JP7243005B2 - Nkx3.2及びその断片を有効成分として含む網膜疾患治療用の医薬組成物 - Google Patents
Nkx3.2及びその断片を有効成分として含む網膜疾患治療用の医薬組成物 Download PDFInfo
- Publication number
- JP7243005B2 JP7243005B2 JP2021515206A JP2021515206A JP7243005B2 JP 7243005 B2 JP7243005 B2 JP 7243005B2 JP 2021515206 A JP2021515206 A JP 2021515206A JP 2021515206 A JP2021515206 A JP 2021515206A JP 7243005 B2 JP7243005 B2 JP 7243005B2
- Authority
- JP
- Japan
- Prior art keywords
- retinal
- pharmaceutical composition
- fragment
- seq
- amino acid
- 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.)
- Active
Links
- 101100348671 Mus musculus Nkx3-2 gene Proteins 0.000 title claims description 151
- 239000012634 fragment Substances 0.000 title claims description 55
- 239000008194 pharmaceutical composition Substances 0.000 title claims description 33
- 208000017442 Retinal disease Diseases 0.000 title claims description 20
- 239000004480 active ingredient Substances 0.000 title claims description 19
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 43
- 150000001413 amino acids Chemical class 0.000 claims description 37
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 34
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 32
- 229920001184 polypeptide Polymers 0.000 claims description 31
- 206010064930 age-related macular degeneration Diseases 0.000 claims description 25
- 241000700605 Viruses Species 0.000 claims description 22
- 208000002780 macular degeneration Diseases 0.000 claims description 20
- 201000010099 disease Diseases 0.000 claims description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 18
- 201000010183 Papilledema Diseases 0.000 claims description 12
- 206010038886 Retinal oedema Diseases 0.000 claims description 12
- 201000011195 retinal edema Diseases 0.000 claims description 12
- 108091033319 polynucleotide Proteins 0.000 claims description 10
- 239000002157 polynucleotide Substances 0.000 claims description 10
- 102000040430 polynucleotide Human genes 0.000 claims description 10
- 208000008069 Geographic Atrophy Diseases 0.000 claims description 8
- 208000000208 Wet Macular Degeneration Diseases 0.000 claims description 8
- 206010012689 Diabetic retinopathy Diseases 0.000 claims description 6
- 241000702421 Dependoparvovirus Species 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 241000713666 Lentivirus Species 0.000 claims description 2
- 241000700584 Simplexvirus Species 0.000 claims description 2
- 241000700618 Vaccinia virus Species 0.000 claims description 2
- 238000001361 intraarterial administration Methods 0.000 claims description 2
- 238000007918 intramuscular administration Methods 0.000 claims description 2
- 238000007912 intraperitoneal administration Methods 0.000 claims description 2
- 238000001990 intravenous administration Methods 0.000 claims description 2
- 230000000699 topical effect Effects 0.000 claims description 2
- 241000701161 unidentified adenovirus Species 0.000 claims description 2
- 241001430294 unidentified retrovirus Species 0.000 claims description 2
- 241000699670 Mus sp. Species 0.000 description 40
- 210000000844 retinal pigment epithelial cell Anatomy 0.000 description 40
- 201000007737 Retinal degeneration Diseases 0.000 description 36
- 230000036542 oxidative stress Effects 0.000 description 36
- 230000004258 retinal degeneration Effects 0.000 description 36
- 230000014509 gene expression Effects 0.000 description 34
- 210000004899 c-terminal region Anatomy 0.000 description 20
- 238000012217 deletion Methods 0.000 description 19
- 230000037430 deletion Effects 0.000 description 19
- 230000030833 cell death Effects 0.000 description 18
- 208000005590 Choroidal Neovascularization Diseases 0.000 description 15
- 206010060823 Choroidal neovascularisation Diseases 0.000 description 15
- 230000003902 lesion Effects 0.000 description 13
- 230000002207 retinal effect Effects 0.000 description 13
- 230000000694 effects Effects 0.000 description 12
- 210000003583 retinal pigment epithelium Anatomy 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 11
- 210000001525 retina Anatomy 0.000 description 11
- 230000003247 decreasing effect Effects 0.000 description 9
- 238000011532 immunohistochemical staining Methods 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 238000010172 mouse model Methods 0.000 description 9
- 230000002018 overexpression Effects 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 230000004393 visual impairment Effects 0.000 description 9
- 238000001262 western blot Methods 0.000 description 9
- 201000004569 Blindness Diseases 0.000 description 8
- 238000010586 diagram Methods 0.000 description 8
- 238000012014 optical coherence tomography Methods 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 238000012790 confirmation Methods 0.000 description 7
- 230000034994 death Effects 0.000 description 7
- 210000004897 n-terminal region Anatomy 0.000 description 7
- 230000026683 transduction Effects 0.000 description 7
- 238000010361 transduction Methods 0.000 description 7
- 238000011740 C57BL/6 mouse Methods 0.000 description 6
- 206010025421 Macule Diseases 0.000 description 6
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 6
- 210000001508 eye Anatomy 0.000 description 6
- 230000004382 visual function Effects 0.000 description 6
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 5
- 125000003412 L-alanyl group Chemical group [H]N([H])[C@@](C([H])([H])[H])(C(=O)[*])[H] 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 230000032683 aging Effects 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 210000004204 blood vessel Anatomy 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 206010003694 Atrophy Diseases 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 101710179684 Poly [ADP-ribose] polymerase Proteins 0.000 description 4
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 4
- 238000002583 angiography Methods 0.000 description 4
- 230000037444 atrophy Effects 0.000 description 4
- 210000005252 bulbus oculi Anatomy 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 101150036876 cre gene Proteins 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 208000032843 Hemorrhage Diseases 0.000 description 3
- 101000729271 Homo sapiens Retinoid isomerohydrolase Proteins 0.000 description 3
- 125000000570 L-alpha-aspartyl group Chemical group [H]OC(=O)C([H])([H])[C@]([H])(N([H])[H])C(*)=O 0.000 description 3
- 102100031176 Retinoid isomerohydrolase Human genes 0.000 description 3
- 206010047571 Visual impairment Diseases 0.000 description 3
- 230000022534 cell killing Effects 0.000 description 3
- 210000003161 choroid Anatomy 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002571 electroretinography Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000004438 eyesight Effects 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 208000029257 vision disease Diseases 0.000 description 3
- 230000004304 visual acuity Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010040476 FITC-annexin A5 Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 125000003440 L-leucyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] 0.000 description 2
- 125000002842 L-seryl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])O[H] 0.000 description 2
- 241000872931 Myoporum sandwicense Species 0.000 description 2
- 102000018120 Recombinases Human genes 0.000 description 2
- 108010091086 Recombinases Proteins 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 230000004452 decreased vision Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 210000005081 epithelial layer Anatomy 0.000 description 2
- 230000001747 exhibiting effect Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- 210000000608 photoreceptor cell Anatomy 0.000 description 2
- 239000000790 retinal pigment Substances 0.000 description 2
- 210000001210 retinal vessel Anatomy 0.000 description 2
- 239000011697 sodium iodate Substances 0.000 description 2
- 235000015281 sodium iodate Nutrition 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 125000003831 tetrazolyl group Chemical group 0.000 description 2
- 230000009261 transgenic effect Effects 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 239000013607 AAV vector Substances 0.000 description 1
- 241001655883 Adeno-associated virus - 1 Species 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010058030 Cartilage hypertrophy Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 206010013883 Dwarfism Diseases 0.000 description 1
- 102100028091 Homeobox protein Nkx-3.2 Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000578251 Homo sapiens Homeobox protein Nkx-3.2 Proteins 0.000 description 1
- 125000001176 L-lysyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C([H])([H])C([H])([H])C([H])([H])C(N([H])[H])([H])[H] 0.000 description 1
- 125000000769 L-threonyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])[C@](O[H])(C([H])([H])[H])[H] 0.000 description 1
- 125000003798 L-tyrosyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C([H])([H])C1=C([H])C([H])=C(O[H])C([H])=C1[H] 0.000 description 1
- 125000003580 L-valyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(C([H])([H])[H])(C([H])([H])[H])[H] 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 208000022873 Ocular disease Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 206010038923 Retinopathy Diseases 0.000 description 1
- 206010038934 Retinopathy proliferative Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004155 blood-retinal barrier Anatomy 0.000 description 1
- 230000004378 blood-retinal barrier Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 229920006317 cationic polymer Polymers 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000011227 chondrocyte hypertrophy Effects 0.000 description 1
- 239000002872 contrast media Substances 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 230000006126 farnesylation Effects 0.000 description 1
- 230000008717 functional decline Effects 0.000 description 1
- ZZUFCTLCJUWOSV-UHFFFAOYSA-N furosemide Chemical compound C1=C(Cl)C(S(=O)(=O)N)=CC(C(O)=O)=C1NCC1=CC=CO1 ZZUFCTLCJUWOSV-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 238000007489 histopathology method Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 208000018769 loss of vision Diseases 0.000 description 1
- 231100000864 loss of vision Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- NALMPLUMOWIVJC-UHFFFAOYSA-N n,n,4-trimethylbenzeneamine oxide Chemical compound CC1=CC=C([N+](C)(C)[O-])C=C1 NALMPLUMOWIVJC-UHFFFAOYSA-N 0.000 description 1
- 210000001328 optic nerve Anatomy 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000011867 re-evaluation Methods 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000009528 severe injury Effects 0.000 description 1
- 230000012488 skeletal system development Effects 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229940032753 sodium iodate Drugs 0.000 description 1
- 239000002047 solid lipid nanoparticle Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000019635 sulfation Effects 0.000 description 1
- 238000005670 sulfation reaction Methods 0.000 description 1
- 230000036561 sun exposure Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/35—Animals modified by environmental factors, e.g. temperature, O2
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/072—Animals genetically altered by homologous recombination maintaining or altering function, i.e. knock in
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Ophthalmology & Optometry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
N末端伸長ドメイン-コアドメイン-C末端伸長ドメイン(I)
で表されるポリペプチドである、網膜疾患の予防又は治療用の医薬組成物を提供する。
N末端伸長ドメインは、配列番号3のアミノ酸配列を有するポリペプチドであって、配列番号3の1位のアミノ酸から開始してN末端からC末端方向に1個~42個のアミノ酸が連続的に欠失することができ、
C末端伸長ドメインは、配列番号5のアミノ酸配列を有するポリペプチドであって、配列番号5の24位のアミノ酸から開始してC末端からN末端方向に1個~23個のアミノ酸が連続的に欠失することができる。
N末端伸長ドメイン-コアドメイン-C末端伸長ドメイン(I)
で表されるポリペプチドであってもよい。
N末端伸長ドメイン-コアドメイン-C末端伸長ドメイン(I)
で表されるポリペプチドであってもよい。
C57BL/6マウスを購入し、承認された動物施設で特定の無菌状態で保管した。所定の実験プロトコルによって正常C57BL/6マウスの眼球を摘出して組職を分離した。
眼球網膜色素上皮層においてNkx3.2タンパク質の発現が報告されていないので、免疫組織化学染色(immunohistochemistry staining,IHC staining)及びウエスタンブロット分析法(western blot analysis)を用いてこれに対する検証を行った。
免疫組織化学染色(immunohistochemistry staining,IHC staining)及びウエスタンブロット分析法(western blot analysis)を用いて酸化ストレスによる網膜変性モデルにおけるNkx3.2タンパク質の発現を確認した。
本研究チームは、先行論文でCre依存的Nkx3.2過剰発現マウスを発表した(Jeong,Da-Un,Je-Yong Choi,and Dae-Won Kim.“Cartilage-Specific and Cre-Dependent Nkx3.2 Overexpression In Vivo Causes Skeletal Dwarfism by Delaying Cartilage Hypertrophy.”Journal of cellular physiology 232.1(2017):78-90.)。これに基づいて網膜色素上皮細胞に特異的なCre発現を示すBEST1-Creマウスを用いて、Cre遺伝子組換え酵素によるNkx3.2過剰発現がただ網膜色素上皮細胞のみで起こるようにした。当該マウスの作製戦略を図式化して図7に示した。
前記実験例3で作製した網膜色素上皮細胞に特異的なNkx3.2過剰発現マウスでNaIO3を用いて酸化ストレスによる網膜変性を誘導した。
酸化ストレスによって網膜変性が誘導されたマウスの半球形の網膜をフラットマウントして網膜変性の病変を確認した。その結果を図9に示した。
酸化ストレスによって網膜変性が誘導されたマウスの眼球網膜構造を眼底画像撮影方法により分析した。その結果を図10に示した。
酸化ストレスによって網膜変性が誘導されたマウスの眼球網膜構造を光干渉断層撮影方法により分析した。その結果を図11に示した。
酸化ストレスによって網膜変性が誘導されたマウスの眼球網膜構造を組職学的に分析した。その結果を図12に示した。
網膜電図検査(electroretinography,ERG)によってNkx3.2過剰発現による視覚機能の保存効果を確認した。
ヒト網膜色素上皮細胞に400μM、600μM、800μM、1mMの過酸化水素(H2O2)をそれぞれ処理して酸化ストレスを誘導した。その後、WST分析手法(water-soluble tetrazolium salt assay)を用いて酸化ストレスを誘導するH2O2濃度による細胞生存率を測定した。その結果を図14に示した。
ヒト網膜色素上皮細胞に800μMのH2O2を処理して酸化ストレスを誘導し、前記酸化ストレスが誘導された細胞を2つのグループに分けた。1つのグループにはNkx3.2発現ウイルス(Lenti-Nkx3.2)を投与し、他のグループにはNkx3.2発現抑制ウイルス(sh-Nkx3.2)を投与した。その後、アネキシンV-FITC死滅キットを用いて各ウイルスの投与によるヒト網膜色素上皮細胞の酸化ストレスによる細胞死滅率を測定した。その結果を図17及び図18に示した。
実験例8.1.酸化ストレスによるNkx3.2全長及びその断片の網膜色素上皮細胞の死滅抑制能の評価
Nkx3.2全長及びその断片が酸化ストレスであるH2O2によって誘導される網膜色素上皮細胞の死滅を抑制する機能を有しているかどうかを評価するために、ARPE19細胞が播種された12ウェルプレート(3E+4cell/well)にhNkx3.2全長(配列番号7)、及び配列番号9(hNkx3.2全長配列のアミノ酸1~320)、配列番号10(hNkx3.2全長配列のアミノ酸1~307)、配列番号12(hNkx3.2全長配列のアミノ酸99~333)、配列番号13(hNkx3.2全長配列のアミノ酸112~333)、配列番号20(hNkx3.2全長配列のアミノ酸112~320)、配列番号21(hNkx3.2全長配列のアミノ酸123~320)、配列番号23(hNkx3.2全長配列のアミノ酸150~320)及び配列番号28(hNkx3.2全長配列のアミノ酸165~310)のアミノ酸配列を有するhNkx3.2の断片をコードするDNAプラスミドを各0.6ugずつ形質導入した。
形質導入の特性上、100%の効率が現れないため、前記実験例の細胞生存能の分析でNkx3.2全長及びその断片のH2O2による網膜色素上皮細胞の死滅抑制能が低評価された可能性がある。そこで、Nkx3.2全長及びその断片に対する形質導入の効率を免疫細胞化学技術を用いて評価した。まず、スライドガラスが載せられている12ウェルプレートにAREP19細胞を2E+4個ずつ播種した。hNkx3.2全長又はその断片をコードするDNAプラスミドをそれぞれ0.6ugずつ形質導入した。48時間後、各ウェルに2%のパラホルムアルデヒドを入れて常温で15分間反応させて細胞を固定した。その後、各ウェルに0.1%クエン酸ナトリウム内0.1%Triton X-100溶液を入れて常温で30分間反応させ、抗体が細胞内に侵入しやすい状態に作った。その後、PBSで2回洗浄した。
Nkx3.2全長及びその断片の形質導入の効率を考慮し、H2O2によるARPE19細胞死滅を抑制することができるか否かを再評価した。その結果を下記表1に示した。
表1に示したように、Nkx3.2全長を含めたNkx3.2断片は、H2O2によるARPE19細胞死滅を対照群に比べて少なくとも30%から最大75%程度まで抑制した。
網膜色素上皮細胞に特異的なNkx3.2過剰発現マウスにレーザーを照射して脈絡膜新生血管(choroidal neovascularization,CNV)を誘導した。9日後、マウスに造影剤を腹腔注射して網膜血管に対して眼底造影法(fundus fluorescence angiography,FAA)を施した。前記眼底造影法から確保した画像に基づいて脈絡膜新生血管の病変のCTFを測定し、野生型(wild type)マウスとNkx3.2過剰発現マウスとの間の脈絡膜新生血管の病変の大きさの差を比較した。その結果を図21a及び図21bに示した。
網膜色素上皮細胞に特異的なNkx3.2過剰発現マウスでレーザーを照射して脈絡膜新生血管を誘導した後、9日目に脈絡膜新生血管を誘導した網膜領域に対して光干渉断層撮影方法を施した。前記光干渉断層撮影から確保した画像に基づいて野生型マウスとNkx3.2過剰発現マウスとの間の脈絡膜新生血管の病変で引き起こされた網膜浮腫のパターンの差を比較した。その結果を図22a及び図22bに示した。
Claims (9)
- Nkx3.2又はその断片を有効成分として含む網膜疾患の予防又は治療用の医薬組成物であって、前記Nkx3.2又はその断片は、配列番号7又は12のアミノ酸配列を有するポリペプチドである、医薬組成物。
- 前記網膜疾患が、黄斑変性、糖尿病性網膜症、脈絡膜新生血管症及び網膜浮腫からなる群から選択されるいずれか1つである、請求項1に記載の医薬組成物。
- 前記黄斑変性が、乾性黄斑変性又は湿性黄斑変性である、請求項2に記載の医薬組成物。
- Nkx3.2又はその断片をコードするポリヌクレオチドが含まれた組換えウイルスを有効成分として含む、網膜疾患の予防又は治療用の医薬組成物であって、前記Nkx3.2又はその断片は、配列番号7又は12のアミノ酸配列を有するポリペプチドである、医薬組成物。
- 前記ウイルスが、アデノウイルス、アデノ随伴ウイルス(Adeno-associated viruses:AAV)、レトロウイルス、レンチウイルス、単純ヘルペスウイルス及びワクシニアウイルスからなる群から選択されるいずれか1つであることを特徴とする、請求項4に記載の医薬組成物。
- 前記網膜疾患が、黄斑変性、糖尿病性網膜症、脈絡膜新生血管症及び網膜浮腫からなる群から選択されるいずれか1つである、請求項4又は5に記載の医薬組成物。
- 前記黄斑変性が、乾性黄斑変性又は湿性黄斑変性である、請求項6に記載の医薬組成物。
- 網膜疾患の予防又は治療用の医薬組成物の製造における、Nkx3.2又はその断片の使用であって、前記Nkx3.2又はその断片は、配列番号7又は12のアミノ酸配列を有するポリペプチドである、使用。
- 前記医薬組成物は、静脈内、動脈内、腹腔内、筋肉内、胸骨内、経皮、鼻腔、吸入、局所、直腸、経口、眼球内及び皮内からなる群から選択されるいずれか1つの経路により個体に投与される、請求項8に記載の使用。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/KR2018/012428 WO2020080583A1 (ko) | 2018-10-19 | 2018-10-19 | Nkx3.2 및 이의 단편을 유효성분으로 포함하는 망막질환 치료용 약학적 조성물 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2022512554A JP2022512554A (ja) | 2022-02-07 |
JP7243005B2 true JP7243005B2 (ja) | 2023-03-22 |
Family
ID=70282952
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2021515206A Active JP7243005B2 (ja) | 2018-10-19 | 2018-10-19 | Nkx3.2及びその断片を有効成分として含む網膜疾患治療用の医薬組成物 |
Country Status (7)
Country | Link |
---|---|
US (2) | US11628203B2 (ja) |
EP (1) | EP3868393A4 (ja) |
JP (1) | JP7243005B2 (ja) |
CN (1) | CN112739370A (ja) |
AU (1) | AU2018445968A1 (ja) |
CA (1) | CA3112915A1 (ja) |
WO (1) | WO2020080583A1 (ja) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180236035A1 (en) | 2010-05-03 | 2018-08-23 | Scott & White Healthcare | Protein therapy for treatment of retinal diseases |
JP2019535310A (ja) | 2016-11-09 | 2019-12-12 | アイシーエム・カンパニー、リミテッドIcm Co.,Ltd. | Nkx3.2断片及び活性成分としてNkx3.2断片を含む医薬組成物 |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4797368A (en) | 1985-03-15 | 1989-01-10 | The United States Of America As Represented By The Department Of Health And Human Services | Adeno-associated virus as eukaryotic expression vector |
US5139941A (en) | 1985-10-31 | 1992-08-18 | University Of Florida Research Foundation, Inc. | AAV transduction vectors |
WO2001003743A1 (en) * | 1999-07-12 | 2001-01-18 | Walter Gehring | Manipulation of tissue or organ type using the notch pathway |
US8084258B2 (en) * | 1999-07-12 | 2011-12-27 | University Of Basel | Manipulation of tissue of organ type using the notch pathway |
EP2094719A4 (en) * | 2006-12-19 | 2010-01-06 | Genego Inc | NEW PROCEDURES FOR THE FUNCTIONAL ANALYSIS OF EXPERIMENTAL HIGH-PERFORMANCE DATA AND IDENTIFIED GENDER GROUPS THEREOF |
EP2415478A4 (en) * | 2009-03-30 | 2013-01-16 | Santen Pharmaceutical Co Ltd | PROPHYLAXIS OR THERAPY AGENTS FOR KIDNEY DISEASES AND METHODS OF PREVENTING OR TREATING NURSE DISEASES, EACH OF THEM WITH A JNK (C-JUN-N-TERMINAL KINASE) -IMBING PEPTIDE, AND USE OF THE PEPTIDE |
WO2011093647A2 (en) | 2010-01-26 | 2011-08-04 | Industry-Academic Cooperation Foundation, Yonsei University | Genes implicated in osteoarthritis and use thereof |
KR101150900B1 (ko) * | 2010-01-26 | 2012-05-30 | 연세대학교 산학협력단 | 퇴행성 관절염 치료 조성물 |
WO2012097057A2 (en) * | 2011-01-11 | 2012-07-19 | Tufts University | Methods, compositions and kits for modulating trans-differentiation of muscle satellite cells |
WO2018194423A1 (ko) * | 2017-04-21 | 2018-10-25 | 아이씨엠 주식회사 | Nkx3.2 및 이의 단편을 유효성분으로 포함하는 망막질환 치료용 약학적 조성물 |
CN108103104B (zh) * | 2017-12-21 | 2021-07-06 | 北京锦篮基因科技有限公司 | 一种预防和治疗脉络膜新生血管相关眼部疾病的基因药物 |
-
2018
- 2018-10-19 CN CN201880097849.1A patent/CN112739370A/zh active Pending
- 2018-10-19 WO PCT/KR2018/012428 patent/WO2020080583A1/ko unknown
- 2018-10-19 JP JP2021515206A patent/JP7243005B2/ja active Active
- 2018-10-19 AU AU2018445968A patent/AU2018445968A1/en active Pending
- 2018-10-19 EP EP18937009.1A patent/EP3868393A4/en active Pending
- 2018-10-19 CA CA3112915A patent/CA3112915A1/en active Pending
- 2018-10-19 US US17/273,546 patent/US11628203B2/en active Active
-
2022
- 2022-12-27 US US18/146,946 patent/US20230149506A1/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180236035A1 (en) | 2010-05-03 | 2018-08-23 | Scott & White Healthcare | Protein therapy for treatment of retinal diseases |
JP2019535310A (ja) | 2016-11-09 | 2019-12-12 | アイシーエム・カンパニー、リミテッドIcm Co.,Ltd. | Nkx3.2断片及び活性成分としてNkx3.2断片を含む医薬組成物 |
Also Published As
Publication number | Publication date |
---|---|
AU2018445968A1 (en) | 2021-04-15 |
WO2020080583A8 (ko) | 2020-11-26 |
CN112739370A (zh) | 2021-04-30 |
US11628203B2 (en) | 2023-04-18 |
WO2020080583A1 (ko) | 2020-04-23 |
EP3868393A4 (en) | 2022-06-08 |
CA3112915A1 (en) | 2020-04-23 |
JP2022512554A (ja) | 2022-02-07 |
US20210187065A1 (en) | 2021-06-24 |
US20230149506A1 (en) | 2023-05-18 |
EP3868393A1 (en) | 2021-08-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Kjellstrom et al. | Retinoschisin gene therapy and natural history in the Rs1h-KO mouse: long-term rescue from retinal degeneration | |
Ramírez et al. | Vasoinhibin gene transfer by adenoassociated virus type 2 protects against VEGF-and diabetes-induced retinal vasopermeability | |
KR20170137730A (ko) | 망막 추상체에 폴리뉴클레오타이드의 유리체 내 전달을 위한 조성물 및 방법 | |
JP2020535184A (ja) | 翻訳後修飾された完全ヒト抗VEGF Fabによる眼疾患の治療 | |
Dai et al. | AAV-mediated lysophosphatidylcholine acyltransferase 1 (Lpcat1) gene replacement therapy rescues retinal degeneration in rd11 mice | |
Kreppel et al. | Long-term transgene expression in the RPE after gene transfer with a high-capacity adenoviral vector | |
EP3052124A1 (en) | Use of neuroglobin agonist for preventing or treating mitochondrial rcci and/or rcciii deficiency disease | |
Wu et al. | Gene therapy for detached retina by adeno-associated virus vector expressing glial cell line–derived neurotrophic factor | |
Tuo et al. | AAV5-mediated sFLT01 gene therapy arrests retinal lesions in Ccl2−/−/Cx3cr1−/− mice | |
Dinculescu et al. | Pathological effects of mutant C1QTNF5 (S163R) expression in murine retinal pigment epithelium | |
KR102044530B1 (ko) | Nkx3.2 및 이의 단편을 유효성분으로 포함하는 망막질환 치료용 약학적 조성물 | |
Semkova et al. | Overexpression of FasL in retinal pigment epithelial cells reduces choroidal neovascularization | |
CN116406304A (zh) | 使用AIMP2-DX2和任选地miR-142的靶序列及其组合物治疗年龄相关的黄斑疾病的方法 | |
US10583150B2 (en) | Pharmaceutical composition containing mTOR inhibitor for treating macular degeneration | |
JP7243005B2 (ja) | Nkx3.2及びその断片を有効成分として含む網膜疾患治療用の医薬組成物 | |
US20230135501A1 (en) | Gene therapy | |
CN115243766A (zh) | 治疗常染色体显性bestrophin病及其评估方法 | |
KR20200044386A (ko) | Nkx3.2 및 이의 단편을 유효성분으로 포함하는 망막질환 치료용 약학적 조성물 | |
US20220331241A1 (en) | Treatment of x-linked juvenile retinoschisis | |
US8546347B2 (en) | Composition for treatment and improvement of diabetes comprising caveolin as active ingredient and a method for treatment of diabetes using it | |
Wei-Zhang et al. | Chimpanzee adenovirus-mediated multiple gene therapy for age-related macular degeneration | |
RU2778806C2 (ru) | Фармацевтическая композиция для профилактики или лечения аритмии сердца | |
WO2023218430A1 (en) | Methods of treating retinal degenerative diseases using aimp2-dx2 and optionally a target sequence for mir‑142 and compositions thereof | |
WO2023023256A1 (en) | Aav-mediated gene transfer for retinopathy | |
KR20140071455A (ko) | 망막 변성 병태의 치료를 위한 조성물 및 방법 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20210323 |
|
RD01 | Notification of change of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7426 Effective date: 20210323 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20210927 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220913 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20221124 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20230207 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230213 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7243005 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |