo group transfer to polysaccharide sulfo group acceptors. Suitable aryl sulfate compounds include, but are not limited to, p-nitrophenyl sulfate or 4-nitrocatechol sulfate. Anticoagulant polysaccharides produced by methods of the present invention comprise N-, 3-O-, 6-O-sulfated glucosamine residues and 2-O sulfated hexuronic acid residues, have comparable anticoagulant activity compared to commercially-available anticoagulant polysaccharides, and can be utilized to form truncated anticoagulant polysaccharides having a reduced molecular weight.

Systems, methods, and kits are described for repairing a fibrocartilage defect in a subject. The fibrocartilage defect is contacted with a first composition containing an oxidized and methacrylated glycosaminoglycan to form an imine bond between the glycosaminoglycan and the fibrocartilage defect, thereby coating the fibrocartilage defect with the glycosaminoglycan. The fibrocartilage defect coated with the glycosaminoglycan is then contacted with a mixture of a pre-polymer hydrogel composition containing a first crosslinking unit that, when polymerized, is capable of bonding to methacrylate and a hydrogel polymerization initiator composition, thereby forming a hydrogel that is covalently bonded to the glycosaminoglycan through methacrylate.

The present invention provides a 2-OST mutant exhibiting a high activity. Specifically, the present invention provides a 2-O-sulfation enzyme mutant, having a substitution of a leucine residue at position 321 with a basic amino acid residue in any one amino acid sequence of: (a) the amino acid sequence of SEQ ID NO: 2; (b) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence of SEQ ID NO: 2; (c) an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (d) the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (e) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (f) an amino acid sequence having 90% or more identity to the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; and having a 2-O-sulfate transfer activity.

The present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue and a method for producing a heparan sulfate with improved C5-epimerization efficiency. Specifically, the present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue, said method comprising producing the heparosan compound having the isomerized hexuronic acid residue from a heparosan compound in the presence of a protein selected from the group of consisting of the following (A) to (F): (A) a protein comprising the amino acid sequence of SEQ ID No:2; (B) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase activity; (D) a protein comprising the amino acid sequence of SEQ ID NO:5; (E) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity; and (F) a protein which comprises an amino acid sequence having one or several deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity.

A rapid testing mechanism for respiratory viral pathogens includes a filter material positioned to capture exhaled breath particles from a respiratory tract. A portion of the filter material is impregnated with a pathogen binding adsorptive reagent. When the exhaled breath particles pass through the filter material the following occurs: when the binding adsorptive reagent reacts, a positive test for respiratory viral pathogens is indicated by the filter material; and when pathogen binding adsorptive reagent does not react, a negative test for respiratory viral pathogens is indicated by the filter material.

The invention relates to a method for obtaining low molecular weight heparins which exhibit high stability. The method includes the treatment of depolymerized heparin with H.sub.2O.sub.2 in a ratio of between 0.04 and 1.0 liters of H.sub.2O.sub.2 (30-35% w/v in water) per kg of depolymerized heparin. The invention also relates to a low molecular weight heparin obtained by this method, pharmaceutical compositions of the same, and methods of treatment employing the same.

A skin-substitute is constructed by homogenizing an acellular dermal tissue matrix into a slurry, pouring the slurry into a mold, and lyophilizing the slurry.

The invention relates to a cosmetic process for caring for the skin, more particularly facial skin, in particular wrinkled skin, comprising the topical application to the skin of a cosmetic composition comprising a grafted polysaccharide polymer (I) and exposure of the treated skin to light radiation, polymer (I) being of formula:

PS—(CO—NH-L-X).sub.a(COOH)b

in which PS denotes the basic backbone of the polysaccharide bearing the carboxylic acid groups;
L is a divalent hydrocarbon-based group containing from 1 to 20 carbon atoms;
X denotes a photoactive group of azide or diazirine type;
a denotes the content of COOH groups substituted with the group —NH-L-X;
b denotes the content of unsubstituted free COOH groups;
a being between 0.01 and 0.8; b being between 0.2 and 0.99;
a+b=1

The invention also relates to the polymers (I) bearing a photoactive group X of diazirine type and to a composition comprising such a polymer in a physiologically acceptable medium.

The invention relates to agents that inhibit the binding of SARS-CoV-2 to SDCs or the endocytosis of SARS-CoV-2 with SDCs for use in the treatment of SARS-CoV-2 infection by inhibiting the cellular entry of SARS-CoV-2 and/or the SARS-CoV-2-induced inflammatory response. The invention relates further on to a method of inhibiting SARS-CoV-2 infection, the cellular entry of SARS-CoV-2, and the SARS-CoV-2-induced inflammatory response comprising contacting the SARS-CoV-2 target cells with an agent that inhibits the binding of SARS-CoV-2 to SDC or the endocytosis of SARS-CoV-2 with SDCs. The agent as SDC-specific mono- or polyclonal antibody used may be a. an antibody specific for the human SDC isoforms (SDC1, -2, -3, -4), or b. as SDC-specific peptides, peptide ligands that interact with the extracellular domain of human SDC isoforms (SDC1, -2, -3, -4), such as TAT (amino acid sequence YGRKKRRQRRR), penetratin (amino acid sequence RQIKIWFQNRRMKW), polyarginine (amino acid sequence RRRRRRRR) and their analogs of at least four amino acids in length, synthesized from D-, L- or other amino acid derivatives, or peptides and peptidomimetics interacting with the glycosaminoglycan side chains of SDCs, containing the conserved heparin-binding motif, PRRAR, or analogs of at least four amino acids in length, synthesized from D-, L- or other amino acid derivatives or conjugates of the former with other active agents and carriers (liposomes and other polymeric nanoparticles); or c. as SDC specific low- and high-molecular-weight ligands, any agents capable of binding to the extra- and intracellular domains of SDC, including the glycosaminoglycan side chains of SDCs; or d. as agents derived from SDCs, recombinant proteins derived from the extra- and intracellular domains of human SDC isoforms (SDC1, -2, -3, -4) and analogs synthesized from D-, L- or other amino acid derivatives of at least 4 amino acids in length, and derivatives of glycosaminoglycan side chains and glycosaminoglycan analogs of human SDC isoforms (SDC1, -2, -3, -4); or e. as agents reducing the expression of SDCs, nucleic acid-based agents capable of reducing the expression of SDCs and their conjugates and complexes packaged in viral and non-viral carriers (liposomes and other polymeric nanoparticles); or f. as agents containing the conserved intracellular domain of SDCs peptides, peptidomimetics and recombinant proteins of at least four amino acids in length, synthesized from D-, L- or other amino acid derivatives containing the sequence of any conserved region of the intracellular domain of human SDC isoforms (SDC1, -2, -3, -4); or g. as an inhibitor of endocytosis of syndecans, low molecular weight agents