US20110319357A1 - Hyaluronic Acid Derivatives - Google Patents
Hyaluronic Acid Derivatives Download PDFInfo
- Publication number
- US20110319357A1 US20110319357A1 US13/164,012 US201113164012A US2011319357A1 US 20110319357 A1 US20110319357 A1 US 20110319357A1 US 201113164012 A US201113164012 A US 201113164012A US 2011319357 A1 US2011319357 A1 US 2011319357A1
- Authority
- US
- United States
- Prior art keywords
- hyaluronic acid
- acid derivative
- osa
- alkyl
- asa
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical class CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 title claims abstract description 192
- -1 aryl/alkyl succinic anhydrides Chemical class 0.000 claims abstract description 40
- 239000000203 mixture Substances 0.000 claims abstract description 23
- 239000002537 cosmetic Substances 0.000 claims abstract description 19
- 239000003814 drug Substances 0.000 claims abstract description 13
- 239000003094 microcapsule Substances 0.000 claims abstract description 5
- 239000004005 microsphere Substances 0.000 claims abstract description 3
- 239000002077 nanosphere Substances 0.000 claims abstract description 3
- 125000000217 alkyl group Chemical group 0.000 claims description 26
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 8
- 239000001384 succinic acid Substances 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 5
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N lactose group Chemical group OC1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@@H](O)[C@H](O2)CO)[C@H](O1)CO GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 5
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 5
- 206010052428 Wound Diseases 0.000 claims description 4
- 208000027418 Wounds and injury Diseases 0.000 claims description 4
- 239000008101 lactose Substances 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 125000003837 (C1-C20) alkyl group Chemical group 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 3
- 239000002088 nanocapsule Substances 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 230000029663 wound healing Effects 0.000 claims description 3
- 125000004185 ester group Chemical group 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 229920002674 hyaluronan Polymers 0.000 abstract description 156
- 229960003160 hyaluronic acid Drugs 0.000 abstract description 137
- 230000004048 modification Effects 0.000 abstract description 6
- 238000012986 modification Methods 0.000 abstract description 6
- 230000015572 biosynthetic process Effects 0.000 abstract description 5
- 229940014800 succinic anhydride Drugs 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 4
- 238000009472 formulation Methods 0.000 abstract description 4
- 238000005538 encapsulation Methods 0.000 abstract description 2
- 230000037368 penetrate the skin Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 43
- 239000000243 solution Substances 0.000 description 42
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 39
- 239000000047 product Substances 0.000 description 32
- 238000000034 method Methods 0.000 description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 23
- 238000013019 agitation Methods 0.000 description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 20
- WVRNUXJQQFPNMN-VAWYXSNFSA-N 3-[(e)-dodec-1-enyl]oxolane-2,5-dione Chemical compound CCCCCCCCCC\C=C\C1CC(=O)OC1=O WVRNUXJQQFPNMN-VAWYXSNFSA-N 0.000 description 19
- 241000193830 Bacillus <bacterium> Species 0.000 description 19
- KIUKXJAPPMFGSW-MNSSHETKSA-N hyaluronan Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H](C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-MNSSHETKSA-N 0.000 description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 18
- 229940099552 hyaluronan Drugs 0.000 description 18
- 150000003839 salts Chemical class 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 14
- HDFKMLFDDYWABF-UHFFFAOYSA-N 3-phenyloxolane-2,5-dione Chemical compound O=C1OC(=O)CC1C1=CC=CC=C1 HDFKMLFDDYWABF-UHFFFAOYSA-N 0.000 description 13
- 239000000523 sample Substances 0.000 description 12
- 239000012901 Milli-Q water Substances 0.000 description 11
- 125000003118 aryl group Chemical group 0.000 description 11
- FGQUIQAGZLBOGL-UHFFFAOYSA-N 3-non-1-enyloxolane-2,5-dione Chemical compound CCCCCCCC=CC1CC(=O)OC1=O FGQUIQAGZLBOGL-UHFFFAOYSA-N 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 10
- 238000006467 substitution reaction Methods 0.000 description 10
- KLAIOABSDQUNSA-WUKNDPDISA-N 3-[(e)-octadec-2-enyl]oxolane-2,5-dione Chemical compound CCCCCCCCCCCCCCC\C=C\CC1CC(=O)OC1=O KLAIOABSDQUNSA-WUKNDPDISA-N 0.000 description 9
- 229920002683 Glycosaminoglycan Polymers 0.000 description 9
- 0 [1*]C1C([2*])C(OC2C([3*])C(C[4*])OC(C)C2NC(C)=O)OC(C(=O)[O-])C1OC Chemical compound [1*]C1C([2*])C(OC2C([3*])C(C[4*])OC(C)C2NC(C)=O)OC(C(=O)[O-])C1OC 0.000 description 9
- 238000000502 dialysis Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 239000011780 sodium chloride Substances 0.000 description 9
- 239000004480 active ingredient Substances 0.000 description 8
- 230000002776 aggregation Effects 0.000 description 8
- 238000004220 aggregation Methods 0.000 description 8
- 150000002148 esters Chemical class 0.000 description 8
- 238000004108 freeze drying Methods 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 239000000693 micelle Substances 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 244000063299 Bacillus subtilis Species 0.000 description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- WCDDVEOXEIYWFB-VXORFPGASA-N (2s,3s,4r,5r,6r)-3-[(2s,3r,5s,6r)-3-acetamido-5-hydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5,6-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@@H]1C[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O)[C@H](O)[C@H]1O WCDDVEOXEIYWFB-VXORFPGASA-N 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 6
- 102000003918 Hyaluronan Synthases Human genes 0.000 description 6
- 108090000320 Hyaluronan Synthases Proteins 0.000 description 6
- 229920000881 Modified starch Polymers 0.000 description 6
- 239000013543 active substance Substances 0.000 description 6
- 239000000839 emulsion Substances 0.000 description 6
- 229940014041 hyaluronate Drugs 0.000 description 6
- 235000019426 modified starch Nutrition 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 229920002472 Starch Polymers 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 235000019698 starch Nutrition 0.000 description 5
- 201000004384 Alopecia Diseases 0.000 description 4
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 4
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 4
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 4
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 102000016611 Proteoglycans Human genes 0.000 description 4
- 108010067787 Proteoglycans Proteins 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- IAJILQKETJEXLJ-QTBDOELSSA-N aldehydo-D-glucuronic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-QTBDOELSSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 4
- 239000004327 boric acid Substances 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000012869 ethanol precipitation Methods 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 239000006260 foam Substances 0.000 description 4
- 229940097043 glucuronic acid Drugs 0.000 description 4
- 230000003676 hair loss Effects 0.000 description 4
- 239000000017 hydrogel Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 210000003491 skin Anatomy 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 238000000108 ultra-filtration Methods 0.000 description 4
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 3
- 241001328122 Bacillus clausii Species 0.000 description 3
- 241000193422 Bacillus lentus Species 0.000 description 3
- 241000194108 Bacillus licheniformis Species 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- OQILCOQZDHPEAZ-UHFFFAOYSA-N Palmitinsaeure-octylester Natural products CCCCCCCCCCCCCCCC(=O)OCCCCCCCC OQILCOQZDHPEAZ-UHFFFAOYSA-N 0.000 description 3
- 150000008064 anhydrides Chemical class 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 238000001212 derivatisation Methods 0.000 description 3
- 150000002016 disaccharides Chemical group 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- GJQLBGWSDGMZKM-UHFFFAOYSA-N ethylhexyl palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC(CC)CCCCC GJQLBGWSDGMZKM-UHFFFAOYSA-N 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000004340 gradient COSY Methods 0.000 description 3
- 239000011859 microparticle Substances 0.000 description 3
- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 3
- NFBVOAPHAOKEGN-UHFFFAOYSA-N 3,3,4,4-tetrapropyloxolane-2,5-dione Chemical compound CCCC1(CCC)C(=O)OC(=O)C1(CCC)CCC NFBVOAPHAOKEGN-UHFFFAOYSA-N 0.000 description 2
- RSPWVGZWUBNLQU-FOCLMDBBSA-N 3-[(e)-hexadec-1-enyl]oxolane-2,5-dione Chemical compound CCCCCCCCCCCCCC\C=C\C1CC(=O)OC1=O RSPWVGZWUBNLQU-FOCLMDBBSA-N 0.000 description 2
- KAYAKFYASWYOEB-UHFFFAOYSA-N 3-octadec-1-enyloxolane-2,5-dione Chemical compound CCCCCCCCCCCCCCCCC=CC1CC(=O)OC1=O KAYAKFYASWYOEB-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 description 2
- 229920002261 Corn starch Polymers 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 229920000288 Keratan sulfate Polymers 0.000 description 2
- OVRNDRQMDRJTHS-RTRLPJTCSA-N N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-RTRLPJTCSA-N 0.000 description 2
- 229920002385 Sodium hyaluronate Polymers 0.000 description 2
- 241000264435 Streptococcus dysgalactiae subsp. equisimilis Species 0.000 description 2
- 241000120569 Streptococcus equi subsp. zooepidemicus Species 0.000 description 2
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 2
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000033115 angiogenesis Effects 0.000 description 2
- 150000001642 boronic acid derivatives Chemical class 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 238000007707 calorimetry Methods 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 210000001520 comb Anatomy 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000002500 effect on skin Effects 0.000 description 2
- 238000004945 emulsification Methods 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 208000024963 hair loss Diseases 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- KXCLCNHUUKTANI-RBIYJLQWSA-N keratan Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@H](COS(O)(=O)=O)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@H](O[C@@H](O[C@H]3[C@H]([C@@H](COS(O)(=O)=O)O[C@@H](O)[C@@H]3O)O)[C@H](NC(C)=O)[C@H]2O)COS(O)(=O)=O)O[C@H](COS(O)(=O)=O)[C@@H]1O KXCLCNHUUKTANI-RBIYJLQWSA-N 0.000 description 2
- 229950006780 n-acetylglucosamine Drugs 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 125000004365 octenyl group Chemical group C(=CCCCCCC)* 0.000 description 2
- 201000008482 osteoarthritis Diseases 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000009877 rendering Methods 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 2
- 229940010747 sodium hyaluronate Drugs 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- AYCBRUDZNRVKGK-GWLSAQFNSA-N (2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(4R,10S,16S,19S,22S,28S,31S,34S,37S,40S,49S,52R)-52-[[(2S)-2-[[(2S)-2-amino-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-19-(3-amino-3-oxopropyl)-49-benzyl-28,37-bis[(2S)-butan-2-yl]-31,40-bis(3-carbamimidamidopropyl)-34-(carboxymethyl)-16-(hydroxymethyl)-22-methyl-10-(2-methylpropyl)-6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51-hexadecaoxo-1,2-dithia-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50-hexadecazacyclotripentacontane-4-carbonyl]amino]-4-oxobutanoyl]amino]-3-hydroxypropanoyl]amino]-3-phenylpropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)=O)[C@@H](C)CC)[C@@H](C)CC)C1=CC=CC=C1 AYCBRUDZNRVKGK-GWLSAQFNSA-N 0.000 description 1
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- LVFFZQQWIZURIO-UHFFFAOYSA-N 2-phenylbutanedioic acid Chemical compound OC(=O)CC(C(O)=O)C1=CC=CC=C1 LVFFZQQWIZURIO-UHFFFAOYSA-N 0.000 description 1
- FLISWPFVWWWNNP-UHFFFAOYSA-N 3-oct-1-enyloxolane-2,5-dione Chemical group CCCCCCC=CC1CC(=O)OC1=O FLISWPFVWWWNNP-UHFFFAOYSA-N 0.000 description 1
- WSGFXVFLWVXTCJ-UHFFFAOYSA-N 3-oct-2-enyloxolane-2,5-dione Chemical compound CCCCCC=CCC1CC(=O)OC1=O WSGFXVFLWVXTCJ-UHFFFAOYSA-N 0.000 description 1
- GUOCOOQWZHQBJI-UHFFFAOYSA-N 4-oct-7-enoxy-4-oxobutanoic acid Chemical group OC(=O)CCC(=O)OCCCCCCC=C GUOCOOQWZHQBJI-UHFFFAOYSA-N 0.000 description 1
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 102100033367 Appetite-regulating hormone Human genes 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 208000036487 Arthropathies Diseases 0.000 description 1
- 244000186140 Asperula odorata Species 0.000 description 1
- 102400001276 Atriopeptin-3 Human genes 0.000 description 1
- 241000193752 Bacillus circulans Species 0.000 description 1
- 241000193749 Bacillus coagulans Species 0.000 description 1
- 241000193747 Bacillus firmus Species 0.000 description 1
- 241000194107 Bacillus megaterium Species 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 241000193764 Brevibacillus brevis Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- 102100023804 Coagulation factor VII Human genes 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 229920000045 Dermatan sulfate Polymers 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 108090000394 Erythropoietin Proteins 0.000 description 1
- 102000003951 Erythropoietin Human genes 0.000 description 1
- 108010023321 Factor VII Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 235000008526 Galium odoratum Nutrition 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 1
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 102000038461 Growth Hormone-Releasing Hormone Human genes 0.000 description 1
- 239000000095 Growth Hormone-Releasing Hormone Substances 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 108010000521 Human Growth Hormone Proteins 0.000 description 1
- 102000002265 Human Growth Hormone Human genes 0.000 description 1
- 239000000854 Human Growth Hormone Substances 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- AEMOLEFTQBMNLQ-HNFCZKTMSA-N L-idopyranuronic acid Chemical compound OC1O[C@@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-HNFCZKTMSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 235000019759 Maize starch Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 1
- 229910019093 NaOCl Inorganic materials 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000194109 Paenibacillus lautus Species 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 101710167959 Putative UTP-glucose-1-phosphate uridylyltransferase Proteins 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 101710142969 Somatoliberin Proteins 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 241000193996 Streptococcus pyogenes Species 0.000 description 1
- 101100394256 Streptococcus pyogenes hasC gene Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 description 1
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102100038834 UTP-glucose-1-phosphate uridylyltransferase Human genes 0.000 description 1
- COQLPRJCUIATTQ-UHFFFAOYSA-N Uranyl acetate Chemical compound O.O.O=[U]=O.CC(O)=O.CC(O)=O COQLPRJCUIATTQ-UHFFFAOYSA-N 0.000 description 1
- 102000057144 Uridine Diphosphate Glucose Dehydrogenase Human genes 0.000 description 1
- 108010054269 Uridine Diphosphate Glucose Dehydrogenase Proteins 0.000 description 1
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 1
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 229920001284 acidic polysaccharide Polymers 0.000 description 1
- 150000004805 acidic polysaccharides Chemical class 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000012382 advanced drug delivery Methods 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 230000019552 anatomical structure morphogenesis Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000001153 anti-wrinkle effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009697 arginine Nutrition 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 238000000429 assembly Methods 0.000 description 1
- 230000000712 assembly Effects 0.000 description 1
- 108010034646 atrial natriuretic factor prohormone (103-126) Proteins 0.000 description 1
- 229940090047 auto-injector Drugs 0.000 description 1
- 229940054340 bacillus coagulans Drugs 0.000 description 1
- 229940005348 bacillus firmus Drugs 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- PXTQQOLKZBLYDY-UHFFFAOYSA-N bis(2-ethylhexyl) carbonate Chemical compound CCCCC(CC)COC(=O)OCC(CC)CCCC PXTQQOLKZBLYDY-UHFFFAOYSA-N 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 108010006025 bovine growth hormone Proteins 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 125000000609 carbazolyl group Chemical class C1(=CC=CC=2C3=CC=CC=C3NC12)* 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229940059329 chondroitin sulfate Drugs 0.000 description 1
- 238000011098 chromatofocusing Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 238000002316 cosmetic surgery Methods 0.000 description 1
- AVJBPWGFOQAPRH-FWMKGIEWSA-L dermatan sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS([O-])(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C([O-])=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-L 0.000 description 1
- 229940051593 dermatan sulfate Drugs 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- FLISWPFVWWWNNP-BQYQJAHWSA-N dihydro-3-(1-octenyl)-2,5-furandione Chemical compound CCCCCC\C=C\C1CC(=O)OC1=O FLISWPFVWWWNNP-BQYQJAHWSA-N 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 229940105423 erythropoietin Drugs 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 229940012413 factor vii Drugs 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 108010077689 gamma-aminobutyryl-2-methyltryptophyl-2-methyltryptophyl-2-methyltryptophyl-lysinamide Proteins 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 101150097869 hasA gene Proteins 0.000 description 1
- 101150039161 hasB gene Proteins 0.000 description 1
- 101150068911 hasC1 gene Proteins 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 210000003035 hyaline cartilage Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- VJVOFLWZDWLHNR-MRCUWXFGSA-N icosan-9-yl (z)-docos-13-enoate Chemical compound CCCCCCCCCCCC(CCCCCCCC)OC(=O)CCCCCCCCCCC\C=C/CCCCCCCC VJVOFLWZDWLHNR-MRCUWXFGSA-N 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012678 infectious agent Substances 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 230000001050 lubricating effect Effects 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 235000018977 lysine Nutrition 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000013310 margarine Nutrition 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 235000010746 mayonnaise Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 238000013048 microbiological method Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000000921 morphogenic effect Effects 0.000 description 1
- 238000000569 multi-angle light scattering Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- HPZHFGBKCGWNGN-UHFFFAOYSA-N n-benzyl-2-methyl-7h-pyrrolo[2,3-d]pyrimidin-4-amine Chemical compound C=12C=CNC2=NC(C)=NC=1NCC1=CC=CC=C1 HPZHFGBKCGWNGN-UHFFFAOYSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000007764 o/w emulsion Substances 0.000 description 1
- 150000002482 oligosaccharides Polymers 0.000 description 1
- 238000010915 one-step procedure Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 150000002895 organic esters Chemical class 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000012667 polymer degradation Methods 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000004513 sizing Methods 0.000 description 1
- 230000008591 skin barrier function Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 238000010563 solid-state fermentation Methods 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 150000003900 succinic acid esters Chemical class 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 229960000187 tissue plasminogen activator Drugs 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 102000003390 tumor necrosis factor Human genes 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000003954 umbilical cord Anatomy 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 229960005356 urokinase Drugs 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/11—Encapsulated compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q7/00—Preparations for affecting hair growth
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/41—Particular ingredients further characterized by their size
- A61K2800/412—Microsized, i.e. having sizes between 0.1 and 100 microns
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/41—Particular ingredients further characterized by their size
- A61K2800/413—Nanosized, i.e. having sizes below 100 nm
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Rheumatology (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Cardiology (AREA)
- Ophthalmology & Optometry (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Heart & Thoracic Surgery (AREA)
- Physical Education & Sports Medicine (AREA)
- Pain & Pain Management (AREA)
- Toxicology (AREA)
- Vascular Medicine (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Cosmetics (AREA)
Abstract
The present invention relates to the modification of hyaluronic acid (HA) with aryl/alkyl succinic anhydrides (ASA) to produce aryl/alkyl succinic anhydride HA derivatives, to the derivatives as such, and to their applications and uses, particularly in the cosmetic and biomedical industries. The ASA-HA derivatives are expected to have interesting properties that can be used for advanced formulation (bind stronger to the skin compared to non-modified HA), possibly also in delivery systems for actives or drugs by encapsulation (nano/micro capsules) or formation of nano/micro spheres. Further, the low MW ASA-HA derivatives are expected to penetrate the skin more efficiently than non-modified HA of the same MW.
Description
- This application is a division of U.S. application Ser. No. 11/572,954 filed Jan. 30, 2007, now allowed, which is a 35 U.S.C. 371 national application of PCT/US2006/000523 filed Sep. 26, 2006, which claims priority or the benefit under 35 U.S.C. 119 of Danish application no. PA 2005 01332 filed Sep. 26, 2005 and U.S. provisional application No. 60/721,232 filed Sep. 27, 2005, the contents of which are fully incorporated herein by reference.
- The present invention relates to the modification of hyaluronic acid (HA) with aryl- or alkyl succinic anhydride (ASA) to produce aryl/alkyl succinic anhydride HA derivatives (ASA-HA), to the ASA-HA derivatives as such, and to their applications and uses, particularly in the cosmetics and biomedical industries.
- The most abundant heteropolysaccharides of the body are the glycosaminoglycans. Glycosaminoglycans are unbranched carbohydrate polymers, consisting of repeating disaccharide units (only keratan sulphate is branched in the core region of the carbohydrate). The disaccharide units generally comprise, as a first saccharide unit, one of two modified sugars —N-acetylgalactosamine (GalNAc) or N-acetylglucosamine (GlcNAc). The second unit is usually an uronic acid, such as glucuronic acid (GlcUA) or iduronate.
- Glycosaminoglycans are negatively charged molecules, and have an extended conformation that imparts high viscosity when in solution. Glycosaminoglycans are located primarily on the surface of cells or in the extracellular matrix. Glycosaminoglycans also have low compressibility in solution and, as a result, are ideal as a physiological lubricating fluid, e.g., joints. The rigidity of glycosaminoglycans provides structural integrity to cells and provides passageways between cells, allowing for cell migration. The glycosaminoglycans of highest physiological importance are hyaluronan, chondroitin sulfate, heparin, heparan sulfate, dermatan sulfate, and keratan sulfate. Most glycosaminoglycans bind covalently to a proteoglycan core protein through specific oligosaccharide structures. Hyaluronan forms large aggregates with certain proteoglycans, but is an exception as free carbohydrate chains form non-covalent complexes with proteoglycans.
- Numerous roles of hyaluronan in the body have been identified (see, Laurent and Fraser, 1992, FASEB J. 6: 2397-2404; and Toole B. P., 1991, “Proteoglycans and hyaluronan in morphogenesis and differentiation.” In: Cell Biology of the Extracellular Matrix, pp. 305-341, Hay E. D., ed., Plenum, N.Y.). Hyaluronan is present in hyaline cartilage, synovial joint fluid, and skin tissue, both dermis and epidermis. Hyaluronan is also suspected of having a role in numerous physiological functions, such as adhesion, development, cell motility, cancer, angiogenesis, and wound healing. Due to the unique physical and biological properties of hyaluronan, it is employed in eye and joint surgery and is being evaluated in other medical procedures.
- The terms “hyaluronan” or “hyaluronic acid” are used in literature to mean acidic polysaccharides with different molecular weights constituted by residues of D-glucuronic and N-acetyl-D-glucosamine acids, which occur naturally in cell surfaces, in the basic extracellular substances of the connective tissue of vertebrates, in the synovial fluid of the joints, in the endobulbar fluid of the eye, in human umbilical cord tissue and in cocks' combs.
- The term “hyaluronic acid” is in fact usually used as meaning a whole series of polysaccharides with alternating residues of D-glucuronic and N-acetyl-D-glucosamine acids with varying molecular weights or even the degraded fractions of the same, and it would therefore seem more correct to use the plural term of “hyaluronic acids”. The singular term will, however, be used all the same in this description; in addition, the abbreviation “HA” will frequently be used in place of this collective term.
- HA plays an important role in the biological organism, as a mechanical support for the cells of many tissues, such as the skin, tendons, muscles and cartilage, it is a main component of the intercellular matrix. HA also plays other important parts in the biological processes, such as the moistening of tissues, and lubrication.
- HA may be extracted from the above mentioned natural tissues, although today it is preferred to prepare it by microbiological methods to minimize the potential risk of transferring infectious agents, and to increase product uniformity, quality and availability.
- HA and its various molecular size fractions and the respective salts thereof have been used as medicaments, especially in treatment of arthropathies, as an auxiliary and/or substitute agent for natural organs and tissues, especially in ophthalmology and cosmetic surgery, and as agents in cosmetic preparations. Products of hyaluronan have also been developed for use in orthopaedics, rheumatology, and dermatology.
- HA may also be used as an additive for various polymeric materials used for sanitary and surgical articles, such as polyurethanes, polyesters etc. with the effect of rendering these materials biocompatible.
- The ASA modification or derivatization is well established in the paper industry where alkyl succinic anhydrides have been used to make paper surfaces (cellulosic) more water resistant (Chen and Woodward, August 1986, Optimizing the emulsification and sizing of alkenyl succinic anhydride, Tappi Journal 95-97). In the food industry 2-octen-1-ylsuccinic anhydride (OSA) modified starches have been used to stabilize oil/water emulsions, e.g., low fat margarines and mayonnaises, (Jarowenko, W. (In: Properties and uses of modified starches, 1986, Ed.: O. Wurzburg) Acetylated starch and miscellaneous organic esters, pp 55-77). Further, the rheological properties of OSA modified starches are very different compared to non-modified starches (Park, Chung, and Yoo, 2004, Effects of octenylsuccinylation on rheological properties of corn starch pastes, Starch 56: 399-406).
- The advantages of the ASA derivatization procedure are, e.g., that the products are non-toxic, the chemicals cheap, and the reaction is a one-step procedure (Trubiano, P C. [In: Properties and uses of modified starches, 1986, Ed.: O. Wurzburg] Succinate and substituted succinate derivatives of starch, pp 131-147; Wurzburg, O B. 1995. Modified starches, In: Food Science and Technology, Vol. 67, New York, pp. 67-97).
- According to earlier studies on starches, both primary and secondary hydroxyl groups react with OSA (Shogren, Viswanathan, Felker, and Gross, 2000, Distribution of octenyl succinate groups in octenyl succinic anhydride modified waxy maize starch, Starch 52:196-204).
- There is a need, particularly in the cosmetics and biomedical industries, for hyaluronic acid based compounds or derivatives that have certain altered characteristics as compared to non-modified HA. Properties of interest are the improved ability to stabilize foam, and the ability to blend with non-hydrophilic materials, such as is used typically in cosmetics products.
- The invention provides amphiphilic HA-derivative products with properties of benefit in cosmetics or biomedical applications. These products bind more strongly to the skin so that they are not so easily washed of. The ASA-HA derivatives are also suitable for use in more advanced cosmetic or biomedical formulations, e.g. in the formation of nano/macro capsules or nano/macro spheres for delivery of active compounds or drugs. ASA-HA derivatives of lower molecular weight (MW) will penetrate the skin more efficiently than non-derivatized HA of comparable MW.
- In the examples herein, hyaluronic acid (HA) was modified with alkyl/aryl succinic anhydrides (ASA) under alkaline conditions (pH>8.0) in water. The resulting products were purified (precipitation or dialysis). These purified products formed partially water-insoluble aggregates in water. A 1% solution was showed to stabilize foam (reduced surface tension+increased interfacial viscosity). 1H NMR spectroscopy confirmed that the chemical structure of the HA “backbone” in the resulting product was unchanged, except for the introduction of ASA half-ester groups up to a degree of substitution (DS) of about 18%.
- Accordingly, in a first aspect, the invention relates to a hyaluronic acid derivative comprising ‘n’ repeating units and having the general structural formula (I) at pH 8-9:
- wherein in at least one repeating unit one or more of R1, R2, R3, R4 comprise an ester bound alkyl/aryl-succinic acid having the general structural formula (II) at pH 8-9, and otherwise R1, R2, R3, R4 are hydroxyl groups, OH:
- wherein at least one of R5, R6, R7, R8 comprises an alkyl- or aryl-group, and otherwise R5, R6,
- R7, R8 are hydrogen atoms, H, and wherein the Oxygen labelled “ester” partakes the ester bond with structure (I).
- In other words, an aspect of the invention relates to a hyaluronic acid derivative, wherein one or more hydroxyl-groups of the hyaluronic acid have been substituted in a reaction with one or more alkyl/aryl-succinic anhydrides (ASA), to form an ester-bond between the hyaluronic acid and the resulting one or more alkyl/aryl-succinic acids.
- In a second aspect, the invention relates to a process of producing a hyaluronic acid derivative, comprising the steps of:
- (a) reacting a hyaluronic acid (HA) with one or more alkyl/aryl-succinic anhydrides (ASA) having the general structural formula shown in (III)
- under alkaline conditions in an aqueous solution, whereby the hyaluronic acid derivative is formed; and
- (b) recovering the hyaluronic acid derivative.
- In a third aspect, the invention relates to a composition comprising a hyaluronic acid derivative as defined in the first aspect and an active ingredient, preferably the active ingredient is a pharmacologically active agent.
- A fourth aspect of the invention relates to a pharmaceutical composition comprising an effective amount of a hyaluronic acid derivative as defined in the first aspect, together with a pharmaceutically acceptable carrier, excipient or diluent.
- A fifth aspect relates to a pharmaceutical composition comprising an effective amount of a hyaluronic acid derivative as defined in the first aspect as a vehicle, together with a pharmacologically active agent.
- A sixth aspect relates to a cosmetic article comprising as an active ingredient an effective amount of a hyaluronic acid derivative as defined in the first aspect or a composition as defined in any of the second, third, or fourth aspects.
- In a seventh aspect, the invention relates to a sanitary, medical or surgical article comprising a hyaluronic acid derivative as defined in the first aspect or a composition as defined in any of the second, third, or fourth aspects, preferably the article is a diaper, a sanitary towel, a surgical sponge, a wound healing sponge, or a part comprised in a band aid or other wound dressing material.
- An important aspect relates to a medicament capsule, microcapsule, nanocapsules, microsphere or nanosphere comprising a hyaluronic acid derivative as defined in the first aspect or a composition as defined in any of the third, fourth, or fifth aspects.
- Final aspects of the invention relate to methods of performing procedures in ophthalmology, in the treatment of osteoarthritis or cancer, hair loss or baldness, of treating a wound, of performing dermal or transdermal administration of a pharmacologically active agent, or dermal administration of a cosmetic, the improvement which comprises the use of a hyaluronic acid derivative as defined in the first aspect, or a composition as defined in any of the second, third, or fourth aspects.
- A number of aspects relate to uses of a hyaluronic acid derivative as defined in any of the first aspects or a composition as defined in any of the third, fourth, or fifth aspects for the manufacture of a medicament for the treatment of osteoarthritis, cancer, the manufacture of a medicament for an ophthalmic treatment, the manufacture of a medicament for the treatment of a wound, the manufacture of a medicament for angiogenesis, the manufacture of a medicament for the treatment of hair loss or baldness, or the manufacture of a moisturizer.
-
FIG. 1 . The schematic ASA modification of HA is illustrated inFIG. 1 . -
FIG. 2 . The partially assigned 1H NMR spectrum of the 100 kDa OSA-HA (14919-033) of examples 5 and 6. -
FIG. 3 . The chemical structure of the ASAs used to modify HA. -
FIG. 4 . Haug's triangle summarizes the relationship between Rg and Mw for different polymer conformations. By plotting radius of gyration (Rg) as against molecular weight (Mw) in double logarithmic scale, one can obtain information about the conformation of the polymer. -
FIG. 5 . Concentration profiles (RI) for the ASA modified LMW HA of example 12 below. -
FIG. 6 . Results of surface tension measurements on LMW OSA-HA using surface tensiometer (Wilhemy plate) as described in example 13. -
FIG. 7 . Shows the emulsification properties of ASA-HA after 24 hours and 8 weeks with ethylhexyl palmitate (cosmetic oil) prepared as described in example 15. -
FIG. 8 . The critical aggregation concentration (CAC) of an OSA-HA derivative was determined by calorimetry as described in Example 18, enthalpy variations (ΔH) in the sample cell were recorded over time as shown inFIG. 8 . -
FIG. 9 . Enthalpy variation (ΔH) of Example 18 was plotted as a function of the OSA-HA concentration in the sample cell, and the CAC of OSA-HA was determined at the break of the curve. Each experiment was repeated three times and the CAC was provided as an averaged value. For example, the CAC of OSA-HA with a degree of substitution (DS) of 16% was 0.45 mg/mL. -
FIG. 10 . Shows the wavelength of maximum emission as a function of OSA-HA (DS=44%) concentration inbuffer 1 as described in Example 19. -
FIG. 11 . Shows the wavelength of maximum emission as a function of OSA-HA (DS=44%) concentration, and that the CAC decreases as the concentration of NaCl increases, as described in Example 20. -
FIG. 12 . Shows the Zeta potential of OSA-HA (DS=44%, 1 mg/mL) in 10-3 M NaCl as determined in Example 21. -
FIG. 13 . Shows a transmission electron micrograph of OSA-HA (DS=44%) polymeric micelles from Example 22, that are spherical in shape and have submicronic dimensions typically from 50 to 200 nm. - “Hyaluronic acid” is defined herein as an unsulphated glycosaminoglycan composed of repeating disaccharide units of N-acetylglucosamine (GlcNAc) and glucuronic acid (GlcUA) linked together by alternating beta-1,4 and beta-1,3 glycosidic bonds. Hyaluronic acid is also known as hyaluronan, hyaluronate, or HA. The terms hyaluronan and hyaluronic acid are used interchangeably herein.
- Rooster combs are a significant commercial source for hyaluronan. Microorganisms are an alternative source. U.S. Pat. No. 4,801,539 discloses a fermentation method for preparing hyaluronic acid involving a strain of Streptococcus zooepidemicus with reported yields of about 3.6 g of hyaluronic acid per liter. European Patent No. EP 0694616 discloses fermentation processes using an improved strain of Streptococcus zooepidemicus with reported yields of about 3.5 g of hyaluronic acid per liter. As disclosed in WO 03/054163 (Novozymes), which is incorporated herein in its entirety, hyaluronic acid or salts thereof may be recombinantly produced, e.g., in a Gram-positive Bacillus host.
- Hyaluronan synthases have been described from vertebrates, bacterial pathogens, and algal viruses (DeAngelis, 1999, Cell. Mol. Life. Sci. 56: 670-682). WO 99/23227 discloses a Group I hyaluronate synthase from Streptococcus equisimilis. WO 99/51265 and WO 00/27437 describe a Group II hyaluronate synthase from Pasturella multocida. Ferretti et al. disclose the hyaluronan synthase operon of Streptococcus pyogenes, which is composed of three genes, hasA, hasB, and hasC, that encode hyaluronate synthase, UDP glucose dehydrogenase, and UDP-glucose pyrophosphorylase, respectively (Proc. Natl. Acad. Sci. USA. 98: 4658-4663 (2001)). WO 99/51265 describes a nucleic acid segment having a coding region for a Streptococcus equisimilis hyaluronan synthase.
- Since the hyaluronan of a recombinant Bacillus cell is expressed directly to the culture medium, a simple process may be used to isolate the hyaluronan from the culture medium. First, the Bacillus cells and cellular debris are physically removed from the culture medium. The culture medium may be diluted first, if desired, to reduce the viscosity of the medium. Many methods are known to those skilled in the art for removing cells from culture medium, such as centrifugation or microfiltration. If desired, the remaining supernatant may then be filtered, such as by ultrafiltration, to concentrate and remove small molecule contaminants from the hyaluronan. Following removal of the cells and cellular debris, a simple precipitation of the hyaluronan from the medium is performed by known mechanisms. Salt, alcohol, or combinations of salt and alcohol may be used to precipitate the hyaluronan from the filtrate. Once reduced to a precipitate, the hyaluronan can be easily isolated from the solution by physical means. The hyaluronan may be dried or concentrated from the filtrate solution by using evaporative techniques known to the art, such as spray drying.
- The first aspect of the invention relates to a hyaluronic acid derivative comprising n repeating units and having the formula (I) at pH 8-9:
- wherein in at least one repeating unit one or more of R1, R2, R3, R4 comprise an ester bound alkyl/aryl-succinic acid having the general structural formula (II) at pH 8-9, and otherwise R1, R2, R3, R4 are hydroxyl groups, OH:
- wherein at least one of R5, R6, R7, R8 comprises an alkyl- or aryl-group, and otherwise R5, R6, R7, R8 are hydrogen atoms, H, and wherein the Oxygen labelled “ester” partakes the ester bond with structure (I).
- In a preferred embodiment of the first aspect, two or more of R1, R2, R3, R4 comprise one or more ester bound alkyl/aryl-succinic acids having the general structural formula (II) at pH 8-9; preferably three or more of R1, R2, R3, R4 comprise one or more ester bound alkyl/aryl-succinic acids having the general structural formula (II) at pH 8-9.
- In another preferred embodiment of the first aspect, at least one of R5, R6, R7, R8 comprises an alkyl-group, preferably at least two of R5, R6, R7, R8 comprise an alkyl-group, more preferably at least three of R5, R6, R7, R8 comprise an alkyl-group; preferably the alkyl-group comprises a C1-C20 alkyl group, preferably propyl, 2-octenyl, 2-nonenyl, 2-dodecenyl, 2-hexadecenyl, or 2-octadecenyl.
- Yet another preferred embodiment relates to the HA derivative of the first aspect, wherein at least one of R5, R6, R7, R8 comprises an aryl-group, preferably at least two of R5, R6, R7, R8 comprise an aryl-group, more preferably at least three of R5, R6, R7, R8 comprise an aryl-group; and preferably the aryl-group is phenyl.
- It is preferred that R5, R6, R7, R8 comprise two or more different alkyl- and/or aryl-groups, preferably chosen from propyl, 2-octenyl, 2-nonenyl, 2-dodecenyl, 2-hexadecenyl, 2-octadecenyl, and phenyl.
- The level of hyaluronic acid may be determined according to the modified carbazole method (Bitter and Muir, 1962, Anal Biochem. 4: 330-334). Moreover, the average molecular weight of the hyaluronic acid may be determined using standard methods in the art, such as those described by Ueno et al., 1988, Chem. Pharm. Bull. 36: 4971-4975; Wyatt, 1993, Anal. Chim. Acta 272: 1-40; and Wyatt Technologies, 1999, “Light Scattering University DAWN Course Manual” and “DAWN EOS Manual” Wyatt Technology Corporation, Santa Barbara, Calif.
- In a preferred embodiment, the hyaluronic acid derivatives obtained by the methods of the present invention has a molecular weight of about 800 to about 10,000,000 Da. In a more preferred embodiment, the hyaluronic acid derivatives obtained by the methods of the present invention has a molecular weight of about 1,000 to about 9,000,000 Da; about 2,000 to about 10,000,000 Da; about 4,000 to about 10,000,000 Da; about 8,000 to about 10,000,000 Da; about 10,000 to about 10,000,000 Da; or about 25,000 to about 5,000,000 Da. In an even more preferred embodiment, the hyaluronic acid derivatives obtained by the methods of the present invention has a molecular weight of about 50,000 to about 3,000,000 Da.
- Another preferred embodiment relates to the product of the first aspect, wherein the hyaluronic acid or salt thereof has a molecular weight in the range of between 300,000 and 3,000,000; preferably in the range of between 400,000 and 2,500,000; more preferably in the range of between 500,000 and 2,000,000; and most preferably in the range of between 600,000 and 1,800,000 Da.
- Where recombinantly produced hyaluronic acid or salt thereof is used in the methods of the invention to manufacture the products or compositions of the invention, it may be advantageous for some applications to first reduce the average molecular weight of the hyaluronic acid or derivative or salts thereof. For instance, it has been stated by several manufacturers of so-called low-molecular weight fractions of hyaluronic acid, that it is capable of penetrating the skin barrier to reestablish the natural content of hyaluronic acid in the skin, therefore such fractions are particularly suitable for cosmetic compositions sold as anti-skin-ageing and anti-wrinkle agents. For food applications, low MW hyaluronic acid has been shown to penetrate the gastrointestinal barrier, thereby increasing its bioavailability. Finally, low MW hyaluronic acid exhibits anti-inflammatory effect and have potential applications in the treatment of inflammatory diseases. A reduction of the average molecular weight of a hyaluronic acid or derivative or salt thereof may be achieved by standard methods in the art, such as, simple heat treatment, enzymatic degradation, ultrasound sonication, or acid hydrolysis. See, e.g., U.S. Pat. No. 6,020,484, which describes an ultrasonication technique of HA including NaOCl as additive, and Miyazaki et al., 2001, Polymer Degradation and Stability 74: 77-85.
- Accordingly, a preferred embodiment relates to the HA derivative of the invention, wherein the hyaluronic acid or derivative or salt thereof has a low average molecular weight in the range of between 800 and 10,000,000 Da; preferably in the range of between 10,000 and 1,500,000 Da; preferably in the range of between 10,000 and 50,000 Da; or preferably in the range of between 50,000 and 500,000 Da; even more preferably in the range of between 80,000 and 300,000 Da.
- DS was determined by 1H NMR spectroscopy (10 mg/ml, D2O, 80° C., 128 scans, 400 MHz) according to example 6 below, wherein the peaks from the OSA group were assigned by use of a 2D-NMR (gCOSY). The DS was then calculated by comparing the intensity of the vinyl protons of OSA (5.4 and 5.6 ppm) with that of the acetyl protons (2.0 ppm).
- In a preferred embodiment the HA derivative of the first aspect has a Degree of Substitution (DS) in the range of 0.1-100%, preferably 1-90%, more preferably 2-80%, still more preferably 4-70%, even more preferably 8-60%, or 10-50%, 14-40%, 16-30%, or most preferably in the range of 18-25%.
- In a preferred embodiment of the invention, the one or more alkyl/aryl-succinic anhydrides (ASA) have the general structural formula (III):
- In one preferred embodiment, at least one of R5, R6, R7, R8 comprises an alkyl-group, more preferably at least two of R5, R6, R7, R8 comprise an alkyl-group, even more preferably at least three of R5, R6, R7, R8 comprise an alkyl-group; and preferably the alkyl-group comprises a C1-C20 alkyl group, preferably propyl, 2-octenyl, 2-nonenyl, 2-dodecenyl, 2-hexadecenyl, or 2-octadecenyl.
- In another preferred embodiment, at least one of R5, R6, R7, R8 comprises an aryl-group, preferably at least two of R5, R6, R7, R8 comprise an aryl-group, more preferably at least three of R5, R6, R7, R8 comprise an aryl-group, which preferably comprises phenyl.
- In yet another preferred embodiment R5, R6, R7, R8 comprises two or more different alkyl- and/or aryl-groups, preferably chosen from propyl, 2-octenyl, 2-nonenyl, 2-dodecenyl, 2-hexadecenyl, 2-octadecenyl, and phenyl.
- In still another preferred embodiment, the one or more ASA comprise any of the structural formulae shown in
FIG. 3 . - In the methods of the present invention recombinantly produced HA may be used that is produced by a process, wherein the HA-producing host cells are cultivated in a nutrient medium suitable for production of the hyaluronic acid using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the enzymes involved in hyaluronic acid synthesis to be expressed and the hyaluronic acid to be isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art. Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection). The secreted hyaluronic acid can be recovered directly from the medium.
- The resulting hyaluronic acid may be isolated by methods known in the art. For example, the hyaluronic acid may be isolated from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The isolated hyaluronic acid may then be further purified by a variety of procedures known in the art including, but not limited to, chromatography (e.g., ion exchange, affinity, hydrophobic, chromatofocusing, and size exclusion), electrophoretic procedures (e.g., preparative isoelectric focusing), differential solubility (e.g., ammonium sulfate precipitation), or extraction (see, e.g., Protein Purification, J.-C. Janson and Lars Ryden, editors, VCH Publishers, New York, 1989).
- A preferred embodiment relates to where the hyaluronic acid or salt thereof is recombinantly produced, preferably by a Gram-positive bacterium or host cell, more preferably by a bacterium of the genus Bacillus.
- The host cell may be any Bacillus cell suitable for recombinant production of hyaluronic acid. The Bacillus host cell may be a wild-type Bacillus cell or a mutant thereof. Bacillus cells useful in the practice of the present invention include, but are not limited to, Bacillus agaraderhens, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis, and Bacillus thuringiensis cells. Mutant Bacillus subtilis cells particularly adapted for recombinant expression are described in WO 98/22598. Non-encapsulating Bacillus cells are particularly useful in the present invention.
- In a preferred embodiment, the Bacillus host cell is a Bacillus amyloliquefaciens, Bacillus clausii, Bacillus lentus, Bacillus licheniformis, Bacillus stearothermophilus or Bacillus subtilis cell. In a more preferred embodiment, the Bacillus cell is a Bacillus amyloliquefaciens cell. In another more preferred embodiment, the Bacillus cell is a Bacillus clausii cell. In another more preferred embodiment, the Bacillus cell is a Bacillus lentus cell. In another more preferred embodiment, the Bacillus cell is a Bacillus licheniformis cell. In another more preferred embodiment, the Bacillus cell is a Bacillus subtilis cell. In a most preferred embodiment, the Bacillus host cell is Bacillus subtilis A164Δ5 (see U.S. Pat. No. 5,891,701) or Bacillus subtilis 168Δ4.
- Transformation of the Bacillus host cell with a nucleic acid construct of the present invention may, for instance, be effected by protoplast transformation (see, e.g., Chang and Cohen, 1979, Molecular General Genetics 168: 111-115), by using competent cells (see, e.g., Young and Spizizen, 1961, Journal of Bacteriology 81: 823-829, or Dubnau and Davidoff-Abelson, 1971, Journal of Molecular Biology 56: 209-221), by electroporation (see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), or by conjugation (see, e.g., Koehler and Thorne, 1987, Journal of Bacteriology 169: 5271-5278).
- A preferred embodiment relates to a hyaluronic acid derivative of the first aspect, which comprises an inorganic salt of hyaluronic acid, preferably sodium hyaluronate, potassium hyaluronate, ammonium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, or cobalt hyaluronate.
- The preparation of a crosslinked HA or salt thereof, which is prepared by crosslinking HA with a polyfunctional epoxy compound is disclosed in EP 0161887 B1. Total or partial crosslinked esters of HA with an aliphatic alcohol, and salts of such partial esters with inorganic or organic bases, are disclosed in U.S. Pat. No. 4,957,744. Other ways of cross-linking HA are disclosed in U.S. Pat. Nos. 5,616,568, 5,652,347, and 5,874,417.
- Crosslinked HA may also be prepared by treating HA with boric acid, as follows: Dried sodium hyaluronate (Na-HA, 203 mg), recombinantly produced in a Bacillus subtilis by fermentation (WO 03/054163; Novozymes), was dissolved into 0.2 M NaOH to give a 4% solution. Boric acid (35 mg (approx. 1 equivalent of HA disaccharide) was added and the sample was stirred at room temperature for 1.5 h, and then stored at 5° C. for approx. 2.5 days. A control sample was prepared in parallel exactly as described above, but without boric acid. The viscosity of the resulting HA-borate hydrogel was measured at 25° C. using a Carrimed CSL controlled stress rheometer (cone geometry: 6 cm, 2°). The viscosity depended on the shear rate and increased at least 4-fold (from 4.2- to 8.4 fold) in the HA-borate hydrogel as compared to the control sample, indicating formation of a cross-linked network. New peaks at 1200 and 945 cm−1 were observed on the FT-IR spectrum of the HA-borate hydrogel, when compared to a standard spectrum of Na-HA, corresponding to the presence of newly formed borate esters in the crosslinked HA-borate hydrogel.
- Accordingly, a preferred embodiment relates to the HA derivative of the first aspect, which comprises crosslinked hyaluronic acid or salt thereof, preferably the hyaluronic acid is crosslinked with boric acid, and more preferably the crosslinked hyaluronic acid comprises borate esters.
- A preferred HA derivative of the first aspect has a particle size the 50 percentile of which, D50, is between 10 and 1,000 microns, preferably between 100 and 1,000 microns, more preferably between 150 and 900 microns, and even more preferably between 200 and 800 microns, as determined by laser diffraction measurement of the particles suspended in isopropanol.
- In a preferred embodiment, the polydispersity of a HA derivative of the first aspect is measured as the SPAN value, which is calculated according to the following formula: SPAN=(D90-D10)/D50, and the SPAN value is between 1.0 and 2.5; preferably the SPAN value is between 1.2 and 2.2; more preferably the SPAN value is between 1.5 and 1.9; and most preferably the SPAN value is between 1.6 and 1.8.
- As shown in the examples below, the present invention provides ASA-HA derivatives that are capable of forming micro- or nanoparticles, or micro- or nanocapsules. Such particles or capsules, or compositions comprising these, may of use in a large number of commercial and scientific applications, such as in cosmetics or in general drug-delivery.
- In a preferred embodiment, the compositions comprising a HA derivative of the invention may also comprise other ingredients, preferably one or more active ingredients, preferably one or more pharmacologically active substances, and also preferably a water-soluble excipient, such as lactose.
- Non-limiting examples of an active ingredient or pharmacologically active substance which may be used in the present invention include protein and/or peptide drugs, such as, human growth hormone, bovine growth hormone, porcine growth hormone, growth hormone releasing hormone/peptide, granulocyte-colony stimulating factor, granulocyte macrophage-colony stimulating factor, macrophage-colony stimulating factor, erythropoietin, bone morphogenic protein, interferon or derivative thereof, insulin or derivative thereof, atriopeptin-III, monoclonal antibody, tumor necrosis factor, macrophage activating factor, interleukin, tumor degenerating factor, insulin-like growth factor, epidermal growth factor, tissue plasminogen activator, factor VII, factor VIII, and urokinase.
- A water-soluble excipient may be included for the purpose of stabilizing the active ingredient(s), such excipient may include a protein, e.g., albumin or gelatin; an amino acid, such as glycine, alanine, glutamic acid, arginine, lysine and a salt thereof; carbohydrate such as glucose, lactose, xylose, galactose, fructose, maltose, saccharose, dextran, mannitol, sorbitol, trehalose and chondroitin sulphate; an inorganic salt such as phosphate; a surfactant such as TWEEN® (ICI), poly ethylene glycol, and a mixture thereof. The excipient or stabilizer may be used in an amount ranging from 0.001 to 99% by weight of the product.
- Several aspects of the invention relate to various compositions and pharmaceutical comprising, among other constituents, an effective amount of the product as defined in the first aspect, and an active ingredient, preferably the active ingredient is a pharmacologically active agent; a pharmaceutically acceptable carrier, excipient or diluent, preferably a water-soluble excipient, and most preferably lactose.
- In addition, aspects of the invention relate to articles comprising a HA derivative as defined in the first aspect or a composition as defined in the aspects and embodiments above, e.g., a cosmetic article, a sanitary article, a medical or surgical article. In a final aspect the invention relates to a medicament capsule or microcapsule comprising a product as defined in the first aspect or a composition as defined in other aspects and embodiments of the invention.
- High molecular weight (High-MW) Hyaluronic acid (HA):
-
- (batch MAG 30014)
- (batch MAF 145 SD)
Low molecular weight (Low-MW) HA: - 100 kDa (batch 14919-021)
- kDa (batch 14919-032)
- 23 kDa
- 14 kDa
-
-
- cis/trans-2-octen-1-ylsuccinic anhydride (OSA), Aldrich Chemical Company (d.: 1.000, MW 220.27, 97% purity).
- Phenylsuccinic anhydride (PhSA) (Dry powder, MW 176.17).
- Nonenylsuccinic anhydride (NSA) Aldrich Chemical Company (d.: 1.032, MW 224.30, 95+% purity, 1JS38, 246-00198-1).
- Dodecenylsuccinic anhydride (DSA) (d.: 1.01, MW 266.38, 1JS38, 246-00168).
- Tetrapropylsuccinic anhydride (TpSA).
- Hexadecenyl succinic anhydride (HDSA)
- Octadecenyl succinic anhydride (ODSA)
- Mixture (50:50) of ODSA and HDSA
Ethanol 96%, denaturated.
- Milli-Q® ultrapure water (Millipore).
Dialysis tubes of regenerated cellulose with a molecular weight cutoff of 12-14 kDa, Spectr/Por™ (Spectrum Medical Industries).
Ultrafiltration membranes (MWCO 10 kDa and 3 kDa). - HA (batch MAF 145 SD, 1.42 g) was dissolved overnight at room temperature in Milli-Q water (200 mL) before adjusting pH to 9.0 with 4 M NaOH. OSA (1 mL, 4.54 mmol) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. 20 mL saturated NaHCO3 was added to buffer the reaction. After the reaction, the pH was adjusted to 6.8 with 1 M HCl. The product was recovered by ethanol precipitation by adding 96% ethanol (4 volumes) to give a final concentration of 80% v/v. The precipitate was recovered by centrifugation (3000 rpm, 15 min and 4° C.). The pellet was washed with 96% ethanol before re-dissolving in MQ water and freeze drying.
- To each of three 50 mL solutions of Milli-Q water, HA (batch MAF 145 SD, 1.13 g) was added and left to dissolve overnight at room temperature. The pH was adjusted to 11 with 4 M NaOH. Different amounts of OSA (1.10 mL (5.23 mmol), 0.505 mL (2.62 mmol), 0.110 mL (0.52 mmol)) was added to each of the three solutions under strong agitation. The solutions were left to react on strong agitation (approx 600 rpm) for 21 hours at ambient temperature. All samples had a pH of around 4-5 after the reaction. The product was recovered by ethanol precipitation by adding 96% ethanol (4 volumes) to give a final concentration of 80% v/v. The precipitate was recovered by centrifugation (3000 rpm, 15 min and 4° C.). The pellet was washed with 96% ethanol before re-dissolving in MQ water and freeze drying.
- HA (batch MAF 145 SD, 0.75 g) was dissolved overnight at room temperature in Milli-Q water (150 mL) before adjusting pH to 9.0. OSA (1.42 mL, 6.25 mmol) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 9-11 by use of a pH stat (adding 1 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized.
- HA (batch MAG 30014, 0.75 g) was dissolved overnight at room temperature in Milli-Q water (150 mL) before adjusting pH to 8.5. OSA (1.42 mL, 6.25 mmol) was added under strong agitation. The solution was left on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 9-11 by use of a pH stat (adding 1 M NaOH). pH was adjusted to 6.5 by use of 1 M HCl. The product was dialyzed 3×3 h against 0.2 M NaOH, and 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized.
- Low-MW HA (30 or 100 kDa, 2.5 g) was dissolved overnight at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of OSA (3.35 mL, HA:OSA ratio 1:1) or 1/10 of the molar concentration of HA (0.35 mL, HA:OSA ratio 10:1) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 1 M or 0.5 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized.
- Molecular weight
- 100 kDa OSA-HA from example 5 was analyzed using SEC-MALLS-VISC (mobile phase: 150 mM NaCl, 50 mM NaH2PO4, pH 7.0, 0.5 ml/min, injected volume: 0.5 ml). Columns used: PL aquagel OH-40/OH-50/0H60. System: Waters Alliance HPLC system Waters 2410 RI detector and Wyatt MALLS detector. The data was processed using the ASTRA V software from Wyatt Technology Corp.
- DS was determined by 1H NMR spectroscopy (10 mg/ml, D2O, 80° C., 128 scans, 400 MHz). The peaks from the OSA group were assigned by use of a 2D-NMR (gCOSY).
- During all experiments on high-MW HA the observations have been the same: OSA forms amber-coloured oil drops that gradually divide into smaller drops because of the agitation. At the end of the reaction, the solution was white and opaque like milk which can be interpreted as formation of micelles or micro-scale aggregates/droplets. Even after purification by precipitation or dialysis, where the excess of OSA is removed, this phenomenon was still observed to different extents.
- During the first experiments on high-MW HA, precipitation in 80% ethanol was used to remove the surplus/by-product of the OSA modification. However, due to problems with getting the product to precipitate completely, dialysis against Milli-Q water was chosen as a better method.
- The initial preparations of high-MW OSA-HA all showed changes in solution properties. One example is that they all stabilized foam very efficiently for several hours; this was simply tested by shaking a 1% solution followed by visual inspection. Another observation during the first experiments was that the pH value of the solution declines gradually during the reaction. Therefore, it was necessary to either buffer the system, e.g., with NaCO3 or by use of a pH stat. It is important that the pH value remains above 8.0 for the reaction to proceed, and below 9.0 to avoid removing the OSA groups by hydrolysis.
- NMR spectroscopy was attempted on the high-MW OSA-HA products, but because of solubility problems only some very weak peaks of OSA and HA were observed, and no DS could be determined. In all cases the yields were close or slightly higher than the amount of starting material HA (determined by weighing the lyophilized products).
- Four separate experiments were performed and are summarised in Table 1 together with DS from 1H-NMR spectroscopy and yields. The DS is calculated by comparing the intensity of the vinyl protons of OSA (5.4 and 5.6 ppm) with that of the acetyl protons (2.0 ppm).
- The 1H NMR spectrum of the 30 kDa OSA-HA was elucidated by 2D NMR spectroscopy (gCOSY), and the partially assigned peaks are given in
FIG. 2 , showing the 1H NMR spectrum of the 100 kDa OSA-HA (14919-033). - Conclusively, in these experiments both high- and low-MW hyaluronic acid was successfully modified with 2-octen-1-yl succinic anhydride (OSA).
-
TABLE 1 Results from the preparation of Low-MW OSA-HA. Batch 14658-129 14658-131 14658-133 14919-033 14919-038 HA:OSA 1:1 1:1 10:1 1:1 10:1 MW 30 kDa 30 kDa 30 kDa 100 kDa 100 kDa Yield 2.1 g 2.1 g 2.8 g 2.7 g 2.0 g DS 11.5% 12% 2.6% 18.8% 1.6% - Low-MW HA (14 kDa, 2.5 g) was dissolved at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of OSA (3.35 mL, HA:OSA ratio 1:1) or 1/10 of the molar concentration of HA (0.34 mL, HA:OSA ratio 10:1) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 0.5 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized. Yields after purification and freeze-drying were 2.1 g and 2.1 g, respectively. DS were determined as described in Example 6 to 11.5% and 2.6%, respectively.
- Low-MW HA (14 kDa, 2.5 g) was dissolved at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of PhSA (2.8 g, HA:PhSA ratio 1:1) or 1/10 of the molar concentration of HA (0.28 g, HA:PhSA ratio 10:1) was added gradually under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 0.5 M NaOH). The product was dialyzed 3×3 hours against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized. Yields after purification and freeze-drying were 2.5 g and 2.4 g, respectively. DS were determined as described in Example 6 to 15.1% and 2.6%, respectively.
- Low-MW HA (14 kDa, 2.5 g) was dissolved at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of NSA (3.55 mL, HA:NSA ratio 1:1) or 1/10 of the molar concentration of HA (0.35 mL, HA:NSA ratio 10:1) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 0.5 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized. Yields after purification and freeze-drying were 2.4 g and 2.2 g, respectively. DS were determined as described in Example 6 to 11.4% and 2.1%, respectively.
- Low-MW HA (14 kDa, 2.5 g) was dissolved at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of DSA (4.20 mL, HA:DSA ratio 1:1) or 1/10 of the molar concentration of HA (0.42 mL, HA:DSA ratio 10:1) was added under strong agitation. The solution was left to react on strong agitation (approx 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 0.5 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized. Yields after purification and freeze-drying were 2.2 g and 2.2 g, respectively. DS were determined as described in Example 6 to 2.2% and 1.7%, respectively.
- Low-MW HA (14 kDa, 2.5 g) was dissolved at room temperature in Milli-Q water (50 mL) before adjusting pH to 8.5. Equimolar amounts of TpA (3.25 mL, HA:TpSA ratio 1:1) or 1/10 of the molar concentration of HA (0.33 mL, HA:TpSA ratio 10:1) was added under strong agitation. The solution was left to react on strong agitation (approximately 600 rpm) for 16 hours at ambient temperature. The pH was maintained around 8.5-9.0 by use of a pH stat (adding 0.5 M NaOH). The product was dialyzed 3×3 h against MQ water (4° C., 7 L, MWCO 12-14,000 Da), frozen and lyophilized.
- Eleven separate experiments were performed modifying LMW HA (14 kDa) with five different ASAs (see
FIG. 3 for the ASA names, structures, and abbreviations) at two different HA:ASA molar ratios (1:1 and 10:1). The resulting products were purifies by dialysis to remove excess reagent and byproducts. The degree of substitution (DS) was determined on monomer basis by 1H NMR spectroscopy. Yield was determined gravimetrically the freeze dried samples. Molecular weight was determined by SEC-MALLS-VISC. To avoid material getting stuck on the GPC columns the temperature was adjusted from 4° C. to 15° C. in the auto-injector. All results from the analyses are summarised in Table 2. -
TABLE 2 Results from the preparation and characterisation of ASA modified LMW HA (14 kDa starting material). Degree of Molecular Conformational Ratio substitution weight, plot factor; v Sample ASA (ASA:HA) (%) Yield (g) Mw (kDa) (Rg~Mv) 14658-142 OSA 1:1.25 9.7 2.70 17.0 0.35 ± 0.04 14658-144 OSA 1:0.125 3.5 2.37 14.2 0.56 ± 0.05 14658-148 PhSA 1:1.25 1.8 2.19 13.8 0.62 ± 0.04 15286-017a PhSA 1:1.25 15 1.80 15.9 0.67 ± 0.02 14658-150 PhSA 1:0.125 3.4 2.19 14.3 0.58 ± 0.02 15286-010 NSA 1:1.25 11 2.35 21.1c 0.04 ± 0.02 15286-012 NSA 1:0.125 2.1 2.24 14.4 0.20 ± 0.01 15286-014b DSA 1:1.25 2.2 2.20 14.1 0.61 ± 0.02 15286-020b DSA 1:0.125 1.7 2.24 14.0 0.49 ± 0.02 15286-028 TpSA 1:1.25 * * * * 15286-037 TpSA 1:0.125 * * * * aRepeated and downscaled version of 14658-148. PhSA was added in small portions instead of all at once. bProblems with DSA - too thick to disperse efficiently with normal stirring; an oil phase was formed during dialysis that had to be removed by pipette and discarded. cBimodal distribution; peak 1: 15.5 kDa, peak 2: 27.2 kDa (*): Currently being analyzed. - As can be seen from the results summarized in table 2, the derivatization reaction runs smoother when a lower DS is the desired outcome. The obtained DS's are quite similar for all the different ASAs, except for the PhSA which apparently is very instable in water, resulting in a low DS value (2.19) for sample 14658-148. The experiment was repeated where the PhSA powder was added gradually to the HA-solution. This resulted in a DS of 15% (15286-017), showing that gradual addition of the ASA could be a way of increasing the substitution on HA.
- For the LMW HA modified with DSA, the DS values are also quite low at the higher ASA:HA ratio. This is probably because of the high viscosity of the DSA phase. In addition, the droplets of non-reacted ASA could still be seen after dialysis and freeze drying. This had to be removed manually by a pipette.
- Probably, the DS and purity of the DSA-HA can be improved by increasing the temperature during the reaction combined with stronger agitation. Adding the DSA gradually may also increase the amount of DSA reacted with HA. The TpSA samples (15286-037 and 15286-028) have not yet been analyzed yet.
- By plotting radius of gyration (Rg) as against molecular weight (Mw) in double logarithmic scale, one can obtain information about the conformation of the polymer. The relationship Rg and Mw is summarised in Haug's triangle (
FIG. 4 ). - Conformational plots were made for all of the different ASA-HA, and the results are given in Table 2. Most of the samples show factors similar to that of random coils, which is also the conformation of non-modified HA. The only exceptions, are the highly modified OSA-HA's, which have a conformation similar to that of a sphere, and the NSA-HA's that also show very low conformational factors (0.20 and 0.04) indicating aggregation or impaired column separation, perhaps because of interactions with the column material. Similarly, looking at the concentration profile (RI-signal) from the SEC-MALLS-VISC analysis (See
FIG. 5 ), there is an aggregation peak at an earlier elution time, at approximately 35 minutes, for sample 15286-010 (11% NSA). This aggregation phenomenon is also indicated by the slight increase in the apparent MW (Table 2) for samples 14658-142 (9.7% OSA modified HA) and 15286-010 (11% modified NSA-HA). - In conclusion, low MW hyaluronic acid (14 kDa) was successfully modified with diverse aryl/alkyl succinic anhydrides. High DS products of OSA-HA and NSA-HA show some aggregation tendencies and changes in conformation, probably caused by hydrophobic interactions.
- Solutions of 14 kDa OSA-HA (DS=9.7%, batch 14658-142) and unmodified LMW HA (30 kDa) were prepared in MQ-water according to the concentrations given in Table 3. The samples were analyzed by surface tension measurements using a surface tensiometer (Wilhelmy plate). Surface tension of the solvent (water) was determined to 72 mN/m with the same method. Results of the surface tension measurements are given in
FIG. 6 and summarized in Table 3. As can be seen, the surface tension decreases with increasing concentration of OSA-HA. Comparing with the pure solvent (MQ-water) and LMW-HA (30 kDa), the surface tension is much lower for the HA derivatives. - Further, it can be seen that the surface tension continues to decrease in a time dependant manner for the OSA-HAs. This can be explained by the fact that OSA-HA works as a high MW surfactant, using long time to diffuse to the surface of the solution (since diffusion speed is inversely proportional to MW). More surface active polymer at the surface gives lower surface tension. This time dependence is also a further proof of the OSA moieties actually being covalently bound to the HA, and not only co-existing with HA in the solution.
- This further implies that OSA modification of HA is not rendering it hydrophobic, but amphiphilic. These properties can potentially be exploited in systems where lower surface tension is needed (e.g., local ophthalmic) or where emulsifying properties are needed to stabilize emulsions or foams in cosmetics or pharmaceutical formulations.
-
TABLE 3 Surface tension measurements of OSA-HA solutions in MQ-water at 3200 seconds. Concentration LMW OSA-HA LMW HA (%) (14 kDa, DS = 9.7%) (30 kDa) 0.01 64 mN/m — 0.1 52 mN/m 64 mN/m 1.0 40 mN/m — - HA (4 g, MAG30021) was dissolved overnight in 400 mL MQ water. Solutions were kept at room temperature (25° C.) or heated to 60° C. before Na2CO3 (2 g) was added under shear (ULTRA-TURRAX 24 000 min−1, 5 min). Then the ASA was added according to reaction scheme presented in Table 4 and mixed under strong shear (ULTRA-TURRAX 24 000 min−1, 5 min). The resulting emulsions were left to react for 6 hours at the given temperature (Table 4), then removed to room temperature over night. The pH was adjusted to neutrality before the products were purified by ultrafiltration (
MWCO 10 000) until conductivity was below 15 μSi/cm. The products were frozen and lyophilized. NMR spectroscopy confirmed that all the products were modified. Sample all samples gave turbid solutions in 0.1 M NaCl at 1% w/v concentration. -
TABLE 4 Reaction Scheme for preparation of high MW ASA-derivatives ASA:HA Reaction temp. Sample ID ASA Carbon chain Molar ratio [° C.] 1 OSA C8 1:1 25 2 OSA C8 1:10 25 3 ODSA C18 1:1 60 4 ODSA C18 1:10 60 5 HDSA C16 1:1 60 6 DSA C12 1:1 60 7 DSA C12 1:10 60 ODSA: Octadecenyl succinic anhydride, HDSA: Hexadecenyl succinic anhydride, DSA: Dodecenyl succinic anhydride. - The ASA-HAs no. 1, 3, 4, 5, 6, 7, and non-modified HA (MAG30014), prepared in example 14 were formulated with three cosmetic oils (mineral oil, diethylhexyl carbonate and ethylhexyl palmitate) according to the following recipe:
-
- 1. 6 mL oil was added to 14 mL aqueous solution of 0.1 M NaCl and 0.29% ASA-HA
- 2. The solution was mixed under strong shear for 25 seconds (ULTRA-TURRAX at 24 000 min−1).
- 3. The emulsions were left at room temperature in the dark for 8 weeks, being evaluated visually after 24 hour and 8 weeks for stability.
- All derivatives showed increased emulsion stability compared to the control and the non-modified starting HA (see
FIG. 7 for samples after 24 hour and 8 weeks for ethylhexyl palmitate). This shows that ASA-HA can be used as emulsifiers in cosmetics or advanced drug delivery systems based on emulsions. - Low MW ASA HA was prepared as described in example 15 for high MW HA, the only difference being that the starting concentrations of HA (23 kDa) were 2% w/v. All samples in Table 5 were prepared at 60° C. and purified by ultrafiltration (MWCO 3000 kDa) and lyophilization. The DS were determined by 1H NMR spectroscopy as described in example 6.
-
TABLE 5 Reaction scheme for preparation of low MW HA derivatives with longer alkyl chains. Ratio DS (NMR Sample # ASA (HA/ASA) spectroscopy) Yield 15286-109-1 DSA 1:1.25 2.12% 4.09 g (HA: 23 kDa) 15286-109-2 DSA 1:0.125 Confirmed 3.44 g (HA: 23 kDa) modified 15286-118-1 HDSA/ODSA 1:1.25 13.14% 6.73 g (HA: 23 kDa) (50:50) 15286-118-2 HDSA/ODSA 1:0.125 1.06% 3.84 g (HA: 23 kDa) (50:50) 15286-120-1 ODSA 1:1.25 12.94% 6.15 g (HA: 23 kDa) 15286-120-2 ODSA 1:0.125 0.08% 2.96 g (HA: 23 kDa) - PhSA-HA (14 kDa) (10 mg/ml) in aqueous solution has been shown to degrade much faster than non-modified HA in the presence of hydroxyl radicals (generated by Cu2+/H2O2) followed by both streak camera observations and light scattering (DLS/SLS) studies. This shows that PhSA-HA has a potential as a free-radical scavenging agent for potential use in cosmetic formulations.
- The critical aggregation concentration (CAC) of an OSA-HA derivative was determined by calorimetry using an isothermal titration calorimeter VP-ITC (Microcal LLC, USA). A concentrated solution of OSA-HA (0.294 mL, 15 mg/mL in distilled water) was used to titrate distilled water (1.4615 mL) in the calorimeter sample cell. A solution of OSA-HA (2 μL, 15 mg/mL) was injected every 300 seconds, and enthalpy variations (ΔH) in the sample cell were recorded over time as shown in
FIG. 8 . - ΔH was plotted as a function of the OSA-HA concentration in the sample cell, and the CAC of OSA-HA was determined at the break of the curve. Each experiment was repeated three times and the CAC was provided as an averaged value. For example, the CAC of OSA-HA with a degree of substitution (DS) of 16% was 0.45 mg/mL (
FIG. 9 ). - This study confirmed the existence of associative properties of OSA-HA. Moreover, it indicated the potential formulation of these derivatives into micelles and/or micro-/nanoparticles, making them suitable for use in the encapsulation and delivery of hydrophobic compounds such as hydrophobic cosmetic bioactives and drugs.
- The CAC of OSA-HA with a degree of substitution of 44% was determined by fluorescence spectroscopy using a spectrofluorometer (FluoroMax, Spex, United States) thermostated with a water bath (Julabo F10, Merck, United States). Nile Red was employed as the fluorescent probe. Fluorescence was measured on a range of OSA-HA solutions (Table 6) prepared in different phosphate buffers (Table 7).
-
TABLE 6 OSA-HA solutions Concentration Solution OSA-HA (mg/mL) 1 0.0001 2 0.0002 3 0.0006 4 0.001 5 0.002 6 0.006 7 0.01 8 0.02 9 0.06 10 0.1 11 0.2 12 0.6 13 1.0 -
TABLE 7 Phosphate buffers Concentration Concentration Buffer NaCl (M) NaH2PO4 (M) 1 0.15 0.01 2 0.50 0.01 3 1.00 0.01 4 1.50 0.01 - Nile Red (3.184 mg) was dissolved in a mixture of THF and acetone (50/50, 10 mL). This solution (10 μL) was incubated with each OSA-HA solution (10 mL) under stirring, overnight, in the dark and at room temperature. Each solution was analyzed at 25° C. at an excitation wavelength of 543 nm whereas emission spectra were recorded from 580 to 700 nm. The excitation slit was set to 1 and the emission slit was adjusted for each solution. The intensity of the fluorescence emission (I) was plotted as a function of the wavelength (λ).
- The wavelength corresponding to the maximum intensity (λ max) was determined by fitting the curve I vs. λ with a polynomial function of
order 6. Each λ max value was the average of three measurements. - In order to determine the CAC, λ max was plotted as a function of the polymer concentration (C). The CAC was deduced at the inflexion point of the curve λ max vs. C (
FIG. 10 ). - In
FIG. 10 the CAC of OSA-HA (DS=44%) was somewhere between 0.003 and 0.004 mg/mL. This phenomenon was not observed for unmodified HA. Indeed fluorescence could not be detected at any HA concentrations which means that it was not possible to solubilize Nile Red in HA solutions. This evidences the presence of polymeric assemblies in OSA-HA solutions. - The same experimental set-up as the one described in the previous example was used to study the influence of salt concentration on the value of CAC of OSA-HA (DS=44%). The results are shown in table 8 below as well as in
FIG. 11 . -
TABLE 8 Concentration Onset of CAC Extend of the CAC of NaCl (M) (mg/mL) transition (mg/mL) (mg/mL) 0.15 0.002 0.002-0.006 0.003-0.004 0.5 0.0006 0.0006-0.006 0.003 1.0 0.0006 0.0006-0.006 0.002 1.5 0.0002 0.0002-0.002 0.0006-0.0007 - The zeta potential of OSA-HA (DS=44%) polymeric micelles was determined by capillary electrophoresis (Zetasizer 3000HS, Malvern, United Kingdom) coupled to a Doppler laser interferometer. Measurements were recorded at 25° C. OSA-HA was dissolved in 10-3 M NaCl (at a concentration of 1 mg/mL) prior to the measurement. The zeta potential of OSA-HA (DS=44%, 1 mg/mL in 10-3 M NaCl) was evaluated to approximately −25 mV (
FIG. 12 ). - Microscopic observations of OSA-HA (DS=44%) polymeric micelles were made with a transmission electron microscope (EM 410, Philips, The Netherlands). Samples were deposited on ionised carbon coated copper grids and stained with an aqueous uranyl acetate solution (2%). Microscopic snapshots clearly showed that the OSA-HA polymeric micelles are spherical in shape and have submicronic dimensions typically from 50 to 200 nm (data not shown). This is shown in
FIG. 13 .
Claims (16)
1-63. (canceled)
64. A hyaluronic acid derivative comprising n repeating units of formula (I):
wherein
(a) in at least one repeating unit, one or more of the R1, R2, R3, and R4 groups is an alkyl-succinic acid of formula (II):
65. The hyaluronic acid derivative of claim 64 , wherein one of the R1, R2, R3, and R4 groups is an alkyl-succinic acid of formula (II).
66. The hyaluronic acid derivative of claim 64 , wherein two of the R1, R2, R3, and R4 groups are an alkyl-succinic acid of formula (II).
67. The hyaluronic acid derivative of claim 64 , wherein three of the R1, R2, R3, and R4 groups are an alkyl-succinic acid of formula (II).
68. The hyaluronic acid derivative of claim 64 , wherein the alkyl group is C1-C20 alkyl group.
69. The hyaluronic acid derivative of claim 64 , wherein the alkyl group is propyl, 2-octenyl, 2-dodecenyl, 2-hexadecenyl, or 2-octadecenyl.
70. The hyaluronic acid derivative of claim 64 , which has an average molecular weight of between 10,000 and 1,500,000 Da.
71. The hyaluronic acid derivative of claim 64 , which has an average molecular weight of between 10,000 and 50,000 Da.
72. The hyaluronic acid derivative of claim 64 , which has an average molecular weight of between 50,000 and 500,000 Da.
73. A composition comprising a hyaluronic acid derivative of claim 64 and a water-soluble excipient.
74. The composition of claim 73 , wherein the excipient is lactose.
75. A pharmaceutical composition comprising an effective amount of a hyaluronic acid derivative of claim 64 and a pharmaceutically acceptable carrier, excipient or diluent.
76. A cosmetic article comprising a hyaluronic acid derivative of claim 64 .
77. A sanitary, medical or surgical article comprising a hyaluronic acid derivative of claim 64 , which is a diaper, a sanitary towel, a surgical sponge, a wound healing sponge, or a part comprised in a band aid or other wound dressing material.
78. A medicament capsule, microcapsule, nanocapsules, microsphere or nanosphere comprising a hyaluronic acid derivative of claim 64 .
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/164,012 US20110319357A1 (en) | 2005-09-26 | 2011-06-20 | Hyaluronic Acid Derivatives |
US13/554,190 US20120283213A1 (en) | 2005-09-26 | 2012-07-20 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
US14/017,085 US20140106411A1 (en) | 2005-09-26 | 2013-09-03 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DKPA200501332 | 2005-09-26 | ||
DKPA200501332 | 2005-09-26 | ||
US72123205P | 2005-09-27 | 2005-09-27 | |
PCT/DK2006/000523 WO2007033677A1 (en) | 2005-09-26 | 2006-09-26 | Aryl/alkyl succinic anhydride hyaluronan derivatives |
US57295407A | 2007-01-30 | 2007-01-30 | |
US13/164,012 US20110319357A1 (en) | 2005-09-26 | 2011-06-20 | Hyaluronic Acid Derivatives |
Related Parent Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2006/000523 Division WO2006078477A1 (en) | 2005-01-21 | 2006-01-06 | Air bag with gas blocking sealant at sewn seams |
US11/572,954 Division US7993678B2 (en) | 2005-09-26 | 2006-08-26 | Hyaluronic acid derivatives |
PCT/DK2006/000523 Division WO2007033677A1 (en) | 2005-09-26 | 2006-09-26 | Aryl/alkyl succinic anhydride hyaluronan derivatives |
US57295407A Division | 2005-09-26 | 2007-01-30 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/554,190 Continuation US20120283213A1 (en) | 2005-09-26 | 2012-07-20 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
Publications (1)
Publication Number | Publication Date |
---|---|
US20110319357A1 true US20110319357A1 (en) | 2011-12-29 |
Family
ID=37497045
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/572,954 Expired - Fee Related US7993678B2 (en) | 2005-09-26 | 2006-08-26 | Hyaluronic acid derivatives |
US13/164,012 Abandoned US20110319357A1 (en) | 2005-09-26 | 2011-06-20 | Hyaluronic Acid Derivatives |
US13/554,190 Abandoned US20120283213A1 (en) | 2005-09-26 | 2012-07-20 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
US14/017,085 Abandoned US20140106411A1 (en) | 2005-09-26 | 2013-09-03 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/572,954 Expired - Fee Related US7993678B2 (en) | 2005-09-26 | 2006-08-26 | Hyaluronic acid derivatives |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/554,190 Abandoned US20120283213A1 (en) | 2005-09-26 | 2012-07-20 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
US14/017,085 Abandoned US20140106411A1 (en) | 2005-09-26 | 2013-09-03 | Aryl/Alkyl Succinic Anhydride Hyaluronan Derivatives |
Country Status (9)
Country | Link |
---|---|
US (4) | US7993678B2 (en) |
EP (1) | EP1931713B1 (en) |
JP (1) | JP5155866B2 (en) |
CN (1) | CN101273065B (en) |
AU (1) | AU2006294207B2 (en) |
BR (1) | BRPI0616298A2 (en) |
CA (1) | CA2621899C (en) |
DK (1) | DK1931713T3 (en) |
WO (1) | WO2007033677A1 (en) |
Families Citing this family (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101594552B1 (en) | 2008-04-04 | 2016-02-17 | 유니버시티 오브 유타 리서치 파운데이션 | Alkylated sem-synthetic glycosaminoglycosan ethers, and methods for making and using thereof |
CZ2009835A3 (en) | 2009-12-11 | 2011-06-22 | Contipro C A.S. | Process for preparing hyaluronic acid derivative oxidized in position 6 of saccharide glucosamine portion selectively to aldehyde and modification method thereof |
CZ302504B6 (en) | 2009-12-11 | 2011-06-22 | Contipro C A.S. | Hyaluronic acid derivative oxidized selectively in position 6 of polysaccharide glucosamine portion to aldehyde and modification process thereof |
EP2688402B1 (en) | 2011-03-23 | 2018-10-24 | University of Utah Research Foundation | Means for treating or preventing urological inflammation |
KR20140034853A (en) | 2011-05-30 | 2014-03-20 | 노보자임스 바이오파마 디케이 에이/에스 | Spray drying of high molecular weight hyaluronic acid |
TWI561535B (en) | 2011-10-06 | 2016-12-11 | Bvw Holding Ag | Copolymers of hydrophobic and hydrophilic segments that reduce protein adsorption |
CZ303879B6 (en) | 2012-02-28 | 2013-06-05 | Contipro Biotech S.R.O. | Derivatives based on hyaluronic acid capable of forming hydrogels, process of their preparation, hydrogels based on these derivatives, process of their preparation and use |
CZ304512B6 (en) | 2012-08-08 | 2014-06-11 | Contipro Biotech S.R.O. | Hyaluronic acid derivative, process for its preparation, modification process and use thereof |
CZ2012842A3 (en) * | 2012-11-27 | 2014-08-20 | Contipro Biotech S.R.O. | C6-C18-acylated hyaluronate-based nanomicellar composition, process for preparing C6-C18-acylated hyaluronate, process for preparing nanomicellar composition and stabilized nanomicellar composition as well as use thereof |
US11439594B2 (en) | 2012-12-04 | 2022-09-13 | Phosphorex, Inc. | Microparticles and nanoparticles having negative surface charges |
US9879124B2 (en) * | 2013-08-29 | 2018-01-30 | Dainichiseika Color & Chemicals Mfg. Co., Ltd. | Method for manufacturing water-insoluble molded article and water-insoluble molded article |
ES2653549T3 (en) | 2013-09-30 | 2018-02-07 | Galderma S.A. | Functionalization and cross-linking of hyaluronic acid in one step |
US20150119505A1 (en) * | 2013-10-29 | 2015-04-30 | Edward Scott Williams | Paper Coating Composition |
CZ2014150A3 (en) | 2014-03-11 | 2015-05-20 | Contipro Biotech S.R.O. | Conjugates of hyaluronic acid oligomer or salts thereof, process of their preparation and use |
CZ2014451A3 (en) | 2014-06-30 | 2016-01-13 | Contipro Pharma A.S. | Antitumor composition based on hyaluronic acid and inorganic nanoparticles, process of its preparation and use |
WO2016136885A1 (en) * | 2015-02-27 | 2016-09-01 | 大日精化工業株式会社 | Method for manufacturing medical material, medical material, and anti-adhesion material |
CZ309295B6 (en) | 2015-03-09 | 2022-08-10 | Contipro A.S. | Self-supporting, biodegradable film based on hydrophobized hyaluronic acid, method of its preparation and use |
CZ306479B6 (en) | 2015-06-15 | 2017-02-08 | Contipro A.S. | A method of crosslinking polysaccharides by using photolabile protecting groups |
CZ306662B6 (en) | 2015-06-26 | 2017-04-26 | Contipro A.S. | Sulphated polysaccharides derivatives, the method of their preparation, the method of their modification and the use |
CZ308106B6 (en) | 2016-06-27 | 2020-01-08 | Contipro A.S. | Unsaturated derivatives of polysaccharides, preparing and using them |
GB201611695D0 (en) * | 2016-07-05 | 2016-08-17 | Statoil Petroleum As | Subsea wellhead installation and/or removal |
US11337994B2 (en) | 2016-09-15 | 2022-05-24 | University Of Utah Research Foundation | In situ gelling compositions for the treatment or prevention of inflammation and tissue damage |
CN108410926B (en) * | 2017-12-03 | 2021-12-14 | 新疆阜丰生物科技有限公司 | Method for preparing and extracting high molecular weight hyaluronic acid |
AR115689A1 (en) | 2018-07-05 | 2021-02-17 | Sumitomo Chemical Co | URACIL COMPOUNDS AND THEIR USE |
FR3083448B1 (en) * | 2018-07-06 | 2020-07-10 | Coatex | CLEANSING COSMETIC FORMULATION |
CN113750226B (en) * | 2021-08-12 | 2022-10-21 | 通用生物(安徽)股份有限公司 | Cationic lipid nucleic acid vaccine composition and preparation method thereof |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5356883A (en) * | 1989-08-01 | 1994-10-18 | Research Foundation Of State University Of N.Y. | Water-insoluble derivatives of hyaluronic acid and their methods of preparation and use |
US5416071A (en) * | 1991-03-12 | 1995-05-16 | Takeda Chemical Industries, Ltd. | Water-soluble composition for sustained-release containing epo and hyaluronic acid |
US6017901A (en) * | 1995-05-10 | 2000-01-25 | Fidia Advanced Bioplymers S.R.L. | Heavy metal salts of succinic acid hemiesters with hyaluronic acid or hyaluronic acid esters, a process for their preparation and relative pharmaceutical compositions |
US20080171083A1 (en) * | 2004-04-14 | 2008-07-17 | Vecture Limited | Pharmaceutical Compositions Comprising an Amphiphilic Starch |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5580553A (en) * | 1992-08-21 | 1996-12-03 | Nippon Starch Chemical Co., Ltd. | Cosmetic composition containing alkenylsuccinic acid ester of saccharide |
KR100279033B1 (en) | 1995-04-10 | 2001-03-02 | 존 맥도날드 | Polyglucosamine Derivatives, Preparation Method thereof, and Composition Comprising the Same |
IT1281877B1 (en) * | 1995-05-10 | 1998-03-03 | Fidia Advanced Biopolymers Srl | Heavy metal salts of succinyl derivatives of hyaluronic acid and their use as potential therapeutic agents |
HU226962B1 (en) * | 1995-08-29 | 2010-03-29 | Fidia Advanced Biopolymers Srl | Biomaterials for preventing post-surgical adhesions comprised of hyaluronic acid derivatives |
IT1286510B1 (en) * | 1996-11-29 | 1998-07-15 | Cooperativa Centro Ricerche Po | BUTYRIC ESTERS WITH ANTI-PROLIFERATIVE ACTIVITY AND PHARMACEUTICAL COMPOSITIONS THAT CONTAIN THEM |
US5977348A (en) | 1997-07-25 | 1999-11-02 | National Starch And Chemical Investment Holding Corporation | Polysaccharide modification in densified fluid |
US20020086852A1 (en) * | 1998-05-14 | 2002-07-04 | Cantor Jerome O. | Method for treating respiratory disorders associated with pulmonary elastic fiber injury |
GB9902412D0 (en) * | 1999-02-03 | 1999-03-24 | Fermentech Med Ltd | Process |
CN1136012C (en) * | 2000-08-07 | 2004-01-28 | 黄玲惠 | Wound dressing and its prepn. |
-
2006
- 2006-08-26 US US11/572,954 patent/US7993678B2/en not_active Expired - Fee Related
- 2006-09-26 AU AU2006294207A patent/AU2006294207B2/en not_active Ceased
- 2006-09-26 WO PCT/DK2006/000523 patent/WO2007033677A1/en active Application Filing
- 2006-09-26 EP EP06791421A patent/EP1931713B1/en not_active Not-in-force
- 2006-09-26 CA CA2621899A patent/CA2621899C/en not_active Expired - Fee Related
- 2006-09-26 BR BRPI0616298-3A patent/BRPI0616298A2/en not_active IP Right Cessation
- 2006-09-26 JP JP2008531534A patent/JP5155866B2/en not_active Expired - Fee Related
- 2006-09-26 CN CN2006800354457A patent/CN101273065B/en not_active Expired - Fee Related
- 2006-09-26 DK DK06791421.8T patent/DK1931713T3/en active
-
2011
- 2011-06-20 US US13/164,012 patent/US20110319357A1/en not_active Abandoned
-
2012
- 2012-07-20 US US13/554,190 patent/US20120283213A1/en not_active Abandoned
-
2013
- 2013-09-03 US US14/017,085 patent/US20140106411A1/en not_active Abandoned
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5356883A (en) * | 1989-08-01 | 1994-10-18 | Research Foundation Of State University Of N.Y. | Water-insoluble derivatives of hyaluronic acid and their methods of preparation and use |
US5416071A (en) * | 1991-03-12 | 1995-05-16 | Takeda Chemical Industries, Ltd. | Water-soluble composition for sustained-release containing epo and hyaluronic acid |
US6017901A (en) * | 1995-05-10 | 2000-01-25 | Fidia Advanced Bioplymers S.R.L. | Heavy metal salts of succinic acid hemiesters with hyaluronic acid or hyaluronic acid esters, a process for their preparation and relative pharmaceutical compositions |
US20080171083A1 (en) * | 2004-04-14 | 2008-07-17 | Vecture Limited | Pharmaceutical Compositions Comprising an Amphiphilic Starch |
Non-Patent Citations (1)
Title |
---|
Kawaguchi, Carbohydrate POlymers, 26, 1995 * |
Also Published As
Publication number | Publication date |
---|---|
JP2009510186A (en) | 2009-03-12 |
EP1931713B1 (en) | 2012-12-05 |
BRPI0616298A2 (en) | 2011-06-14 |
AU2006294207A1 (en) | 2007-03-29 |
WO2007033677A1 (en) | 2007-03-29 |
JP5155866B2 (en) | 2013-03-06 |
AU2006294207B2 (en) | 2012-11-29 |
CN101273065B (en) | 2011-12-28 |
CA2621899C (en) | 2014-05-06 |
US20090252810A1 (en) | 2009-10-08 |
EP1931713A1 (en) | 2008-06-18 |
US20120283213A1 (en) | 2012-11-08 |
DK1931713T3 (en) | 2013-03-11 |
US20140106411A1 (en) | 2014-04-17 |
CA2621899A1 (en) | 2007-03-29 |
US7993678B2 (en) | 2011-08-09 |
CN101273065A (en) | 2008-09-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7993678B2 (en) | Hyaluronic acid derivatives | |
EP1994062B1 (en) | Aryl/alkyl vinyl sulfone hyaluronic acid derivatives | |
US20100210588A1 (en) | Hyaluronic Acid Linked with a Polymer of an Alpha Hydroxy Acid | |
JP5313497B2 (en) | Dried and agglomerated hyaluronic acid product | |
US7956180B2 (en) | Dried and agglomerated hyaluronic acid product | |
US20050272695A1 (en) | Fast dissolving dried hyaluronic acid product | |
WO2005116130A1 (en) | A fast dissolving dried hyaluronic acid product | |
KR20070017374A (en) | A fast dissolving dried hyaluronic acid product |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |