CN113825521A - 腺相关病毒及其用于内耳疗法的用途 - Google Patents
腺相关病毒及其用于内耳疗法的用途 Download PDFInfo
- Publication number
- CN113825521A CN113825521A CN201980085229.0A CN201980085229A CN113825521A CN 113825521 A CN113825521 A CN 113825521A CN 201980085229 A CN201980085229 A CN 201980085229A CN 113825521 A CN113825521 A CN 113825521A
- Authority
- CN
- China
- Prior art keywords
- hearing loss
- recombinant aav
- cells
- virion
- aav
- 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.)
- Pending
Links
- 210000003027 ear inner Anatomy 0.000 title claims abstract description 96
- 241000702421 Dependoparvovirus Species 0.000 title claims abstract description 16
- 238000002560 therapeutic procedure Methods 0.000 title description 3
- 210000004027 cell Anatomy 0.000 claims description 112
- 210000002845 virion Anatomy 0.000 claims description 96
- 210000002768 hair cell Anatomy 0.000 claims description 93
- 206010011878 Deafness Diseases 0.000 claims description 90
- 231100000888 hearing loss Toxicity 0.000 claims description 90
- 230000010370 hearing loss Effects 0.000 claims description 90
- 208000016354 hearing loss disease Diseases 0.000 claims description 90
- 238000000034 method Methods 0.000 claims description 72
- 150000007523 nucleic acids Chemical class 0.000 claims description 48
- 210000003477 cochlea Anatomy 0.000 claims description 36
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 36
- 108090000565 Capsid Proteins Proteins 0.000 claims description 29
- 102100023321 Ceruloplasmin Human genes 0.000 claims description 29
- -1 REST Proteins 0.000 claims description 29
- 150000001413 amino acids Chemical class 0.000 claims description 27
- 208000012886 Vertigo Diseases 0.000 claims description 26
- 108090000623 proteins and genes Proteins 0.000 claims description 26
- 231100000889 vertigo Toxicity 0.000 claims description 26
- 108020004707 nucleic acids Proteins 0.000 claims description 25
- 102000039446 nucleic acids Human genes 0.000 claims description 25
- 229920001184 polypeptide Polymers 0.000 claims description 24
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 24
- 210000003030 auditory receptor cell Anatomy 0.000 claims description 19
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 18
- 230000005779 cell damage Effects 0.000 claims description 17
- 208000037887 cell injury Diseases 0.000 claims description 17
- 238000003780 insertion Methods 0.000 claims description 17
- 230000037431 insertion Effects 0.000 claims description 17
- 230000008929 regeneration Effects 0.000 claims description 17
- 238000011069 regeneration method Methods 0.000 claims description 17
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 15
- 241000702423 Adeno-associated virus - 2 Species 0.000 claims description 14
- 201000010099 disease Diseases 0.000 claims description 14
- 210000002480 semicircular canal Anatomy 0.000 claims description 11
- 206010011903 Deafness traumatic Diseases 0.000 claims description 9
- 208000002946 Noise-Induced Hearing Loss Diseases 0.000 claims description 9
- 230000002452 interceptive effect Effects 0.000 claims description 9
- 102100033189 Diablo IAP-binding mitochondrial protein Human genes 0.000 claims description 8
- 102100037391 Gasdermin-E Human genes 0.000 claims description 8
- 101001026269 Homo sapiens Gasdermin-E Proteins 0.000 claims description 8
- 208000014674 injury Diseases 0.000 claims description 8
- 241001164825 Adeno-associated virus - 8 Species 0.000 claims description 7
- 102100040191 Alpha-tectorin Human genes 0.000 claims description 7
- 101000889766 Homo sapiens Alpha-tectorin Proteins 0.000 claims description 7
- 230000008733 trauma Effects 0.000 claims description 7
- 101100135858 Caenorhabditis elegans pde-2 gene Proteins 0.000 claims description 6
- 230000000692 anti-sense effect Effects 0.000 claims description 6
- 102100037156 Gap junction beta-2 protein Human genes 0.000 claims description 5
- 101000954092 Homo sapiens Gap junction beta-2 protein Proteins 0.000 claims description 5
- 101000591234 Homo sapiens Phosphatidylinositol phosphatase PTPRQ Proteins 0.000 claims description 5
- 101000591236 Homo sapiens Receptor-type tyrosine-protein phosphatase R Proteins 0.000 claims description 5
- 102100034105 Phosphatidylinositol phosphatase PTPRQ Human genes 0.000 claims description 5
- 102100030374 Actin, cytoplasmic 2 Human genes 0.000 claims description 4
- 101000798762 Anguilla anguilla Troponin C, skeletal muscle Proteins 0.000 claims description 4
- 102100035445 Carcinoembryonic antigen-related cell adhesion molecule 16 Human genes 0.000 claims description 4
- 102100040996 Cochlin Human genes 0.000 claims description 4
- 102100024133 Coiled-coil domain-containing protein 50 Human genes 0.000 claims description 4
- 102100033825 Collagen alpha-1(XI) chain Human genes 0.000 claims description 4
- 102100033885 Collagen alpha-2(XI) chain Human genes 0.000 claims description 4
- 108010069176 Connexin 30 Proteins 0.000 claims description 4
- 101150082208 DIABLO gene Proteins 0.000 claims description 4
- 102100030960 DNA replication licensing factor MCM2 Human genes 0.000 claims description 4
- 102100030910 Eyes absent homolog 4 Human genes 0.000 claims description 4
- 102100039397 Gap junction beta-3 protein Human genes 0.000 claims description 4
- 102100039401 Gap junction beta-6 protein Human genes 0.000 claims description 4
- 102100034227 Grainyhead-like protein 2 homolog Human genes 0.000 claims description 4
- 102100029279 Homeobox protein SIX1 Human genes 0.000 claims description 4
- 102100023605 Homer protein homolog 2 Human genes 0.000 claims description 4
- 101000773237 Homo sapiens Actin, cytoplasmic 2 Proteins 0.000 claims description 4
- 101000737645 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 16 Proteins 0.000 claims description 4
- 101000748988 Homo sapiens Cochlin Proteins 0.000 claims description 4
- 101000910772 Homo sapiens Coiled-coil domain-containing protein 50 Proteins 0.000 claims description 4
- 101000710623 Homo sapiens Collagen alpha-1(XI) chain Proteins 0.000 claims description 4
- 101000710619 Homo sapiens Collagen alpha-2(XI) chain Proteins 0.000 claims description 4
- 101000583807 Homo sapiens DNA replication licensing factor MCM2 Proteins 0.000 claims description 4
- 101001018431 Homo sapiens DNA replication licensing factor MCM7 Proteins 0.000 claims description 4
- 101000871228 Homo sapiens Diablo IAP-binding mitochondrial protein Proteins 0.000 claims description 4
- 101000938422 Homo sapiens Eyes absent homolog 4 Proteins 0.000 claims description 4
- 101000889136 Homo sapiens Gap junction beta-3 protein Proteins 0.000 claims description 4
- 101001069929 Homo sapiens Grainyhead-like protein 2 homolog Proteins 0.000 claims description 4
- 101000634171 Homo sapiens Homeobox protein SIX1 Proteins 0.000 claims description 4
- 101001048464 Homo sapiens Homer protein homolog 2 Proteins 0.000 claims description 4
- 101000599613 Homo sapiens Interferon lambda receptor 1 Proteins 0.000 claims description 4
- 101000745406 Homo sapiens Ketimine reductase mu-crystallin Proteins 0.000 claims description 4
- 101001038339 Homo sapiens LIM homeobox transcription factor 1-alpha Proteins 0.000 claims description 4
- 101000588972 Homo sapiens Myosin-1 Proteins 0.000 claims description 4
- 101001030232 Homo sapiens Myosin-9 Proteins 0.000 claims description 4
- 101000720696 Homo sapiens Oxysterol-binding protein-related protein 2 Proteins 0.000 claims description 4
- 101000614335 Homo sapiens P2X purinoceptor 2 Proteins 0.000 claims description 4
- 101001094737 Homo sapiens POU domain, class 4, transcription factor 3 Proteins 0.000 claims description 4
- 101000994648 Homo sapiens Potassium voltage-gated channel subfamily KQT member 4 Proteins 0.000 claims description 4
- 101000928791 Homo sapiens Protein diaphanous homolog 1 Proteins 0.000 claims description 4
- 101001133085 Homo sapiens Sialomucin core protein 24 Proteins 0.000 claims description 4
- 101000666340 Homo sapiens Tenascin Proteins 0.000 claims description 4
- 101000785523 Homo sapiens Tight junction protein ZO-2 Proteins 0.000 claims description 4
- 101000801040 Homo sapiens Transmembrane channel-like protein 1 Proteins 0.000 claims description 4
- 101000803332 Homo sapiens Wolframin Proteins 0.000 claims description 4
- 102100037971 Interferon lambda receptor 1 Human genes 0.000 claims description 4
- 102100039386 Ketimine reductase mu-crystallin Human genes 0.000 claims description 4
- 102100040290 LIM homeobox transcription factor 1-alpha Human genes 0.000 claims description 4
- 108010009047 Myosin VIIa Proteins 0.000 claims description 4
- 102100032975 Myosin-1 Human genes 0.000 claims description 4
- 102100038938 Myosin-9 Human genes 0.000 claims description 4
- 102100022691 NACHT, LRR and PYD domains-containing protein 3 Human genes 0.000 claims description 4
- 102100025925 Oxysterol-binding protein-related protein 2 Human genes 0.000 claims description 4
- 102100040479 P2X purinoceptor 2 Human genes 0.000 claims description 4
- 102100035398 POU domain, class 4, transcription factor 3 Human genes 0.000 claims description 4
- 102100034363 Potassium voltage-gated channel subfamily KQT member 4 Human genes 0.000 claims description 4
- 102100036490 Protein diaphanous homolog 1 Human genes 0.000 claims description 4
- 102100025799 Protein phosphatase 1K, mitochondrial Human genes 0.000 claims description 4
- 108010001946 Pyrin Domain-Containing 3 Protein NLR Family Proteins 0.000 claims description 4
- 108091006282 SLC17A8 Proteins 0.000 claims description 4
- 102100034258 Sialomucin core protein 24 Human genes 0.000 claims description 4
- 108020004459 Small interfering RNA Proteins 0.000 claims description 4
- 102100038126 Tenascin Human genes 0.000 claims description 4
- 102100026637 Tight junction protein ZO-2 Human genes 0.000 claims description 4
- 102100033690 Transmembrane channel-like protein 1 Human genes 0.000 claims description 4
- 102100031834 Unconventional myosin-VI Human genes 0.000 claims description 4
- 102100031835 Unconventional myosin-VIIa Human genes 0.000 claims description 4
- 102100038033 Vesicular glutamate transporter 3 Human genes 0.000 claims description 4
- 102100036022 Wolframin Human genes 0.000 claims description 4
- 108010049787 myosin VI Proteins 0.000 claims description 4
- 239000004055 small Interfering RNA Substances 0.000 claims description 4
- HEZMWWAKWCSUCB-PHDIDXHHSA-N (3R,4R)-3,4-dihydroxycyclohexa-1,5-diene-1-carboxylic acid Chemical compound O[C@@H]1C=CC(C(O)=O)=C[C@H]1O HEZMWWAKWCSUCB-PHDIDXHHSA-N 0.000 claims description 3
- ITHCSGCUQDMYAI-ZMIZWQJLSA-N 2-carboxy-D-arabinitol 1,5-bisphosphate Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@](O)(COP(O)(O)=O)C(O)=O ITHCSGCUQDMYAI-ZMIZWQJLSA-N 0.000 claims description 3
- 241001634120 Adeno-associated virus - 5 Species 0.000 claims description 3
- 102100040999 Catechol O-methyltransferase Human genes 0.000 claims description 3
- 108020002739 Catechol O-methyltransferase Proteins 0.000 claims description 3
- 102100032363 Choline dehydrogenase, mitochondrial Human genes 0.000 claims description 3
- 101710181272 Choline dehydrogenase, mitochondrial Proteins 0.000 claims description 3
- 102100035347 DmX-like protein 2 Human genes 0.000 claims description 3
- 102100025734 Dual specificity protein phosphatase CDC14A Human genes 0.000 claims description 3
- 102100031809 Espin Human genes 0.000 claims description 3
- 101000804534 Homo sapiens DmX-like protein 2 Proteins 0.000 claims description 3
- 101000932600 Homo sapiens Dual specificity protein phosphatase CDC14A Proteins 0.000 claims description 3
- 101000920837 Homo sapiens Espin Proteins 0.000 claims description 3
- 101000898034 Homo sapiens Hepatocyte growth factor Proteins 0.000 claims description 3
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 claims description 3
- 101001116774 Homo sapiens Methionine-R-sulfoxide reductase B2, mitochondrial Proteins 0.000 claims description 3
- 101001134172 Homo sapiens Otoancorin Proteins 0.000 claims description 3
- 101001134169 Homo sapiens Otoferlin Proteins 0.000 claims description 3
- 101001134207 Homo sapiens Otogelin Proteins 0.000 claims description 3
- 101001134210 Homo sapiens Otogelin-like protein Proteins 0.000 claims description 3
- 101000886818 Homo sapiens PDZ domain-containing protein GIPC1 Proteins 0.000 claims description 3
- 101000868152 Homo sapiens Son of sevenless homolog 1 Proteins 0.000 claims description 3
- 101000585180 Homo sapiens Stereocilin Proteins 0.000 claims description 3
- 101000851696 Homo sapiens Steroid hormone receptor ERR2 Proteins 0.000 claims description 3
- 101000713234 Homo sapiens TRIO and F-actin-binding protein Proteins 0.000 claims description 3
- 101000764625 Homo sapiens Transmembrane inner ear expressed protein Proteins 0.000 claims description 3
- 101000585635 Homo sapiens Unconventional myosin-XV Proteins 0.000 claims description 3
- 101000666127 Homo sapiens Whirlin Proteins 0.000 claims description 3
- 101710173431 L-carnitine dehydrogenase Proteins 0.000 claims description 3
- 101150116611 LRRC51 gene Proteins 0.000 claims description 3
- 102100022186 Leucine-rich repeat-containing protein 51 Human genes 0.000 claims description 3
- 101150117157 MYO3 gene Proteins 0.000 claims description 3
- 102100024862 Methionine-R-sulfoxide reductase B2, mitochondrial Human genes 0.000 claims description 3
- 101100235513 Mus musculus Lhfpl1 gene Proteins 0.000 claims description 3
- 101100079084 Mus musculus Myo7a gene Proteins 0.000 claims description 3
- 101100403745 Mus musculus Myot gene Proteins 0.000 claims description 3
- 102100034199 Otoancorin Human genes 0.000 claims description 3
- 102100034198 Otoferlin Human genes 0.000 claims description 3
- 102100034205 Otogelin Human genes 0.000 claims description 3
- 102100034206 Otogelin-like protein Human genes 0.000 claims description 3
- 101710105116 Oxygen-dependent choline dehydrogenase Proteins 0.000 claims description 3
- 102100039983 PDZ domain-containing protein GIPC1 Human genes 0.000 claims description 3
- 102100032421 Protein S100-A6 Human genes 0.000 claims description 3
- 101710122252 Protein S100-G Proteins 0.000 claims description 3
- 102100029924 Stereocilin Human genes 0.000 claims description 3
- 102100036831 Steroid hormone receptor ERR2 Human genes 0.000 claims description 3
- 102100036855 TRIO and F-actin-binding protein Human genes 0.000 claims description 3
- 101150104365 Tomt gene Proteins 0.000 claims description 3
- 102100026225 Transmembrane inner ear expressed protein Human genes 0.000 claims description 3
- 102100029836 Unconventional myosin-XV Human genes 0.000 claims description 3
- 201000008616 Usher syndrome type 1 Diseases 0.000 claims description 3
- 102100038102 Whirlin Human genes 0.000 claims description 3
- 108091070501 miRNA Proteins 0.000 claims description 3
- 239000002679 microRNA Substances 0.000 claims description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims 2
- 239000005090 green fluorescent protein Substances 0.000 description 76
- 208000015181 infectious disease Diseases 0.000 description 59
- 241000699670 Mus sp. Species 0.000 description 38
- 239000000047 product Substances 0.000 description 33
- 208000002267 Anti-neutrophil cytoplasmic antibody-associated vasculitis Diseases 0.000 description 31
- 239000007924 injection Substances 0.000 description 26
- 238000002347 injection Methods 0.000 description 26
- 108091028043 Nucleic acid sequence Proteins 0.000 description 25
- 239000000203 mixture Substances 0.000 description 24
- 210000000067 inner hair cell Anatomy 0.000 description 22
- 238000001415 gene therapy Methods 0.000 description 19
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 18
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 18
- 239000013607 AAV vector Substances 0.000 description 14
- 101150083557 Ear gene Proteins 0.000 description 12
- 241001465754 Metazoa Species 0.000 description 11
- 239000002245 particle Substances 0.000 description 11
- 230000001720 vestibular Effects 0.000 description 11
- 230000000694 effects Effects 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 210000004307 hair cells vestibular Anatomy 0.000 description 9
- 229920002477 rna polymer Polymers 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 239000013603 viral vector Substances 0.000 description 9
- 239000008194 pharmaceutical composition Substances 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 238000005096 rolling process Methods 0.000 description 8
- 208000024891 symptom Diseases 0.000 description 8
- 239000013598 vector Substances 0.000 description 8
- 230000003612 virological effect Effects 0.000 description 8
- 241000700605 Viruses Species 0.000 description 7
- 230000006378 damage Effects 0.000 description 7
- 210000003780 hair follicle Anatomy 0.000 description 7
- 230000009467 reduction Effects 0.000 description 7
- 230000004044 response Effects 0.000 description 7
- 208000002173 dizziness Diseases 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 102100031036 Leucine-rich repeat-containing G-protein coupled receptor 5 Human genes 0.000 description 5
- 241000124008 Mammalia Species 0.000 description 5
- 238000000540 analysis of variance Methods 0.000 description 5
- 210000000234 capsid Anatomy 0.000 description 5
- 238000001476 gene delivery Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 238000011002 quantification Methods 0.000 description 5
- 230000000638 stimulation Effects 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- 102000053602 DNA Human genes 0.000 description 4
- 101001063456 Homo sapiens Leucine-rich repeat-containing G-protein coupled receptor 5 Proteins 0.000 description 4
- 230000002411 adverse Effects 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 210000000133 brain stem Anatomy 0.000 description 4
- 239000002771 cell marker Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000001186 cumulative effect Effects 0.000 description 4
- 208000031513 cyst Diseases 0.000 description 4
- 208000035475 disorder Diseases 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 108020004705 Codon Proteins 0.000 description 3
- 206010011891 Deafness neurosensory Diseases 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000012369 In process control Methods 0.000 description 3
- 208000027601 Inner ear disease Diseases 0.000 description 3
- 206010036626 Presbyacusis Diseases 0.000 description 3
- 208000009966 Sensorineural Hearing Loss Diseases 0.000 description 3
- 108700019146 Transgenes Proteins 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 108010077245 asparaginyl-proline Proteins 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 210000004544 dc2 Anatomy 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 210000000243 deiters cell Anatomy 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 210000004969 inflammatory cell Anatomy 0.000 description 3
- 238000004190 ion pair chromatography Methods 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 208000009800 presbycusis Diseases 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- 208000027491 vestibular disease Diseases 0.000 description 3
- GSCLWXDNIMNIJE-ZLUOBGJFSA-N Ala-Asp-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GSCLWXDNIMNIJE-ZLUOBGJFSA-N 0.000 description 2
- 108091023037 Aptamer Proteins 0.000 description 2
- WUQXMTITJLFXAU-JIOCBJNQSA-N Asn-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)N)N)O WUQXMTITJLFXAU-JIOCBJNQSA-N 0.000 description 2
- GHODABZPVZMWCE-FXQIFTODSA-N Asp-Glu-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O GHODABZPVZMWCE-FXQIFTODSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000000781 Conductive Hearing Loss Diseases 0.000 description 2
- 206010010280 Conductive deafness Diseases 0.000 description 2
- 206010011732 Cyst Diseases 0.000 description 2
- 208000003164 Diplopia Diseases 0.000 description 2
- BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 description 2
- 238000010867 Hoechst staining Methods 0.000 description 2
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 2
- 108010079364 N-glycylalanine Proteins 0.000 description 2
- 206010033109 Ototoxicity Diseases 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- ABWNZPOIUJMNKT-IXOXFDKPSA-N Thr-Phe-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O ABWNZPOIUJMNKT-IXOXFDKPSA-N 0.000 description 2
- 208000009205 Tinnitus Diseases 0.000 description 2
- 108090000704 Tubulin Proteins 0.000 description 2
- 102000004243 Tubulin Human genes 0.000 description 2
- 206010047348 Vertigo positional Diseases 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 108010060035 arginylproline Proteins 0.000 description 2
- 201000000691 benign paroxysmal positional nystagmus Diseases 0.000 description 2
- 208000001870 benign paroxysmal positional vertigo Diseases 0.000 description 2
- 239000007975 buffered saline Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 208000030251 communication disease Diseases 0.000 description 2
- 208000023563 conductive hearing loss disease Diseases 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000002716 delivery method Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 208000029444 double vision Diseases 0.000 description 2
- 230000004064 dysfunction Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 108010078144 glutaminyl-glycine Proteins 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- 208000034783 hypoesthesia Diseases 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 108010057821 leucylproline Proteins 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 208000018883 loss of balance Diseases 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 231100000262 ototoxicity Toxicity 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 108010077112 prolyl-proline Proteins 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 108010053725 prolylvaline Proteins 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 231100000879 sensorineural hearing loss Toxicity 0.000 description 2
- 208000023573 sensorineural hearing loss disease Diseases 0.000 description 2
- 230000001953 sensory effect Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 206010042772 syncope Diseases 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 231100000886 tinnitus Toxicity 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- 210000001213 vestibule labyrinth Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- BRPMXFSTKXXNHF-IUCAKERBSA-N (2s)-1-[2-[[(2s)-pyrrolidine-2-carbonyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)CNC(=O)[C@H]1NCCC1 BRPMXFSTKXXNHF-IUCAKERBSA-N 0.000 description 1
- LCSKNASZPVZHEG-UHFFFAOYSA-N 3,6-dimethyl-1,4-dioxane-2,5-dione;1,4-dioxane-2,5-dione Chemical group O=C1COC(=O)CO1.CC1OC(=O)C(C)OC1=O LCSKNASZPVZHEG-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241001655883 Adeno-associated virus - 1 Species 0.000 description 1
- 241000202702 Adeno-associated virus - 3 Species 0.000 description 1
- 241000580270 Adeno-associated virus - 4 Species 0.000 description 1
- 241000972680 Adeno-associated virus - 6 Species 0.000 description 1
- 241001164823 Adeno-associated virus - 7 Species 0.000 description 1
- FJVAQLJNTSUQPY-CIUDSAMLSA-N Ala-Ala-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN FJVAQLJNTSUQPY-CIUDSAMLSA-N 0.000 description 1
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 1
- LWUWMHIOBPTZBA-DCAQKATOSA-N Ala-Arg-Lys Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCCN)C(O)=O LWUWMHIOBPTZBA-DCAQKATOSA-N 0.000 description 1
- LBJYAILUMSUTAM-ZLUOBGJFSA-N Ala-Asn-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O LBJYAILUMSUTAM-ZLUOBGJFSA-N 0.000 description 1
- NHCPCLJZRSIDHS-ZLUOBGJFSA-N Ala-Asp-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O NHCPCLJZRSIDHS-ZLUOBGJFSA-N 0.000 description 1
- KIUYPHAMDKDICO-WHFBIAKZSA-N Ala-Asp-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O KIUYPHAMDKDICO-WHFBIAKZSA-N 0.000 description 1
- IKKVASZHTMKJIR-ZKWXMUAHSA-N Ala-Asp-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O IKKVASZHTMKJIR-ZKWXMUAHSA-N 0.000 description 1
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 1
- CCDFBRZVTDDJNM-GUBZILKMSA-N Ala-Leu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O CCDFBRZVTDDJNM-GUBZILKMSA-N 0.000 description 1
- XUCHENWTTBFODJ-FXQIFTODSA-N Ala-Met-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O XUCHENWTTBFODJ-FXQIFTODSA-N 0.000 description 1
- BHTBAVZSZCQZPT-GUBZILKMSA-N Ala-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N BHTBAVZSZCQZPT-GUBZILKMSA-N 0.000 description 1
- KLALXKYLOMZDQT-ZLUOBGJFSA-N Ala-Ser-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(N)=O KLALXKYLOMZDQT-ZLUOBGJFSA-N 0.000 description 1
- YCTIYBUTCKNOTI-UWJYBYFXSA-N Ala-Tyr-Asp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N YCTIYBUTCKNOTI-UWJYBYFXSA-N 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 208000024985 Alport syndrome Diseases 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- BVBKBQRPOJFCQM-DCAQKATOSA-N Arg-Asn-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BVBKBQRPOJFCQM-DCAQKATOSA-N 0.000 description 1
- OCOZPTHLDVSFCZ-BPUTZDHNSA-N Arg-Asn-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N OCOZPTHLDVSFCZ-BPUTZDHNSA-N 0.000 description 1
- ALOVURZCXKYKJC-NAKRPEOUSA-N Arg-Asp-Gln-Ser Chemical compound N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O ALOVURZCXKYKJC-NAKRPEOUSA-N 0.000 description 1
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 1
- VXXHDZKEQNGXNU-QXEWZRGKSA-N Arg-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N VXXHDZKEQNGXNU-QXEWZRGKSA-N 0.000 description 1
- KBBKCNHWCDJPGN-GUBZILKMSA-N Arg-Gln-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O KBBKCNHWCDJPGN-GUBZILKMSA-N 0.000 description 1
- VNFWDYWTSHFRRG-SRVKXCTJSA-N Arg-Gln-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O VNFWDYWTSHFRRG-SRVKXCTJSA-N 0.000 description 1
- BQBPFMNVOWDLHO-XIRDDKMYSA-N Arg-Gln-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N BQBPFMNVOWDLHO-XIRDDKMYSA-N 0.000 description 1
- UHFUZWSZQKMDSX-DCAQKATOSA-N Arg-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UHFUZWSZQKMDSX-DCAQKATOSA-N 0.000 description 1
- UZGFHWIJWPUPOH-IHRRRGAJSA-N Arg-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UZGFHWIJWPUPOH-IHRRRGAJSA-N 0.000 description 1
- IGFJVXOATGZTHD-UHFFFAOYSA-N Arg-Phe-His Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccccc1)C(=O)NC(Cc2c[nH]cn2)C(=O)O IGFJVXOATGZTHD-UHFFFAOYSA-N 0.000 description 1
- UULLJGQFCDXVTQ-CYDGBPFRSA-N Arg-Pro-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UULLJGQFCDXVTQ-CYDGBPFRSA-N 0.000 description 1
- UZSQXCMNUPKLCC-FJXKBIBVSA-N Arg-Thr-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UZSQXCMNUPKLCC-FJXKBIBVSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- CPTXATAOUQJQRO-GUBZILKMSA-N Arg-Val-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O CPTXATAOUQJQRO-GUBZILKMSA-N 0.000 description 1
- HUZGPXBILPMCHM-IHRRRGAJSA-N Asn-Arg-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HUZGPXBILPMCHM-IHRRRGAJSA-N 0.000 description 1
- PCKRJVZAQZWNKM-WHFBIAKZSA-N Asn-Asn-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O PCKRJVZAQZWNKM-WHFBIAKZSA-N 0.000 description 1
- AYZAWXAPBAYCHO-CIUDSAMLSA-N Asn-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N AYZAWXAPBAYCHO-CIUDSAMLSA-N 0.000 description 1
- NVGWESORMHFISY-SRVKXCTJSA-N Asn-Asn-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O NVGWESORMHFISY-SRVKXCTJSA-N 0.000 description 1
- JRVABKHPWDRUJF-UBHSHLNASA-N Asn-Asn-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N JRVABKHPWDRUJF-UBHSHLNASA-N 0.000 description 1
- GNKVBRYFXYWXAB-WDSKDSINSA-N Asn-Glu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O GNKVBRYFXYWXAB-WDSKDSINSA-N 0.000 description 1
- OOWSBIOUKIUWLO-RCOVLWMOSA-N Asn-Gly-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O OOWSBIOUKIUWLO-RCOVLWMOSA-N 0.000 description 1
- PHJPKNUWWHRAOC-PEFMBERDSA-N Asn-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N PHJPKNUWWHRAOC-PEFMBERDSA-N 0.000 description 1
- COWITDLVHMZSIW-CIUDSAMLSA-N Asn-Lys-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O COWITDLVHMZSIW-CIUDSAMLSA-N 0.000 description 1
- YUOXLJYVSZYPBJ-CIUDSAMLSA-N Asn-Pro-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O YUOXLJYVSZYPBJ-CIUDSAMLSA-N 0.000 description 1
- GZXOUBTUAUAVHD-ACZMJKKPSA-N Asn-Ser-Glu Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GZXOUBTUAUAVHD-ACZMJKKPSA-N 0.000 description 1
- QUMKPKWYDVMGNT-NUMRIWBASA-N Asn-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QUMKPKWYDVMGNT-NUMRIWBASA-N 0.000 description 1
- JZLFYAAGGYMRIK-BYULHYEWSA-N Asn-Val-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O JZLFYAAGGYMRIK-BYULHYEWSA-N 0.000 description 1
- XEDQMTWEYFBOIK-ACZMJKKPSA-N Asp-Ala-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O XEDQMTWEYFBOIK-ACZMJKKPSA-N 0.000 description 1
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 1
- VBVKSAFJPVXMFJ-CIUDSAMLSA-N Asp-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N VBVKSAFJPVXMFJ-CIUDSAMLSA-N 0.000 description 1
- UFAQGGZUXVLONR-AVGNSLFASA-N Asp-Gln-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)O)N)O UFAQGGZUXVLONR-AVGNSLFASA-N 0.000 description 1
- ZSVJVIOVABDTTL-YUMQZZPRSA-N Asp-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)O)N ZSVJVIOVABDTTL-YUMQZZPRSA-N 0.000 description 1
- SVABRQFIHCSNCI-FOHZUACHSA-N Asp-Gly-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O SVABRQFIHCSNCI-FOHZUACHSA-N 0.000 description 1
- PGUYEUCYVNZGGV-QWRGUYRKSA-N Asp-Gly-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PGUYEUCYVNZGGV-QWRGUYRKSA-N 0.000 description 1
- USNJAPJZSGTTPX-XVSYOHENSA-N Asp-Phe-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O USNJAPJZSGTTPX-XVSYOHENSA-N 0.000 description 1
- KGHLGJAXYSVNJP-WHFBIAKZSA-N Asp-Ser-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O KGHLGJAXYSVNJP-WHFBIAKZSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 1
- JSNWZMFSLIWAHS-HJGDQZAQSA-N Asp-Thr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O JSNWZMFSLIWAHS-HJGDQZAQSA-N 0.000 description 1
- LLRJPYJQNBMOOO-QEJZJMRPSA-N Asp-Trp-Gln Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N LLRJPYJQNBMOOO-QEJZJMRPSA-N 0.000 description 1
- QPDUWAUSSWGJSB-NGZCFLSTSA-N Asp-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N QPDUWAUSSWGJSB-NGZCFLSTSA-N 0.000 description 1
- 206010003497 Asphyxia Diseases 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091016585 CD44 antigen Proteins 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 208000026091 Congenital hearing disease Diseases 0.000 description 1
- XIZWKXATMJODQW-KKUMJFAQSA-N Cys-His-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CS)N XIZWKXATMJODQW-KKUMJFAQSA-N 0.000 description 1
- XLLSMEFANRROJE-GUBZILKMSA-N Cys-Leu-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CS)N XLLSMEFANRROJE-GUBZILKMSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 206010011882 Deafness congenital Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 206010016717 Fistula Diseases 0.000 description 1
- 108010046649 GDNP peptide Proteins 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 241000699694 Gerbillinae Species 0.000 description 1
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 1
- JSYULGSPLTZDHM-NRPADANISA-N Gln-Ala-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O JSYULGSPLTZDHM-NRPADANISA-N 0.000 description 1
- IVCOYUURLWQDJQ-LPEHRKFASA-N Gln-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O IVCOYUURLWQDJQ-LPEHRKFASA-N 0.000 description 1
- HVQCEQTUSWWFOS-WDSKDSINSA-N Gln-Gly-Cys Chemical compound C(CC(=O)N)[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N HVQCEQTUSWWFOS-WDSKDSINSA-N 0.000 description 1
- SMLDOQHTOAAFJQ-WDSKDSINSA-N Gln-Gly-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SMLDOQHTOAAFJQ-WDSKDSINSA-N 0.000 description 1
- ZBKUIQNCRIYVGH-SDDRHHMPSA-N Gln-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N ZBKUIQNCRIYVGH-SDDRHHMPSA-N 0.000 description 1
- FALJZCPMTGJOHX-SRVKXCTJSA-N Gln-Met-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O FALJZCPMTGJOHX-SRVKXCTJSA-N 0.000 description 1
- UESYBOXFJWJVSB-AVGNSLFASA-N Gln-Phe-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O UESYBOXFJWJVSB-AVGNSLFASA-N 0.000 description 1
- XQDGOJPVMSWZSO-SRVKXCTJSA-N Gln-Pro-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N XQDGOJPVMSWZSO-SRVKXCTJSA-N 0.000 description 1
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-N 0.000 description 1
- PBEQPAZRHDVJQI-SRVKXCTJSA-N Glu-Arg-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)O)N PBEQPAZRHDVJQI-SRVKXCTJSA-N 0.000 description 1
- RDDSZZJOKDVPAE-ACZMJKKPSA-N Glu-Asn-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RDDSZZJOKDVPAE-ACZMJKKPSA-N 0.000 description 1
- SBCYJMOOHUDWDA-NUMRIWBASA-N Glu-Asp-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SBCYJMOOHUDWDA-NUMRIWBASA-N 0.000 description 1
- CUXJIASLBRJOFV-LAEOZQHASA-N Glu-Gly-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CUXJIASLBRJOFV-LAEOZQHASA-N 0.000 description 1
- QIQABBIDHGQXGA-ZPFDUUQYSA-N Glu-Ile-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QIQABBIDHGQXGA-ZPFDUUQYSA-N 0.000 description 1
- DNPCBMNFQVTHMA-DCAQKATOSA-N Glu-Leu-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O DNPCBMNFQVTHMA-DCAQKATOSA-N 0.000 description 1
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- GWCRIHNSVMOBEQ-BQBZGAKWSA-N Gly-Arg-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O GWCRIHNSVMOBEQ-BQBZGAKWSA-N 0.000 description 1
- DWUKOTKSTDWGAE-BQBZGAKWSA-N Gly-Asn-Arg Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DWUKOTKSTDWGAE-BQBZGAKWSA-N 0.000 description 1
- KQDMENMTYNBWMR-WHFBIAKZSA-N Gly-Asp-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KQDMENMTYNBWMR-WHFBIAKZSA-N 0.000 description 1
- PABFFPWEJMEVEC-JGVFFNPUSA-N Gly-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)CN)C(=O)O PABFFPWEJMEVEC-JGVFFNPUSA-N 0.000 description 1
- QPDUVFSVVAOUHE-XVKPBYJWSA-N Gly-Gln-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)CN)C(O)=O QPDUVFSVVAOUHE-XVKPBYJWSA-N 0.000 description 1
- YTSVAIMKVLZUDU-YUMQZZPRSA-N Gly-Leu-Asp Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O YTSVAIMKVLZUDU-YUMQZZPRSA-N 0.000 description 1
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 1
- FXLVSYVJDPCIHH-STQMWFEESA-N Gly-Phe-Arg Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FXLVSYVJDPCIHH-STQMWFEESA-N 0.000 description 1
- SSFWXSNOKDZNHY-QXEWZRGKSA-N Gly-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN SSFWXSNOKDZNHY-QXEWZRGKSA-N 0.000 description 1
- ZLCLYFGMKFCDCN-XPUUQOCRSA-N Gly-Ser-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CO)NC(=O)CN)C(O)=O ZLCLYFGMKFCDCN-XPUUQOCRSA-N 0.000 description 1
- YXTFLTJYLIAZQG-FJXKBIBVSA-N Gly-Thr-Arg Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YXTFLTJYLIAZQG-FJXKBIBVSA-N 0.000 description 1
- FKESCSGWBPUTPN-FOHZUACHSA-N Gly-Thr-Asn Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O FKESCSGWBPUTPN-FOHZUACHSA-N 0.000 description 1
- PNUFMLXHOLFRLD-KBPBESRZSA-N Gly-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 PNUFMLXHOLFRLD-KBPBESRZSA-N 0.000 description 1
- 102000008055 Heparan Sulfate Proteoglycans Human genes 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- BDHUXUFYNUOUIT-SRVKXCTJSA-N His-Asp-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BDHUXUFYNUOUIT-SRVKXCTJSA-N 0.000 description 1
- JENKOCSDMSVWPY-SRVKXCTJSA-N His-Leu-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O JENKOCSDMSVWPY-SRVKXCTJSA-N 0.000 description 1
- LNDVNHOSZQPJGI-AVGNSLFASA-N His-Pro-Pro Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CN=CN1 LNDVNHOSZQPJGI-AVGNSLFASA-N 0.000 description 1
- PLCAEMGSYOYIPP-GUBZILKMSA-N His-Ser-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 PLCAEMGSYOYIPP-GUBZILKMSA-N 0.000 description 1
- GIRSNERMXCMDBO-GARJFASQSA-N His-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O GIRSNERMXCMDBO-GARJFASQSA-N 0.000 description 1
- 101000808906 Homo sapiens AT-rich interactive domain-containing protein 3B Proteins 0.000 description 1
- 101000574396 Homo sapiens Protein phosphatase 1K, mitochondrial Proteins 0.000 description 1
- 101000788505 Homo sapiens TBC1 domain family member 24 Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- HTDRTKMNJRRYOJ-SIUGBPQLSA-N Ile-Gln-Tyr Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HTDRTKMNJRRYOJ-SIUGBPQLSA-N 0.000 description 1
- DFJJAVZIHDFOGQ-MNXVOIDGSA-N Ile-Glu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DFJJAVZIHDFOGQ-MNXVOIDGSA-N 0.000 description 1
- LEHPJMKVGFPSSP-ZQINRCPSSA-N Ile-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)[C@@H](C)CC)C(O)=O)=CNC2=C1 LEHPJMKVGFPSSP-ZQINRCPSSA-N 0.000 description 1
- UAELWXJFLZBKQS-WHOFXGATSA-N Ile-Phe-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)NCC(O)=O UAELWXJFLZBKQS-WHOFXGATSA-N 0.000 description 1
- BJECXJHLUJXPJQ-PYJNHQTQSA-N Ile-Pro-His Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N BJECXJHLUJXPJQ-PYJNHQTQSA-N 0.000 description 1
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- MDVZJYGNAGLPGJ-KKUMJFAQSA-N Leu-Asn-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MDVZJYGNAGLPGJ-KKUMJFAQSA-N 0.000 description 1
- VPKIQULSKFVCSM-SRVKXCTJSA-N Leu-Gln-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VPKIQULSKFVCSM-SRVKXCTJSA-N 0.000 description 1
- KGCLIYGPQXUNLO-IUCAKERBSA-N Leu-Gly-Glu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O KGCLIYGPQXUNLO-IUCAKERBSA-N 0.000 description 1
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 1
- USLNHQZCDQJBOV-ZPFDUUQYSA-N Leu-Ile-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O USLNHQZCDQJBOV-ZPFDUUQYSA-N 0.000 description 1
- HNDWYLYAYNBWMP-AJNGGQMLSA-N Leu-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N HNDWYLYAYNBWMP-AJNGGQMLSA-N 0.000 description 1
- ARRIJPQRBWRNLT-DCAQKATOSA-N Leu-Met-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(=O)N)C(=O)O)N ARRIJPQRBWRNLT-DCAQKATOSA-N 0.000 description 1
- BMVFXOQHDQZAQU-DCAQKATOSA-N Leu-Pro-Asp Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N BMVFXOQHDQZAQU-DCAQKATOSA-N 0.000 description 1
- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 1
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 1
- IDGZVZJLYFTXSL-DCAQKATOSA-N Leu-Ser-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IDGZVZJLYFTXSL-DCAQKATOSA-N 0.000 description 1
- QWWPYKKLXWOITQ-VOAKCMCISA-N Leu-Thr-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QWWPYKKLXWOITQ-VOAKCMCISA-N 0.000 description 1
- YIRIDPUGZKHMHT-ACRUOGEOSA-N Leu-Tyr-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YIRIDPUGZKHMHT-ACRUOGEOSA-N 0.000 description 1
- XZNJZXJZBMBGGS-NHCYSSNCSA-N Leu-Val-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XZNJZXJZBMBGGS-NHCYSSNCSA-N 0.000 description 1
- 101710174256 Leucine-rich repeat-containing G-protein coupled receptor 5 Proteins 0.000 description 1
- XFIHDSBIPWEYJJ-YUMQZZPRSA-N Lys-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN XFIHDSBIPWEYJJ-YUMQZZPRSA-N 0.000 description 1
- NLOZZWJNIKKYSC-WDSOQIARSA-N Lys-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CCCCN)C(O)=O)=CNC2=C1 NLOZZWJNIKKYSC-WDSOQIARSA-N 0.000 description 1
- QUYCUALODHJQLK-CIUDSAMLSA-N Lys-Asp-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O QUYCUALODHJQLK-CIUDSAMLSA-N 0.000 description 1
- ULUQBUKAPDUKOC-GVXVVHGQSA-N Lys-Glu-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O ULUQBUKAPDUKOC-GVXVVHGQSA-N 0.000 description 1
- LCMWVZLBCUVDAZ-IUCAKERBSA-N Lys-Gly-Glu Chemical compound [NH3+]CCCC[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CCC([O-])=O LCMWVZLBCUVDAZ-IUCAKERBSA-N 0.000 description 1
- XIZQPFCRXLUNMK-BZSNNMDCSA-N Lys-Leu-Phe Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCCCN)N XIZQPFCRXLUNMK-BZSNNMDCSA-N 0.000 description 1
- IPTUBUUIFRZMJK-ACRUOGEOSA-N Lys-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 IPTUBUUIFRZMJK-ACRUOGEOSA-N 0.000 description 1
- BOJYMMBYBNOOGG-DCAQKATOSA-N Lys-Pro-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O BOJYMMBYBNOOGG-DCAQKATOSA-N 0.000 description 1
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 1
- RMOKGALPSPOYKE-KATARQTJSA-N Lys-Thr-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O RMOKGALPSPOYKE-KATARQTJSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-L Malonate Chemical compound [O-]C(=O)CC([O-])=O OFOBLEOULBTSOW-UHFFFAOYSA-L 0.000 description 1
- 208000027530 Meniere disease Diseases 0.000 description 1
- YRAWWKUTNBILNT-FXQIFTODSA-N Met-Ala-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YRAWWKUTNBILNT-FXQIFTODSA-N 0.000 description 1
- UZWMJZSOXGOVIN-LURJTMIESA-N Met-Gly-Gly Chemical compound CSCC[C@H](N)C(=O)NCC(=O)NCC(O)=O UZWMJZSOXGOVIN-LURJTMIESA-N 0.000 description 1
- 208000008719 Mixed Conductive-Sensorineural Hearing Loss Diseases 0.000 description 1
- 241000282339 Mustela Species 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- UHRNIXJAGGLKHP-DLOVCJGASA-N Phe-Ala-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O UHRNIXJAGGLKHP-DLOVCJGASA-N 0.000 description 1
- DJPXNKUDJKGQEE-BZSNNMDCSA-N Phe-Asp-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DJPXNKUDJKGQEE-BZSNNMDCSA-N 0.000 description 1
- UNLYPPYNDXHGDG-IHRRRGAJSA-N Phe-Gln-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UNLYPPYNDXHGDG-IHRRRGAJSA-N 0.000 description 1
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 1
- APJPXSFJBMMOLW-KBPBESRZSA-N Phe-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 APJPXSFJBMMOLW-KBPBESRZSA-N 0.000 description 1
- WFHRXJOZEXUKLV-IRXDYDNUSA-N Phe-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 WFHRXJOZEXUKLV-IRXDYDNUSA-N 0.000 description 1
- BEEVXUYVEHXWRQ-YESZJQIVSA-N Phe-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC3=CC=CC=C3)N)C(=O)O BEEVXUYVEHXWRQ-YESZJQIVSA-N 0.000 description 1
- MYQCCQSMKNCNKY-KKUMJFAQSA-N Phe-His-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CO)C(=O)O)N MYQCCQSMKNCNKY-KKUMJFAQSA-N 0.000 description 1
- BYAIIACBWBOJCU-URLPEUOOSA-N Phe-Ile-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BYAIIACBWBOJCU-URLPEUOOSA-N 0.000 description 1
- JLLJTMHNXQTMCK-UBHSHLNASA-N Phe-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 JLLJTMHNXQTMCK-UBHSHLNASA-N 0.000 description 1
- RAGOJJCBGXARPO-XVSYOHENSA-N Phe-Thr-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RAGOJJCBGXARPO-XVSYOHENSA-N 0.000 description 1
- YFNOUBWUIIJQHF-LPEHRKFASA-N Pro-Asp-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)O)C(=O)N2CCC[C@@H]2C(=O)O YFNOUBWUIIJQHF-LPEHRKFASA-N 0.000 description 1
- YKQNVTOIYFQMLW-IHRRRGAJSA-N Pro-Cys-Tyr Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H]1NCCC1)C1=CC=C(O)C=C1 YKQNVTOIYFQMLW-IHRRRGAJSA-N 0.000 description 1
- WGAQWMRJUFQXMF-ZPFDUUQYSA-N Pro-Gln-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WGAQWMRJUFQXMF-ZPFDUUQYSA-N 0.000 description 1
- DIFXZGPHVCIVSQ-CIUDSAMLSA-N Pro-Gln-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O DIFXZGPHVCIVSQ-CIUDSAMLSA-N 0.000 description 1
- FEVDNIBDCRKMER-IUCAKERBSA-N Pro-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@@H]1CCCN1 FEVDNIBDCRKMER-IUCAKERBSA-N 0.000 description 1
- BRJGUPWVFXKBQI-XUXIUFHCSA-N Pro-Leu-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BRJGUPWVFXKBQI-XUXIUFHCSA-N 0.000 description 1
- JUJCUYWRJMFJJF-AVGNSLFASA-N Pro-Lys-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H]1CCCN1 JUJCUYWRJMFJJF-AVGNSLFASA-N 0.000 description 1
- VGVCNKSUVSZEIE-IHRRRGAJSA-N Pro-Phe-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O VGVCNKSUVSZEIE-IHRRRGAJSA-N 0.000 description 1
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 1
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-N 0.000 description 1
- VVAWNPIOYXAMAL-KJEVXHAQSA-N Pro-Thr-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VVAWNPIOYXAMAL-KJEVXHAQSA-N 0.000 description 1
- XDKKMRPRRCOELJ-GUBZILKMSA-N Pro-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 XDKKMRPRRCOELJ-GUBZILKMSA-N 0.000 description 1
- IMNVAOPEMFDAQD-NHCYSSNCSA-N Pro-Val-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IMNVAOPEMFDAQD-NHCYSSNCSA-N 0.000 description 1
- IIRBTQHFVNGPMQ-AVGNSLFASA-N Pro-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@@H]1CCCN1 IIRBTQHFVNGPMQ-AVGNSLFASA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 108010079005 RDV peptide Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- DWUIECHTAMYEFL-XVYDVKMFSA-N Ser-Ala-His Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 DWUIECHTAMYEFL-XVYDVKMFSA-N 0.000 description 1
- HQTKVSCNCDLXSX-BQBZGAKWSA-N Ser-Arg-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O HQTKVSCNCDLXSX-BQBZGAKWSA-N 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 1
- UFKPDBLKLOBMRH-XHNCKOQMSA-N Ser-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N)C(=O)O UFKPDBLKLOBMRH-XHNCKOQMSA-N 0.000 description 1
- WBINSDOPZHQPPM-AVGNSLFASA-N Ser-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N)O WBINSDOPZHQPPM-AVGNSLFASA-N 0.000 description 1
- BPMRXBZYPGYPJN-WHFBIAKZSA-N Ser-Gly-Asn Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O BPMRXBZYPGYPJN-WHFBIAKZSA-N 0.000 description 1
- GZFAWAQTEYDKII-YUMQZZPRSA-N Ser-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO GZFAWAQTEYDKII-YUMQZZPRSA-N 0.000 description 1
- CLKKNZQUQMZDGD-SRVKXCTJSA-N Ser-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CC1=CN=CN1 CLKKNZQUQMZDGD-SRVKXCTJSA-N 0.000 description 1
- NLOAIFSWUUFQFR-CIUDSAMLSA-N Ser-Leu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O NLOAIFSWUUFQFR-CIUDSAMLSA-N 0.000 description 1
- XKFJENWJGHMDLI-QWRGUYRKSA-N Ser-Phe-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O XKFJENWJGHMDLI-QWRGUYRKSA-N 0.000 description 1
- ZKBKUWQVDWWSRI-BZSNNMDCSA-N Ser-Phe-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKBKUWQVDWWSRI-BZSNNMDCSA-N 0.000 description 1
- NUEHQDHDLDXCRU-GUBZILKMSA-N Ser-Pro-Arg Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NUEHQDHDLDXCRU-GUBZILKMSA-N 0.000 description 1
- FKYWFUYPVKLJLP-DCAQKATOSA-N Ser-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FKYWFUYPVKLJLP-DCAQKATOSA-N 0.000 description 1
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 1
- ZSDXEKUKQAKZFE-XAVMHZPKSA-N Ser-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N)O ZSDXEKUKQAKZFE-XAVMHZPKSA-N 0.000 description 1
- PIQRHJQWEPWFJG-UWJYBYFXSA-N Ser-Tyr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PIQRHJQWEPWFJG-UWJYBYFXSA-N 0.000 description 1
- BEBVVQPDSHHWQL-NRPADANISA-N Ser-Val-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O BEBVVQPDSHHWQL-NRPADANISA-N 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 108090000054 Syndecan-2 Proteins 0.000 description 1
- 108010008038 Synthetic Vaccines Proteins 0.000 description 1
- 102100025233 TBC1 domain family member 24 Human genes 0.000 description 1
- LVHHEVGYAZGXDE-KDXUFGMBSA-N Thr-Ala-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(=O)O)N)O LVHHEVGYAZGXDE-KDXUFGMBSA-N 0.000 description 1
- WFUAUEQXPVNAEF-ZJDVBMNYSA-N Thr-Arg-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)O)C(O)=O)CCCN=C(N)N WFUAUEQXPVNAEF-ZJDVBMNYSA-N 0.000 description 1
- GCXFWAZRHBRYEM-NUMRIWBASA-N Thr-Gln-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O GCXFWAZRHBRYEM-NUMRIWBASA-N 0.000 description 1
- GKWNLDNXMMLRMC-GLLZPBPUSA-N Thr-Glu-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O GKWNLDNXMMLRMC-GLLZPBPUSA-N 0.000 description 1
- SLUWOCTZVGMURC-BFHQHQDPSA-N Thr-Gly-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O SLUWOCTZVGMURC-BFHQHQDPSA-N 0.000 description 1
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 1
- KKPOGALELPLJTL-MEYUZBJRSA-N Thr-Lys-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 KKPOGALELPLJTL-MEYUZBJRSA-N 0.000 description 1
- UJQVSMNQMQHVRY-KZVJFYERSA-N Thr-Met-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O UJQVSMNQMQHVRY-KZVJFYERSA-N 0.000 description 1
- STUAPCLEDMKXKL-LKXGYXEUSA-N Thr-Ser-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O STUAPCLEDMKXKL-LKXGYXEUSA-N 0.000 description 1
- RVMNUBQWPVOUKH-HEIBUPTGSA-N Thr-Ser-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMNUBQWPVOUKH-HEIBUPTGSA-N 0.000 description 1
- QYDKSNXSBXZPFK-ZJDVBMNYSA-N Thr-Thr-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYDKSNXSBXZPFK-ZJDVBMNYSA-N 0.000 description 1
- AAZOYLQUEQRUMZ-GSSVUCPTSA-N Thr-Thr-Asn Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O AAZOYLQUEQRUMZ-GSSVUCPTSA-N 0.000 description 1
- MFMGPEKYBXFIRF-SUSMZKCASA-N Thr-Thr-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MFMGPEKYBXFIRF-SUSMZKCASA-N 0.000 description 1
- NHQVWACSJZJCGJ-FLBSBUHZSA-N Thr-Thr-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NHQVWACSJZJCGJ-FLBSBUHZSA-N 0.000 description 1
- ZMYCLHFLHRVOEA-HEIBUPTGSA-N Thr-Thr-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ZMYCLHFLHRVOEA-HEIBUPTGSA-N 0.000 description 1
- NMCBVGFGWSIGSB-NUTKFTJISA-N Trp-Ala-Leu Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N NMCBVGFGWSIGSB-NUTKFTJISA-N 0.000 description 1
- HYVLNORXQGKONN-NUTKFTJISA-N Trp-Ala-Lys Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O)=CNC2=C1 HYVLNORXQGKONN-NUTKFTJISA-N 0.000 description 1
- WVHUFSCKCBQKJW-HKUYNNGSSA-N Trp-Gly-Tyr Chemical compound C([C@H](NC(=O)CNC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=C(O)C=C1 WVHUFSCKCBQKJW-HKUYNNGSSA-N 0.000 description 1
- YRSOERSDNRSCBC-XIRDDKMYSA-N Trp-His-Cys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CN=CN3)C(=O)N[C@@H](CS)C(=O)O)N YRSOERSDNRSCBC-XIRDDKMYSA-N 0.000 description 1
- VPRHDRKAPYZMHL-SZMVWBNQSA-N Trp-Leu-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 VPRHDRKAPYZMHL-SZMVWBNQSA-N 0.000 description 1
- NWQCKAPDGQMZQN-IHPCNDPISA-N Trp-Lys-Leu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O NWQCKAPDGQMZQN-IHPCNDPISA-N 0.000 description 1
- RERRMBXDSFMBQE-ZFWWWQNUSA-N Trp-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N RERRMBXDSFMBQE-ZFWWWQNUSA-N 0.000 description 1
- CYDVHRFXDMDMGX-KKUMJFAQSA-N Tyr-Asn-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O CYDVHRFXDMDMGX-KKUMJFAQSA-N 0.000 description 1
- NMKJPMCEKQHRPD-IRXDYDNUSA-N Tyr-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NMKJPMCEKQHRPD-IRXDYDNUSA-N 0.000 description 1
- NKUGCYDFQKFVOJ-JYJNAYRXSA-N Tyr-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NKUGCYDFQKFVOJ-JYJNAYRXSA-N 0.000 description 1
- DWAMXBFJNZIHMC-KBPBESRZSA-N Tyr-Leu-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O DWAMXBFJNZIHMC-KBPBESRZSA-N 0.000 description 1
- PRONOHBTMLNXCZ-BZSNNMDCSA-N Tyr-Leu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 PRONOHBTMLNXCZ-BZSNNMDCSA-N 0.000 description 1
- DMWNPLOERDAHSY-MEYUZBJRSA-N Tyr-Leu-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DMWNPLOERDAHSY-MEYUZBJRSA-N 0.000 description 1
- BYAKMYBZADCNMN-JYJNAYRXSA-N Tyr-Lys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYAKMYBZADCNMN-JYJNAYRXSA-N 0.000 description 1
- WURLIFOWSMBUAR-SLFFLAALSA-N Tyr-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)C(=O)O WURLIFOWSMBUAR-SLFFLAALSA-N 0.000 description 1
- LUMQYLVYUIRHHU-YJRXYDGGSA-N Tyr-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LUMQYLVYUIRHHU-YJRXYDGGSA-N 0.000 description 1
- PLVVHGFEMSDRET-IHPCNDPISA-N Tyr-Ser-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC3=CC=C(C=C3)O)N PLVVHGFEMSDRET-IHPCNDPISA-N 0.000 description 1
- LDKDSFQSEUOCOO-RPTUDFQQSA-N Tyr-Thr-Phe Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LDKDSFQSEUOCOO-RPTUDFQQSA-N 0.000 description 1
- 208000014769 Usher Syndromes Diseases 0.000 description 1
- ZMDCGGKHRKNWKD-LAEOZQHASA-N Val-Asn-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZMDCGGKHRKNWKD-LAEOZQHASA-N 0.000 description 1
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 1
- OVLIFGQSBSNGHY-KKHAAJSZSA-N Val-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N)O OVLIFGQSBSNGHY-KKHAAJSZSA-N 0.000 description 1
- UZDHNIJRRTUKKC-DLOVCJGASA-N Val-Gln-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N UZDHNIJRRTUKKC-DLOVCJGASA-N 0.000 description 1
- SZTTYWIUCGSURQ-AUTRQRHGSA-N Val-Glu-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SZTTYWIUCGSURQ-AUTRQRHGSA-N 0.000 description 1
- WDIGUPHXPBMODF-UMNHJUIQSA-N Val-Glu-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N WDIGUPHXPBMODF-UMNHJUIQSA-N 0.000 description 1
- NXRAUQGGHPCJIB-RCOVLWMOSA-N Val-Gly-Asn Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O NXRAUQGGHPCJIB-RCOVLWMOSA-N 0.000 description 1
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 1
- ZHQWPWQNVRCXAX-XQQFMLRXSA-N Val-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZHQWPWQNVRCXAX-XQQFMLRXSA-N 0.000 description 1
- MGVYZTPLGXPVQB-CYDGBPFRSA-N Val-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](C(C)C)N MGVYZTPLGXPVQB-CYDGBPFRSA-N 0.000 description 1
- ZEBRMWPTJNHXAJ-JYJNAYRXSA-N Val-Phe-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)O)N ZEBRMWPTJNHXAJ-JYJNAYRXSA-N 0.000 description 1
- XBJKAZATRJBDCU-GUBZILKMSA-N Val-Pro-Ala Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O XBJKAZATRJBDCU-GUBZILKMSA-N 0.000 description 1
- GQMNEJMFMCJJTD-NHCYSSNCSA-N Val-Pro-Gln Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O GQMNEJMFMCJJTD-NHCYSSNCSA-N 0.000 description 1
- NGXQOQNXSGOYOI-BQFCYCMXSA-N Val-Trp-Gln Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 NGXQOQNXSGOYOI-BQFCYCMXSA-N 0.000 description 1
- 208000014070 Vestibular schwannoma Diseases 0.000 description 1
- 208000026724 Waardenburg syndrome Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 208000004064 acoustic neuroma Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 210000003984 auditory pathway Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 210000000238 cell of claudius Anatomy 0.000 description 1
- 208000005675 central hearing loss Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000000306 component Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 210000003792 cranial nerve Anatomy 0.000 description 1
- 230000037029 cross reaction Effects 0.000 description 1
- 231100000895 deafness Toxicity 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 210000000883 ear external Anatomy 0.000 description 1
- 210000000959 ear middle Anatomy 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000003060 endolymph Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 210000000256 facial nerve Anatomy 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000010685 fatty oil Substances 0.000 description 1
- 230000003890 fistula Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000011990 functional testing Methods 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- HPAIKDPJURGQLN-UHFFFAOYSA-N glycyl-L-histidyl-L-phenylalanine Natural products C=1C=CC=CC=1CC(C(O)=O)NC(=O)C(NC(=O)CN)CC1=CN=CN1 HPAIKDPJURGQLN-UHFFFAOYSA-N 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010059898 glycyl-tyrosyl-lysine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 208000003215 hereditary nephritis Diseases 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 201000006790 nonsyndromic deafness Diseases 0.000 description 1
- 231100000862 numbness Toxicity 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000004789 organ system Anatomy 0.000 description 1
- 239000006179 pH buffering agent Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 210000004049 perilymph Anatomy 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 1
- 108010024654 phenylalanyl-prolyl-alanine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 210000000608 photoreceptor cell Anatomy 0.000 description 1
- 230000006461 physiological response Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920003196 poly(1,3-dioxolane) Polymers 0.000 description 1
- 238000002459 porosimetry Methods 0.000 description 1
- 238000013105 post hoc analysis Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108700042769 prolyl-leucyl-glycine Proteins 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 230000009145 protein modification Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000013608 rAAV vector Substances 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000001525 retina Anatomy 0.000 description 1
- 210000003569 retinal bipolar cell Anatomy 0.000 description 1
- 230000002207 retinal effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000001079 scala tympani Anatomy 0.000 description 1
- 210000001605 scala vestibuli Anatomy 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000004416 surface enhanced Raman spectroscopy Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 210000003582 temporal bone Anatomy 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 108010015666 tryptophyl-leucyl-glutamic acid Proteins 0.000 description 1
- 108010045269 tryptophyltryptophan Proteins 0.000 description 1
- 210000003454 tympanic membrane Anatomy 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 210000002905 vestibular aqueduct Anatomy 0.000 description 1
- 201000000200 vestibular neuronitis Diseases 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- BPICBUSOMSTKRF-UHFFFAOYSA-N xylazine Chemical compound CC1=CC=CC(C)=C1NC1=NCCCS1 BPICBUSOMSTKRF-UHFFFAOYSA-N 0.000 description 1
- 229960001600 xylazine Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- 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
- A61K48/0075—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 delivery route, e.g. oral, subcutaneous
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/16—Otologicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/14011—Parvoviridae
- C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
- C12N2750/14122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/14011—Parvoviridae
- C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
- C12N2750/14141—Use of virus, viral particle or viral elements as a vector
- C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Pharmacology & Pharmacy (AREA)
- Molecular Biology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Virology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本文提供了腺相关病毒及其用于治疗或预防影响受试者内耳的病症的用途。
Description
优先权相关申请
本申请要求于2018年12月21日提交的美国临时专利申请No.62/784306的权益,该申请通过引用在此整体并入。
背景技术
听力损失是影响现今世界人口的最常见的失能之一。根据国家健康和营养检查调查(the National Health and Nutritional Examination Survey),近三分之二的70岁及以上的美国成年人患有听力损失。听力损失与内耳毛细胞损伤有关。内耳基因疗法是一种很有前途的治疗方法,它可以潜在地预防和逆转毛细胞损伤。虽然腺相关病毒载体(AAV)介导的内耳基因疗法已应用于遗传性听力损失的动物模型以改善听觉功能,但某些内耳毛细胞的感染效率较低。此外,传统AAVs对内耳中的支持细胞的感染效率也较低。为使内耳基因疗法有效地治疗听力损失,需要感染效率更高的病毒载体。
发明内容
本公开涉及用于治疗或预防影响受试者内耳的疾病或病症的组合物和方法。发明人已发现包含修饰的AAV衣壳蛋白的重组AAV能够感染内耳毛细胞,从而有效地将遗传物质传递到受试者的内耳毛细胞中。在一些实施方式中,本文提供的组合物和方法可用于治疗或预防受试者的听力损失和/或眩晕。
在一些实施方式中,本公开提供了重组腺相关病毒(AAV)病毒粒子,其包含:(a)修饰的AAV衣壳蛋白,其中该修饰的AAV衣壳蛋白相对于相应的亲本AAV衣壳蛋白包含肽***,其中所述肽***包含氨基酸序列LGETTRP(SEQ ID NO:1),其中该修饰的AAV衣壳蛋白中的***在对应于AAV2的VP1的氨基酸587和588的氨基酸之间;以及(b)产生表达产物的异源核酸,其中该表达产物减少听力损失和/或眩晕。
在一些实施方式中,表达产物是降低与听力损失和/或眩晕相关基因的表达的核酸,其中与听力损失和/或眩晕相关的基因选自下组:DIAPH1、KCNQ4、GJB3、IFNLR1、GJB2、GJB6、MYH1、CEACAM16、GSDME/DFNA5、WFS1、LMX1A、TECTA、COCH、EYA4、MYO7A、COL11A2、POU4F3、MYH9、ACTG1、MYO6、SIX1、SLC17A8、REST、GRHL2、NLRP3、TMC1、COL11A1、CRYM、P2RX2、CCDC50、MIRN96、TJP2、TNC、SMAC/DIABLO、TBC1D24、CD164、OSBPL2、HOMER2、KITLG、MCM2、PTPRQ、DMXL2、MYO3A和PDE1C。
在一些实施方式中,表达产物是减少听力损失和/或眩晕的多肽,其中该多肽选自下组:GJB2、GJB6、MYO7A、MYO15A、SLC26A4、TMIE、TMC1、TMPRSS3、OTOF、CDH23、GIPC3、STRC、USH1C、OTOG、TECTA、OTOA、PCDH15、RDX、GRXCR1、TRIOBP、CLDN14、MYO3A、WHRN、CDC14A、ESRRB、ESPN、MYO6、HGF、ILDR1、ADCY1、CIB2、MARVELD2、BDP1、COL11A2、PDZD7、PJVK、SLC22A4、SLC26A5、LRTOMT/COMT2、DCDC2、LHFPL5、S1PR2、PNPT1、BSND、MSRB3、SYNE4、LOXHD1、TPRN、GPSM2、PTPRQ、OTOGL、TBC1D24、ELMOD3、KARS、SERPINB6、CABP2、NARS2、MET、TSPEAR、TMEM132E、PPIP5K2、GRXCR2、EPS8、CLIC5、FAM65B、DFNB32、EPS8L2、ROR1、WBP2、ESRP1、MPZL2、PRPS1、POU3F4、SMPX、AIFM1和COL4A。
在一些实施方式中,重组AAV病毒粒子选自下组:AAV2、AAV5、AAV8和AAV9。在一些实施方式中,重组AAV病毒粒子是包含经修饰的AAV2-VP1衣壳蛋白的AA2病毒粒子,例如AAV2.7m8病毒粒子。
在一些实施方式中,表达产物是减少与听力损失相关基因的表达的核酸,是干扰RNA。在一些实施方式中,干扰RNA是反义分子、短干扰RNA或miRNA。
在一些实施方式中,AAV病毒粒子产生如下的表达产物:该表达产物减少年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失或外伤所致的听力损失。
在另一实施方式中,本公开提供了用于治疗或预防受试者的内耳毛细胞损伤的方法,包括向具有或有风险发生内耳毛细胞损伤的受试者施用有效量的本文所述的任何重组AAV病毒粒子。
在一些实施方式中,受试者具有或有风险发生年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失和外伤所致的听力损失。
在一些实施方式中,重组AAV病毒粒子感染耳蜗的内毛细胞和外毛细胞。在一些实施方式中,重组AAV病毒粒子感染耳蜗中的胶质样支持细胞。在一些实施方式中,支持细胞为内柱细胞或内指细胞。
在一些实施方式中,重组AAV病毒粒子增加内耳毛细胞再生,例如,耳蜗毛细胞再生。
在另一实施方式中,本公开还提供了用于治疗或预防受试者的听力损失和/或眩晕的方法,包括向具有或有风险发生听力损失和/或眩晕的受试者施用有效量的本文所述的任何重组AAV病毒粒子。
在一些实施方式中,具有或有风险发生听力损失的受试者是如下的受试者:该受试者具有或有风险发生年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失和外伤所致的听力损失。
在一些实施方式中,重组AAV病毒粒子感染受试者的内耳毛细胞,该受试者具有或有风险发生听力损失。在一些实施方式中,内耳毛细胞是耳蜗的内毛细胞和/或外毛细胞。在一些实施方式中,重组AAV病毒粒子感染受试者的耳蜗中的胶质样支持细胞。在一些实施方式中,支持细胞为内柱细胞/或内指细胞。在一些实施方式中,重组AAV病毒粒子增加内耳毛细胞再生,例如,耳蜗毛细胞再生。
在本文提供的任何方法中,重组AAV病毒粒子能够经静脉内施用,经鞘内施用,经鼓膜内(intratypmanically)施用,经圆窗施用,经半规管递送施用,或经镫骨足板造孔术(stapedotomy)施用。在一些实施方式中,重组AAV病毒粒子经由经管造口术施用到受试者的后半规管中。
附图说明
本发明包括以下附图。附图旨在说明组合物和方法的某些实施方式和/或特征,并且补充组合物和方法的任何描述。除非书面描述中明确表示情况如此,否则附图不会限制组合物和方法的范围。
图1a-1h显示,AAV2.7m8高效地感染耳蜗内毛细胞和外毛细胞。(a-f)当将AAV2.7m8-GFP(a)经由后半规管方法注射到新生小鼠内耳中时,在整个耳蜗中高效地感染了IHCs和OHCs。AAV8BP2-GFP(b)注射引起一些IHCs损失(白色箭头)。AAV-DJ-GFP(c)以极低水平感染耳蜗毛细胞。AAV2-GFP(d)、AAV8-GFP(e)和Anc80L65-GFP(f)以高水平感染IHCs,但OHC感染效率低于AAV2.7m8-GFP。GFP表达以绿色显示,Myo7a表达(毛细胞标记物)以红色显示。显示了耳蜗顶部的40倍图像。比例尺为20μm。(g&h)。IHC(g)和OHC(h)感染效率的定量。误差线表示标准误差。关于AAV2.7m8的统计学显著性在误差线上方显示(*表示p<0.05,**表示p<0.01,***表示p<.001)。IHC:内毛细胞。OHC:外毛细胞。
图2a-2b显示,AAV2.7m8在整个耳蜗中感染内毛细胞和外毛细胞。其经由后半规管方法进行AAV2.7m8-GFP注射的小鼠耳蜗的10倍(a)和40倍(b)图像。在整个耳蜗的IHCs和OHCs中均可见GFP表达。GFP表达以绿色示出,Myo7a表达(用于毛细胞的标志物)以红色示出。
图3a-3g显示,AAV2.7m8以较低效率感染前庭毛细胞。(a-f)胞囊的10倍和40倍图像,显示了响应于后管(posterior canal)AAV递送的毛细胞感染效率。AAV2.7m8-GFP(a)、AAV8BP2-GFP(b)、AAV-DJ-GFP(c)、AAV2-GFP(d)以较低水平感染前庭毛细胞。相比之下,AAV8-GFP(e)和Anc80L65-GFP(f)以较高效率感染前庭毛细胞。GFP表达以绿色显示,Myo7a表达(毛细胞标记物)以红色显示。(g)前庭毛细胞感染效率的定量。关于AAV2.7m8的统计学显著性在误差线上方显示(*表示p<0.05,**表示p<0.01,***表示p<.001)。误差线表示标准误差。SSC:上半规管。HSC:水平半规管。
图4a-4g显示,AAV2.7m8高效地感染内柱细胞和内指细胞。(a-f):耳蜗顶部的共焦图像,显示了响应于后管AAV递送的内柱细胞(IPC)和内指细胞(IPhC)感染效率。AAV2.7m8-GFP(a)以高水平感染IPCs和IPhCs。相比之下,AAV8BP2(b)不感染IPCs和IPhCs。AAV-DJ-GFP(c)、AAV2-GFP(d)、AAV8-GFP(e)和Anc80L65-GFP(f)以较低水平感染IPCs,但不感染IPhCs。(g)IPC感染效率的定量。关于AAV2.7m8的统计学显著性在误差线上方显示(*表示p<0.05,**表示p<0.01,***表示p<.001)。误差线表示标准误差。
图5a-5g显示,AAV2.7m8高效地感染内柱细胞和内指细胞。(a-e)经由后半规管方法进行AAV2.7m8-GFP注射的小鼠耳蜗顶部的代表性全图像。内柱细胞和内指细胞显示了高水平的GFP表达。GFP表达以绿色显示,Myo7a表达(毛细胞标记物)以红色显示,乙酰化微管蛋白表达以洋红色显示(支持细胞标记物),Hoechst染色以蓝色显示(核标记物)。显示了40倍图像。(f)显示具有强表达的内柱细胞和内指细胞的同一图像的正交投影。显示了耳蜗顶部的40倍图像。(g和h)内柱细胞(g)和内指细胞(h)感染效率的定量。误差线表示标准误差。IHC:内毛细胞。OHC:外毛细胞。IPC:内柱细胞。IPhC:内指细胞。
图6a-6b显示,AAV2.7m8对接受注射的小鼠的听觉功能和前庭功能有最小的不良作用。(a)记录听性脑干反应(ABR)以评估经由后半规管方法进行合成AAV注射的小鼠的听觉功能。AAV2.7m8、AAV-DJ、AAV2、AAV8和Anc80L65对于听觉功能有最小的不良作用,而与未注射的对照小鼠相比,注射AAV8BP2引起10-25dB的ABR阈值保持升高。(b)评价经由后半规管方法进行AAV注射的小鼠的转圈行为。与未接受注射的对照小鼠相比,AAV2.7m8、AAV-DJ、AAV2、AAV8和Anc80L65未引起统计学显著的转圈行为增加,而与未接受注射的对照小鼠相比,注射AAV8BP2引起转圈行为稍微升高。关于AAV2.7m8的统计学显著性在误差线上方显示(*表示p<0.05,**表示p<0.01,***表示p<.001)。误差线表示标准误差。
图7a-7b显示,AAV8BP2引起耳蜗炎症。(a)在AAV2.7m8-GFP注射之后使用苏木精-曙红(H&e)染色进行耳蜗检查,未见炎性细胞浸润的证据。(b)相比之下,在AAV8BP2注射之后,耳蜗中可见炎性细胞浸润。SV:前庭阶。SM:蜗管。ST:鼓阶。
图8a-8c显示,较低浓度的AAV8BP2未引起听力损失和转圈增加。(a)与未接受注射的对照小鼠相比,以0.5x1010G.C.后管注射AAV8BP2(AAV8BP2 0.5e10)未引起ABR阈值升高。(b)与未接受注射的对照小鼠相比,以0.5x1010 G.C.后管注射AAV8BP2(AAV8BP2 0.5e10)未引起转圈行为升高。(c)与1x1010 G.C.相比,当以0.5x1010 G.C.递送AAV8BP2时,IHCs和OHCs的感染效率较低。从耳蜗顶获取图像。
具体实施方式
以下说明书阐述了本申请组合物和方法的各个方面和实施方式。没有特定的实施方式旨在限定组合物和方法的范围。相反,实施方式仅提供至少被包含在所公开的组合物和方法范围内的各种组合物和方法的非限制性示例。说明书应从本领域普通技术人员的角度解读;因此,不必包括本领域技术人员熟知的信息。
发明人已发现包含修饰的AAV衣壳蛋白的重组AAV能够用于感染受试者的内耳毛细胞并且有效地将遗传物质传递到受试者的内耳毛细胞中。本文提供了用于治疗或预防内耳毛细胞损伤的组合物和方法。在一些实施方式中,本文中提供的组合物和方法可用于治疗或预防影响受试者内耳的疾病或病症。
在一些实施方式中,重组AAV病毒粒子包含a)修饰的AAV衣壳蛋白,其中所述修饰的AAV衣壳蛋白相对于相应的亲本AAV衣壳蛋白包含肽***,其中所述肽***包含氨基酸序列LGETTRP(SEQ ID NO:1),其中所述修饰的AAV衣壳蛋白中的***在对应于AAV2的VP1的氨基酸587和588的氨基酸之间。
在一些实施方式中,重组AAV病毒粒子包含a)修饰的AAV衣壳蛋白,其中所述修饰的AAV衣壳蛋白相对于相应的亲本AAV衣壳蛋白包含肽***,其中所述肽***包含氨基酸序列LGETTRP(SEQ ID NO:1),其中该修饰的AAV衣壳蛋白中的***在对应于AAV2的VP1的氨基酸587和588的氨基酸之间;以及(b)产生表达产物的异源核酸序列,其中该表达产物减少听力损失和/或眩晕。
***可以在AAV2的氨基酸587和588之间,或另一种AAV血清型的衣壳亚单位的对应位置。本领域技术人员能容易地将AAV2-VP1的氨基酸序列与另一种AAV血清型的VP1氨基酸序列的氨基酸序列进行比对,以识别另一种AAV血清型的VP1(例如来自AAV1,AAV3,AAV4,AAV5,AAV6,AAV7,AAV8,AAV9或AAV10的VP1)中对应于AAV2-VP1的氨基酸587和588的氨基酸。***也可以在对应于AAV2-VP1的第570-611个氨基酸或另一种AAV血清型的衣壳亚单位的相应位置的氨基酸中的两个相邻氨基酸之间进行。
包含LGETTRP(SEQ ID NO:1)的多肽***的长度可以为7-15个氨基酸。例如,***的长度可以为7-10个,长度为7-111个氨基酸,长度为7-12个氨基酸,长度为7-13个氨基酸,长度为7-14个氨基酸或长度为7-15个氨基酸。***也可以为约7,8,9,10,11,12,13,14或15个氨基酸。AAV2Vp1衣壳蛋白的氨基酸序列可见于GenBank Accession No.YP_680426.1(SEQ ID NO:2)。在一些实施方式中,***可在对应于氨基酸序列的第570-611个氨基酸中的两个相邻氨基酸之间(例如,在氨基酸587和588之间)进行,所述氨基酸序列与AAV2-VP1的氨基酸序列具有至少75%,80%,90%,95%或99%同一性。
适合用于确定序列同一性和序列相似性百分比的算法是BLAST和BLAST 2.0算法,它们分别在以下中描述:Altschul等人,(1990)J.Mol.Biol.215:403-410和Altschul等人,(1977)Nucleic Acids Res.25:3389-3402。用于执行BLAST分析的软件可通过国家生物技术信息中心(the National Center for Biotechnology Information,NCBI)网站公开获取。该算法首先通过识别查询序列中长度为W的短词来识别高分序列对(HSP),当与数据库序列中长度相同的词比对时,这些短词匹配或满足某个正值阈值T。T被称为邻居字串得分阈值(neighborhood word score threshold)(Altschul等人,同上)。这些最初的邻居字串命中(neightborhood word hits)充当种子以启动搜索,从而找到包含它们的较长HSPs。然后,沿每个序列的两个方向扩展字串命中,直到累积比对得分增加。对于核苷酸序列,使用参数M(匹配残基对的奖励分数;始终>0)和N(不匹配残基的惩罚分数;始终<0)计算累积得分。对于氨基酸序列,使用评分矩阵来计算累积得分。当:累计比对得分降低到低于其最大实现值减去数量X时,停止向每个方向扩展字串命中数;由于一个或多个负得分残基比对的累积,累积得分归零或更低;或者到达任一序列的末尾。BLAST算法的参数W、T和X决定了比对的灵敏度和速度。BLASTN程序(用于核苷酸序列)使用以下作为默认值:字长(W)为28,期望值(E)为10,M=1,N=-2,并且比较双链。对于氨基酸序列,BLASTP程序使用以下作为默认值:字长(W)为3,期望值(E)为10,BLOSUM62评分矩阵(参见Henikoff&Henikoff,Proc.Natl.Acad.Sci.USA 89:10915(1989))。
BLAST算法还执行两个序列之间的相似性的统计学分析(参见,例如,Karlin&Altschul,Proc.Nat'l.Acad.Sci.USA 90:5873-5787(1993))。BLAST算法提供的一种相似性量度为最小和概率(P(N)),其提供了两个核苷酸序列或氨基酸序列之间偶然匹配的概率指示。例如,如果在测试核酸与参比核酸的比较中最小和概率小于约0.01,更优选小于约10-5,最优选小于约10-20,则认为该核酸与参比序列相似。
在一些实施方式中,重组AAV病毒粒子是AAV2.7m8病毒粒子。参见,例如,Dalkara,D.等人,(In vivo-directed evolution of a new adeno-associated virus fortherapeutic outer retinal gene delivery from the vitreous.Sci Transl Med5,189ra176(2013))和美国专利第9,193,956号,以上文献通过引用以其整体在此并入。在一些实施方式中,与AAV VP1衣壳蛋白中所含的没有肽***的相应亲本AAV衣壳蛋白的重组AAV病毒粒子的内耳毛细胞感染性相比,重组AAV病毒粒子提供了增加的内耳毛细胞(例如耳蜗毛细胞)传染性。在一些实施方式中,与重组AAV8BP2病毒粒子或重组AAV Anc80L65病毒粒子的内耳毛细胞感染性相比,重组AAV病毒粒子(例如AAV2.7m8)提供了增加的内耳毛细胞(例如耳蜗毛细胞)感染性。在施用本文所述的重组AAV病毒粒子(例如AAV2.7m8)之后,与对照相比,增加的内耳毛细胞感染性可以为约5%、10%、20%、30%、40%、50%、60%、70%、80%、90%、100%或更多增加。该增加也可以是至少2倍、5倍、10倍、20倍、30倍、40倍、50倍或更多倍增加。
如本文所用,重组AAV病毒粒子是包含至少一种AAV衣壳蛋白和包封的重组AAV载体的病毒颗粒。如本文所用,"重组AAV载体"是指包含通常不存在于AAV中的核酸序列(即,与AAV异源的多核苷酸)的AAV载体,例如,用于细胞遗传转化的核酸序列。一般来说,异源核酸的两侧至少有一个,而且通常有两个AAV反向末端重复序列(ITRs)。术语重组AAV载体包括rAAV载体颗粒和重组AAV载体质粒。重组AAV载体可以是单链的(ssAAV)或自互补的(scAAV)。
AAV的各种血清型的基因组序列以及天然末端重复序列(TRs)、Rep蛋白和衣壳亚单位的序列在本领域中是已知的。此类序列可以在文献中或在例如GenBank等公共数据库中找到。参见,例如,GenBank Accession Numbers NC_002077(AAV-1),AF063497(AAV-1),NC_001401(AAV-2),AF043303(AAV-2),NC_001729(AAV-3),NC_001829(AAV-4),U89790(AAV-4),NC_006152(AAV-5),AF513851(AAV-7),AF513852(AAV-8),和NC_006261(AAV-8);其公开内容通过引用并入本文用于教导AAV核酸和氨基酸序列。
“AAV病毒”、“AAV病毒粒子”、“AAV病毒颗粒”或“重组AAV载体颗粒”是指由至少一种AAV衣壳蛋白和包封的多核苷酸重组AAV载体组成的病毒颗粒。如果颗粒包含异源核酸序列(即不同于野生型AAV基因组的核酸序列,例如将被递送到哺乳动物细胞的转基因),则其可被称为重组AAV载体。因此,重组AAV颗粒或病毒粒子的产生必然包括重组AAV载体的产生,因此这样的载体包含在重组AAV颗粒内。用于产生AAV载体和病毒粒子的方法是本领域已知的。参见,例如,Shin等人,“Recombinant Adeno-Associated Viral VectorProduction and Purification,”Methods Mol.Biol.798:267-284(2012))。本文所述的任何AAV病毒粒子可用于感染一种或多种类型的内耳毛细胞,包括但不限于耳蜗细胞、前庭细胞、耳蜗内毛细胞、耳蜗外毛细胞、耳蜗的胶质样支持细胞(例如,Hensen细胞、Deiters细胞、内外柱细胞、Claudius细胞和内指细胞)。
如通篇所用,“相应的亲本AAV衣壳蛋白”是指没有肽***的相同AAV血清型的AAV衣壳蛋白。如本文所用,当描述重组AAV载体或病毒粒子时,短语“异源”指在野生型AAV中未天然发现的核酸序列。例如,产生表达产物的异源核酸序列是通常不能在野生型AAV中找到的核酸。在异源核酸序列编码多肽的实施方式中,编码的多肽是为通常不是由天然的野生型AAV编码或表达的异源多肽。
如通篇所用,“表达产物”是在被AAV病毒粒子感染后在细胞(例如内耳毛细胞)中表达或产生的核酸序列或多肽。表达产物可通过体外、体内或体外感染细胞表达。如在本说明书和所附权利要求书中所使用的,除非上下文另有明确规定,否则单数形式“一(a)”、“一(an)”和“所述/该(the)”包括复数指代物。因此,术语“病毒粒子”或“细胞”也指多于一个病毒粒子或细胞,例如,病毒粒子或细胞群。
表达产物包括但不限于多肽、适配子、反义分子、干扰RNA或mRNA。在一些实施方式中,表达产物是选自下组的干扰RNA:短干扰RNA(siRNA)、短发夹(shRNA)和miRNA。
如通篇所用,术语“核酸”是指脱氧核糖核酸(DNA)或核糖核酸(RNA)及其单链或双链形式的聚合物。除非特别限定,否则该术语包括含有天然核苷酸的已知类似物的核酸,所述核酸具有与参考核酸相似的结合特性并且以类似于天然核苷酸的方式代谢。除非另有说明,特定核酸序列还隐含地包含其保守修饰变体(例如,简并密码子替换)、等位基因、同源序列、SNPs和互补序列以及明确指出的序列。具体地说,简并密码子替换可通过生成如下序列来实现:其中一个或多个选定的(或全部的)密码子的第三位置被混合碱基和/或脱氧核糖核酸残基替换(Batzer等人,Nucleic Acid Res.19:5081(1991);Ohtsuka等人,J.Biol.Chem.260:2605-2608(1985);和Rossolini等人,Mol.Cell.Probes 8:91-98(1994))。
在一些实施方式中,编码目标表达产物的核酸序列可操作地连接到组成型启动子。在其它实施方式中,编码目标表达产物的核酸序列可操作地连接到诱导型启动子。在一些情况中,编码目标表达产物的核酸序列可操作地连接到组织特异性或细胞类型特异性调控元件。例如,在一些情况中,编码目标表达产物的核酸序列可操作地连接到内耳毛细胞特异性调控元件,例如,使可操作地连接的核酸在内耳毛细胞中选择性表达的调控元件。关于基因产物在MY07A启动子控制下在内耳细胞中的表达,参见,例如,Boeda and Petit“Aspecific promoter of the sensory cells of the inner ear defined bytransgenesis”Hum Mol.Genet.19(15):1581-9(2001)。如本文所用,特异性表达并不意味着表达产物仅在特定组织或细胞类型中表达,而是指基本上限于特定组织或细胞类型的表达。产生表达产物的任何异源核酸可进一步包含编码可检测多肽(例如荧光多肽(GFP、RFP等)或其活性片段)的核酸。
当用本文所述的任何AAV病毒粒子感染受试者的内耳毛细胞时,在内耳毛细胞中产生的表达产物减少了受试者的听力损失和/或眩晕。在一些实施方式中,表达产物是降低与受试者听力损失和/或眩晕相关基因的表达的核酸序列,例如,反义分子或干扰RNA。
在一些实施方式中,核酸序列(例如,反义分子或干扰RNA)降低选自下组的一个或多个基因在受试者内耳毛细胞中的表达:DIAPH1,KCNQ4,GJB3,IFNLR1,GJB2,GJB6,MYH1,CEACAM16,GSDME/DFNA5,WFS1,LMX1A,TECTA,COCH,EYA4,MYO7A,COL11A2,POU4F3,MYH9,ACTG1,MYO6,SIX1,SLC17A8,REST,GRHL2,NLRP3,TMC1,COL11A1,CRYM,P2RX2,CCDC50,MIRN96,TJP2,TNC,SMAC/DIABLO,TBC1D24,CD164,OSBPL2,HOMER2,KITLG,MCM2,PTPRQ,DMXL2,MYO3A和PDE1C。在一些实施方式中,表达减少是mRNA的转录减少和/或从mRNA翻译的多肽或其片段的翻译减少。与对照相比,表达的减少或降低可以是约5%、10%、20%、30%、40%、50%、60%、70%、80%、90%、100%或这些百分比之间的任何百分比的减少或降低。通过降低选自下组的一个或多个基因的表达:DIAPH1,KCNQ4,GJB3,IFNLR1,GJB2,GJB6,MYH1,CEACAM16,GSDME/DFNA5,WFS1,LMX1A,TECTA,COCH,EYA4,MYO7A,COL11A2,POU4F3,MYH9,ACTG1,MYO6,SIX1,SLC17A8,REST,GRHL2,NLRP3,TMC1,COL11A1,CRYM,P2RX2,CCDC50,MIRN96,TJP2,TNC,SMAC/DIABLO,TBC1D24,CD164,OSBPL2,HOMER2,KITLG,MCM2,PTPRQ,DMXL2,MYO3A和PDE1C,可以降低或改善听力损失。
在一些实施方式中,表达产物是降低或改善受试者的听力损失和/或眩晕的多肽。如通篇所用,“多肽”、“肽”和“蛋白质”在本文中可互换地用于指代氨基酸残基的聚合物。如本文所用,该术语包含任何长度的氨基酸链,包括全长蛋白质,其中氨基酸残基通过共价肽键连接。本文所述的任何多肽的片段也被这些术语所涵盖。
在一些实施方式中,在受试者的内耳毛细胞表达选自下组的一种或多种多肽:GJB2,GJB6,MYO7A,MYO15A,SLC26A4,TMIE,TMC1,TMPRSS3,OTOF,CDH23,GIPC3,STRC,USH1C,OTOG,TECTA,OTOA,PCDH15,RDX,GRXCR1,TRIOBP,CLDN14,MYO3A,WHRN,CDC14A,ESRRB,ESPN,MYO6,HGF,ILDR1,ADCY1,CIB2,MARVELD2,BDP1,COL11A2,PDZD7,PJVK,SLC22A4,SLC26A5,LRTOMT/COMT2,DCDC2,LHFPL5,S1PR2,PNPT1,BSND,MSRB3,SYNE4,LOXHD1,TPRN,GPSM2,PTPRQ,OTOGL,TBC1D24,ELMOD3,KARS,SERPINB6,CABP2,NARS2,MET,TSPEAR,TMEM132E,PPIP5K2,GRXCR2,EPS8,CLIC5,FAM65B,DFNB32,EPS8L2,ROR1,WBP2,ESRP1,MPZL2,PRPS1,POU3F4,SMPX,AIFM1和COL4A或其片段。
在一些实施方式中,当用本文所述的重组AAV病毒粒子感染受试者的内耳毛细胞时,与对照相比,受试者的内耳毛细胞中的一种或多种多肽的水平有至少2倍,至少5倍,至少10倍,至少20倍,至少30倍,至少40倍,至少50倍或至少50倍以上的增加,从而降低了受试者的听力损失和/或眩晕。
表达产物可以与受试者的细胞异源。如本文所用,短语“异源”在涉及细胞(例如,受试者的内耳毛细胞)中的表达产物时是指不能在受试者的细胞中天然发现的核酸序列或多肽。术语"异源序列"是指在指定细胞中本质上不能正常发现的序列。因此,异源核苷酸或蛋白质序列可以是:(a)对其宿主细胞是外来的(即,对该细胞是外源的);(b)在宿主细胞中自然存在(即内源性),但在细胞中以非自然数量存在(即比在宿主细胞中自然存在的数量多或少);或(c)在宿主细胞中自然发现,但位于其自然位置之外。
方法
本文提供了用于通过施用本文所述的任何AAV病毒粒子来将目标核酸递送到内耳的方法。在一些实施方式中,AAV病毒粒子包含目标核酸。在一些实施方式中,目标核酸被递送到内耳毛细胞,例如,耳蜗细胞。在一些实施方式中,AAV病毒粒子是包含目标核酸的AAV2.7m8病毒粒子。在一些实施方式中,目标核酸减少内毛细胞损伤,降低听力损失和/或降低眩晕。在一些实施方式中,目标核酸编码减少内毛细胞损伤,降低听力损失和/或降低眩晕的多肽。
听力损失通常因内耳毛细胞(例如耳蜗毛细胞)受损引起。哺乳动物耳蜗包含两种类型的毛细胞,内毛细胞(IHCs)和外毛细胞(OHCs),二者对于听觉信息的检测和处理都有重要意义。这些毛细胞被支持细胞所围绕,所述支持细胞是一组异质细胞,其对于耳蜗内稳态有重要意义。成熟的哺乳动物毛细胞不能再生。因此,一旦这些细胞发生损伤,退化过程通常是不可逆的。
本文提供了治疗或预防受试者的内耳毛细胞损伤的方法,包括对具有内耳毛细胞损伤或有风险发生内耳毛细胞损伤的受试者施用有效量的本文所述的重组AAV病毒粒子。在一些实施方式中,重组病毒粒子是包含减少与内耳毛细胞损伤相关基因的表达的核酸序列的重组AAV病毒粒子,例如,AAV2.7m8病毒粒子。在一些实施方式中,重组AAV病毒粒子是包含编码治疗或预防受试者的内耳毛细胞损伤的多肽的核酸序列的重组AAV2病毒粒子,例如,AAV2.7m8病毒粒子。在一些实施方式中,具有内耳毛细胞损伤或有风险发生内耳毛细胞损伤的受试者具有听力损失或有风险发生听力损失。在一些实施方式中,具有内耳毛细胞损伤或有风险发生内耳毛细胞损伤的受试者经历眩晕。
在另一实施方式中,本文提供了治疗或预防受试者的听力损失和/或眩晕的方法,包括对具有听力损失或眩晕或有风险发生听力损失或眩晕的受试者施用有效量的本文所述的重组AAV病毒粒子。在一些实施方式中,重组AAV病毒粒子是包含减少与内耳毛细胞损伤相关基因的表达的核酸序列的重组AAV2病毒粒子,例如,AAV2.7m8病毒粒子。在一些实施方式中,重组病毒粒子是包含编码治疗或预防受试者的内耳毛细胞损伤的多肽的核酸序列的重组AAV病毒粒子,例如,AAV2.7m8病毒粒子。
在一些实施方式中,重组AAV病毒粒子增加内耳毛细胞再生,例如,耳蜗毛细胞再生。在一些实施方式中,重组AAV病毒粒子感染耳蜗的内毛细胞和/或外毛细胞。在一些实施方式中,重组AAV病毒粒子感染耳蜗中的胶质样支持细胞。在一些实施方式中,重组AAV病毒粒子所感染的支持细胞是内柱细胞和/或内指细胞。在一些实施方式中,重组AAV病毒粒子增加了耳蜗的内毛细胞,外毛细胞和/或胶质样支持细胞的再生。在一些实施方式中,重组AAV病毒粒优先感染耳蜗毛细胞。在一些实施方式中,受试者内耳中耳蜗毛细胞的重组AAV病毒粒子感染效率比受试者内耳中前庭细胞中的重组AAV病毒粒子感染效率高至少5%,10%,20%,30%,40%,50%,60%,70%,80%,90%或至少100%。在一些实施方式中,受试者内耳中重组AAV病毒粒子产生的表达产物在耳蜗细胞中的水平与受试者内耳中前庭细胞相比高至少5%,10%,20%,30%,40%,50%,60%,70%,80%,90%或至少100%。
本文提供的方法和组合物可用于治疗具有或有风险发生任何类型听力损失的受试者。听力损失可以在传导水平、感音神经和/或中枢水平上。传导性听力损失由涉及外耳或中耳的损害引起,导致鼓膜和小骨向内耳液放大的空气传播声音的正常通路被破坏。感音神经性听力损失由耳蜗或八颅神经的听觉分支的损害引起。中枢性听力损失由于中枢听觉通路的损害引起。在某些情况下,传导性听力损失与感音神经性听力损失合并发生(混合性听力损失)。
举例来说,本文中提供的组合物和方法可用于治疗具有或有风险发生年龄相关的听力损失(老年性耳聋),遗传性听力损失,噪音诱导的听力损失,疾病相关的听力损失,暴露于毒性物质和因创伤所致的听力损失的受试者。
在一些实施方式中,遗传性听力损失可由涉及听力的一个或多个基因的突变引起。有些突变会导致非综合征性听力损失,这意味着受试者除了听力损失之外没有任何其他症状。导致听力损失的其他突变是综合征性的,这意味着此人除了听力损失之外还有其他症状(例如,Waardenburg综合征、Alport综合征和Usher综合征)。在一些实施方式中,遗传性听力损失是常染色体显性听力损失,例如,由GJB2突变引起的听力损失。
在一些实施方式中,将编码与听力损失相关的缺失或突变基因的未突变多肽的核酸序列递送到受试者的内耳毛细胞以向内耳毛细胞提供参与听力损失的缺失或突变基因的工作拷贝。在其它实施方式中,将降低参与听力损失的基因的一个或多个突变等位基因的表达的核酸序列递送到受试者的内耳毛细胞。
本文中提供的组合物和方法也可以用于治疗具有或有风险发生眩晕的受试者。在一些实施方式中,眩晕与前庭病症有关。前庭病症的示例包括但不限于良性阵发性位置性眩晕(BPPV),迷路炎,前庭神经炎,Ménière病,继发性内淋巴积液,外淋巴瘘。前庭病症还包括上半规管裂,听神经瘤,耳毒性,大前庭水管综合征,和mal de débarquement。
本文提供的治疗听力损失或眩晕的任何方法可与用于听力损失或眩晕的其他治疗相结合,例如助听器,施用有效量的皮质类固醇,或用于治疗眩晕的练习等。
在通篇中,治疗(treat,treating和treatment)是指减少或延迟听力损失的一种或多种效果或症状(例如,听不懂讲话、听电视或收音机音量过大、耳鸣、要求人们重复自己的话)或眩晕(例如,失去平衡、晕厥、复视、意识模糊、口齿不清、四肢麻木)。受试者可以被诊断患有听力损失或眩晕。治疗也可以指减少潜在病理而不仅仅是症状的方法。对受试者施用的效果可具有以下效果,但不限于这些:减轻疾病的一个或多个症状、减轻疾病的严重性、完全消除疾病,或延迟一个或多个症状的发作或恶化。例如,如果与治疗前的受试者相比或与对照受试者或对照值相比,受试者的听力损失或眩晕有至少约10的减少%,则所公开的方法被认为是治疗。因此,减少可以是约10、20、30、40、50、60、70、80、90、100%或其中的任何减少量。减少听力损失也可以是听力改善了至少约5、10、20、30、40、50、60、70、80、90、100%或这些百分比之间的任何百分比。用于测试受试者听力的方法是本领域已知的,并且包括:
如本文所用,预防(prevent,preventing或prevention)表示排除、延迟、避免、避免、预防、停止或阻碍疾病或病症的发生、发生、严重性或复发的方法。例如,如果听力损失或眩晕或者听力损失的一个或多个症状(例如,听不懂讲话、听电视或收音机音量过大、耳鸣、要求人们重复自己的话)或眩晕的一个或多个症状(例如,失去平衡、晕厥、复视、意识模糊、口齿不清、四肢麻木)的发作、发生、严重性或复发有所减少,则所公开的方法被认为是预防。听力丧失或眩晕的发作、发生、严重性或复发的减少或延迟可为约10、20、30、40、50、60、70、80、90、100%或其间的任何减少量。
如通篇所用,受试者表示个体。受试者可以是成年受试者或未成年受试者。未成年受试者包括年龄在初生到18岁范围内的受试者。因此,不到约10岁,五,两岁,一岁,六个月,三个月,一个月,一周,或一天的未成年受试者也被包含作为受试者。优选地,受试者是哺乳动物,如灵长类动物,更优选人。非人灵长类动物也是受试者。术语受试者包括家养动物(如猫、狗等),家畜(例如,牛、马、猪、绵羊、山羊等)和实验室动物(例如,雪貂、会输、小鼠、兔子、大鼠、沙鼠、豚鼠等)。因此,本文中设想了兽医学用途和医药制剂。
药物组合物
本文提供了药物组合物,其包含本文所述的任何重组AAV病毒粒子和药学上可接受的载剂,稀释剂,赋形剂,或缓冲剂。在一些实施方式中,药学上可接受的载剂,稀释剂,赋形剂,或缓冲剂适合用于受试者(例如人)中。可将药物组合物递送到受试者中,以允许在受试者的内耳细胞中产生表达产物。药物组合物包含允许接受者产生有效量的减少或预防内毛细胞损伤的表达产物的足够遗传物质。在一些实施方式中,药物组合物包含允许接受者产生有效量的治疗或预防受试者的听力损失和/或眩晕的表达产物的足够遗传物质。
组合物可单独施用或与至少一种其它药剂(例如稳定化合物)组合施用,其可在任何无菌的、生物相容的药物载剂中施用,包括但不限于盐水、缓冲盐水、葡萄糖和水。在一些实施方式中,药物组合物还包含药学上可接受的赋形剂。此类赋形剂包括本身不会诱导对接受组合物的个体有害的免疫应答并且可以在没有过度毒性的情况下施用的任何药物剂。药学上可接受的赋形剂包括但不限于液体,例如水、盐水、甘油、糖和乙醇。其中可包含药学上可接受的盐,例如,无机酸盐,例如盐酸盐、氢溴酸盐、磷酸盐、硫酸盐等;以及有机酸盐,例如乙酸盐、丙酸盐、丙二酸盐、苯甲酸盐等。此外,例如润湿剂或乳化剂、pH缓冲物质等的辅助物质可存在于此类溶媒中。药物上可接受的载剂、赋形剂和含有这些材料的制剂的制备在例如以下中描述:Remington:The Science and Practice of Pharmacy,22ndedition,Loyd V.Allen et al,editors,Pharmaceutical Press(2012).
适合于胃肠外施用的药物制剂可在水溶液中配制,优选在生理相容缓冲剂中,例如Hanks溶液、Ringer溶液或生理缓冲盐水。水性注射悬浮液可以含有增加悬浮液粘度的物质,例如羧甲基纤维素钠、山梨醇或葡聚糖。另外,活性化合物的悬浮液可以根据需要制备为油性注射悬浮液。合适的亲脂性溶剂或溶媒包括脂肪油(如芝麻油),或合成脂肪酸酯(如油酸乙酯或甘油三酯),或脂质体。任选地,悬浮液还可包含合适的稳定剂或试剂,其增加化合物的溶解度以允许制备高浓度溶液。
递送方法
本公开提供了将表达产物递送至个体的内耳毛细胞的方法,该方法包括对个体施用如上所述的重组AAV病毒粒子。表达产物可以是例如多肽、反义分子、干扰RNA或适配子。
如通篇所用,术语“有效量”定义为产生所需的生理反应(例如,减少或预防内耳毛细胞损伤)所必需的任何量。用于施用本文所述重组AAV病毒粒子的有效量和时间表可以经验地确定,并且进行此类确定在本领域技术范围内。施用的剂量范围是那些足够大以产生所需效果的范围,在所需效果中,疾病或病症的一个或多个症状受到影响(例如,减少或延迟)。剂量不应该大到以致引起显著不良副作用,例如不想要的交叉反应、不想要的细胞死亡等。通常,剂量将随抑制剂类型,受试者的种类、年龄、体重、一般健康、性别和饮食,施用方式和时间,以及特定状况的严重程度而变化,并且可由本领域技术人员确定。如有任何禁忌症,可由个别医生调整剂量。剂量可以变化,并且可以以一个或多个剂量施用。
本文所述的任何重组AAV病毒粒子的有效量将变化,并且可由本领域技术人员通过实验和/或临床试验来确定。例如,对于体内注射,例如,直接注射到受试者内耳,有效剂量可为约106至约1015个重组rAAV病毒粒子,例如,约108至1012个重组AAV病毒粒子。对于体外感染,递送到细胞的有效量的重组病毒粒子可以为约106至约1015个重组AAV病毒粒子。其他有效剂量可由本领域普通技术人员通过建立剂量-反应曲线的常规试验容易地建立。
本文所述的组合物以多种方式施用,取决于是否需要局部或全身治疗。组合物通过以下几种施用途径中的任何一种施用:静脉内,鞘内,鼓膜内(intratypmanically)施用,经圆窗施用,经半规管递送,或经镫骨足板造孔术(stapedotomy)施用。在一些实施方式中,组合物以经管造口术施用到受试者的后半规管中。本文所述的任何施用方法的有效剂量可从来自体外或动物模型试验***的剂量-反应曲线推断。
公开了可用于、可与之结合使用、可用于制备所公开的方法和组合物或是所公开的方法和组合物的产品的材料、组合物和组分。本文公开了这些和其他材料,并且可以理解,当公开了这些材料的组合、子集、相互作用、基团等时,虽然可以不明确地公开这些化合物的每个不同的单独和集体组合和排列的具体引用,每一个都在本文中具体地考虑和描述。例如,如果公开和讨论了一种方法,并且讨论了可以对包括在该方法中的多个分子进行的多种,除非特别地指示相反,否则该方法的每一个组合和排列以及可能的修改都被特别地考虑。同样,还具体地考虑和公开了这些的任何子集或组合。此概念适用于本公开的所有方面,包括但不限于使用所公开组合物的方法中的步骤。因此,如果有各种可以执行的附加步骤,则可以理解,这些附加步骤中的每一个都可以与所公开方法的任何特定方法步骤或方法步骤的组合一起执行,并且每一个这样的组合或组合的子集都是特别考虑的并且应该被认为是公开的。
本文引用的出版物及其引用的材料通过引用以其整体具体并入。
实施例
提供以下实施例仅作为说明而非限制。本领域技术人员将容易地认识到可以改变或修各种非临界参数改以产生基本相同或类似结果。
方法
AAV载体构建
由Research Vector Core at the Center for Advanced Retinal and OcularTherapeutics(先进视网膜和眼科治疗中心载体研究中心(宾夕法尼亚大学))生产AAV2.7m8-CAG-eGFP、AAV8BP2-CAG-eGFP、AAV2-CAG-eGFP、AAV2/8-CAG-eGFP和Anc80L65-CAG-eGFP。这些病毒的生产方法如以下中所述:Ramachandran等人,(Evaluation of Doseand Safety of AAV7m8 and AAV8BP2 in the Non-Human Primate Retina.Hum GeneTher 28,154-167(2017))。AAV-DJ-CAG-eGFP购自Vector Biolabs(Malvern,PA)。每种病毒的病毒母液浓度为每毫升1x1013基因组拷贝(G.C.)。
动物外科手术
动物外科手术由Animal Care and Use Committee at National Institute onDeafness and Other Communication Disorders(国家耳聋和其他沟通障碍研究所的动物护理和使用委员会)批准(NIDCD ASP1378-18)。采用低温诱导和维持新生小鼠麻醉(P0-P5)。外科手术只在每只动物的左耳进行。右耳作为对照。在以下文献中描述了通过后半规管方法的内耳基因递送(Isgrig,K.等人,Gene Therapy Restores Balance and AuditoryFunctions in a Mouse Model of Usher Syndrome.Mol Ther 25,780-791(2017))。简单地说,制造耳后切口,解剖组织以暴露后半规管。解剖过程中注意避开面神经。纳升微注射***(Nanoliter2000,World Precision Instruments,Sarasota,FL)与玻璃微移液管配合用于将AAV-GFP加载到玻璃微移液管中。经过约40秒注射总共1μlAAV-eGFP。用5-0号vicryl缝线闭合切口。
听性脑干反应
使用听性脑干反应(ABR)来评价~P30的听敏度。经由腹腔注射***(100mg/kg)和甲苯噻嗪(10mg/kg)麻醉动物,并将其放置在音响室(ETS-Lindgren Acoustic Systems,Cedar Park,TX)内的暖垫上。动物体温使用闭合反馈回路维持并且使用直肠探针监控(CWEIncorporated,TC-1000,Ardmore,PN)。将皮下针状电极***顶点(+)和测试耳乳突(-),接地电极置于对侧耳下。使用Tucker Davis Technologies硬件(RZ6 Multi I/O Processor,Tucker-Davis Technologies,Gainesville,FL,USA)和软件(BioSigRx,v.5.1)完成刺激生成和ABR记录。在如下条件下测量ABR阈值:在4、8、16和32khz下,使用以29.9/秒长线的、具有交替刺激极性的3ms、Blackman门控短音。在每个刺激水平,对512-1024次反应进行平均。通过对波形的目视检查来确定阈值,将其定义为能够可靠地检测到任何波的最低刺激水平。在阈值水平上获得最少两个波形,以确保反应的可重复性。对单个频率的生理学结果进行分析,然后从4到32khz对每个频率取平均值。
转圈行为
使用光学跟踪和ANY-maze跟踪软件(4.96版本,Stoelting Co.,Wood Dale,IL)来定量经历内耳基因递送的小鼠的转圈行为。将38cm x 58cm的箱子附接到摄影机上(Fujinon YV5X2.7R4B-2 1/3-inch 2.7-13.5mm F1.3Day/Night Aspherical Vari-FocalLens)。ANY-maze图像跟踪软件设为跟踪放置在箱子内的小鼠头部。将每只小鼠放置到箱子中,并且使其在新环境中适应2分钟。在接下来的2分钟记录和定量完整的旋转,然后是1分钟“冷静期”,期间不跟踪旋。每只小鼠评估三次,取平均值。
免疫组化和定量
在功能测试完成后,用CO2窒息法使小鼠安乐死,随后断头。取颞骨,用4%多聚甲醛固定过夜,然后在120mM EDTA中脱钙4天。对前庭器官和耳蜗感觉上皮进行显微解剖,阻断,并且用以下物质标记:用于标记毛细胞的小鼠抗肌球蛋白7a抗体(1:200,ProteusBioSciences,Ramona,CA),用于标记支持细胞的小鼠抗乙酰化微管蛋白抗体(1:100,Sigma-Aldrich Corp.,St.Louis,MO),鸡抗GFP抗体(1:1000,Abcam,Cambridge,MA),和标记核的Hoechst染色(1:500,Life Technologies,Carlsbad,CA)。一抗和二抗在PBS中稀释。使用蔡司LSM780共焦显微镜在10倍和40倍的条件下使用z-stacks获得图像。
对于苏木精和曙红(H&E)染色,首先用蔗糖梯度(在PBS中10-30%)处理组织,然后用蔗糖和包埋培养基SCEM(日本Section-Lab Co Ltd)的混合物处理。在液氮中冷冻后,随后将组织在10℃切片为10μm厚度,并且根据制造商的说明使用苏木精-曙红染色试剂盒(Vector Laboratories,Inc.,Burlingame,CA USA)H&E染色。
为定量检测耳蜗毛细胞及支持细胞感染效率,分别在耳蜗顶部、中部、底部各取两张40倍图像。在沿耳蜗的每个位置处计算具有GFP表达的毛细胞和支持细胞数并取平均值。每个40倍图像包含~30个IHC和~90个OHC。通过对整个耳蜗的感染效率进行平均来计算总感染效率。为定量胞囊毛细胞感染效率,对每个胞囊标本采集两张40倍图像(每个图像包含~300个前庭毛细胞),计数并平均具有GFP表达的毛细胞数。
统计
使用Student’s t检验评估感染效率的差异。使用方差分析(ANOVA)评估ABR阈值和转圈行为的差异。采用Scheffe方法进行事后分析。p值<0.05表明统计学显著性。
结果
许多种形式的遗传性听力损失都有影响耳蜗毛细胞的突变,所述耳蜗毛细胞是允许声音检测和处理的机械感觉细胞。研究了3种合成AAVs(AAV2.7m8、AAV8BP2、AAV-DJ)在小鼠内耳的感染模式。AAV2.7m8高效地感染IHCs和OHCs。此外,AAV2.7m8高效地感染内柱细胞和内指细胞。这些结果表明,AAV2.7m8是用于内耳基因疗法的极好的病毒载体。AAV2.7m8极大地拓展了内耳基因疗法的应用领域。
AAV2.7m8使用如下的体内定向进化方法产生:其中通过玻璃体内注射筛选具有不同衣壳蛋白修饰的AAV文库对小鼠光感受器细胞的感染效率(Dalkara等人,)。该载体包含在AAV2衣壳蛋白序列第588位处***的10氨基酸肽,其与AAV2与其主要受体硫酸乙酰肝素蛋白多糖的结合有关(Dalkara et al.;and Khabou et al.,Insight into themechanisms of enhanced retinal transduction by the engineered AAV2 capsidvariant-7m8.Biotechnol Bioeng 113,2712-2724(2016))。类似地,AAV8BP2使用如下的体内定向进化方法产生:其中通过视网膜下注射筛选AAV文库对小鼠视网膜双极细胞的感染。该载体包含在AAV8衣壳蛋白序列第585-594个氨基酸处的修饰(Cronin,T.等人,Efficienttransduction and optogenetic stimulation of retinal bipolar cells by asynthetic adeno-associated virus capsid and promoter.EMBO Mol Med 6,1175-1190(2014))。除AAV2.7m8和AAV8BP2外,另一种已用于各种器官***的合成AAV是AAV-DJ19。AAV-DJ使用如下的DNA家族重组技术产生:其中病毒衣壳含有各种AAV血清型(AAV2、4、5、8和9)的成分(Grimm,D.等人,In vitro and in vivo gene therapy vector evolutionvia multispecies interbreeding and retargeting of adeno-associatedviruses.Journal of virology 82,5887-5911(2008))。AAV-DJ已被证明能感染肝细胞、角质形成细胞、神经元和味觉细胞。
为评价合成AAVs对哺乳动物内耳的感染效率,采用后半规管方法将AAV2.7m8-GFP、AAV8BP2-GFP和AAV-DJ-GFP递送到新生(P0-P5)小鼠内耳。后半规管基因递送允许病毒载体有效地感染耳蜗和前庭器官中的细胞(Isgrig,K.等人,Gene Therapy RestoresBalance and Auditory Functions in aMouse Model of Usher Syndrome.Mol Ther 25,780-791(2017);Tao,Y.等人,Delivery of Adeno-Associated Virus Vectors in AdultMammalian Inner-Ear Cell Subtypes Without Auditory Dysfunction.HumGene Ther(2018);and Suzuki et al.Cochlear gene therapy with ancestral AAV in adultmice:complete transduction of inner hair cells without cochleardysfunction.Scientific reports 7,45524(2017))。还使用相同的递送方法检验了AAV2-GFP和AAV8-GFP(两种常用的常规AAVs,AAV2.7m8和AAV8BP2分别来源于这两种AAVs)以及合成AAV Anc80L65 GFP的感染效率作为额外的对照。将约1x1010基因组拷贝(G.C.)递送到每只动物的内耳。通过用绿色荧光蛋白(GFP)定量毛细胞百分率(用抗Myo7a抗体鉴定)来评价毛细胞感染效率。在基因递送后4周检查耳蜗,显示接受AAV2.7m8-GFP注射的小鼠的IHCs和OHCs二者中的GFP水平均较高(n=8,图1,表1)。IHC的总感染效率为84.1%±5.66%(平均值±标准误差),OHC的总感染效率为83.1±6.17%。接受AAV8BP2-GFP(n=9,图1a-1h,表1)注射的小鼠的IHCs和OHCs中具有中到高水平的GFP表达。IHC的总感染效率为55.7%±为9.53%,并且OHC的总感染效率为44.0%±7.91%(当与AAV2.7m8相比时,IHC和OHC分别为p=0.016和<0.001)。相比之下,接受AAV-DJ-GFP注射的小鼠(n=5,图1,表1)在IHCs和OHCs中仅由低水平的GFP表达。IHC的总感染效率为1.63±1.27%,并且OHC的总感染效率为0.05±0.05当与AAV2.7m8相比时,IHC和OHC均为p<.001)。
表1
AAVs在内耳不同细胞类型中的感染效率。显示了感染效率(%)和标准误差(括号内)显示。IHC:内毛细胞。OHC:外毛细胞。IPC:内柱细胞。IPhC:内指细胞。
AAV2.7m8-GFP与传统AAVs的比较也显示出更高的耳蜗毛细胞感染效率,特别是对于OHCs。对于AAV2-GFP(n=3,图1,表1),IHC的总感染效率为43.6%±13.5%,OHC的总感染效率为54.5%±为12.7%(与AAV2.7m8相比,IHC和OHC分别为p=0.003和0.03)。对于AAV8-GFP(n=4,图1,表1),IHC的总感染效率为86.0±5.34%,OHC的51.7%±5.95%(与AAV2.7m8相比,IHC和OHC分别为p=0.84和0.003)。
Anc80L65是据报道能感染IHCs和OHCs二者的合成AAV。当使用后管法将Anc80L65-GFP注射到新生小鼠内耳(n=7,图1,表1)时,IHC的总感染效率为94.0±3.20%,OHC的总感染效率为67.0±3.81%.尽管AAV2.7m8和Anc80L65之间的IHC感染效率相当(p=0.16),但这些数据显示,与Anc80L65相比,AAV2.7m8更能感染OHCs(p=0.04)。
详细检查接受AAV2.7m8-GFP注射的小鼠(n=8),显示AAV2.7m8能够感染整个耳蜗的毛细胞(图2a-2b)。耳蜗顶部的IHC感染效率为90.3±8.98%,中部转弯处的IHC感染效率为84.6±10.4%,耳蜗底部的IHC感染效率为77.5±10.8%。耳蜗顶部的OHC感染效率为89.0±9.53%,中部转弯处的OHC感染效率为85.2±10.9%,耳蜗底部的的OHC感染效率为74.9±12.2%。在注射AAV2.7m8的8只小鼠中,有4只在整个耳蜗中IHC和OHC感染效率超过90%(图2)。在注射AAV2.7m8的8只小鼠中,有1只的IHC和OHC感染效率低于30%。这可能反映了AAV2.7m8-GFP无意中进入外淋巴而不是内淋巴。总之,这些结果表明AAV2.7m8是能够高效地感染耳蜗IHCs和OHCs的强力病毒载体。
除了评估合成AAVs在耳蜗中的毛细胞感染效率外,还研究了前庭器官中的毛细胞感染效率。当将AAV2.7m8-GFP、AAV8BP2-GFP和AAV-DJ-GFP递送到新生小鼠内耳后,在前庭器官中表达了GFP。在胞囊中进行前庭毛细胞感染效率的定量(图3a-3g,表1)。AAV2.7m8-GFP的胞囊毛细胞感染效率为27.5±7.08%(n=8),AAV8BP2-GFP的胞囊毛细胞感染效率为34.2±6.77%(n=9,与AAV2.7m8细胞p=0.63),AAV-DJ-GFP的胞囊毛细胞感染效率为2.56±1.39%(n=5,与AAV2.7m8相比p=0.07)。还在新生小鼠胞囊中体内检验了AAV2-GFP、AAV8-GFP和Anc80L65-GFP的胞囊毛细胞感染效率(图3,表1)。AAV2的胞囊毛细胞感染效率为32.4±6.52%(n=3,与AAV2.7m8相比p=0.77),AAV8的胞囊毛细胞感染效率为93.3±1.77%(n=4,与AAV2.7m8相比p<.001),Anc80L65的胞囊毛细胞感染效率为67.7±2.68%(n=7,与AAV2.7m8相比p=0.002)。这些结果表明,AAV2.7m8以远高于前庭毛细胞的效率优先感染耳蜗毛细胞。
在耳蜗毛细胞作为内耳基因疗法研究中的靶细胞类型赢得最多关注的同时,围绕毛细胞的胶质样支持细胞也是基因疗法的重要靶点。支持细胞的一个特定亚群,即内柱细胞、内指细胞和第三排Deiters细胞,表达了富含亮氨酸的含重复序列G蛋白偶联受体5(LGR5),并表现出促进毛细胞再生的祖细胞样特性。当将AAV2.7m8-GFP递送到新生小鼠内耳时,在两种LGR5+支持细胞类型中看到了GFP表达-内柱细胞和内指细胞(图4,图5a-5g,表1)。总内柱细胞感染效率为86.1±4.87%(顶部为94.7±3.11%,中间转弯处为91.3±3.80%,底部为72.4±7.93%,n=8)。总内指细胞感染效率为61.4±9.30%(顶部为72.0±12.5%,中间转弯处为60.0±11.1%,底部为52.3±12.9%,n=4)。相比之下,注射AAV8BP2的小鼠在内柱细胞和内指细胞无GFP表达(图4a-4g)。在注射以下物质的小鼠中也看到内柱细胞感染:AAV-DJ-GFP(10.9±3.67%,n=5,与AAV2.7m8相比p<.001),AAV2-GFP(60.3±7.96%,n=3,与AAV2.7m8相比p=0.007),AAV8-GFP(50.4±8.64%,n=4,与AAV2.7m8相比p<.001),和Anc80L65-GFP(75.3±4.94%,n=7,p=0.11)。然而,这些AAVs均不感染内指细胞。这些结果表明,AAV2.7m8能够感染被认为能高效促进毛细胞再生的支持细胞亚群(内柱细胞和内指细胞)。
为使内耳基因疗法成为治疗听力损失和前庭功能障碍的可行方法,所使用的病毒载体应该对正常的听觉和前庭功能产生最小的影响。为评估合成AAVs的内耳递送是否对听力有任何影响,我们测量了听性脑干反应(ABRs)(图6a-6b)。与未经历内耳操作的对照小鼠相比,经历AAV2.7m8-GFP(n=8),AAV-DJ-GFP(n=5),AAV2-GFP(n=3),AAV8-GFP(n=4),和Anc80L65-GFP(n=7)注射的小鼠显示ABR阈值无显著变化(分别为p=0.09,0.11,0.25,0.43,和0.25,ANOVA)。相比之下,与对照小鼠相比,经历AAV8BP2-GFP(n=13)注射的小鼠显示10-25dB的ABR阈值升高(p<.001,ANOVA)。使用Scheffe方法进行的事后比较显示,在4kHz、8kHz、16kHz和32kHz时,ABR阈值差异具有统计学显著性(分别为p=0.004、<0.001、<0.001和0.034)。有可能是AAV8BP2对小鼠内耳的免疫原性更强,这导致耳蜗毛细胞丢失(图1)以及ABR阈值升高。AAV8BP2注射后的耳蜗检查显示了炎症细胞浸润(图7a-7b)。当以初始浓度的一半(0.5x1010 G.C.)注射AAV8BP2-GFP时,ABR阈值与对照小鼠相当(p=0.49,图8),但IHC和OHC感染效率也降低了(43.2%±8.36%和23.3%±5.41%,n=5),尽管该变化在统计学上不是显著的(IHC和OHC分别为p=0.38和0.08)。
患有前庭功能障碍的小鼠经常会表现出转圈行为。为评估合成AAV的内耳递送是否对前庭***有任何影响,检查了注射小鼠的转圈行为(图6)。未经历内耳基因递送的对照小鼠每2分钟转圈5.11±0.78次(n=6)。用以下物质注射的小鼠的转圈行为类似于未注射对照小鼠(分别为p=0.92,0.05,0.31,0.28,和0.60,ANOVA):AAV2.7m8-GFP(5.04±0.54次/2分钟,n=8),AAV-DJ-GFP(6.20±0.36次/2分钟,n=5),AAV2-GFP(6.00±1.02次/2分钟,n=3),AAV8-GFP(4.58±0.28次/2分钟,n=4),和Anc80L65-GFP(5.52±0.65次/2分钟,n=7)。相比之下,经历AAV8BP2-GFP注射的小鼠的转圈稍有增加(6.87±0.38次/2分钟,p=0.009,n=13)。与对照动物相比,以初始剂量的一半(0.5x1010 G.C.)注射AAV8BP2-GFP不导致转圈行为增加(5.47±0.77次/2分钟,p=0.66,n=5,图8a-8c)。这些结果表明,AAV2.7m8内耳递送是安全的,对听觉和前庭功能的不良作用很小。
虽然有几项研究表明病毒内耳基因疗法可改善遗传性听力损失小鼠模型的听力功能,但听力恢复通常是不完全的(Emptoz,A.等人,Local gene therapy durablyrestores vestibular function in a mouse model of Usher syndrome type 1G.ProcNatl Acad Sci U S A 114,9695-9700(2017))。传统AAV的主要缺点之一是其感染OHCs的效率低。我们的结果表明,AAV2.7m8能够高效地感染耳蜗IHCs和OHCs。事实上,当通过后管方法递送时,AAV2.7m8感染OHCs的效率甚至比Anc80L65更高。发现与前庭毛细胞相比,AAV2.7m8更优先靶向耳蜗毛细胞。这与同样高效地感染前庭毛细胞的Anc80L65不同(Landegger,L.D.等人,A synthetic AAV vector enables safe and efficient genetransfer to the mammalian inner ear.Nat Biotechnol 35,280-284(2017))。AAV2.7m8靶向耳蜗毛细胞的偏好可用于仅需要在耳蜗中进行转基因表达的研究中,其可以潜在地最小化来自前庭***中不需要的转基因表达引起的前庭毒性。
大多数内耳基因疗法研究都集中在遗传性耳聋的动物模型上。然而,遗传性听力损失的患病率远低于其他类型的听力损失,如年龄相关的听力损失(老年性耳聋)和噪声诱导的听力损失。应用基因疗法治疗老年性耳聋和噪声诱导的听力损失的策略之一是诱导毛细胞再生。非哺乳动物(如鸟类和斑马鱼)的毛细胞在损伤后会再生,而哺乳动物的毛细胞则不会再生。支持细胞被认为是毛细胞再生的来源。在哺乳动物内耳中,支持细胞亚群LGR5+(内柱细胞、内指细胞和第三排Deiters细胞)具有促进毛细胞再生的祖细胞样特性。为利用基因疗法诱导毛细胞再生,一个关键的因素是要有能够有效靶向此支持细胞群的病毒载体。如本文所示,AAV2.7m8有效地感染耳蜗内毛细胞和外毛细胞二者。此外,它还以非常高的效率感染已被其他人证明为LGR5+(内柱细胞和内指细胞)的支持细胞类型。综上所述,AAV2.7m8是一种强效的病毒载体,可以极大地拓展内耳基因疗法的应用领域。
序列表
<110> Chien, Wade
Bennett, Jean
<120> 腺相关病毒及其用于内耳治疗的用途
<130> 077867-1171567
<150> US 62/784,306
<151> 2018-12-21
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 合成构建物
<400> 1
Leu Gly Glu Thr Thr Arg Pro
1 5
<210> 2
<211> 735
<212> PRT
<213> 人工序列
<220>
<223> 合成构建物
<400> 2
Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser
1 5 10 15
Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro Gly Pro Pro Pro Pro
20 25 30
Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro
35 40 45
Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro
50 55 60
Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp
65 70 75 80
Arg Gln Leu Asp Ser Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala
85 90 95
Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly
100 105 110
Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro
115 120 125
Leu Gly Leu Val Glu Glu Pro Val Lys Thr Ala Pro Gly Lys Lys Arg
130 135 140
Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr Gly
145 150 155 160
Lys Ala Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr
165 170 175
Gly Asp Ala Asp Ser Val Pro Asp Pro Gln Pro Leu Gly Gln Pro Pro
180 185 190
Ala Ala Pro Ser Gly Leu Gly Thr Asn Thr Met Ala Thr Gly Ser Gly
195 200 205
Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser
210 215 220
Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly Asp Arg Val Ile
225 230 235 240
Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu
245 250 255
Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr
260 265 270
Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His
275 280 285
Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp
290 295 300
Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln Val
305 310 315 320
Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu
325 330 335
Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr
340 345 350
Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp
355 360 365
Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser
370 375 380
Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser
385 390 395 400
Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe Glu
405 410 415
Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg
420 425 430
Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr
435 440 445
Asn Thr Pro Ser Gly Thr Thr Thr Gln Ser Arg Leu Gln Phe Ser Gln
450 455 460
Ala Gly Ala Ser Asp Ile Arg Asp Gln Ser Arg Asn Trp Leu Pro Gly
465 470 475 480
Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser Ala Asp Asn Asn
485 490 495
Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly
500 505 510
Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys Asp
515 520 525
Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe Gly Lys
530 535 540
Gln Gly Ser Glu Lys Thr Asn Val Asp Ile Glu Lys Val Met Ile Thr
545 550 555 560
Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr
565 570 575
Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Arg Gln Ala Ala Thr
580 585 590
Ala Asp Val Asn Thr Gln Gly Val Leu Pro Gly Met Val Trp Gln Asp
595 600 605
Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr
610 615 620
Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys
625 630 635 640
His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asn
645 650 655
Pro Ser Thr Thr Phe Ser Ala Ala Lys Phe Ala Ser Phe Ile Thr Gln
660 665 670
Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys
675 680 685
Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr
690 695 700
Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr
705 710 715 720
Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu
725 730 735
Claims (25)
1.重组腺相关病毒(AAV)病毒粒子,包含:
a)修饰的AAV衣壳蛋白,其中所述修饰的AAV衣壳蛋白相对于相应的亲本AAV衣壳蛋白包含肽***,其中所述肽***包含氨基酸序列LGETTRP(SEQ ID NO:1),其中所述修饰的AAV衣壳蛋白中的***在对应于AAV2-VP1的氨基酸587和588的氨基酸之间;以及
b)产生表达产物的异源核酸,其中所述表达产物减少听力损失或眩晕。
2.权利要求1的重组AAV病毒粒子,其中所述表达产物是降低与听力损失相关基因的表达的核酸,其中所述基因选自下组:DIAPH1、KCNQ4、GJB3、IFNLR1、GJB2、GJB6、MYH1、CEACAM16、GSDME/DFNA5、WFS1、LMX1A、TECTA、COCH、EYA4、MYO7A、COL11A2、POU4F3、MYH9、ACTG1、MYO6、SIX1、SLC17A8、REST、GRHL2、NLRP3、TMC1、COL11A1、CRYM、P2RX2、CCDC50、MIRN96、TJP2、TNC、SMAC/DIABLO、TBC1D24、CD164、OSBPL2、HOMER2、KITLG、MCM2、PTPRQ、DMXL2、MYO3A和PDE1C。
3.权利要求1的重组AAV病毒粒子,其中所述表达产物是减少听力损失的多肽,其中所述多肽选自下组:GJB2、GJB6、MYO7A、MYO15A、SLC26A4、TMIE、TMC1、TMPRSS3、OTOF、CDH23、GIPC3、STRC、USH1C、OTOG、TECTA、OTOA、PCDH15、RDX、GRXCR1、TRIOBP、CLDN14、MYO3A、WHRN、CDC14A、ESRRB、ESPN、MYO6、HGF、ILDR1、ADCY1、CIB2、MARVELD2、BDP1、COL11A2、PDZD7、PJVK、SLC22A4、SLC26A5、LRTOMT/COMT2、DCDC2、LHFPL5、S1PR2、PNPT1、BSND、MSRB3、SYNE4、LOXHD1、TPRN、GPSM2、PTPRQ、OTOGL、TBC1D24、ELMOD3、KARS、SERPINB6、CABP2、NARS2、MET、TSPEAR、TMEM132E、PPIP5K2、GRXCR2、EPS8、CLIC5、FAM65B、DFNB32、EPS8L2、ROR1、WBP2、ESRP1、MPZL2、PRPS1、POU3F4、SMPX、AIFM1和COL4A。
4.权利要求1-3中任一项的重组AAV病毒粒子,其中所述AAV病毒粒子是AAV2病毒粒子、AAV5病毒粒子、AAV8病毒粒子或AAV9病毒粒子。
5.权利要求1-4中任一项的重组AAV病毒粒子,其中所述AAV病毒粒子是AAV2.7m8病毒粒子。
6.权利要求2-5中任一项的重组AAV病毒粒子,其中减少与听力损失相关的基因表达的所述核酸是干扰RNA。
7.权利要求6的重组AAV病毒粒子,其中所述干扰RNA为反义分子、短干扰RNA或miRNA。
8.权利要求中1-7中任一项的重组AAV病毒粒子,其中所述听力损失选自下组:年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失和外伤所致的听力损失。
9.用于治疗或预防受试者的内耳毛细胞损伤的方法,包括向具有内耳毛细胞损伤或有风险发生内耳毛细胞损伤的受试者施用有效量的权利要求1-8中任一项的重组AAV病毒粒子。
10.权利要求9的方法,其中所述受试者具有或有风险发生年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失和外伤所致的听力损失。
11.权利要求9或10的方法,其中所述重组AAV病毒粒子感染耳蜗的内毛细胞和外毛细胞。
12.权利要求9-11中任一项的方法,其中所述重组AAV病毒粒子感染耳蜗中的胶质样支持细胞。
13.权利要求12的方法,其中所述支持细胞为内柱细胞或内指细胞。
14.权利要求9-13中任一项的方法,其中所述重组AAV病毒粒子增加内耳毛细胞再生。
15.权利要求14的方法,其中所述重组AAV病毒粒子增加耳蜗毛细胞再生。
16.用于治疗或预防受试者的听力损失或眩晕的方法,包括向具有听力损失或眩晕或者有风险发生听力损失或眩晕的受试者施用有效量的权利要求1-8中任一项的重组AAV病毒粒子。
17.权利要求16的方法,其中所述受试者具有或有风险发生年龄相关的听力损失、遗传性听力损失、噪声诱导的听力损失、疾病相关性听力损失和外伤所致的听力损失。
18.权利要求16或17的方法,其中所述重组AAV病毒粒子感染所述受试者的内耳毛细胞。
19.权利要求18的方法,其中所述内耳毛细胞是耳蜗的内毛细胞和外毛细胞。
20.权利要求19中任一项的方法,其中所述重组AAV病毒粒子感染耳蜗中的胶质样支持细胞。
21.权利要求20的方法,其中所述支持细胞为内柱细胞或内指细胞。
22.权利要求16-21中任一项的方法,其中所述重组AAV病毒粒子增加内耳毛细胞再生。
23.权利要求22的方法,其中所述重组AAV病毒粒子增加耳蜗毛细胞再生。
24.权利要求9-22中任一项的方法,其中所述重组AAV病毒粒子经静脉内施用,经鞘内施用,经鼓膜内(intratypmanically)施用,经圆窗施用,经半规管递送施用,或经镫骨足板造孔术施用。
25.权利要求24的方法,其中所述重组AAV病毒粒子经由经管造口术施用到所述受试者的后半规管中。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862784306P | 2018-12-21 | 2018-12-21 | |
US62/784,306 | 2018-12-21 | ||
PCT/US2019/068070 WO2020132578A1 (en) | 2018-12-21 | 2019-12-20 | Adeno-associated viruses and their uses for inner ear therapy |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113825521A true CN113825521A (zh) | 2021-12-21 |
Family
ID=69185733
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201980085229.0A Pending CN113825521A (zh) | 2018-12-21 | 2019-12-20 | 腺相关病毒及其用于内耳疗法的用途 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20220096658A1 (zh) |
EP (1) | EP3897679A1 (zh) |
CN (1) | CN113825521A (zh) |
IL (1) | IL284099A (zh) |
WO (1) | WO2020132578A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2021150850A1 (en) * | 2020-01-24 | 2021-07-29 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Svcs. | Adeno-associated viruses and their uses for inner ear therapy |
US20230181763A1 (en) * | 2020-04-30 | 2023-06-15 | The United States Of America,As Represented By The Secretary,Department Of Health And Human Services | Application of aav44.9 vector in gene therapy for the inner ear |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180066022A1 (en) * | 2015-03-02 | 2018-03-08 | Adverum Biotechnologies, Inc. | Compositions and methods for intravitreal delivery of polynucleotides to retinal cones |
WO2018145111A1 (en) * | 2017-02-06 | 2018-08-09 | Children's Medical Center Corporation | Materials and methods for delivering nucleic acids to cochlear and vestibular cells |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
RS60207B1 (sr) | 2011-04-22 | 2020-06-30 | Univ California | Adeno-povezani virioni virusa sa varijantama kapsida i postupci za njihovu primenu |
-
2019
- 2019-12-20 CN CN201980085229.0A patent/CN113825521A/zh active Pending
- 2019-12-20 US US17/416,311 patent/US20220096658A1/en active Pending
- 2019-12-20 EP EP19839761.4A patent/EP3897679A1/en active Pending
- 2019-12-20 WO PCT/US2019/068070 patent/WO2020132578A1/en unknown
-
2021
- 2021-06-16 IL IL284099A patent/IL284099A/en unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180066022A1 (en) * | 2015-03-02 | 2018-03-08 | Adverum Biotechnologies, Inc. | Compositions and methods for intravitreal delivery of polynucleotides to retinal cones |
WO2018145111A1 (en) * | 2017-02-06 | 2018-08-09 | Children's Medical Center Corporation | Materials and methods for delivering nucleic acids to cochlear and vestibular cells |
Non-Patent Citations (1)
Title |
---|
HANEN KHABOU ET AL.: "Insight into the mechanisms of enhanced retinal transduction by the engineered AAV2 capsid variant-7m8", BIOTECHNOLOGY AND BIOENGINEERING, vol. 113, no. 12, 31 December 2016 (2016-12-31), pages 2 - 7, XP055832410, DOI: 10.1002/bit.26031 * |
Also Published As
Publication number | Publication date |
---|---|
US20220096658A1 (en) | 2022-03-31 |
IL284099A (en) | 2021-08-31 |
EP3897679A1 (en) | 2021-10-27 |
WO2020132578A1 (en) | 2020-06-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109310745B (zh) | 用于将核酸递送至耳蜗和前庭细胞的材料和方法 | |
Luoni et al. | Whole brain delivery of an instability-prone Mecp2 transgene improves behavioral and molecular pathological defects in mouse models of Rett syndrome | |
JP5766389B2 (ja) | 脊髄疾患の遺伝子療法 | |
AU2018261003A2 (en) | Compositions and methods of treating Huntington's Disease | |
TW201905200A (zh) | 調節多核苷酸 | |
JP7430652B2 (ja) | 蝸牛および前庭細胞に核酸を送達するための組成物および方法 | |
TW201928060A (zh) | 肌萎縮性脊髓側索硬化症(als)之治療 | |
US20220125875A1 (en) | Aav-mediated gene therapy restoring the otoferlin gene | |
JP2019533428A (ja) | 標的遺伝子導入のための方法および組成物 | |
JP2020513831A (ja) | MeCP2発現カセット | |
US20220347321A1 (en) | Expression of neuropeptides | |
CN113825521A (zh) | 腺相关病毒及其用于内耳疗法的用途 | |
EP3634986A1 (en) | Gene therapy for ocular disorders | |
US20220348957A1 (en) | Adeno-associated viruses and their uses for inner ear therapy | |
WO2010037143A1 (en) | Vectors and methods of treating brain seizures | |
JP2023529503A (ja) | 網膜症のためのaav媒介性遺伝子導入 | |
WO2023143366A1 (zh) | 变异型腺相关病毒及其在疾病治疗中的应用 | |
JP7504967B2 (ja) | MeCP2発現カセット | |
US20220119475A1 (en) | Recombinant adeno associated virus encoding clarin-1 and uses thereof | |
KR20240053630A (ko) | 오토페를린의 이소형 5를 인코딩하는 이중 재조합 aav8 벡터 시스템 및 이의 용도 | |
Cao | Application of Gene Editing to Promote Axon Regeneration in Retinal Ganglion Cells after Optic Nerve Injury | |
EP3934699A1 (en) | Neuroprotective gene therapy targeting the akt pathway | |
CN117866970A (zh) | 用于无脉络膜症的基因工程构建体 | |
TW202413649A (zh) | 肌萎縮性脊髓側索硬化症(als)之治療 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |