HRP20191233T1 - Antitijela lakog lanca modificirana histidinom i genetski modificirane nehumane životinje za dobivanje istih - Google Patents
Antitijela lakog lanca modificirana histidinom i genetski modificirane nehumane životinje za dobivanje istih Download PDFInfo
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- HRP20191233T1 HRP20191233T1 HRP20191233TT HRP20191233T HRP20191233T1 HR P20191233 T1 HRP20191233 T1 HR P20191233T1 HR P20191233T T HRP20191233T T HR P20191233TT HR P20191233 T HRP20191233 T HR P20191233T HR P20191233 T1 HRP20191233 T1 HR P20191233T1
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- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 title claims 7
- 108090000623 proteins and genes Proteins 0.000 claims 37
- 235000014304 histidine Nutrition 0.000 claims 21
- 241000283984 Rodentia Species 0.000 claims 20
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 claims 17
- 238000006467 substitution reaction Methods 0.000 claims 17
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 claims 16
- 150000002411 histidines Chemical class 0.000 claims 15
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 claims 11
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 claims 11
- 239000000427 antigen Substances 0.000 claims 7
- 108091007433 antigens Proteins 0.000 claims 7
- 102000036639 antigens Human genes 0.000 claims 7
- 108020004705 Codon Proteins 0.000 claims 6
- 210000004602 germ cell Anatomy 0.000 claims 6
- 238000000034 method Methods 0.000 claims 4
- 230000008707 rearrangement Effects 0.000 claims 4
- 101001008255 Homo sapiens Immunoglobulin kappa variable 1D-8 Proteins 0.000 claims 2
- 101001047628 Homo sapiens Immunoglobulin kappa variable 2-29 Proteins 0.000 claims 2
- 101001008321 Homo sapiens Immunoglobulin kappa variable 2D-26 Proteins 0.000 claims 2
- 101001047619 Homo sapiens Immunoglobulin kappa variable 3-20 Proteins 0.000 claims 2
- 101001008263 Homo sapiens Immunoglobulin kappa variable 3D-15 Proteins 0.000 claims 2
- 102100022949 Immunoglobulin kappa variable 2-29 Human genes 0.000 claims 2
- 230000002378 acidificating effect Effects 0.000 claims 2
- 230000007935 neutral effect Effects 0.000 claims 2
- 108060003951 Immunoglobulin Proteins 0.000 claims 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 claims 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 claims 1
- 210000003719 b-lymphocyte Anatomy 0.000 claims 1
- 230000001747 exhibiting effect Effects 0.000 claims 1
- 230000003053 immunization Effects 0.000 claims 1
- 102000018358 immunoglobulin Human genes 0.000 claims 1
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- A—HUMAN NECESSITIES
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- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0278—Knock-in vertebrates, e.g. humanised vertebrates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
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- 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
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/15—Humanized animals
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/072—Animals genetically altered by homologous recombination maintaining or altering function, i.e. knock in
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/15—Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
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- C07K2317/565—Complementarity determining region [CDR]
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- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
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Claims (12)
1. Genetski modificirani glodavac koji u svojoj zametnoj liniji sadrži lokus imunoglobulinskog lakog lanca koji sadrži dva nepreuređena humana segmenta gena Vκ i jedan ili više nepreuređenih humanih segmenata gena Jκ operativno povezanih sa sekvencom konstantne regije imunoglobulinskog lakog lanca,
naznačeno time, što su nepreuređeni humani segmenti gena Vκ humani segmenti gena Vκ1-39 i Vκ3-20, od kojih svaki sadrži jednu ili više supstitucija nehistidinskog CDR3 kodona histidinskim kodonom,
naznačeno time, što su humani segmenti gena Vκ i Jκ sposobni za preuređenje, a humani segmenti gena Vκ i Jκ šifriraju varijabilnu domenu humanog lakog lanca koja sadrži jedan ili više histidina, pri čemu su jedan ili više histidina dobiveni iz jedne ili više supstitucija,
naznačeno time, što je (i) supstitucija jednog ili više histidna humanog segmenta gena Vκ1-39 te je supstitucija dizajnirana za izražavanje jednog ili više histidina na poziciji odabranoj iz skupine koja se sastoji od 105, 106, 108 i 111 humanog segmenta gena Vκ1-39 (prema numeriranju IMGT) i njihove kombinacije; i/ili (ii) supstitucija jednog ili više histidina humanog segmenta gena Vκ3-20 te je supstitucija dizajnirana za izražavanje jednog ili više histidina na poziciji odabranoj iz skupine koja se sastoji od 105, 106, 107 i 109 humanog segmenta gena Vκ3-20 (prema numeriranju IMGT) i njihove kombinacije, i
naznačeno time, što glodavac dodatno u svojoj zametnoj liniji sadrži lokus imunoglobulinskog teškog lanca koji sadrži nepreuređenu sekvencu varijabilne regije imunoglobulinskog teškog lanca koja sadrži humane segmente gena VH, DH i JH operativno povezane sa sekvencom konstantne regije imunoglobulinskog teškog lanca.
2. Glodavac prema patentnom zahtjevu 1, naznačen time, što:
(i) glodavac ne sadrži endogeni segment gena varijabilne regije lakog lanca κ koji je sposoban za preuređenje kako bi formirao imunoglobulinski laki lanac;
(ii) sekvenca konstantne regije imunoglobulinskog lakog lanca je nehumana sekvenca konstantne regije imunoglobulinskog lakog lanca, opcionalno naznačena time, što je nehumana sekvenca konstantne regije imunoglobulinskog lakog lanca mišja ili štakorska sekvenca ili što je nehumana sekvenca konstantne regije imunoglobulinskog lakog lanca endogena sekvenca konstantne regije imunoglobulinskog lakog lanca;
(iii) sekvenca konstantne regije imunoglobulinskog teškog lanca je nehumana sekvenca konstantne regije imunoglobulinskog teškog lanca, kao što je mišja ili štakorska sekvenca;
(iii) dva nepreuređena humana segmenta gena Vκ i jedan ili više nepreuređenih humanih segmenata gena Jκ su prisutni na endogenom lokusu imunoglobulinskog lakog lanca; ili
(iv) konstantna regija imunoglobulinskog lakog lanca je Cκ regija.
3. Glodavac prema patentnom zahtjevu 1, naznačen time, što je nehumana sekvenca konstantne regije imunoglobulinskog teškog lanca endogena nehumana sekvenca konstantne regije imunoglobulinskog teškog lanca.
4. Glodavac prema patentnom zahtjevu 1, naznačen time, što je glodavac štakor ili miš.
5. Glodavac prema patentnom zahtjevu 1, naznačen time, što glodavac sadrži populaciju B stanica kao odgovor na interesni antigen koja je obogaćen za antitijela koja izražavaju smanjenje disocijativnog poluživota (t1/2) pri kiseloj pH vrijednosti u usporedbi s neutralnom pH vrijednošću od najmanje 2 puta, najmanje 3 puta, najmanje 4 puta, najmanje 5 puta, najmanje 10 puta, najmanje 15 puta, najmanje 20 puta, najmanje 25 puta ili najmanje 30 puta.
6. Glodavac prema patentnom zahtjevu 1, naznačen time, što glodavac sadrži pet humanih segmenata Jκ, a tih pet humanih segmenata Jκ su segmenti Jκ1, Jκ2, Jκ3, Jκ4 i Jκ5.
7. Glodavac prema patentnom zahtjevu 1, naznačen time, što lokus imunoglobulinskog lakog lanca ne sadrži više od humanih segmenata gena Vκ1-39 i Vκ3-20 i jedan ili više nepreuređenih humanih segmenata gena Jκ koji su operativno povezani sa sekvencom konstantne regije imunoglobulinskog lakog lanca.
8. Glodavac prema patentnom zahtjevu 7, naznačen time, što svaki nepreuređeni humani segment gena Vκ1-39 i Vκ3-20 sadrži supstituciju tri ili četiri nehistidinska CDR3 kodona histidinskim kodonima, opcionalno naznačen time, što:
(i) supstitucija humanog gena Vk1-39 je dizajnirana za izražavanje histidina na pozicijama 106, 108 i 111 humanog segmenta gena Vκ1-39;
(ii) supstitucija humanog gena Vκ1-39 je dizajnirana za izražavanje histidina na položajima 105, 106, 108 i 111 humanog segmenta gena Vκ1-39;
(iii) supstitucija humanog segmenta gena Vκ3-20 je dizajnirana za izražavanje histidina na položajima 105, 106, i 109 humanog segmenta gena Vκ3-20;
(iv) supstitucija humanog segmenta gena Vκ3-20 je dizajnirana za izražavanje histidina na položajima 105, 106, 107 i 109 humanog segmenta gena Vκ3-20.
9. Glodavac prema patentnom zahtjevu 1, naznačen time, što glodavac izražava populaciju antitijela specifičnih za antigen kao odgovor na antigen, naznačen time, što sva antitijela u populaciji sadrže:
varijabilne domene imunoglobulinskog lakog lanca dobivene iz preuređivanja nepreuređenih humanih segmenata gena Vκ1-39 i Vκ3-20 i jednog ili više nepreuređenih humanih segmenata Jκ, i
imunoglobulinske teške lance koji sadrže humane varijabilne domene teškog lanca dobivene iz niza humanih teških segmenata V, D i J.
10. Metoda za dobivanje glodavca koji sadrži genetski modificirani lokus imunoglobulinskog lakog lanca u svojoj zametnoj liniji, pri čemu se metoda sastoji od:
modificiranja genoma zametne linije glodavca radi brisanja ili postizanja nefunkcionalnosti endogenih segmenata gena Vκ i Jκ imunoglobulinskog lakog lanca u lokusu imunoglobulinskog lakog lanca, i
umetanja u genom zametne linije nehumane životinje varijabilne regije imunoglobulinskog lakog lanca koja sadrži dva nepreuređena humana segmenta gena V Vκ i najmanje jedan nepreuređeni humani segment gena Jκ, tako da je sekvenca varijabilne regije imunoglobulinskog lakog lanca operativno povezana sa sekvencom konstantne regije imunoglobulina,
naznačeno time, što su dva nepreuređena humana segmenta gena Vκ humani segmenti gena Vκ1-39 i Vκ3-20, od kojih svaki sadrži jednu ili više supstitucija nehistidinskog CDR3 kodona histidinskim kodonom,
naznačeno time, što su nepreuređeni humani segmenti gena Vκ i Jκ sposobni za sposobni za preuređenje, a nepreuređeni humani segmenti gena Vκ i Jκ šifriraju varijabilnu domenu humanog lakog lanca koja sadrži jedan ili više histidina koji su dobiveni iz jedne ili više supstitucija,
naznačeno time, što je (i) supstitucija jednog ili više histidna humanog segmenta gena Vκ1-39 te je supstitucija dizajnirana za izražavanje jednog ili više histidina na poziciji odabranoj iz skupine koja se sastoji od 105, 106, 108 i 111 humanog segmenta gena Vκ1-39 (prema numeriranju IMGT) i njihove kombinacije; i/ili (ii) supstitucija jednog ili više histidina humanog segmenta gena Vκ3-20 te je supstitucija dizajnirana za izražavanje jednog ili više histidina na poziciji odabranoj iz skupine koja se sastoji od 105, 106, 107 i 109 humanog segmenta gena Vκ3-20 (prema numeriranju IMGT) i njhove kombinacije, i
naznačeno time, što glodavac dodatno u svojoj zametnoj liniji sadrži lokus imunoglobulinskog teškog lanca koji sadrži nepreuređenu sekvencu varijabilne regije imunoglobulinskog teškog lanca koja sadrži humane segmente gena VH, DH i JH operativno povezane sa sekvencom konstantne regije imunoglobulinskog teškog lanca.
11. Metoda prema patentnom zahtjevu 10, naznačena time, što:
(i) varijabilna regija imunoglobulinskog lakog lanca se nalazi na endogenom nehumanom lokusu imunoglobulinskog lakog lanca; ili
(ii) glodavac je miš ili štakor.
12. Metoda za dobivanje antitijela koje izražava vezivanje za interesni antigen ovisno o pH vrijednosti koja se sastoji od:
imunizacije glodavca prema patentnom zahtjevu 1 interesnim antigenom te odabira antitijela koje se vezuje za interesni antigen sa željenim afinitetom pri neutralnoj pH vrijednosti uz istovremeno ispoljavanje smanjenog vezivanja za interesni antigen pri kiseloj pH vrijednosti.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US14/030,424 US20140013456A1 (en) | 2012-03-16 | 2013-09-18 | Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same |
| PCT/US2014/056285 WO2015042250A1 (en) | 2013-09-18 | 2014-09-18 | Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same |
| EP14781761.3A EP3046412B1 (en) | 2013-09-18 | 2014-09-18 | Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same |
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| HRP20191233T1 true HRP20191233T1 (hr) | 2019-10-18 |
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| HRP20191233TT HRP20191233T1 (hr) | 2013-09-18 | 2019-07-10 | Antitijela lakog lanca modificirana histidinom i genetski modificirane nehumane životinje za dobivanje istih |
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| JP (3) | JP6621750B2 (hr) |
| KR (2) | KR102150414B1 (hr) |
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| BR (1) | BR112016005912A2 (hr) |
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| DK (1) | DK3046412T3 (hr) |
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| KR102147548B1 (ko) | 2011-02-25 | 2020-08-24 | 추가이 세이야쿠 가부시키가이샤 | ＦｃγＲＩＩｂ 특이적 Ｆｃ 항체 |
| WO2013047748A1 (ja) | 2011-09-30 | 2013-04-04 | 中外製薬株式会社 | 複数の生理活性を有する抗原の消失を促進する抗原結合分子 |
| WO2013081143A1 (ja) | 2011-11-30 | 2013-06-06 | 中外製薬株式会社 | 免疫複合体を形成する細胞内への運搬体（キャリア）を含む医薬 |
| TW202237660A (zh) | 2012-08-24 | 2022-10-01 | 日商中外製藥股份有限公司 | FcγRIIb特異性Fc區域變異體 |
| CN105102618B (zh) | 2012-12-27 | 2018-04-17 | 中外制药株式会社 | 异源二聚化多肽 |
| CN113621057A (zh) | 2013-04-02 | 2021-11-09 | 中外制药株式会社 | Fc区变体 |
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