RU2010145939A - Антигенсвязывающая молекула, способная к многократному связыванию двух или более молекул антигена - Google Patents
Антигенсвязывающая молекула, способная к многократному связыванию двух или более молекул антигена Download PDFInfo
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- 239000000427 antigen Substances 0.000 title claims abstract 168
- 108091007433 antigens Proteins 0.000 title claims abstract 168
- 102000036639 antigens Human genes 0.000 title claims abstract 168
- 230000027455 binding Effects 0.000 title claims abstract 148
- 238000000034 method Methods 0.000 claims abstract 40
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims abstract 22
- 230000000694 effects Effects 0.000 claims abstract 11
- 150000001413 amino acids Chemical class 0.000 claims abstract 10
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract 4
- 101710181478 Envelope glycoprotein GP350 Proteins 0.000 claims abstract 3
- 230000003042 antagnostic effect Effects 0.000 claims abstract 3
- 238000010494 dissociation reaction Methods 0.000 claims abstract 3
- 230000005593 dissociations Effects 0.000 claims abstract 3
- 230000003313 weakening effect Effects 0.000 claims abstract 3
- 238000012216 screening Methods 0.000 claims 15
- 108090000623 proteins and genes Proteins 0.000 claims 10
- 238000004519 manufacturing process Methods 0.000 claims 6
- 238000003780 insertion Methods 0.000 claims 1
- 230000037431 insertion Effects 0.000 claims 1
- 230000014759 maintenance of location Effects 0.000 claims 1
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Abstract
1. Антигенсвязывающая молекула, имеющая значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между KD для антигена при рН 5,8 и KD для антигена при рН 7,4, составляющее 2 или больше. ! 2. Антигенсвязывающая молекула по п.1, значение KD(pH 5,8)/KD(pH 7,4) которой составляет 10 или больше. ! 3. Антигенсвязывающая молекула по п.1, значение KD(pH 5,8)/KD(pH 7,4) которой составляет 40 или больше. ! 4. Антигенсвязывающая молекула по любому из пп.1-3, по крайней мере, одна аминокислота которой была замещена гистидином, или в которую был вставлен, по крайней мере, один гистидин. ! 5. Антигенсвязывающая молекула по любому из пп.1-3, которая обладает антагонистической активностью. ! 6. Антигенсвязывающая молекула по любому из пп.1-3, которая связывается с мембранным антигеном или растворимым антигеном. ! 7. Антигенсвязывающая молекула по любому из пп.1-3, которая является антителом. ! 8. Фармацевтическая композиция, включающая антигенсвязываюшую молекулу по любому из пп.1-7. ! 9. Способ улучшения фармакокинетики антигенсвязывающей молекулы путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4. ! 10. Способ увеличения числа раз связывания с антигеном для антигенсвязывающей молекулы путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4. ! 11. Способ увеличения числа антигенов, которое может быть связано антигенсвязывающей молекулой, путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4. ! 12. Способ диссоциации внутри клетки антигена от
Claims (50)
1. Антигенсвязывающая молекула, имеющая значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между KD для антигена при рН 5,8 и KD для антигена при рН 7,4, составляющее 2 или больше.
2. Антигенсвязывающая молекула по п.1, значение KD(pH 5,8)/KD(pH 7,4) которой составляет 10 или больше.
3. Антигенсвязывающая молекула по п.1, значение KD(pH 5,8)/KD(pH 7,4) которой составляет 40 или больше.
4. Антигенсвязывающая молекула по любому из пп.1-3, по крайней мере, одна аминокислота которой была замещена гистидином, или в которую был вставлен, по крайней мере, один гистидин.
5. Антигенсвязывающая молекула по любому из пп.1-3, которая обладает антагонистической активностью.
6. Антигенсвязывающая молекула по любому из пп.1-3, которая связывается с мембранным антигеном или растворимым антигеном.
7. Антигенсвязывающая молекула по любому из пп.1-3, которая является антителом.
8. Фармацевтическая композиция, включающая антигенсвязываюшую молекулу по любому из пп.1-7.
9. Способ улучшения фармакокинетики антигенсвязывающей молекулы путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
10. Способ увеличения числа раз связывания с антигеном для антигенсвязывающей молекулы путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
11. Способ увеличения числа антигенов, которое может быть связано антигенсвязывающей молекулой, путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
12. Способ диссоциации внутри клетки антигена от связавшейся с ним внеклеточно антигенсвязывающей молекулы путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
13. Способ высвобождения антигенсвязывающей молекулы, которая связалась с антигеном и подверглась интернализации в клетку, в свободной от антигена форме наружу из клетки путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
14. Способ увеличения способности антигенсвязывающей молекулы к элиминации антигена в плазме путем ослабления антигенсвязывающей активности антигенсвязывающей молекулы при рН 5,8 по сравнению с указанной активностью при рН 7,4.
15. Способ по любому из пп.9-14, в котором значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между KD для антигена при рН 5,8 и KD для антигена при рН 7,4, составляет 2 или больше.
16. Способ по любому из пп.9-14, в котором значение KD(pH 5,8)/KD(pH 7,4) составляет 10 или больше.
17. Способ по любому из пп.9-14, в котором значение KD(pH 5,8)/KD(pH 7,4) составляет 40 или больше.
18. Способ улучшения фармакокинетики антигенсвязывающей молекулы путем замены, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
19. Способ увеличения числа раз связывания с антигеном для антигенсвязывающей молекулы путем замены, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
20. Способ увеличения числа антигенов, которое может быть связано антигенсвязывающей молекулой, путем замены, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
21. Способ диссоциации внутри клетки антигена от связавшейся с ним внеклеточно антигенсвязывающей молекулы путем замены по крайней мере одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
22. Способ высвобождения антигенсвязывающей молекулы, которая связалась с антигеном и подверглась интернализации в клетку, в свободной от антигена форме наружу из клетки путем замены, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
23. Способ увеличения способности антигенсвязывающей молекулы к элиминации антигена в плазме путем замены, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
24. Способ по любому из пп.18-23, в котором замена гистидином или его вставка увеличивает значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между антигенсвязывающей активностью при рН 5,8 и антигенсвязывающей активностью при рН 7,4, по сравнению со значением KD(pH 5,8)/KD(pH 7,4) до замены гистидином или его вставки.
25. Способ по любому из пп.9-14 и 18-23, в котором антигенсвязывающая молекула обладает антагонистической активностью.
26. Способ по любому из пп.9-14 и 18-23, в котором антигенсвязывающая молекула связывается с мембранным антигеном или растворимым антигеном.
27. Способ по любому из пп.9-14 и 18-23, в котором антигенсвязывающей молекулой является антитело.
28. Способ скрининга антигенсвязывающей молекулы, который включает стадии:
(a) определения антигенсвязывающей активности антигенсвязывающей молекулы при рН от 6,7 до 10,0;
(b) определения антигенсвязывающей активности антигенсвязывающей молекулы при рН от 4,0 до 6,5; и
(c) отбора антигенсвязывающей молекулы, антигенсвязывающая активность которой при рН от 6,7 до 10,0 выше антигенсвязывающей активности при рН от 4,0 до 6,5.
29. Способ скрининга по п.28, который включает стадию отбора антитела, антигенсвязываюшая активность которого при рН от 6,7 до 10,0 - дважды или более антигенсвязывающей активности при рН от 4,0 до 6,5.
30. Способ скрининга антигенсвязывающей молекулы, который включает стадии:
(a) связывания антигенсвязывающей молекулы с антигеном при параметре рН, составляющем от 6,7 до 10,0;
(b) помещения антигенсвязывающей молекулы, которая связалась с антигеном на стадии (а), при параметре рН, составляющем от 4,0 до 6,5; и
(c) получения антигенсвязывающей молекулы, которая подвергалась диссоциации при параметре рН, составляющем от 4,0 до 6,5.
31. Способ скрининга для получения антигенсвязывающей молекулы, активность связывания которой при первом рН превышает активность связывания при втором рН, который включает стадии:
(a) связывания антигенсвязывающей молекулы с колонкой с иммобилизованным антигеном при параметре первого рН;
(b) элюирования антигенсвязывающей молекулы, которая связалась с колонкой при первом рН, с колонки при параметре второго рН; и
(c) сбора подвергшейся элюции антигенсвязывающей молекулы.
32. Способ скрининга для получения антигенсвязывающей молекулы, активность связывания которой при первом рН превышает активность связывания при втором рН, который включает стадии:
(а) связывания библиотеки антигенсвязывающих молекул с колонкой с иммобилизованным антигеном при параметре первого рН;
(b) элюирования антигенсвязывающей молекулы с колонки при параметре второго рН;
(c) амплификации гена, кодирующего элюируемую антигенсвязывающую молекулу; и
(d) получения элюируемой антигенсвязывающей молекулы.
33. Способ скрининга по п.31 или 32, в котором первый рН составляет от 6,7 до 10,0, а второй рН составляет от 4,0 до 6,5.
34. Способ скрининга по любому из пп.28-32, в котором, по крайней мере, одна или более аминокислот антигенсвязывающей молекулы была замещена гистидином, или, по крайней мере, один гистидин был вставлен в антигенсвязывающую молекулу.
35. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая обладает улучшенным сохранением в плазме.
36. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая способна к связыванию антигена два или более раз.
37. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая способна к связыванию большего числа антигенов, чем число ее антигенсвязывающих сайтов.
38. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая подвергает диссоциации связавшийся с ней внеклеточно антиген внутри клетки.
39. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая связывается с антигеном и подвергается интернализации в клетку, и высвобождается наружу из клетки в свободной от антигена форме.
40. Способ скрининга по любому из пп.28-32 для получения антигенсвязывающей молекулы, которая обладает увеличенной способностью к элиминации антигена в плазме.
41. Способ скрининга по любому из пп.28-32, в котором антигенсвязывающая молекула используется в виде фармацевтической композиции.
42. Способ скрининга по любому из пп.28-32, в котором антигенсвязывающей молекулой является антитело.
43. Способ продуцирования антигенсвязывающей молекулы, который включает стадии:
(a) определения антигенсвязывающей активности антигенсвязывающей молекулы при рН от 6,7 до 10,0;
(b) определения антигенсвязывающей активности антигенсвязывающей молекулы при рН от 4,0 до 6,5;
(c) отбора антигенсвязывающей молекулы, антигенсвязывающая активность которой при рН от 6,7 до 10,0 выше антигенсвязывающей активности при рН от 4,0 до рН 6,5;
(d) получения гена, кодирующего антигенсвязывающую молекулу, отобранную на стадии (с); и
(e) продуцирования антигенсвязывающей молекулы, используя полученный на стадии (d) ген.
44. Способ продуцирования антигенсвязывающей молекулы, который включает стадии:
(а) связывания антигенсвязывающей молекулы с антигеном при параметре рН, составляющем от 6,7 до 10,0;
(b) допущения нахождения антигенсвязывающей молекулы, связанной с антигеном на стадии (а), при параметре рН, составляющем от 4,0 до 6,5;
(c) сбора антигенсвязывающей молекулы, которая подвергалась диссоциации при параметре рН, составляющем от 4,0 до 6,5;
(d) получения гена, кодирующего антигенсвязывающую молекулу, полученную на стадии (с); и
(e) продуцирования антигенсвязывающей молекулы, используя полученный на стадии (d) ген.
45. Способ продуцирования антигенсвязывающей молекулы, активность связывания которой при первом рН превышает активность связывания при втором рН, который включает стадии:
(a) связывания антигенсвязывающей молекулы с колонкой с иммобилизованным антигеном при параметре первого рН;
(b) элюирования антигенсвязывающей молекулы, которая связалась с колонкой при первом рН, с колонки при параметре второго рН;
(c) сбора подвергшейся элюции антигенсвязывающей молекулы;
(d) получения гена, кодирующего антигенсвязывающую молекулу, полученную на стадии (с); и
(e) продуцирования антигенсвязывающей молекулы, используя полученный на стадии (d) ген.
46. Способ продуцирования антигенсвязывающей молекулы, активность связывания которой при первом рН превышает активность связывания при втором рН, который включает стадии:
(а) связывания библиотеки антигенсвязывающих молекул с колонкой с иммобилизованным антигеном при параметре первого рН;
(b) элюирования антигенсвязывающей молекулы с колонки при параметре второго рН;
(c) амплификации гена, кодирующего элюируемую антигенсвязывающую молекулу;
(d) сбора подвергшейся элюции антигенсвязывающей молекулы;
(e) получения гена, кодирующего антигенсвязывающую молекулу, собранную на стадии (d); и
(f) продуцирования антигенсвязывающей молекулы, используя полученный на стадии (е) ген.
47. Способ продуцирования по п.45 или 46, в котором первое рН составляет от 6,7 до 10,0, а второе рН составляет от 4,0 до 6,5.
48. Способ продуцирования по любому из пп.43-46, который дополнительно включает стадию замещения, по крайней мере, одной аминокислоты антигенсвязывающей молекулы гистидином или вставки, по крайней мере, одного гистидина в антигенсвязывающую молекулу.
49. Способ продуцирования по любому из пп.43-46, в котором антигенсвязывающей молекулой является антитело.
50. Фармацевтическая композиция, включающая антигенсвязывающую молекулу, полученную способом продуцирования по любому из пп.43-49.
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