Topically administered, skin-penetrating glycosaminoglycan formulations suitable for use in cosmetic and pharmaceutical applications

Abstract

The present invention relates to topical glycosaminoglycan compositions, particularly hyaluronan compositions, that facilitate the penetration of modified glycosaminoglycans through the skin barrier into the epidermal and dermal layers of the skin, thereby allowing for the dermal administration of a glycosaminoglycan, such as hyaluronan, without requiring an injection. Through their ability to deliver hyaluronan to the epidermal and dermal layers, the present formulations are therefore suitable for use in dermal rejuvenation, enhancement, hyaluronan replenishment and protection therapy. The glycosaminoglycan compositions are also useful as delivery devices to facilitate the dermal and transdermal delivery of cosmetically and pharmaceutically active substances, including pharmaceuticals, polypeptides, proteins and similarly sized biomacromolecules, through the skin barrier.

1 . A glycosaminoglycan composition comprising a glycosaminoglycan modified through the covalent linkage of a lipid moiety to about 1--15% of the repeating disaccharide monomer units of the giycosaminoglycan wherein:
 the glycosaminogiycan to be modified is hyaluronan, a hyaluronan derivative, a polysaccharide comprised of repeating disaccharide units of an uronic acid or hexose linked to a hexosamine, or derivatives thereof;   the glycosaminoglycan to be modified has a molecular weight in the range of about 2 kDa to about 2,500 kDa;   the lipid moiety comprises one or more naturally-occurring or synthetically-derived fatty acids, glycerolipids, phospholipids, sphingolipids, sterol lipids, pernol lipids, or derivatives thereof, provided that the lipid moiety contains a functional group on its polar head-group to allow for covalent linkage of the lipid to the glycosaminoglycan; and   the modified glycosaminoglycan is able to penetrate the skin harries or a mucous membrane when applied thereto,   
       whereby the modified glycosaminoglycan is formed by the reaction of the glycosaminoglycan with the lipid moiety. 
     
     
         2 . The glycosaminoglycan composition of  claim 1  wherein glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 1-12% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         3 . The glycosaminoglycan composition of  claim 1  wherein glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 1-10% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         4 . The glycosaminoglycan, composition of  claim 1  wherein giycosaminogiycan is modified through the covalent linkage of a lipid moiety to about 1-7.5% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         5 . The glycosaminoglycan composition of  claim 1  wherein glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 2-6% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         6 . The glycosaminoglycan compositions of any of  claims 1  to  5  wherein the glycosaminoglycan to be modified has a molecular weight in the range of about 50 kDa to about 2,500 kDa. 
     
     
         7 . The glycosaminoglycan composition of any of  claims 1  to  5  wherein the glycosaminoglycan has a molecule weight in the range of about 100 kDa to about 2,000 kDa. 
     
     
         8 . The glycosaminoglycan composition of any of  claims 1  to  5  wherein the glycosaminoglycan has a molecule weight in the range of about 350 kDa to about 1,500 kDa. 
     
     
         9 . The glycosaminoglycan composition of any of  claims 1  to  5  wherein the glycosaminoglycan has a molecule weight in the range of about 500 kDa to about 1,500 kDa. 
     
     
         10 . The glycosaminoglycan composition of any of  claim 1  or  9  wherein the glycosaminoglycan to be modified is hyaluronan or a hyaluronan derivative. 
     
     
         11 . The glycosaminoglycan composition of  claim 10  wherein the glycosaminoglycan to modified in hyaluronan. 
     
     
         12 . The glycosaminoglycan composition of any of  claims 1  to  11  wherein the lipid moiety contains en amino group on its polar head-group and is covalently bound to the glycosaminoglycan via an amide linkage to a carboxylic acid group on the glycosaminoglycan. 
     
     
         13 . The glycosaminoglycan composition of any of  claims 1  to  12  whereinthe lipid moiety comprises one or more phosphatidylethanolamines or phosphatidlyserines. 
     
     
         14 . The glycosaminoglycan composition of any of  claims 1  to  13  wherein the lipid moiety comprises one or more phosphatidylethanolamines. 
     
     
         15 . The glycosaminoglycan composition of any of  claims 1  to  14  wherein the modified glycosaminoglycan is able to penetrate the skit barrier. 
     
     
         16 . The glycosaminoglycan composition of any of  claims 1  to  15  wherein the modified glycosaminoglycan has a longer residence time within the skin than the unmodified glycosaminoglycan, when dermally delivered. 
     
     
         17 . The glycosaminoglycan composition of  claim 16  wherein the glycosaminoglycan to be modified is hyaluronan. 
     
     
         18 . A preparation suitable for application to the skin or a mucous membrane comprising a glycosaminoglycan composition in admixture with one or more cosmetically or pharmaceutically acceptable excipients or carriers, wherein the glycosaminoglycan composition comprises:
 a glycosaminoglycan modified through the covalent linkage of lipid moieties to 1-15% of the repeating disaccharide units monomer units of the glycosaminoglycan;   the glycosaminoglycan comprises hyaluronan, a hyaluronan derivative, a polysaccharide comprised of repeating disaccharide units of an uronic acid or hexose linked to a hexosamine, or derivatives thereof;   the glycosaminoglycan to be modified has a molecular weight in the range of about 2 kDa to about 2,500 kDa;   the lipid moiety comprises one or more, naturally-occurring or synthetically derived fatty acids, glycerolipids, phospholipids, sphingolipids, sterol lipids, prenol lipids, or derivatives thereof, provided that the lipid moiety contains a functional group on its polar head group to allow for covalent linkage of the lipid to the glycosaminoglycan; and   the modified glycosaminoglycan is able to penetrate the skin barrier or mucous membrane when applied thereto,   whereby the modified glycosaminoglycan formed by the reaction of the glycosaminoglycan with the lipid moiety.   
     
     
         19 . The preparation of  claim 18  wherein the glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 1-12% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         20 . The preparation of  claim 18  wherein the glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 1-10% of the repeating disaccharide monomer units of the glycosaminycan. 
     
     
         21 . The preparation of  claim 18  wherein the glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 1-7.5% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         22 . The preparation of  claim 18  wherein the glycosaminoglycan is modified through the covalent linkage of a lipid moiety to about 2-6% of the repeating disaccharide monomer units of the glycosaminoglycan. 
     
     
         23 . The preparation of any of  claims 18  to  22  wherein the glycosaminoglycan be modified has a molecular weight in the range of about 50 kDa to about 2,500 kDa. 
     
     
         24 . The preparation of any of  claims 18  to  22  wherein the glycosaminoglycan to be modified has a molecular weight in the range of about 100 kDa to about 2,000 kDa. 
     
     
         25 . The preparation of any of  claims 18  to  22  wherein the glycosaminoglycan to be modified has a molecular weight in the range of about 350 kDa to about 1,500 kDa. 
     
     
         26 . The preparation of any of  claims 18  to  22  wherein the glycosaminoglycan to be r modified has a molecular weight in the range of about 500 kDa to about 1,500 kDa. 
     
     
         27 . The preparation of any of  claims 18  to  26  herein the glyocsaminoglycan is hyaluronan or a hyaluronan derivative. 
     
     
         28 . The preparation of  claim 27  wherein the glycosaminoglycan is hyaluronan. 
     
     
         29 . The preparation of any of  claims 18  to  28  wherein the lipid moiety contains an amino group on its polar head-group and is covalently bound to the glycosaminoglycan via an amide linkage to a carboxylic acid group on the glycosaminoglycan. 
     
     
         30 . The preparation of any of  claims 18  to  29  wherein the lipid moiety comprises one or more phosphatidylethanolamines or phosphatidylserines. 
     
     
         31 . The preparation of any of  claims 18  to  30  wherein the lipid moiety comprises phosphatidylethanolamines. 
     
     
         32 . The preparation of any of  claims 18  to  31  wherein the preparation is suitable for the delivery of the modified glycosaminoglycan through the skin barrier. 
     
     
         33 . The preparation of any of  claims 18  to  32  herein the modified glycosaminoglycan has a longer residence time within the skin than the unmodified glycosaminoglycan, when dermally delivered. 
     
     
         34 . The preparation of any of  claims 18  to  33  additionally comprising one or more ingredients that are transported across the skin barrier or mucous membrane by the glycosaminoglycan composition. 
     
     
         35 . The preparation of  claim 34  wherein the additional one or more ingredients are transported across the skin barrier. 
     
     
         36 . The preparation of m  35  wherein the additional one or more ingredients comprises anti-oxidants, vitamins, essential oils, UV-blocking agents, or other nutrients whose application is known to provide a beneficial effect to the health or appearance of skin. 
     
     
         37 . The preparation of any of  claims 34  or  35  wherein the additional one or more ingredients is a pharmaceutical. 
     
     
         38 . The preparation of  claim 37  wherein the additional one or more ingredients comprises therapeutic agents suitable for the treatment inflammation, skin cancer or skin conditions that are delivered through the skin barrier. 
     
     
         39 . The preparation of  claim 38  wherein the additional one or more ingredients is suitable for the treatment of skin cancer. 
     
     
         40 . The preparation of  claim 38  wherein the additional one or more ingredients is suitable for the treatment of skin conditions. 
     
     
         41 . The preparation of  claim 37  wherein the additional one ingredients comprises an anti-inflammatory agent. 
     
     
         42 . The preparation of  claim 37  wherein the additional one or ingredients comprise a prostaglandin. 
     
     
         43 . The preparation of any of  claim 34  or  35  wherein the additional one or more ingredients comprises a protein, peptide, peptide mimetic, pepducin, polynucleotide, or other biomolecule. 
     
     
         44 . The preparation of  claim 43  wherein the additional one or more ingredients has a molecular weight of between about 700 Da and about 500 kDa. 
     
     
         45 . The preparation of any of  claim 43  or  44  wherein the additional one or more ingredients comprises collagen that is delivered across the skin barrier. 
     
     
         46 . The preparation of any of  claim 43  or  44  wherein the additional one or more ingredients comprises elastin that is delivered across the skin barrier. 
     
     
         47 . The preparation of  claim 44  wherein the additional one or more ingredients comprises a protein. 
     
     
         48 . The preparation of  claim 47  wherein the protein is a botulinum toxin that is delivered across the skin harrier. 
     
     
         49 . The preparation of  claim 48  wherein the protein is botulinum toxin type A. 
     
     
         50 . The preparation of  claim 47  wherein hyaluronan synthase is delivered across the skin barrier. 
     
     
         51 . The preparation of  claim 44  wherein the additional one or more ingredients comprises pepducin. 
     
     
         52 . The preparation of  claim 34  wherein the additional one or more ingredients comprises a hyaluronidase inhibitor that is delivered across the skin barrier. 
     
     
         53 . The preparation of  claim 34  wherein the additional or more ingredients comprises a RHAMM inhibitor that is delivered across the skin barrier. 
     
     
         54 - 88 . (canceled) 
     
     
         89 . A method for the manufacture of a glycosaminoglycan composition as described in any of  claims 1  to  17  comprising the steps of:
 treating the glycosaminoglycan to be modified with an activating agent to facilitate covalent bonding of the glycosaminoglycan to the lipid moiety; 
 mixing the activated glycosaminoglycan and lipid moiety; and 
 allowing the lipid moiety to react with the activated glycosaminoglycan to covalently link the lipid moiety to the glycosaminoglycan, 
 wherein the activating agent is the limiting reagent in the reaction and is added in an amount sufficient to facilitate covalent linkage of the lipid moiety to about 1 to about 15% of the disaccharide monomer units of the glycosaminoglycan. 
 
     
     
         90 . The method according to  claim 89  wherein the activating agent is added in an amount sufficient to facilitate covalent linkage of the lipid moiety to about 1 to about 12% of the disaccharide monomer units of the glycosaminoglycan. 
     
     
         91 . The method according to  claim 89  wherein the activating agent is added in an amount sufficient to facilitate covalent linkage of the lipid moiety to about 1 to about 10% of the disaccharide monomer units of the glycosaminoglycan. 
     
     
         92 . The method according to  claim 89  wherein the activating agent is added in an amount sufficient to facilitate covalent linkage of the lipid moiety to about 1 to about 7.5% of the disaccharide monomer units of the glycosaminoglycan. 
     
     
         93 . The method according to  claim 89  wherein the activating agent is added in an amount sufficient to facilitate covalent linkage of the lipid moiety to about 2 to about 6% of the disaccharide monomer units of the glycosaminoglycan. 
     
     
         94 . The method according to any of  claims 89  to  93  wherein the covalent linkage to be formed is an amide linkage between a carboxylic group of the glycosaminoglycan and a lipid moiety containing an amine functional group on the polar head group. 
     
     
         95 . The method according to any of  claims 89  end  94  wherein the activating agent is a carbodiimide. 
     
     
         96 . The method according to  claim 95  wherein the carbodiimide is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. 
     
     
         97 . The method according to any of  claims 89  to  96  wherein the glycosaminoglycan to be modified is hyaluronan. 
     
     
         98 . The method according to any of  claims 89  to  97  wherein the lipid moiety comprises one or more phosphatidylethanolamines and phosphatidylserines. 
     
     
         99 . The method according to any of  claims 89  to  97  wherein the lipid moiety comprises one or more phosphatidylethanolamines. 
     
     
         100 . The glycosaminoglycan composition of  claim 1  wherein the modified glycosaminoglycan is a non-particulate hyaluronan-phospholipid. 
     
     
         101 . The glycosaminoglycan composition of  claim 100  wherein the non-particulate hyaluronan-phospholipid has an increased residence time within the skin as an intact hyaluronan-phospholipid. 
     
     
         102 - 104 . (canceled)