RU2377253C2 - Антитела, специфичные к фактору некроза опухолей, и их применение - Google Patents

Антитела, специфичные к фактору некроза опухолей, и их применение Download PDF

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RU2377253C2
RU2377253C2 RU2005116225/13A RU2005116225A RU2377253C2 RU 2377253 C2 RU2377253 C2 RU 2377253C2 RU 2005116225/13 A RU2005116225/13 A RU 2005116225/13A RU 2005116225 A RU2005116225 A RU 2005116225A RU 2377253 C2 RU2377253 C2 RU 2377253C2
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amino acid
acid sequence
monoclonal antibody
human monoclonal
light chain
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Джон С. БАБКУК (CA)
Джон С. БАБКУК
Джаспал С. КАНГ (CA)
Джаспал С. КАНГ
Орит ФУРД (US)
Орит ФУРД
Ларри ГРИН (US)
Ларри Грин
Ксиао ФЕНГ (US)
Ксиао Фенг
Скотт КЛАКАМП (US)
Скотт КЛАКАМП
Мэри ХААК-ФРЕНДСКО (US)
Мэри ХААК-ФРЕНДСКО
Паланисвами РАТАНАСВАМИ (CA)
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Крэйг ПИГОТ (CA)
Крэйг ПИГОТ
Мейна Л. ЛИАН (US)
Мейна Л. ЛИАН
Розан ЛИ (US)
Розан ЛИ
Кэти МАНЧУЛЕНЧО (CA)
Кэти МАНЧУЛЕНЧО
Рафаэлла ФАГИОНИ (US)
Рафаэлла ФАГИОНИ
Гиоргио СЕНАЛДИ (US)
Гиоргио СЕНАЛДИ
Киаожуан Джейн СУ (US)
Киаожуан Джейн СУ
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Амген Фремонт,Инк.
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Abstract

Изобретение относится к иммунологии и биотехнологии. Предложено моноклональное антитело человека, специфичное в отношении ФНО-альфа (TNF-alpha), содержащее легкую и тяжелую цепь с соответствующими CDR3 участками. Описаны его варианты, в том числе на основе тяжелой и легкой цепи, которые кодируются генами человека VH3-33 и A30VK1 или VH3-53 и L2VK3, соответственно. Раскрыты: способ оценки содержания ФНО-альфа (TNF-alpha) в образце пациента с использованием указанных антител и использование антител для приготовления лекарственного препарата. Описаны: составы для диагностики и лечения состояний, связанных с активностью ФНО-альфа (TNF-alpha) на основе антител. Раскрыта кодирующая нуклеиновая кислота, клетка для получения антител и способ получения антител. Использование изобретения обеспечивает высокоаффинные нейтрализующие моноклональные антитела, обладающие улучшенными Kd и IC50, по сравнению с Infliximab, Adalimumab или Etanercept, что может найти применение в медицине для лечения и диагностики заболеваний, связанных с повышенной активностью ФНО-альфа (TNF-alpha). 11 н. и 24 з.п. ф-лы, 13 ил., 36 табл.

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Claims (35)

1. Моноклональное антитело человека, которое специфическим образом связывается с фактором некроза опухолей альфа (ФНО-альфа) и содержит:
вариабельную область тяжелой цепи, которая содержит гипервариабельный участок 3 (CDR3) тяжелой цепи, который имеет последовательность аминокислот Glu Val Glu Ser Ala Met Gly Gly Phe Tyr Tyr Asn Gly Met Asp Val, и
вариабельную область легкой цепи, содержащую гипервариабельный участок 3 (CDR3) легкой цепи, который имеет последовательность аминокислот Leu Gin His Lys Ser Tyr Pro Leu Thr.
2. Моноклональное антитело человека по п.1, содержащее гипервариабельный участок 2 (CDR2) тяжелой цепи, который имеет последовательность аминокислот Val Ile Trp Ser Asp Gly Ser Ile Lys Tyr Tyr Ala Asp Ser Val Lys Gly.
3. Моноклональное антитело человека по п.1 или 2, содержащее гипервариабельный участок 1 (CDR1) тяжелой цепи, который имеет последовательность аминокислот Ser Tyr Asp Met His.
4. Моноклональное антитело человека по п.1 или 2, последовательность аминокислот тяжелой цепи которого содержит последовательность аминокислот SEQ ID NO: 70.
5. Моноклональное антитело человека по п.1 или 2, последовательность аминокислот тяжелой цепи которого содержит последовательность аминокислот SEQ ID NO: 74.
6. Моноклональное антитело человека по п.1 или 2, содержащее гипервариабельный участок 1 (CDR1) легкой цепи, который имеет последовательность аминокислот Arg Ala Ser Gln Gly lle Arg Ile Asp Leu Gly.
7. Моноклональное антитело человека по п.1 или 2, содержащее гипервариабельный участок 2 (CDR2) легкой цепи, который имеет последовательность аминокислот Ala Ala Ser Thr Leu Gln Ser.
8. Моноклональное антитело человека по п.1 или 2, последовательность аминокислот легкой цепи которого содержит последовательность аминокислот SEQ ID NO: 72.
9. Моноклональное антитело человека, которое специфически связывает фактор некроза опухолей альфа и содержит:
тяжелую цепь, которая содержит последовательность аминокислот, кодируемую геном VH3-33 человека, или ее консервативный вариант, и
легкую цепь, содержащую последовательность аминокислот, кодируемую геном легкой цепи A30VK1 человека, или ее консервативный вариант.
10. Моноклональное антитело человека, которое специфическим образом связывается с фактором некроза опухолей альфа и содержит:
вариабельную область тяжелой цепи, которая содержит гипервариабельный участок 3 (CDR3) тяжелой цепи, который имеет последовательность аминокислот Gly Glu Gly Gly Phe Asp Tyr, и
вариабельную область легкой цепи, которая содержит гипервариабельный участок 3 (CDR3) легкой цепи, который имеет последовательность аминокислот Gln Gln Tyr Asn Tyr Trp Trp Thr.
11. Моноклональное антитело человека по п.10, содержащее гипервариабельный участок 2 (CDR2) тяжелой цепи, который имеет последовательность аминокислот Val Ile Tyr Ser Gly Asp Arg Thr Tyr Tyr Ala Asp Ser Val Lys Gly.
12. Моноклональное антитело человека по п.10 или 11, содержащее гипервариабельный участок 1 (CDR1) тяжелой цепи, который имеет последовательность аминокислот Arg Asn Tyr Met Ser.
13. Моноклональное антитело человека по п.10 или 11, последовательность аминокислот тяжелой цепи которого содержит последовательность аминокислот SEQ ID NO: 50.
14. Моноклональное антитело человека по п.10 или 11, содержащее гипервариабельный участок 1 (CDR1) легкой цепи, который имеет последовательность аминокислот Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala.
15. Моноклональное антитело человека по п.10 или 11, содержащее гипервариабельный участок 2 (CDR2) легкой цепи, который имеет последовательность аминокислот Gly Ala Ser Ile Arg Ala Thr.
16. Моноклональное антитело человека по п.10 или 11, содержащее гипервариабельный участок 3 (CDR3) легкой цепи, который имеет последовательность аминокислот Gln Gln Tyr Asn Tyr Trp Trp Thr.
17. Моноклональное антитело человека по п.10 или 11, последовательность аминокислот легкой цепи которого содержит последовательность аминокислот SEQ ID NO: 52.
18. Моноклональное антитело человека, которое специфически связывает фактор некроза опухолей альфа и содержит:
тяжелую цепь, которая содержит последовательность аминокислот, кодируемую геном VH3-53 человека, или консервативный вариант указанной последовательности, и легкую цепь, которая содержит последовательность аминокислот, кодируемую геном легкой цепи L2VK3 человека, или консервативный вариант указанной последовательности.
19. Моноклональное антитело человека согласно любому из пп.1, 2, 9-11 и 18, которое связывает ФНОа с Kd менее 10-10 М.
20. Моноклональное антитело человека согласно п.19, которое связывает ФНОа с Kd менее 10-11 М.
21. Моноклональное антитело человека согласно любому из пп.1, 2, 9-11, 18, которое продуцировано линией гибридомы.
22. Моноклональное антитело человека согласно любому из пп.1, 2, 9-11, 18, которое продуцировано СНО клетками.
23. Моноклональное антитело человека согласно любому из пп.1, 2, 9-11, 18, которое представляет собой IgG1.
24. Моноклональное антитело человека согласно любому из пп.1, 2, 9-11, 18, которое представляет собой IgG2.
25. Способ оценки содержания фактора некроза опухолей альфа (ФНОα) в образце, взятом у пациента, согласно которому антитело против ФНОα согласно любому из пп.1-24 приводят в контакт с биологическим образцом, взятым у пациента, и определяют степень связывания указанного антитела с ФНОα в указанном образце, причем концентрацию ФНОα в указанном образце определяют на основании количества ФНОα, связанного с указанным антителом.
26. Способ по п.25, где биологический образец представляет собой кровь.
27. Состав для диагностики состояний, связанных с активностью и/или повышенной продукцией ФНОα, содержащий эффективное количество антитела по любому из пп.1-24 или его функциональный фрагмент и фармацевтически приемлемый носитель.
28. Применение антитела согласно любому из пп.1-24 для приготовления лекарственного препарата для эффективного лечения опухолевых заболеваний, иммунно-опосредованных воспалительных заболеваний или апоптоза, индуцированного фактором некроза опухолей у животного.
29. Применение по п.28, где указанное опухолевое заболевание выбирают из следующей группы: рак груди, рак яичников, рак желчного пузыря, рак легких, глиобластома, рак желудка, рак эндометрия, рак почек, рак толстой кишки, рак поджелудочной железы, рак простаты.
30. Применение по п.28, где указанное иммунно-опосредованное воспалительное заболевание выбирают из следующей группы: ревматоидный артрит, гламерулонефрит, атеросклероз, псориаз, рестеноз, аутоиммунная болезнь, болезнь Крона, отторжение трансплантата, септический шок, кахексия, анорексия, анкилозный спондилит, множественный склероз.
31. Применение по п.28, где указанное иммунно-опосредованное воспалительное заболевание выбирают из группы, включающей ревматоидный артрит, псориаз и болезнь Крона.
32. Нуклеиновая кислота, которая кодирует антитело согласно любому из пп.1-24.
33. Клетка-хозяин для продуцирования антител, охарактеризованных в любом из пп.1-24, которая содержит нуклеиновую кислоту по п.32.
34. Способ получения антитела, который включает
культивирование клетки по п.33 в условиях, при которых указанная нуклеиновая кислота экспрессируется с образованием антитела, и выделение антитела.
35. Состав для лечения состояний, связанных с активностью и/или повышенной продукцией ФНОα, содержащий эффективное количество антитела по любому из пп.1-24 или его функциональный фрагмент и фармацевтически приемлемый носитель.
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