US20080306022A1 - Agent for Applying to Mucosa and Method for the Production Thereof - Google Patents
Agent for Applying to Mucosa and Method for the Production Thereof Download PDFInfo
- Publication number
- US20080306022A1 US20080306022A1 US12/083,189 US8318906A US2008306022A1 US 20080306022 A1 US20080306022 A1 US 20080306022A1 US 8318906 A US8318906 A US 8318906A US 2008306022 A1 US2008306022 A1 US 2008306022A1
- Authority
- US
- United States
- Prior art keywords
- mucosa
- coo
- agent
- applying
- conh
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 210000004877 mucosa Anatomy 0.000 title claims abstract description 131
- 238000000034 method Methods 0.000 title claims description 47
- 238000004519 manufacturing process Methods 0.000 title description 6
- 229920002683 Glycosaminoglycan Polymers 0.000 claims abstract description 103
- 125000001165 hydrophobic group Chemical group 0.000 claims abstract description 85
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 49
- 210000005081 epithelial layer Anatomy 0.000 claims abstract description 18
- 239000004480 active ingredient Substances 0.000 claims abstract description 15
- 239000003795 chemical substances by application Substances 0.000 claims description 89
- -1 phenylethenyl group Chemical group 0.000 claims description 43
- 210000004087 cornea Anatomy 0.000 claims description 35
- 125000004432 carbon atom Chemical group C* 0.000 claims description 20
- 210000000214 mouth Anatomy 0.000 claims description 19
- 125000006850 spacer group Chemical group 0.000 claims description 15
- 229920002385 Sodium hyaluronate Polymers 0.000 claims description 14
- 229940010747 sodium hyaluronate Drugs 0.000 claims description 14
- 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 14
- 210000003932 urinary bladder Anatomy 0.000 claims description 14
- 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 claims description 12
- 125000005018 aryl alkenyl group Chemical group 0.000 claims description 12
- 125000003118 aryl group Chemical group 0.000 claims description 12
- 229920002674 hyaluronan Polymers 0.000 claims description 12
- 229960003160 hyaluronic acid Drugs 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 239000003889 eye drop Substances 0.000 claims description 9
- 229940012356 eye drops Drugs 0.000 claims description 9
- 125000000217 alkyl group Chemical group 0.000 claims description 8
- 125000003342 alkenyl group Chemical group 0.000 claims description 7
- 125000000304 alkynyl group Chemical group 0.000 claims description 7
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 7
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 6
- 125000005015 aryl alkynyl group Chemical group 0.000 claims description 6
- 230000002265 prevention Effects 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 125000000539 amino acid group Chemical group 0.000 claims description 5
- 210000000795 conjunctiva Anatomy 0.000 claims description 5
- 125000001072 heteroaryl group Chemical group 0.000 claims description 5
- 208000021921 corneal disease Diseases 0.000 claims description 4
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 claims description 3
- 229920002567 Chondroitin Polymers 0.000 claims description 3
- 229920001287 Chondroitin sulfate Polymers 0.000 claims description 3
- 229920000045 Dermatan sulfate Polymers 0.000 claims description 3
- 229920002971 Heparan sulfate Polymers 0.000 claims description 3
- 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 claims description 3
- 229920000288 Keratan sulfate Polymers 0.000 claims description 3
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 3
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 claims description 3
- 229940059329 chondroitin sulfate Drugs 0.000 claims description 3
- 229940051593 dermatan sulfate Drugs 0.000 claims description 3
- 229920000669 heparin Polymers 0.000 claims description 3
- 229960002897 heparin Drugs 0.000 claims description 3
- KXCLCNHUUKTANI-RBIYJLQWSA-N keratan Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@H](COS(O)(=O)=O)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@H](O[C@@H](O[C@H]3[C@H]([C@@H](COS(O)(=O)=O)O[C@@H](O)[C@@H]3O)O)[C@H](NC(C)=O)[C@H]2O)COS(O)(=O)=O)O[C@H](COS(O)(=O)=O)[C@@H]1O KXCLCNHUUKTANI-RBIYJLQWSA-N 0.000 claims description 3
- AVJBPWGFOQAPRH-FWMKGIEWSA-L dermatan sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS([O-])(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C([O-])=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-L 0.000 claims 2
- 125000000600 disaccharide group Chemical group 0.000 claims 2
- 208000035475 disorder Diseases 0.000 abstract description 42
- 230000003902 lesion Effects 0.000 abstract description 7
- 206010061218 Inflammation Diseases 0.000 abstract description 6
- 230000004054 inflammatory process Effects 0.000 abstract description 6
- 230000001747 exhibiting effect Effects 0.000 abstract description 2
- 230000001225 therapeutic effect Effects 0.000 abstract 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 84
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 84
- 229940114081 cinnamate Drugs 0.000 description 79
- 239000000126 substance Substances 0.000 description 79
- 210000001508 eye Anatomy 0.000 description 51
- 239000000243 solution Substances 0.000 description 51
- 241000283973 Oryctolagus cuniculus Species 0.000 description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 44
- 239000007864 aqueous solution Substances 0.000 description 42
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 36
- 230000035876 healing Effects 0.000 description 36
- 210000003560 epithelium corneal Anatomy 0.000 description 31
- 238000001704 evaporation Methods 0.000 description 29
- 230000008020 evaporation Effects 0.000 description 29
- 239000011780 sodium chloride Substances 0.000 description 28
- 150000001875 compounds Chemical class 0.000 description 25
- 230000002950 deficient Effects 0.000 description 21
- 230000000694 effects Effects 0.000 description 20
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 18
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 18
- 229960003299 ketamine Drugs 0.000 description 18
- BPICBUSOMSTKRF-UHFFFAOYSA-N xylazine Chemical compound CC1=CC=CC(C)=C1NC1=NCCCS1 BPICBUSOMSTKRF-UHFFFAOYSA-N 0.000 description 18
- 229960001600 xylazine Drugs 0.000 description 18
- NJDNXYGOVLYJHP-UHFFFAOYSA-L disodium;2-(3-oxido-6-oxoxanthen-9-yl)benzoate Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=CC(=O)C=C2OC2=CC([O-])=CC=C21 NJDNXYGOVLYJHP-UHFFFAOYSA-L 0.000 description 17
- 230000002209 hydrophobic effect Effects 0.000 description 17
- 238000005259 measurement Methods 0.000 description 16
- 239000002953 phosphate buffered saline Substances 0.000 description 16
- 239000007787 solid Substances 0.000 description 16
- 238000001356 surgical procedure Methods 0.000 description 16
- 238000011200 topical administration Methods 0.000 description 16
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 14
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 14
- 238000013508 migration Methods 0.000 description 14
- 230000005012 migration Effects 0.000 description 14
- 206010002091 Anaesthesia Diseases 0.000 description 13
- 230000037005 anaesthesia Effects 0.000 description 13
- 238000002360 preparation method Methods 0.000 description 13
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 12
- 206010013781 dry mouth Diseases 0.000 description 12
- 208000005946 Xerostomia Diseases 0.000 description 11
- 150000002016 disaccharides Chemical class 0.000 description 11
- 125000000524 functional group Chemical group 0.000 description 11
- 239000000203 mixture Substances 0.000 description 11
- 241000699800 Cricetinae Species 0.000 description 10
- 238000002347 injection Methods 0.000 description 10
- 239000007924 injection Substances 0.000 description 10
- 210000005252 bulbus oculi Anatomy 0.000 description 9
- 238000002695 general anesthesia Methods 0.000 description 9
- 238000002834 transmittance Methods 0.000 description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 238000010253 intravenous injection Methods 0.000 description 8
- 125000001273 sulfonato group Chemical group [O-]S(*)(=O)=O 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 7
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 7
- 238000001556 precipitation Methods 0.000 description 7
- 235000017557 sodium bicarbonate Nutrition 0.000 description 7
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 7
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 241000201841 Celosia Species 0.000 description 5
- 229940024606 amino acid Drugs 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 5
- 238000004132 cross linking Methods 0.000 description 5
- 239000006196 drop Substances 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 210000004400 mucous membrane Anatomy 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 230000000144 pharmacologic effect Effects 0.000 description 5
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000008215 water for injection Substances 0.000 description 5
- 0 *C(=O)[C@@H]1O[C@@H](OC)C(O)C(O)[C@@H]1O[C@@H]1OC(CO)[C@@H](O)[C@H](C)C1NC(C)=O Chemical compound *C(=O)[C@@H]1O[C@@H](OC)C(O)C(O)[C@@H]1O[C@@H]1OC(CO)[C@@H](O)[C@H](C)C1NC(C)=O 0.000 description 4
- 206010013774 Dry eye Diseases 0.000 description 4
- 206010037508 Punctate keratitis Diseases 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- IOQPZZOEVPZRBK-UHFFFAOYSA-N octan-1-amine Chemical compound CCCCCCCCN IOQPZZOEVPZRBK-UHFFFAOYSA-N 0.000 description 4
- 230000002085 persistent effect Effects 0.000 description 4
- 238000011907 photodimerization Methods 0.000 description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 3
- 125000004973 1-butenyl group Chemical group C(=CCC)* 0.000 description 3
- 125000004972 1-butynyl group Chemical group [H]C([H])([H])C([H])([H])C#C* 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 125000001931 aliphatic group Chemical group 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000003628 erosive effect Effects 0.000 description 3
- 210000000744 eyelid Anatomy 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000003020 moisturizing effect Effects 0.000 description 3
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- 210000000664 rectum Anatomy 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 210000003786 sclera Anatomy 0.000 description 3
- 210000001215 vagina Anatomy 0.000 description 3
- 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 class CC(=O)N[C@@H]1C[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O)[C@H](O)[C@H]1O WCDDVEOXEIYWFB-VXORFPGASA-N 0.000 description 2
- FJLUATLTXUNBOT-UHFFFAOYSA-N 1-Hexadecylamine Chemical compound CCCCCCCCCCCCCCCCN FJLUATLTXUNBOT-UHFFFAOYSA-N 0.000 description 2
- BMTZEAOGFDXDAD-UHFFFAOYSA-M 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium;chloride Chemical compound [Cl-].COC1=NC(OC)=NC([N+]2(C)CCOCC2)=N1 BMTZEAOGFDXDAD-UHFFFAOYSA-M 0.000 description 2
- GZAJOEGTZDUSKS-UHFFFAOYSA-N 5-aminofluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C21OC(=O)C1=CC(N)=CC=C21 GZAJOEGTZDUSKS-UHFFFAOYSA-N 0.000 description 2
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 208000003556 Dry Eye Syndromes Diseases 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 208000005615 Interstitial Cystitis Diseases 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 125000005196 alkyl carbonyloxy group Chemical group 0.000 description 2
- 208000002399 aphthous stomatitis Diseases 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- OSASVXMJTNOKOY-UHFFFAOYSA-N chlorobutanol Chemical compound CC(C)(O)C(Cl)(Cl)Cl OSASVXMJTNOKOY-UHFFFAOYSA-N 0.000 description 2
- 229930016911 cinnamic acid Natural products 0.000 description 2
- 235000013985 cinnamic acid Nutrition 0.000 description 2
- 201000005228 cornea cancer Diseases 0.000 description 2
- 208000024726 cornea neoplasm Diseases 0.000 description 2
- 201000003146 cystitis Diseases 0.000 description 2
- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 206010023332 keratitis Diseases 0.000 description 2
- 201000010666 keratoconjunctivitis Diseases 0.000 description 2
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 2
- 125000002950 monocyclic group Chemical group 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000001331 nose Anatomy 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- WGYKZJWCGVVSQN-UHFFFAOYSA-N propylamine Chemical group CCCN WGYKZJWCGVVSQN-UHFFFAOYSA-N 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 2
- OCUSNPIJIZCRSZ-ZTZWCFDHSA-N (2s)-2-amino-3-methylbutanoic acid;(2s)-2-amino-4-methylpentanoic acid;(2s,3s)-2-amino-3-methylpentanoic acid Chemical compound CC(C)[C@H](N)C(O)=O.CC[C@H](C)[C@H](N)C(O)=O.CC(C)C[C@H](N)C(O)=O OCUSNPIJIZCRSZ-ZTZWCFDHSA-N 0.000 description 1
- 125000006019 1-methyl-1-propenyl group Chemical group 0.000 description 1
- 125000006023 1-pentenyl group Chemical group 0.000 description 1
- 125000006017 1-propenyl group Chemical group 0.000 description 1
- 125000000069 2-butynyl group Chemical group [H]C([H])([H])C#CC([H])([H])* 0.000 description 1
- 125000006020 2-methyl-1-propenyl group Chemical group 0.000 description 1
- 125000000094 2-phenylethyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- 125000000474 3-butynyl group Chemical group [H]C#CC([H])([H])C([H])([H])* 0.000 description 1
- FNEHAOQZWPHONV-UHFFFAOYSA-N 9h-carbazole;sulfuric acid Chemical compound OS(O)(=O)=O.C1=CC=C2C3=CC=CC=C3NC2=C1 FNEHAOQZWPHONV-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 208000002177 Cataract Diseases 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 206010011793 Cystitis haemorrhagic Diseases 0.000 description 1
- 206010056246 Eosinophilic cystitis Diseases 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 208000005232 Glossitis Diseases 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241000699673 Mesocricetus auratus Species 0.000 description 1
- 206010028111 Mucosal dryness Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 206010034944 Photokeratitis Diseases 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 206010036774 Proctitis Diseases 0.000 description 1
- 206010036783 Proctitis ulcerative Diseases 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 201000002154 Pterygium Diseases 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229920001284 acidic polysaccharide Polymers 0.000 description 1
- 150000004805 acidic polysaccharides Chemical class 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- AVJBPWGFOQAPRH-FWMKGIEWSA-N alpha-L-IdopA-(1->3)-beta-D-GalpNAc4S Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS(O)(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C(O)=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-N 0.000 description 1
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000005428 anthryl group Chemical group [H]C1=C([H])C([H])=C2C([H])=C3C(*)=C([H])C([H])=C([H])C3=C([H])C2=C1[H] 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 239000008135 aqueous vehicle Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 229960000686 benzalkonium chloride Drugs 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 1
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 208000020670 canker sore Diseases 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 229960003333 chlorhexidine gluconate Drugs 0.000 description 1
- YZIYKJHYYHPJIB-UUPCJSQJSA-N chlorhexidine gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O.C1=CC(Cl)=CC=C1NC(=N)NC(=N)NCCCCCCNC(=N)NC(=N)NC1=CC=C(Cl)C=C1 YZIYKJHYYHPJIB-UUPCJSQJSA-N 0.000 description 1
- 229960004926 chlorobutanol Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 125000002541 furyl group Chemical group 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 210000005095 gastrointestinal system Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002070 germicidal effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 150000002337 glycosamines Chemical class 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 201000002802 hemorrhagic cystitis Diseases 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 229940014041 hyaluronate Drugs 0.000 description 1
- 125000001041 indolyl group Chemical group 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 206010023365 keratopathy Diseases 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 125000003136 n-heptyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001624 naphthyl group Chemical group 0.000 description 1
- 125000004998 naphthylethyl group Chemical group C1(=CC=CC2=CC=CC=C12)CC* 0.000 description 1
- 125000004923 naphthylmethyl group Chemical group C1(=CC=CC2=CC=CC=C12)C* 0.000 description 1
- 210000003928 nasal cavity Anatomy 0.000 description 1
- 210000002850 nasal mucosa Anatomy 0.000 description 1
- 229940105631 nembutal Drugs 0.000 description 1
- 125000001971 neopentyl group Chemical group [H]C([*])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 231100000989 no adverse effect Toxicity 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 125000002971 oxazolyl group Chemical group 0.000 description 1
- 125000004043 oxo group Chemical group O=* 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 239000002831 pharmacologic agent Substances 0.000 description 1
- 125000005561 phenanthryl group Chemical group 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 125000001844 prenyl group Chemical group [H]C([*])([H])C([H])=C(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- 125000000714 pyrimidinyl group Chemical group 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 125000000475 sulfinyl group Chemical group [*:2]S([*:1])=O 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Chemical group 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000001973 tert-pentyl group Chemical group [H]C([H])([H])C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 125000000335 thiazolyl group Chemical group 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7008—Compounds having an amino group directly attached to a carbon atom of the saccharide radical, e.g. D-galactosamine, ranimustine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
- A61K49/0043—Fluorescein, used in vivo
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0048—Eye, e.g. artificial tears
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/02—Nasal agents, e.g. decongestants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/10—Drugs for disorders of the urinary system of the bladder
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0034—Urogenital system, e.g. vagina, uterus, cervix, penis, scrotum, urethra, bladder; Personal lubricants
Definitions
- the present invention relates to an agent for applying to mucosa containing a hydrophobic group binding type glycosaminoglycan as an active ingredient, and a method for production thereof.
- GAG glycosaminoglycan
- photoreactive hyaluronic acid whose water-solubility has been increased by binding a photocrosslinking group such as cinnamic acid to hyaluronic acid and further giving an alkali treatment thereto has been known (e.g., Patent Document 2).
- This photoreactive hyaluronic acid has been provided by binding a photocrosslinking group such as cinnamic acid to hyaluronic acid in order to provide medical materials such as anti-adhesion materials by giving the photocrosslinking, and does not aim at enhancing the staying property in the mucosal tissue.
- Patent Document 1 Japanese Published Unexamined Patent Publication No. Hei-1-238530
- Patent Document 2 Japanese Published Unexamined Patent Publication No. 2002-249501
- the present invention aims at providing an agent for applying to mucosa, which exerts an excellent staying property and pharmacological effects in mucosa.
- hydrophobic group binding type GAG obtained by binding a hydrophobic group to GAG via a binding chain can be used as an extremely excellent active ingredient in an agent for applying to mucosa because this GAG keeps healing effects inherent in GAG on mucosal disorders such as inflammation and lesions and exhibits a high staying property when applied thereto, and have completed the present invention.
- the present invention relates to an agent for applying to mucosa which contains glycosaminoglycan (GAG) into which the hydrophobic group is introduced via the binding chain.
