Rapid In-situ Gel Forming Compositions Prepared via Reaction of Vinyl Sulfone Derivatized Hyaluronic Acid with Thiol Derivatized Polyethylene Glycol at Alkaline pH

Abstract

The present application provides rapid-gelling, sprayable hyaluronic-acid based compositions, kits, related methods, precursor formulations, and uses thereof.

1 . A kit comprising:
 (i) a first container comprising an aqueous solution of vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) at a concentration of from about 10-300 mg/mL, where HA-VS is (2-(vinylsulfonyl)ethoxy)hyaluronic acid, and the HA-VS has from about 2%-70% of its hydroxyl groups transformed to 2-(vinylsulfonyl)ethoxy groups,   (ii) a second container comprising a thiol-functionalized polyethylene glycol having from 2 to 8 thiol groups, and   (iii) a third container comprising a 30-1000 mM buffer solution at a pH ranging from about 7-12, in an amount effective, when mixed with the contents of the first and second containers, to provide a solution having a HA-VS concentration of 2-8% (w/v) and a thiol-functionalized polyethylene glycol concentration of 2-8% (w/v),   wherein the components of the first, second and third containers, when combined, are effective to form a gel within from about 5 seconds to 1 hour of mixing.   
     
     
         2 . A liquid composition formed from the combination of (i) an aqueous solution of vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) at a concentration of from about 10-300 mg/mL, where where HA-VS is (2-(vinylsulfonyl)ethoxy)hyaluronic acid, and the HA-VS has from about 2%-70% of its hydroxyl groups transformed to 2-(vinylsulfonyl)ethoxy groups, (ii) a thiol-functionalized polyethylene glycol having from 2 to 8 thiol groups, and (iii) a 30-1000 mM buffer solution at a pH ranging from about 7-12,
 where the concentration of the thiol-functionalized polyethylene glycol in the liquid composition ranges from about 4-300 mg/mL, and 
 the liquid composition is effective to form a gel within from about 5 seconds to about 1 hour of mixing components (i), (ii) and (iii). 
 
     
     
         3 . The kit of  claim 1 , wherein the vinyl sulfone-derivatized hyaluronic acid has from about 7%-35%, of its hydroxyl groups substituted with vinyl sulfone. 
     
     
         4 . The kit of  claim 1 , wherein the thiol-functionalized polyethylene glycol has a number of thiol groups selected from the group consisting of 2, 3, and 4. 
     
     
         5 . The kit of  claim 1 , wherein the vinyl sulfone-derivatized hyaluronic acid has an average molecular weight ranging from about 15,000 to about 1,000,000 daltons. 
     
     
         6 . The kit of  claim 1 , wherein the thiol-functionalized polyethylene glycol is linear or branched. 
     
     
         7 . The kit of  claim 1 , wherein the thiol-functionalized polyethylene glycol has an average molecular weight of from about 1,000 to about 10,000 daltons. 
     
     
         8 . The kit of  claim 1 , wherein the molecular weight of the thiol-functionalized polyethylene glycol is less than the molecular weight of the vinyl sulfone-derivatized hyaluronic acid. 
     
     
         9 . The kit of  claim 1 , wherein the aqueous solution of vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) has a concentration ranging from about 20-200 mg/mL. 
     
     
         10 . The kit of  claim 1 , wherein the thiol-functionalized polyethylene glycol is in the form of a powder. 
     
     
         11 . The kit of  claim 1 , wherein the amount of thiol-functionalized polyethylene glycol relative to vinyl sulfone-derivatized hyaluronic acid ranges from about 1:1 (w/w) to about 0.4:1 (w/w). 
     
     
         12 . The kit of  claim 1 , wherein at least one of:
 one of the first, second or third containers further comprises a bioactive agent; and   the kit further comprises a fourth container comprising a bioactive agent.   
     
     
         13 . The liquid composition of  claim 2 , further comprising a bioactive agent. 
     
     
         14 . The liquid composition of  claim 13 , wherein the bioactive agent is a corticosteroid selected from the group consisting of triamcinolone, triamcinolone acetonide or triamcinolone hexacetonide. 
     
     
         15 . The kit of  claim 12 , wherein the bioactive agent is a corticosteroid selected from the group consisting of triamcinolone, triamcinolone acetonide and triamcinolone hexacetonide. 
     
     
         16 . The kit of  claim 1 , wherein the first container further comprises from about 0.1 weight % to about 3.5 weight percent of the thiol-functionalized polyethylene glycol relative to the vinyl sulfone-derivatized hyaluronic acid (w/w). 
     
     
         17 . A method of forming a liquid composition capable of in-situ gel formation, comprising:
 (i) adding a thiol-functionalized polyethylene glycol having from 2 to 8 thiol groups to a aqueous solution of vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) having a concentration of from about 4-300 mg/mL, where HA-VS is (2-(vinylsulfonyl)ethoxy)hyaluronic acid, and the HA-VS has from about 2% to 70% of its hydroxyl groups transformed to 2-(vinylsulfonyl)ethoxy groups, to thereby dissolve the thiol-functionalized polyethylene glycol to form a vinyl sulfone-derivatized hyaluronic acid-thiol-functionalized polyethylene glycol-containing solution, and   (ii) combining the solution from step (i) with a 30-1000 mM buffer solution at a pH ranging from about 7-12, to thereby form a liquid composition having a HA-VS concentration of 2-8% (w/v) and a thiol-functionalized polyethylene glycol concentration of 2-8% (w/v),   wherein the liquid composition is effective to form a gel within from about 5 seconds to about 1 hour of combining.   
     
     
         18 . The method of  claim 17 , wherein:
 (i) the vinyl sulfone-derivatized hyaluronic acid has from about 7% to 35% of its hydroxyl groups substituted with vinyl sulfone; and/or   (ii) the vinyl sulfone-derivatized hyaluronic acid has an average molecular weight selected from a range of from about 10,000 to about 2,000,000 daltons; a range of from about 15,000 to about 1,000,000 daltons, a range from about 20,000 to about 200,000 daltons; or has an average molecular weight of about 100,000 daltons, and/or   (iii) the aqueous solution of the vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) has a concentration ranging from about 20-200 mg/mL; and/or   (iv) the molecular weight of the thiol-functionalized polyethylene glycol is less than the molecular weight of the vinyl sulfone-derivatized hyaluronic acid; and/or   (v) the amount of thiol-functionalized polyethylene glycol relative to vinyl sulfone-derivatized hyaluronic acid ranges from about 1:1 (w/w) to about 0.4:1 (w/w); and/or   (vi) the buffer solution has a pH ranging from about 8.0 to 10.5.   
     
     
         19 . The method of  claim 17  further comprising adding a bioactive agent to the vinyl sulfone-derivatized hyaluronic acid-thiol-functionalized polyethylene glycol-containing solution of step (i), or to the buffer solution from step (ii) prior to the combining, or to the liquid composition formed in step (ii). 
     
     
         20 . The method of  claim 19 , wherein the bioactive agent is a corticosteroid selected from the group consisting of triamcinolone, triamcinolone acetonide or triamcinolone hexacetonide. 
     
     
         21 . A method of forming a liquid composition capable of in-situ gel formation, comprising:
 (i) adding a portion of an overall amount of thiol-functionalized polyethylene glycol to an aqueous solution of vinyl sulfone-derivatized hyaluronic acid (“HA-VS”) having a concentration of from about 10-300 mg/mL, where HA-VS is (2-(vinylsulfonyl)ethoxy)hyaluronic acid, and the HA-VS has from about 2%-70% of its hydroxyl groups transformed to 2-(vinylsulfonyl)ethoxy groups, to thereby form a solution,   (ii) sterilizing the solution from step (i),   (iii) adding to the solution from step (i) or step (ii), the remaining amount of thiol-functionalized polyethylene glycol, where the thiol-functionalized polyethylene glycol has from 2 to 8 thiol groups, to thereby dissolve the remaining amount of thiol-functionalized polyethylene glycol to form a vinyl sulfone-derivatized hyaluronic acid-thiol-functionalized polyethylene glycol-containing solution, and   (iv) mixing the vinyl sulfone-derivatized hyaluronic acid-thiol-functionalized polyethylene glycol-containing solution with a 30 -1000 mM buffer solution at a pH ranging from about 7-12,   to thereby form a liquid composition having a HA-VS concentration of 2-8% (w/v) and a thiol-functionalized polyethylene glycol concentration of 2-8% (w/v), to thereby form a liquid composition effective to form a gel within from about 5 seconds to about 1 hour of mixing.   
     
     
         22 . The method of  claim 21 , wherein the portion of thiol-functionalized polyethylene glycol in step (i) comprises from about 0.1 weight % to about 3.5 weight percent thiol-functionalized polyethylene glycol relative to the vinyl sulfone-derivatized hyaluronic acid (w/w). 
     
     
         23 . The method of  claim 21 , further comprising adding a bioactive agent to the solution of step (i), or to the vinyl sulfone-derivatized hyaluronic acid-thiol-functionalized polyethylene glycol-containing solution from step (iii), or to the buffer solution from step (iv) prior to mixing, or to the liquid composition formed in step (iv). 
     
     
         24 . The method of  claim 23 , wherein the bioactive agent is a corticosteroid selected from the group consisting of triamcinolone, triamcinolone acetonide and triamcinolone hexacetonide. 
     
     
         25 . A method for treating a condition selected from the group consisting of osteoarthritis, rheumatoid arthritis, a wound, and prevention of surgical adhesions, comprising applying the liquid composition of  claim 2  to a body site in a subject suffering from the condition. 
     
     
         26 . The kit of  claim 1 , wherein one or more of the kit components is sterile. 
     
     
         27 . The liquid composition of  claim 2 , having one or more components in sterile form. 
     
     
         28 . The method of  claim 21 , comprising (ii) sterilizing the solution from step (i) and/or where either one or both of the thiol-functionalized polyethylene glycol from step (iii) and the buffer solution is sterile. 
     
     
         29 . A hydrogel formed from the liquid composition of  claim 2 . 
     
     
         30 . A method for repairing tissue of a joint during arthroscopic or open joint surgery by applying the liquid composition of  claim 2  to the tissue.