CA2482320C - Methods for modulating phototoxicity - Google Patents
Methods for modulating phototoxicity Download PDFInfo
- Publication number
- CA2482320C CA2482320C CA2482320A CA2482320A CA2482320C CA 2482320 C CA2482320 C CA 2482320C CA 2482320 A CA2482320 A CA 2482320A CA 2482320 A CA2482320 A CA 2482320A CA 2482320 C CA2482320 C CA 2482320C
- Authority
- CA
- Canada
- Prior art keywords
- molecules
- light
- proline
- cells
- photodamage
- 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.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title abstract description 8
- 206010034972 Photosensitivity reaction Diseases 0.000 title description 7
- 208000007578 phototoxic dermatitis Diseases 0.000 title description 6
- 231100000018 phototoxicity Toxicity 0.000 title description 6
- 230000008832 photodamage Effects 0.000 claims abstract description 17
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 10
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims description 7
- 125000004432 carbon atom Chemical group C* 0.000 claims description 5
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 claims description 4
- 125000000217 alkyl group Chemical group 0.000 claims description 4
- 229960002591 hydroxyproline Drugs 0.000 claims description 4
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 claims description 3
- 125000005907 alkyl ester group Chemical group 0.000 claims description 2
- 239000006210 lotion Substances 0.000 claims description 2
- 125000001500 prolyl group Chemical group [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 claims description 2
- 239000002453 shampoo Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims 1
- 108010035532 Collagen Proteins 0.000 abstract description 6
- 102000008186 Collagen Human genes 0.000 abstract description 6
- 229920001436 collagen Polymers 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 6
- 150000001875 compounds Chemical class 0.000 description 33
- 210000004027 cell Anatomy 0.000 description 28
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 18
- GRFNBEZIAWKNCO-UHFFFAOYSA-N 3-pyridinol Chemical compound OC1=CC=CN=C1 GRFNBEZIAWKNCO-UHFFFAOYSA-N 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 14
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 9
- 238000010791 quenching Methods 0.000 description 9
- 230000000171 quenching effect Effects 0.000 description 9
- 229940011671 vitamin b6 Drugs 0.000 description 9
- 238000003556 assay Methods 0.000 description 8
- 230000004663 cell proliferation Effects 0.000 description 8
- 239000011726 vitamin B6 Substances 0.000 description 8
- 235000019158 vitamin B6 Nutrition 0.000 description 8
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 7
- LCYXYLLJXMAEMT-SAXRGWBVSA-N Pyridinoline Chemical compound OC(=O)[C@@H](N)CCC1=C[N+](C[C@H](O)CC[C@H](N)C([O-])=O)=CC(O)=C1C[C@H](N)C(O)=O LCYXYLLJXMAEMT-SAXRGWBVSA-N 0.000 description 7
- 229940098773 bovine serum albumin Drugs 0.000 description 7
- GDOPTJXRTPNYNR-UHFFFAOYSA-N methyl-cyclopentane Natural products CC1CCCC1 GDOPTJXRTPNYNR-UHFFFAOYSA-N 0.000 description 7
- 230000035755 proliferation Effects 0.000 description 7
- 229960002429 proline Drugs 0.000 description 7
- 210000003491 skin Anatomy 0.000 description 7
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 150000003147 proline derivatives Chemical class 0.000 description 5
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 210000002510 keratinocyte Anatomy 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 231100000489 sensitizer Toxicity 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 206010006187 Breast cancer Diseases 0.000 description 3
- 208000026310 Breast neoplasm Diseases 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 230000005281 excited state Effects 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- -1 i.e. Chemical compound 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 231100000760 phototoxic Toxicity 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 210000004927 skin cell Anatomy 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 208000002874 Acne Vulgaris Diseases 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- 201000004681 Psoriasis Diseases 0.000 description 2
- 206010070834 Sensitisation Diseases 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 206010000496 acne Diseases 0.000 description 2
- 208000009621 actinic keratosis Diseases 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001028 anti-proliverative effect Effects 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 230000005779 cell damage Effects 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 description 2
- 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 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 230000005283 ground state Effects 0.000 description 2
- 230000003463 hyperproliferative effect Effects 0.000 description 2
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 150000002978 peroxides Chemical class 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000008313 sensitization Effects 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000002626 targeted therapy Methods 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- IMNIMPAHZVJRPE-UHFFFAOYSA-N triethylenediamine Chemical compound C1CN2CCN1CC2 IMNIMPAHZVJRPE-UHFFFAOYSA-N 0.000 description 2
- RKMGAJGJIURJSJ-UHFFFAOYSA-N 2,2,6,6-tetramethylpiperidine Chemical compound CC1(C)CCCC(C)(C)N1 RKMGAJGJIURJSJ-UHFFFAOYSA-N 0.000 description 1
- 125000004042 4-aminobutyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])N([H])[H] 0.000 description 1
- GCNTZFIIOFTKIY-UHFFFAOYSA-N 4-hydroxypyridine Chemical compound OC1=CC=NC=C1 GCNTZFIIOFTKIY-UHFFFAOYSA-N 0.000 description 1
- 108010005094 Advanced Glycation End Products Proteins 0.000 description 1
- 108090000672 Annexin A5 Proteins 0.000 description 1
- 102000004121 Annexin A5 Human genes 0.000 description 1
- 102100031109 Beta-catenin-like protein 1 Human genes 0.000 description 1
- 101710164563 Beta-catenin-like protein 1 Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- VVNCNSJFMMFHPL-VKHMYHEASA-N D-penicillamine Chemical compound CC(C)(S)[C@@H](N)C(O)=O VVNCNSJFMMFHPL-VKHMYHEASA-N 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 230000007018 DNA scission Effects 0.000 description 1
- 238000004435 EPR spectroscopy Methods 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010051246 Photodermatosis Diseases 0.000 description 1
- 239000002262 Schiff base Substances 0.000 description 1
- 150000004753 Schiff bases Chemical class 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 108091092356 cellular DNA Proteins 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 238000000326 densiometry Methods 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 239000012973 diazabicyclooctane Substances 0.000 description 1
- 231100000676 disease causative agent Toxicity 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 229960002885 histidine Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 150000004806 hydroxypyridines Chemical class 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 230000035407 negative regulation of cell proliferation Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 229960001639 penicillamine Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000008845 photoaging Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- UBQKCCHYAOITMY-UHFFFAOYSA-N pyridin-2-ol Chemical compound OC1=CC=CC=N1 UBQKCCHYAOITMY-UHFFFAOYSA-N 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 201000008261 skin carcinoma Diseases 0.000 description 1
- 210000001626 skin fibroblast Anatomy 0.000 description 1
- 231100000051 skin sensitiser Toxicity 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- MFPWEWYKQYMWRO-UHFFFAOYSA-N tert-butyl carboxy carbonate Chemical compound CC(C)(C)OC(=O)OC(O)=O MFPWEWYKQYMWRO-UHFFFAOYSA-N 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 229950003937 tolonium Drugs 0.000 description 1
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/004—Aftersun preparations
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/401—Proline; Derivatives thereof, e.g. captopril
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
- A61K8/4913—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having five membered rings, e.g. pyrrolidone carboxylic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
- A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
-
- 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
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
-
- 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
- A61P17/16—Emollients or protectives, e.g. against radiation
-
- 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
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/57—Compounds covalently linked to a(n inert) carrier molecule, e.g. conjugates, pro-fragrances
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Birds (AREA)
- Toxicology (AREA)
- Ophthalmology & Optometry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Pyrrole Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
- Electrochromic Elements, Electrophoresis, Or Variable Reflection Or Absorption Elements (AREA)
- Radiation-Therapy Devices (AREA)
- Photovoltaic Devices (AREA)
- Pyridine Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to methods for modulating photodamage via the use of collagen derived molecules which either enhance or inhibit damage caused by ultraviolet light.
Description
METHODS FOR IYIODULATING PHOTOTOXICITY
FIELD OF THE INVENTION
This invention relates to molecules and compositions whieli are useful as modulators of skin cell phototoxicity. Specifically, "modulators" as used herein refers to materials which can either accelerate or retard the damage of cells, such as skin cells, caused by exposure to light.
BACKGROUND AND PRIOR ART
It is well Icnown that light, UVA light in particular, is dan-iaging to skin cells.
Phototoxic cell damage occurs via the reaction of liglzt with certain coinpounds that are eia.dogenous to skin. The mechanism by which the photodamage occurs is well understood, and can be described, briefly, as follows. The molecules involved, which may be referred to as sensitizers or even accelerators of slcin dainage, react with light and, in the presence of oxygen, result in the fonilation of "reactive oxygen species", or "ROS." It is these molecules, i.e., ROS molecules, which are involved in pathways leading to cell damage, ineludiulg carcinogenesis and photoaging, but not being limited to these phenomena. More details of this phenomenon may be found in Wondrak, et al. J. Invest. DemZatol 119:489-498 (2002).
The fact that molecules endogenous to, e.g., the skin, are involved in accelerated pliototoxicity suggests targeted therapy. To elaborate, if a compound is essentially irterk in the absence of light but is involved in cellular destruction upon contact with light, then such compounds could be used in situations where targeted cell deatli is desired.
Such situations include, but are not limited to, psoriasis, acne, premalignant and malignant hyperproliferative disorders such as actinic keratosis, and other conditions well laiown to the art.
Conversely, the existence of the photoactivable molecules suggests the existence of molecules which act to quench or to irihibit the effect of ligllt on cells.
Sucli quenchers or inhibitors can be used in situations where the harmful effects of light need to be reversed, and/or inhibited. Such quenchers or inhibitors may be used propliylactically, as well as therapeutically.
Hence, the modulation of phototoxicity on cells is the focus of the invention described herein, as will be seen in the disclosure which follows. Modulators, as used herein, refers to molecules which may be derived from slcin components, collagen in particular.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 sets forth the structures of various molecules employed in the examples.
Figure 2 depicts results showing that molecules with a 3 hydroxypyridine central structure were efficacious in provoking inhibition of cell proliferation, when exposed to light.
Figure 3 coinpares the results obtained for the 3-hydroxpyridine compounds of figure 1, on HaCaT cells, which are keratinocytes, in the presence of light.
Figure 4 depicts result obtained on malignant melanoma, using the N-ethyl derivative described herein.
Figure 5 parallels figure 4, but the cells used were breast cancer cells.
Figure 6 presents, via FACS (flow cytometric) analysis, evidence that the N-ethyl derivative pushes cells into apoptosis.
Figure 7 presents schematically the synthesis of BSA-B6 complexes.
Figure 8 shows that the coinplexes of figure 7 were effective in inhibiting cell proliferation.
Figure 9 is a proposed mechanism for the quenching of photoactivated molecules by energy transfer ("ET" in the figure). "S*" is the fully excited sensitizer, while "S" is the sensitizer.
Figure 10 sets forth structures of proline derivatives tested as quenchers.
Figure 11 depicts results from assays designed to show the efficacy of the proline derivatives in quenching photosensitized, DNA damage.
Figure 12 shows how it was determined that compounds described herein quenched
FIELD OF THE INVENTION
This invention relates to molecules and compositions whieli are useful as modulators of skin cell phototoxicity. Specifically, "modulators" as used herein refers to materials which can either accelerate or retard the damage of cells, such as skin cells, caused by exposure to light.
BACKGROUND AND PRIOR ART
It is well Icnown that light, UVA light in particular, is dan-iaging to skin cells.
Phototoxic cell damage occurs via the reaction of liglzt with certain coinpounds that are eia.dogenous to skin. The mechanism by which the photodamage occurs is well understood, and can be described, briefly, as follows. The molecules involved, which may be referred to as sensitizers or even accelerators of slcin dainage, react with light and, in the presence of oxygen, result in the fonilation of "reactive oxygen species", or "ROS." It is these molecules, i.e., ROS molecules, which are involved in pathways leading to cell damage, ineludiulg carcinogenesis and photoaging, but not being limited to these phenomena. More details of this phenomenon may be found in Wondrak, et al. J. Invest. DemZatol 119:489-498 (2002).
The fact that molecules endogenous to, e.g., the skin, are involved in accelerated pliototoxicity suggests targeted therapy. To elaborate, if a compound is essentially irterk in the absence of light but is involved in cellular destruction upon contact with light, then such compounds could be used in situations where targeted cell deatli is desired.
Such situations include, but are not limited to, psoriasis, acne, premalignant and malignant hyperproliferative disorders such as actinic keratosis, and other conditions well laiown to the art.
Conversely, the existence of the photoactivable molecules suggests the existence of molecules which act to quench or to irihibit the effect of ligllt on cells.
Sucli quenchers or inhibitors can be used in situations where the harmful effects of light need to be reversed, and/or inhibited. Such quenchers or inhibitors may be used propliylactically, as well as therapeutically.
Hence, the modulation of phototoxicity on cells is the focus of the invention described herein, as will be seen in the disclosure which follows. Modulators, as used herein, refers to molecules which may be derived from slcin components, collagen in particular.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 sets forth the structures of various molecules employed in the examples.
Figure 2 depicts results showing that molecules with a 3 hydroxypyridine central structure were efficacious in provoking inhibition of cell proliferation, when exposed to light.
Figure 3 coinpares the results obtained for the 3-hydroxpyridine compounds of figure 1, on HaCaT cells, which are keratinocytes, in the presence of light.
Figure 4 depicts result obtained on malignant melanoma, using the N-ethyl derivative described herein.
Figure 5 parallels figure 4, but the cells used were breast cancer cells.
Figure 6 presents, via FACS (flow cytometric) analysis, evidence that the N-ethyl derivative pushes cells into apoptosis.
Figure 7 presents schematically the synthesis of BSA-B6 complexes.
Figure 8 shows that the coinplexes of figure 7 were effective in inhibiting cell proliferation.
Figure 9 is a proposed mechanism for the quenching of photoactivated molecules by energy transfer ("ET" in the figure). "S*" is the fully excited sensitizer, while "S" is the sensitizer.
Figure 10 sets forth structures of proline derivatives tested as quenchers.
Figure 11 depicts results from assays designed to show the efficacy of the proline derivatives in quenching photosensitized, DNA damage.
Figure 12 shows how it was determined that compounds described herein quenched
-2-singlet oxygen.
Figure 13 shows the protective effect of various quencher molecules which inhibit skizl cell photodamage.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention relates to use of a photodamage modulator for enhancing photodamage caused by ultraviolet light in a subject in need thereof, wherein the modulator is :
^ a modulator which has a structure consisting of an N-alkyl-3-hydroxypyridine or salt thereof, wherein the alkyl moiety is 2-20 carbon atoms, ^ vitamin B6, ^ pyridinoline, or ^ 3-hydroxypyridine.
The present invention also relates to use of a photodamage modulator for treating photodamage caused by ultraviolet light in a human subject in need thereof, wherein the modulator is proline, 4-hydroxyproline or an alkyl ester thereof.
Figure 13 shows the protective effect of various quencher molecules which inhibit skizl cell photodamage.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention relates to use of a photodamage modulator for enhancing photodamage caused by ultraviolet light in a subject in need thereof, wherein the modulator is :
^ a modulator which has a structure consisting of an N-alkyl-3-hydroxypyridine or salt thereof, wherein the alkyl moiety is 2-20 carbon atoms, ^ vitamin B6, ^ pyridinoline, or ^ 3-hydroxypyridine.
The present invention also relates to use of a photodamage modulator for treating photodamage caused by ultraviolet light in a human subject in need thereof, wherein the modulator is proline, 4-hydroxyproline or an alkyl ester thereof.
-3-EXAMPLE I
Components of the endogenous skin component, to collagen were believed to be most likely the causative agents of phototoxic damage and/or iiAzibition thereof.
Hence, such molecules were investigated.
Pyridinoline is an amino acid involved in cross linlcing collagen molecules, and it was ulvestigated for its role in phototoxicity. The structure of pyridinoline is well known, and is depicted in figure 1, together with the structure of other molecules described herein.
A series of experinlents were can-ied out on HaCaT keratinocytes, and human fibroblasts. hi these experiments, cell samples were contacted with 500 M of pyridinoline, 500 M of desinosine, wliich is an elastin constituent with a structure related to pyridinoline, or 500 M of vitamin B6 (pyridoxine). Controls received no added compound. In one set of experiments, tlie cells received no external light source. In another set of experiments, they received WA light, at 3.3 Jlcm2. The CF3 fibroblasts received solar simulated light, or "SSL," which combines 2.3 J/cm2 of LNA light, and 0.12 J/cm` of UVB light.
Results are shown, in figure 2, in temis of percent of cell proliferation relative to a control (no added compound, no light). The measurements were taken 3 days after stimulation.
The results indicated that pyridinoline had an antiproliferative effect, but oiily in the presence of light. Vitamin B6 showed dramatically more efficacy in. inhibiting the cellular proliferation.
A second set of experiments were carried out in which catalase, which is a peroxide scavenger, was added, at 400 /ml. The catalase had absolutely no effect on vitamin B6 sensitization, suggesting that a mechanism other than peroxide formation was involved for this molecule.
Based upon these results, structures of the compounds were conxpared to determine if -3a-a common structural feature of the molecules, or a"pharmacopnore" could be identified. It was noted that botll vitamin B6 and pyridinoline share a 3-hydroxypyridine central structure, which in turn suggested the next series of experiments.
A group of hydroxypyridine derivatives were studied, in experiments paralleling those described su ra. In brief, 2, 3, and 4-hydroxypyridine were tested, as was N-ethyl-3-liydroxypyridine. All structures are set forth in figure 1.
HaCaT cells, as described supra, were tested in a proliferation inhibition assay. Cell samples received equal amounts of one of the 4 coinpounds listed su ra and were contacted with or without either solar siinulated light ("SSL") as described, supra, or WA light alone, also as described supra.
The results are depicted in figure 3.
As coinpared to controls, 3-hydroxypyridine had an inhibitory effect, where the N-ethyl derivative had a killing effect. The N-ethyl derivative also functioned in the presence of both UVA and UVB light, whereas 3-hydroxypyridine functioned in the presence of UVB
light as a cell proliferation inhibitor.
The experiments described supra were followed by additional experiments using malignant melanoma cells (G-361 cells ), and malignant breast cancer cells (MCF-7). In these experiments, the N-ethyl derivative described supra was tested as described, at varying concentrations, with UVA light at 9.9 J/cm2. Viability was measured two days following the treatment. As controls, experiments were ran using only the N-ethyl derivative, and only the UVA light.
The results, shown in figures 4 (G-361 cells) and 5 (MCF7 cells), show that the combination led to pronounced cytotoxicity.
When the G-361 cells were analyzed via FACS, the data showed that the cells were driven into programined cell death, i.e., apoptosis. This is seen in figure 6, where staining with the apoptic markers annexin V and propidium iodide showed a sharp increase in stained
Components of the endogenous skin component, to collagen were believed to be most likely the causative agents of phototoxic damage and/or iiAzibition thereof.
Hence, such molecules were investigated.
Pyridinoline is an amino acid involved in cross linlcing collagen molecules, and it was ulvestigated for its role in phototoxicity. The structure of pyridinoline is well known, and is depicted in figure 1, together with the structure of other molecules described herein.
A series of experinlents were can-ied out on HaCaT keratinocytes, and human fibroblasts. hi these experiments, cell samples were contacted with 500 M of pyridinoline, 500 M of desinosine, wliich is an elastin constituent with a structure related to pyridinoline, or 500 M of vitamin B6 (pyridoxine). Controls received no added compound. In one set of experiments, tlie cells received no external light source. In another set of experiments, they received WA light, at 3.3 Jlcm2. The CF3 fibroblasts received solar simulated light, or "SSL," which combines 2.3 J/cm2 of LNA light, and 0.12 J/cm` of UVB light.
Results are shown, in figure 2, in temis of percent of cell proliferation relative to a control (no added compound, no light). The measurements were taken 3 days after stimulation.
The results indicated that pyridinoline had an antiproliferative effect, but oiily in the presence of light. Vitamin B6 showed dramatically more efficacy in. inhibiting the cellular proliferation.
A second set of experiments were carried out in which catalase, which is a peroxide scavenger, was added, at 400 /ml. The catalase had absolutely no effect on vitamin B6 sensitization, suggesting that a mechanism other than peroxide formation was involved for this molecule.
Based upon these results, structures of the compounds were conxpared to determine if -3a-a common structural feature of the molecules, or a"pharmacopnore" could be identified. It was noted that botll vitamin B6 and pyridinoline share a 3-hydroxypyridine central structure, which in turn suggested the next series of experiments.
A group of hydroxypyridine derivatives were studied, in experiments paralleling those described su ra. In brief, 2, 3, and 4-hydroxypyridine were tested, as was N-ethyl-3-liydroxypyridine. All structures are set forth in figure 1.
HaCaT cells, as described supra, were tested in a proliferation inhibition assay. Cell samples received equal amounts of one of the 4 coinpounds listed su ra and were contacted with or without either solar siinulated light ("SSL") as described, supra, or WA light alone, also as described supra.
The results are depicted in figure 3.
As coinpared to controls, 3-hydroxypyridine had an inhibitory effect, where the N-ethyl derivative had a killing effect. The N-ethyl derivative also functioned in the presence of both UVA and UVB light, whereas 3-hydroxypyridine functioned in the presence of UVB
light as a cell proliferation inhibitor.
The experiments described supra were followed by additional experiments using malignant melanoma cells (G-361 cells ), and malignant breast cancer cells (MCF-7). In these experiments, the N-ethyl derivative described supra was tested as described, at varying concentrations, with UVA light at 9.9 J/cm2. Viability was measured two days following the treatment. As controls, experiments were ran using only the N-ethyl derivative, and only the UVA light.
The results, shown in figures 4 (G-361 cells) and 5 (MCF7 cells), show that the combination led to pronounced cytotoxicity.
When the G-361 cells were analyzed via FACS, the data showed that the cells were driven into programined cell death, i.e., apoptosis. This is seen in figure 6, where staining with the apoptic markers annexin V and propidium iodide showed a sharp increase in stained
-4-cells when the N-ethyl derivative was used, especially with U VA at a concentration of 3.3 J/ma.
The effective materials of the invention are small molecules. While small molecules are useful, it is sometimes desirable to complex such molecules to larger molecules, such as proteins. This facilitates targeting of the small molecule if it is complexed with, e.g., an antibody specific for a particular marlcer on cells, a ligand for a particular receptor, a nuclear associated protein and so forth.
To test the feasibility of this approach, vitamin B6 molecules were coupled to bovine serum albumin. In brief, a lysine side chain of the bovine serum albumin (BSA) molecule was covalently modified by reacting 350 mg of it with vitamin B6, which is a pyridoxal aldehyde (64 mg), to form a Schiff base. In turn, the Scliiff base was reduced with NaCNBH3 (58 mg) in 1.5 ml of 0.25 M phosphate buffer (pH 7.4), overnight at 37 C, and dialyzed extensively (48 hours, 4 C). The resulting BSA-B6 adduct was characterized by mass spectrometry and fluorescence spectroscopy. The protein was then lyophilized, and used in the examples which follow. Figure 7 depicts the synthesis. The spectroscopy worlc indicated that, on average, each BSA molecule was complexed to 5-6 pyroxidal molecules.
The antiproliferative effect of the complexes described in example 4 were tested, by adding either nothing (control), BSA, or the BSA-B6 complexes, and treating or not treating samples of HaCaT keratinocytes with SSL. The BSA and BSA-B6 were added at 10 mg/ml, and the SSL was 2.3 J/cm2 of UVA plus 0.12 J/cma UVB. Three days after treatment, proliferation was measured using a Coulter counter, and standard methods.
It will be seen from the results shown in figure 8 that the BSA-B6 complexes were very effective in inhibiting the proliferation of the keratinocytes.
The data set forth in examples 1-5, su ra, deal with molecules which enhance cellular destruction. Such is not always desirable, llowever, and in this example and those which follow, experiments are set forth which describe molecules which inhibit this process. These
The effective materials of the invention are small molecules. While small molecules are useful, it is sometimes desirable to complex such molecules to larger molecules, such as proteins. This facilitates targeting of the small molecule if it is complexed with, e.g., an antibody specific for a particular marlcer on cells, a ligand for a particular receptor, a nuclear associated protein and so forth.
To test the feasibility of this approach, vitamin B6 molecules were coupled to bovine serum albumin. In brief, a lysine side chain of the bovine serum albumin (BSA) molecule was covalently modified by reacting 350 mg of it with vitamin B6, which is a pyridoxal aldehyde (64 mg), to form a Schiff base. In turn, the Scliiff base was reduced with NaCNBH3 (58 mg) in 1.5 ml of 0.25 M phosphate buffer (pH 7.4), overnight at 37 C, and dialyzed extensively (48 hours, 4 C). The resulting BSA-B6 adduct was characterized by mass spectrometry and fluorescence spectroscopy. The protein was then lyophilized, and used in the examples which follow. Figure 7 depicts the synthesis. The spectroscopy worlc indicated that, on average, each BSA molecule was complexed to 5-6 pyroxidal molecules.
The antiproliferative effect of the complexes described in example 4 were tested, by adding either nothing (control), BSA, or the BSA-B6 complexes, and treating or not treating samples of HaCaT keratinocytes with SSL. The BSA and BSA-B6 were added at 10 mg/ml, and the SSL was 2.3 J/cm2 of UVA plus 0.12 J/cma UVB. Three days after treatment, proliferation was measured using a Coulter counter, and standard methods.
It will be seen from the results shown in figure 8 that the BSA-B6 complexes were very effective in inhibiting the proliferation of the keratinocytes.
The data set forth in examples 1-5, su ra, deal with molecules which enhance cellular destruction. Such is not always desirable, llowever, and in this example and those which follow, experiments are set forth which describe molecules which inhibit this process. These
-5-molecules will be referred to as quenchers of photoexcited states, or "QPES"
hereafter. Such compounds are characterized by an ability to inactivate the photoexcited state of a rinolecule which would then provoke the type of cell death described supra.
The proposed mechanism by which these molecules function is set forth in figure 9, although it is to be noted that applicants do not wish to be bound by this proposed mechanism. In brief, UV irradiation of a molecule leads to excitation of electrons (excited states, as "S*" in the figure), together with fonnation of excited singlets, and, after intersystem crossing (ISC), triplet states. These are key intermediates in the photodamage of cells. QPES compounds nullify this effect by accepting the excitation energy of the compounds associated witll photodamage by energy transfer ("ET"), neutralizing phototoxic intennediates which relax back to ground state, dissipating the energy via hannless vibrational energy, or heat. The QPES conipotulds themselves are not depleted in this process, and neutralize multiple photodainaging molecules.
It is well lulown that q>X174 plasmid is cleaved only by combined action of irradiation from solar sin-iulated liglit, and AGE-pigment enriclied protein, which acts as a UV sensitizer. AGE-BSA ("advanced glycation end product" modified bovine serum albumin), is a model for accumulation of endogenous skin sensitizers of the type described suUra. Details of the assay showing this will be found in Wondrak, et al., Photocheni.
Photobiol. Sci. 1:355-363 (2002). This essay was used in this example.
Plasniid cleavage was visualized by running samples on 1% agarose gels, and dainage, i.e., fonnation of relaxed, open circular fonns from closed circular foinis (undamaged) were quantified via densitometry, which permitted assessment of the protective effect of a compound.
AGE-cleavage proceeds in the absence of oxygen, and cannot be suppressed fully via antioxidants. As such, if a compound suppresses plasmid cleavage it carmot be siinply considered an aiitioxidant. In contrast, inhibition via quenching of the excited state, as described supra, nlust be presumed.
The results are presented in the Table wluch follows. Cytotoxic NaN3, which is
hereafter. Such compounds are characterized by an ability to inactivate the photoexcited state of a rinolecule which would then provoke the type of cell death described supra.
The proposed mechanism by which these molecules function is set forth in figure 9, although it is to be noted that applicants do not wish to be bound by this proposed mechanism. In brief, UV irradiation of a molecule leads to excitation of electrons (excited states, as "S*" in the figure), together with fonnation of excited singlets, and, after intersystem crossing (ISC), triplet states. These are key intermediates in the photodamage of cells. QPES compounds nullify this effect by accepting the excitation energy of the compounds associated witll photodamage by energy transfer ("ET"), neutralizing phototoxic intennediates which relax back to ground state, dissipating the energy via hannless vibrational energy, or heat. The QPES conipotulds themselves are not depleted in this process, and neutralize multiple photodainaging molecules.
It is well lulown that q>X174 plasmid is cleaved only by combined action of irradiation from solar sin-iulated liglit, and AGE-pigment enriclied protein, which acts as a UV sensitizer. AGE-BSA ("advanced glycation end product" modified bovine serum albumin), is a model for accumulation of endogenous skin sensitizers of the type described suUra. Details of the assay showing this will be found in Wondrak, et al., Photocheni.
Photobiol. Sci. 1:355-363 (2002). This essay was used in this example.
Plasniid cleavage was visualized by running samples on 1% agarose gels, and dainage, i.e., fonnation of relaxed, open circular fonns from closed circular foinis (undamaged) were quantified via densitometry, which permitted assessment of the protective effect of a compound.
AGE-cleavage proceeds in the absence of oxygen, and cannot be suppressed fully via antioxidants. As such, if a compound suppresses plasmid cleavage it carmot be siinply considered an aiitioxidant. In contrast, inhibition via quenching of the excited state, as described supra, nlust be presumed.
The results are presented in the Table wluch follows. Cytotoxic NaN3, which is
-6-laiown as a quencher of photoexcited states, was effective, as were thiol conlpounds including glutathione ("GSH"), D-penicillainine, and N-acetyl-L-cysteine ("NAC").
This assay proves the principle discussed in exalnple 7.
Table 1: Suppression of AGE-Sensitized DNA cleavage Compound % Inhibition ( SD) Catalase [400 u/mL] 57.5 4.7 SOD [300 u/hnL] 48.0 1.1 Mannitol [20 mM] 47.5 2.5 `
L-Histidine [20 mM] 43.7 0.4 NaN3 [20 mM] 94.0 0.5 DABCO [20n-iM] 32.9 1.1 DFO [1 mM] 51.3 1.0 D-Penicillamine [20 mM] 95.7+_0.7 NAC [20 inM] 99.4 0.9 GSH [20mM] 100.0 0.0 Using this assay, a series of proline related compounds and proliiie were tested. The structures of the coinpounds are set forth at figure 10. Figure 11, sets forth the results of these experiments. The test conzpoutid was added, at 100mM, and the plasmids were contacted witli one, both, or neither of UVA light (2.3 J/eni), a.nd AGE-BSA
(10 n7g/inl).
L-Pro-OCH3 was the most potent of the quenchers, but all were effective.
EXAllZPLE 8 As was discussed, suUra, photoexcited oxygen, i.e., singlet oxygen, or "102", which is a spin paired homologue of ground state, triplet oxygen or "3OZ," is the most important excited state molecule lcnown to be in involved in phototoxicity. The involvement of singlet oxygen in photooxidative daznage to, inter alia, cellular DNA, membrane lipids, and structural proteins Iike keratin and collagen, has been well docunaented. An assay was developed to determine the efficacy of coinpounds, such as proline and the proline derivatives described supra, in quenching singlet oxygen. See Lion, et al., Nature 263:442-443 (1976)õ
This assay proves the principle discussed in exalnple 7.
Table 1: Suppression of AGE-Sensitized DNA cleavage Compound % Inhibition ( SD) Catalase [400 u/mL] 57.5 4.7 SOD [300 u/hnL] 48.0 1.1 Mannitol [20 mM] 47.5 2.5 `
L-Histidine [20 mM] 43.7 0.4 NaN3 [20 mM] 94.0 0.5 DABCO [20n-iM] 32.9 1.1 DFO [1 mM] 51.3 1.0 D-Penicillamine [20 mM] 95.7+_0.7 NAC [20 inM] 99.4 0.9 GSH [20mM] 100.0 0.0 Using this assay, a series of proline related compounds and proliiie were tested. The structures of the coinpounds are set forth at figure 10. Figure 11, sets forth the results of these experiments. The test conzpoutid was added, at 100mM, and the plasmids were contacted witli one, both, or neither of UVA light (2.3 J/eni), a.nd AGE-BSA
(10 n7g/inl).
L-Pro-OCH3 was the most potent of the quenchers, but all were effective.
EXAllZPLE 8 As was discussed, suUra, photoexcited oxygen, i.e., singlet oxygen, or "102", which is a spin paired homologue of ground state, triplet oxygen or "3OZ," is the most important excited state molecule lcnown to be in involved in phototoxicity. The involvement of singlet oxygen in photooxidative daznage to, inter alia, cellular DNA, membrane lipids, and structural proteins Iike keratin and collagen, has been well docunaented. An assay was developed to determine the efficacy of coinpounds, such as proline and the proline derivatives described supra, in quenching singlet oxygen. See Lion, et al., Nature 263:442-443 (1976)õ
-7-It is well known that toluidine blue ("TB"), upon irradiation, generates `02.
In a lrnown methodology, the 102 is trapped via 2, 2, 6, 6, tetramethylpiperidine, or "TEMP." A
stable free radical, i.e., 2, 2, 6, 6 tetramethylpiperdine-l-oxyl, or "TEMPO"
forms, which can then be measured as a deten.nination of 102 generation. Key to this assay is the fact that other reactive oxygen species do not react witll TEMP to produce TEMPO, thus ensuring that the assay is specific for formation of 102.
TEMP, TB, and one of the molecules described supra were combined, and irradiated with visible light for 5 minutes, receiving a total dose of 36 J/cm2. To elaborate, 100 M TB, 7 inM TEMP, and 20 mM test compound were combined, in phosphate buffered saline, in a quarz capillary tube of 100 l volume (1.5x90 mm), TEMPO free radical signal was measured via electron paramagnetic resonance using a commercially available apparatus.
Controls were also run, to determine signal in the absence of test compound but in the presence of light, and the standard TEMPO resonance signal.
The results, presented in figure 12, panels A-G, provide data as follows.
Panel A
shows the complete singlet oxygen generating system with production of the TEMPO signal.
Panel B shows the spin signal of a commercial TEMPO reference. Panel C
demonstrates that the effects obseived in panels D-G were not caused by direct reaction of test compounds with TEMPO. Panels D & G demonstrate singlet oxygen quenching by proline derivatives and prove that derivatization of the carboxy group of proline or 4-hydroxylation does not impact quenching activity. Furtller, it shows that all of the compounds were effective quenchers.
In order to show that the quenching molecules were pllysical quenchers and were not consumed in the reaction, amino acid content of a mixture of TB and L-Pro in PBS was measured before and after a prolonged exposure to visible light (36 J/cm2), using standard reversed phase HPLC methods for amino acid analysis. No changes were observed in the peak, retention time, or AUC values, indicating chemical inertness toward the 102, providing evidence of physical quenching, rather than reactant consumption.
These experiments were designed to determine if the compounds proven to be quenchers were efficacious in protecting slcin cells.
Cultured skin fibroblasts (CF3 cells) were exposed to photooxidative stress from 1 Oz
In a lrnown methodology, the 102 is trapped via 2, 2, 6, 6, tetramethylpiperidine, or "TEMP." A
stable free radical, i.e., 2, 2, 6, 6 tetramethylpiperdine-l-oxyl, or "TEMPO"
forms, which can then be measured as a deten.nination of 102 generation. Key to this assay is the fact that other reactive oxygen species do not react witll TEMP to produce TEMPO, thus ensuring that the assay is specific for formation of 102.
TEMP, TB, and one of the molecules described supra were combined, and irradiated with visible light for 5 minutes, receiving a total dose of 36 J/cm2. To elaborate, 100 M TB, 7 inM TEMP, and 20 mM test compound were combined, in phosphate buffered saline, in a quarz capillary tube of 100 l volume (1.5x90 mm), TEMPO free radical signal was measured via electron paramagnetic resonance using a commercially available apparatus.
Controls were also run, to determine signal in the absence of test compound but in the presence of light, and the standard TEMPO resonance signal.
The results, presented in figure 12, panels A-G, provide data as follows.
Panel A
shows the complete singlet oxygen generating system with production of the TEMPO signal.
Panel B shows the spin signal of a commercial TEMPO reference. Panel C
demonstrates that the effects obseived in panels D-G were not caused by direct reaction of test compounds with TEMPO. Panels D & G demonstrate singlet oxygen quenching by proline derivatives and prove that derivatization of the carboxy group of proline or 4-hydroxylation does not impact quenching activity. Furtller, it shows that all of the compounds were effective quenchers.
In order to show that the quenching molecules were pllysical quenchers and were not consumed in the reaction, amino acid content of a mixture of TB and L-Pro in PBS was measured before and after a prolonged exposure to visible light (36 J/cm2), using standard reversed phase HPLC methods for amino acid analysis. No changes were observed in the peak, retention time, or AUC values, indicating chemical inertness toward the 102, providing evidence of physical quenching, rather than reactant consumption.
These experiments were designed to determine if the compounds proven to be quenchers were efficacious in protecting slcin cells.
Cultured skin fibroblasts (CF3 cells) were exposed to photooxidative stress from 1 Oz
-8-
9 PCT/US03/11842 which had fornled, in situ, by dye sensitization.
In brief, 50,000 fibroblasts were seeded in a 35 mm culture dish, and then treated one day later with visible light (90 seconds exposure, providing 10.8 J/cm) in the presence or absence of TB (3.3 M in Hanlcs buffered salt solution). Five minutes after treatment, cells were washed, with phosphate buffered saline. Test compound (10 mM) was present or absent in the culture during the radiation. After 3 days of cultivation, cells were harvested via trypsinization, and counted using a Coulter counter.
The results indicated that cell proliferation was highly suppressed (70%) by the combination of TB and light, but either of these alone did not do so. The compounds L-Pro-OCH3 and 4-OH-L-Pro-OCH3 showed a very clear protective effect, as is shown in figure 13 and quantified in the table which follows:
Cell Protection against Sensitized Photodamage [% SD, n=31 QPES - compound: L-Pro 6.2 5.4 L-Pro-OCH3 83.8 10.9 4-OH-L-Pro 7.3 2.5 4-OH-L-Pro-OCH3 71.0 13.4 Protection was determined via the formula:
protection 1%proliferation ((TB+Light+QPES) - % proliferation(TB+ Light) x ( (~ O
% proliferation(Control) - % proliferation(TB+Light) The more active compounds in these experiments were the ester compounds. As is showni in figure 10, when the log P values of these compounds were determined, using methods well known in the art, the esters had significantly higher log P
values than the non-esterified compounds. As the higher log P values are indicative of greater lipophilicity, it may be the case that their superior efficacy is due to maintaining longer residence time in skin and interaction with cell membranes. Proline ester derivatives with a log P value of from about -1.00 to about +8.00 are expected to be especially useful. -9-The compound 4-OH-L-proline methyl ester, described supra, is believed to be a new compound, representative of a new group of compounds. The synthesis, of the compound and guidelines for synthesis of otlier members of this family are now set forth.
4-hydroxy-L-proline was reacted witli di-tert-butyl dicarbonate, in 1N NaOH at for 30 minutes, followed by stirring, at room temperature, for an additional 3.5 hours.
The resulting protected 4-hydroxy proline was then reacted with dimethylformamide with potassiuin carbonate and methyl iodide, at 0 C, for 30 minutes, followed by stirring for an hour, at room teniperature, to produce the methyl ester.
Deprotection of tert-butyl dicarbonate was then carried out using trifluoroacetic acid, and standard methods.
The key component in this reaction is methyl iodide. By varying the allcyl halide, esters containing alkyl groups with from 1-30, preferably 1-26 carbon atoms, can be obtained.
The foregoing examples set forth various aspects of the invention, which relate to metliods for modulating photodamage, by administering at least one substance which modulates the effect of ultraviolet light on a subject. "Modulates" as used herein, refers generically to the ability to either increase the rate of photodainage, which is useful in situations where cellular proliferation needs to be arrested or to decrease the rate of damage.
Examples of the fonner category include, e.g., psoriasis, acne, premalignant and inalignant hyperproliferative disorders such as actinic keratosis, melanoma, non-melanoma skin cancer, breast cancer, and other cancers, as well as other conditions which will be known to the skilled artisan as involving the need to reduce cellular proliferation.
It is preferred that increase in photodamage, leading to a decrease in cellular proliferation, be accomplished via the use of at least one compound which has, as its "pharmacaphore" structure, a 3-hydroxypyridine ring, and is derived from a skin component, such as collagen. Exemplary of such compounds are 3-hydroxypyridine itself, vitamin B6, and most preferably, N-alkyl3-hydroxypyridinium derivatives, such as salts, wherein the alkyl chain of the derivative coinprises at least 2 and as many as 20 carboii atoms, in a preferably straight (but optimally branched) chain, which may or may not be substituted.
In brief, 50,000 fibroblasts were seeded in a 35 mm culture dish, and then treated one day later with visible light (90 seconds exposure, providing 10.8 J/cm) in the presence or absence of TB (3.3 M in Hanlcs buffered salt solution). Five minutes after treatment, cells were washed, with phosphate buffered saline. Test compound (10 mM) was present or absent in the culture during the radiation. After 3 days of cultivation, cells were harvested via trypsinization, and counted using a Coulter counter.
The results indicated that cell proliferation was highly suppressed (70%) by the combination of TB and light, but either of these alone did not do so. The compounds L-Pro-OCH3 and 4-OH-L-Pro-OCH3 showed a very clear protective effect, as is shown in figure 13 and quantified in the table which follows:
Cell Protection against Sensitized Photodamage [% SD, n=31 QPES - compound: L-Pro 6.2 5.4 L-Pro-OCH3 83.8 10.9 4-OH-L-Pro 7.3 2.5 4-OH-L-Pro-OCH3 71.0 13.4 Protection was determined via the formula:
protection 1%proliferation ((TB+Light+QPES) - % proliferation(TB+ Light) x ( (~ O
% proliferation(Control) - % proliferation(TB+Light) The more active compounds in these experiments were the ester compounds. As is showni in figure 10, when the log P values of these compounds were determined, using methods well known in the art, the esters had significantly higher log P
values than the non-esterified compounds. As the higher log P values are indicative of greater lipophilicity, it may be the case that their superior efficacy is due to maintaining longer residence time in skin and interaction with cell membranes. Proline ester derivatives with a log P value of from about -1.00 to about +8.00 are expected to be especially useful. -9-The compound 4-OH-L-proline methyl ester, described supra, is believed to be a new compound, representative of a new group of compounds. The synthesis, of the compound and guidelines for synthesis of otlier members of this family are now set forth.
4-hydroxy-L-proline was reacted witli di-tert-butyl dicarbonate, in 1N NaOH at for 30 minutes, followed by stirring, at room temperature, for an additional 3.5 hours.
The resulting protected 4-hydroxy proline was then reacted with dimethylformamide with potassiuin carbonate and methyl iodide, at 0 C, for 30 minutes, followed by stirring for an hour, at room teniperature, to produce the methyl ester.
Deprotection of tert-butyl dicarbonate was then carried out using trifluoroacetic acid, and standard methods.
The key component in this reaction is methyl iodide. By varying the allcyl halide, esters containing alkyl groups with from 1-30, preferably 1-26 carbon atoms, can be obtained.
The foregoing examples set forth various aspects of the invention, which relate to metliods for modulating photodamage, by administering at least one substance which modulates the effect of ultraviolet light on a subject. "Modulates" as used herein, refers generically to the ability to either increase the rate of photodainage, which is useful in situations where cellular proliferation needs to be arrested or to decrease the rate of damage.
Examples of the fonner category include, e.g., psoriasis, acne, premalignant and inalignant hyperproliferative disorders such as actinic keratosis, melanoma, non-melanoma skin cancer, breast cancer, and other cancers, as well as other conditions which will be known to the skilled artisan as involving the need to reduce cellular proliferation.
It is preferred that increase in photodamage, leading to a decrease in cellular proliferation, be accomplished via the use of at least one compound which has, as its "pharmacaphore" structure, a 3-hydroxypyridine ring, and is derived from a skin component, such as collagen. Exemplary of such compounds are 3-hydroxypyridine itself, vitamin B6, and most preferably, N-alkyl3-hydroxypyridinium derivatives, such as salts, wherein the alkyl chain of the derivative coinprises at least 2 and as many as 20 carboii atoms, in a preferably straight (but optimally branched) chain, which may or may not be substituted.
-10-More preferably, the allcyl group contains 2-10 carbon atoms m a straight chain, and most preferably, 2-5 carbon atoms. As was shown via the data set forth, supra, the N-ethyl derivative is especially preferred. Longer N-allcyl derivatives, which enhance the length of time the compound resides in the skin may be preferred in particular situations, sucli as, but not being limited to, situations wliere topical delivery of the compound is desired.
These molecules may be combined with, e.g., standard pharmaceutical ingredients and carriers, such as those employed in creains, lotions, shampoos, sprays, patches, or any of the known pharmacological delivery systems that are used for administering therapeutic agents to skin.
Where targeted therapy is especially desirable, the active ingredients may be attached to a second molecule which may be larger, to improve targeted delivery. Such larger molecules include, e.g., antibodies, ligands for receptors, hormones, and other molecules which either target and/or are taken up selectively by cells.
A further feature of the invention is the quenching feature of photodamage. As was shown, supra, proline and derivatives thereof are effective quenchers of photodamage.
Especially efficacious are allcyl esters of proline, such as L-Pro-OCH3 and 4-OH-L-proline;
however, other compounds are useful, as has been shown. The modes of application are precisely as is described for the enhancers discussed, supra.
Other features of the invention will be clear to the skilled artisan, and need not be reiterated herein.
These molecules may be combined with, e.g., standard pharmaceutical ingredients and carriers, such as those employed in creains, lotions, shampoos, sprays, patches, or any of the known pharmacological delivery systems that are used for administering therapeutic agents to skin.
Where targeted therapy is especially desirable, the active ingredients may be attached to a second molecule which may be larger, to improve targeted delivery. Such larger molecules include, e.g., antibodies, ligands for receptors, hormones, and other molecules which either target and/or are taken up selectively by cells.
A further feature of the invention is the quenching feature of photodamage. As was shown, supra, proline and derivatives thereof are effective quenchers of photodamage.
Especially efficacious are allcyl esters of proline, such as L-Pro-OCH3 and 4-OH-L-proline;
however, other compounds are useful, as has been shown. The modes of application are precisely as is described for the enhancers discussed, supra.
Other features of the invention will be clear to the skilled artisan, and need not be reiterated herein.
-11-
Claims (5)
1. Use of a photodamage modulator for treating photodamage caused by ultraviolet light in a human subject in need thereof, wherein said modulator is proline, 4-hydroxyproline or an alkyl ester thereof.
2. The use of claim 1, wherein said modulator inhibits photodamage.
3. The use of claim 1, wherein said proline alkyl ester comprises an alkyl moiety of 1-24 carbon atoms.
4. The use of claim 3, wherein said proline alkyl ester is L-proline-OCH3 or 4-OH-L-Pro-OCH3.
5. The use of any one of claims 1 to 4, wherein said modulator is in the form of a cream, lotion, shampoo, spray or patch.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA2668311A CA2668311C (en) | 2002-04-19 | 2003-04-17 | Enhancing photodamage caused by ultraviolet light using a 3-hydroxypyridine analog |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US37403302P | 2002-04-19 | 2002-04-19 | |
US60/374,033 | 2002-04-19 | ||
PCT/US2003/011842 WO2003088909A2 (en) | 2002-04-19 | 2003-04-17 | Methods for modulating phototoxicity |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2668311A Division CA2668311C (en) | 2002-04-19 | 2003-04-17 | Enhancing photodamage caused by ultraviolet light using a 3-hydroxypyridine analog |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2482320A1 CA2482320A1 (en) | 2003-10-30 |
CA2482320C true CA2482320C (en) | 2010-02-09 |
Family
ID=29251125
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2482320A Expired - Fee Related CA2482320C (en) | 2002-04-19 | 2003-04-17 | Methods for modulating phototoxicity |
CA2668311A Expired - Fee Related CA2668311C (en) | 2002-04-19 | 2003-04-17 | Enhancing photodamage caused by ultraviolet light using a 3-hydroxypyridine analog |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2668311A Expired - Fee Related CA2668311C (en) | 2002-04-19 | 2003-04-17 | Enhancing photodamage caused by ultraviolet light using a 3-hydroxypyridine analog |
Country Status (17)
Country | Link |
---|---|
US (3) | US6992071B2 (en) |
EP (1) | EP1496891B1 (en) |
JP (2) | JP4527405B2 (en) |
CN (1) | CN1298321C (en) |
AT (1) | ATE516802T1 (en) |
AU (1) | AU2003234119B2 (en) |
CA (2) | CA2482320C (en) |
CY (1) | CY1112194T1 (en) |
DK (1) | DK1496891T3 (en) |
ES (1) | ES2367258T3 (en) |
HK (1) | HK1077761B (en) |
IL (2) | IL164560A0 (en) |
MX (1) | MXPA04010228A (en) |
NZ (1) | NZ535982A (en) |
PT (1) | PT1496891E (en) |
RU (1) | RU2301800C2 (en) |
WO (1) | WO2003088909A2 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1298321C (en) * | 2002-04-19 | 2007-02-07 | 亚利桑那大学董事会 | Methods for modulating phototoxicity |
WO2009112360A1 (en) * | 2008-03-10 | 2009-09-17 | Basf Se | Encapsulated phase change materials in seed coatings |
JP5727364B2 (en) * | 2009-03-30 | 2015-06-03 | 株式会社 資生堂 | UV damage reducing composition |
Family Cites Families (27)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4310461A (en) * | 1980-06-23 | 1982-01-12 | E. R. Squibb & Sons, Inc. | Imido, amido and amino derivatives of mercaptoacyl prolines and pipecolic acids |
US4902684A (en) * | 1988-06-20 | 1990-02-20 | E. R. Squibb & Sons, Inc. | Benzazepine and benzothiazepine derivatives |
US6309440B1 (en) * | 1998-08-25 | 2001-10-30 | Thomas T. Yamashita | Method and composition for promoting and controlling growth of plants |
US5208249A (en) * | 1992-08-20 | 1993-05-04 | Clintec Nutrition Co. | Method for stimulating intracellular synthesis of glutathione using esters of L-2-oxothiazolidine-4-carboxylate |
JPH0797322A (en) * | 1993-09-29 | 1995-04-11 | Shiseido Co Ltd | Singlet oxygen elimination agent |
US5681554A (en) * | 1995-06-28 | 1997-10-28 | Cosmair, Inc. | Composition for treating hair and method for using the same |
US6013250A (en) * | 1995-06-28 | 2000-01-11 | L'oreal S. A. | Composition for treating hair against chemical and photo damage |
JP2961074B2 (en) * | 1995-09-06 | 1999-10-12 | 明治製菓株式会社 | Neovascular occlusive agents for photochemotherapy |
JPH09249567A (en) * | 1996-01-12 | 1997-09-22 | Otsuka Pharmaceut Co Ltd | Medicinal composition |
WO1997041826A1 (en) * | 1996-05-07 | 1997-11-13 | Thomas Jefferson University | Use of tempol in the prevention of photoaging |
US20010056085A1 (en) * | 1996-07-09 | 2001-12-27 | Avner Ramu | Methods of using vesicant drug formulations |
CA2266412A1 (en) * | 1996-09-18 | 1998-03-26 | William J. Sarill | Compositions containing cobalamin and amino acids |
EP0987266A4 (en) * | 1997-03-31 | 2000-07-26 | Wakamoto Pharma Co Ltd | Biphenyl derivatives and medicinal compositions |
US6103487A (en) * | 1997-08-27 | 2000-08-15 | Merck & Co., Inc. | Method of treating cancer |
FR2775595B1 (en) * | 1998-03-09 | 2000-05-05 | Seppic Sa | SYNERGISTIC COMPOSITION COMPRISING A LIPOAMINOACID STRUCTURE COMPOUND AND MENUPHAR EXTRACT |
JPH11255715A (en) * | 1998-03-13 | 1999-09-21 | Kao Corp | N,n-di-substituted aniline derivative |
US20020006418A1 (en) * | 1998-10-13 | 2002-01-17 | John Kung | Composition to enhance permeation of topical skin agents |
FR2792650B1 (en) * | 1999-04-20 | 2003-02-28 | Oreal | EQUIVALENT OF ELDERLY SKIN, ITS PREPARATION METHOD AND ITS USE |
WO2000066599A1 (en) * | 1999-04-30 | 2000-11-09 | Oberthuer Walter | Antioxidative vitamin b6 analogs |
US6333344B1 (en) * | 1999-05-05 | 2001-12-25 | Merck & Co. | Prolines as antimicrobial agents |
US20020006394A1 (en) * | 2000-02-11 | 2002-01-17 | Redmond Robert W. | Photochemical tissue bonding |
US6464992B2 (en) * | 2000-04-14 | 2002-10-15 | University Of Kentucky Research Foundation | Topical micronutrient delivery system and uses thereof |
EP1304323A4 (en) * | 2000-07-19 | 2005-01-12 | Kyowa Hakko Kogyo Kk | Preventives or remedies for atopic dermatitis |
JP2002128651A (en) * | 2000-10-25 | 2002-05-09 | Kose Corp | Photoaging inhibitor and skin care preparation characterized by comprising the same |
IL157042A0 (en) * | 2001-02-05 | 2004-02-08 | Buchanan Leonard Stephen | Multi-tapered dental files |
CN1298321C (en) * | 2002-04-19 | 2007-02-07 | 亚利桑那大学董事会 | Methods for modulating phototoxicity |
DE102005000868A1 (en) * | 2004-10-15 | 2006-04-20 | Henkel Kgaa | Compositions with inhibitors of prostaglandin and / or leukotriene synthesis in combination with stimulants of the release of cutaneous neuromediators |
-
2003
- 2003-04-17 CN CNB038087766A patent/CN1298321C/en not_active Expired - Fee Related
- 2003-04-17 DK DK03728425.4T patent/DK1496891T3/en active
- 2003-04-17 CA CA2482320A patent/CA2482320C/en not_active Expired - Fee Related
- 2003-04-17 ES ES03728425T patent/ES2367258T3/en not_active Expired - Lifetime
- 2003-04-17 JP JP2003585662A patent/JP4527405B2/en not_active Expired - Fee Related
- 2003-04-17 AT AT03728425T patent/ATE516802T1/en active
- 2003-04-17 US US10/418,629 patent/US6992071B2/en not_active Expired - Lifetime
- 2003-04-17 NZ NZ535982A patent/NZ535982A/en not_active IP Right Cessation
- 2003-04-17 PT PT03728425T patent/PT1496891E/en unknown
- 2003-04-17 WO PCT/US2003/011842 patent/WO2003088909A2/en active Application Filing
- 2003-04-17 MX MXPA04010228A patent/MXPA04010228A/en active IP Right Grant
- 2003-04-17 CA CA2668311A patent/CA2668311C/en not_active Expired - Fee Related
- 2003-04-17 AU AU2003234119A patent/AU2003234119B2/en not_active Ceased
- 2003-04-17 RU RU2004133322/04A patent/RU2301800C2/en not_active IP Right Cessation
- 2003-04-17 EP EP03728425A patent/EP1496891B1/en not_active Expired - Lifetime
-
2004
- 2004-09-09 US US10/937,537 patent/US20050042188A1/en not_active Abandoned
- 2004-10-13 IL IL16456004A patent/IL164560A0/xx unknown
-
2005
- 2005-09-13 US US11/226,615 patent/US7226937B2/en not_active Expired - Lifetime
-
2006
- 2006-01-05 HK HK06100235.9A patent/HK1077761B/en not_active IP Right Cessation
-
2010
- 2010-02-15 IL IL203968A patent/IL203968A/en active IP Right Grant
- 2010-04-28 JP JP2010103731A patent/JP5244150B2/en not_active Expired - Lifetime
-
2011
- 2011-10-14 CY CY20111100977T patent/CY1112194T1/en unknown
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Rapozzi et al. | Nitric oxide-mediated activity in anti-cancer photodynamic therapy | |
US10149907B2 (en) | Combinations of a photosensitizer with a hydrogen sulfide donor, thioredoxin inhibitor or nitroxide for use in photodynamic therapy | |
Karu et al. | Cell attachment modulation by radiation from a pulsed light diode (λ= 820 nm) and various chemicals | |
EP4299077A2 (en) | Skin enhancing compositions and methods | |
Mathew et al. | Ascorbic acid monoglucoside as antioxidant and radioprotector | |
Venkatesh et al. | One-and two-photon responsive sulfur dioxide (SO 2) donors: a combinatorial drug delivery for improved antibiotic therapy | |
US7226937B2 (en) | Methods for modulating phototoxicity | |
Yiakouvaki et al. | Caged-iron chelators a novel approach towards protecting skin cells against UVA-induced necrotic cell death | |
Calori et al. | B6 vitamers as generators and scavengers of reactive oxygen species | |
Wondrak et al. | Photosensitization of DNA damage by glycated proteins | |
Poh-Fitzpatrick | Pathogenesis and treatment of photocutaneous manifestations of the porphyrias | |
Shevchuk et al. | Effects of the inhibitors of energy metabolism, lonidamine and levamisole, on 5‐aminolevulinic‐acid‐induced photochemotherapy | |
Schothorst et al. | Photochemical damage to skin fibroblasts caused by protoporphyrin and violet light | |
Gaullier et al. | N-conjugates of 2, 5-disubstituted pyrrole and glutathione evaluation of their potency as antioxidants against photosensitization of NCTC 2544 keratinocytes by excess endogenous protoporphyrin IX | |
Dubbelman et al. | Cell models | |
JP3154834B2 (en) | Tyrosinase inhibitor | |
Konarski et al. | Investigation Of Photosensitizing Properties Of Protoporphyrin Derivatives With Aminoacid Substituents | |
Sprik et al. | In vitro photosensitization of human head and neck squamous cancer cells by dihematoporphyrin-ether | |
Krajewska et al. | The Influence of Depleted Glutathione Levels on the Photodynamic Action of Zinc Phthalocyanine in CHO K1 Cells | |
Glebov et al. | Prooxidant-antioxidant state of the organism during oxidative stress and correction of the L-arginine-NO system | |
JPH05320025A (en) | Tyrosinase activity suppressing agent, melanogenesis suppressing agent and dermatic agent for external use | |
US20050222117A1 (en) | Chlorophyll and its derivatives for cancer photodynamic therapy | |
Vassilopoulou-Sellin et al. | Hematoporphyrin can inhibit the metabolism and growth of embryonic chicken cartilage in vitro |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
MKLA | Lapsed |
Effective date: 20220301 |
|
MKLA | Lapsed |
Effective date: 20200831 |