- GAG glycosaminoglycan
- the agent for applying to mucosa of the present invention can exert the persistent healing effect on the mucosal disorder such as inflammation and lesions even by low frequent administration because this can stay at a diseased site for a long period of time by exhibiting high staying property in the mucosa.
- FIG. 1 is a view showing a spectrum of light transmittance
- FIG. 2 is a view showing healed area percentages
- FIG. 3 is a view showing healed areas
- FIG. 4 is a view showing healing rate
- FIG. 5 is a view showing healed areas
- FIG. 6 is a view showing healing rate
- FIG. 7 is a view showing healed areas
- FIG. 8 is a view showing healing rate
- FIG. 9 is a view showing healed areas
- FIG. 10 is a view showing healing rate
- FIG. 11 is a view showing staying property at peeled sites in rabbit corneal epithelia
- FIG. 12 is a view showing staying property at peeled sites in rabbit corneal epithelia
- FIG. 13 is a view showing photographs of an eyeball after irradiation with ultraviolet rays
- FIG. 14 is a view showing amounts of water evaporation in a removed cornea
- FIG. 15 is a view showing amounts of water evaporation in a removed cornea.
- FIG. 16 is a view showing changes in water evaporation amount ratio in model hamsters for xerostomia.
- FIG. 17 is a view showing healed areas
- FIG. 18 is a view showing healing rate
- FIG. 19 is a view showing healed areas
- FIG. 20 is a view showing healing rate.
- an alkyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms.
- An alkenyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms, having at least one double bond.
- An alkynyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms, having at least one triple bond.
- An aryl group refers to a monocyclic or polycyclic aromatic hydrocarbon group having 6 to 20 carbon atoms as ring-constituting atoms.
- a heteroaryl group refers to a monocyclic or polycyclic aromatic hydrocarbon group having 3 to 20 carbon atoms and one or more heteroatoms selected from nitrogen, sulfur and oxygen atoms as the ring-constituting atoms.
- An arylalkyl group refers to the alkyl group defined above substituted with the aryl group defined above.
- An arylalkenyl group refers to the alkenyl group defined above substituted with the aryl defined above.
- An arylalkynyl group is the alkynyl group defined above substituted with the aryl group defined above.
- An amino acid group refers to a group derived by losing a carboxyl group, an amino group or a hydroxyl group by a chemical bond from a natural or synthetic amino acid.
- the term “treatment” includes prevention, control of progression (prevention of deterioration), improvement (reduction) and cure of the mucosal disorder.
- the “mucosal disorder” means a condition where morphology, properties and functions to be inherent in the mucosa are disordered in some form.
- the mucosal disorder can include the conditions such as lesions, defects, erosion, inflammation, ulcers and dryness.
- GAG into which the hydrophobic group is introduced via the binding chain, which is contained as the active ingredient in the agent for applying to mucosa of the present invention can be any GAG as long as the GAG binds the group having hydrophobicity derived from a hydrophobic compound having a water insoluble and oil soluble nature. This hydrophobic group is bound to GAG via the binding chain. As described later, it is not necessary that all constitutive units of GAG bind the hydrophobic groups.
- GAG in GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention is an acidic polysaccharide having a repeated long chain structure of disaccharide composed of amino sugar and uronic acid (or galactose).
- examples of such GAG include hyaluronic acid, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan sulfate and keratan sulfate, and among them, hyaluronic acid is preferable.
- These GAG may be pharmaceutically acceptable salts thereof.
- GAG in the agent for applying to mucosa of the present invention is sodium hyaluronate.
- An origin of GAG is not particularly limited, and GAG may be derived from an animal or a microorganism or chemically synthesized.
- the molecular weight of GAG is not particularly limited, but its weight average molecular weight is preferably 200,000 to 3,000,000, more preferably 500,000 to 2,000,000 and most preferably 600,000 to 1,200,000.
- hyaluronic acid or the pharmaceutically acceptable salt thereof is used, its weight average molecular weight is preferably 200,000 to 3,000,000, more preferably 500,000 to 2,000,000 and most preferably 600,000 to 1,200,000.
- the hydrophobic group in GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention is any group as long as the hydrophobic group is derived from the compound having the water insoluble and oil soluble nature.
- Examples of such a group can include alkyl groups having 2 to 18 carbon atoms, alkenyl groups having 2 to 18 carbon atoms, alkynyl groups having 2 to 18 carbon atoms, aryl groups, heteroaryl groups, arylalkyl groups, arylalkenyl groups, arylalkynyl groups and amino acid groups.
- the alkyl groups having 2 to 18 carbon atoms can include methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-butyl, n-pentyl, t-pentyl, isopentyl, neopentyl, n-heptyl, 5-methylhexyl, 4,4-dimethyl-pentyl, 1,1-dimethyl-pentyl and n-octyl.
- the alkyl groups such as n-butyl having 2 to 6 carbon atoms such as n-butyl can be preferably included.
- the alkenyl groups having 2 to 18 carbon atoms can include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-methyl-1-propenyl, 1-methyl-1-propenyl, 1-pentenyl, 3-methyl-2-butenyl, 1-heptene-1-yl and 2-heptene-1-yl.
- the alkenyl groups such as 1-butenyl having 2 to 6 carbon atoms such as 1-butenyl can be preferably included.
- the alkynyl groups having 2 to 18 carbon atoms can include ethynyl, 1-propinyl, 2-propinyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-heptynyl, 2-heptynyl and 3-heptynyl.
- the alkynyl groups such as 1-butynyl having 2 to 6 carbon atoms such as 1-butynyl can be preferably included.
- the aryl groups can include groups such as phenyl, naphthyl, anthryl and phenanthryl.
- the heteroaryl groups can include groups such as furyl, thionyl, thiazolyl, oxazolyl, pyrrolyl, pyridyl, pyrimidinyl and indolyl.
- the arylalkyl groups can include groups such as benzyl, phenethyl, naphthylmethyl and naphthylethyl.
- the arylalkenyl groups can include groups such as 2-phenyl-ethenyl and p-aminophenylethenyl.
- the arylalkynyl groups can include groups such as 2-phenyl-ethynyl and p-aminophenylethynyl.
- the amino acid groups can include groups derived from aliphatic amino acids such as glycine, alanine and ⁇ -alanine; branched aliphatic amino acids such as leucine, isoleucine and valine; aromatic amino acids such as phenylalanine and tyrosine; and heterocyclic amino acids such as tryptophan and histidine.
- aliphatic amino acids such as glycine, alanine and ⁇ -alanine
- branched aliphatic amino acids such as leucine, isoleucine and valine
- aromatic amino acids such as phenylalanine and tyrosine
- heterocyclic amino acids such as tryptophan and histidine.
- hydrophobic groups may be monosubstituted or polysubstituted with groups such as hydroxyl, carboxyl, cyano, amino (which may be monosubstituted or disubstituted with the above alkyl), nitro, oxo and alkylcarbonyloxy.
- aryl groups, arylalkyl groups, arylalkenyl groups and arylalkynyl groups which are the hydrophobic groups containing the aryl group can be preferably included, and the arylalkenyl group and the aryl group substituted with the alkylcarbonyloxy group can be particularly preferably included.
- an arylalkenyl group it is possible to specifically use phenylethenyl and p-aminophenylethenyl.
- the aryl group it is possible to preferably use the groups such as CH 3 —(CH 2 ) l —COO-phenyl (wherein l represents 0 or an integer of 1 to 18).
- hydrophobic groups may also have a function such as ultraviolet ray absorption ability due to having a double bond in the hydrophobic group as shown by a functional group such as phenyl-ethenyl exemplified above contained in the hydrophobic group.
- a functional group such as phenyl-ethenyl exemplified above contained in the hydrophobic group.
- the agent for applying to mucosa of the present invention when used for the treatment of corneal epithelial layer disorders such as corneal xerosis (dry eye), keratoconjunctivitis, superficial punctate keratitis (SPK), corneal epithelial erosion, corneal epithelial loss and corneal tumor, it is possible to make the agent for applying to mucosa which has pharmacological effects on the above disorders in combination with the function effectively absorbing the harmful ultraviolet rays by the use of the group having the ultraviolet ray absorption ability as the hydrophobic group.
- corneal epithelial layer disorders such as corneal xerosis (dry eye), keratoconjunctivitis, superficial punctate keratitis (SPK), corneal epithelial erosion, corneal epithelial loss and corneal tumor
- the group having the ultraviolet ray absorption ability for example, arylalkenyl group having the conjugation double bond which is exemplified by 2-phenyl-ethenyl and p-aminophenylethenyl described above is preferable.
- “GAG into which the hydrophobic group is introduced” which is the active ingredient of the agent for applying to mucosa of the present invention is made into an aqueous solution of 0.1% by weight, which blocks 70 to 100% transmission of the ultraviolet rays at a wavelength of 200 to 300 nm when an ultraviolet ray transmittance is measured by the method described in the Example described later.
- Such a hydrophobic group having the ultraviolet ray absorption ability can preferably include the arylalkenyl groups such as 2-phenyl-ethenyl and p-aminophenylethenyl.
- GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention, the above GAG is bound to the above hydrophobic group via the binding chain.
- GAG has the functional group which is a carboxyl, hydroxyl or sulfonate (—SO 3 H) group as the side chain.
- the hydrophobic group can be bound to GAG via the binding chain obtained by forming an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with these functional groups.
- Such a binding chain can specifically include —CONH—, —COO—, —O—, —SO 3 — and —SO 2 NH—.
- the carboxylic acid amide bond of —CONH— and the carboxylate ester bond of —COO— can be preferably used, and the carboxylic acid amide bond of —CONH— can be particularly preferably used.
- the hydrophobic group is bound to GAG via the above binding chain, and a spacer chain may further exist between the binding chain and the hydrophobic group.
- a spacer chain any chain group can be used as long as the spacer group does not completely lose the pharmacological effects which GAG has.
- —(CH 2 ) m — and —(CH 2 )—(OCH 2 ) n — (wherein m and n are integers of 1 to 18, respectively) can be included.
- spacer chains can further have the binding chains such as —CONH—, —COO—, —O—, —SO 3 — and —SO 2 NH— which are the same as above at the hydrophobic group side.
- Such a spacer chain having the binding chain at the hydrophobic group side can specifically include —COO—(CH 2 ) m —, —COO—(CH 2 )—(OCH 2 ) n —, —CONH—(CH 2 ) m — and —CONH—(CH 2 )—(OCH 2 ) n —.
- GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention it is not necessary that all of GAG constitutive units respectively have the hydrophobic groups.
- a ratio of the bound hydrophobic group in molar equivalent relative to a disaccharide repeat unit in molar equivalent of GAG (hereinafter, referred to as an “introduction ratio”) can be optionally determined depending on the type of the hydrophobic group, the degree of required hydrophobicity, the type of the mucosal disorder administered with the agent for applying to mucosa and the administration site, etc.
- a phenylethenyl group which may be substituted as the hydrophobic group preferably 5 to 30% and more preferably 10 to 20% of a hydrophobic group in molar equivalent is introduced relative to the disaccharide repeat unit in molar equivalent of GAG (in the case where the crosslinking bond described later is not formed).
- the hydrophobic group may form a crosslinking bond between GAG molecules by the functional group contained in the group.
- the hydrophobic group capable of forming the crosslinking bond any group can be used as long as the hydrophobic group produces a photodimerization reaction or a photopolymerization reaction by irradiation of ultraviolet rays and is the same as defined above.
- the hydrophobic group capable of forming the crosslinking bond includes, for example, phenylethenyl, p-aminophenylethenyl, ethenyl, 2-carboxyethenyl and pentane-1,3-dienyl. It is desirable that these groups are bound to GAG via the binding chain which contains the carbonyl group.
- phenylethenyl or p-aminophenylethenyl which is bound to GAG via the binding chain which contains the carbonyl group can be particularly preferably used.
- GAG molecules can be crosslinked with one another by being subjected to the photodimerization reaction or the photopolymerization reaction by standard methods.
- the photodimerization reaction or the photopolymerization reaction can be given.
- GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention can specifically include the agents for applying to mucosa containing GAG into which Ph-CH ⁇ CH—COO—(CH 2 ) m —NHCO—; Ph-CH ⁇ CH—COO—CH 2 —(OCH 2 ) n —NHCO—; Ph-CH ⁇ CH—CONH—(CH 2 ) m —NHCO—; Ph-CH ⁇ CH—CONH—CH 2 —(OCH 2 ) n —NHCO—; Ph-CH ⁇ CH—COO—(CH 2 ) m —O—CO—; Ph-CH ⁇ CH—COO—CH 2 —(OCH 2 ) n —O—CO—; Ph-CH ⁇ CH—CONH—(CH 2 ) m —O—CO—; Ph-CH ⁇ CH—CONH—(CH 2 ) m —O—CO—; Ph-CH ⁇ CH—CONH—(CH
- GAG having the repeat unit of the structural unit represented by the Chemical formula 1, as a basic skeleton:
- R represents R 1 or R 2 ; Ac represents an acetyl group; R 1 represents ONa or OH; R 2 represents (1) Ph-CH ⁇ CH—COO—(CH 2 ) m —NH—; (2) Ph-CH ⁇ CH—COO—CH 2 —(OCH 2 ) n —NH—; (3) Ph-CH ⁇ CH—CONH—(CH 2 ) m —NH; (4) Ph-CH ⁇ CH—CONH—CH 2 —(OCH 2 ) n —NH—; (5) Ph-CH ⁇ CH—COO—(CH 2 ) m —O—; (6) Ph-CH ⁇ CH—COO—CH 2 —(OCH 2 ) n —O—; (7) Ph-CH ⁇ CH—CONH—(CH 2 ) m —O—; (8) Ph-CH ⁇ CH—CONH—CH 2 —(OCH 2 ) n —O—; (9) CH 3 —(CH 2 ) l —COO-Ph-
- GAG into which the hydrophobic group is introduced via the binding chain
- GAG is reacted with a hydrophobic compound in which the above hydrophobic group has been bound to the functional group such as hydroxyl, carboxyl, amino or sulfonate group which can form an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the carboxyl, hydroxyl or sulfonate (—SO 3 H) group in GAG.
- a hydrophobic compound in which the above hydrophobic group has been bound to the functional group such as hydroxyl, carboxyl, amino or sulfonate group which can form an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the carboxyl, hydroxyl or sulfonate (—SO 3 H) group in GAG.
- GAG having the carboxyl group is reacted with the hydrophobic compound having an amino group to bind the carboxyl group in GAG to the amino group in the hydrophobic compound.
- GAG is reacted with the hydrophobic compound having hydroxyl or carboxyl group to bind the carboxyl group in GAG to the hydroxyl group in the hydrophobic compound or bind the hydroxyl group in GAG to the carboxyl group in the hydrophobic compound.
- GAG having the hydroxyl group is reacted with the hydrophobic compound having the hydroxyl group to react the hydroxyl group in GAG with the hydroxyl group in the hydrophobic compound.
- GAG is reacted with the hydrophobic compound having the hydroxyl group or sulfonate group to bind the hydroxyl group in GAG to the sulfonate group in the hydrophobic compound or bind the sulfonate group in GAG to the hydroxyl group in the hydrophobic compound.
- the order in introducing the spacer chain and the hydrophobic group to GAG is not particularly limited.
- a spacer compound having the functional group such as the hydroxyl, carboxyl, amino or sulfonate group which can form the ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the functional group in GAG at one end of the above spacer chain is reacted with GAG, and subsequently, the other end of the spacer compound is reacted with the hydrophobic compound which is bound to the functional group such as the hydroxyl, carboxyl, amino or sulfonate group, or the method in which the spacer compound having the functional group such as the hydroxyl, carboxyl, amino or sulfonate group which can form an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid
- a condensing agent can preferably include water soluble carbodiimide such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI.HCl), condensing agents such as dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinate imide (HOSu).
- a condensing agent can preferably include water soluble carbodiimide such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI.HCl), condensing agents such as dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinate imide (HOSu).
- the condensation method using water soluble carbodiimide such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) and N-hydroxysuccinate imide can be preferably used.
- the reaction can be accomplished using a mixed solvent of water and a water soluble organic solvent such as dioxane, dimethylformamide or ethanol.
- a water soluble organic solvent such as dioxane, dimethylformamide or ethanol.
- the hyaluronate derivative which is highly soluble in an aqueous vehicle can be obtained by treating with a base such as sodium hydrogen carbonate after the completion of the reaction.
- the crosslink can be formed by irradiating light to the solution containing them using an ultraviolet lamp.
- the agent for applying to mucosa of the present invention contains one or more GAG into which the hydrophobic group is introduced via the binding chain as the active ingredient, and may also further include other medically, pharmaceutically or biologically acceptable substances other than the GAG into which the hydrophobic group is introduced via the binding chain.
- Such substances include but are not limited to, salts such as sodium chloride, potassium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate and monopotassium hydrogen phosphate, and preservatives such as paraoxybenzoate esters, benzalkonium chloride, chlorobutanol and chlorhexidine gluconate, and other pharmacologically active ingredients.
- the agent for applying to mucosa of the present invention can be made into any publicly known formulation forms (e.g., solid preparations such as granules and powder, liquid preparations such as aqueous solutions, suspension and emulsion, and gel preparations) as the pharmaceutical for applying to the mucosa.
- the form thereof upon formulating and distributing and the form thereof upon applying to the mucosa may be the same or different.
- the agent for applying to mucosa of the present invention may be formulated in the form of solution and may be applied directly to the mucosa as it is.
- the agent for applying to mucosa of the present invention may be formulated and distributed in the solid form, and may be made into solution or gel when being applied to the mucosa.
- the agent for applying to mucosa of the present invention can be made into the formulation form for being prepared when used.
- the amount of GAG into which the hydrophobic group is introduced via the binding chain is preferably 0.02 to 5% by weight, more preferably 0.1 to 3% by weight, extremely preferably 0.1 to 1% by weight and most preferably 0.1 to 0.6% by weight.
- the agent for applying to mucosa of the present invention aims at applying to the mucosa.
- Animals to which the agent for applying to mucosa of the present invention is applied are not particularly limited as long as they have the mucosa, and mammalian animals are preferable.
- the mammalian animals include, but are not limited to, humans, horses, cattle, dogs, cats, rabbits, hamsters, guinea pigs and mice.
- the agent for applying to mucosa of the present invention may be of course made into the pharmaceuticals for humans, and can also be made into the pharmaceuticals for the animals. Among them, it is preferable to be made into the pharmaceuticals for humans.
- the mucosa to which the agent for applying to mucosa of the present invention can be applied is not particularly limited as long as the mucosa is the mucosa present in the animal.
- Such mucosa include mucosal tissues present in organs and tissues exemplified by the gastrointestinal system such as the stomach and intestines, the cardiovascular system, the respiratory system, the excretion system such as the urinary bladder, rectum and anus, the genital system such as the vagina, and organs such as eyes, nose and oral cavity which contact with the external world.
- the agent for applying to mucosa of the present invention can be preferably applied to the cornea, conjunctiva, oral cavity mucosa and urinary bladder mucosa.
- the agent for applying to mucosa of the present invention can be widely applied to such mucosa.
- the purpose of the application is not particularly limited, and for example, the purposes such as protection of the mucosal tissue (e.g., prevention of snow blindness by ultraviolet rays, pterygium and cataract), prevention of mucosal dryness and the treatment of mucosal disorder can be exemplified.
- the agent for applying to mucosa of the present invention can be applied to not only the mucosa in the abnormal state (e.g., mucosa where the disorder has occurred) but also the mucosa in the normal state.
- the agent for applying to mucosa of the present invention exerts excellent pharmacological effects in the mucosa where the disorder has occurred, it is possible to preferably use for the treatment of the mucosal disorders, e.g., disorders in the cornea, conjunctiva, oral cavity mucosa and urinary bladder mucosa.
- the mucosal disorders e.g., disorders in the cornea, conjunctiva, oral cavity mucosa and urinary bladder mucosa.
- the agent for applying to mucosa of the present invention exerts excellent pharmacological effects particularly on disorders in the mucosal epithelia among the mucosal disorders, it is possible to be preferably used for the treatment of the disorders in the mucosal epithelia.
- disorders in the mucosal epithelia include corneal epithelial layer disorders such as corneal xerosis (dry eye); keratoconjunctivitis, superficial punctate keratitis (SPK), corneal epithelial erosion, corneal epithelial loss and corneal tumor; oral cavity mucosal disorders such as xerostomia (dry mouth), aphthous ulcer, stomatitis and glossitis; dryness and pruritus of nasal mucosa; urinary bladder mucosal disorders such as interstitial cystitis; ulcerative proctitis, and dryness of the rectum or vagina. Also dryness and lesions of organ mucosa upon surgical operation can be exemplified. Among them, it is possible to be preferably used for the treatment of the corneal epithelial layer disorders, the oral cavity mucosal epithelial disorders and the urinary bladder mucosa epithelial layer disorders.
- corneal epithelial layer disorders such as corneal
- the agent for applying to mucosa of the present invention can be applied to the mucosal tissues exemplified above, and its application method and application formulation can be appropriately determined by those skilled in the art depending on the position, morphology, property and function of the mucosa to be applied, and the purpose of the application.
- the agent for applying to mucosa of the present invention is applied to the mucosa in the liquid form such as solution in use.
- the liquid upon producing (formulating) or applying the agent for applying to mucosa of the present invention, the liquid can be obtained by dissolving GAG into which the hydrophobic group is introduced via the binding chain in the solvent.
- the solvent is not particularly limited as long as the solvent can dissolve the GAG into which the hydrophobic group is introduced via the binding chain and is the pharmaceutically acceptable solvent.
- a buffer such as a phosphate buffer or saline can be used, but the solvent is not limited thereto.
- the concentration of the GAG into which the hydrophobic group is introduced via the binding chain in the liquid agent is not particularly limited, and can be appropriately determined depending on the type of the mucosa to be applied and the degree of the mucosal disorder.
- the concentration is preferably 0.02 to 5% by weight, more preferably 0.1 to 3% by weight, still more preferably 0.1 to 1% by weight, still more preferably 0.1 to 0.6% by weight, extremely preferably 0.1 to 0.5% by weight and most preferably 0.1 to 0.3% by weight.
- an oral administration or the administration using a catheter can be selected.
- the administration method such as instillation of drop, nasal instillation or oral inclusion can be selected.
- the method of administering by injecting, spraying or applying the agent for applying to mucosa of the present invention to a lumen or a surface of these organs or tissues can be selected, but the methods are not limited thereto.
- the amount, the number of times and the frequency of the application (administration of) of the agent for applying to mucosa of the present invention is not particularly limited, and should be determined depending on the mucosa subjected to the application, the purpose of the application, the type, age, body weight, gender, and degree of mucosal disorder in the animal to be applied.
- the agent for applying to mucosa of the present invention when used for the purpose of treating the human corneal epithelial layer disorder, the agent for applying to'mucosa of the present invention at the above-described concentration as the liquid formulation for the instillation of drops (eye drops) containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by instilling 1 to 3 drops per administration 1 to 5 times per day, and may be administered by instilling 1 to 3 drops per administration 1 to 3 times per day.
- the agent for applying to mucosa of the present invention when used for the purpose of treating the human oral cavity mucosal disorder, the agent for applying to mucosa of the present invention at the above-described concentration as the liquid containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by putting the agent for applying to mucosa of the present invention in the oral cavity 1 to 5 times per day and rinsing for approximately several tens of seconds (preferably approximately 20 to 30 seconds) followed by spitting it out.
- the agent for applying to mucosa of the present invention is applied to the urinary bladder mucosal disorder
- this is preferably used for the treatment of the urinary bladder mucosal disorders exemplified by non-bacterial refractory cystitis exemplified by interstitial cystitis, eosinophilic cystitis and hemorrhagic cystitis which do not respond to anti-bacterial agents although symptoms similar to those of acute bacterial cystitis are exhibited.
- the agent for applying to mucosa of the present invention at the above-described concentration as the liquid containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by administering the agent for applying to mucosa of the present invention directly to the urinary bladder at the amount of 50 mL per administration 1 to 7 times per week or administering with a catheter in the urinary bladder.
- the agent for applying to mucosa of the present invention can stay at the diseased site for a longer period of time because the active ingredient contained in the agent exhibits the high staying property in the mucosa, compared with the conventional drugs containing hyaluronic acid as the active ingredient in which no hydrophobic group has been bound. Therefore, the agent for applying to mucosa of the present invention can also exert the treating effect persistently even at the low administration frequency on the disorders such as inflammation and lesions in the mucosa.
- the agent for applying to mucosa of the present invention is not limited by its administration frequency.
- sodium hyaluronate (1.06 g, 2.7° mmol/disacc
- the introduction ratio of the cinnamate derivative was 16%.
- the introduction ratio of the cinnamate derivative was calculated based on the amount of cinnamate by an absorbance measurement method (wavelength: 269 nm) and the amount of hyaluronate by a carbazole sulfate method.
- a 75 mg/5 mL aqueous solution of HOSu, a 62 mg/5 mL WFI solution of EDCI.HCl, and a 92 mg/5 mL WFI solution of 6-aminohexyl cinnamate hydrochloride (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.0 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 1,500,000; derived from cock's comb, Seikagaku Corporation) in water for injection (hereinafter referred to as WFI) (150 mL)/dioxane (75 mL).
- WFI sodium hyaluronate
- cinnamate derivative-introduced HA (12.5 g) was dissolved in phosphate buffered saline (concentration of phosphate: 1.5 mM, hereinafter abbreviated as “PBS”) to prepare 2.5% solution of cinnamate derivative-introduced HA (500 mL).
- PBS phosphate buffered saline
- the 2.5% solution of cinnamate derivative-introduced HA was irradiated by 800 W high pressure mercury lamp and performed by a heat treatment in an autoclave at 121° C. for 7.5 min. to yield crosslinked cinnamate derivative-introduced HA.
- a 172 mg/5 mL aqueous solution of HOSu, a 143 mg/5 mL aqueous solution of EDCI.HCl, and a 181 mg/5 mL aqueous solution of 3-aminopropyl cinnamate hydrochloride (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.0 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 900,000; derived from cock's comb, Seikagaku Corporation) in water (150 mL)/dioxane (75 mL).
- a 3.0 mmol/mL aqueous solution of HOSu, a 1.5 mmol/mL aqueous solution of EDCI.HCl and a 1.5 mmol/mL aqueous solution of 4-aminofluorescein (Tokyo Chemical Industry Co., Ltd.) were added to a solution of cinnamate derivative-introduced HA obtained in the above (3-1) (1.00 g, 2.5 mmol/disaccharide unit) in water (150 mL)/dioxane (75 mL). The mixture was stirred one day and a 500 mg/10 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added.
- a 3.0 mmol/mL aqueous solution of HOSu, a 1.5 mmol/mL aqueous solution of EDCI.HCl and a 1.5 mmol/mL aqueous solution of 4-aminofluorescein (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.00 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 900,000; derived from cock's comb, Seikagaku Corporation) in water (150 mL)/dioxane (75 mL).
- the 0.1% by weight aqueous solution of cinnamate derivative-introduced HA obtained in the above (1-1) was prepared, and the ultraviolet ray transmittance was measured by an spectrometer (UV-1600, Shimadzu Corporation).
- FIG. 1 A spectrum which indicates the transmittance is shown in FIG. 1 , and the transmittance (%) at various wavelengths is shown in Table 1. As a result, 100% of the transmittance was shown at the wavelengths of 340 nm or more, but the transmittance at the wavelengths of approximately 320 nm or less was 20% or less which was extremely low, and it was demonstrated that this solution effectively blocks the transmission of the ultraviolet ray.
- FIG. 1 scales are shown with 65 nm intervals on a horizontal axis and with 20% intervals on a vertical axis.
- ⁇ T (%) 234 19.9 340 106.2 380 101.8 450 98.5
- ⁇ and T represent the wavelength and the transmittance (%), respectively.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the final administration at 3 days after the peeling. When photographed, a focal length was made constant to make a magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using an image analyzer. A value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after peeling the corneal epithelia was performed was rendered as “healed area.”
- the results of a healed area percentage in each individual are shown in FIG. 2 , and the results of the healed area percentage and a healed area percentage ratio in each individual are shown in Table 2.
- the healed area percentage and the healed area percentage ratio were calculated as follows.
- Healed area percentage (%) (Healed area/Peeled area) ⁇ 100
- Healed area percentage ratio (Healed area percentage in right eye/Healed area percentage in left eye) ⁇ 100
- the left column shows the healed area percentage in the right eye (administration of cinnamate derivative-introduced HA), and the right column shows the healed area percentage in the left eye (administration of saline).
- the apparent effect to facilitate the healing of the corneal epithelial layer disorder was observed in 5 of the administered 6 individuals.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer.
- the value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.”
- the results of the healed area in each individual are shown in FIG. 3
- the results of a healing rate in each individual are shown in FIG. 4 .
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling* ⁇ Area at each time point (after 1 to 3 days)
- area after peeling means ‘just before the administration of the subject substance one hour after the cornea was peeled’ in calculation of healed area.
- FIGS. 3 and 4 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.5% by weight cinnamate derivative-introduced HA solution, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.5% by weight cinnamate derivative-introduced HA solution.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer.
- the value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.”
- the results of the healed area in each individual are shown in FIG. 5
- the results of a healing rate in each individual are shown in FIG. 6 .
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling ⁇ Area at each time point (after 1 to 3 days)
- FIGS. 5 and 6 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.3% by weight cinnamate derivative-introduced HA solution at all time points of days 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.3% by weight cinnamate derivative-introduced HA solution.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer.
- the value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.”
- the results of the healed area in each individual are shown in FIG. 7
- the results of the healed area and the healing rate in each individual are shown in FIG. 8 .
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling ⁇ Area at each time point (after 1 to 3 days)
- FIGS. 7 and 8 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution, compared with the healed area of the corneal epithelia in the control eyes administered with the 0.1% by weight HA aqueous solution. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 6 hours after the administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled site 3 hours after the final administration at, 3′ days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as the “healed area.”
- the results of the healed area in each individual are shown in FIG. 9
- the results of the healing rate in each individual are shown in FIG. 10 .
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling ⁇ Area at each time point (after 1 to 3 days)
- FIGS. 9 and 10 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution at all time points of the days 1 to 3, compared with the healed area of the corneal epithelia in the control eyes administered with the 0.1% by weight HA aqueous solution. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- Two rabbits were given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine per rabbit, and eyeballs were removed 30 minutes, one hour, one and a half hours and 2 hours and 30 minutes after the administration of the subject substance and the control substance.
- a pore was opened between the cornea and sclera in the removed eyeball using a surgical knife, and only the cornea was taken out using the microscissors.
- the removed cornea was placed on a biological sample slicing plate (supplied from Nisshin EM Corporation, Cat No.
- the frozen block was removed from the cryostat tray, and attached on a sample table using the O.C.T. compound.
- the sample table and a disposable microtome blade (supplied by Leica Microsystems Japan, Model 818, Lot. No. 913212) were set in a high performance frozen microtome for research, and the block was sliced under the condition of a frozen chamber temperature (CT) at ⁇ 20° C. and sample side temperature (OT) at ⁇ 16° C. to make sections with a thickness of 5 ⁇ m using silane coating slide glasses (supplied by Muto Pure Chemicals Co., Ltd., Star Frost Slide Glass, Cat. No. 5116).
- CT frozen chamber temperature
- OT sample side temperature
- the frozen section was set in an incident-light fluorescence microscope (Olympus Corporation, BH2-RFC), FA images and autofluorescent images were observed at IB cube (BH2-DMIB, excitation wavelength: 495 nm, absorption wavelength: 460 nm) and U cube (BH2-DMU, broad band U excitation, absorption wavelength: 435 nm), respectively.
- the FA image and the autofluorescent image were photographed using a cooled high sensitivity CCD camera (Keyence Corporation, VB-6010) under the condition of exposure time for one second and ISO sensitivity of 200.
- Photographs of the sampled cornea were shown in FIGS. 11 and 12 . From FIGS. 11 and 12 , it was identified by color development of the fluorescence label that the aqueous solution of 0.3% by weight fluorescence labeled HA which was the control substance stayed until 30 minutes after the administration but did not stay after one hour. Meanwhile, although the fluorescent color development of the aqueous solution of 0.3% by weight fluorescence labeled cinnamate derivative-introduced HA which was the subject substance was weakened with the elapse of time, the color development was observed at all time points from 30 minutes to 2 hours and 30 minutes after the administration, thereby the high staying performance was confirmed.
- Ultraviolet rays were irradiated to both eyes from a distance of approximately 10 cm apart from the rabbit eyeball using a 15 kW germicidal lamp. The irradiation was performed for 3 hours.
- the eyeball was stained with 0.2% sodium fluorescein under the continuous anesthesia of the rabbit, and photographed under violet light. When photographed, the focal length was made constant to make the magnification of photographs constant.
- Photographs after the irradiation of the ultraviolet rays were shown in FIG. 13 . From FIG. 13 , in the eyeball irradiated with the ultraviolet rays after the administration of the control substance, the disordered site stained with 0.2% sodium fluorescein was apparent. Meanwhile, in the eyeball irradiated with the ultraviolet rays after the administration of the subject substance, the disordered site stained with 0.2% sodium fluorescein was clearly smaller than that of the control substance, and the corneal disorder caused by the ultraviolet ray was prevented.
- Example 1 Using the removed cornea of the rabbit, the moisturizing performance of the cinnamate derivative-introduced HA prepared in Example 1 was validated.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, and the eyeball was removed. A pore was opened between the cornea and the sclera in the removed-eyeball using a surgical knife, and only the cornea was taken out using the microscissors.
- the drying treatment was performed by placing the removed cornea on a paraffin block with a False-tooth stability material and providing cold air from the distance of approximately 1 m from the cornea using a dryer for 5 minutes.
- the aqueous solution of 0.3% by weight HA with a weight average molecular weight of 600,000 to 1,200,000 as the control substance or the aqueous solution of 0.5% by weight cinnamate derivative-introduced HA prepared in the above Example (1-2) as the subject substance was administered to two corneas.
- the 1 ml injection syringes were used.
- Three rabbits (6 corneas) described in the above 1) were used as the administration subjects.
- the water evaporation amount was measured using a water evaporation amount measurement apparatus (AS-TW2, ASAHIBIOMED) before the administration of the test substance, after the drying treatment, after the administration of the test substance and until 40 minutes with 10 minute intervals after the administration of the test substance.
- AS-TW2, ASAHIBIOMED a water evaporation amount measurement apparatus
- the results of measuring the water evaporation amount were shown in FIG. 14 . From FIG. 14 , the water evaporation amount was slightly higher in the HA aqueous solution which was the control substance than in the saline which was the negative control whereas the saline became the value close to 0 after 40 minutes. On the other hand, the water evaporation amount after the administration of the subject substance kept the high value even when 40 minutes passed over, thereby the clear moisturizing performance of the subject substance was confirmed.
- the rabbit was euthanized, and after removing the eyeball, an entire corneal layer was removed by incising along the sclera.
- the removed cornea was preserved in saline, and the corneal epithelia was fixed by placing it on the paraffin block and the False-tooth stability material just before the measurement (hereinafter described as “the cornea to be measured”).
- the cornea to be measured was given cold air by the dryer from the distance of 30 cm for 5 minutes, and left standing at a room temperature for one hour.
- the amount perceived as an unperceived evaporation amount was directly measured as the water evaporation amount from the cornea to be measured using the water evaporation amount measurement apparatus (AS-TW2).
- the results of measuring the water evaporation amount were shown in FIG. 15 . From FIG. 15 , the water evaporation amount was slightly higher in the HA aqueous solution which was the control substance than in the saline which was the negative control whereas the saline exhibited the value close to 0 after 40 minutes. On the other hand, the water evaporation amount of the administered subject substance kept the high value even when 40 minutes had passed over, thereby it was confirmed that the subject substance has a more persistent water retention property on the cornea compared with saline and the HA aqueous solution.
- the group administered with (A), the group administered with (B) and the group administered with (C) are referred to as PBS group, HA group and crosslinked cinnamate derivative-introduced HA group, respectively.
- PBS group group administered with (A)
- HA group group administered with (B)
- C group administered with (C)
- administration classification administration group composition
- seven hamsters were used for each of the PBS group, the HA group and the crosslinked cinnamate derivative-introduced HA group.
- the water evaporation amount in the inside of the oral cavity in the xerostomia model hamster was measured using the water evaporation system (Asahibiomed), and the water evaporation amount ratio was calculated when the measurement value immediately after making the xerostomia model hamster was 1. The higher this water evaporation amount ratio is, the more moisturized condition is maintained (degree of oral cavity dryness is low).
- the measurement was performed immediately after making the xerostomia model hamster, immediately after the administration, 10 minutes and 20 minutes after the administration.
- FIG. 16 The results of measuring the water evaporation amount are shown in FIG. 16 .
- circled numbers 1, 2, 3 and 4 on the horizontal axis represent data immediately after making the xerostomia model hamster, immediately after the administration, 10 minutes and 20 minutes after the administration, respectively.
- P represents a significance level.
- All of administration groups exhibited the water evaporation amount ratio of 3.3 to 4.5.
- the water evaporation amount ratio 10 minutes after the administration was 0.9 on average in the PBS group and 2.3 on average in the HA group. On the other hand, it was 3.2 on average in the crosslinked cinnamate derivative-introduced HA group, thus the higher water evaporation amount ratio than the PBS group and the HA group was shown.
- the water evaporation amount ratio 20 minutes after the administration was 0.9 on average in the PBS group and 1.3 on average in the HA group.
- it was 3.2 on average in the crosslinked cinnamate derivative-introduced HA group thus the extremely higher water evaporation amount ratio compared with the PBS group and the HA group was indicated.
- the water evaporation amount ratio in the crosslinked cinnamate derivative-introduced HA group was very stable regardless of the elapse of time. This indicated that the crosslinked cinnamate derivative-introduced HA stayed at the administered site for a long period of time and exerted a highly persistent effect.
- GAG into which the hydrophobic group is introduced via the binding chain including the cinnamate derivative-introduced HA and the crosslinked cinnamate derivative-introduced HA has been shown to be suitable for the application to the mucosa and capable of effectively treating the disorder in the mucosal epithelial layer by being applied to the mucosa. It has been also shown that the effect of the treatment is highly persistent.
- the mixture was stirred over night and a 376 mg/5 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added.
- the mixture was stirred over night and a 381 mg/5 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer.
- the value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.”
- the results of the healed area in each individual are shown in FIG. 17 , and the results of a healing rate in each individual are shown in FIG. 18 .
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling ⁇ Area at each time point (after 1 to 3 days)
- FIGS. 17 and 18 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution at all time points of days 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution.
- the corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- the rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- the area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer.
- the value obtained by subtracting the area of the peeled site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.”
- C8-L(a: control), C8-R(b), C16-L(c: control), C16-R(d) represents left eye to which saline was administrated as a control for C8-R, right eye to which 0.1% octylamine-introduced HA solution was administrated, left eye to which saline was administrated as a control for C16-R, right eye to which 0.1% hexadecylamine-introduced HA solution was administrated, respectively.
- C8 represents results of the above study using octylamine
- C16 represents results of the above study using hexadecylamine.
- the healed area and the healing rate were calculated as follows.
- Healed area Area after peeling ⁇ Area at each time point (after 1 to 3 days)
- FIGS. 19 and 20 it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% octylamine-introduced HA and hexadecylamine-introduced HA solution at all time points of days 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% octylamine-introduced HA or hexadecylamine-introduced HA solution.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Dermatology (AREA)
- Biomedical Technology (AREA)
- Ophthalmology & Optometry (AREA)
- Nutrition Science (AREA)
- Physiology (AREA)
- Urology & Nephrology (AREA)
- Otolaryngology (AREA)
- Pulmonology (AREA)
- Gynecology & Obstetrics (AREA)
- Endocrinology (AREA)
- Reproductive Health (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
An agent for applying to mucosa capable of persistently exerting a therapeutic effect on disorders such as inflammation and lesions in the mucosa even by a lower frequency of administration because the agent can stay at a diseased site for a long period of time by exhibiting a high staying property in a mucosal epithelial layer is provided, said agent for application to mucosa containing glycosaminoglycan (GAG) into which a hydrophobic group is introduced via a binding chain, as an active ingredient.
Description
- The present invention relates to an agent for applying to mucosa containing a hydrophobic group binding type glycosaminoglycan as an active ingredient, and a method for production thereof.
- Conventionally, as substances having healing effects on mucosal disorders such as inflammation and lesions, hyaluronic acid which is a representative glycosaminoglycan (hereinafter described as “GAG”) has been known (e.g., Patent Document 1). However, in mucosa in the cornea, oral cavity and nasal cavity, and conjunctiva which contacts with the external world, and mucosa in the urinary bladder, the mucosal surfaces are washed with secretions and excretions such as tear fluid, salivary fluid and urine, and foreign substances are removed. Thus derivatives of GAG which keep medicinal effects inherent in GAG and exert a high staying property in these mucosal tissues have been demanded.
- Meanwhile, photoreactive hyaluronic acid whose water-solubility has been increased by binding a photocrosslinking group such as cinnamic acid to hyaluronic acid and further giving an alkali treatment thereto has been known (e.g., Patent Document 2). This photoreactive hyaluronic acid has been provided by binding a photocrosslinking group such as cinnamic acid to hyaluronic acid in order to provide medical materials such as anti-adhesion materials by giving the photocrosslinking, and does not aim at enhancing the staying property in the mucosal tissue.
- [Patent Document 1] Japanese Published Unexamined Patent Publication No. Hei-1-238530
- [Patent Document 2] Japanese Published Unexamined Patent Publication No. 2002-249501
- The present invention aims at providing an agent for applying to mucosa, which exerts an excellent staying property and pharmacological effects in mucosa.
- As a result of an extensive study for solving the above problem, the present inventors have found that “hydrophobic group binding type GAG” obtained by binding a hydrophobic group to GAG via a binding chain can be used as an extremely excellent active ingredient in an agent for applying to mucosa because this GAG keeps healing effects inherent in GAG on mucosal disorders such as inflammation and lesions and exhibits a high staying property when applied thereto, and have completed the present invention.
- The present invention relates to an agent for applying to mucosa which contains glycosaminoglycan (GAG) into which the hydrophobic group is introduced via the binding chain.
- The agent for applying to mucosa of the present invention can exert the persistent healing effect on the mucosal disorder such as inflammation and lesions even by low frequent administration because this can stay at a diseased site for a long period of time by exhibiting high staying property in the mucosa.
-
FIG. 1 is a view showing a spectrum of light transmittance; -
FIG. 2 is a view showing healed area percentages; -
FIG. 3 is a view showing healed areas; -
FIG. 4 is a view showing healing rate; -
FIG. 5 is a view showing healed areas; -
FIG. 6 is a view showing healing rate; -
FIG. 7 is a view showing healed areas; -
FIG. 8 is a view showing healing rate; -
FIG. 9 is a view showing healed areas; -
FIG. 10 is a view showing healing rate; -
FIG. 11 is a view showing staying property at peeled sites in rabbit corneal epithelia; -
FIG. 12 is a view showing staying property at peeled sites in rabbit corneal epithelia; -
FIG. 13 is a view showing photographs of an eyeball after irradiation with ultraviolet rays; -
FIG. 14 is a view showing amounts of water evaporation in a removed cornea; -
FIG. 15 is a view showing amounts of water evaporation in a removed cornea; and -
FIG. 16 is a view showing changes in water evaporation amount ratio in model hamsters for xerostomia. -
FIG. 17 is a view showing healed areas; -
FIG. 18 is a view showing healing rate; -
FIG. 19 is a view showing healed areas; -
FIG. 20 is a view showing healing rate. - The present invention will be described in more detail below by the best modes for carrying out the invention.
- Herein, an alkyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms. An alkenyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms, having at least one double bond. An alkynyl group refers to a straight or branched aliphatic hydrocarbon group having a described number of carbon atoms, having at least one triple bond.
- An aryl group refers to a monocyclic or polycyclic aromatic hydrocarbon group having 6 to 20 carbon atoms as ring-constituting atoms. A heteroaryl group refers to a monocyclic or polycyclic aromatic hydrocarbon group having 3 to 20 carbon atoms and one or more heteroatoms selected from nitrogen, sulfur and oxygen atoms as the ring-constituting atoms.
- An arylalkyl group refers to the alkyl group defined above substituted with the aryl group defined above. An arylalkenyl group refers to the alkenyl group defined above substituted with the aryl defined above. An arylalkynyl group is the alkynyl group defined above substituted with the aryl group defined above.
- An amino acid group refers to a group derived by losing a carboxyl group, an amino group or a hydroxyl group by a chemical bond from a natural or synthetic amino acid.
- Herein, the term “treatment” includes prevention, control of progression (prevention of deterioration), improvement (reduction) and cure of the mucosal disorder. The “mucosal disorder” means a condition where morphology, properties and functions to be inherent in the mucosa are disordered in some form. For example, the mucosal disorder can include the conditions such as lesions, defects, erosion, inflammation, ulcers and dryness.
- GAG into which the hydrophobic group is introduced via the binding chain, which is contained as the active ingredient in the agent for applying to mucosa of the present invention can be any GAG as long as the GAG binds the group having hydrophobicity derived from a hydrophobic compound having a water insoluble and oil soluble nature. This hydrophobic group is bound to GAG via the binding chain. As described later, it is not necessary that all constitutive units of GAG bind the hydrophobic groups.
- GAG in GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention is an acidic polysaccharide having a repeated long chain structure of disaccharide composed of amino sugar and uronic acid (or galactose). Examples of such GAG include hyaluronic acid, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan sulfate and keratan sulfate, and among them, hyaluronic acid is preferable. These GAG may be pharmaceutically acceptable salts thereof. Examples of such salts include sodium salts, potassium salts, magnesium salts and calcium salts, and among them, the sodium salt is preferable. Therefore, it is the most preferable that GAG in the agent for applying to mucosa of the present invention is sodium hyaluronate. An origin of GAG is not particularly limited, and GAG may be derived from an animal or a microorganism or chemically synthesized. For example, when using sodium hyaluronate, those derived from cock's comb can be exemplified. The molecular weight of GAG is not particularly limited, but its weight average molecular weight is preferably 200,000 to 3,000,000, more preferably 500,000 to 2,000,000 and most preferably 600,000 to 1,200,000. When hyaluronic acid or the pharmaceutically acceptable salt thereof is used, its weight average molecular weight is preferably 200,000 to 3,000,000, more preferably 500,000 to 2,000,000 and most preferably 600,000 to 1,200,000.
- The hydrophobic group in GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention is any group as long as the hydrophobic group is derived from the compound having the water insoluble and oil soluble nature. Examples of such a group can include alkyl groups having 2 to 18 carbon atoms, alkenyl groups having 2 to 18 carbon atoms, alkynyl groups having 2 to 18 carbon atoms, aryl groups, heteroaryl groups, arylalkyl groups, arylalkenyl groups, arylalkynyl groups and amino acid groups.
- The alkyl groups having 2 to 18 carbon atoms can include methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-butyl, n-pentyl, t-pentyl, isopentyl, neopentyl, n-heptyl, 5-methylhexyl, 4,4-dimethyl-pentyl, 1,1-dimethyl-pentyl and n-octyl. Among them, the alkyl groups such as n-butyl having 2 to 6 carbon atoms such as n-butyl can be preferably included.
- The alkenyl groups having 2 to 18 carbon atoms can include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-methyl-1-propenyl, 1-methyl-1-propenyl, 1-pentenyl, 3-methyl-2-butenyl, 1-heptene-1-yl and 2-heptene-1-yl. Among them, the alkenyl groups such as 1-butenyl having 2 to 6 carbon atoms such as 1-butenyl can be preferably included.
- The alkynyl groups having 2 to 18 carbon atoms can include ethynyl, 1-propinyl, 2-propinyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-heptynyl, 2-heptynyl and 3-heptynyl. Among them, the alkynyl groups such as 1-butynyl having 2 to 6 carbon atoms such as 1-butynyl can be preferably included.
- The aryl groups can include groups such as phenyl, naphthyl, anthryl and phenanthryl.
- The heteroaryl groups can include groups such as furyl, thionyl, thiazolyl, oxazolyl, pyrrolyl, pyridyl, pyrimidinyl and indolyl.
- The arylalkyl groups can include groups such as benzyl, phenethyl, naphthylmethyl and naphthylethyl.
- The arylalkenyl groups can include groups such as 2-phenyl-ethenyl and p-aminophenylethenyl.
- The arylalkynyl groups can include groups such as 2-phenyl-ethynyl and p-aminophenylethynyl.
- The amino acid groups can include groups derived from aliphatic amino acids such as glycine, alanine and β-alanine; branched aliphatic amino acids such as leucine, isoleucine and valine; aromatic amino acids such as phenylalanine and tyrosine; and heterocyclic amino acids such as tryptophan and histidine.
- These hydrophobic groups may be monosubstituted or polysubstituted with groups such as hydroxyl, carboxyl, cyano, amino (which may be monosubstituted or disubstituted with the above alkyl), nitro, oxo and alkylcarbonyloxy.
- Among the above hydrophobic groups, aryl groups, arylalkyl groups, arylalkenyl groups and arylalkynyl groups which are the hydrophobic groups containing the aryl group can be preferably included, and the arylalkenyl group and the aryl group substituted with the alkylcarbonyloxy group can be particularly preferably included. As such an arylalkenyl group, it is possible to specifically use phenylethenyl and p-aminophenylethenyl. As the aryl group, it is possible to preferably use the groups such as CH3—(CH2)l—COO-phenyl (wherein l represents 0 or an integer of 1 to 18).
- These hydrophobic groups may also have a function such as ultraviolet ray absorption ability due to having a double bond in the hydrophobic group as shown by a functional group such as phenyl-ethenyl exemplified above contained in the hydrophobic group. For example, when the agent for applying to mucosa of the present invention is used as eye drops described later, it is possible to make the eye drops having the function effectively absorbing the harmful ultraviolet rays by the use of the group having the ultraviolet ray absorption ability as the hydrophobic group. Furthermore, for example, when the agent for applying to mucosa of the present invention is used for the treatment of corneal epithelial layer disorders such as corneal xerosis (dry eye), keratoconjunctivitis, superficial punctate keratitis (SPK), corneal epithelial erosion, corneal epithelial loss and corneal tumor, it is possible to make the agent for applying to mucosa which has pharmacological effects on the above disorders in combination with the function effectively absorbing the harmful ultraviolet rays by the use of the group having the ultraviolet ray absorption ability as the hydrophobic group. As the group having the ultraviolet ray absorption ability, for example, arylalkenyl group having the conjugation double bond which is exemplified by 2-phenyl-ethenyl and p-aminophenylethenyl described above is preferable. When the agent for applying to mucosa of the present invention is used for the corneal disorder, it is preferable that “GAG into which the hydrophobic group is introduced” which is the active ingredient of the agent for applying to mucosa of the present invention is made into an aqueous solution of 0.1% by weight, which blocks 70 to 100% transmission of the ultraviolet rays at a wavelength of 200 to 300 nm when an ultraviolet ray transmittance is measured by the method described in the Example described later. Such a hydrophobic group having the ultraviolet ray absorption ability can preferably include the arylalkenyl groups such as 2-phenyl-ethenyl and p-aminophenylethenyl.
- In GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention, the above GAG is bound to the above hydrophobic group via the binding chain. GAG has the functional group which is a carboxyl, hydroxyl or sulfonate (—SO3H) group as the side chain. Thus, the hydrophobic group can be bound to GAG via the binding chain obtained by forming an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with these functional groups. Such a binding chain can specifically include —CONH—, —COO—, —O—, —SO3— and —SO2NH—. Among them, the carboxylic acid amide bond of —CONH— and the carboxylate ester bond of —COO— can be preferably used, and the carboxylic acid amide bond of —CONH— can be particularly preferably used.
- In GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention, the hydrophobic group is bound to GAG via the above binding chain, and a spacer chain may further exist between the binding chain and the hydrophobic group. As such a spacer chain, any chain group can be used as long as the spacer group does not completely lose the pharmacological effects which GAG has. Specifically, —(CH2)m— and —(CH2)—(OCH2)n— (wherein m and n are integers of 1 to 18, respectively) can be included.
- These spacer chains can further have the binding chains such as —CONH—, —COO—, —O—, —SO3— and —SO2NH— which are the same as above at the hydrophobic group side. Such a spacer chain having the binding chain at the hydrophobic group side can specifically include —COO—(CH2)m—, —COO—(CH2)—(OCH2)n—, —CONH—(CH2)m— and —CONH—(CH2)—(OCH2)n—.
- In GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention, it is not necessary that all of GAG constitutive units respectively have the hydrophobic groups. A ratio of the bound hydrophobic group in molar equivalent relative to a disaccharide repeat unit in molar equivalent of GAG (hereinafter, referred to as an “introduction ratio”) can be optionally determined depending on the type of the hydrophobic group, the degree of required hydrophobicity, the type of the mucosal disorder administered with the agent for applying to mucosa and the administration site, etc. For example, when using a phenylethenyl group which may be substituted as the hydrophobic group, preferably 5 to 30% and more preferably 10 to 20% of a hydrophobic group in molar equivalent is introduced relative to the disaccharide repeat unit in molar equivalent of GAG (in the case where the crosslinking bond described later is not formed).
- In GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention, the hydrophobic group may form a crosslinking bond between GAG molecules by the functional group contained in the group. As the hydrophobic group capable of forming the crosslinking bond, any group can be used as long as the hydrophobic group produces a photodimerization reaction or a photopolymerization reaction by irradiation of ultraviolet rays and is the same as defined above. The hydrophobic group capable of forming the crosslinking bond includes, for example, phenylethenyl, p-aminophenylethenyl, ethenyl, 2-carboxyethenyl and pentane-1,3-dienyl. It is desirable that these groups are bound to GAG via the binding chain which contains the carbonyl group. Among these hydrophobic groups, phenylethenyl or p-aminophenylethenyl which is bound to GAG via the binding chain which contains the carbonyl group can be particularly preferably used.
- In such GAG into which the hydrophobic group is introduced capable of forming the crosslinking bond, GAG molecules can be crosslinked with one another by being subjected to the photodimerization reaction or the photopolymerization reaction by standard methods. For example, according to the methods described in Japanese Published Unexamined Patent Publication No. 2002-249501, the photodimerization reaction or the photopolymerization reaction can be given.
- Representatives of GAG into which the hydrophobic group is introduced via the binding chain contained in the agent for applying to mucosa of the present invention can specifically include the agents for applying to mucosa containing GAG into which Ph-CH═CH—COO—(CH2)m—NHCO—; Ph-CH═CH—COO—CH2—(OCH2)n—NHCO—; Ph-CH═CH—CONH—(CH2)m—NHCO—; Ph-CH═CH—CONH—CH2—(OCH2)n—NHCO—; Ph-CH═CH—COO—(CH2)m—O—CO—; Ph-CH═CH—COO—CH2—(OCH2)n—O—CO—; Ph-CH═CH—CONH—(CH2)m—O—CO—; Ph-CH═CH—CONH—CH2—(OCH2)n—O—CO—; CH3—(CH2)l—COO-Ph-CONH—(CH2)m—NHCO— or CH3—(CH2)l—COO-Ph-CONH—CH2—(OCH2)n—NHCO— (wherein Ph represents phenyl group, m and n represent integers of 1 to 18, respectively, and l represents 0 or an integer of 1 to 18) is introduced, as the active ingredient.
- The following GAG can be included as the representative.
- GAG having the repeat unit of the structural unit represented by the
Chemical formula 1, as a basic skeleton: - wherein, R represents R1 or R2;
Ac represents an acetyl group;
R1 represents ONa or OH;
R2 represents (1) Ph-CH═CH—COO—(CH2)m—NH—;
(2) Ph-CH═CH—COO—CH2—(OCH2)n—NH—;
(3) Ph-CH═CH—CONH—(CH2)m—NH;
(4) Ph-CH═CH—CONH—CH2—(OCH2)n—NH—;
(5) Ph-CH═CH—COO—(CH2)m—O—;
(6) Ph-CH═CH—COO—CH2—(OCH2)n—O—;
(7) Ph-CH═CH—CONH—(CH2)m—O—;
(8) Ph-CH═CH—CONH—CH2—(OCH2)n—O—;
(9) CH3—(CH2)l—COO-Ph-CONH—(CH2)m—NH; or
(10) CH3—(CH2)l—COO-Ph-CONH—CH2—(OCH2)n—NH—;
wherein, Ph represents phenyl group, m and n represent integers of 1 to 18, respectively, and l represents 0 or an integer of 1 to 18, as a basic skeleton, wherein the ratio of the above structural unit wherein R represents R2 is 5 to 30% in molar equivalent relative to the disaccharide repeat unit in molar equivalent of the GAG.
8) Method for Producing GAG into which Hydrophobic Group is Introduced Via Binding Chain - To obtain GAG into which the hydrophobic group is introduced via the binding chain, GAG is reacted with a hydrophobic compound in which the above hydrophobic group has been bound to the functional group such as hydroxyl, carboxyl, amino or sulfonate group which can form an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the carboxyl, hydroxyl or sulfonate (—SO3H) group in GAG. Specifically, when the bond is the carboxylic acid amide bond, GAG having the carboxyl group is reacted with the hydrophobic compound having an amino group to bind the carboxyl group in GAG to the amino group in the hydrophobic compound. In the case of the carboxylate ester bond, GAG is reacted with the hydrophobic compound having hydroxyl or carboxyl group to bind the carboxyl group in GAG to the hydroxyl group in the hydrophobic compound or bind the hydroxyl group in GAG to the carboxyl group in the hydrophobic compound. In the case of the ether bond, GAG having the hydroxyl group is reacted with the hydrophobic compound having the hydroxyl group to react the hydroxyl group in GAG with the hydroxyl group in the hydrophobic compound. In the case of the sulfonate ester bond, GAG is reacted with the hydrophobic compound having the hydroxyl group or sulfonate group to bind the hydroxyl group in GAG to the sulfonate group in the hydrophobic compound or bind the sulfonate group in GAG to the hydroxyl group in the hydrophobic compound. These reactions can be performed by common standard methods, and reaction conditions can be optionally selected by those skilled in the art.
- When the spacer chain is present between the binding chain and the hydrophobic group, the order in introducing the spacer chain and the hydrophobic group to GAG is not particularly limited. For example, either the method in which a spacer compound having the functional group such as the hydroxyl, carboxyl, amino or sulfonate group which can form the ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the functional group in GAG at one end of the above spacer chain is reacted with GAG, and subsequently, the other end of the spacer compound is reacted with the hydrophobic compound which is bound to the functional group such as the hydroxyl, carboxyl, amino or sulfonate group, or the method in which the spacer compound having the functional group such as the hydroxyl, carboxyl, amino or sulfonate group which can form an ether bond, carboxylate ester bond, sulfate ester bond, carboxylic acid amide bond or sulfonate amide bond together with the functional group in the hydrophobic compound at one end is reacted with the hydrophobic compound in which the hydrophobic group has been bound to the functional group such as the hydroxyl, carboxyl, amino or sulfonate group, and subsequently the other end of the spacer compound is reacted with GAG may be used. In particular, the method in which the spacer compound is reacted with the hydrophobic compound followed by being reacted with GAG can be preferably used.
- The above-described method can be appropriately carried out by publicly known methods, and preferably performed in the presence of a condensing agent. Such a condensing agent can preferably include water soluble carbodiimide such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI.HCl), condensing agents such as dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinate imide (HOSu). For example, when hyaluronic acid is used as GAG and the cinnamate derivative such as Ph-CH═CH—COO—(CH2)m—NH2 or Ph-CH═CH—COO—CH2—(OCH2)n—NH2 (wherein m and n are integers of 1 to 18, respectively) is used as the hydrophobic compound which is bound to the spacer compound, the condensation method using water soluble carbodiimide such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) and N-hydroxysuccinate imide can be preferably used. The reaction can be accomplished using a mixed solvent of water and a water soluble organic solvent such as dioxane, dimethylformamide or ethanol. The hyaluronate derivative which is highly soluble in an aqueous vehicle can be obtained by treating with a base such as sodium hydrogen carbonate after the completion of the reaction.
- When the thus produced GAG into which the hydrophobic group is introduced via the binding chain is subjected to the photodimerization reaction or the photopolymerization reaction to cross link the GAG molecules with one another, for example, the method described in Japanese Published Unexamined Patent Publication No. 2002-249501 can be used. Specifically, in the case of the compound in which the phenylethenyl group as the hydrophobic group is bound to GAG via —COO—(CH2)m—NHCO—, the crosslink can be formed by irradiating light to the solution containing them using an ultraviolet lamp.
- The agent for applying to mucosa of the present invention contains one or more GAG into which the hydrophobic group is introduced via the binding chain as the active ingredient, and may also further include other medically, pharmaceutically or biologically acceptable substances other than the GAG into which the hydrophobic group is introduced via the binding chain. Such substances include but are not limited to, salts such as sodium chloride, potassium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate and monopotassium hydrogen phosphate, and preservatives such as paraoxybenzoate esters, benzalkonium chloride, chlorobutanol and chlorhexidine gluconate, and other pharmacologically active ingredients.
- The agent for applying to mucosa of the present invention can be made into any publicly known formulation forms (e.g., solid preparations such as granules and powder, liquid preparations such as aqueous solutions, suspension and emulsion, and gel preparations) as the pharmaceutical for applying to the mucosa. In the agent for applying to mucosa of the present invention, the form thereof upon formulating and distributing and the form thereof upon applying to the mucosa may be the same or different. For example, the agent for applying to mucosa of the present invention may be formulated in the form of solution and may be applied directly to the mucosa as it is. Also, the agent for applying to mucosa of the present invention may be formulated and distributed in the solid form, and may be made into solution or gel when being applied to the mucosa. Thus, the agent for applying to mucosa of the present invention can be made into the formulation form for being prepared when used.
- When being made into the liquid agent by dissolving in water, the amount of GAG into which the hydrophobic group is introduced via the binding chain is preferably 0.02 to 5% by weight, more preferably 0.1 to 3% by weight, extremely preferably 0.1 to 1% by weight and most preferably 0.1 to 0.6% by weight.
- The agent for applying to mucosa of the present invention aims at applying to the mucosa. Animals to which the agent for applying to mucosa of the present invention is applied are not particularly limited as long as they have the mucosa, and mammalian animals are preferable. The mammalian animals include, but are not limited to, humans, horses, cattle, dogs, cats, rabbits, hamsters, guinea pigs and mice. The agent for applying to mucosa of the present invention may be of course made into the pharmaceuticals for humans, and can also be made into the pharmaceuticals for the animals. Among them, it is preferable to be made into the pharmaceuticals for humans.
- The mucosa to which the agent for applying to mucosa of the present invention can be applied is not particularly limited as long as the mucosa is the mucosa present in the animal. Such mucosa include mucosal tissues present in organs and tissues exemplified by the gastrointestinal system such as the stomach and intestines, the cardiovascular system, the respiratory system, the excretion system such as the urinary bladder, rectum and anus, the genital system such as the vagina, and organs such as eyes, nose and oral cavity which contact with the external world. Among them, the agent for applying to mucosa of the present invention can be preferably applied to the cornea, conjunctiva, oral cavity mucosa and urinary bladder mucosa.
- The agent for applying to mucosa of the present invention can be widely applied to such mucosa. The purpose of the application is not particularly limited, and for example, the purposes such as protection of the mucosal tissue (e.g., prevention of snow blindness by ultraviolet rays, pterygium and cataract), prevention of mucosal dryness and the treatment of mucosal disorder can be exemplified. Thus, the agent for applying to mucosa of the present invention can be applied to not only the mucosa in the abnormal state (e.g., mucosa where the disorder has occurred) but also the mucosa in the normal state. However, since the agent for applying to mucosa of the present invention exerts excellent pharmacological effects in the mucosa where the disorder has occurred, it is possible to preferably use for the treatment of the mucosal disorders, e.g., disorders in the cornea, conjunctiva, oral cavity mucosa and urinary bladder mucosa.
- Since the agent for applying to mucosa of the present invention exerts excellent pharmacological effects particularly on disorders in the mucosal epithelia among the mucosal disorders, it is possible to be preferably used for the treatment of the disorders in the mucosal epithelia.
- Examples of such disorders in the mucosal epithelia include corneal epithelial layer disorders such as corneal xerosis (dry eye); keratoconjunctivitis, superficial punctate keratitis (SPK), corneal epithelial erosion, corneal epithelial loss and corneal tumor; oral cavity mucosal disorders such as xerostomia (dry mouth), aphthous ulcer, stomatitis and glossitis; dryness and pruritus of nasal mucosa; urinary bladder mucosal disorders such as interstitial cystitis; ulcerative proctitis, and dryness of the rectum or vagina. Also dryness and lesions of organ mucosa upon surgical operation can be exemplified. Among them, it is possible to be preferably used for the treatment of the corneal epithelial layer disorders, the oral cavity mucosal epithelial disorders and the urinary bladder mucosa epithelial layer disorders.
- The agent for applying to mucosa of the present invention can be applied to the mucosal tissues exemplified above, and its application method and application formulation can be appropriately determined by those skilled in the art depending on the position, morphology, property and function of the mucosa to be applied, and the purpose of the application. However, it is preferable that the agent for applying to mucosa of the present invention is applied to the mucosa in the liquid form such as solution in use. In that case, upon producing (formulating) or applying the agent for applying to mucosa of the present invention, the liquid can be obtained by dissolving GAG into which the hydrophobic group is introduced via the binding chain in the solvent. The solvent is not particularly limited as long as the solvent can dissolve the GAG into which the hydrophobic group is introduced via the binding chain and is the pharmaceutically acceptable solvent. For example, a buffer such as a phosphate buffer or saline can be used, but the solvent is not limited thereto. In this case, the concentration of the GAG into which the hydrophobic group is introduced via the binding chain in the liquid agent is not particularly limited, and can be appropriately determined depending on the type of the mucosa to be applied and the degree of the mucosal disorder. When the agent for applying to mucosa of the present invention is the eye drops, when the agent for applying to mucosa of the present invention is applied to the oral cavity mucosa or the urinary bladder mucosa, for example, the concentration is preferably 0.02 to 5% by weight, more preferably 0.1 to 3% by weight, still more preferably 0.1 to 1% by weight, still more preferably 0.1 to 0.6% by weight, extremely preferably 0.1 to 0.5% by weight and most preferably 0.1 to 0.3% by weight.
- When the agent for applying to mucosa of the present invention is applied to the mucosa in the stomach as the liquid as above, an oral administration or the administration using a catheter can be selected. When applied to the mucosa in the eye, the nose or the oral cavity, for example, the administration method such as instillation of drop, nasal instillation or oral inclusion can be selected. For example, when the agent for applying to mucosa of the present invention is applied to the mucosa mucosa in the urinary bladder, rectum or vagina, or the mucosa of organs where the dryness is concerned upon surgical operation, the method of administering by injecting, spraying or applying the agent for applying to mucosa of the present invention to a lumen or a surface of these organs or tissues can be selected, but the methods are not limited thereto.
- The amount, the number of times and the frequency of the application (administration of) of the agent for applying to mucosa of the present invention is not particularly limited, and should be determined depending on the mucosa subjected to the application, the purpose of the application, the type, age, body weight, gender, and degree of mucosal disorder in the animal to be applied.
- Specifically, when the agent for applying to mucosa of the present invention is used for the purpose of treating the human corneal epithelial layer disorder, the agent for applying to'mucosa of the present invention at the above-described concentration as the liquid formulation for the instillation of drops (eye drops) containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by instilling 1 to 3 drops per
administration 1 to 5 times per day, and may be administered by instilling 1 to 3 drops peradministration 1 to 3 times per day. - When the agent for applying to mucosa of the present invention is used for the purpose of treating the human oral cavity mucosal disorder, the agent for applying to mucosa of the present invention at the above-described concentration as the liquid containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by putting the agent for applying to mucosa of the present invention in the
oral cavity 1 to 5 times per day and rinsing for approximately several tens of seconds (preferably approximately 20 to 30 seconds) followed by spitting it out. - When the agent for applying to mucosa of the present invention is applied to the urinary bladder mucosal disorder, this is preferably used for the treatment of the urinary bladder mucosal disorders exemplified by non-bacterial refractory cystitis exemplified by interstitial cystitis, eosinophilic cystitis and hemorrhagic cystitis which do not respond to anti-bacterial agents although symptoms similar to those of acute bacterial cystitis are exhibited. In this case, the agent for applying to mucosa of the present invention at the above-described concentration as the liquid containing GAG into which the hydrophobic group is introduced via the binding chain can be administered by administering the agent for applying to mucosa of the present invention directly to the urinary bladder at the amount of 50 mL per
administration 1 to 7 times per week or administering with a catheter in the urinary bladder. - The agent for applying to mucosa of the present invention can stay at the diseased site for a longer period of time because the active ingredient contained in the agent exhibits the high staying property in the mucosa, compared with the conventional drugs containing hyaluronic acid as the active ingredient in which no hydrophobic group has been bound. Therefore, the agent for applying to mucosa of the present invention can also exert the treating effect persistently even at the low administration frequency on the disorders such as inflammation and lesions in the mucosa. However, the agent for applying to mucosa of the present invention is not limited by its administration frequency.
- The present invention will be described below by Examples.
- A 172 mg/5 mL aqueous solution of N-hydroxysuccinimide (HOSu: Watanabe Chemical Industries, Ltd.), a 143 mg/5 mL aqueous solution of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI.HCl) (Watanabe Chemical Industries, Ltd.), and a 181 mg/5 mL aqueous solution of 3-aminopropyl cinnamate hydrochloride (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.06 g, 2.7° mmol/disaccharide unit, weight-average molecular weight 900,000; derived from cock's comb, Seikagaku Corporation) in water (115 mL)/dioxane (144 mL). The mixture was stirred for 3 hours and a 750 mg/10 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 2
h 30 min., the reaction was quenched with acetic acid (214 mg) and sodium chloride (1.0 g). Ethanol (300 mL) was added and the resulting precipitation was filtered off and washed twice successively by 80% ethanol, 95% ethanol. The solid was dried in vacuo at 40° C. over night to afford white solid (1.06 g) (hereinafter in Examples, “cinnamate derivative-introduced sodium hyaluronate” is abbreviated as “cinnamate derivative-introduced HA”). The introduction ratio of the cinnamate derivative was 16%. The introduction ratio of the cinnamate derivative was calculated based on the amount of cinnamate by an absorbance measurement method (wavelength: 269 nm) and the amount of hyaluronate by a carbazole sulfate method. - Saline was added to 86 mg of cinnamate derivative-introduced HA obtained in the above (1-1) to give a total amount of 15.45 ml, then the solution was shaken over night with a shaker until uniformly dissolved. The 0.5% by weight solution of cinnamate derivative-introduced HA (in drying
loss 10%) was obtained. Likewise, the 0.3% by weight and 0.1% by weight solutions of cinnamate derivative-introduced HA were obtained. - A 75 mg/5 mL aqueous solution of HOSu, a 62 mg/5 mL WFI solution of EDCI.HCl, and a 92 mg/5 mL WFI solution of 6-aminohexyl cinnamate hydrochloride (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.0 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 1,500,000; derived from cock's comb, Seikagaku Corporation) in water for injection (hereinafter referred to as WFI) (150 mL)/dioxane (75 mL). The mixture was stirred for 4 hours and sodium chloride (1.0 g) was added. Ethanol (500 mL) was added and the resulting precipitation was filtered off and washed twice successively by 80% ethanol, ethanol. The solid was dried in vacuo at 40° C. to afford white solid (1.1 g). The introduction ratio of the cinnamate derivative was 2.7%.
- The above cinnamate derivative-introduced HA (12.5 g) was dissolved in phosphate buffered saline (concentration of phosphate: 1.5 mM, hereinafter abbreviated as “PBS”) to prepare 2.5% solution of cinnamate derivative-introduced HA (500 mL). The 2.5% solution of cinnamate derivative-introduced HA was irradiated by 800 W high pressure mercury lamp and performed by a heat treatment in an autoclave at 121° C. for 7.5 min. to yield crosslinked cinnamate derivative-introduced HA.
- Further, 1 g of the above crosslinked cinnamate derivative-introduced HA was dissolved in 11.5 ml of WFI to prepare the 0.2% by weight of crosslinked cinnamate derivative-introduced HA.
- A 172 mg/5 mL aqueous solution of HOSu, a 143 mg/5 mL aqueous solution of EDCI.HCl, and a 181 mg/5 mL aqueous solution of 3-aminopropyl cinnamate hydrochloride (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.0 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 900,000; derived from cock's comb, Seikagaku Corporation) in water (150 mL)/dioxane (75 mL). The mixture was stirred for 3 hours and a 750 mg/10 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 2
h 30 min., the reaction was quenched with acetic acid (214 mg) and sodium chloride (1.0 g). Ethanol (300 mL) was added and the resulting precipitation was filtered off and washed twice successively by 80% ethanol, 95% ethanol. The solid was dried in vacuo at 40° C. to afford white solid (1.0 g) as cinnamate derivative-introduced HA. The introduction ratio of the cinnamate derivative was 10.1%. - A 3.0 mmol/mL aqueous solution of HOSu, a 1.5 mmol/mL aqueous solution of EDCI.HCl and a 1.5 mmol/mL aqueous solution of 4-aminofluorescein (Tokyo Chemical Industry Co., Ltd.) were added to a solution of cinnamate derivative-introduced HA obtained in the above (3-1) (1.00 g, 2.5 mmol/disaccharide unit) in water (150 mL)/dioxane (75 mL). The mixture was stirred one day and a 500 mg/10 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 4
h 30 min., the reaction was quenched with acetic acid (2 mL) and sodium chloride (6.0 g). Ethanol (500 mL) was added and the resulting precipitation was filtered off and washed four times by 80% ethanol, twice by ethanol. The solid was dried in vacuo over night to afford fluorochrome-labeled solid (782 mg) The introduction ratio of the fluorescence was 0.60%. - A 3.0 mmol/mL aqueous solution of HOSu, a 1.5 mmol/mL aqueous solution of EDCI.HCl and a 1.5 mmol/mL aqueous solution of 4-aminofluorescein (Tokyo Chemical Industry Co., Ltd.) were added to a solution of sodium hyaluronate (1.00 g, 2.5 mmol/disaccharide unit, weight-average molecular weight 900,000; derived from cock's comb, Seikagaku Corporation) in water (150 mL)/dioxane (75 mL). The mixture was stirred one day and a 500 mg/10 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 4
h 30 min., the reaction was quenched with acetic acid (2 mL) and sodium chloride (6.0 g). Ethanol (500 mL) was added and the resulting precipitation was filtered off and washed four times by 80% ethanol, twice by ethanol. The solid was dried in vacuo over night to afford fluorochrome-labeled solid (830 mg) The introduction ratio of the fluorescence was 0.32%. - The 0.1% by weight aqueous solution of cinnamate derivative-introduced HA obtained in the above (1-1) was prepared, and the ultraviolet ray transmittance was measured by an spectrometer (UV-1600, Shimadzu Corporation).
- A spectrum which indicates the transmittance is shown in
FIG. 1 , and the transmittance (%) at various wavelengths is shown in Table 1. As a result, 100% of the transmittance was shown at the wavelengths of 340 nm or more, but the transmittance at the wavelengths of approximately 320 nm or less was 20% or less which was extremely low, and it was demonstrated that this solution effectively blocks the transmission of the ultraviolet ray. - In
FIG. 1 , scales are shown with 65 nm intervals on a horizontal axis and with 20% intervals on a vertical axis. -
TABLE 1 λ T (%) 234 19.9 340 106.2 380 101.8 450 98.5 In the table, λ and T represent the wavelength and the transmittance (%), respectively. - The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after the corneal epithelia were peeled, 150 μl of saline as the control substance was administered in the left eye, and 150 μl of 0.5% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. On one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hours interval, the same administration as above was performed. In the administration, 1 ml injection syringes were used. Six model rabbits for the corneal epithelial layer disorder described in the above 1) were used as administration subjects.
- 3) Photographing of Corneal Epithelial Defective Region
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the final administration at 3 days after the peeling. When photographed, a focal length was made constant to make a magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using an image analyzer. A value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after peeling the corneal epithelia was performed was rendered as “healed area.” - The results of a healed area percentage in each individual are shown in
FIG. 2 , and the results of the healed area percentage and a healed area percentage ratio in each individual are shown in Table 2. The healed area percentage and the healed area percentage ratio were calculated as follows. -
Healed area percentage (%)=(Healed area/Peeled area)×100 -
Healed area percentage ratio=(Healed area percentage in right eye/Healed area percentage in left eye)×100 -
TABLE 2 Healed area percentage (%) right eye (administration of cinnamate left eye Healed area Specimen derivative- (administration percentage number introduced HA) of saline) ratio 1 76.90 66.59 115.47 2 69.44 85.90 80.85 3 75.23 61.71 121.91 4 75.54 56.79 133.01 5 68.41 64.53 106.01 6 83.76 66.92 125.16 Mean 74.88 67.07 113.73 Standard 5.57 9.95 — deviation - For each individual of
individual numbers 1 to 6, the left column shows the healed area percentage in the right eye (administration of cinnamate derivative-introduced HA), and the right column shows the healed area percentage in the left eye (administration of saline). InFIG. 2 and Table 2, the apparent effect to facilitate the healing of the corneal epithelial layer disorder was observed in 5 of the administered 6 individuals. - The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after the corneal epithelia were peeled, 150 μl of saline as the control substance was administered in the left eye, and 150 μl of 0.5% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. On one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hour intervals, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Fourteen model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.” - The results of the healed area in each individual are shown in
FIG. 3 , and the results of a healing rate in each individual are shown inFIG. 4 . The healed area and the healing rate were calculated as follows. -
Healed area=Area after peeling*−Area at each time point (after 1 to 3 days) - *Herein after, “area after peeling” means ‘just before the administration of the subject substance one hour after the cornea was peeled’ in calculation of healed area.
-
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 3 and 4 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.5% by weight cinnamate derivative-introduced HA solution, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.5% by weight cinnamate derivative-introduced HA solution. - The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after peeling the corneal epithelia, 150 μl of saline as the control substance was administered in the left eye, and 150 μl of 0.3% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. At one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hour intervals, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Fourteen model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.” - The results of the healed area in each individual are shown in
FIG. 5 , and the results of a healing rate in each individual are shown inFIG. 6 . The healed area and the healing rate were calculated as follows. -
Healed area=Area after peeling−Area at each time point (after 1 to 3 days) -
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 5 and 6 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.3% by weight cinnamate derivative-introduced HA solution at all time points ofdays 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.3% by weight cinnamate derivative-introduced HA solution. - The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after peeling the corneal epithelia, 150 μl of an aqueous solution of 0.1% by weight HA with a weight average molecular weight 600,000 to 1,200,000 as the control substance was administered in the left eye, and 150 μl of 0.1% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. At one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hour intervals, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Eight model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.” - The results of the healed area in each individual are shown in
FIG. 7 , and the results of the healed area and the healing rate in each individual are shown inFIG. 8 . The healed area and the healing rate were calculated as follows. -
Healed area=Area after peeling−Area at each time point (after 1 to 3 days) -
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 7 and 8 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution, compared with the healed area of the corneal epithelia in the control eyes administered with the 0.1% by weight HA aqueous solution. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution. - The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour after the corneal epithelia were peeled, 150 μl of the aqueous solution of 0.1% by weight HA with a weight average molecular weight 600,000 to 1,200,000 as the control substance was administered in the left eye, and 150 μl of 0.1% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. Furthermore, at one to 3 days after the peeling, once a day, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Eight model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 6 hours after the administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at, 3′ days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as the “healed area.” - The results of the healed area in each individual are shown in
FIG. 9 , and the results of the healing rate in each individual are shown inFIG. 10 . The healed area and the healing rate were calculated as follows. -
Healed area=Area after peeling−Area at each time point (after 1 to 3 days) -
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 9 and 10 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution at all time points of thedays 1 to 3, compared with the healed area of the corneal epithelia in the control eyes administered with the 0.1% by weight HA aqueous solution. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution. - The effect of the fluorescence labeled cinnamate derivative-introduced HA prepared in Example 3 and the fluorescence labeled HA prepared in Comparative Example 1 on the residual property of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour after the corneal epithelia were peeled, 150 μl of the aqueous solution of 0.3% by weight fluorescence labeled HA prepared in the above Comparative Example 1 as the control substance was administered in the left eye, and 150 μl of the aqueous solution of 0.3% by weight fluorescence labeled cinnamate derivative-introduced HA prepared in the above Example 3 as the subject substance was administered in the right eye. In the administration, the 1 ml injection syringes were used. Eight model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- Two rabbits were given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine per rabbit, and eyeballs were removed 30 minutes, one hour, one and a half hours and 2 hours and 30 minutes after the administration of the subject substance and the control substance. A pore was opened between the cornea and sclera in the removed eyeball using a surgical knife, and only the cornea was taken out using the microscissors. The removed cornea was placed on a biological sample slicing plate (supplied from Nisshin EM Corporation, Cat No. 428), and the portion to be observed was cut out using a single-edged razor blade stainless steel (GEM® STAINLESS STEEL UNCOATED, Nisshin EM Corporation, Cat No. 429). The cut out portion was immersed in O.C.T. compound (Tissue-Tek® 4583, Lot. 1178), then embedded in a cryostat tray (supplied by Murazumi Co., Ltd., Cat. No. 31) filled with the O.C.T. compound so that the portion to be observed was at a bottom, and rapidly frozen using liquid nitrogen in foam polystyrene to make an unfixed frozen block.
- Subsequently, the frozen block was removed from the cryostat tray, and attached on a sample table using the O.C.T. compound. The sample table and a disposable microtome blade (supplied by Leica Microsystems Japan, Model 818, Lot. No. 913212) were set in a high performance frozen microtome for research, and the block was sliced under the condition of a frozen chamber temperature (CT) at −20° C. and sample side temperature (OT) at −16° C. to make sections with a thickness of 5 μm using silane coating slide glasses (supplied by Muto Pure Chemicals Co., Ltd., Star Frost Slide Glass, Cat. No. 5116).
- The frozen section was set in an incident-light fluorescence microscope (Olympus Corporation, BH2-RFC), FA images and autofluorescent images were observed at IB cube (BH2-DMIB, excitation wavelength: 495 nm, absorption wavelength: 460 nm) and U cube (BH2-DMU, broad band U excitation, absorption wavelength: 435 nm), respectively. The FA image and the autofluorescent image were photographed using a cooled high sensitivity CCD camera (Keyence Corporation, VB-6010) under the condition of exposure time for one second and ISO sensitivity of 200.
- Photographs of the sampled cornea were shown in
FIGS. 11 and 12 . FromFIGS. 11 and 12 , it was identified by color development of the fluorescence label that the aqueous solution of 0.3% by weight fluorescence labeled HA which was the control substance stayed until 30 minutes after the administration but did not stay after one hour. Meanwhile, although the fluorescent color development of the aqueous solution of 0.3% by weight fluorescence labeled cinnamate derivative-introduced HA which was the subject substance was weakened with the elapse of time, the color development was observed at all time points from 30 minutes to 2 hours and 30 minutes after the administration, thereby the high staying performance was confirmed. - The protective effect on the cinnamate derivative-introduced HA prepared in Example 1 on the rabbit with corneal superficial punctuate keratopathy.
- Introduced anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine and maintained anesthesia by inhalation of isoflurane were given to the rabbit. Subsequently, the eyelid was always opened using an eyelid retractor for kids.
- In the condition where the eye was opened, 150 μl of an aqueous solution of 0.3% by weight HA with a weight average molecular weight 600,000 to 1,200,000 as the control substance was administered in the right eye, and 150 μl of 0.3% by weight cinnamate derivative-introduced HA prepared in the above Example (1-2) as the subject substance was administered in the left eye. In the administration, the 1 ml injection syringes were used. One rabbit described in the above 1) was used as the administration subject.
- Ultraviolet rays were irradiated to both eyes from a distance of approximately 10 cm apart from the rabbit eyeball using a 15 kW germicidal lamp. The irradiation was performed for 3 hours.
- The eyeball was stained with 0.2% sodium fluorescein under the continuous anesthesia of the rabbit, and photographed under violet light. When photographed, the focal length was made constant to make the magnification of photographs constant.
- Photographs after the irradiation of the ultraviolet rays were shown in
FIG. 13 . FromFIG. 13 , in the eyeball irradiated with the ultraviolet rays after the administration of the control substance, the disordered site stained with 0.2% sodium fluorescein was apparent. Meanwhile, in the eyeball irradiated with the ultraviolet rays after the administration of the subject substance, the disordered site stained with 0.2% sodium fluorescein was clearly smaller than that of the control substance, and the corneal disorder caused by the ultraviolet ray was prevented. - Using the removed cornea of the rabbit, the moisturizing performance of the cinnamate derivative-introduced HA prepared in Example 1 was validated.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, and the eyeball was removed. A pore was opened between the cornea and the sclera in the removed-eyeball using a surgical knife, and only the cornea was taken out using the microscissors.
- The drying treatment was performed by placing the removed cornea on a paraffin block with a False-tooth stability material and providing cold air from the distance of approximately 1 m from the cornea using a dryer for 5 minutes.
- After the completion of the drying treatment, 100 μl of saline as the negative control substance, the aqueous solution of 0.3% by weight HA with a weight average molecular weight of 600,000 to 1,200,000 as the control substance or the aqueous solution of 0.5% by weight cinnamate derivative-introduced HA prepared in the above Example (1-2) as the subject substance was administered to two corneas. In the administration, the 1 ml injection syringes were used. Three rabbits (6 corneas) described in the above 1) were used as the administration subjects.
- The water evaporation amount was measured using a water evaporation amount measurement apparatus (AS-TW2, ASAHIBIOMED) before the administration of the test substance, after the drying treatment, after the administration of the test substance and until 40 minutes with 10 minute intervals after the administration of the test substance.
- The results of measuring the water evaporation amount were shown in
FIG. 14 . FromFIG. 14 , the water evaporation amount was slightly higher in the HA aqueous solution which was the control substance than in the saline which was the negative control whereas the saline became the value close to 0 after 40 minutes. On the other hand, the water evaporation amount after the administration of the subject substance kept the high value even when 40 minutes passed over, thereby the clear moisturizing performance of the subject substance was confirmed. - The rabbit was euthanized, and after removing the eyeball, an entire corneal layer was removed by incising along the sclera. The removed cornea was preserved in saline, and the corneal epithelia was fixed by placing it on the paraffin block and the False-tooth stability material just before the measurement (hereinafter described as “the cornea to be measured”).
- The cornea to be measured was given cold air by the dryer from the distance of 30 cm for 5 minutes, and left standing at a room temperature for one hour.
- After the water is evaporated, two drops (approximately 100 μl) of saline as the negative control substance, the aqueous solution of 0.3% by weight HA with a weight average molecular weight of 600,000 to 1,200,000 as the control substance or the aqueous solution of 0.5% by weight cinnamate derivative-introduced HA prepared in the above Example (1-2) as the subject substance was administered by the 1 ml syringe.
- The amount perceived as an unperceived evaporation amount (released water amount per m2 per hour) was directly measured as the water evaporation amount from the cornea to be measured using the water evaporation amount measurement apparatus (AS-TW2).
- The results of measuring the water evaporation amount were shown in
FIG. 15 . FromFIG. 15 , the water evaporation amount was slightly higher in the HA aqueous solution which was the control substance than in the saline which was the negative control whereas the saline exhibited the value close to 0 after 40 minutes. On the other hand, the water evaporation amount of the administered subject substance kept the high value even when 40 minutes had passed over, thereby it was confirmed that the subject substance has a more persistent water retention property on the cornea compared with saline and the HA aqueous solution. - Using a model hamster for xerostomia, the healing effect of the crosslinked cinnamate derivative-introduced HA prepared in Example 2 on the xerostomia was validated.
- An inside of the oral cavity of a male. Syrian hamster was exposed by inserting a test tube with a diameter of approximately 10 mm in the oral cavity close to a buccal side and reversing it under the anesthesia with Nembutal. By giving hot air for approximately 20 seconds, and subsequently giving cold air for 2 minutes and 40 seconds to the exposed inside of the oral cavity using a dryer, the model hamster for the xerostomia was obtained. The inside of the oral cavity was continuously exposed until the measurement was completed.
- Immediately after making the xerostomia model, 1.00 μl of (A) PBS, (B) 0.2% by weight HA solution (supplied by Seikagaku Corporation, weight average molecular weight: 1,500,000) or (C) 0.2% by weight crosslinked cinnamate derivative-introduced HA solution was administered by applying to the inside of the oral cavity using a microsyringe.
- Hereinafter, the group administered with (A), the group administered with (B) and the group administered with (C) are referred to as PBS group, HA group and crosslinked cinnamate derivative-introduced HA group, respectively. For the administration classification (administration group composition), seven hamsters were used for each of the PBS group, the HA group and the crosslinked cinnamate derivative-introduced HA group.
- The water evaporation amount in the inside of the oral cavity in the xerostomia model hamster was measured using the water evaporation system (Asahibiomed), and the water evaporation amount ratio was calculated when the measurement value immediately after making the xerostomia model hamster was 1. The higher this water evaporation amount ratio is, the more moisturized condition is maintained (degree of oral cavity dryness is low). The measurement was performed immediately after making the xerostomia model hamster, immediately after the administration, 10 minutes and 20 minutes after the administration.
- The results of measuring the water evaporation amount are shown in
FIG. 16 . In the figure, circlednumbers evaporation amount ratio 10 minutes after the administration was 0.9 on average in the PBS group and 2.3 on average in the HA group. On the other hand, it was 3.2 on average in the crosslinked cinnamate derivative-introduced HA group, thus the higher water evaporation amount ratio than the PBS group and the HA group was shown. Further, the waterevaporation amount ratio 20 minutes after the administration was 0.9 on average in the PBS group and 1.3 on average in the HA group. On the other hand, it was 3.2 on average in the crosslinked cinnamate derivative-introduced HA group, thus the extremely higher water evaporation amount ratio compared with the PBS group and the HA group was indicated. Furthermore, as is apparent fromFIG. 16 , the water evaporation amount ratio in the crosslinked cinnamate derivative-introduced HA group was very stable regardless of the elapse of time. This indicated that the crosslinked cinnamate derivative-introduced HA stayed at the administered site for a long period of time and exerted a highly persistent effect. - From the above results, GAG into which the hydrophobic group is introduced via the binding chain including the cinnamate derivative-introduced HA and the crosslinked cinnamate derivative-introduced HA has been shown to be suitable for the application to the mucosa and capable of effectively treating the disorder in the mucosal epithelial layer by being applied to the mucosa. It has been also shown that the effect of the treatment is highly persistent.
- Since no adverse effect due to the administration of the cinnamate derivative-introduced HA and the crosslinked cinnamate derivative-introduced HA was observed in any of the above animal studies, the safety of the agent for applying to mucosa of the present invention can be sufficiently estimated.
- A 25.8 mg/2 mL solution (ethanol:0.1MHCl=1:1) of octylamine, 2 mL of 0.1 M solution (ethanol:water=1:1) of DMT-MM (Wako Pure Chemical Industries, Ltd.) were added to a solution of sodium hyaluronate (502 mg, 1.25 mmol/disaccharide unit, weight-average molecular weight 900,000) in water (50 mL)/ethanol (50 mL). The mixture was stirred over night and a 376 mg/5 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 5 hours, the reaction was quenched with acetic acid (107 mg) and sodium chloride (522 mg). Ethanol (250 mL) was added and the resulting precipitation was filtered off and washed twice successively by 80% ethanol, ethanol. The solid was dried in vacuo to afford white solid (475 mg). The introduction ratio of octylamine was 12.6% by HPLC.
- A 30 mg/3 mL solution (ethanol:0.1MHCl=1:1) of octylamine, 1.25 mL of 0.1 M solution (ethanol:water=1:1) of DMT-MM (Wako Pure Chemical Industries, Ltd.) were added to a solution of sodium hyaluronate, (501 mg, 1.25 mmol/disaccharide unit, weight-average molecular weight 900,000) in water (50 mL)/ethanol (50 mL). The mixture was stirred over night and a 381 mg/5 mL aqueous solution of sodium hydrogen carbonate (Japanese Pharmacopoeia) was added. After stirring further 5 hours, the reaction was quenched with acetic acid (107 mg) and sodium chloride (497 mg). Ethanol (250 mL) was added and the resulting precipitation was filtered off and washed twice successively by 80% ethanol, ethanol. The solid was dried in vacuo to afford white solid (497 mg). The introduction ratio of hexadecylamine was 12% by HPLC.
- 64 mg of compound obtained in the above (15-1) was added to 5 mM phosphate buffer saline to give a total amount of 59 ml, then the solution was shaken over night with a shaker. The 0.1% by weight solution of compound prepared in the above (15-1) was obtained.
- Likewise, 0.1% by weight solution of compound prepared in the above (15-2) was obtained.
- The effect of the cinnamate derivative-introduced HA prepared in Example 1 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after peeling the corneal epithelia, 150 μl of saline as the control substance was administered in the left eye, and 150 μl of 0.1% by weight cinnamate derivative-introduced HA solution prepared in the above Example (1-2) as the subject substance was administered in the right eye. At one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hour intervals, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Fourteen model rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.” - The results of the healed area in each individual are shown in
FIG. 17 , and the results of a healing rate in each individual are shown inFIG. 18 . The healed area and the healing rate were calculated as follows. -
Healed area=Area after peeling−Area at each time point (after 1 to 3 days) -
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 17 and 18 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution at all time points ofdays 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% by weight cinnamate derivative-introduced HA solution. - The effect of the octylamine-introduced HA and hexadecylamine-introduced HA prepared in Example 15 on the Migration of rabbit corneal epithelium with surgical removal (The surgical Model).
- The corneal epithelium of the central region was removed by using a trephine (8 mm I.D), a 23 G needle and microscissors after anesthesia with an intravenous injection of 5 mg/kg of ketamine and 2 mg/kg of xylazine and topical administration of 0.4% oxbuprocaine hydrochloride.
- One hour and 4 hours after peeling the corneal epithelia, 150 μl of saline as the control substance was administered in the left eye, and 150 μl of 0.1% octylamine-introduced HA and hexadecylamine-introduced HA solution prepared in the above Example (15-2) as the subject substance was administered in the right eye. At one day and 2 days after the peeling, a total of 4 times with 3 hour intervals, and at 3 days after the peeling, with 3 hour intervals, the same administration as above was performed. In the administration, the 1 ml injection syringes were used. Eight model each rabbits for the corneal epithelial layer disorder described in the above 1) were used as the administration subjects.
- The rabbit was given general anesthesia by intravenously injecting 5 mg/kg of ketamine and 2 mg/kg of xylazine, subsequently, the corneal epithelial loss site was stained with 0.2% sodium fluorescein dissolved in PBS, and photographed under ultra-violet light. The photographing was performed just before the administration of the subject substance one hour after the cornea was peeled and 3 hours after the second administration at 1 to 3 days after the peeling. When photographed, the focal length was made constant to make the magnification of photographs constant.
- The area of the corneal epithelial defective region stained with sodium fluorescein was measured on the printed photograph using the image analyzer. The value obtained by subtracting the area of the peeled
site 3 hours after the final administration at 3 days after the peeling from the area (peeled area) of the peeled site just before the administration of the subject substance one hour after the corneal epithelia were peeled was rendered as “healed area.” - The results of the healed area in each individual are shown in
FIG. 19 , and the results of a healing rate in each individual are shown inFIG. 20 . InFIG. 19 , C8-L(a: control), C8-R(b), C16-L(c: control), C16-R(d) represents left eye to which saline was administrated as a control for C8-R, right eye to which 0.1% octylamine-introduced HA solution was administrated, left eye to which saline was administrated as a control for C16-R, right eye to which 0.1% hexadecylamine-introduced HA solution was administrated, respectively. InFIG. 20 , C8 represents results of the above study using octylamine, and C16 represents results of the above study using hexadecylamine. - The healed area and the healing rate were calculated as follows.
-
Healed area=Area after peeling−Area at each time point (after 1 to 3 days) -
Healing rate=Mean of healed areas at respective time points (after 1 to 3 days) - In
FIGS. 19 and 20 , it was observed that the healed area of the corneal epithelia was significantly increased in the eyes administered with 0.1% octylamine-introduced HA and hexadecylamine-introduced HA solution at all time points ofdays 1 to 3, compared with the healed area of the corneal epithelia in the control eyes. And it was also observed that the healing rate was significantly enhanced in the eyes administered with 0.1% octylamine-introduced HA or hexadecylamine-introduced HA solution.
Claims (36)
1. An agent for applying to mucosa containing glycosaminoglycan into which a hydrophobic group is introduced via a binding chain, as an active ingredient.
2. The agent for applying to mucosa according to claim 1 wherein the glycosaminoglycan is selected from hyaluronic acid, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan sulfate and keratan sulfate, and salts thereof.
3. The agent for applying to mucosa according to claim 1 wherein the glycosaminoglycan is sodium hyaluronate.
4. The agent for applying to mucosa according to claim 1 wherein the hydrophobic group is selected from alkyl groups having 2 to 18 carbon atoms, alkenyl groups having 2 to 18 carbon atoms, alkynyl groups having 2 to 18 carbon atoms, aryl groups, heteroaryl groups, arylalkyl groups, arylalkenyl groups, arylalkynyl groups and amino acid groups.
5. The agent for applying to mucosa according to claim 1 wherein the hydrophobic group is arylalkenyl group.
6. The agent for applying to mucosa according to claim 5 wherein the hydrophobic group is phenylethenyl group.
7. The agent for applying to mucosa according to claim 1 wherein the binding chain is selected from —CONH—, —COO—, -0-, —S03- and —S02NH—.
8. The agent for applying to mucosa according to claim 1 wherein “the glycosaminoglycan into which a hydrophobic group is introduced via a binding chain” further includes a spacer chain between the binding chain and the hydrophobic group (the spacer chain further includes a binding chain at a hydrophobic group side).
9. The agent for applying to mucosa according to claim 8 wherein the spacer chain further having the binding chain at the hydrophobic group side is selected from —COO—(CH2)m—, —COO—(CH2)—(OCH2)n—, —CONH—(CH2)m— and —CONH—(CH2)—(OCH2)n— (wherein m and n represent integers of 1 to 18, respectively).
10. An agent for applying to mucosa containing glycosaminoglycan into which Ph-CH═CH—COO—(CH2)m—NH; Ph-CH═CH—COO—CH2—(OCH2)n—NH; Ph-CH═CH—CONH—(CH2)m—NH; Ph-CH═CH—CONH—CH2—(OCH2)n—NH; Ph-CH═CH—COO—(CH2)m—O; Ph-CH═CH—COO—CH2—(OCH2)n—O; Ph-CH═CH—CONH—(CH2)m—O; Ph-CH═CH—CONH—CH2—(OCH2)n—O; CH3—(CH2)1—COO-Ph-COHN—(CH2)m—NH or CH3—(CH2)1—COO-Ph-CONH—CH2—(OCH2)n—NH (wherein Ph represents phenyl group, m and n represent integers of 1 to 18, respectively, and 1 represents 0 or an integer of 1 to 18) is introduced, as the active ingredient.
11. An agent for applying to mucosa containing glycosaminoglycan having a repeat structure of a structural unit represented by the following formula, as a basic skeleton:
wherein R represents R1 or R2;
Ac represents an acetyl group;
R1 represents ONa or OH;
R2 represents Ph-CH═CH—COO—(CH2)m—NH—; Ph-CH═CH—COO—CH2—(OCH2)n—NH—; Ph-CH═CH—CONH—(CH2)m—NH—; Ph-CH═CH—CONH—CH2—(OCH2)n—NH—; Ph-CH═CH—COO—(CH2)m—O—; Ph-CH═CH—COO—CH2—(OCH2)n—O—; Ph-CH═CH—CONH—(CH2)m—O—; Ph-CH═CH—CONH—CH2—(OCH2)n—O—; CH3(CH2)1—COO-Ph-CONH—(CH2)m—NH—; or CH3(CH2)1—COO-Ph-CONH—CH2—(OCH2)n—NH—; (wherein Ph represents phenyl, m and n represent integers of 1 to 18, respectively, and 1 represents 0 or an integer of 1 to 18) as a basic skeleton, wherein the ratio of the above structural unit wherein R represents R2 is 5 to 30% in molar equivalent relative to the disaccharide repeat unit in molar equivalent of the glycosaminoglycan.
12. The agent for applying to mucosa according to claim 1 wherein the agent is a liquid.
13. The agent for applying to mucosa according to claim 12 wherein the concentration of glycosaminoglycan into which the hydrophobic group is introduced via the binding chain is 0.02 to 5% by weight.
14. The agent for applying to mucosa according to claim 1 wherein the mucosa is cornea, conjunctiva, oral cavity mucosa or urinary bladder mucosa.
15. The agent for applying to mucosa according to claim 1 which is used for the treatment of a mucosal disorder.
16. The agent for applying to mucosa according to claim 15 wherein the mucosal disorder is a mucosal epithelial layer disorder.
17. The agent for applying to mucosa according to claim 1 which is eye drops.
18. The agent for applying to mucosa according to claim 17 which is used for the prevention of a corneal disorder due to ultraviolet rays.
19. A method for applying to mucosa an agent containing glycosaminoglycan into which a hydrophobic group is introduced via a binding chain, as an active ingredient.
20. The method according to claim 19 wherein the glycosaminoglycan is selected from hyaluronic acid, chondroitin, chondroitin sulfate, heparin, heparan sulfate, dermatan sulfate and keratan sulfate, and salts thereof.
21. The method according to claim 19 wherein the glycosaminoglycan is sodium hyaluronate.
22. The method according to claim 19 wherein the hydrophobic group is selected from alkyl groups having 2 to 18 carbon atoms, alkenyl groups having 2 to 18 carbon atoms, alkynyl groups having 2 to 18 carbon atoms, aryl groups, heteroaryl groups, arylalkyl groups, arylalkenyl groups, arylalkynyl groups and amino acid groups.
23. The method according to claim 19 wherein the hydrophobic group is arylalkenyl group.
24. The method according to claim 19 wherein the hydrophobic group is phenylethenyl group.
25. The method according to claim 19 wherein the binding chain is selected from —CONH—, —COO—, -0-, —S03 and —S02NH—.
26. The method according to claim 19 wherein “the glycosaminoglycan into which a hydrophobic group is introduced via a binding chain” further includes a spacer chairs between the binding chain and the hydrophobic group (the spacer chain further includes a binding chain at a hydrophobic group side).
27. The method according to claim 26 wherein the spacer chain further having the binding chain at the hydrophobic group side is selected from —COO—(CH2)m—, —COO—(CH2)—(OCH2)n—, —CONH—(CH2)m— and —CONH—(CH2)—(OCH2)n (wherein m and n represent integers of 1 to 18, respectively).
28. A method for applying to mucosa an agent containing glycosaminoglycan into which Ph-CH═CH—COO—(CH2)m—NH; Ph-CH═CH—COO—CH2—(OCH2)n—NH; Ph-CH═CH—CONH—(CH2)m—NH; Ph-CH═CH—CONH—CH2—(OCH2)n—NH; Ph-CH═CH—COO—(CH2)m—O; Ph-CH═CH—COO—CH2—(OCH2)n—O; Ph-CH═CH—CONH—(CH2)m—O; Ph-CH═CH—CONH—CH2—(OCH2)n—O; CH3—(CH2)1—COO-Ph-COHN—(CH2)m—NH or CH3—(CH2)1—COO-Ph-CONH—CH2—(OCH2)n—NH (wherein Ph represents phenyl group, m and n represent integers of 1 to 18, respectively, and 1 represents 0 or an integer of 1 to 18) is introduced, as an active ingredient.
29. A method for applying to mucosa an agent containing glycosaminoglycan having a repeat structure of a structural unit represented by the following formula, as a basic skeleton:
wherein R represents R1 or R2;
Ac represents an acetyl group;
R1 represents ONa or OH;
R2 represents Ph-CH═CH—COO—(CH2)m—NH—; Ph-CH═CH—COO—CH2—(OCH2)n—NH—; Ph-CH═CH—CONH—(CH2)m—NH—; Ph-CH═CH—CONH—CH2—(OCH2)n—NH—; Ph-CH═CH—COO—(CH2)m—O—; Ph-CH═CH—COO—CH2—(OCH2)n—O—; Ph-CH═CH—CONH—(CH2)m—O—; Ph-CH═CH—CONH—CH2—(OCH2)n—O—; CH3—(CH2)1—COO-Ph-CONH— (CH2)m—NH—; or CH3—(CH2)1COO-Ph-CONH—CH2—(OCH2)n—NH—; (wherein Ph represents phenyl, m and n represent integers of 1 to 18, respectively, and 1 represents 0 or an integer of 1 to 18) as a basic skeleton, wherein the ratio of the above structural unit wherein R represents R2 is 5 to 30% in molar equivalent relative to the disaccharide repeat unit in molar equivalent of the glycosaminoglycan.
30. The method according to claim 19 wherein the agent is a liquid.
31. The method according to claim 30 wherein the concentration of glycosaminoglycan into which the hydrophobic group is introduced via the binding chain is 0.02 to 5% by weight.
32. The method according to claim 19 wherein the mucosa is cornea, conjunctiva, oral cavity mucosa or urinary bladder mucosa.
33. A method for the treatment of a mucosal disorder comprising applying to mucosa an agent containing glycosaminoglycan into which a hydrophobic group is introduced via a binding chain, as an active ingredient.
34. The method according to claim 33 wherein the mucosal disorder is a mucosal epithelial layer disorder.
35. The method according to claim 19 wherein the agent is eye drops.
36. A method for the prevention of a corneal disorder due to ultraviolet rays comprising applying to mucosa eye drops containing glycosaminoglycan into which a hydrophobic group is introduced via a binding chain, as an active ingredient.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2005297170 | 2005-10-12 | ||
JP2005-297170 | 2005-10-12 | ||
JP2006-160478 | 2006-06-09 | ||
JP2006160478 | 2006-06-09 | ||
PCT/JP2006/320801 WO2007043702A1 (en) | 2005-10-12 | 2006-10-12 | Agent for applying to mucosa and method for production thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2006/320801 A-371-Of-International WO2007043702A1 (en) | 2005-10-12 | 2006-10-12 | Agent for applying to mucosa and method for production thereof |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/070,210 Continuation US8969319B2 (en) | 2005-10-12 | 2011-03-23 | Agent for applying to mucosa and method for the production thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
US20080306022A1 true US20080306022A1 (en) | 2008-12-11 |
Family
ID=37942916
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/083,189 Abandoned US20080306022A1 (en) | 2005-10-12 | 2006-10-12 | Agent for Applying to Mucosa and Method for the Production Thereof |
US13/070,210 Active US8969319B2 (en) | 2005-10-12 | 2011-03-23 | Agent for applying to mucosa and method for the production thereof |
US14/600,558 Abandoned US20150196574A1 (en) | 2005-10-12 | 2015-01-20 | Agent for applying to mucosa and method for the production thereof |
US15/941,443 Abandoned US20180289730A1 (en) | 2005-10-12 | 2018-03-30 | Agent for applying to mucosa and method for the production thereof |
Family Applications After (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/070,210 Active US8969319B2 (en) | 2005-10-12 | 2011-03-23 | Agent for applying to mucosa and method for the production thereof |
US14/600,558 Abandoned US20150196574A1 (en) | 2005-10-12 | 2015-01-20 | Agent for applying to mucosa and method for the production thereof |
US15/941,443 Abandoned US20180289730A1 (en) | 2005-10-12 | 2018-03-30 | Agent for applying to mucosa and method for the production thereof |
Country Status (10)
Country | Link |
---|---|
US (4) | US20080306022A1 (en) |
EP (1) | EP1942912B1 (en) |
JP (3) | JP4387445B2 (en) |
KR (4) | KR101411067B1 (en) |
CN (3) | CN103120701B (en) |
AU (1) | AU2006300207C1 (en) |
CA (1) | CA2625532C (en) |
ES (1) | ES2616295T3 (en) |
HK (2) | HK1184700A1 (en) |
WO (1) | WO2007043702A1 (en) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7928089B2 (en) | 2003-09-15 | 2011-04-19 | Vectura Limited | Mucoactive agents for treating a pulmonary disease |
US20110212901A1 (en) * | 2008-11-05 | 2011-09-01 | National University Corporation Tokyo Medical And Dental Unversity | Hyaluronic acid derivative and pharmaceutical composition thereof |
US8329673B2 (en) | 2008-04-04 | 2012-12-11 | University Of Utah Research Foundation | Alkylated semi synthetic glycosaminoglycosan ethers, and methods for making and using thereof |
US9301972B2 (en) | 2006-12-06 | 2016-04-05 | Seikagaku Corporation | Hyaluronic acid derivatives |
US9522162B2 (en) | 2011-03-23 | 2016-12-20 | University Of Utah Research Foundation | Methods for treating or preventing urological inflammation |
US20170348347A1 (en) * | 2014-12-26 | 2017-12-07 | Seikagaku Corporation | Agent for improving ocular subjective symptoms and method thereof |
US20190008889A1 (en) * | 2015-08-20 | 2019-01-10 | Seikagaku Corporation | Agent to be applied to ophthalmic device |
US10525159B2 (en) | 2016-05-26 | 2020-01-07 | Ophtecs Corporation | Liquid preparation for contact lenses comprising hydrolyzed hyaluronic acid derivative and cationic bactericide |
EP2516473B1 (en) | 2009-12-23 | 2020-09-30 | Adocia | Anionic polysaccharides functionalised with a derivative of hydrophobic acid |
US11253540B2 (en) | 2015-05-29 | 2022-02-22 | Seikagaku Corporation | Composition including glycosaminoglycan derivative and chemokine receptor activity regulator |
US11337994B2 (en) | 2016-09-15 | 2022-05-24 | University Of Utah Research Foundation | In situ gelling compositions for the treatment or prevention of inflammation and tissue damage |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103120701B (en) * | 2005-10-12 | 2014-11-05 | 生化学工业株式会社 | Application of gag including hydrophobic group in preparation of drug for preventing or treating epithelium |
JP2009046454A (en) * | 2007-08-22 | 2009-03-05 | Shiseido Co Ltd | Composition for oral cavity |
ITMI20072237A1 (en) * | 2007-11-27 | 2009-05-28 | Sigea Srl | MIXED BUTIRRIC-FORMAL ESTERS OF ACID POLYSACCHARIDES, THEIR PREPARATION AND USE AS DERMOCOSMETICS |
CA2703532C (en) * | 2010-05-10 | 2018-05-01 | Eva Turley | Topically administered, skin-penetrating glycosaminoglycan formulations suitable for use in cosmetic and pharmaceutical applications |
US9067299B2 (en) | 2012-04-25 | 2015-06-30 | Applied Materials, Inc. | Printed chemical mechanical polishing pad |
CN104940221A (en) * | 2015-05-13 | 2015-09-30 | 北京大清生物技术有限公司 | Composition for repairing epithelial injury of bladders and urinary tracts |
EP3871682A1 (en) * | 2016-10-14 | 2021-09-01 | i.com Medical GmbH | Method for establishing, restoring, and preserving homeostasis of the ocular surface |
WO2018175788A1 (en) | 2017-03-22 | 2018-09-27 | Genentech, Inc. | Hydrogel cross-linked hyaluronic acid prodrug compositions and methods |
RU2652581C1 (en) * | 2017-07-03 | 2018-04-26 | федеральное государственное бюджетное образовательное учреждение высшего образования "Приволжский исследовательский медицинский университет" Министерства здравоохранения Российской Федерации | Method of corneoconjunctival xerosis treatment |
US20220370630A1 (en) | 2019-10-03 | 2022-11-24 | Seikagaku Corporation | Transmucosal delivery system for pharmaceutical active ingredient to submucosal tissue of bladder |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5462976A (en) * | 1992-02-05 | 1995-10-31 | Seikagaku Kogyo Kabushiki Kaisha | Photocurable glycosaminoglycan derivatives, crosslinked glycosaminoglycans and method of production thereof |
US5733892A (en) * | 1990-07-24 | 1998-03-31 | Seikagaku Corporation | Metastasis inhibitor composition comprising a phospholipid-linked glycosaminoglycan and method for inhibiting metastasis employing the same |
US6025444A (en) * | 1994-11-17 | 2000-02-15 | Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation) | Cinnamic acid derivative |
US6031017A (en) * | 1995-11-15 | 2000-02-29 | Seikagaku Corporation | Photocured cross-linked-hyaluronic acid gel and method of preparation thereof |
US20010044423A1 (en) * | 1996-12-27 | 2001-11-22 | Katsuya Takahashi | Agent for treatment of bladder troubles |
US6602859B2 (en) * | 2000-12-19 | 2003-08-05 | Seikagaku Corporation | Photocurable hyaluronic acid derivative and process for producing the same, and photocured crosslinked-hyaluronic acid derivative and medical material using the same |
Family Cites Families (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4851521A (en) * | 1985-07-08 | 1989-07-25 | Fidia, S.P.A. | Esters of hyaluronic acid |
JPH0723317B2 (en) * | 1988-03-17 | 1995-03-15 | 生化学工業株式会社 | Corneal epithelial disorder treatment |
DE4203529A1 (en) * | 1992-02-07 | 1993-08-12 | Wolff Walsrode Ag | WATER-SOLUBLE SULFOALKYL HYDROXYALKYL DERIVATIVES OF CELLULOSE AND THEIR USE IN PLASTER AND CEMENT CONTAINERS |
KR960702624A (en) * | 1994-03-14 | 1996-04-27 | 야마타니 와타루 | Material to be worn on the eyeball |
CA2162957C (en) * | 1994-11-17 | 2011-08-02 | Michinori Waki | Cinnamic acid derivative |
JP3308742B2 (en) * | 1994-11-17 | 2002-07-29 | 生化学工業株式会社 | Photocrosslinkable hyaluronic acid derivative, crosslinked product thereof and methods for producing them |
JP3955107B2 (en) * | 1995-05-01 | 2007-08-08 | 生化学工業株式会社 | Method for producing crosslinked polysaccharide |
JP4006039B2 (en) * | 1995-09-13 | 2007-11-14 | 生化学工業株式会社 | Photocrosslinked hyaluronic acid contact lens |
US5789462A (en) * | 1995-09-13 | 1998-08-04 | Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation) | Photocured crosslinked-hyaluronic acid contact lens |
US6232301B1 (en) | 1996-12-27 | 2001-05-15 | Seikagaku Corporation | Remedies for bladder disorders |
JPH1171282A (en) | 1997-09-01 | 1999-03-16 | Senju Pharmaceut Co Ltd | Aqueous preparation for eye drop |
JP3014357B2 (en) | 1998-01-26 | 2000-02-28 | 生化学工業株式会社 | Corneal epithelial layer extension promoter |
ITPD980169A1 (en) * | 1998-07-06 | 2000-01-06 | Fidia Advanced Biopolymers Srl | AMIDES OF HYALURONIC ACID AND ITS DERIVATIVES AND PROCESS FOR THEIR PREPARATION. |
AU3327000A (en) * | 1999-03-24 | 2000-10-09 | Seikagaku Corporation | Artificial saliva |
DE60100866T2 (en) | 2000-04-07 | 2004-07-29 | Laboratoire Medidom S.A. | Ophthalmic medicinal product containing cyclosporine, hyaluronic acid and polysorbate |
JP2001329002A (en) * | 2000-05-25 | 2001-11-27 | Denki Kagaku Kogyo Kk | Modified hyaluronic acid gel, its preparing method and medical material containing same |
JP2002037746A (en) | 2000-07-24 | 2002-02-06 | Fuji Yakuhin:Kk | Liquid preparation containing quinolonecarboxylic acid based antimicrobial agent |
JP4172176B2 (en) | 2000-12-19 | 2008-10-29 | 生化学工業株式会社 | Photoreactive hyaluronic acid, process for producing the same, photocrosslinked hyaluronic acid and medical material |
AU2002243630A1 (en) * | 2001-01-23 | 2002-08-06 | Massachusetts Institute Of Technology | Solid- and solution -phase synthesis of heparin and other glycosaminoglycans |
JP4273208B2 (en) * | 2001-03-30 | 2009-06-03 | チッソ株式会社 | Gynecological disease treatment |
EP2287205A3 (en) * | 2004-09-15 | 2012-08-01 | Seikagaku Corporation | Photoreactive polysaccharide. photocrosslinked polysaccharide products, the method of making them and medical materials made therefrom |
CN103120701B (en) * | 2005-10-12 | 2014-11-05 | 生化学工业株式会社 | Application of gag including hydrophobic group in preparation of drug for preventing or treating epithelium |
JP7023317B2 (en) | 2020-05-12 | 2022-02-21 | 沖電気工業株式会社 | Optical wavelength filter and wavelength separation optical circuit |
-
2006
- 2006-10-12 CN CN201210562380.9A patent/CN103120701B/en not_active Expired - Fee Related
- 2006-10-12 CN CN201210562963.1A patent/CN103120702B/en not_active Expired - Fee Related
- 2006-10-12 KR KR1020087011277A patent/KR101411067B1/en active IP Right Grant
- 2006-10-12 KR KR1020137024561A patent/KR20130109259A/en not_active Application Discontinuation
- 2006-10-12 ES ES06811990.8T patent/ES2616295T3/en active Active
- 2006-10-12 CA CA2625532A patent/CA2625532C/en active Active
- 2006-10-12 WO PCT/JP2006/320801 patent/WO2007043702A1/en active Application Filing
- 2006-10-12 KR KR1020147008031A patent/KR101554834B1/en active IP Right Grant
- 2006-10-12 US US12/083,189 patent/US20080306022A1/en not_active Abandoned
- 2006-10-12 KR KR1020117023949A patent/KR101428153B1/en active IP Right Grant
- 2006-10-12 AU AU2006300207A patent/AU2006300207C1/en not_active Ceased
- 2006-10-12 JP JP2008519554A patent/JP4387445B2/en active Active
- 2006-10-12 CN CN2006800380199A patent/CN101287473B/en active Active
- 2006-10-12 EP EP06811990.8A patent/EP1942912B1/en active Active
-
2009
- 2009-08-11 JP JP2009186548A patent/JP5094799B2/en not_active Expired - Fee Related
-
2011
- 2011-03-23 US US13/070,210 patent/US8969319B2/en active Active
-
2012
- 2012-06-27 JP JP2012144630A patent/JP2012211171A/en active Pending
-
2013
- 2013-10-31 HK HK13112267.6A patent/HK1184700A1/en not_active IP Right Cessation
- 2013-10-31 HK HK13112270.1A patent/HK1184701A1/en not_active IP Right Cessation
-
2015
- 2015-01-20 US US14/600,558 patent/US20150196574A1/en not_active Abandoned
-
2018
- 2018-03-30 US US15/941,443 patent/US20180289730A1/en not_active Abandoned
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5733892A (en) * | 1990-07-24 | 1998-03-31 | Seikagaku Corporation | Metastasis inhibitor composition comprising a phospholipid-linked glycosaminoglycan and method for inhibiting metastasis employing the same |
US5462976A (en) * | 1992-02-05 | 1995-10-31 | Seikagaku Kogyo Kabushiki Kaisha | Photocurable glycosaminoglycan derivatives, crosslinked glycosaminoglycans and method of production thereof |
US6025444A (en) * | 1994-11-17 | 2000-02-15 | Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation) | Cinnamic acid derivative |
US6031017A (en) * | 1995-11-15 | 2000-02-29 | Seikagaku Corporation | Photocured cross-linked-hyaluronic acid gel and method of preparation thereof |
US20010044423A1 (en) * | 1996-12-27 | 2001-11-22 | Katsuya Takahashi | Agent for treatment of bladder troubles |
US6602859B2 (en) * | 2000-12-19 | 2003-08-05 | Seikagaku Corporation | Photocurable hyaluronic acid derivative and process for producing the same, and photocured crosslinked-hyaluronic acid derivative and medical material using the same |
Cited By (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7928089B2 (en) | 2003-09-15 | 2011-04-19 | Vectura Limited | Mucoactive agents for treating a pulmonary disease |
US9616087B2 (en) | 2006-12-06 | 2017-04-11 | Seikagaku Corporation | Pharmaceutical agent having long-lasting effect of treating arthritic disorders |
US9301972B2 (en) | 2006-12-06 | 2016-04-05 | Seikagaku Corporation | Hyaluronic acid derivatives |
US8329673B2 (en) | 2008-04-04 | 2012-12-11 | University Of Utah Research Foundation | Alkylated semi synthetic glycosaminoglycosan ethers, and methods for making and using thereof |
US9549945B2 (en) | 2008-04-04 | 2017-01-24 | University Of Utah Research Foundation | Use of alkylated semi-synthetic glycosaminoglycosan ethers for the treatment of inflammation |
US20110212901A1 (en) * | 2008-11-05 | 2011-09-01 | National University Corporation Tokyo Medical And Dental Unversity | Hyaluronic acid derivative and pharmaceutical composition thereof |
US8759322B2 (en) | 2008-11-05 | 2014-06-24 | National University Corporation Tokyo Medical And Dental University | Hyaluronic acid derivative and pharmaceutical composition thereof |
EP2516473B1 (en) | 2009-12-23 | 2020-09-30 | Adocia | Anionic polysaccharides functionalised with a derivative of hydrophobic acid |
US10226481B2 (en) | 2011-03-23 | 2019-03-12 | University Of Utah Research Foundation | Pharmaceutical compositions composed of low molecular weight sulfated hyaluronan |
US9522162B2 (en) | 2011-03-23 | 2016-12-20 | University Of Utah Research Foundation | Methods for treating or preventing urological inflammation |
US20170348347A1 (en) * | 2014-12-26 | 2017-12-07 | Seikagaku Corporation | Agent for improving ocular subjective symptoms and method thereof |
US11147831B2 (en) | 2014-12-26 | 2021-10-19 | Seikagaku Corporation | Agent for improving ocular subjective symptoms and method thereof |
US11185558B2 (en) | 2014-12-26 | 2021-11-30 | Seikagaku Corporation | Agent for improving ocular subjective symptoms and method thereof |
US11779593B2 (en) | 2014-12-26 | 2023-10-10 | Seikagaku Corporation | Agent for improving ocular subjective symptoms and method thereof |
US11253540B2 (en) | 2015-05-29 | 2022-02-22 | Seikagaku Corporation | Composition including glycosaminoglycan derivative and chemokine receptor activity regulator |
US20190008889A1 (en) * | 2015-08-20 | 2019-01-10 | Seikagaku Corporation | Agent to be applied to ophthalmic device |
US10525159B2 (en) | 2016-05-26 | 2020-01-07 | Ophtecs Corporation | Liquid preparation for contact lenses comprising hydrolyzed hyaluronic acid derivative and cationic bactericide |
US11337994B2 (en) | 2016-09-15 | 2022-05-24 | University Of Utah Research Foundation | In situ gelling compositions for the treatment or prevention of inflammation and tissue damage |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20180289730A1 (en) | Agent for applying to mucosa and method for the production thereof | |
ES2665254T3 (en) | Compositions comprising hyaluronic acid for treatment and prevention of mucosal related diseases | |
ES2565641T3 (en) | Sulfated polysaccharide compound and preparation and use thereof | |
JPH09169667A (en) | New medicine composition | |
JP2009511423A5 (en) | ||
CN111848830A (en) | Application of fluorine-containing compound modified chitosan as drug carrier and preparation method thereof | |
CN1694727A (en) | Injections for eye tissue containing drug bound to polyethylene glycol | |
WO2001074400A1 (en) | Transporters and drug delivery system by using the same | |
ES2905606T3 (en) | Hyaluronan conjugates with pharmaceutically active substances, methods and compositions | |
ES2869943T3 (en) | Transmembrane Drug Delivery System | |
AU2012201314B2 (en) | Agent for applying to mucosa and method for production thereof | |
US20220133776A1 (en) | Composition including glucosaminoglycan derivative and chemokine receptor activity regulator | |
WO2018201007A2 (en) | Dendrimer compositions for use in angiography | |
WO2024067874A1 (en) | Modified chitosan and use thereof in large molecule active ingredient delivery |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: SEIKAGAKU CORPORATION, JAPAN Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MIYAMOTO, KENJI;TAKAHASHI, KATSUYA;SHIMOJIMA, YUUJI;REEL/FRAME:020852/0626 Effective date: 20080328 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |