HRP930063A2 - Novel compositions containing hyaluronic acid associates and a process for preparing the same - Google Patents
Novel compositions containing hyaluronic acid associates and a process for preparing the same Download PDFInfo
- Publication number
- HRP930063A2 HRP930063A2 HR930063A HRP930063A HRP930063A2 HR P930063 A2 HRP930063 A2 HR P930063A2 HR 930063 A HR930063 A HR 930063A HR P930063 A HRP930063 A HR P930063A HR P930063 A2 HRP930063 A2 HR P930063A2
- Authority
- HR
- Croatia
- Prior art keywords
- zinc
- hyaluronate
- complex
- hyaluronic acid
- solution
- Prior art date
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 compound 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 description 39
- 229920002674 hyaluronan Polymers 0.000 title claims description 26
- 229960003160 hyaluronic acid Drugs 0.000 title claims description 26
- 239000000203 mixture Substances 0.000 title claims description 5
- 238000004519 manufacturing process Methods 0.000 title claims 2
- 239000000243 solution Substances 0.000 claims description 98
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 74
- 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 claims description 66
- 229920002385 Sodium hyaluronate Polymers 0.000 claims description 44
- 229940010747 sodium hyaluronate Drugs 0.000 claims description 44
- 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 claims description 44
- 238000002360 preparation method Methods 0.000 claims description 43
- 239000011592 zinc chloride Substances 0.000 claims description 37
- 235000005074 zinc chloride Nutrition 0.000 claims description 37
- 238000000034 method Methods 0.000 claims description 34
- 229910021645 metal ion Inorganic materials 0.000 claims description 32
- 239000007864 aqueous solution Substances 0.000 claims description 31
- 239000004480 active ingredient Substances 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 229940014041 hyaluronate Drugs 0.000 claims description 27
- 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 claims description 26
- 230000000737 periodic effect Effects 0.000 claims description 23
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims description 17
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 14
- 229910052725 zinc Inorganic materials 0.000 claims description 14
- 239000011701 zinc Substances 0.000 claims description 13
- 230000008569 process Effects 0.000 claims description 12
- 239000002537 cosmetic Substances 0.000 claims description 11
- 150000003839 salts Chemical class 0.000 claims description 11
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 239000000654 additive Substances 0.000 claims description 5
- 229910052784 alkaline earth metal Inorganic materials 0.000 claims description 5
- 230000001225 therapeutic effect Effects 0.000 claims description 5
- 239000000969 carrier Substances 0.000 claims description 4
- 229910001429 cobalt ion Inorganic materials 0.000 claims description 4
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 claims description 4
- 150000003242 quaternary ammonium salts Chemical class 0.000 claims description 4
- GGCZERPQGJTIQP-UHFFFAOYSA-N sodium;9,10-dioxoanthracene-2-sulfonic acid Chemical compound [Na+].C1=CC=C2C(=O)C3=CC(S(=O)(=O)O)=CC=C3C(=O)C2=C1 GGCZERPQGJTIQP-UHFFFAOYSA-N 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 150000001342 alkaline earth metals Chemical class 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 2
- 239000007900 aqueous suspension Substances 0.000 claims description 2
- 150000004700 cobalt complex Chemical class 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims 4
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 238000002844 melting Methods 0.000 claims 1
- 230000008018 melting Effects 0.000 claims 1
- 239000012153 distilled water Substances 0.000 description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 20
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 18
- 229910001415 sodium ion Inorganic materials 0.000 description 15
- 238000011282 treatment Methods 0.000 description 15
- 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 12
- 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 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 208000025865 Ulcer Diseases 0.000 description 12
- 229960002920 sorbitol Drugs 0.000 description 12
- 231100000397 ulcer Toxicity 0.000 description 12
- 239000000600 sorbitol Substances 0.000 description 11
- 239000011780 sodium chloride Substances 0.000 description 10
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 150000002500 ions Chemical class 0.000 description 9
- 238000005259 measurement Methods 0.000 description 9
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 8
- 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 8
- 239000008103 glucose Substances 0.000 description 8
- 230000003204 osmotic effect Effects 0.000 description 8
- 239000004302 potassium sorbate Substances 0.000 description 8
- 235000010241 potassium sorbate Nutrition 0.000 description 8
- 229940069338 potassium sorbate Drugs 0.000 description 8
- 150000001875 compounds Chemical class 0.000 description 7
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- -1 ammonium ions Chemical class 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 239000011148 porous material Substances 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 206010052428 Wound Diseases 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 229910017052 cobalt Inorganic materials 0.000 description 4
- 239000010941 cobalt Substances 0.000 description 4
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 description 4
- GLDOVTGHNKAZLK-UHFFFAOYSA-N octadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCO GLDOVTGHNKAZLK-UHFFFAOYSA-N 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 3
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 239000001632 sodium acetate Substances 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 159000000000 sodium salts Chemical class 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 150000005846 sugar alcohols Chemical class 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 210000000689 upper leg Anatomy 0.000 description 3
- 239000003871 white petrolatum Substances 0.000 description 3
- FLPJVCMIKUWSDR-UHFFFAOYSA-N 2-(4-formylphenoxy)acetamide Chemical compound NC(=O)COC1=CC=C(C=O)C=C1 FLPJVCMIKUWSDR-UHFFFAOYSA-N 0.000 description 2
- QCDWFXQBSFUVSP-UHFFFAOYSA-N 2-phenoxyethanol Chemical compound OCCOC1=CC=CC=C1 QCDWFXQBSFUVSP-UHFFFAOYSA-N 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000167854 Bourreria succulenta Species 0.000 description 2
- DCXXMTOCNZCJGO-UHFFFAOYSA-N Glycerol trioctadecanoate Natural products CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 229920002125 Sokalan® Polymers 0.000 description 2
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 206010000496 acne Diseases 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 239000007765 cera alba Substances 0.000 description 2
- 229960000541 cetyl alcohol Drugs 0.000 description 2
- 229940074979 cetyl palmitate Drugs 0.000 description 2
- YMKDRGPMQRFJGP-UHFFFAOYSA-M cetylpyridinium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 YMKDRGPMQRFJGP-UHFFFAOYSA-M 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 235000019693 cherries Nutrition 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 229940075507 glyceryl monostearate Drugs 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- PXDJXZJSCPSGGI-UHFFFAOYSA-N hexadecanoic acid hexadecyl ester Natural products CCCCCCCCCCCCCCCCOC(=O)CCCCCCCCCCCCCCC PXDJXZJSCPSGGI-UHFFFAOYSA-N 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 2
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 2
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 2
- 239000001788 mono and diglycerides of fatty acids Substances 0.000 description 2
- GOQYKNQRPGWPLP-UHFFFAOYSA-N n-heptadecyl alcohol Natural products CCCCCCCCCCCCCCCCCO GOQYKNQRPGWPLP-UHFFFAOYSA-N 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 229960005323 phenoxyethanol Drugs 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 2
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 2
- 229950004959 sorbitan oleate Drugs 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OULAJFUGPPVRBK-UHFFFAOYSA-N tetratriacontan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCO OULAJFUGPPVRBK-UHFFFAOYSA-N 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- MPNXSZJPSVBLHP-UHFFFAOYSA-N 2-chloro-n-phenylpyridine-3-carboxamide Chemical compound ClC1=NC=CC=C1C(=O)NC1=CC=CC=C1 MPNXSZJPSVBLHP-UHFFFAOYSA-N 0.000 description 1
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical group [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 229910002480 Cu-O Inorganic materials 0.000 description 1
- 206010011985 Decubitus ulcer Diseases 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 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
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000004166 Lanolin Substances 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
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- 229910007541 Zn O Inorganic materials 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910052768 actinide Inorganic materials 0.000 description 1
- 150000001255 actinides Chemical class 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 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 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
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- 206010003246 arthritis Diseases 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- UREZNYTWGJKWBI-UHFFFAOYSA-M benzethonium chloride Chemical compound [Cl-].C1=CC(C(C)(C)CC(C)(C)C)=CC=C1OCCOCC[N+](C)(C)CC1=CC=CC=C1 UREZNYTWGJKWBI-UHFFFAOYSA-M 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000007942 carboxylates Chemical group 0.000 description 1
- 229960001927 cetylpyridinium chloride Drugs 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- SXYCCJAPZKHOLS-UHFFFAOYSA-N chembl2008674 Chemical compound [O-][N+](=O)C1=CC=C2C(N=NC3=C4C=CC=CC4=CC=C3O)=C(O)C=C(S(O)(=O)=O)C2=C1 SXYCCJAPZKHOLS-UHFFFAOYSA-N 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
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- 230000000052 comparative effect Effects 0.000 description 1
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- 125000005448 ethoxyethyl group Chemical group [H]C([H])([H])C([H])([H])OC([H])([H])C([H])([H])* 0.000 description 1
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- MSWZFWKMSRAUBD-IVMDWMLBSA-N glucosamine group Chemical group OC1[C@H](N)[C@@H](O)[C@H](O)[C@H](O1)CO MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
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- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 235000019388 lanolin Nutrition 0.000 description 1
- 229940039717 lanolin Drugs 0.000 description 1
- 229910052747 lanthanoid Inorganic materials 0.000 description 1
- 150000002602 lanthanoids Chemical class 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
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- 235000019808 microcrystalline wax Nutrition 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
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- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
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- 102000004169 proteins and genes Human genes 0.000 description 1
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- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 150000003752 zinc compounds Chemical class 0.000 description 1
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- Polysaccharides And Polysaccharide Derivatives (AREA)
Description
Izum se odnosi na nove veze (komplekse) deprotonizirane hijaluronske kiseline sa 3d metalnim ionima četvrte periode periodnog sustava i na spojeve, koji sadrže te komplekse kao aktivne sastojke. The invention relates to new bonds (complexes) of deprotonated hyaluronic acid with 3d metal ions of the fourth period of the periodic table and to compounds containing these complexes as active ingredients.
Nadalje, izum se odnosi na postupak za pripravu tih novih asocijacija (kompleksa) i na spojeve, koji te komplekse sadrže kao aktivne sastojke. Furthermore, the invention relates to the process for the preparation of these new associations (complexes) and to compounds, which contain these complexes as active ingredients.
Prema vrlo osebujnoj izvedbi postupka predloženog izuma vodene otopine, koje sadrže nove komplekse deprotonizirane hijaluronske kiseline sa 3d metalnim ionima četvrte periode periodnog sustava, pripravljamo neposredno iz vodenih otopina natrijevog hijaluronata. According to a very special embodiment of the procedure of the proposed invention, aqueous solutions containing new complexes of deprotonated hyaluronic acid with 3d metal ions of the fourth period of the periodic table are prepared directly from aqueous solutions of sodium hyaluronate.
Novi kompleksi prema predloženom izumu uključuju uglavnom cinkov i kobaltov hijaluronat. Spojevi, koji sadrže te zadnje komplekse, mogu biti farmaceutski (terapeutski) ili kozmetički i u danom primjeru drugi spojevi. Područja indikacije, koja sadrže spojeve novih kompleksa u smislu izuma, su npr.: ubrzavanje epitelizacije na površini tijela gdje nedostaje epitel, liječenje ulkusa na bedru, dekubitusa, osobito nezaraslih rana, opekotina, zračenjem ili temperaturom uzrokovanih rana, uobičajenih akna i sistemskih akna, kao i iz drugih područja. The new complexes according to the proposed invention include mainly zinc and cobalt hyaluronate. The compounds, which contain these last complexes, can be pharmaceutical (therapeutic) or cosmetic and in the given example other compounds. Areas of indication, which contain compounds of new complexes in terms of the invention, are, for example: acceleration of epithelization on the body surface where epithelium is missing, treatment of thigh ulcers, decubitus, especially unhealed wounds, burns, wounds caused by radiation or temperature, common acne and systemic acne, as well as from other areas.
Hijaluronska kiselina je makromolekula, koja je poznata već više od 50 godina, a prvi su je opisali Mayer et al. (J. Biol. Chem. 107, 629 (1954): J. Biol. Chem. 114, 689 (1936). Strukturu je odredio Weissman et al. (J. Am. Chem. Soc. 76, 1753 (1954)). Hijaluronska je kiselina prirodni glukozamino glikan sa visokim viskozitetom, koji sadrži izmjenično β1-3 glukuronsku kiselinu i β1-4 glukozaminske ostatke: njena molekularna masa je od 50000 do više milijuna (8 do 13 milijuna). Dobivanje hijaluronske kiseline je staro područje. Odvajanje i uporaba posebno čiste hijaluronske kiseline su opisani npr. u patetnim spisima Ujedinjenih Država Amerike broj 4,141,973 i 4 303 676 i u europskom patentnom spisu broj 0 144 019. Do pred nekoliko godina hijaluronsku kiselinu su koristili kao natrijevu sol, npr. terapijski uglavnom u oftalmologiji, kirurgiji i kozmetici. Soli hijaluronske kiseline nastale sa ionima alkalijskih, zemnoalkalijskih metala, magnezija, aluminija, amonijaka ili supstituiranih amonijevih iona, mogu se koristiti kao nosioci za povećanje apsorpcije lijekova (vidi belgijski patentni spis broj 904,547). Soli teških metala hijaluronske kiseline (gdje "teški metali" znače elemente pete, šeste i sedme periode periodnog sustava kao i lantanide te aktinide) i među njima srebrnu sol, upotrebljavali su kao fungicidno sredstvo, dok su srebrnu sol koristili u liječenju artritisa (vidi patentni spis WO 87/05517). Hyaluronic acid is a macromolecule, which has been known for more than 50 years, and was first described by Mayer et al. (J. Biol. Chem. 107, 629 (1954): J. Biol. Chem. 114, 689 (1936). The structure was determined by Weissman et al. (J. Am. Chem. Soc. 76, 1753 (1954)) . Hyaluronic acid is a natural glucosaminoglycan with high viscosity, containing alternating β1-3 glucuronic acid and β1-4 glucosamine residues: its molecular weight is from 50,000 to several million (8 to 13 million). Obtaining hyaluronic acid is an old field. Separation and the use of particularly pure hyaluronic acid are described, for example, in the United States Patent No. 4,141,973 and 4,303,676 and in the European Patent No. 0,144,019. Until a few years ago, hyaluronic acid was used as a sodium salt, e.g. therapeutically mainly in ophthalmology, surgery and cosmetics. Salts of hyaluronic acid formed with ions of alkali, alkaline earth metals, magnesium, aluminum, ammonia or substituted ammonium ions can be used as carriers to increase the absorption of drugs (see Belgian patent document bro j 904,547). Heavy metal salts of hyaluronic acid (where "heavy metals" mean the elements of the fifth, sixth and seventh periods of the periodic table as well as lanthanides and actinides) and among them silver salt, were used as a fungicidal agent, while silver salt was used in the treatment of arthritis (see patent file WO 87/05517).
Različitim metodama određivanja strukture bilo je dokazano, da se sekundarna struktura, t.j. konformacija hijaluronske kiseline, promijeni pri različitim metodama određivanja strukture bilo je dokazano, da se sekundarna struktura, t.j. konformacija hijaluronske kiseline, promijeni pri vezanju metalnih iona (W. T. Winter i A. Sruther: J. Mol. Biol. 517, 761 (1977); J. K. Sheehan i E. D. T. Atkins: Int. J. Biol. Macromol. 5, 215 (1983); i N. Figueroa i B. Chakrabarti: Bioplymers 17, 2415 (1978). Signifikantnu promjenu učinaka na molekularnu strukturu mogu izazvati i metalni ioni sličnog značaja, kako se pokazalo usporednom X-ray studijom kalijevog i natrijevog hijaluronata (A. K. Mitra et al,: J. Macromol., Sci. Phys. 824, 1 i 21 (1985). To još više vrijedi za spojeve hijaluronske kiseline nastale sa metalnim ionima različitih vrsta, koji nose različite naboje. Various methods of determining the structure proved that the secondary structure, i.e. the conformation of hyaluronic acid changes during different methods of determining the structure, it was proved that the secondary structure, i.e. conformation of hyaluronic acid changes upon metal ion binding (W. T. Winter and A. Sruther: J. Mol. Biol. 517, 761 (1977); J. K. Sheehan and E. D. T. Atkins: Int. J. Biol. Macromol. 5, 215 (1983) ; and N. Figueroa and B. Chakrabarti: Bioplymers 17, 2415 (1978). A significant change in the effects on the molecular structure can also be caused by metal ions of similar importance, as shown by a comparative X-ray study of potassium and sodium hyaluronate (A. K. Mitra et al, : J. Macromol., Sci. Phys. 824, 1 and 21 (1985). This is even more true for compounds of hyaluronic acid formed with metal ions of different types, which carry different charges.
U literaturi nismo našli nikakve reference, koje bi se odnosile na veze (komplekse) hijaluronske kiseline nastale sa 3d metalnim ionima četvrte periode periodnog sustava; glede istraživanja gelske filtracijske kromatografije hijaluronske kiseline, suprotno heparinu, ona ne veže cinkove ione (R. F. Parish i W. R. Fair: Biochem J. 193, 407-410 (1981). We did not find any references in the literature, which would refer to the connections (complexes) of hyaluronic acid formed with 3d metal ions of the fourth period of the periodic table; in gel filtration chromatography studies of hyaluronic acid, unlike heparin, it does not bind zinc ions (R.F. Parish and W.R. Fair: Biochem J. 193, 407-410 (1981).
Unatoč činjenici da, s obzirom na literaturu, hijaluronska kiselina (ili njena natrijeva sol) ne veže cinkove ione, autori su predloženog izuma ispitali koordinacijsku kemiju interakcije između hijaluronske kiseline i 3d metalnih iona četvrte periode periodnog sustava a unutar njih poglavito ione cinka i kobalta. Kako se hijaluronska kiselina komercijalno nabavlja skoro isključivo kao njena natrijeva sol, a kao takova je osnovna supstanca za sve sustave koji sadrže hijaluronat, naša smo istraživanja počeli pri interakciji natrijevih iona i hijaluronata. U tu svrhu smo mjerili aktivnost slobodnog natrijevog iona vodenih otopina natrijevog hijaluronata sa uporabom natrijeve selektivne staklene elektrode. Tm mjerenjem smo nedvosmisleno ustanovili da nije prisutno više od 60% uvedenih natrijevih iona kao ekvivalent sa karboksilantnim skupinama hijaluronata u obliku slobodnih iona u vodenim otopinama, dok je ostalih 40% u obliku vezanih za hijaluronat. Obzirom na naša mjerenja, sa rastom koncentracije natrijevog iona mnogo vezanih natrijevih iona možemo podići na 50-55 %, izračunato za sve raspoložive karboksilatne skupine. Tako smo potvrdili da, za razliku od zajedničkih osobina soli, natrijev hijaluronat u vodenim otopinama nije potpuno Despite the fact that, according to the literature, hyaluronic acid (or its sodium salt) does not bind zinc ions, the authors of the proposed invention examined the coordination chemistry of the interaction between hyaluronic acid and 3d metal ions of the fourth period of the periodic table, and within them mainly zinc and cobalt ions. Since hyaluronic acid is commercially available almost exclusively as its sodium salt, and as such is the basic substance for all systems containing hyaluronate, we started our research on the interaction of sodium ions and hyaluronate. For this purpose, we measured the free sodium ion activity of sodium hyaluronate aqueous solutions using a sodium selective glass electrode. Through this measurement, we unequivocally established that no more than 60% of the introduced sodium ions are present as equivalent to the carboxyl groups of hyaluronate in the form of free ions in aqueous solutions, while the other 40% are in the form bound to hyaluronate. Considering our measurements, with the growth of the sodium ion concentration, we can raise the number of bound sodium ions to 50-55%, calculated for all available carboxylate groups. Thus, we confirmed that, in contrast to the common properties of salt, sodium hyaluronate in aqueous solutions is not complete
disociran. dissociated.
U slijedećem stupnju naših istraživanja vodenu smo otopinu natrijevog hijaluronata titrirali sa otopinom cinkovog klorida uporabom gore spomenute ion-selektivne elektrode za natrijeve ione, da bi pratili promjene aktivnosti slobodnih natrijevih iona u sustavu. Značajna krivulja, koja prikazuje taj postupak, prikazana je na slici 1. In the next stage of our research, we titrated an aqueous solution of sodium hyaluronate with a zinc chloride solution using the aforementioned ion-selective electrode for sodium ions, in order to monitor changes in the activity of free sodium ions in the system. A significant curve, which shows this process, is shown in Figure 1.
Opaža se da se natrijevi ioni, prethodno vezani na hijaluronat, oslobađaju zbog učinka cinkovih iona. Na osnovi rezultata tih mjerenja cjelokupnu smo koncentraciju natrijevih iona oslobodili sa ekvivalentnom količinom cinkovih, činjenica, koja nedvosmisleno dokazuje da su cinkovi ioni jače vezani na hijaluronat nego natrijevi ioni. Tako smo eksperimentalno opovrgli prijašnju tvrdnju, da hijaluronska kiselina ne može vezati cinkove ione (R. F. Parrish i W. R. Fair: Biochem. J. 193, 407 (1981)). It is observed that sodium ions, previously bound to hyaluronate, are released due to the effect of zinc ions. Based on the results of these measurements, we released the entire concentration of sodium ions with an equivalent amount of zinc, a fact that unequivocally proves that zinc ions are more strongly bound to hyaluronate than sodium ions. Thus, we experimentally refuted the previous claim that hyaluronic acid cannot bind zinc ions (R.F. Parrish and W.R. Fair: Biochem. J. 193, 407 (1981)).
Time smo opovrgli tvrdnju stručnjaka, koja je vrijedila do sada. With this, we refuted the expert's claim, which was valid until now.
Iz naših gore navedenih istraživanja postalo je vidljivo, da se pri interakciji ekvivalentnih količina natrijevog hijaluronata i cinkovih iona (cinkovog klorida) tvori u vodenoj otopini kompleks cinkovog hijaluronata sa stehiomeričnim sastavom. Po odgovarajućoj izotonizaciji možemo dobivenu otopinu direktno neposredno upotrijebiti u terapeutske namjene i cinkov spoj nije potrebno pripraviti u tvrdom stanju u odvojenom postupku. Prethodna istraživanja izvedena sa uporabom kobaltnog iona i drugih 3d metalnih iona dovele su do sličnih rezultata. Ipak smo kompleks pripravili u tvrdom stanju sa karakterizacijom i neposrednim određivanjem okoline cinkovog iona sa uporabom metode "Extended X-ray Absorption Fine Structure" (EXAFS). Možemo zaključiti, da je cinkov ion okružen sa četiri kisikova atoma u prvoj koordinacijskoj sferi. Dužina Zn-O veze je 1,99 pm, dok su dva ugljična atoma prisutna na većoj udaljenosti 241 pm od cinkovog atoma. From our research mentioned above, it became evident that when equivalent amounts of sodium hyaluronate and zinc ions (zinc chloride) interact, a complex of zinc hyaluronate with a stoichiometric composition is formed in an aqueous solution. After appropriate isotonization, the obtained solution can be directly used for therapeutic purposes and it is not necessary to prepare the zinc compound in a solid state in a separate procedure. Previous research conducted using cobalt ion and other 3d metal ions led to similar results. Nevertheless, we prepared the complex in the solid state with characterization and direct determination of the environment of the zinc ion using the "Extended X-ray Absorption Fine Structure" (EXAFS) method. We can conclude that the zinc ion is surrounded by four oxygen atoms in the first coordination sphere. The length of the Zn-O bond is 1.99 pm, while the two carbon atoms are present at a greater distance of 241 pm from the zinc atom.
Obzirom na naša istraživanja cinkov se hijaluronat bitno razlikuje od sličnih kompleksa bakra, koji sadrže četiri ekvatorijalne i dvije aksijalne Cu-O veze sa vrijednostima 1,94 i 234 pm. Udaljenost između atoma bakra i susjednih atoma ugljika je 258 pm. Struktura kobaltnog kompleksa je slična cinkovom kompleksu, ali ne i kompleksu bakra. Tako se predloženi izum odnosi na veze (komplekse) deprotonizirane hijaluronske kiseline sa 3d metalnim ionima četvrte periode periodnog sustava. Based on our research, zinc hyaluronate is significantly different from similar copper complexes, which contain four equatorial and two axial Cu-O bonds with values of 1.94 and 234 pm. The distance between copper atoms and neighboring carbon atoms is 258 pm. The structure of the cobalt complex is similar to the zinc complex, but not to the copper complex. Thus, the proposed invention relates to bonds (complexes) of deprotonated hyaluronic acid with 3d metal ions of the fourth period of the periodic table.
Izum se nadalje odnosi na sastav, koji sadrži aktivni sastojak ili nosilac kompleks hijaluronske kiseline sa 3d metalnim ionima četvrte periode periodnog sustava, u danom primjeru u primjesi sa drugim aktivnim sastojcima i/ili dodacima. Izum se isto tako odnosi na postupak za pripravljanje novih kompleksa u smislu izuma, koji obuhvaća The invention further relates to the composition, which contains the active ingredient or carrier complex of hyaluronic acid with 3d metal ions of the fourth period of the periodic table, in the given example in admixture with other active ingredients and/or additives. The invention also relates to a process for the preparation of new complexes within the meaning of the invention, which it encompasses
a) dodavanje vodene otopine, koja sadrži ekvivalentnu količinu soli ponajprije klorida, jednog od 3d metalnih iona četvrte periode periodnog sustava u vodenu otopinu natrijevog hijaluronata ili kakve druge soli (alkalijskog ili zemnoalkalijskog metala, u danom primjeru srebrne soli) hijaluronata; ili a) adding an aqueous solution, which contains an equivalent amount of salt, preferably chloride, one of the 3d metal ions of the fourth period of the periodic table to an aqueous solution of sodium hyaluronate or some other salt (alkali or alkaline earth metal, in this example silver salt) of hyaluronate; or
b) otapljanje kompleksa nastalog iz hijaluronske kiseline sa kvaternarnom amonijevom soli u vodenoj suspenziji u paru otapala, koji sadrži vodenu otopinu 3d metalnog iona četvrte periode periodnog sustava i otapala, koji se djelomično miješa sa vodom, ponajprije n-butanola; i zatim b) dissolving the complex formed from hyaluronic acid with a quaternary ammonium salt in an aqueous suspension in a pair of solvents, which contains an aqueous solution of the 3d metal ion of the fourth period of the periodic table and a solvent that is partially miscible with water, primarily n-butanol; and then
- taloženje kompleksa, dobivenog iz hijaluronske kiseline sa 3d metalnim ionom četvrte periode periodnog sustava sa alkanolom ili alkanonom na poznati način, ili - precipitation of the complex obtained from hyaluronic acid with a 3d metal ion of the fourth period of the periodic table with alkanol or alkanone in a known manner, or
- odvajanje taloga iz otopine i zatim, ukoliko želimo - separation of the precipitate from the solution and then, if desired
- sušenje pod blagim uvjetima. - drying under mild conditions.
Na osnovi gore navedenog razvili smo postupak za pripravljanje vodenih otopina, koje sadrže kao aktivno sredstvo kompleks cinkovog hijaluronata ili sličnog kompleksa 3d metalnih iona četvrte periode periodnog sustava. Te otopine pripravljamo svaki puta sa neposrednom reakcijom metalnih iona sa komponentom hijaluronata. Po toj metodi pripravljanja nije potrebno prethodno odvajanje aktivnih sastojaka u tvrdom stanju. U otopini pripravljenoj za uporabu postupka u smislu izuma je količina slobodnog (nevezanog sa metalom) hijaluronata zanemariva čak i u prisutnosti ekvivalentne količine cinka. U slučaju suviška cinkovih iona, nastanak je kompleksa cinkovog hijaluronata kvantitativan. On the basis of the above, we have developed a procedure for preparing aqueous solutions, which contain zinc hyaluronate complex or a similar complex of 3d metal ions of the fourth period of the periodic table as an active agent. We prepare these solutions every time with the direct reaction of metal ions with the hyaluronate component. According to this method of preparation, it is not necessary to separate the active ingredients in the solid state beforehand. In the solution prepared for the use of the method according to the invention, the amount of free (not bound to the metal) hyaluronate is negligible even in the presence of an equivalent amount of zinc. In the case of an excess of zinc ions, the formation of the zinc hyaluronate complex is quantitative.
U postupku pripravljanja metalnih kompleksa na gore opisan način ostaje pH-vrijednost oko 5. U slučaju 0,2 mas.%/vol.(ms./vol) dosegne pH vrijednost otopine hijaluronata vrijednost 5,4, dok je u slučaju 0,5 mas.%/vol. ta pH vrijednost 5. Ukoliko je potrebno možemo pH tog zadnjeg sustava poravnati na vrijednost od 5,5 do 5,6 sa dodatkom nekoliko kapljica izotonične otopine natrijevog acetata. In the process of preparing metal complexes as described above, the pH value remains around 5. In the case of 0.2 wt.%/vol. (ms./vol), the pH value of the hyaluronate solution reaches 5.4, while in the case of 0.5 wt.%/vol. that pH value is 5. If necessary, we can adjust the pH of the last system to a value of 5.5 to 5.6 with the addition of a few drops of isotonic sodium acetate solution.
Pripravili smo dvije vrste otopina, koje su sadržavale cinkov hijaluronat kao aktivni sastav sa uporabom gore opisanog postupka. We prepared two types of solutions, which contained zinc hyaluronate as an active ingredient, using the procedure described above.
1. Otopina cinkovog hijaluronata pripravljena izotonično sa viškom cinkovog klorida: 1. Zinc hyaluronate solution prepared isotonically with excess zinc chloride:
Ako znamo da možemo primarno upotrijebiti cinkov klorid u slobodnom obliku u dermatologiji, osmotski se tlak otopine cinkovog hijaluronata poravna na izotonične vrijednosti sa korištenjem suviška cinkovog klorida. If we know that we can primarily use zinc chloride in free form in dermatology, the osmotic pressure of the zinc hyaluronate solution is adjusted to isotonic values with the use of excess zinc chloride.
Tako dobivena otopina ne sadrži više ništa slobodnog na (cink-nevezanog) hijaluronata, nego je prisutan suvišak cinkovog klorida u sustavu skupa sa cinkovim hijaluronatom. The resulting solution no longer contains any free (zinc-unbound) hyaluronate, but an excess of zinc chloride is present in the system together with zinc hyaluronate.
2. Otopina cinkovog hijaluronata napravljena izotonično sa monosaharidom ili šećernim alkoholom: 2. Zinc hyaluronate solution made isotonic with monosaccharide or sugar alcohol:
Za terapeutsku upotrebu gdje nije označena prisutnost na hijaluronat-nevezanih cinkovih iona, napravimo otopinu, koja sadrži cinkove ione u ekvivalentnoj količini glede na hijaluronat izotoničnu sa upotrebom polialkohola (šećerni alkohol, ponajprije sorbitol) ili mono- ili di- saharida (ponajprije glukoze). Sadržaj slobodnog cinkovog iona i slobodnog hijaluronata u tim zadnjim sustavima ne doseže 5% cjelokupnog sadržaja cinka odnosno cjelokupnog sadržaja hijaluronata. For therapeutic use where the presence of hyaluronate-unbound zinc ions is not indicated, we make a solution containing zinc ions in an equivalent amount relative to hyaluronate isotonic with the use of polyalcohol (sugar alcohol, primarily sorbitol) or mono- or di-saccharides (primarily glucose). The content of free zinc ion and free hyaluronate in these latter systems does not reach 5% of the total zinc content, that is, of the total hyaluronate content.
U postupku uporabe kompleksa u smislu izuma mogu se zahtijevati i sastojci potpuno bez iona. Kompleksi, pripravljeni po gore navedenom postupku u smislu izuma naime uobičajeno sadrže natrijev klorid ili kakvu drugu sol nastalu iz ishodišnog hijaluronatnog kationa i aniona soli metala 3d. In the process of using the complex according to the invention, completely ion-free ingredients may be required. The complexes, prepared according to the above-mentioned process in terms of the invention, usually contain sodium chloride or some other salt formed from the initial hyaluronate cation and the anion of the metal salt 3d.
Možemo uporabiti dvije različite varijante postupka za pripravljanje kompleksa hijaluronske kiseline nastalih sa 3d metalnim ionima bez soli. Te su kako slijedi: We can use two different variants of the procedure for the preparation of hyaluronic acid complexes formed with 3d metal ions without salts. These are as follows:
a) Otopini kvaternarne amonijeve soli po dijelovima dodajemo otopinu poznatog hijaluronata, ponajprije natrijevog hijaluronata. Nakon zadovoljavajućeg čišćenja, novi istaloženi kompleks kvaternarnog amonijevog hijaluronata otopimo uz snažno miješanje u paru otapala, koji sastoji iz vodene otopine 3d metalnog iona četvrte periode periodnog sustava i otapala, koji se djelomično miješa sa vodom, ponajprije n-butanola. Ostavimo da se obje faze odvoje, nakon toga istaložimo kompleks hijaluronata sa dodatkom alkanola ili alkanona u vodenoj fazi, talog odvojimo i isperemo; ili a) To the quaternary ammonium salt solution, we add a solution of known hyaluronate, preferably sodium hyaluronate, in parts. After satisfactory cleaning, we dissolve the new precipitated complex of quaternary ammonium hyaluronate with vigorous stirring in solvent vapor, which consists of an aqueous solution of the 3d metal ion of the fourth period of the periodic table and a solvent that is partially miscible with water, primarily n-butanol. Let both phases separate, then precipitate the hyaluronate complex with the addition of alkanol or alkanone in the aqueous phase, separate the precipitate and wash it; or
b) po dodatku 2,0 do 3 volumena C1-3 alkanola ili C3-4 alkanona između miješanja u otopini cinkovog hijaluronata, prikladno u neizotoniziranoj otopini, koja sadrži cinkov klorid u količini ekvivalentnoj hijaluronatu, talog cinkovog hijaluronata filtriramo i isperemo sa alkanolom ili alkanonom, kojega smo upotrijebili za taloženje. Ukoliko je potrebno cinkov hijaluronat otopimo u vodi bez iona i taloženje ponovimo. b) by adding 2.0 to 3 volumes of C1-3 alkanol or C3-4 alkanone between mixing in a zinc hyaluronate solution, preferably in a non-isotonized solution, containing zinc chloride in an amount equivalent to hyaluronate, the zinc hyaluronate precipitate is filtered and washed with alkanol or alkanone , which we used for deposition. If necessary, dissolve zinc hyaluronate in water without ions and repeat the precipitation.
Ukoliko trebamo tvrdi cinkov hijaluronat bez iona, talog sušimo pod smanjenim tlakom i blagim uvjetima. U slučaju da se zahtijeva otopina cinkovog hijaluronata bez iona, bolje je da otopimo cinkov hijaluronat, koji ne sadrži otapala. Bilo kojom od obe varijante postupka dobijemo tvrdu tvar bez iona ili otopljen produkt sa čistoćom, koja u danom primjeru ovisi o kvaliteti ishodišnog cinkovog hijaluronata. If we need hard zinc hyaluronate without ions, we dry the precipitate under reduced pressure and mild conditions. In the event that zinc hyaluronate solution without ions is required, it is better to dissolve zinc hyaluronate, which does not contain solvents. With either of the two variants of the procedure, we get a hard substance without ions or a dissolved product with purity, which in the given example depends on the quality of the original zinc hyaluronate.
Rezultati kliničko-farmakoloških pretraga na sastojke (primjer 13), koji sadrže kao aktivni sastojak cinkov hijaluronat, prikazani su za liječenje ulkusa na bedru za ubrzavanje epitelizacije površina, gdje nedostaje epitel. Spoj, koji je sadržavao natrijev hijaluronat smo upotrijebili kao kontrolu. The results of clinical-pharmacological tests on the ingredients (example 13), which contain zinc hyaluronate as an active ingredient, are shown for the treatment of ulcers on the thigh to accelerate the epithelialization of surfaces where the epithelium is missing. We used the compound containing sodium hyaluronate as a control.
Tu pretragu smo izveli na 12 ili 14 ulkusa, odnosno na 8 ili 12 pacijenata, koji su bolovali od ulkusa na bedru. Podjela bolesnika obih skupina sa obzirom na spol i starost, kao i na prirodu oboljenja, bila je kako slijedi. We performed that search on 12 or 14 ulcers, that is, on 8 or 12 patients, who suffered from thigh ulcers. The distribution of patients in both groups with regard to gender and age, as well as the nature of the disease, was as follows.
[image] [image]
x A = arterijske; V = venske; M = miješane x A = arterial; V = venous; M = mixed
Liječenje smo izveli na takav način da smo prije početka liječenja izveli terapiju čišćenja sa obzirom na stvarno kliničko stanje ulkusa. Liječenje sa cinkovim ili natrijevim hijaluronatom smo počeli na netom očišćenim ulkusima ili u slučaju kada smo opazili značajno smanjenje sorpcije. Liječenje smo izvodili jedanput dnevno na takav način da smo pripravak nakapali na očišćenu površinu ulkusa u količini koja je namočila površinu rane u obliku tankog sloja. We carried out the treatment in such a way that before the start of the treatment, we carried out the cleaning therapy with regard to the actual clinical condition of the ulcer. We started treatment with zinc or sodium hyaluronate on freshly cleaned ulcers or in cases where we noticed a significant decrease in sorption. We performed the treatment once a day in such a way that we dripped the preparation onto the cleaned surface of the ulcer in an amount that soaked the surface of the wound in the form of a thin layer.
Pripravak smo upotrebljavali 4 tjedna. U početku liječenja i zatim jedanput tjedno ispunjavali smo formular i dokumentirali stanje ulkusa pacijenata sa fotografijama. Uzorak sekreta je bio uzet za bakteriološku pretragu. We used the preparation for 4 weeks. At the beginning of the treatment and then once a week, we filled out a form and documented the condition of the patients' ulcers with photographs. A secretion sample was taken for bacteriological examination.
Karakteristike kao i također jakost epitelijskih lezija su označeni sa slijedećim simbolima i vrijednostima. The characteristics as well as the strength of the epithelial lesions are marked with the following symbols and values.
Karakteristike Jakost Characteristics Strength
Površina (a) Area (a)
0 0 0 0
Ispod 10 cm2 1 Under 10 cm2 1
Između 10 cm2 i 25 cm2 2 Between 10 cm2 and 25 cm2 2
Iznad 25 cm2 3 Above 25 cm2 3
Infekcije (b) Infections (b)
Klinički čista 0 Clinically pure 0
Prekriveno u 50% 1 Covered in 50% 1
Prekriveno u 100% 2 Covered in 100% 2
Nekroza (c) Necrosis (c)
(samo u slučaju arterijskih ulkusa) (only in case of arterial ulcers)
negativno 0 negative 0
Ispod 10% 1 Below 10% 1
Između 10% i 15% 2 Between 10% and 15% 2
100% 3 100% 3
Bez nekroze 4 No necrosis 4
Ocjenjivanje gradeing
Ocijenili smo vrijednosti odvojenih karakteristika i izračunali vrijednost opće jakosti korištenjem slijedeće formule: We evaluated the values of the separate characteristics and calculated the value of the general strength using the following formula:
s = [image] s = [image]
Rezultati kliničko farmakoloških pretraga prikazani su na slici 2. Rezultati liječenja sa cinkovim hijaluronatom su prikazani na krivulji, označeni sa križićima, dok su rezultati liječenja sa natrijevim hijaluronatom prikazani na krivulji označeni sa kvadratićima, kao funkcija broja tjedana, koje je uključivalo liječenje. Vrijednosti su nacrtane na ordinati i predstavljaju opći indeks jakosti izračunat sa upotrebom gornje formule. The results of clinical pharmacological tests are shown in Figure 2. The results of treatment with zinc hyaluronate are shown on the curve, marked with crosses, while the results of treatment with sodium hyaluronate are shown on the curve marked with squares, as a function of the number of weeks, which included treatment. The values are plotted on the ordinate and represent the general strength index calculated using the above formula.
Za pravilniju usporedbu cinkovog hijaluronata sa natrijevim hijaluronatom, upotrijebljenim kao kontrola, pravilna je relativna vrijednost, koja se odnosi na ishodišne vrijednosti kao 100%, prikazana na slici 3. For a more correct comparison of zinc hyaluronate with sodium hyaluronate, used as a control, the correct relative value, which refers to the initial values as 100%, is shown in Figure 3.
Promjenu u relativno pravilnim vrijednostima statistički smo ocijenili kao funkciju broja (1-4) tjedana. Pri liječenju sa cinkovim i natrijevim hijaluronatom je broj ulkusa pao ispod relativne vrijednosti 90%, 80%, 70% i 60% nakon jednog, dva, tri i četiri tjedna pretraga. Rezultati su prikazani u tablici 1. We statistically evaluated the change in relatively regular values as a function of the number of (1-4) weeks. During treatment with zinc and sodium hyaluronate, the number of ulcers fell below the relative value of 90%, 80%, 70% and 60% after one, two, three and four weeks of examination. The results are presented in Table 1.
Tablica 1 Table 1
[image] [image]
Iz tablice 1 možemo zaključiti, da je liječenje sa cinkovim hijaluronatom u svakom tjednu bolje u odnosu na rezultate dobivene sa natrijevim hijaluronatom, koji je bio upotrebljen kao kontrola. From table 1, we can conclude that treatment with zinc hyaluronate in each week is better compared to the results obtained with sodium hyaluronate, which was used as a control.
Statistička analiza odgovora, dobivenog sa razmatranom hipotezom, je pokazala da je prednost spoja cinkovog hijaluronata vrlo signifikantna (p≥99%) u usporedbi sa natrijevim hijaluronatom. Statistical analysis of the response obtained with the considered hypothesis showed that the advantage of the zinc hyaluronate compound is very significant (p≥99%) compared to sodium hyaluronate.
Daljnja statistička obrada, detaljnije razdiobe i relativnih vrijednosti je bila istražena kao funkcija vremena liječenja. Dobiveni rezultati su prikazani u tablici 2. Further statistical processing, more detailed distributions and relative values were investigated as a function of treatment time. The obtained results are shown in table 2.
Tablica 2 Table 2
Broj i ocjena ulkusa Number and grade of ulcers
[image] [image]
Podaci iz tablice 2 slično podupiru prednost cinkovog hijaluronata. Detaljnija statistička pretraga pokazuje značajno smanjenje ovisno o vremenu liječenja. The data in Table 2 similarly support the benefit of zinc hyaluronate. A more detailed statistical analysis shows a significant decrease depending on the treatment time.
Ako sažmemo: ocjenjivanje kliničko farmakoloških pretraga je dokazalo visoku učinkovitost cinkovog hijaluronata čak pri manjem broju ulkusa; tu prednost još posebno podupire početno vrijeme liječenja. Izum je pravilnije objašnjen sa slijedećim primjerima, koji ga u nijednom pogledu ne ograničuju. If we summarize: the evaluation of clinical pharmacological tests has proven the high efficiency of zinc hyaluronate even with a smaller number of ulcers; this advantage is especially supported by the initial treatment time. The invention is more properly explained with the following examples, which do not limit it in any respect.
Vrijednost proteina hijaluronata (HA) je bila određena uporabom metode O. H. Lowry (J. Biol. Chem. 193 (1951)); viskoznost hijaluronata smo mjerili sa Ostwaldovim viskozimetrom u fiziološkoj otopini soli pri 25°C. Vrijednost za intrinzičku viskoznost ekstrapoliranu na koncentraciju "0" je dana ispod t.j. [η]c→0% 25 0 c. Sadržaj HA smo odredili uporabom The value of hyaluronate protein (HA) was determined using the method of O. H. Lowry (J. Biol. Chem. 193 (1951)); the viscosity of hyaluronate was measured with an Ostwald viscometer in physiological salt solution at 25°C. The value for intrinsic viscosity extrapolated to "0" concentration is given below i.e. [η]c→0% 25 0 c. We determined the content of HA by using it
Bitterijeve metode (Anal. Biochem 4, 330 (1962)). Bitteri's methods (Anal. Biochem 4, 330 (1962)).
Primjer 1 Example 1
Priprema otopine cinkovog hijaluronata Preparation of zinc hyaluronate solution
40,18 mg natrijevog hijaluronata smo rastopili u 20,0 ml dva puta destilirane vode. Tako je bila ishodišna koncentracija hijaluronske kiseline 2,009 mg/ml, ekvivalentna koncentraciji otopine 4.241x10-3 mola/l (Na+ ili dimera hijaluronske kiseline). U postupku mjerenja smo dodali otopinu cinkovog klorida koncentracije 0.05154 mol/l u reakcijsku smjesu pomoću mikrobirete. Otopinu smo najprije dodavali u malim dijelovima (0,05 ml) i zatim u većim dijelovima (od 0,1 do 0,2 ml). Promjenu potencijala otopine smo mjerili uporabom osjetljivog potenciometra sa digitalnim prikazom i natrijeve ion-selektivne elektrode i elektrode srebro/srebreni klorid. Titraciju smo nastavili dok se mjereni potencijal nije više mijenjao pri dodavanju dodatnih dijelova titrirne otopine. (Mjerni sustav smo kalibrirali pri uvjetima sličnim praktičnim mjerenjima). We dissolved 40.18 mg of sodium hyaluronate in 20.0 ml of twice-distilled water. Thus, the initial concentration of hyaluronic acid was 2.009 mg/ml, equivalent to the concentration of the solution 4.241x10-3 mol/l (Na+ or hyaluronic acid dimer). In the measurement process, we added a solution of zinc chloride with a concentration of 0.05154 mol/l to the reaction mixture using a microburette. The solution was first added in small portions (0.05 ml) and then in larger portions (from 0.1 to 0.2 ml). We measured the change in potential of the solution using a sensitive potentiometer with a digital display and a sodium ion-selective electrode and a silver/silver chloride electrode. We continued the titration until the measured potential did not change any more when adding additional parts of the titration solution. (We calibrated the measuring system under conditions similar to practical measurements).
Selektivnost natrij ion-selektivne elektrode smo opazili također u prisutnosti Zn2+ iona, da bi kontrolirali, da je promjena potencijala u praktičnim mjerenjima uzrokovana oslobađanjem Na+ iona a ne Zn2+ iona, koje smo uveli u otopinu. 2.00x10-3 M otopinu natrijevog klorida smo titrirali uporabom otopine cinkovog klorida pri uvjetima, koji su bili slični gornjim uvjetima. Pri rastu koncentracije Zn2+ od 0 do 4x10-3 mola/I opazili smo porast potencijala za oko 2 mV, dok su praktična mjerenja pokazala promjene oko 20 mV pri istim uvjetima. Takvo ocjenjivanje nije bilo ometano. We observed the selectivity of the sodium ion-selective electrode also in the presence of Zn2+ ions, in order to control that the potential change in practical measurements was caused by the release of Na+ ions and not Zn2+ ions, which we introduced into the solution. A 2.00x10-3 M sodium chloride solution was titrated using a zinc chloride solution under conditions similar to the above conditions. When the concentration of Zn2+ increased from 0 to 4x10-3 mol/I, we observed an increase in potential by about 2 mV, while practical measurements showed changes of about 20 mV under the same conditions. Such evaluation was not hindered.
U postupku mjerenja je porast aktivnosti natrijevog iona izračunat iz podataka mjerenja dokazao kavantitativni nastanak cinkovog kompleksa. During the measurement process, the increase in sodium ion activity calculated from the measurement data proved the quantitative formation of the zinc complex.
Priprema otopine cinkovog klorida Preparation of zinc chloride solution
Kako otopinu koja sadrži cinkov klorid nije bilo moguće pripraviti u točnoj koncentraciji direktnim vaganjem, najprije smo pripravili otopinu sa približno željenom koncentracijom. Pri pripremanju te otopine nismo smjeli uporabiti kiselinu, jer bi se moglo dogoditi da se izvagani cinkov klorid ne bi potpuno otopio. Po sedimentaciji netopljivog ostatka (oko 30 minuta) bila je volumetrijska posuda napunjena do oznake a otopina filtrirana kroz papir za filtriranje. Since it was not possible to prepare a solution containing zinc chloride in the exact concentration by direct weighing, we first prepared a solution with approximately the desired concentration. When preparing this solution, we should not have used acid, because it could happen that the weighed zinc chloride would not dissolve completely. After sedimentation of the insoluble residue (about 30 minutes), the volumetric container was filled to the mark and the solution was filtered through filter paper.
Točnost koncentracije fiitrata odredili smo sa kompleksometričnom titracijom sa uporabom pufera 10 i eriokrom crnog T kao indikatora. Otopinu cinkovog klorida sa točnom koncentracijom 0,100 mola/I pripravili smo sa točnim razrjeđivanjem te otopine. We determined the accuracy of the nitrate concentration with complexometric titration using buffer 10 and eriochrome black T as an indicator. A solution of zinc chloride with an exact concentration of 0.100 mol/I was prepared with the exact dilution of this solution.
Karakteristike natrijevog hijaluronata upotrijebljenog za pripravljanje te otopine su kako slijedi: The characteristics of sodium hyaluronate used for the preparation of this solution are as follows:
Molska masa : 1850000 daltona Molar mass: 1850000 daltons
Sadržj proteina : 0,07 % Protein content: 0.07%
UV apsorpcija A1%257 : 0,133 UV absorption A1%257 : 0.133
A1%280 : 0,075 A1%280 : 0.075
Viskoznost [η]c→0% 250c : 13,7 dl/g Viscosity [η]c→0% 250c: 13.7 dl/g
Sadržaj HAx: 98,12 % HAx content: 98.12%
xHA = hijaluronska kiselina (kako je to skraćeno na tom mjestu) xHA = Hyaluronic Acid (as abbreviated here)
Primjer 2 Example 2
Priprema otopine za dermatološku i kozmetičku upotrebu Preparation of solutions for dermatological and cosmetic use
12,5 ml otopine cinkovog klorida sa konc. 0,100 mol/I pripravili smo sa dejoniziranom vodom i dodali u 0,50 g natrijevog hijaluronata izvaganog u 100 ml volumetriskoj posudi. (Možemo uportrijebiti također drugu koncentraciju cinkovog klorida ali količina cinkovog klorida mora biti jednaka). Natrijev hijaluronat smo pustili bubriti (za 12 sati) i otopinu dopunili do oznake za dejoniziranu vodu, da bi dobili otopinu cinkovog hijaluronata 0,5 mas. %./vol. Karakteristike upotrebljenog natrijevog hijaluronata za pripravljanje gornje otopine su kako slijedi: 12.5 ml of zinc chloride solution with conc. 0.100 mol/I was prepared with deionized water and added to 0.50 g of sodium hyaluronate weighed in a 100 ml volumetric container. (We can also use another concentration of zinc chloride, but the amount of zinc chloride must be the same). We let the sodium hyaluronate swell (for 12 hours) and topped up the solution to the mark for deionized water, in order to obtain a solution of zinc hyaluronate 0.5 wt. %./vol. The characteristics of the sodium hyaluronate used for the preparation of the above solution are as follows:
Viskoznost [η]c→0% 250c : 16,5 dl/g Viscosity [η]c→0% 250c: 16.5 dl/g
Sadržaj proteina : 0,8 mas.% Protein content: 0.8 wt.%
Primjer 3 Example 3
Priprema otopine cinkovog hijaluronata za uporabu u injekcijskim otopinama. Preparation of zinc hyaluronate solution for use in injection solutions.
Operacije opisane u ovom primjeru su bile izvedene u sterilnim uvjetima. The operations described in this example were performed under sterile conditions.
Otopinu 5,0 ml cinkovog klorida koncentracije 0,100 mola/I pripravljene sa dva puta destiliranom vodom (voda za uporabu u injekcijama, bez pirogena, sterilna) smo dodali u 0,20 g natrijevog hijaluronata (čistoće kvalitete čistoće praška) izvaganog u 100 ml volumetrijskoj posudi, zatim smo napunili do volumena 50 ml sa dva puta destiliranom vodom. A solution of 5.0 ml of zinc chloride with a concentration of 0.100 mol/L prepared with twice-distilled water (water for use in injections, without pyrogens, sterile) was added to 0.20 g of sodium hyaluronate (purity of powder quality) weighed in a 100 ml volumetric container, then we filled it up to a volume of 50 ml with twice-distilled water.
Natrijev hijaluronat smo ostavili bubriti preko noći, nakon čega smo ga otopili uz trešnju i otopinu dopunili do oznake sa dva puta destiliranom vodom. Dobivenu otopinu smo filtrirali kroz filtarsku membranu (0,45 μ veličine pora) da bi dobili otopinu cinkovog hijaluronata 0,2 mas. %/vol. Karakteristike upotrebljenog natrijevog hijaluronata za pripravu gornjih otopina su kako slijedi: We left the sodium hyaluronate to swell overnight, after which we dissolved it with the cherry and topped up the solution to the mark with twice-distilled water. The resulting solution was filtered through a filter membrane (0.45 μ pore size) to obtain a solution of zinc hyaluronate 0.2 wt. %/vol. The characteristics of the sodium hyaluronate used for the preparation of the above solutions are as follows:
Kvaliteta: čist, bez pirogena, sterilan prašak Quality: pure, pyrogen-free, sterile powder
Molska masa: 1850000 Molar mass: 1850000
Sadržaj proteina: 0,07 mas.% Protein content: 0.07 wt.%
UV apsorpcija A1%257 0,133 UV absorption A1%257 0.133
A1%280 0,075 A1%280 0.075
Sadržaj HA: 98,12 mas.% HA content: 98.12 wt.%
Viskoznost [η]c→0% 250c 13,7 dl/g Viscosity [η]c→0% 250c 13.7 dl/g
Primjer 4 Example 4
Priprema otopine cinkovog hijalorunata bez iona Preparation of zinc hyaluronate solution without ions
600 ml etanola analitičke čistoće dodamo među miješanjem u 200 ml 0,50 mas. %/vol. otopine cinkovog hijaluronata dobivene prema primjeru 2, istaložen cinkov hijaluronat filtriramo na staklenom filtru, isperemo svaki put po dva puta sa 50 ml etanola, jednake kvalitete i zatim sušimo pod smanjenim tlakom. Tako smo dobili 0,88 g cinkovog hijaluronata, kojeg smo za pripravu uporabili 0,50 mas. %/vol. otopine cinkovog hijaluronata na način opisan u primjeru 2. Dobivena otopina cinkovog hijaluronata nije sadržavala ništa natrijevog klorida, koji se pojavi pri reakciji između natrijevog hijaluronata i cinkovog klorida; tako je praktično bez iona. Add 600 ml of ethanol of analytical purity to 200 ml of 0.50 wt. %/vol. zinc hyaluronate solution obtained according to example 2, the precipitated zinc hyaluronate is filtered on a glass filter, washed each time twice with 50 ml of ethanol, of the same quality, and then dried under reduced pressure. Thus, we obtained 0.88 g of zinc hyaluronate, of which we used 0.50 wt. %/vol. zinc hyaluronate solution in the manner described in example 2. The resulting zinc hyaluronate solution did not contain any sodium chloride, which appears during the reaction between sodium hyaluronate and zinc chloride; it is so practically ion-free.
Primjer 5 Example 5
Priprema cinkovog hijaluronata ili njegove otopine bez iona za terapijsku uporabu. Preparation of zinc hyaluronate or its ion-free solution for therapeutic use.
Operacije, opisane u ovom primjeru izveli smo u sterilnim uvjetima. The operations described in this example were performed under sterile conditions.
1500 ml etanola (najčišće kvalitete) smo u dijelovima dodavali otopini 500 ml cinkovog hijaluronata, pripravljene prema primjeru 3 uz miješanje. Po dodavanju smo sistem miješali daljnjih 30 minuta, istaloženi cinkov hijaluronat filtrirali na staklenom filtru, isprali tri puta, svaki put sa 100 ml etanola (najčišće kvalitete) i sušili pod smanjenim tlakom u blagim i sterilnim uvjetima. 1500 ml of ethanol (of the purest quality) was added in parts to a solution of 500 ml of zinc hyaluronate, prepared according to example 3, while stirring. After addition, the system was stirred for another 30 minutes, the precipitated zinc hyaluronate was filtered on a glass filter, washed three times, each time with 100 ml of ethanol (the purest quality) and dried under reduced pressure in mild and sterile conditions.
Primjer 6 Example 6
Priprema cinkovog hijaluronata bez iona. Preparation of zinc hyaluronate without ions.
200 ml 10 mas.% otopine hiamina HyamineR 1622 (puriss) (benzildimetil/-2-/2-p-(1,1,3,3-tetrametil butil) fenoksi/ etoksi/etil/amonijev klorid) dodajemo uz miješanje otopini koja sadrži 1 g natrijevog hijaluronata u 400 ml dva puta destilirane vode. Talog, t.j. nastali kompleks hijalurinske kiseline sa kvatemarnim amonijakom odvojimo i ponovno centrifugiramo. Isprani talog otopimo u paru otapala, koji se sastoji iz 400 ml 2 mas.%/vol. cinkovog klorida i vodene otopine (pH 5,0 do 5,4) i 400 ml n-butanola. Kada se obe faze odvoje, vodeni sloj, koji sadrži otopljen cinkov hijaluronat filtriramo kroz membranski filtar (0,45 μ veličine pora), zatim istaložimo cinkov hijaluronat sa dodatkom tri volumena etanola, filtriramo na staklenom filteru, isperemo sa etanolom i sušimo u dušikovoj atmosferi pod blagim uvjetima, da dobijemo 0,82 g cinkovog hijaluronata. 200 ml of 10 wt.% hyamine solution HyamineR 1622 (puriss) (benzyldimethyl/-2-/2-p-(1,1,3,3-tetramethyl butyl)phenoxy/ethoxy/ethyl/ammonium chloride) is added with stirring to the solution that contains 1 g of sodium hyaluronate in 400 ml of twice distilled water. Sediment, i.e. the resulting complex of hyaluronic acid with quaternary ammonia is separated and centrifuged again. Dissolve the washed precipitate in solvent vapor, which consists of 400 ml of 2 wt.%/vol. zinc chloride and aqueous solution (pH 5.0 to 5.4) and 400 ml of n-butanol. When both phases are separated, the aqueous layer, which contains dissolved zinc hyaluronate, is filtered through a membrane filter (0.45 μ pore size), then the zinc hyaluronate is precipitated with the addition of three volumes of ethanol, filtered on a glass filter, washed with ethanol and dried in a nitrogen atmosphere under mild conditions, to obtain 0.82 g of zinc hyaluronate.
Ako je potrebno iz dobivenog cinkovog hijaluronata, pripravljamo otopinu 0,50 mas.%/vol, koju zatim dalje čistimo kako je opisano u primjeru 4. Karakteristike upotrebljenog natrijevog hijaluronata kao ishodične tvari su kako slijedi: If necessary, we prepare a 0.50 wt.%/vol solution from the obtained zinc hyaluronate, which is then further purified as described in example 4. The characteristics of the sodium hyaluronate used as a starting material are as follows:
Viskoznost [η]c→0% 250c : 16,5 dl/g Viscosity [η]c→0% 250c: 16.5 dl/g
Sadržaj proteina : 0,18 mas.%/vol. Protein content: 0.18 wt.%/vol.
Cinkov hijaluronat možemo pripraviti kako je gore opisano također iz kompleksa koji nastaje iz drugih kvaternarnih amonijevih soli. Za tu svrhu upotrebive kvaternarne soli su npr.: Zinc hyaluronate can be prepared as described above also from the complex formed from other quaternary ammonium salts. Quaternary salts that can be used for this purpose are, for example:
a) karbotetradeciloksimetil-trimetilamonijev klorid a) carbotetradecyloxymethyl-trimethylammonium chloride
(vidi Mađarski patentni spis broj 188,537), (see Hungarian patent file number 188,537),
b) heksadecilpiridinijev klorid, b) hexadecylpyridinium chloride,
c) cetilpiridinijev klorid, c) cetylpyridinium chloride,
d) trimetilamonijev klorid i slično. d) trimethylammonium chloride and the like.
Primjer 7 Example 7
Priprema kobaltnog hijaluronata Preparation of cobalt hyaluronate
Slijedimo postupak opisan u primjeru 6, samo što kompleks hijaluronske kiseline kvaternarnog amonijaka otopimo u paru otapala, kojega čine 2 mas.%/vol. vodena otopina kobaltovog (II) klorida.6H2O i n-butanol. We follow the procedure described in example 6, except that we dissolve the quaternary ammonia hyaluronic acid complex in solvent vapor, which is 2 wt.%/vol. aqueous solution of cobalt (II) chloride.6H2O and n-butanol.
Primjer 8 Example 8
Priprema vodene otopine, koja sadrži 0,50 mas.%/vol. cinkovog hijaluronata, koju napravimo izotoničnom sa cinkovim kloridom. Preparation of an aqueous solution, which contains 0.50 wt.%/vol. of zinc hyaluronate, which we make isotonic with zinc chloride.
Oko 50 ml otopine cinkovog klorida koncentracije 0,110 mola/I dodamo u 0,50 g natrijevog hijaluronata u 100 ml volumetrijsku posudu i zatim ostavimo bubriti preko noći. Nakon toga otopimo natrijev hijaluronat sa trešnjom i posudu napunimo do oznake sa otopinom cinkovog klorida koncentracije 0,110 mola/I. Add about 50 ml of zinc chloride solution with a concentration of 0.110 mol/I to 0.50 g of sodium hyaluronate in a 100 ml volumetric container and then leave it to swell overnight. After that, dissolve the sodium hyaluronate with the cherry and fill the container up to the mark with a zinc chloride solution with a concentration of 0.110 mol/I.
Dobiven osmotski tlak otopine 0,1491 mola/I je izražen kao ekvivalent koncentracije natrijevog klorida pH vrijednosti 5,0. Ako je potrebno poravnamo pH vrijednost od 5,5 do 5,6 sa dodatkom 2,00 ml otopine natrijevog acetata koncentracije 0,150 mola/I. Nakon poravnanja pH vrijednosti osmotski je tlak otopine 0,1489 izražen kao ekvivalent koncentracije natrijevog klorida. The resulting osmotic pressure of the solution of 0.1491 mol/I is expressed as the sodium chloride concentration equivalent of a pH value of 5.0. If necessary, adjust the pH value from 5.5 to 5.6 with the addition of 2.00 ml of sodium acetate solution with a concentration of 0.150 mol/I. After adjusting the pH value, the osmotic pressure of the solution is 0.1489, expressed as the equivalent concentration of sodium chloride.
Otopinu cinkovog hijaluronata pripremamo iz posebno čistog natrijevog hijaluronata opisanog u primjeru 3 sa dva puta destiliranom vodom u aseptičkim uvjetima, zatim otopinu filtriramo kroz membranski filtar (0,65 μ veličine pora). Otopinu možemo upotrijebiti također u injekcijskim pripravcima. The zinc hyaluronate solution is prepared from the especially pure sodium hyaluronate described in example 3 with twice-distilled water under aseptic conditions, then the solution is filtered through a membrane filter (0.65 μ pore size). The solution can also be used in injectable preparations.
Primjer 9 Example 9
Priprema vodene otopine, koja sadrži 0,2 mas.%/vol. cinkovog hijaluronata pripravljena izotonično sa cinkovim kloridom. Preparation of an aqueous solution, which contains 0.2 wt.%/vol. of zinc hyaluronate prepared isotonically with zinc chloride.
Za konačni volumen 100 ml smo izvagali 0,20 g natrijevog hijaluronata i otopili u otopini cinkovog klorida koncentracije 0,120 mola/I. For the final volume of 100 ml, we weighed 0.20 g of sodium hyaluronate and dissolved it in a zinc chloride solution with a concentration of 0.120 mol/L.
Otapljanje i pripravu otopine cinkovog klorida konc. točno 0,120 mola/I smo izveli prema primjeru 1 (sa time da smo promijenili količinu cinkovog klorida). Dissolving and preparing a solution of zinc chloride conc. we derived exactly 0.120 mol/I according to example 1 (with the fact that we changed the amount of zinc chloride).
Osmotski tlak otopine je 0,154 mola/I izražen kao ekvivalent koncentracije natrijevog klorida; pH pokazuje vrijednost od 5,3 do 5,4. The osmotic pressure of the solution is 0.154 mol/I expressed as the equivalent concentration of sodium chloride; The pH shows a value of 5.3 to 5.4.
Sadržaj HA : 1,96 mg/ml HA content: 1.96 mg/ml
Primjer 10 Example 10
Priprema vodene otopine, koja sadrži 0,2% mas.%/vol. cinkovog hijaluronata pripravljene izotonično sa glukozom. Preparation of an aqueous solution, which contains 0.2% wt.%/vol. zinc hyaluronate prepared isotonically with glucose.
Otopina ovog primjera sadrži natrijev hijaluronat i izračunatu ekvivalentnu količinu cinkovog klorida. The solution of this example contains sodium hyaluronate and a calculated equivalent amount of zinc chloride.
5,0 ml otopine cinkovog klorida koncentracije 0,100 mola/I dodamo 0,20 g izvaganog natrijevog hijaluronata u 100 ml volumetrijsku posudu, zatim volumen dopunimo do 50 ml sa dejoniziranom vodom. Nakon što smo ostavili bubriti preko noći natrijev hijaluronat otopimo stresanjem i dodamo 27,0 ml otopine glukoze koncentracije 1,00 mola/I i zatim dopunimo posudu do oznake sa dejoniziranom vodom. Add 0.20 g of weighed sodium hyaluronate to 5.0 ml of zinc chloride solution with a concentration of 0.100 mol/L in a 100 ml volumetric container, then make the volume up to 50 ml with deionized water. After we have left it to swell overnight, dissolve the sodium hyaluronate by shaking and add 27.0 ml of glucose solution with a concentration of 1.00 mol/I and then fill the container up to the mark with deionized water.
Osmotski tlak otopine je 0,151 mola/I, izražen kao ekvivalent koncentracije natrijevog klorida; pH pokazuje vrijednost od 5,6 do 5,7; cjelokupna koncentracija cinka je 5 x 10-3 mola/I. The osmotic pressure of the solution is 0.151 mol/I, expressed as the equivalent concentration of sodium chloride; pH shows a value of 5.6 to 5.7; the overall concentration of zinc is 5 x 10-3 mol/I.
Primjer 11 Example 11
Priprema vodene otopine, koja sadrži 0,5 mas.%/vol. cinkovog hijaluronata pripravljene izotonično sa sorbitolom. Preparation of an aqueous solution, which contains 0.5 wt.%/vol. zinc hyaluronate prepared isotonically with sorbitol.
Dolje opisanu otopinu cinkovog hijaluronata smo pripravili u aseptičnim uvjetima iz natrijevog hijaluronata posebno visoke čistoće, opisanog u primjeru 3 i sa destiliranom vodom. Otopina sadrži cinkov klorid u ekvivalentnoj količini, izračunatoj za natrijev hijaluronat. Slijedili smo postupak, opisan u primjeru 5, ali smo umjesto otopine glukoze dodali 23,50 ml otopine sorbitola, koncentracije 1,00 mola/I (182,19 g D-sorbitola u 1 litru), otopini cinkovog hijaluronata. The zinc hyaluronate solution described below was prepared under aseptic conditions from sodium hyaluronate of particularly high purity, described in example 3, and with distilled water. The solution contains zinc chloride in an equivalent amount calculated for sodium hyaluronate. We followed the procedure described in example 5, but instead of the glucose solution, we added 23.50 ml of sorbitol solution, concentration 1.00 mol/L (182.19 g of D-sorbitol in 1 liter), to the zinc hyaluronate solution.
Tako pripravljenu otopinu smo filtrirali kroz membranski filtar (0,45 μ veličine pora). Tu otopinu možemo upotrijebiti u bilo koju svrhu, koja uključuje injekcijske pripravke. The solution thus prepared was filtered through a membrane filter (0.45 μ pore size). We can use this solution for any purpose, which includes injectable preparations.
Viskoznost : 15,9 dl/g Viscosity: 15.9 dl/g
Sadržaj proteina : 0,015 mg/ml Protein content: 0.015 mg/ml
Čistoća otopinex : A1%660 = 0,015 Solution purityx : A1%660 = 0.015
xNa osnovi apsorpcije mjerene pri 660 nm u 1 cm kiveti. xBased on absorbance measured at 660 nm in a 1 cm cuvette.
Otopinu smo pripravili uporabom natrijevog hijaluronata kvalitete označene u primjeru 2 i zatim ju uporabili za pripravu dermatoloških i kozmetičkih pripravaka. We prepared the solution using sodium hyaluronate of the quality indicated in example 2 and then used it for the preparation of dermatological and cosmetic preparations.
Primjer 12 Example 12
Priprema vodene otopine, koja sadrži 0,50 mas.%/vol. cinkovog hijaluronata, pripravljene izotonično sa glukozom. Preparation of an aqueous solution, which contains 0.50 wt.%/vol. zinc hyaluronate, prepared isotonically with glucose.
Otopina ovog primjera sadrži natrijev hijaluronat i izračunatu ekvivalentnu količinu cinkovog klorida. The solution of this example contains sodium hyaluronate and a calculated equivalent amount of zinc chloride.
12,50 ml cinkovog klorida konc. 0,100 mola/l dodamo 0,50 g izvaganog natrijevog hijaluronata u 100 ml volumetrijsku posudu. 12.50 ml zinc chloride conc. 0.100 mol/l, add 0.50 g of weighed sodium hyaluronate to a 100 ml volumetric container.
(Možemo upotrijebiti također drugačiju koncentraciju cinkovog klorida ali mora biti količina cinkovog klorida jednaka). Natrijev hijaluronat ostavimo bubriti 12 sati i otopinu cinkovog klorida dopunimo do 50 ml sa dejoniziranom vodom, i zatim otopimo stresanjem. Nakon toga dodamo 24,50 ml otopine glukoze koncentracije 1,00 mola/I i dopunimo do oznake sa dejoniziranom vodom. (We can also use a different concentration of zinc chloride, but the amount of zinc chloride must be the same). Leave the sodium hyaluronate to swell for 12 hours and top up the zinc chloride solution to 50 ml with deionized water, and then dissolve by shaking. After that, add 24.50 ml of glucose solution with a concentration of 1.00 mol/L and top up to the mark with deionized water.
Osmotski tlak otopine je 0,1495 mola/I izražen kao ekvivalent koncentracije natrijevog klorida; pH pokazuje vrijednost 5,4. Cjelokupna koncentracija cinka je 1,25x10-2 mola/l. The osmotic pressure of the solution is 0.1495 mol/I expressed as the equivalent concentration of sodium chloride; The pH shows a value of 5.4. The total concentration of zinc is 1.25x10-2 mol/l.
Otopinu pripravljamo sa pripravom natrijevog hijaluronata kvalitete označene u primjeru 2 i koristimo ju najprije za pripravu dermatoloških i kozmetičkih pripravaka. We prepare the solution with sodium hyaluronate of the quality indicated in example 2 and use it first for the preparation of dermatological and cosmetic preparations.
Osmotski tlak otopine je 0,1520 mola/I izražen kao ekvivalent koncentracije natrijevog klorida; pH pokazuje vrijednost oko 5,5; cjelokupna koncentracija je 1,25x10-2 mola/I. The osmotic pressure of the solution is 0.1520 mol/I expressed as the equivalent concentration of sodium chloride; pH shows a value of around 5.5; the overall concentration is 1.25x10-2 mol/I.
Primjer 13 Example 13
Priprema vodene otopine, koja sadrži 0,2 mas.%/vol. cinkovog hijaluronata pripravljene izotonično sa sorbitolom Preparation of an aqueous solution, which contains 0.2 wt.%/vol. zinc hyaluronate prepared isotonically with sorbitol
Otopina, opisana u ovom primjeru, sadrži cinkov klorid u ekvivalentnoj količini, izračunatoj za natrijev hijaluronat. The solution described in this example contains zinc chloride in an equivalent amount calculated for sodium hyaluronate.
Niže opisanu otopinu cinkovog hijaluronata smo pripremili u aseptičkim uvjetima iz natrijevog hijaluronata posebno visoke čistoće, opisanog u primjeru 3 sa dva puta destiliranom vodom. Slijedili smo postupak primjera 12, samo što smo otopili 0,2 g natrijevog hijaluronata, 5 ml otopine cinkovog klorida, koncentracije 0,100 mola/I i na to dodali 26,50 ml otopine sorbitola u konc. 1 mol/I i konačno otopinu dopunili do 100 ml. Tako pripravljenu otopinu smo filtrirali kroz membranski filtar (0,45 μ veličine pora). Tu otopinu možemo upotrijebiti u bilo koje svrhe, koje uključuju injekcijske pripravke. The zinc hyaluronate solution described below was prepared under aseptic conditions from sodium hyaluronate of particularly high purity, described in example 3, with twice-distilled water. We followed the procedure of example 12, except that we dissolved 0.2 g of sodium hyaluronate, 5 ml of zinc chloride solution, concentration 0.100 mol/I and added 26.50 ml of sorbitol solution in conc. 1 mol/I and finally the solution was made up to 100 ml. The solution thus prepared was filtered through a membrane filter (0.45 μ pore size). We can use this solution for any purpose, which includes injectable preparations.
Osmotski tlak otopine je 0,1401 mola/I i izražen kao ekvivalent koncentracije natrijevog klorida; pH pokazuje vrijednost oko 5,6; cjelokupna koncentracija cinka je 5x10-3 mola/I. The osmotic pressure of the solution is 0.1401 mol/I and is expressed as the equivalent concentration of sodium chloride; pH shows a value of around 5.6; the overall concentration of zinc is 5x10-3 mol/I.
Sadržaj hijaluronata : 2,03 mg/ml Hyaluronate content: 2.03 mg/ml
Viskoznost : 16,1 dl/g Viscosity: 16.1 dl/g
Sadržaj proteina : 0,016 mg/ml Protein content: 0.016 mg/ml
Čistoća otopinexA1cm660 = 0,010 Solution purityxA1cm660 = 0.010
xNa osnovi absopcije, mjerene pri 660 nm u 1 cm kiveti. xBased on absorbance, measured at 660 nm in a 1 cm cuvette.
Primjeri 14 do 26 Examples 14 to 26
U slijedećim primjerima su dane komponente različitih pripravaka (farmaceutskih i kozmetičkih pripravaka) u svezi sa tipovima formulacija, koje smo izabrali. Priprave otopina cinkovog hijaluronata, pripravljenih izotonično, opisane su u prethodnim primjerima. Na ovom mjestu "destilirana voda za injekcijske namjene" znači dva puta destiliranu vodu pripravljenu u aseptičkim uvjetima. In the following examples, the components of various preparations (pharmaceutical and cosmetic preparations) are given in connection with the types of formulations that we have chosen. Preparations of zinc hyaluronate solutions, prepared isotonically, are described in the previous examples. In this context, "distilled water for injection" means doubly distilled water prepared under aseptic conditions.
I. Injekcijske otopine I. Injection solutions
Upotrijebili smo pripravke primjera 14 do 17 za intrakutano davanje, dok je onaj iz primjera 18 služio za intraokularno upotrebu. U tim primjerima smo upotrijebili aktivni sastojak kvalitete, opisane u primjeru 3. We used the preparations of examples 14 to 17 for intracutaneous administration, while the one from example 18 was for intraocular use. In these examples, we used the active ingredient of the quality described in example 3.
Primjer 14 Example 14
Cinkov hijaluronat kao aktivni sastojak 2,0 mg Zinc hyaluronate as active ingredient 2.0 mg
Sorbitol 48,3 mg Sorbitol 48.3 mg
Konačni volumen vodene otopine, pripravljene The final volume of the aqueous solution, prepared
sa destiliranom vodom za injekcijske namjene 1,0 ml with distilled water for injection purposes 1.0 ml
Primjer 15 Example 15
Cinkov hijaluronat kao aktivni sastojak 5,0 ml Zinc hyaluronate as active ingredient 5.0 ml
Sorbitol 42,8 mg Sorbitol 42.8 mg
Konačni volumen vodene otopine, pripravljene The final volume of the aqueous solution, prepared
sa destiliranom vodom za injekcijske namjene 1,0 ml with distilled water for injection purposes 1.0 ml
Primjer 16 Example 16
Cinkov hijaluronat kao aktivni sastojak 2,0 mg Zinc hyaluronate as active ingredient 2.0 mg
Propil p-hidroksibenzoat 0,05 mg Propyl p-hydroxybenzoate 0.05 mg
Metil p-hidroksibenzoat 0,5 mg Methyl p-hydroxybenzoate 0.5 mg
Glukoza 48,6 mg Glucose 48.6 mg
Konačni volumen vodene otopine, pripravljen The final volume of the aqueous solution, prepared
sa destiliranom vodom za injekcijske namjene 1,0 ml with distilled water for injection purposes 1.0 ml
Primjer 17 Example 17
Cinkov hijaluronat kao aktivni sastojak 5,0 mg Zinc hyaluronate as active ingredient 5.0 mg
Propil p-hidroksibenzoat 0,05 mg Propyl p-hydroxybenzoate 0.05 mg
Metil p-hidroksibenzoat 0,5 mg Methyl p-hydroxybenzoate 0.5 mg
Glukoza 44,1 mg Glucose 44.1 mg
Konačni volumen vodene otopine, pripravljen sa The final volume of the aqueous solution, prepared with
destiliranom vodom za injekcijske namjene 1,8 ml with distilled water for injection purposes 1.8 ml
Primjer 18 Example 18
Cinkov hijaluronat kao aktivni sastojak 10,0 mg Zinc hyaluronate as active ingredient 10.0 mg
Kalijev sorbat 1,0 mg Potassium sorbate 1.0 mg
Sorbitol 41,0 mg Sorbitol 41.0 mg
Konačni volumen vodene otopine, pripravljen sa The final volume of the aqueous solution, prepared with
destiliranom vodom za injekcijske namjene 1,0 ml with distilled water for injection purposes 1.0 ml
Pripravci opisani u primjeru 20 do 28 su uglavnom upotrebljeni u dermatološke i kozmetičke svrhe. Aktivni sastojci, koje smo upotrijebili u tim primjerima su kvalitete, opisane u primjeru 2. The preparations described in examples 20 to 28 are mainly used for dermatological and cosmetic purposes. The active ingredients used in these examples are of the quality described in example 2.
II Otopine za vanjsku primjenu II Solutions for external use
Primjer 19 Example 19
Cinkov hijaluronat kao aktivni sastojak 5,0 mg Zinc hyaluronate as active ingredient 5.0 mg
Kalijev sorbat 1,0 mg Potassium sorbate 1.0 mg
Natrijev acetat 24,6 mg Sodium acetate 24.6 mg
Konačni volumen vodene otopine, pripravljen The final volume of the aqueous solution, prepared
sa destiliranom vodom 1,0 ml with distilled water 1.0 ml
Primjer 20 Example 20
Cinkov hijaluronat kao aktivni sastojak 2,0 mg Zinc hyaluronate as active ingredient 2.0 mg
Kalijev sorbat 1,0 mg Potassium sorbate 1.0 mg
Sorbitol 48,3 mg Sorbitol 48.3 mg
Konačni volumen vodene otopine, pripravljen The final volume of the aqueous solution, prepared
sa destiliranom vodom 1,0 ml with distilled water 1.0 ml
III. Gelovi za lokalnu upotrebu III. Gels for local use
Primjer 21 Example 21
Cinkov hijaluronat kao aktivni sastojak 20,0 mg Zinc hyaluronate as active ingredient 20.0 mg
Polimerazit akrilne kiseline 200 mg Acrylic acid polymer 200 mg
Natrijev hidroksid 30%-ne koncentracije 50 mg Sodium hydroxide 30% concentration 50 mg
Kalijev sorbat 10 mg Potassium sorbate 10 mg
Destilirana voda do 10,0 mg Distilled water up to 10.0 mg
Primjer 22 Example 22
Cinkov hijaluronat kao aktivni sastojak 20,0 mg Zinc hyaluronate as active ingredient 20.0 mg
Polimerizat akrilne kiseline 50 mg Acrylic acid polymer 50 mg
Natrijev hidroksid 30%-ne koncentracije 40 mg Sodium hydroxide 30% concentration 40 mg
Propilen glikol 1500 mg Propylene glycol 1500 mg
Kalijev sorbat 10 mg Potassium sorbate 10 mg
Destilirana voda do 10,0 mg Distilled water up to 10.0 mg
IV. Kreme i masti za lokalnu upotrebu IV. Creams and ointments for local use
Primjer 23 Example 23
Cinkov hijaluronat kao aktivni sastojak 50 mg Zinc hyaluronate as an active ingredient 50 mg
Kalijev sorbat 10 mg Potassium sorbate 10 mg
Mekani, bijeli pčelinji vosak 125 mg Soft, white beeswax 125 mg
Sorbitan oleat 150 mg Sorbitan oleate 150 mg
Cetil stearil alkohol 840 mg Cetyl stearyl alcohol 840 mg
Gliceril monostearat 1100 mg Glyceryl monostearate 1100 mg
Propilen glikol 4750 mg Propylene glycol 4750 mg
Destilirana voda do 10g Distilled water up to 10g
Primjer 24 Example 24
Kobaltov hijaluronat kao aktivni sastojak 50 mg Cobalt hyaluronate as active ingredient 50 mg
Kalijev sorbat 10 mg Potassium sorbate 10 mg
Mekani, bijeli pčelinji vosak 125 mg Soft, white beeswax 125 mg
Sorbitan oleat 150 mg Sorbitan oleate 150 mg
Cetil stearil alkohol 840 mg Cetyl stearyl alcohol 840 mg
Gliceril monostearat 1100 mg Glyceryl monostearate 1100 mg
Propilen glikol 4750 mg Propylene glycol 4750 mg
Destilirana voda do 10 g Distilled water up to 10 g
Primjer 25 Example 25
Cinkov hijaluronat kao aktivni sastojak 50 mg Zinc hyaluronate as an active ingredient 50 mg
2-fenoksietanol 100 mg 2-phenoxyethanol 100 mg
Natrijev lauril sulfat 100 mg Sodium lauryl sulfate 100 mg
Cetil palmitat 400 mg Cetyl palmitate 400 mg
Stearin 400 mg Stearin 400 mg
Stearil alkohol 450 mg Stearyl alcohol 450 mg
Cetil alkohol 450 mg Cetyl alcohol 450 mg
Bijeli vazelin 500 mg White Vaseline 500 mg
Propilen glikol 550 mg Propylene glycol 550 mg
Glicerol 600 mg Glycerol 600 mg
Destilirana voda do 10,0 g Distilled water up to 10.0 g
Primjer 26 Example 26
Kobaltov hijaluronat kao aktivni sastojak 50 mg Cobalt hyaluronate as active ingredient 50 mg
2-fenoksietanol 100 mg 2-phenoxyethanol 100 mg
Natrijev lauril sulfat 100 mg Sodium lauryl sulfate 100 mg
Cetil palmitat 400 mg Cetyl palmitate 400 mg
Stearin 400 mg Stearin 400 mg
Stearil alkohol 450 mg Stearyl alcohol 450 mg
Cetil alkohol 450 mg Cetyl alcohol 450 mg
Bijeli vazelin 500 mg White Vaseline 500 mg
Propilen glikol 550 mg Propylene glycol 550 mg
Glicerol 600 mg Glycerol 600 mg
Destilirana voda do 10 g Distilled water up to 10 g
Primjer 27 Example 27
Cinkov hijaluronat kao aktivni sastojak 50 mg Zinc hyaluronate as an active ingredient 50 mg
Mikrokristalinički vosak 250 mg Microcrystalline wax 250 mg
Propilen glikol 500 mg Propylene glycol 500 mg
Sorbitol 400 mg Sorbitol 400 mg
Lanolin (acetiliran) 500 mg Lanolin (acetylated) 500 mg
Bijeli vazelin do 10 g White vaseline up to 10 g
V. Pripravci za čišćenje i cijeljenje gnojnih rana i opekotina V. Preparations for cleaning and healing purulent wounds and burns
Primjer 28 Example 28
Cinkov hijaluronat kao aktivni sastojak 10 mg Zinc hyaluronate as active ingredient 10 mg
Kalijev sorbat 1,0 mg Potassium sorbate 1.0 mg
Hidrofilni koloidni silicijev dioksid 50 mg Hydrophilic colloidal silicon dioxide 50 mg
Sorbitol do 1 g Sorbitol up to 1 g
Claims (15)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| HR930063 HRP930063B1 (en) | 1990-04-05 | 1993-02-01 | Novel compositions containing hyaluronic acid associates and a process for preparing the same |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| YU67190A YU47885B (en) | 1990-04-05 | 1990-04-05 | HYALURONIC ACID COMPLEXES, PROCEDURE FOR THEIR PREPARATION AND THEIR USE |
| HR930063 HRP930063B1 (en) | 1990-04-05 | 1993-02-01 | Novel compositions containing hyaluronic acid associates and a process for preparing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HRP930063A2 true HRP930063A2 (en) | 1995-10-31 |
| HRP930063B1 HRP930063B1 (en) | 2001-02-28 |
Family
ID=26316861
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HR930063 HRP930063B1 (en) | 1990-04-05 | 1993-02-01 | Novel compositions containing hyaluronic acid associates and a process for preparing the same |
Country Status (1)
| Country | Link |
|---|---|
| HR (1) | HRP930063B1 (en) |
-
1993
- 1993-02-01 HR HR930063 patent/HRP930063B1/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| HRP930063B1 (en) | 2001-02-28 |
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