RU2224764C2 - Модифицированный полипептид с увеличенным временем полужизни, его получение и использование - Google Patents
Модифицированный полипептид с увеличенным временем полужизни, его получение и использование Download PDFInfo
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- 229920001184 polypeptide Polymers 0.000 title claims abstract 53
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract 53
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract 53
- 102000005962 receptors Human genes 0.000 claims abstract 15
- 108020003175 receptors Proteins 0.000 claims abstract 15
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 8
- 238000000034 method Methods 0.000 claims abstract 6
- 102100022339 Integrin alpha-L Human genes 0.000 claims abstract 4
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 claims abstract 4
- 210000003734 kidney Anatomy 0.000 claims abstract 3
- 230000001404 mediated effect Effects 0.000 claims abstract 3
- 241000124008 Mammalia Species 0.000 claims abstract 2
- 238000001727 in vivo Methods 0.000 claims 6
- 239000001963 growth medium Substances 0.000 claims 3
- 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 claims 2
- 108020004414 DNA Proteins 0.000 claims 2
- 102000053602 DNA Human genes 0.000 claims 2
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 claims 2
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 239000005557 antagonist Substances 0.000 claims 1
- 230000004071 biological effect Effects 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 238000002703 mutagenesis Methods 0.000 claims 1
- 231100000350 mutagenesis Toxicity 0.000 claims 1
- 108020004707 nucleic acids Proteins 0.000 claims 1
- 102000039446 nucleic acids Human genes 0.000 claims 1
- 150000007523 nucleic acids Chemical class 0.000 claims 1
- 238000010276 construction Methods 0.000 abstract 1
- 230000006735 deficit Effects 0.000 abstract 1
- 230000000694 effects Effects 0.000 abstract 1
- 230000007721 medicinal effect Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
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Abstract
Изобретение относится к биотехнологии, может быть использовано в медицинской практике и касается полипептида, который выводится через почки и не содержит в своей исходной форме Fc-области IgG. Исходный полипептид модифицируют так, чтобы включить спасательный рецепторсвязывающий эпитоп Fc-области IgG и посредством этого увеличить циркуляторное время полужизни. Модифицированный полипептид содержит аминокислотную последовательность Ig-домена или Ig-подобного, который не является СН2-доменом, и аминокислотную последовательность спасательного рецепторсвязывающего эпитопа Fc-области IgG, расположенную в пределах Ig-домена или Ig-подобного домена. Аминокислотная последовательность модифицированного полипептида, по меньшей мере, на 70% идентична таковой исходного полипептида. Раскрыт способ получения указанного полипептида с использованием ДНК (в конструкции вектора), кодирующей указанный полипептид. Полученный полипептид может использоваться для лечения опосредованного LFA-1 нарушения у млекопитающих, в том числе человека. Изобретение позволяет увеличить время полужизни полипептида, за счет чего повышается эффективность лечения. 8 с. и 17 з.п. ф-лы, 4 ил., 3 табл.
Description
Текст описания в факсимильном виде (см. графическую часть)н
Claims (25)
1. Модифицированный полипептид, обладающий увеличенной продолжительностью времени полужизни in vivo и характеризующийся тем, что он получен на основе представляющего интерес полипептида, который не содержит Fc-область IgG и выводится через почки, причем аминокислотная последовательность представляющего интерес полипептида модифицирована таким образом, что она содержит а) аминокислотную последовательность Ig-домена или Ig-подобного домена, который не является СН2-доменом, и б) аминокислотную последовательность спасательного рецепторсвязывающего эпитопа Fc-области IgG, которая расположена в пределах Ig-домена или Ig-подобного домена, где аминокислотная последовательность модифицированного полипептида, по меньшей мере, на 70% идентична аминокислотной последовательности представляющего интерес полипептида и его биологическая активность сохраняется.
2. Модифицированный полипептид по п.1, отличающийся тем, что Ig-домен или Ig-подобный домен содержит домен CH1.
3. Модифицированный полипептид по п.1, отличающийся тем, что спасательный рецепторсвязывающий эпитоп происходит из одной или двух петель Fc-области и расположен в Ig-домене или Ig-подобном домене.
4. Модифицированный полипептид по п.1, отличающийся тем, что спасательный рецепторсвязывающий эпитоп происходит из СН2-домена Fc-области, модифицированный полипептид дополнительно включает CH1-, СН3- или VH-область и указанный эпитоп расположен в CH1-, СН3- или Vн-области или в нескольких таких областях Ig-домена или Ig-подобного домена.
5. Модифицированный полипептид по п.1, отличающийся тем, что спасательный рецепторсвязывающий эпитоп происходит из СН2-домена Fc-области и расположен в Ig-домене или Ig-подобном домене.
6. Модифицированный полипептид по п.1, отличающийся тем, что представляющий интерес полипептид является Fab, (Fab’)2, диантителом, Fv-фрагментом, одноцепочечным Fv-фрагментом или рецептором.
7. Модифицированный полипептид по п.1, отличающийся тем, что представляющий интерес полипептид является антагонистом LFA-1.
8. Модифицированный полипептид по п.7, отличающийся тем, что представляющий интерес полипептид является Fab или (Fab’)2 антитела против LFA-1.
9. Модифицированный полипептид по п.8, отличающийся тем, что представляющий интерес полипептид является анти-CD18 Fab или анти-CD18(Fab’)2.
10. Модифицированный полипептид по п.9, отличающийся тем, что представляющий интерес полипептид является человеческим или гуманизированным.
11. Модифицированный полипептид по любому одному из пп.1-10, отличающийся тем, что спасательный рецепторсвязывающий эпитоп содержит последовательность PKNSSMISNTR (SEQ ID NO:3).
12. Модифицированный полипептид по п.11, отличающийся тем, что спасательный рецепторсвязывающий эпитоп дополнительно содержит последовательность HQSLGTQ (SEQ ID NO:11).
13. Модифицированный полипептид по п.11, отличающийся тем, что спасательный рецепторсвязывающий эпитоп дополнительно содержит последовательность HQNLSDGK (SEQ ID NO:1).
14. Модифицированный полипептид по п.11, отличающийся тем, что спасательный рецепторсвязывающий эпитоп дополнительно содержит последовательность HQNISDGK (SEQ ID NO:2).
15. Модифицированный полипептид по п.11, отличающийся тем, что спасательный рецепторсвязывающий эпитоп дополнительно содержит последовательность VISSHLGQ (SEQ ID NO:31).
16. Модифицированный полипептид по п.1, отличающийся тем, что спасательный рецепторсвязывающий эпитоп содержит последовательности HQNLSDGK (SEQ ID NO:1), HQNISDGK (SEQ ID NO:2), HQSLGTQ (SEQ ID NO:11) или VISSHLGQ (SEQ ID NO:31) и последовательность PKNSSMISNTP (SEQ ID NO:3).
17. Модифицированный полипептид по п.16, отличающийся тем, что спасательный рецепторсвязывающий эпитоп слит с представляющим интерес полипептидом.
18. Молекула ДНК, кодирующая модифицированный полипептид по любому из пп.1-16.
19. Реплицируемый вектор, содержащий молекулу ДНК по п.18.
20. Способ получения модифицированного полипептида по любому из пп.1-17, включающий культивирование клеток, содержащих кодирующую указанный модифицированный полипептид нуклеиновую кислоту, в культуральной среде и извлечение указанного модифицированного полипептида из культуральной среды, клеток или как из культуральной среды, так и из клеток.
21. Способ получения модифицированного полипептида по любому из пп.1-15, включающий изменение представляющего интерес полипептида таким образом, что он содержит спасательный рецепторсвязывающий эпитоп Fc-области IgG и имеет увеличенное время полужизни in vivo.
22. Способ по п.21, отличающийся тем, что стадию изменения проводят с помощью сайт-направленного, кассетного или PCR-мутагенеза.
23. Способ получения модифицированного полипептида по п.1, включающий (а) идентификацию последовательности и конформации спасательного рецепторсвязывающего эпитопа Fc-области молекулы IgG, где указанный эпитоп происходит из СН2-домена Fc-области; б) изменение последовательности указанного представляющего интерес полипептида, который выводится через почки и не содержит Fc-область IgG, с целью включения последовательности и конформации идентифицированного связывающего эпитопа; (в) проверку измененного полипептида со стадии (б) на предмет более длительного времени полужизни in vivo, чем у представляющего интерес полипептида; (г) если измененный полипептид не обладает более длительным временем полужизни in vivo, дальнейшее изменение последовательности представляющего интерес полипептида с целью дальнейшего включения последовательности и конформации идентифицированного связывающего эпитопа и проверку на предмет увеличения времени полужизни in vivo до тех пор, пока не получат более длительное время полужизни in vivo.
24. Способ лечения опосредованного LFA-1 нарушения, характеризующийся тем, что млекопитающему, нуждающемуся в лечении, вводят эффективное количество модифицированного полипептида по любому из пп.1-15.
25. Способ лечения опосредованного LFA-1 нарушения, характеризующийся тем, что пациенту, нуждающемуся в лечении, вводят эффективное количество модифицированного полипептида по любому из пп.1-15.
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Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/422,093 | 1995-04-14 | ||
US08/422,093 US6096871A (en) | 1995-04-14 | 1995-04-14 | Polypeptides altered to contain an epitope from the Fc region of an IgG molecule for increased half-life |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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RU2003131869/13A Division RU2003131869A (ru) | 1995-04-14 | 2003-10-29 | Измененные полипептиды с увеличенным временем полужизни |
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RU97118592A RU97118592A (ru) | 1999-10-10 |
RU2224764C2 true RU2224764C2 (ru) | 2004-02-27 |
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RU97118592/13A RU2224764C2 (ru) | 1995-04-14 | 1996-03-28 | Модифицированный полипептид с увеличенным временем полужизни, его получение и использование |
RU2003131869/13A RU2003131869A (ru) | 1995-04-14 | 2003-10-29 | Измененные полипептиды с увеличенным временем полужизни |
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US (1) | US6096871A (ru) |
EP (1) | EP0821732B1 (ru) |
JP (2) | JPH11504509A (ru) |
CN (1) | CN100390279C (ru) |
AT (1) | ATE415474T1 (ru) |
AU (1) | AU721155B2 (ru) |
BR (1) | BR9604955A (ru) |
CA (1) | CA2217871C (ru) |
DE (1) | DE69637761D1 (ru) |
ES (1) | ES2317645T3 (ru) |
IL (1) | IL117850A0 (ru) |
MX (1) | MX9707877A (ru) |
NO (1) | NO326296B1 (ru) |
NZ (1) | NZ306170A (ru) |
RU (2) | RU2224764C2 (ru) |
WO (1) | WO1996032478A1 (ru) |
ZA (1) | ZA962489B (ru) |
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- 1996-03-28 AT AT96911483T patent/ATE415474T1/de not_active IP Right Cessation
- 1996-03-28 WO PCT/US1996/004316 patent/WO1996032478A1/en active Application Filing
- 1996-03-28 DE DE69637761T patent/DE69637761D1/de not_active Expired - Lifetime
- 1996-03-28 MX MX9707877A patent/MX9707877A/es active IP Right Grant
- 1996-03-28 BR BR9604955A patent/BR9604955A/pt not_active Application Discontinuation
- 1996-03-28 AU AU54359/96A patent/AU721155B2/en not_active Expired
- 1996-03-28 RU RU97118592/13A patent/RU2224764C2/ru active
- 1996-03-28 ZA ZA9602489A patent/ZA962489B/xx unknown
- 1996-03-28 CN CNB961943475A patent/CN100390279C/zh not_active Expired - Lifetime
- 1996-03-28 NZ NZ306170A patent/NZ306170A/en not_active IP Right Cessation
- 1996-03-28 EP EP96911483A patent/EP0821732B1/en not_active Expired - Lifetime
- 1996-03-28 ES ES96911483T patent/ES2317645T3/es not_active Expired - Lifetime
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2006
- 2006-11-30 JP JP2006324865A patent/JP4324610B2/ja not_active Expired - Fee Related
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Cited By (6)
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RU2517621C2 (ru) * | 2007-10-31 | 2014-05-27 | Ксенкор, Инк. | Fc-ВАРИАНТЫ С ИЗМЕНЕННЫМ СВЯЗЫВАНИЕМ С FcRn |
RU2529951C2 (ru) * | 2007-12-26 | 2014-10-10 | Ксенкор, Инк. | ВАРИАНТЫ Fc С ИЗМЕНЕННЫМ СВЯЗЫВАНИЕМ C FcRn |
RU2700882C2 (ru) * | 2007-12-26 | 2019-09-23 | Ксенкор, Инк. | ВАРИАНТЫ Fc С ИЗМЕНЕННЫМ СВЯЗЫВАНИЕМ С FcRn |
RU2696973C2 (ru) * | 2013-05-31 | 2019-08-07 | Ханми Фарм. Ко., Лтд. | FC-фрагмент IGG4, содержащий модифицированную шарнирную область |
RU2800558C1 (ru) * | 2013-05-31 | 2023-07-24 | Ханми Фарм. Ко., Лтд. | FC-фрагмент IgG4, содержащий модифицированную шарнирную область |
RU2714116C2 (ru) * | 2014-11-06 | 2020-02-11 | Ф. Хоффманн-Ля Рош Аг | ВАРИАНТЫ Fc-ОБЛАСТИ С МОДИФИЦИРОВАННЫМ СВЯЗЫВАНИЕМ FcRn И СПОСОБЫ ИХ ПРИМЕНЕНИЯ |
Also Published As
Publication number | Publication date |
---|---|
RU2003131869A (ru) | 2005-04-10 |
NO974739D0 (no) | 1997-10-13 |
EP0821732B1 (en) | 2008-11-26 |
ATE415474T1 (de) | 2008-12-15 |
AU5435996A (en) | 1996-10-30 |
NZ306170A (en) | 2001-06-29 |
AU721155B2 (en) | 2000-06-22 |
CA2217871C (en) | 2010-08-31 |
DE69637761D1 (de) | 2009-01-08 |
WO1996032478A1 (en) | 1996-10-17 |
ZA962489B (en) | 1997-09-29 |
CN1186517A (zh) | 1998-07-01 |
JP2007143552A (ja) | 2007-06-14 |
US6096871A (en) | 2000-08-01 |
NO326296B1 (no) | 2008-11-03 |
MX9707877A (es) | 1997-11-29 |
EP0821732A1 (en) | 1998-02-04 |
ES2317645T3 (es) | 2009-04-16 |
NO974739L (no) | 1997-12-12 |
BR9604955A (pt) | 1998-06-09 |
JP4324610B2 (ja) | 2009-09-02 |
IL117850A0 (en) | 1996-08-04 |
CA2217871A1 (en) | 1996-10-17 |
CN100390279C (zh) | 2008-05-28 |
JPH11504509A (ja) | 1999-04-27 |
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