RU2016138469A - Способ очистки конъюгатов на основе il-15/il-15r-альфа - Google Patents
Способ очистки конъюгатов на основе il-15/il-15r-альфа Download PDFInfo
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- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/20—Partition-, reverse-phase or hydrophobic interaction chromatography
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
- C07K14/7155—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for interleukins [IL]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K38/00—Medicinal preparations containing peptides
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Claims (19)
1. Способ получения композиции, содержащей мономерные конъюгаты, из образца, при этом указанный конъюгат содержит:
a) полипептид, содержащий аминокислотную последовательность интерлейкина 15 или его производных, и
b) полипептид, содержащий аминокислотную последовательность домена sushi IL-15Rα или его производных,
где указанный способ предусматривает применение анионообменной хроматографии с последующей хроматографией гидрофобного взаимодействия в отношении указанного образца.
2. Способ по п. 1, где указанный образец представляет собой образец среды для культивирования трансформированной клетки-хозяина, экспрессирующей конъюгат, при этом указанная клетка-хозяин представляет собой клетку млекопитающего, выбранную из группы, включающей клетки CHO, клетки HEK293, клетки COS, клетки PER.C6®, клетки SP2O, клетки NSO или любые клетки, происходящие от них, предпочтительно клетки CHO.
3. Способ по любому из пп. 1 или 2, где указанный способ включает стадии:
a) осуществления анионообменной хроматографии (AEX) в отношении образца с получением первого элюата и
b) осуществления хроматографии гидрофобного взаимодействия (HIC) в отношении первого элюата с получением второго элюата.
4. Способ по любому из пп. 1 или 2, где стадию a) анионообменной хроматографии (AEX) осуществляют с применением AEX-колонки или смолы, выбранной из группы, включающей Q Sepharose, например, Q Sepharose Fast Flow, Q Sepharose XL, Q Sepharose big beads, Q sepharose high performance, Q sepharose XL; DEAE Sephadex A-25, DEAE Sephadex A-50, QAE Sephadex A-25, QAE Sephadex A-50, Sourse 15Q, Sourse 30Q, Resourse Q, Capto Q, Capto DEAE, Mono Q, Toyopearl Super Q, Toyopearl DEAE, Toyopearl QAE, Toyopearl Q, Toyopearl GigaCap Q, TSKgel SuperQ, TSKgel DEAE, Fractogel EMD TMAE, Fractogel EMD TMAE HiCap, Fractogel EMD DEAE, Fractogel EMD DMAE, Macroprep High Q, Macro-prep-DEAE, Unosphere Q, Nuvia Q, POROS HQ, POROS PI, DEAE Ceramic HyperD, Q Ceramic HyperD, DEAE Sepharose Fast Flow и ANX Sepharose 4 Fast Flow.
5. Способ по п. 2, где на стадии a) осуществления анионообменной хроматографии (AEX) в отношении образца стадию загрузки проводят в буферном растворе с pH, равным 7,0 или более, предпочтительно pH, равным 7,5 или более.
6. Способ по любому из пп. 2 или 5, где на стадии b) осуществления хроматографии гидрофобного взаимодействия (HIC) в отношении образца стадию загрузки проводят в буферном растворе, содержащем соль аммония, предпочтительно сульфат аммония.
7. Способ по любому из пп. 2 или 5, где на стадии b) осуществления хроматографии гидрофобного взаимодействия (HIC) в отношении образца стадию загрузки осуществляют с помощью перемешивания первого элюата и буфера для загрузки непосредственно в ходе указанной хроматографии.
8. Способ по любому из пп. 2 или 5, где указанный способ дополнительно включает стадию применения полученного конъюгата для получения лекарственного препарата.
9. Фармацевтическая композиция, которая может быть получена с помощью способа по любому из пп. 1-8, при этом указанная композиция содержит по меньшей мере 80% мономерных конъюгатов.
10. Фармацевтическая композиция по п.9, где полипептиды a) и b) конъюгата ковалентно связаны в слитом белке.
11. Фармацевтическая композиция по любому из пп. 9 или 10, где указанный интерлейкин 15 имеет аминокислотную последовательность SEQ ID n°1, и домен sushi IL-15Rα имеет аминокислотную последовательность SEQ ID n°4.
12. Фармацевтическая композиция по любому из пп. 9 или 10, где указанный конъюгат содержит аминокислотную последовательность интерлейкина 15 или его производных в C-концевом положении по отношению к аминокислотной последовательности домена sushi IL-15Rα или его производных.
13. Фармацевтическая композиция по любому из пп. 9 или 10, где аминокислотная последовательность интерлейкина 15 или его производных и аминокислотная последовательность домена sushi IL-15Rα или производных разделены линкерной аминокислотной последовательностью с длиной 5-30 аминокислот, при этом указанный линкер содержит близкие к нейтральным аминокислоты, выбранные из группы, включающей Gly (G), Asn (N), Ser (S), Thr (T), Ala (A), Leu (L) и Gln (Q).
14. Композиция по любому из пп. 9 или 10 для лечения рака, инфекционного заболевания и/или иммунодефицитного нарушения у субъекта.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP14000742.8A EP2915569A1 (en) | 2014-03-03 | 2014-03-03 | IL-15/IL-15Ralpha based conjugates purification method |
EP14000742.8 | 2014-03-03 | ||
PCT/EP2015/000473 WO2015131994A1 (en) | 2014-03-03 | 2015-03-03 | Il-15/il-15ralpha based conjugates purification method |
Publications (3)
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RU2016138469A true RU2016138469A (ru) | 2018-04-04 |
RU2016138469A3 RU2016138469A3 (ru) | 2018-09-19 |
RU2701301C2 RU2701301C2 (ru) | 2019-09-25 |
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RU2016138469A RU2701301C2 (ru) | 2014-03-03 | 2015-03-03 | Способ очистки конъюгатов на основе il-15/il-15r-альфа |
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US (2) | US10808022B2 (ru) |
EP (2) | EP2915569A1 (ru) |
JP (2) | JP2017509618A (ru) |
KR (1) | KR102361954B1 (ru) |
CN (2) | CN106457070B (ru) |
AU (2) | AU2015226504B2 (ru) |
CA (1) | CA2938649C (ru) |
CY (1) | CY1123450T1 (ru) |
DK (1) | DK3113858T3 (ru) |
ES (1) | ES2807758T3 (ru) |
HR (1) | HRP20201122T1 (ru) |
HU (1) | HUE050138T2 (ru) |
IL (1) | IL247469B (ru) |
LT (1) | LT3113858T (ru) |
PL (1) | PL3113858T3 (ru) |
PT (1) | PT3113858T (ru) |
RS (1) | RS60627B1 (ru) |
RU (1) | RU2701301C2 (ru) |
SI (1) | SI3113858T1 (ru) |
WO (1) | WO2015131994A1 (ru) |
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