KR20200006528A - 수의학 용도를 위한 il4/il13 수용체 분자 - Google Patents
수의학 용도를 위한 il4/il13 수용체 분자 Download PDFInfo
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- KR20200006528A KR20200006528A KR1020197032241A KR20197032241A KR20200006528A KR 20200006528 A KR20200006528 A KR 20200006528A KR 1020197032241 A KR1020197032241 A KR 1020197032241A KR 20197032241 A KR20197032241 A KR 20197032241A KR 20200006528 A KR20200006528 A KR 20200006528A
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Abstract
반려 동물 종으로부터 유래되고 IL13 및/또는 IL4에 결합하는 IL13R/IL4R 연속된 폴리펩타이드에 관한 다양한 구현예가 제공된다. 이러한 연속된 폴리펩타이드는 반려 동물, 예를 들면, 개과, 고양이과, 및 말과에서 IL13 및/또는 IL4-유도된 상태를 치료하는 방법에서 사용될 수 있다.
Description
관련 출원의 상호-참고
본 출원은 2018년 4월 21일자로 출원된 미국 가특허원 제62/488,509호의 우선권의 이익을 청구하며, 이는 임의의 목적을 위해 이의 전문이 본원에 참고로 포함된다.
분야
본 개시내용은 반려 동물 종의 IL4 및/또는 IL13, 예를 들면, 개과(canine) IL4 및 개과 IL13에 결합하는 반려 동물 종으로부터 유래된 인터루킨 4 수용체 및 인터루킨 13 수용체 단편을 포함하는 연속된(contiguous) 폴리펩타이드에 관한 것이다. 본 발명은 또한 세포, 예를 들면, 반려 동물, 예를 들면, 개과, 고양이과(feline) 및 말과(equine)에서 IL4 및/또는 IL13-유도된 상태를 치료하거나 IL4 및/또는 IL13 신호전달 활성을 감소시키기 위한, 연속된 폴리펩타이드의 사용 방법에 관한 것이다.
배경
인터루킨 4(IL4)는 천연의 헬퍼 T 세포의 Th2 세포로의 분화를 촉진하는 사이토킨이다. 인터루킨 13(IL13)은 면역 세포에서 유사한 효과를 갖는다. IL4 및 IL13 둘 다는 알레르기, 예를 들면, 아토피성 피부염 및 천식과 직접 관련된 T 세포-매개된 면역 반응에서 중요한 역활을 한다. IL4는 이종이량체성 수용체 IL4 수용체 소단위 알파(IL4R)와 γc 또는 IL4R 및 IL13 수용체 소단위 알파-1(IL13R)와 함께 신호전달 복합체를 형성할 수 있음이 일반적으로 이해되고 있다. IL13은 이종이량체성 수용체 IL4Ra 및 IL13Ra1과 신호전달 복합체를 형성할 수 있다. IL4Ra 또는 IL13Ra1의 세포외 도메인(extracellular domain)은 IL4 및/또는 IL13에 결합하여 사이토킨의 유리 농도를 감소시킬 수 있으므로, 피부염, 천식 및 기타 장애와 관련된 임상 신호 및 증상을 약화시킬 수 있다.
반려종 동물, 예를 들어, 고양이, 개, 및 말은 사람 알레르기성 질환, 예를 들면, 아토피성 피부염 및 천식과 유사한 많은 알레르기성 질환을 앓는다. 따라서, IL4/IL13-유도된 상태를 치료하고 IL4/IL13 신호전달 활성을 감소시키기 위해 반려 동물 IL4 및/또는 IL13에 결합시키는데 특이적으로 사용될 수 있는 방법 및 화합물에 대한 요구가 남아있다.
요약
일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인 및 IL4R 폴리펩타이드의 세포외 도메인을 포함하는 IL13R/IL4R 연속된 폴리펩타이드가 제공되며, 여기서 IL13R 및 IL4R 폴리펩타이드는 반려 동물 종으로부터 유래된다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 식 (I) IL13R-L1-IL4R-L2-FP 또는 식 (II) IL4R-L1-IL13R-L2-FP를 포함하며, 여기서:
a. IL13R은 반려 동물 종으로부터 유래된 IL13R 폴리펩타이드의 세포외 도메인이고,
b. IL4R는 반려 동물 종으로부터 유래된 IL4R 폴리펩타이드의 세포외 도메인이며,
c. L1은 제1의 임의의 링커이고,
d. L2는 제2의 임의의 링커이며,
e. FP는 융합 파트너이다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 생물층 간섭법(biolayer interferometry)에 의해 측정된 바와 같이, 반려 동물 종의 IL13에 5 x 10-6 M 미만, 1 x 10-6 M 미만, 5 x 10-7 M 미만, 1 x 10-7 M 미만, 5 x 10-8 M 미만, 1 x 10-8 M 미만, 5 x 10-9 M 미만, 1 x 10-9 M 미만, 5 x 10-10 M 미만, 1 x 10-10 M 미만, 5 x 10-11 M 미만, 1 x 10-11 M 미만, 5 x 10-12 M 미만, 또는 1 x 10-12 M 미만의 해리 상수(Kd)로 결합한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 생물층 간섭법에 의해 측정된 바와 같이, 반려 동물 종의 IL4에 5 x 10-6 M 미만, 1 x 10-6 M 미만, 5 x 10-7 M 미만, 1 x 10-7 M 미만, 5 x 10-8 M 미만, 1 x 10-8 M 미만, 5 x 10-9 M 미만, 1 x 10-9 M 미만, 5 x 10-10 M 미만, 1 x 10-10 M 미만, 5 x 10-11 M 미만, 1 x 10-11 M 미만, 5 x 10-12 M 미만, 또는 1 x 10-12 M 미만의 해리 상수로 결합한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 반려 동물 종에서 IL13 및/또는 IL4 신호전달을 감소시킨다.
일부 구현예에서, 반려 동물 종은 개과, 고양이과, 또는 말과이다.
일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 85% 동일하다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 90% 동일하다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 95% 동일하다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 98% 동일하다.
일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22의 18번 위치에 상응하거나, 서열 번호: 24의 18번 위치에 상응하거나, 서열 번호: 26의 18번 위치에 상응하는 위치에서 시스테인을 포함한다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22의 18번 위치에서, 서열 번호: 24의 18번 위치에서, 서열 번호: 26의 18번 위치에서, 서열 번호: 32의 15번 위치에서, 서열 번호: 34의 15번 위치에서, 또는 서열 번호: 36의 15번 위치에서 시스테인을 포함한다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 32, 서열 번호: 34, 및 서열 번호: 36으로부터 선택된 아미노산 서열을 포함한다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 및 서열 번호: 26으로부터 선택된 아미노산 서열을 포함한다.
일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 85% 동일하다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 90% 동일하다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산에 대해 적어도 95% 동일하다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 98% 동일하다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 33, 서열 번호: 35, 및 서열 번호: 37로부터 선택된 아미노산 서열을 포함한다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 및 서열 번호: 27로부터 선택된 아미노산 서열을 포함한다.
일부 구현예에서, L1 및 L2는, 존재하는 경우, 각각 독립적으로 G, GG, GGG, S, SS, SSS, GS, GSGS(서열 번호: 38), GSGSGS(서열 번호: 39), GGS, GGSGGS(서열 번호: 40), GGSGGSGGS(서열 번호: 41), GGGS(서열 번호: 42), GGGSGGGS(서열 번호: 43), GGGSGGGSGGGS(서열 번호: 44), GSS, GSSGSS(서열 번호: 45), GSSGSSGSS(서열 번호: 46), GGSS(서열 번호: 47), GGSSGGSS(서열 번호: 48), 및 GGSSGGSSGGSS(서열 번호: 49)로부터 선택된 아미노산 서열을 포함한다.
일부 구현예에서, 융합 파트너는 Fc, 알부민, 및 알부민 결합 단편으로부터 선택된다. 일부 구현예에서, Fc는 (a) IgG-A, IgG-B, IgG-C, 및 IgG-D 불변 영역으로부터 선택된 개과 중쇄 불변 영역; (b) IgG1, IgG2a, 및 IgG2b 불변 영역으로부터 선택된 고양이과 중쇄 불변 영역; 또는 (c) IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 및 IgG7 불변 영역으로부터 선택된 말과 중쇄 불변 영역이다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 13, 서열 번호: 14, 서열 번호: 15, 서열 번호: 16, 서열 번호: 17, 서열 번호 18, 서열 번호: 19, 서열 번호: 20, 서열 번호: 21, 서열 번호: 28, 서열 번호: 29, 서열 번호: 30, 및 서열 번호: 31로부터 선택된 서열을 포함한다.
일부 구현예에서, 상기 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드를 암호화하는, 단리된 핵산이 제공된다. 일부 구현예에서, 상기 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드를 암호화하는 핵산을 포함하는 숙주 세포가 제공된다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드를 생산하는 방법이 제공되며, 이는 상기 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드를 암호화하는 핵산을 포함하는 이러한 숙주 세포를 배양하는 단계 및 연속된 폴리펩타이드를 단리하는 단계를 포함한다. 일부 구현예에서, 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드 및 약제학적으로 허용되는 담체를 포함하는 약제학적 조성물이 제공된다.
일부 구현예에서, 반려 동물 종에게 치료학적 유효량의 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드 또는 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물을 투여하는 단계를 포함하는, IL13 및/또는 IL4-유도된 상태를 갖는 반려 동물 종을 치료하는 방법이 제공된다. 일부 구현예에서, 반려 동물 종은 개과, 고양이과, 또는 말과이다. 일부 구현예에서, IL13 및/또는 IL4-유도된 상태는 소양증 또는 알레르기 상태, 예를 들면, 아토피성 피부염, 소양증, 천식, 건선, 피부경화증, 또는 습진이다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물은 비경구적으로 투여된다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물은 근육내 경로, 복강내 경로, 뇌척수내 경로, 피하 경로, 동맥내 경로, 활액내 경로, 수막내 경로, 또는 흡입 경로에 의해 투여된다.
일부 구현예에서, 이러한 방법은 Jak 억제제, PI3K 억제제, AKT 억제제, 또는 MAPK 억제제를 투여함을 추가로 포함한다. 일부 구현예에서, 이러한 방법은 항-IL17 항체, 항-IL31 항체, 항-TNFα 항체, 항-CD20 항체, 항-CD19 항체, 항-CD25 항체, 항-IL4 항체, 항-IL13 항체, 항-IL23 항체, 항-IgE 항체, 항-CD11α 항체, 항-IL6R 항체, 항-α4-인테그린 항체, 항-IL12 항체, 항-IL1β 항체, 및 항-BlyS 항체로부터 선택된 하나 이상의 항체를 투여함을 추가로 포함한다.
일부 구현예에서, 세포를 IL13 및/또는 IL4에 대한 연속된 폴리펩타이드의 결합을 허용하는 조건하에서 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 노출시킴으로써, (a) 천연의 IL13 수용체 및/또는 천연의 IL-4 수용체에 대한 IL/4 및/또는 IL-13의 결합을 감소시키고 IL13- 및/또는 IL-4-매개된 신호전달을 감소시킴을 포함하는, 세포내에서 IL13 및/또는 IL4 신호전달 활성을 감소시키는 방법이 제공된다. 일부 구현예에서, 세포는 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 생체외(ex vivo)에서 노출된다. 일부 구현예에서, 세포는 연속된 폴리펩타이드 또는 약제학적 조성물에 생체내에서 노출된다. 일부 구현예에서, 세포는 개과 세포, 고양이과 세포, 또는 말과 세포이다.
일부 구현예에서, 샘플을 IL13 및/또는 IL4에 대한 연속된 폴리펩타이드의 결합을 허용하는 조건 하에서 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물과 접촉시키고, 복합체가 샘플 속의 연속된 폴리펩타이드와 IL13 및/또는 IL4 사이에 형성되는지를 검출함을 포함하여, 반려 동물 종으로부터의 샘플 속에서 IL13 또는 IL4를 검출하는 방법이 제공된다. 일부 구현예에서, 샘플은 개과, 고양이과, 또는 말과로부터 수득된 생물학적 샘플이다.
도 1은 PBS 중 30μg/mL의 IL4 및 IL13의 농도를 사용하여 개과 IL4 및 IL13 또는 개과 IL13 및 IL4에 대한 개과 IL4R-IL13R-Fc 순차적인 결합의 그래프이다.
도 2는 PBS 중 30μg/mL of IL4 및 IL13의 농도를 사용하여 개과 IL4 및 IL13 또는 개과 IL13 및 IL4에 대한 개과 IL13R-IL4R-Fc 순차적 결합의 그래프이다.
도 3은 TF1 세포 증식 검정에서 개과 IL4R-IL13R-Fc 중화 개과 IL4 활성의 그래프이다. 개과 IL4(50 ng/mL 또는 3.85 nM)가 검정에 사용되었다.
도 2는 PBS 중 30μg/mL of IL4 및 IL13의 농도를 사용하여 개과 IL4 및 IL13 또는 개과 IL13 및 IL4에 대한 개과 IL13R-IL4R-Fc 순차적 결합의 그래프이다.
도 3은 TF1 세포 증식 검정에서 개과 IL4R-IL13R-Fc 중화 개과 IL4 활성의 그래프이다. 개과 IL4(50 ng/mL 또는 3.85 nM)가 검정에 사용되었다.
서열의 설명
표 1은 본원에 언급된 특정 서열의 목록을 제공한다.
구현예의 설명
개과 IL13 및/또는 IL4, 고양이과 IL13 및/또는 IL4, 및/또는 말과 IL13 및/또는 IL4에 결합하는 연속된 폴리펩타이드가 제공된다. 일부 구현예에서, 연속된 폴리펩타이드는 IL13R 폴리펩타이드의 세포외 도메인 및 IL4R 폴리펩타이드의 세포외 도메인을 포함한다. 연속된 폴리펩타이드를 생산 또는 정제하는 방법이 또한 제공된다. 연속된 폴리펩타이드를 사용하여 IL13 및/또는 IL4에 결합시켜 IL13- 및/또는 IL-4-매개된 신호전달을 억제하는 처리 방법이 제공된다. 이러한 방법은 반려 동물 종에서 IL13- 및/또는 IL4-유도된 상태를 치료하는 방법을 포함하지만, 이에 한정되지 않는다. 반려 동물 종으로부터의 샘플 속에서 IL13 및/또는 IL4를 검출하는 방법이 또한 제공된다.
독자의 편의를 위해, 본원에 사용된 용어의 다음의 정의를 제공한다.
본원에 사용된 바와 같이, Kd와 같은 수치 용어는 과학적 측정을 기반으로 계산되므로, 적절한 측정 오차에 속한다. 일부 예에서, 수치 용어는 가장 근접하게 유의적인 수치를 아우르는 수치 값을 포함할 수 있다.
본원에 사용된 바와 같이, 단수("a" 또는 "an")는 달리 명시하지 않는 한 "적어도 하나" 또는 "하나 이상"을 의미한다. 본원에 사용된 바와 같이, 용어 "또는"은 달리 명시하지 않는 한 "및/또는"을 의미한다. 다수의 종속항의 맥락에서, 다른 청구항들을 다시 언급하는 경우 "또는"의 사용은 이들 청구항을 대안적으로만 지칭한다.
IL13R/IL4R 연속된 폴리펩타이드
신규한 IL13R/IL4R 연속된 폴리펩타이드, 예를 들면, 개과 IL13 및/또는 IL4, 고양이과 IL13 및/또는 IL4, 및/또는 말과 IL13 및/또는 IL4에 결합하는 연속된 폴리펩타이드가 제공된다.
"아미노산 서열"은 펩타이드 또는 단백질내 아미노산 잔기의 서열을 의미한다. 용어 "폴리펩타이드" 및 "단백질"은 상호교환적으로 사용되어 아미노산 잔기의 중합체를 지칭하며, 최소 길이에 한정되지 않는다. 아미노산 잔기의 이러한 중합체는 천연 또는 비-천연 아미노산 잔기를 함유할 수 있고, 아미노산 잔기의 펩타이드, 올리고펩타이드, 이량체, 삼량체, 및 다량체를 포함하나, 이에 한정되지 않는다. 완전한 길이의 단백질 및 이의 단편 둘 다는 이러한 정의에 포함된다. 이러한 용어는 또한 폴리펩타이드의 발현 후 변형(post-expression modification), 예를 들면, 글리코실화, 시알릴화, 아세틸화, 포스포릴화 등을 포함한다. 또한, 본 개시내용의 목적을 위하여, "폴리펩타이드"는 단백질이 목적한 활성을 유지하는 한, 천연 서열에 대한 변형, 예를 들면, 결실, 첨가, 및 치환(일반적으로 천연에서 보존적인)을 포함하는 단백질을 지칭한다. 이러한 변형은 부위-지시된 돌연변이유발을 통해서와 같이 의도적일 수 있거나, 단백질을 생산하는 숙주의 돌연변이 또는 PCR 증폭으로 인한 오차를 통하는 것과 같이 우연일 수 있다.
본원에서 용어 "연속된 폴리펩타이드"는 아미노산의 차단되지 않은 서열을 의미하는데 사용된다. 연속된 폴리펩타이드는 전형적으로 단일의 연속 DNA 서열로부터 해독된다. 이는 유전 가공, 예를 들면, 제1 단백질의 DNA 서열로부터 정지 코돈을 제거한 후, 골격내(in frame)에 제2 단백질의 DNA 서열을 첨부함으로써, DNA 서열이 단일 단백질로서 발현되도록 하여 이룰 수 있다. 전형적으로, 이는 cDNA를 기존의 유전자와 골격내에서 발현 벡터내로 클로닝함으로써 달성된다.
본원에 사용된 바와 같이, "IL4R"은 IL-4에 결합하는 IL4 수용체 소단위 알파의 전체 또는 소 단위를 포함하는 폴리펩타이드이다.
예를 들면, "IL4R"은 달리 나타내지 않는 한, 영장류(예컨대, 사람 및 시노몰구스 원숭이(cynomolgus monkey)), 설치류(예컨대, 마우스 및 랫트), 및 반려 동물(예컨대, 개, 고양이, 및 말)과 같은 포유동물을 포함하는 임의의 척추동물원으로부터의 IL4R 폴리펩타이드를 지칭한다. 일부 구현예에서, IL4R는 IL4에 결합하는 세포외 도메인 단편이다. 일부 이러한 구현예에서, IL4R는 IL4R 세포외 도메인(ECD)으로 지칭될 수 있다. 일부 구현예에서, IL4R은 서열 번호: 7, 서열 번호:8, 서열 번호: 9, 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열을 포함한다.
본원에 사용된 바와 같이, "IL13R"은 IL-13에 결합하는 IL13 수용체 소단위 알파-1의 전체 또는 일부를 포함하는 폴리펩타이드이다.
예를 들면, "IL13R"은 달리 나타내지 않는 한, 영장류(예컨대, 사람 및 시노몰구스 원숭이), 설치류(예컨대, 마우스 및 랫트), 및 반려 동물(예컨대, 개, 고양이, 및 말)과 같은 포유동물을 포함하는 임의의 척추동물원으로부터의 IL13R 폴리펩타이드를 지칭한다. 일부 구현예에서, IL13R은 IL13에 결합하는 세포외 도메인 단편이다. 일부 이러한 구현예에서, IL13R은 IL13R 세포외 도메인(ECD)으로서 지칭될 수 있다. 일부 구현예에서, IL13R 폴리펩타이드는 서열 번호: 10, 서열 번호: 11, 서열 번호: 12, 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열을 포함한다.
용어 "반려 동물 종"은 사람에게 반려가 되기에 적합한 동물을 지칭한다. 일부 구현예에서, 반려 동물 종은 작은 포유동물, 예를 들면, 개과, 고양이과, 개, 고양이, 말, 토끼, 흰담비, 기니아 피그(guinea pig), 설치류 등이다. 일부 구현예에서, 반려 동물 종은 농장 동물, 예를 들면, 말, 소, 돼지 등이다.
"세포외 도메인"("ECD")은 세포외 공간내로 막관통 도메인(transmembrane domain)을 능가하여 연장하는 폴리펩타이드의 부위이다. 본원에 사용된 바와 같이, 용어 "세포외 도메인"은 완전한 세포외 도메인을 포함할 수 있거나 이의 리간드에 결합하는 하나 이상의 아미노산을 결여한 절단된(truncated) 세포외 도메인을 포함할 수 있다. 세포외 도메인의 조성물은 어느 아미노산이 막 내에 존재하는지를 결정하는데 사용된 알고리즘에 의존할 수 있다. 상이한 알고리즘이 예측될 수 있으며, 상이한 시스템은 제공된 단백질에 대한 상이한 세포외 도메인을 발현할 수 있다.
IL4R 폴리펩타이드의 세포외 도메인은 IL4에 결합하는 IL4R의 완전한 세포외 도메인 또는 절단된 세포외 도메인을 포함할 수 있다. 본원에 사용된 바와 같이, 용어 "IL4R 폴리펩타이드의 세포외 도메인", "IL4R ECD" 및 유사 용어는 심지어 이러한 용어가 "포함하는", "포함하다" 등과 같은 개방된 변천 단어를 따르는 경우에도, 막횡단 도메인 또는 세포질성 도메인을 포함하지 않는 IL4R 폴리펩타이드를 지칭한다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 반려 종 동물로부터 유래된 IL4R 폴리펩타이드의 세포외 도메인이다. 예를 들면, 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 개과 IL4R, 고양이과 IL4R 또는 말과 IL4R로부터 유래된다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 23, 서열 번호: 25, 또는 서열 번호: 27의 아미노산 서열, 또는 이의 임의의 단편을 포함한다. 일부 구현예에서, IL4R 폴리펩타이드의 세포외 도메인은 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열, 또는 이의 임의의 단편을 포함한다.
IL13R 폴리펩타이드의 세포외 도메인은 IL13에 결합하는 IL13R의 완전한 세포외 도메인 또는 절단된 세포외 도메인을 포함할 수 있다. 본원에 사용된 바와 같이, 용어 "IL13R 폴리펩타이드의 세포외 도메인", "IL13R ECD" 및 유사 용어는 심지어 이러한 용어가 "포함하는", "포함하다" 등과 같은 개방된 변천 단어를 따르는 경우에도, 막횡단 도메인 또는 세포질성 도메인을 포함하지 않는 IL13R 폴리펩타이드를 지칭한다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 반려 종 동물로부터 유래된 IL13R 폴리펩타이드의 세포외 도메인이다. 예를 들면, 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 개과 IL13R, 고양이과 IL13R 또는 말과 IL13R로부터 유래된다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 22, 서열 번호: 24, 또는 서열 번호: 26의 아미노산 서열, 또는 이의 임의의 단편을 포함한다. 일부 구현예에서, IL13R 폴리펩타이드의 세포외 도메인은 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열, 또는 이의 임의의 단편을 포함한다.
용어 "IL13R/IL4R 연속된 폴리펩타이드" 및 "IL4R/IL13R 연속된 폴리펩타이드"는 상호교환적으로 사용되어 IL13R 폴리펩타이드 및 IL4R 폴리펩타이드를 포함하는 연속된 폴리펩타이드를 지칭하고, 여기서 이러한 용어는 순서를 나타내지 않으며 여기서 IL13R 및 IL4R 폴리펩타이드는 순서를 달리 나타내지 않는 한, 연속된 폴리펩타이드를 나타낸다. 예를 들면, IL13R/IL4R 연속된 폴리펩타이드 또는 IL4R/IL13R 연속된 폴리펩타이드는 IL13R 폴리펩타이드가 서열의 앞 또는 뒤에서 오는 IL4R 폴리펩타이드를 지칭할 수 있다. 또한, IL13R/IL4R 연속된 폴리펩타이드 또는 IL4R/IL13R 연속된 폴리펩타이드는 IL4R 폴리펩타이드가 서열의 앞 또는 뒤에서 오는 IL13R 폴리펩타이드를 지칭할 수 있다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 IL13R 폴리펩타이드의 C-말단 또는 IL13R 폴리펩타이드의 N-말단에서 IL4R 폴리펩타이드에 결합된 IL13R 폴리펩타이드를 포함한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 IL4R 폴리펩타이드의 C-말단 또는 IL4R 폴리펩타이드의 N-말단에서 IL13R 폴리펩타이드에 결합된 IL4R 폴리펩타이드를 포함한다.
본 발명의 IL13R/IL4R 연속된 폴리펩타이드는 IL13R 폴리펩타이드의 세포외 도메인 및/또는 IL4R 폴리펩타이드의 세포외 도메인을 포함할 수 있으며, 여기서 폴리펩타이드는 반려 동물 종으로부터 유래된다. 예를 들면, 연속된 폴리펩타이드는 개, 고양이, 또는 말로부터의 IL4R 폴리펩타이드의 세포외 도메인을 포함할 수 있고/있거나 개, 고양이, 또는 말로부터의 IL13R 폴리펩타이드의 세포외 도메인을 포함할 수 있다.
"야생형"은 천연적으로 존재하는 폴리펩타이드의 돌연변이되지 않는 버젼, 또는 이의 단편을 지칭한다. 야생형 폴리펩타이드는 재조합적으로 생산될 수 있다. "야생형 IL13R ECD" 또는 "야생형 IL4R ECD"는 천연에서 발생하는 IL13R 또는 IL4R의 세포외 도메인의 동일한 부위와 동일한 아미노산 서열을 갖는 단백질을 지칭한다.
"변이체"는 단일 또는 다수의 아미노산 치환, 결실 및/또는 첨가에 의해 참고 핵산 분자 또는 폴리펩타이드로부터 상이하고 참고 핵산 분자 또는 폴리펩타이드의 적어도 하나의 생물학적 활성을 실질적으로 보유하는 핵산 분자 또는 폴리펩타이드이다.
"생물학적으로 활성인" 실체, 또는 "생물학적 활성"을 갖는 실체는 물질대사 또는 생리학적 공정과 관련되거나 연관된 임의의 기능을 가지고/가지거나 천연적으로 존재하는 분자의 구조적, 조절성, 또는 생화학적 기능을 갖는 실체이다. 생물학적으로 활성인 폴리뉴클레오타이드 단편은 본 발명의 폴리뉴클레오타이드의 활성에 대해 유사한 활성을 나타내지만, 필수적으로 동일하지 않는 것들이다. 생물학적으로 활성인 폴리펩타이드 또는 이의 단편은 생물학적 반응에 관여할 수 있는 것, 예를 들면, 그러나 이에 한정되지 않는 리간드-수용체 상호작용 또는 항원-항체 결합을 포함한다. 생물학적 활성은 개선된 바람직한 활성, 또는 감소된 바람직하지 않는 활성을 포함할 수 있다. 실체는 이것이 다른 분자와의 분자 상호작용, 예를 들면, 하이브리드화에 관여하는 경우, 이것이 질환 상태를 완화시키는 치료학적 가치를 가지는 경우, 이것이 면역 반응을 유도하는 예방학적 가치를 가지는 경우, 이것이 분자, 예를 들면, 폴리뉴클레오타이드 분자에 대해 유일한 것으로 측정될 수 있는 폴리뉴클레오타이드의 생물학적으로 활성인 단편의 존재를 측정하는데 있어서 진단학적 및/또는 예후적 가치를 갖는 경우, 및 이것이 폴리머라제 연쇄 반응(PCR)에서 프라이머(primer)로서 사용될 수 있는 경우, 생물학적 활성을 입증할 수 있다.
본원에 사용된 바와 같이, 핵산 분자 또는 폴리펩타이드 서열과 관련하여 "아미노산 서열 동일성 퍼센트(%)" 및 "상동성"은 서열을 정렬하여 갭(gap)을 도입한 후, 필요한 경우, 최대의 서열 동일성 퍼센트를 달성하고, 서열 동일성의 일부로서 임의의 보존적 치환을 고려하지 않는 경우, 특정 핵산 분자 또는 폴리펩타이드 서열내 뉴클레오타이드 또는 아미노산 잔기와 동일한 참고 서열내 뉴클레오타이드 또는 아미노산 잔기의 퍼센트로서 정의된다. 서열 동일성 퍼센트를 측정할 목적의 정렬은 당해 분야의 기술내에 있는 다양한 방법, 예를 들면, BLAST, BLAST-2, ALIGN, 또는 MEGALINETM(DNASTAR) 소프트웨어와 같은 공공 이용가능한 컴퓨터 소프트웨어를 사용하여 달성할 수 있다. 당해 분야의 기술자는 비교하는 서열의 전체 길이에 걸쳐 최대의 정렬을 달성하는데 요구되는 임의의 알고리즘을 포함하는, 정렬을 측정하기 위한 적절한 매개변수를 측정할 수 있다.
일부 구현예에서, 변이체는 서열을 정렬하고 갭을 도입한 후, 필요한 경우, 최대 서열 동일성 퍼센트를 달성하고, 서열 동일성의 일부로서 임의의 보존적 치환을 고려하지 않는 참고 핵산 분자 또는 폴리펩타이드와 적어도 약 50%의 서열 동일성을 갖는다. 이러한 변이체는 예를 들면, 폴리펩타이드의 N- 또는 C-말단에서 하나 이상의 아미노산 잔기가 첨가되거나 결실된 폴리펩타이드를 포함한다. 일부 구현예에서, 변이체는 참고 핵산 또는 폴리펩타이드의 서열과 적어도 약 50% 서열 동일성, 적어도 약 60% 서열 동일성, 적어도 약 65% 서열 동일성, 적어도 약 70% 서열 동일성, 적어도 약 75% 서열 동일성, 적어도 약 80% 서열 동일성, 적어도 약 85% 서열 동일성, 적어도 약 90% 서열 동일성, 적어도 약 95% 서열 동일성, 적어도 약 98% 서열 동일성을 갖는다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 85%, 적어도 90%, 적어도 95%, 적어도 98% 서열 동일성을 갖는 IL13R 폴리펩타이드의 세포외 도메인을 포함한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 85%, 적어도 90%, 적어도 95%, 적어도 98% 서열 동일성을 갖는 IL4R 폴리펩타이드의 세포외 도메인을 포함한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 22의 18번 위치에 상응하는 위치, 서열 번호: 24의 18번 위치에 상응하는 위치, 또는 서열 번호: 26의 의 18번 위치에 상응하는 위치에 시스테인을 포함하는 IL13R 폴리펩타이드의 세포외 도메인을 포함한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 22의 18번 위치에서, 서열 번호: 24의 18번 위치에서, 서열 번호: 26의 18번 위치에서, 서열 번호: 32의 15번 위치에서, 서열 번호: 34의 15번 위치에서, 또는 서열 번호: 36의 15번 위치에서 시스테인을 포함하는 IL13R 폴리펩타이드의 세포외 도메인을 포함한다.
"점 돌연변이(point mutation)"는 단일 뉴클레오타이드 또는 아미노산 잔기를 포괄하는 돌연변이이다. 이러한 돌연변이는 뉴클레오타이드 또는 아미노산의 손실, 하나의 뉴클레오타이드 또는 아미노산 잔기의 다른 것으로의 치환, 또는 추가의 뉴클레오타이드 또는 아미노산 잔기의 삽입일 수 있다.
아미노산 치환은 폴리펩타이드내 하나의 아미노산의 다른 아미노산으로의 대체를 포함할 수 있으나 이에 한정되지 않는다. 예시적인 치환은 표 2에 나타낸다. 아미노산 치환은 목적한 활성, 예를 들면, 보유된/개선된 항원 결합, 감소된 면역원성, 또는 개선된 ADCC 또는 CDC 또는 향상된 약동학에 대해 스크리닝된 목적한 분자 및 생성물내로 도입될 수 있다.
아미노산은 일반적인 측쇄 특성에 따라 그룹화될 수 있다:
(1) 소수성: 노르루이신, Met, Ala, Val, Leu, Ile;
(2) 중성 친수성: Cys, Ser, Thr, Asn, Gln;
(3) 산성: Asp, Glu;
(4) 염기성: His, Lys, Arg;
(5) 쇄 배향에 영향을 미치는 잔기: Gly, Pro;
(6) 방향족: Trp, Tyr, Phe.
비-보존적 치환은 이러한 부류 중 하나의 구성원을 다른 부류로 교환함을 포함할 것이다.
본원에 사용된 바와 같이, "융합 파트너"는 IL13R/IL4R 연속된 폴리펩타이드의 추가의 성분, 예를 들면, 추가의 폴리펩타이드, 예를 들면, 알부민 결합 단편, 또는 면역글로불린 분자의 단편을 지칭한다. 융합 파트너는 중쇄 면역글로불린의 Fc 도메인과 같은 올리고머화 도메인을 포함할 수 있다.
"단편 결정화가능한(Fc) 폴리펩타이드"는 효과기 분자 및 세포와 상호작용하는 항체 분자의 부위이다. 이는 면역글로불린 중쇄의 C-말단 부위를 포함한다. 본원에 사용된 바와 같이, Fc 폴리펩타이드는 전체 Fc 폴리펩타이드의 하나 이상의 생물학적 활성을 지닌 Fc 도메인의 단편을 포함한다. Fc 폴리펩타이드의 "효과기 기능"은 자극에 대한 반응시 임의의 항체에 의해 전체적으로 또는 부분적으로 수행된 작용 또는 활성이며 보체 고정 또는 ADCC(항체-의존성 세포 세포독성) 유도를 포함할 수 있다.
용어 "IgX" 또는 "IgX Fc"는 특수한 항체 동형(예컨대, IgG, IgA, IgD, IgE, IgM 등)으로부터 유래된 Fc 영역을 의미하며, 여기서 "X"는 항체 동형을 나타낸다. 따라서, "IgG" 또는 "IgG Fc"는 γ 쇄의 Fc 영역을 나타내고, "IgA" 또는 "IgA Fc"는 α 쇄의 Fc 영역을 나타내고, "IgD" 또는 "IgD Fc"는 δ 쇄의 Fc 영역을 나타내고, "IgE" 또는 "IgE Fc"는 ε 쇄의 Fc 영역을 나타내며, "IgM" 또는 "IgM Fc"는 μ 쇄의 Fc 영역 등을 나타낸다. 일부 구현예에서, IgG Fc 영역은 CH1, 힌지(hinge), CH2, CH3, 및 CL1을 포함한다. "IgXN" 또는 "IgXN Fc"는 Fc 영역이 항체 동형의 특수한 소부류(예를 들면, 개과 IgG 소부류 A, B, C, 또는 D; 고양이과 IgG 소부류 1, 2a, 또는 2b; 또는 말과 IgG 소부류 IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, 또는 IgG7 등)로부터 유래됨을 나타내며, 여기서 "N"은 소부류를 나타낸다.
일부 구현예에서, IgX 또는 IgXN 영역은 반려 동물, 예를 들면, 개, 고양이, 또는 말로부터 유래된다. 일부 구현예에서, IgG 영역은 개과 γ 중쇄, 예를 들면, IgGA, IgGB, IgGC, 또는 IgGD로부터 단리된다. 일부 예에서, IgG Fc 영역은 고양이과 γ 중쇄, 예를 들면, IgG1, IgG2a, 또는 IgG2b로부터 단리된다. 다른 예에서, IgG 영역은 말과 γ 중쇄, 예를 들면, IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, 또는 IgG7로부터 단리된다. IgGA, IgGB, IgGC, 또는 IgGD의 Fc 영역을 포함하는 폴리펩타이드는 재조합 생산 시스템에서 보다 높은 발현 수준을 제공할 수 있다.
"신호 서열"은 아미노산 잔기 또는 이를 암호화하는 폴리뉴클레오타이드의 서열을 지칭하며, 이는 목적한 폴리펩타이드의 분비를 촉진하고 전형적으로 세포 표면 막의 외부로 폴리펩타이드의 배출 시 절단된다.
"링커"는 제1의 폴리펩타이드를 제2의 폴리펩타이드와 연결시키는 하나 이상의 아미노산 잔기를 지칭한다.
일부 구현예에서, 링커는 글리신이 풍부한(glycine-rich), 세린이 풍부한(serine-rich), 또는 GS가 풍부한(GS-rich) 굴곡성의 비-구조적 링커이다. 일부 구현예에서, 링커는 아미노산 G(Gly) 및/또는 S(Ser)를 포함한다. 예를 들면, 링커는 G 또는 G의 반복체(예컨대, GG, GGG 등); GS 또는 GS의 반복체(예컨대, GSGS(서열 번호: 38), GSGSGS (서열 번호: 39) 등); GGS 또는 GGS의 반복체(예컨대, GGSGGS(서열 번호: 40), GGSGGSGGS(서열 번호: 41) 등); GGGS(서열 번호: 42) 또는 GGGS의 반복체(서열 번호: 42)(예컨대, GGGSGGGS(서열 번호: 43), GGGSGGGSGGGS(서열 번호: 44) 등); GSS 또는 GSS의 반복체(예컨대, GSSGSS(서열 번호: 45), GSSGSSGSS(서열 번호: 46) 등); 또는 GGSS(서열 번호: 47) 또는 GGSS의 반복체(서열 번호: 47)(예컨대, GGSSGGSS(서열 번호: 48), GGSSGGSSGGSS(서열 번호: 49) 등)을 포함한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 적어도 하나의 융합 파트너 및/또는 적어도 하나의 링커를 포함한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 임의의 신호 서열, 임의의 융합 파트너, 및 적어도 하나의 임의의 링커를 포함한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 신호 서열, 융합 파트너, 또는 링커를 포함하지 않는다. 일부 구현예에서, IL13/IL4 연속된 폴리펩타이드는 신호 서열로 해독되지만, 신호 서열은 IL13R/IL4R 연속된 폴리펩타이드로부터 절단된다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는:
식 I: IL13R-L1-IL4R-L2-FP 또는
식 II: IL4R-L1-IL13R-L2-FP를 포함하고,
여기서 IL13R은 반려 동물 종으로부터 유래된 IL13R 세포외 도메인(ECD) 폴리펩타이드이고, IL4R은 반려 동물 종으로부터 유래된 IL4R ECD 폴리펩타이드이며, L1은 제1의 임의의 링커이고, L2는 제2의 임의의 링커이며, FP는 임의의 융합 파트너이다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 서열 번호: 13, 서열 번호: 14, 서열 번호: 15, 서열 번호: 16, 서열 번호: 17, 서열 번호: 18, 서열 번호: 19, 서열 번호: 20, 서열 번호: 21, 서열 번호: 28, 서열 번호: 29, 서열 번호: 30, 서열 번호: 31로부터 선택된 아미노산 서열을 포함한다.
IL13R/IL4R 연속된 폴리펩타이드 발현 및 생산
신호 서열이 존재하거나 부재하는 IL13R/ IL4R 연속된 폴리펩타이드의 모두 또는 일부(예컨대, 세포외 도메인)를 암호화하는 폴리뉴클레오타이드 서열이 제공된다. 상동성 신호 서열(즉, 천연의 IL-4R 또는 IL13R의 신호 서열)이 핵산 분자의 작제시 사용되지 않은 경우, 다른 신호 서열, 예를 들면, 제PCT US06/02951호에 기술된 신호 서열 중 임의의 하나를 사용할 수 있다.
전형적으로, 목적한 폴리펩타이드, 예를 들면, IL13R/IL4R 연속된 폴리펩타이드를 암호화하는 뉴클레오타이드 서열을 선택된 숙주 세포내에서 발현시키기에 적합한 발현 벡터 내로 삽입한다.
"벡터"는 하나의 유기체로부터의 DNA 서열을 다른 것에 이동시키거나 목적한 유전자를 발현시키는데 사용될 수 있는 플라스미드이다. 벡터는 전형적으로 복제 오리진(origin) 및 목적한 유전자의 발현을 조절하는 조절 서열을 포함하며, 선택적인 마커 유전자, 예를 들면, 항생제 내성 유전자를 수반하거나 수반하지 않을 수 있다. 벡터는 이것이 발현되는 숙주 세포에 적합하다. 벡터는 목적한 유전자가 벡터내에 존재하는 경우 "재조합 벡터"로 명명될 수 있다.
"숙주 세포"는 벡터 또는 단리된 폴리뉴클레오타이드의 수용체일 수 있거나 수용체인 세포를 지칭한다. 숙주세포는 원핵 세포 또는 진핵 세포일 수 있다. 예시적인 진핵 세포는 포유동물 세포, 예를 들면, 영장류 또는 비-영장류 동물 세포; 진균 세포, 예를 들면, 효모; 식물 세포; 및 곤충 세포일 수 있다. 비제한적인 예시적인 포유동물 세포는 NS0 세포, PER.C6® 세포(Crucell), 293 세포, 및 CHO 세포, 및 이들의 유도체, 예를 들면, 293-6E, DG44, CHO-S, 및 CHO-K 세포를 포함하나, 이에 한정되지 않는다. 숙주 세포는 단일의 숙주 세포의 후대세포를 포함하며, 후대세포는 천연적인, 우연한, 또는 유리된 돌연변이로 인하여 원래의 모 세포에 대해 필수적으로 완전히 동일하지 않을 수 있다(형태학적으로 또는 게놈 DNA 보체에 있어서). 숙주 세포는 본원에 제공된 아미노산 서열(들)을 암호화하는 폴리뉴클레오타이드(들)로 생체내에서 형질감염된 세포를 포함한다.
본원에 사용된 바와 같이 용어 "단리된"은 이를 사용하여 전형적으로 천연에서 발견되거나 생산되는 성분들 중 적어도 일부로부터 분리된 분자를 지칭한다. 예를 들면, 폴리펩타이드는 이것이 생산된 세포의 성분들 중 적어도 일부로부터 분리되는 경우 "단리된"으로 지칭된다. 폴리펩타이드가 발현 후 세포에 의해 분비되는 경우, 이를 생산한 세포로부터의 폴리펩타이드를 함유하는 상층액을 물리적으로 분리하는 것은 폴리펩타이드를 "단리"하는 것으로 고려된다. 유사하게, 폴리뉴클레오타이드는 이것이 전형적으로 천연에서 발견되는 보다 큰 폴리뉴클레오타이드(예를 들면, DNA 폴리뉴클레오타이드의 경우에, 게놈성 DNA 또는 미토콘드리아 DNA와 같음)의 일부가 아닌 경우, 또는 예를 들면, RNA 폴리뉴클레오타이드의 경우에, 이것이 생산된 세포의 성분들 중 적어도 일부로부터 분리되는 경우, "단리된"으로 지칭된다. 따라서, 숙주 세포 내부의 벡터내에 함유된 DNA 폴리뉴클레오타이드는 "단리된"으로서 지칭될 수 있다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 크로마토그래피, 예를 들면, 크기 배제 크로마토그래피, 이온 교환 크로마토그래피, 단백질 A 컬럼 크로마토그래피, 소수성 상호작용 크로마토그래피, 및 CHT 크로마토그래피를 사용하여 단리시킨다.
용어 "표지" 및 "검출가능한 표지"는 IL13R/IL4R 연속된 폴리펩타이드에 부착하여 이것이 검출가능하게 되도록 하는 모이어티(moiety)를 의미한다. 일부 구현예에서, 표지는 가시적인 또는 장치적 수단, 예를 들면, 방사표지된 아미노산의 혼입 또는 표지된 아비딘(예를 들면, 광학적 또는 열량계적 방법에 의해 측정될 수 있는 형광성 마커 또는 효소적 활성을 함유하는 스트렙타비딘)에 의해 검출될 수 있는 바이오티닐 모이어티의 폴리펩타이드에 대한 부착에 의해 검출될 수 있는 신호를 생성하는 검출 가능한 마커이다. 폴리펩타이드에 대한 표지의 예는 다음을 포함하나, 이에 한정되지 않는다: 방사성동위원소 또는 방사성 핵종(예를 들면, 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, 또는 153Sm); 크로모겐, 형광성 표지(예를 들면, FITC, 로다민, 란타나이드 포스포르), 효소 표지(예를 들면, 서양고추냉이 퍼옥시다제, 루시퍼라제, 알칼린 포스파타제); 화학발광성 마커; 바이오티닐 그룹; 제2 리포터(예를 들면, 루이신 지퍼 쌍 서열(leucine zipper pair sequence), 제2의 항체에 대한 결합 부위, 금속 결합 도메인, 에피토프 태그(epitope tag))에 의해 인식된 예정된 폴리펩타이드 에피토프; 및 자기 제제(magnetic agent), 예를 들면, 고돌리늄 킬레이트. 면역검정에 일반적으로 사용된 표지의 대표적인 예는 광을 생산하는 모이어티, 예를 들면, 아크리디늄 화합물, 및 형광성을 생산하는 모이어티, 예를 들면, 플루오레세인을 포함한다. 이와 관련하여, 모이어티 자체는 검출가능하게 표지되지 않을 수 있지만 여전히 다른 모이어티와 반응시 검출가능하게 될 수 있다.
데코이 수용체 트랩(Decoy Receptor Trap)으로서 IL13R/IL4R 연속된 폴리펩타이드
본 발명의 IL13R/IL4R 연속된 폴리펩타이드는 IL13 또는 IL4를 트랩핑(trapping)하고 세포 표면에서 IL13R 및 IL4R과 이의 상호작용을 억제하는 데코이 수용체로서 기능할 수 있다. 데코이 수용체, 예를 들어, 본 발명의 것은 이들의 리간드를 고 친화성 및 특이성으로 인식하지만 구조적으로 신호전달할 수 없다. 이들은 리간드 결합에 대해 야생형 수용체와 경쟁하며 리간드/수용체 상호작용에 관여하므로, 기능화 수용체의 활성 또는 수 및/또는 수용체로부터 하부로 세포 활성을 조절한다. 데코이 수용체는 효능제 리간드에 대한 분자 트랩으로서 작용할 수 있으며 이에 의해 리간드-유도된 수용체 활성화를 억제한다.
본원에 사용된 바와 같이 "IL13"은 세포내에서 IL13의 발현 및 가공으로 생성되는 임의의 천연 IL13을 지칭한다. 이러한 용어는 달리 나타내지 않는 한, 임의의 척추동물원, 예를 들면, 포유동물, 예를 들면, 영장류(예컨대, 사람 및 시노몰구스 원숭이) 및 설치류(예컨대, 마우스 및 랫트), 및 반려 동물(예컨대, 개, 고양이, 및 말)로부터의 IL13을 포함한다. 이러한 용어는 또한 IL13의 천연적으로 발생하는 변이체, 예컨대, 스플라이스 변이체(splice variant) 또는 대립형질 변이체(allelic variant)를 포함한다.
일부 구현예에서, 개과 IL13은 서열 번호: 4의 아미노산 서열을 포함한다. 일부 구현예에서, 고양이과 IL13은 서열 번호: 5의 아미노산 서열을 포함한다. 일부 구현예에서, 말과 IL13은 서열 번호: 6의 아미노산 서열을 포함한다.
본원에 사용된 바와 같이 "IL4"는 세포내에서 IL4의 발현 및 가공을 야기하는 임의의 천연 IL4를 지칭한다. 이러한 용어는 달리 나타내지 않는 한, 임의의 척추동물원, 예를 들면, 포유동물, 예를 들면, 영장류(예컨대, 사람 및 시노몰구스 원숭이) 및 설치류(예컨대, 마우스 및 랫트), 및 반려 동물(예컨대, 개, 고양이, 및 말)로부터의 IL4를 포함한다. 이러한 용어는 또한 IL4의 천연적으로 발생하는 변이체, 예컨대, 스플라이스 변이체 또는 대립형질 변이체를 포함한다.
일부 구현예에서, 개과 IL4는 서열 번호: 1의 아미노산 서열을 포함한다. 일부 구현예에서, 고양이과 IL4는 서열 번호: 2의 아미노산 서열을 포함한다. 일부 구현예에서, 말과 IL4는 서열 번호: 3의 아미노산 서열을 포함한다.
본 발명은 치료제로서 IL13R/IL4R 연속된 폴리펩타이드를 제공한다. 본 발명의 IL13R/IL4R 연속된 폴리펩타이드는 본원에 보다 상세히 기술된, IL13 및/또는 IL4에 결합하며, 이는 알레르기 질환과 관련된 것으로 입증되었다. 다양한 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 IL13 및 IL4과 매우 높은 친화성으로 결합할 수 있다. 다양한 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 IL13 및 IL4 신호전달을 방해할 수 있다.
용어 "친화성"은 분자(예컨대, 수용체)의 단일 결합 부위와 이의 결합 파트너(예를 들면, 리간드) 사이의 비공유결합성 상호작용의 총 합의 강도를 의미한다. 이의 파트너 Y에 대한 분자 X의 친화성은 일반적으로 해리 상수(KD)로 나타낸다. 친화성은 당해 분야에 공지된 일반적인 방법, 예를 들면, 면역블롯(immunoblot), ELISA KD, KinEx A, 생물층 간섭법(biolayer interferometry: BLI), 또는 표면 플라스몬 공명 장치에 의해 측정할 수 있다.
용어 "KD," "Kd," "Kd" 또는 "Kd 값"은 수용체 융합-리간드 상호작용의 평형 해리 상수를 지칭하기 위해 상호교환적으로 사용된다. 일부 구현예에서, 이의 리간드에 대한 융합 분자의 Kd는 바이오센서, 예를 들면, Octet® 시스템(Pall ForteBio LLC, 캘리포니아주 프레몬트(Fremont) 소재)을 공급업자의 지시사항에 따라 사용하는 생물층 간섭법 검정을 사용함으로써 측정된다. 요약하면, 바이오티닐화된 항원을 센서 팁에 결합시키고 융합 분자의 해리를 90초 동안 모니터링하고 해리를 600초 동안 모니터링한다. 희석 및 결합 단계용 완충제는 20 mM 포스페이트, 150 mM NaCl, pH 7.2이다. 완충제 만의 블랭크 곡선(blank curve)은 임의의 이동(drift)을 교정하기 위해 감해진다. 데이타는 ForteBio 분석 소프트웨어를 사용하여 2:1 결합 모델에 대해 조절함으로써 결합 속도 상수(kon), 해리 속도 상수(koff), 및 Kd를 측정한다. 평형 해리 상수(Kd)는 koff/kon의 비로서 계산된다. 용어 "kon"은 이의 파트너 Y에 대한 분자 X의 결합에 대한 속도 상수를 지칭하고 용어 "koff"는 분자 X/파트너 Y 복합체로부터 분자 X 또는 파트너 Y의 해리에 대한 속도 상수를 지칭한다.
용어 물질에 "결합하다"는 당해 분야에 잘 이해되고 있는 용어이며, 이러한 결합을 측정하는 방법은 또한 당해 분야에 잘 공지되어 있다. 분자는 특수한 세포 또는 물질에 대해 반응하거나, 이와 결합되거나, 이에 대해 친화성을 갖는 경우 "결합"을 나타내는 것으로 일컬어지며, 반응, 결합, 또는 친화성은 당해 분야에 공지된 하나 이상의 방법, 예를 들면, 면역블롯, ELISA KD, KinEx A, 생물층 간섭계(BLI), 표면 플라스몬 공명 장치 등에 의해 검출될 수 있다.
"표면 플라스몬 공명"은 바이오센서 매트릭스, 예를 들면 BIAcoreTM 시스템(BIAcore International AB, GE Healthcare company, 스웨덴 업살라(Uppsala) 및 뉴저지주 피스카타웨이 소재) 내에서 단백질 농도에 있어서의 변경을 검출함으로써 실시간 생물특이적인 상호작용의 분석을 가능하도록 하는 광학적 현상을 나타낸다. 추가의 설명에 대해서는, 문헌: Jonsson et al. (1993) Ann. Biol. Clin. 51: 19-26을 참고한다.
"생물층 간섭법"은 바이오센서 팁 및 내부 참고 층 상에서 고정된 단백질의 층으로부터 반사된 광의 간섭 패턴을 분석하는 광학 분석 기술을 지칭한다. 바이오센서 팁에 결합된 분자의 수에 있어서의 변화는 실시간으로 측정될 수 있는 간섭 패턴에 있어서의 이동을 유발한다. 바이오층 간섭법용의 비제한적인 예시적인 장치는 문헌: Octet® 시스템(Pall ForteBio LLC)이다. 예컨대, Abdiche et al., 2008, Anal. Biochem. 377: 209-277을 참고한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 생물층 간섭법으로 측정된 바와 같이, 개과 IL13 및/또는 IL4, 고양이과 IL13 및/또는 IL4, 또는 말과 IL13 및/또는 IL4에 5 x 10-6 M 미만, 1 x 10-6 M 미만, 5 x 10-7 M 미만, 1 x 10-7 M 미만, 5 x 10-8 M 미만, 1 x 10-8 M 미만, 5 x 10-9 M 미만, 1 x 10-9 M 미만, 5 x 10-10 M 미만, 1 x 10-10 M 미만, 5 x 10-11 M 미만, 1 x 10-11 M 미만, 5 x 10-12 M 미만, 또는 1 x 10-12 M 미만의 해리 상수(Kd)로 결합한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 생물층 간섭법으로 측정된 바와 같이, 개과 IL13 및/또는 IL4, 고양이과 IL13 및/또는 IL4, 또는 말과 IL13 및/또는 IL4에 5 x 10-6 M 내지 1 x 10-6 M, 5 x 10-6 M 내지 5 x 10-7 M, 5 x 10-6 M 내지 1 x 10-7 M, 5 x 10-6 M 내지 5 x 10-8 M, 5 x 10-6 M 내지 1 x 10-8 M, 5 x 10-6 M 내지 5 x 10-9 M, 5 x 10-6 M 내지 1 x 10-9 M, 5 x 10-6 M 내지 5 x 10-10 M, 5 x 10-6 M 내지 1 x 10-10 M, 5 x 10-6 M 내지 5 x 10-11 M, 5 x 10-6 M 내지 1 x 10-11 M, 5 x 10-6 M 내지 5 x 10-12 M, 5 x 10-6 M 내지 1 x 10-12 M, 1 x 10-6 M 내지 5 x 10-7 M, 1 x 10-6 M 내지 1 x 10-7 M, 1 x 10-6 M 내지 5 x 10-8 M, 1 x 10-6 M 내지 1 x 10-8 M, 1 x 10-6 M 내지 5 x 10-9 M, 1 x 10-6 M 내지 1 x 10-9 M, 1 x 10-6 M 내지 5 x 10-10 M, 1 x 10-6 M 내지 1 x 10-10 M, 1 x 10-6 M 내지 5 x 10-11 M, 1 x 10-6 M 내지 1 x 10-11 M, 1 x 10-6 M 내지 5 x 10-12 M, 1 x 10-6 M 내지 1 x 10-12 M, 5 x 10-7 M 내지 1 x 10-7 M, 5 x 10-7 M 내지 5 x 10-8 M, 5 x 10-7 M 내지 1 x 10-8 M, 5 x 10-7 M 내지 5 x 10-9 M, 5 x 10-7 M 내지 1 x 10-9 M, 5 x 10-7 M 내지 5 x 10-10 M, 5 x 10-7 M 내지 1 x 10-10 M, 5 x 10-7 M 내지 5 x 10-11 M, 5 x 10-7 M 내지 1 x 10-11 M, 5 x 10-7 M 내지 5 x 10-12 M, 5 x 10-7 M 내지 1 x 10-12 M, 1 x 10-7 M 내지 5 x 10-8 M, 1 x 10-7 M 내지 1 x 10-8 M, 1 x 10-7 M 내지 5 x 10-9 M, 1 x 10-7 M 내지 1 x 10-9 M, 1 x 10-7 M 내지 5 x 10-10 M, 1 x 10-7 M 내지 1 x 10-10 M, 1 x 10-7 M 내지 5 x 10-11 M, 1 x 10-7 M 내지 1 x 10-11 M, 1 x 10-7 M 내지 5 x 10-12 M, 1 x 10-7 M 내지 1 x 10-12 M, 5 x 10-8 M 내지 1 x 10-8 M, 5 x 10-8 M 내지 5 x 10-9 M, 5 x 10-8 M 내지 1 x 10-9 M, 5 x 10-8 M 내지 5 x 10-10 M, 5 x 10-8 M 내지 1 x 10-10 M, 5 x 10-8 M 내지 5 x 10-11 M, 5 x 10-8 M 내지 1 x 10-11 M, 5 x 10-8 M 내지 5 x 10-12 M, 5 x 10-8 M 내지 1 x 10-12 M, 1 x 10-8 M 내지 5 x 10-9 M, 1 x 10-8 M 내지 1 x 10-9 M, 1 x 10-8 M 내지 5 x 10-10 M, 1 x 10-8 M 내지 1 x 10-10 M, 1 x 10-8 M 내지 5 x 10-11 M, 1 x 10-8 M 내지 1 x 10-11 M, 1 x 10-8 M 내지 5 x 10-12 M, 1 x 10-8 M 내지 1 x 10-12 M, 5 x 10-9 M 내지 1 x 10-9 M, 5 x 10-9 M 내지 5 x 10-10 M, 5 x 10-9 M 내지 1 x 10-10 M, 5 x 10-9 M 내지 5 x 10-11 M, 5 x 10-9 M 내지 1 x 10-11 M, 5 x 10-9 M 내지 5 x 10-12 M, 5 x 10-9 M 내지 1 x 10-12 M, 1 x 10-9 M 내지 5 x 10-10 M, 1 x 10-9 M 내지 1 x 10-10 M, 1 x 10-9 M 내지 5 x 10-11 M, 1 x 10-9 M 내지 1 x 10-11 M, 1 x 10-9 M 내지 5 x 10-12 M, 1 x 10-9 M 내지 1 x 10-12 M, 5 x 10-10 M 내지 1 x 10-10 M, 5 x 10-10 M 내지 5 x 10-11 M, 1 x 10-10 M 내지 5 x 10-11 M, 1 x 10-10 M 내지 1 x 10-11 M, 1 x 10-10 M 내지 5 x 10-12 M, 1 x 10-10 M 내지 1 x 10-12 M, 5 x 10-11 M 내지 1 x 10-12 M, 5 x 10-11 M 내지 5 x 10-12 M, 5 x 10-11 M 내지 1 x 10-12 M, 1 x 10-11 M 내지 5 x 10-12 M, 또는 1 x 10-11 M 내지 1 x 10-12 M의 Kd로 결합한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 개과 IL13 및/또는 IL4, 고양이과 IL13 및/또는 IL4, 및/또는 말과 IL13 및/또는 IL4에 결합한다.
"감소시키다" 또는 "억제하다"는 참고와 비교하여 활성, 기능, 또는 양을 저하시키거나, 감소시키거나 저지함을 의미한다. 일부 구현예에서, "감소시키다" 또는 "억제하다"는 20% 이상의 전체적인 감소를 유발하는 능력을 의미한다. 일부 구현예에서, "감소시키다" 또는 "억제하다"는 50% 이상의 전체적인 감소를 유발하는 능력을 의미한다. 일부 구현예에서, "감소시키다" 또는 "억제하다"는 75%, 85%, 90%, 95% 이상의 전체적인 감소를 유발하는 능력을 의미한다. 일부 구현예에서, 상기 나타낸 양은 동일한 기간에 걸쳐서 대조군 용량(예를 들면, 위약)에 대하여, 기간에 걸쳐 억제되거나 감소된다. 본원에 사용된 바와 같이, "참고"는 비교 목적을 위해 사용된 임의의 샘플, 표준, 또는 수준을 지칭한다. 참고는 건강하거나 질환이 없는 샘플로부터 수득될 수 있다. 일부 예에서, 참고는 반려 동물의 질환이 없거나 치료되지 않은 반려 동물 샘플로부터 수득된다. 일부 예에서, 참고는 특수한 종의 하나 이상의 건강한 동물로부터 수득되며, 이는 치료되고 있거나 치료된 동물이 아니다.
본원에 사용된 바와 같이, 용어 "실질적으로 감소된"은 수치와 참고 수치 사이의 충분히 높은 정도의 감소를 나타냄으로써 당해 분야의 기술자는 2개 값들 사이의 차이가 상기 값에 의해 측정된 생물학적 특징의 맥락내에 통계적인 유의성이 있는 것으로 고려될 수 있다. 일부 구현예에서, 실질적으로 감소된 수치는 참고 값과 비교하여 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 또는 100% 중 임의의 하나보다 더 높게 감소된다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 융합 분자의 부재하에서 IL13 및/또는 IL4 신호전달과 비교하여, 반려 동물 종 내 IL13 및/또는 IL4 신호전달을 적어도 10%, 적어도 15%, 적어도 20%, 적어도 25%, 적어도 30%, 적어도 35%, 적어도 40%, 적어도 45%, 적어도 50%, 적어도 60%, 적어도 70%, 적어도 80%, 적어도 90%, 또는 100%까지 감소시킬 수 있다. 일부 구현예에서, 신호전달은 IL4-의존성 TF-1 세포 증식에 있어서의 감소에 의해 측정된다. 일부 구현예에서, IL13 및/또는 IL4 신호전달에 있어서의 감소 또는 증식에 있어서의 감소는 10% 내지 15%, 10% 내지 20%, 10% 내지 25%, 10% 내지 30%, 10% 내지 35%, 10% 내지 40%, 10% 내지 45%, 10% 내지 50%, 10% 내지 60%, 10% 내지 70%, 10% 내지 80%, 10% 내지 90%, 10% 내지 100%, 15% 내지 20%, 15% 내지 25%, 15% 내지 30%, 15% 내지 35%, 15% 내지 40%, 15% 내지 45%, 15% 내지 50%, 15% 내지 60%, 15% 내지 70%, 15% 내지 80%, 15% 내지 90%, 15% 내지 100%, 20% 내지 25%, 20% 내지 30%, 20% 내지 35%, 20% 내지 40%, 20% 내지 45%, 20% 내지 50%, 20% 내지 60%, 20% 내지 70%, 20% 내지 80%, 20% 내지 90%, 20% 내지 100%, 25% 내지 30%, 25% 내지 35%, 25% 내지 40%, 25% 내지 45%, 25% 내지 50%, 25% 내지 60%, 25% 내지 70%, 25% 내지 80%, 25% 내지 90%, 25% 내지 100%, 30% 내지 35%, 30% 내지 40%, 30% 내지 45%, 30% 내지 50%, 30% 내지 60%, 30% 내지 70%, 30% 내지 80%, 30% 내지 90%, 30% 내지 100%, 35% 내지 40%, 35% 내지 45%, 35% 내지 50%, 35% 내지 60%, 35% 내지 70%, 35% 내지 80%, 35% 내지 90%, 35% 내지 100%, 40% 내지 45%, 40% 내지 50%, 40% 내지 60%, 40% 내지 70%, 40% 내지 80%, 40% 내지 90%, 40% 내지 100%, 45% 내지 50%, 45% 내지 60%, 45% 내지 70%, 45% 내지 80%, 45% 내지 90%, 45% 내지 100%, 50% 내지 60%, 50% 내지 70%, 50% 내지 80%, 50% 내지 90%, 50% 내지 100%, 60% 내지 70%, 60% 내지 80%, 60% 내지 90%, 60% 내지 100%, 70% 내지 80%, 70% 내지 90%, 70% 내지 100%, 80% 내지 90%, 80% 내지 100%, 또는 90% 내지 100%이다.
약제학적 조성물
용어 "약제학적 제형" 및 "약제학적 조성물"은 이러한 형태에서 활성 성분(들)의 생물학적 활성이 효과적이도록 하고, 제형이 투여될 수 있는 대상체에 대해 허용불가능하게 독성인 추가의 성분을 함유하지 않는 제제를 지칭한다.
"약제학적으로 허용되는 담체"는 대상체에게 투여하기 위한 "약제학적 조성물"을 함께 포함하는 치료제와 함께 사용하기 위한 당해 분야에서 통상적인 비-독성 고체, 반고체 또는 액체 충전제, 희석제, 캘슐화 물질, 제형 보조제, 또는 담체를 지칭한다. 약제학적으로 허용되는 담체는 사용된 투여량 및 농도에서 수용체에 대해 비-독성이고 제형의 다른 성분과 혼화성이다. 약제학적으로 허용되는 담체는 사용된 제형에 대해 적절하다. 약제학적으로 허용되는 담체의 예는 알루미나; 스테아르산알루미늄; 레시틴; 혈청 단백질, 예를 들면, 사람 혈청 알부민, 개과 또는 다른 동물 알부민; 완충제, 예를 들면, 포스페이트, 시트레이트, 트로메타민 또는 HEPES 완충제; 글리신; 소르브산; 소르브산칼륨; 포화된 식물성 지방산의 부분 글리세라이드 혼합물; 물; 염 또는 전해질, 예를 들면, 프로타민 설페이트, 인산수소이나트륨, 인산수소칼륨, 염화나트륨, 아연 염, 콜로이드성 실리카, 또는 마그네슘 트리실리케이트; 폴리비닐 피롤리돈, 셀룰로즈-계 물질; 폴리에틸렌 글리콜; 슈크로즈; 만니톨; 또는 아르기닌을 포함하나, 이에 한정되지 않는 아미노산을 포함한다.
약제학적 조성물은 동결건조된 형태로 저장될 수 있다. 따라서, 일부 구현예에서, 제조 공정은 동결건조 단계를 포함한다. 동결건조된 조성물을 이후에 전형적으로 개, 고양이, 또는 말에게 투여하기 전에 비경구 투여용으로 적합한 수성 조성물로서 재제형화할 수 있다. 다른 구현예에서, 특히 융합 분자가 열 및 산화성 변성에 매우 안정한 경우, 약제학적 조성물은 액체로서, 즉, 수성 조성물로서 저장될 수 있으며, 이는 직접, 또는 적절한 희석하에 개, 고양이, 또는 말에게 투여될 수 있다. 동결건조된 조성물은 멸균 주사용수(WFI)로 재구성할 수 있다. 제균 시약(bacteriostatic reagent), 예를 들면, 벤질 알코올이 포함될 수 있다. 따라서, 본 발명은 고체 또는 액체형의 약제학적 조성물을 제공한다.
약제학적 조성물의 pH는 투여하는 경우 약 pH 5 내지 약 pH 8의 범위내일 수 있다. 본 발명의 조성물은 치료학적 목적으로 사용될 경우 멸균성이다. 멸균성은 당해 분야에 공지된 임의의 수개의 수단, 예를 들면, 멸균 여과 막(예컨대, 0.2 마이크론 막)을 통한 여과에 의해 달성될 수 있다. 멸균성은 항세균제의 존재하에 또는 부재하에서 유지될 수 있다.
IL13R/IL4R 연속된 폴리펩타이드 및 약제학적 조성물의 용도
IL13R/IL4R 연속된 폴리펩타이드 또는 본 발명의 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물은 IL13- 및/또는 IL4-유도된 상태를 치료하는데 유용할 수 있다. 본원에 사용된 바와 같이, "IL13 또는 IL4-유도된 상태"는 IL13 또는 IL4의 상승된 수준 또는 변경된 분포와 관련되거나, 이에 의해 유발되거나, 이에 의해 특징화된 질환을 의미한다. 이러한 IL13 및/또는 IL4-유도된 상태는 소양성 또는 알레르기 질환을 포함하나, 이에 한정되지 않는다. 일부 구현예에서, IL13- 및/또는 IL4-유도된 상태는 아토피성 피부염, 소양증, 천식, 건선, 피부경화증, 또는 습진이다. IL13- 또는 IL4-유도된 상태는 반려동물, 예를 들면, 개과, 고양이과, 또는 말과를 포함하나 이에 한정되지 않는 반려동물에서 나타날 수 있다.
본원에 사용된 바와 같이, "치료"는 유리하거나 목적한 임상 결과를 수득하기 위한 접근법이다. 본원에 사용된 바와 같이, "치료"는 반려 동물을 포함하는 포유동물에서 질환에 대한 치료제의 임의의 투여 또는 적용을 포함한다. 본 개시내용의 목적을 위해, 유리한 또는 목적한 임상 결과는 다음 중 임의의 하나 이상을 포함하나, 이에 한정되지 않는다: 하나 이상의 증상의 완화, 질환 정도의 약화, 질환의 확산 방지 또는 지연, 질환의 재발 방지 또는 지연, 질환 진행의 지연 또는 늦춤, 질환 상태의 경감, 질환 또는 질환 진행의 억제, 질환 또는 이의 진행의 억제 또는 늦춤, 이의 발달의 정지, 및 차도(부분적으로 또는 천체적으로). 또한 "치료"는 증식성 질환의 병리학적 결과의 감소를 포함한다. 본원에 제공된 방법은 치료의 이러한 양태 중 임의의 하나 이상을 고려한다. 상기와 일치하여, 용어 치료는 장애의 모든 양태의 100% 제거를 필요로 하지 않는다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 이를 포함하는 약제학적 조성물은 IL13- 또는 IL4-유도된 상태를 치료하기 위해 본원의 방법에 따라 이용될 수 있다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물은 반려 동물, 예를 들면, 개과, 고양이과, 또는 말과에 투여되어, IL13- 및 IL4-유도된 상태를 치료한다.
물질/분자, 효능제 또는 길항제의 "치료학적 유효량"은 치료될 질환의 유형, 질환 상태, 질환의 중증도 및 과정, 치료 목적의 유형, 임의의 이전 치료요법, 임상 병력, 사전 치료에 대한 반응, 주치 수의사의 판단, 동물의 연령, 성별, 및 체중, 및 동물에서 목적한 반응을 유발하는 물질/분자, 효능제 또는 길항제의 능력과 같은 인자에 따라 변할 수 있다. 치료학적 유효량은 또한 물질/분자, 효능제 또는 길항제의 임의의 독성 또는 유해한 효과보다 치료학적으로 유리한 효과가 더 큰 것이다. 치료학적 유효량은 하나 이상의 투여로 전달될 수 있다. 치료학적 유효량은 목적한 치료학적 또는 예방학적 결과를 달성하는데 필요한 용량에서 및 기간 동안 효과적인 양을 지칭한다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물은 피하 투여, 정맥내 주입, 또는 근육내 주사에 의해 비경구적으로 투여된다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물은 거환 주사(bolus injection)로서 또는 연속 주입에 의해 일정 기간에 걸쳐 투여된다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물은 근육내, 복강내, 뇌척수내, 피하, 동맥내, 활액내, 척추강내, 또는 흡입 경로에 의해 투여된다.
본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드는 용량당 0.1 mg/kg의 체중 내지 100 mg/kg의 체중의 범위의 양으로 투여될 수 있다. 일부 구현예에서, 항- IL13/IL4 융합체는 용량당 0.5 mg/kg의 체중 내지 50 mg/kg의 체중의 범위의 양으로 투여될 수 있다. 일부 구현예에서, 항-IL13/IL4 융합체는 용량당 1 mg/kg의 체중 내지 10 mg/kg의 체중의 범위의 양으로 투여될 수 있다. 일부 구현예에서, 융합 분자는 0.5 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 1 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 5 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 10 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 20 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 50 mg/kg의 체중 내지 100 mg/kg의 체중의 범위, 1 mg/kg의 체중 내지 10 mg/kg의 체중의 범위, 5 mg/kg의 체중 내지 10 mg/kg의 체중의 범위, 0.5 mg/kg의 체중 내지 10 mg/kg의 체중, 또는 5 mg/kg의 체중 내지 50 mg/kg의 체중의 범위로 투여될 수 있다.
IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물은 반려 동물에게 1회 또는 일련의 치료에 걸쳐 투여될 수 있다. 예를 들면, IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R 및 IL4R을 포함하는 약제학적 조성물은 적어도 1회, 1회 이상, 적어도 2회, 적어도 3회, 적어도 4회, 또는 적어도 5회 투여될 수 있다.
일부 구현예에서, 용량은 적어도 2 또는 3주의 연속 중 동안 주당 1회 투여되며, 일부 구현예에서, 이러한 치료 주기는 2회 이상 반복되고, 임의로 1주 이상의 비 치료가 배치된다. 다른 구현예에서, 치료학적 유효량은 2 내지 5 연속일 동안 1일에 1회 투여되고, 일부 구현예에서, 이러한 치료 주기는 2회 이상 반복되며, 임의로 하나 이상의 수일 또는 수주의 비 치료가 배치된다.
하나 이상의 추가의 치료제 "와 함께" 투여는 동시(공동) 및 임의의 순서로 연속적이거나 순차적인 투여를 포함한다. 용어 "동시에"는 본원에서 2개 이상의 치료제의 투여를 지칭하기 위해 사용되며, 여기서 투여의 적어도 일부분은 시간내에서 중첩되거나 여기서 하나의 치료제의 투여는 다른 치료제와 비교하여 짧은 기간 내에 속한다. 예를 들면, 2개 이상의 치료제는 규정된 수의 분 이하의 시간 분리로 투여된다. 용어 "실질적으로"는 본원에서 2개 이상의 치료제의 투여를 지칭하기 위해 사용되며 여기서 하나 이상의 제제(들)의 투여는 하나 이상의 다른 제제(들)의 투여의 중지 후 지속하거나, 여기서 하나 이상의 제제(들)의 투여는 하나 이상의 다른 제제(들)의 투여 전에 시작한다. 예를 들면, 2개 이상의 치료제의 투여는 대략 규정된 수의 분보다 더 떨어진 시간으로 투여된다. 본원에 사용된 바와 같이, "~와 함께"는 다른 치료 양상외에 하나의 치료 양상의 투여를 지칭한다. 이와 같이, "~와 함께"는 동물에게의 다른 치료 양상의 투여 전, 동안 또는 후의 하나의 치료 양상의 투여를 지칭한다.
일부 구현예에서, 이러한 방법은 IL13R/IL4R 연속된 폴리펩타이드 또는, IL13R/IL4R 연속된 폴리펩타이드, Jak 억제제, PI3K 억제제, AKT 억제제, 또는 MAPK 억제제를 포함하는 약제학적 조성물과 함께 투여함을 포함한다. 일부 구현예에서, 이러한 방법은 IL13R/IL4R 연속된 폴리펩타이드 또는, IL13R/IL4R 연속된 폴리펩타이드, 항-IL17 항체, 항-TNFα 항체, 항-CD20 항체, 항-CD19 항체, 항-CD25 항체, 항-IL31 항체, 항-IL23 항체, 항-IgE 항체, 항-CD11α 항체, 항-IL6R 항체, 항-α4-인테그린 항체, 항-IL12 항체, 항-IL1β 항체, 또는 항-BlyS 항체를 포함하는 약제학적 조성물과 함께 투여함을 포함한다.
세포에 IL13R/IL4R 연속된 폴리펩타이드 또는 IL13R/IL4R 연속된 폴리펩타이드를 포함하는 약제학적 조성물을 IL13 또는 IL4에 대한 결합을 허용하는 조건 하에서 노출시키는 방법이 본원에 제공된다. 일부 구현예에서, 세포는 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 생체외에서 노출된다. 일부 구현예에서, 세포는 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 생체내에서 노출된다. 일부 구현예에서, 세포는 IL13R/IL4R 연속된 폴리펩타이드에 노출된다. 일부 구현예에서, 세포는 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 융합 단백질의 세포외 IL13 또는 IL4에 대한 결합을 허용하는 조건 하에서 노출된다. 일부 구현예에서, 세포는 본원에 기술된 임의의 하나 이상의 투여 방법, 예를 들면, 그러나 이에 한정되지 않는, 대상체내로 복강내, 근육내, 정맥내 주사에 의해 생체내에서 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 노출될 수 있다. 일부 구현예에서, 세포는 세포를 융합 분자 또는 약제학적 조성물을 포함하는 배양 배지에 노출시킴에 의해 생체외에서 IL13R/IL4R 연속된 폴리펩타이드 또는 약제학적 조성물에 노출될 수 있다. 일부 구현예에서, 세포 막의 투과성은 융합 분자 또는 약제학적 조성물을 포함하는 배양 배지에 세포를 노출시키기 전에 당해 분야의 기술자에게 의해 이해되는 임의의 수의 방법(예를 들면, 세포를 전기천공하거나 세포를 염화칼슘을 함유하는 용액에 노출시킴)을 사용하여 영향을 줄 수 있다.
일부 구현예에서, 노출 결과는 세포에 의한 IL13 또는 IL4 신호전달 기능의 감소를 야기한다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 세포내에서 IL13 또는 IL4 신호전달을 IL13R/IL4R 연속된 폴리펩타이드의 부재하에서의 IL13 또는 IL4 신호전달 기능과 비교하여 적어도 10%, 적어도 15%, 적어도 20%, 적어도 25%, 적어도 30%, 적어도 35%, 적어도 40%, 적어도 45%, 적어도 50%, 적어도 60%, 적어도 70%, 적어도 80%, 적어도 90%, 또는 100%까지 감소시킬 수 있다. 일부 구현예에서, IL13 또는 IL4 신호전달에 있어서의 감소 또는 TF-1 증식에 있어서의 감소는 10% 내지 15%, 10% 내지 20%, 10% 내지 25%, 10% 내지 30%, 10% 내지 35%, 10% 내지 40%, 10% 내지 45%, 10% 내지 50%, 10% 내지 60%, 10% 내지 70%, 10% 내지 80%, 10% 내지 90%, 10% 내지 100%, 15% 내지 20%, 15% 내지 25%, 15% 내지 30%, 15% 내지 35%, 15% 내지 40%, 15% 내지 45%, 15% 내지 50%, 15% 내지 60%, 15% 내지 70%, 15% 내지 80%, 15% 내지 90%, 15% 내지 100%, 20% 내지 25%, 20% 내지 30%, 20% 내지 35%, 20% 내지 40%, 20% 내지 45%, 20% 내지 50%, 20% 내지 60%, 20% 내지 70%, 20% 내지 80%, 20% 내지 90%, 20% 내지 100%, 25% 내지 30%, 25% 내지 35%, 25% 내지 40%, 25% 내지 45%, 25% 내지 50%, 25% 내지 60%, 25% 내지 70%, 25% 내지 80%, 25% 내지 90%, 25% 내지 100%, 30% 내지 35%, 30% 내지 40%, 30% 내지 45%, 30% 내지 50%, 30% 내지 60%, 30% 내지 70%, 30% 내지 80%, 30% 내지 90%, 30% 내지 100%, 35% 내지 40%, 35% 내지 45%, 35% 내지 50%, 35% 내지 60%, 35% 내지 70%, 35% 내지 80%, 35% 내지 90%, 35% 내지 100%, 40% 내지 45%, 40% 내지 50%, 40% 내지 60%, 40% 내지 70%, 40% 내지 80%, 40% 내지 90%, 40% 내지 100%, 45% 내지 50%, 45% 내지 60%, 45% 내지 70%, 45% 내지 80%, 45% 내지 90%, 45% 내지 100%, 50% 내지 60%, 50% 내지 70%, 50% 내지 80%, 50% 내지 90%, 50% 내지 100%, 60% 내지 70%, 60% 내지 80%, 60% 내지 90%, 60% 내지 100%, 70% 내지 80%, 70% 내지 90%, 70% 내지 100%, 80% 내지 90%, 80% 내지 100%, 또는 90% 내지 100%이다.
IL13- 또는 IL4-유도된 상태의 검출, 진단 및 모니터링을 위한 IL13R/IL4R 연속된 폴리펩타이드, 폴리펩타이드들 및 폴리뉴클레오타이드를 사용하는 방법이 본원에 제공된다. 반려 동물이 IL13R/IL4R 연속된 폴리펩타이드 치료요법에 대해 반응하는지의 여부를 측정하는 방법이 본원에 제공된다. 일부 구현예에서, 이러한 방법은 동물이 IL13R/IL4R 연속된 폴리펩타이드를 사용하여 IL13 또는 IL4를 발현하는 세포를 가지고 있는지의 여부를 검출함을 포함한다. 일부 구현예에서, 검출 방법은 샘플을 항체, 폴리펩타이드, 또는 폴리뉴클레오타이드와 접촉시키는 단계 및 결합 수준이 참고 또는 비교 샘플(예를 들면, 대조군)의 것과 상이한지의 여부를 측정하는 단계를 포함한다. 일부 구현예에서, 이러한 방법은 본원에 기술된 IL13R/IL4R 연속된 폴리펩타이드가 대상체 동물에 대해 적절한 치료인지를 측정하기 위해 사용될 수 있다.
일부 구현예에서, 샘플은 생물학적 샘플이다. 용어 "생물학적 샘플"은 살아있는 것 또는 이전에 살아있던 것으로부터의 물질의 양을 의미한다. 일부 구현예에서, 생물학적 샘플은 세포 또는 세포/조직 분해물이다. 일부 구현예에서, 생물학적 샘플은 혈액(예를 들면, 전혈), 혈장, 혈청, 뇨, 활막액, 및 상피 세포를 포함하나, 이에 한정되지 않는다.
일부 구현예에서, 세포 또는 세포/조직 분해물은 IL13R/IL4R 연속된 폴리펩타이드와 접촉되며 IL13R/IL4R 연속된 폴리펩타이드와 세포 사이의 결합이 측정된다. 시험 세포가 동일한 조직형의 참고 세포와 비교하여 결합 활성을 나타내는 경우, 이는 대상체가 IL13R/IL4R 연속된 폴리펩타이드를 사용한 치료로부터 유리할 수 있음을 나타낼 수 있다. 일부 구현예에서, 시험 세포는 반려 동물의 조직으로부터 유래한다.
특이적인 항체-항원 결합을 검출하기 위한 당해 분야에 공지된 다양한 방법을 사용할 수 있다. 수행될 수 있는 예시적인 면역검정은 형광성 편광 면역검정(FPIA), 형광성 면역검정(FIA), 효소 면역검정(EIA), 혼탁 억제 면역검정(nephelometric inhibition immunoassay: NIA), 효소 결합된 면역흡착 검정(ELISA), 및 방사면역검정(RIA)을 포함한다. 지시인자 모이어티, 또는 표지 그룹을 대상체 항체에 부착시킬 수 있으며 검정 장치 및 혼용성 면역검정 과정의 이용가능성에 의해 흔히 해석되는 방법의 다양한 사용의 필요성을 충족시키기 위해 선택된다. 적절한 표지는 방사성 핵종(예를 들면 125I, 131I, 35S, 3H, 또는 32P), 효소(예를 들면, 알칼린 포스파타제, 서양고추냉이 퍼옥시다제, 루시퍼라제, 또는 p-갈락토시다제), 형광성 모이어티 또는 단백질(예를 들면, 플루오레세인, 로다민, 피코에리트린, GFP, 또는 BFP), 또는 발광성 모이어티(예를 들면, 캘리포니아주 팔로 알토에 소재하는 Quantum Dot Corporation에 의해 공급된 QdotTM 나노입자)를 포함하나, 이에 한정되지 않는다. 상기 나타낸 다양한 면역검정을 수행하는데 사용되는 일반적인 기술은 당해 분야의 통상의 기술자에게 알려져 있다.
진단 목적을 위해, IL13R/IL4R 연속된 폴리펩타이드를 검출가능한 모이어티, 예를 들면, 그러나 이에 한정되지 않는, 당해 분야에 공지된 방사선동위원소, 형광성 표지, 및 다양한 효소-기질 표지로 표지할 수 있다. 폴리펩타이드에 대해 표지를 접합시키는 방법은 당해 분야에 공지되어 있다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 표지될 필요가 없으며, 이의 존재는 예를 들면, IL13R/IL4R 연속된 폴리펩타이드에 결합하는 항체를 사용하여 검출될 수 있다. 일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 임의의 공지된 검정 방법, 예를 들면, 경쟁적 결합 검정, 직접 및 간접 샌드위치 검정, 및 면역침전 검정에서 사용될 수 있다. Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987). 항-IL13 및 IL4 항체 및 폴리펩타이드는 또한 생체내 진단 검정, 예를 들면, 생체내 영상화에 사용될 수 있다. 일반적으로, 항체 또는 폴리펩타이드는 방사성 핵종(예를 들면, 111In, 99Tc, 14C, 131I, 125I, 3H, 또는 임의의 다른 방사성 핵종, 예를 들면, 본원에 기술된 것)으로 표지함으로써 목적한 세포 또는 조직을 임뮤노신티오그래피(immunoscintiography)를 사용하여 국재화할 수 있다. IL13R/IL4R 연속된 폴리펩타이드는 또한 당해 분야에 잘 공지된 기술을 사용하여 병리학에서 염색 시약으로서 사용할 수 있다.
일부 구현예에서, IL13R/IL4R 연속된 폴리펩타이드는 진단제로서 사용되며 IL13R/IL4R 연속된 폴리펩타이드는 치료제로서 사용된다. 일부 구현예에서, 제1 및 제2의 IL13R/IL4R 연속된 폴리펩타이드는 상이하다.
다음의 실시예는 개시내용의 특수한 양태를 나타내며 임의의 방식으로 개시내용을 제한하는 것으로 의도되지 않는다.
실시예
실시예 1
개과 IL4 및 IL13의 발현 및 정제
개과 IL13 단백질을 암호화하는 뉴클레오타이드 서열(서열 번호: 4)을 C-말단 끝에서 폴리-His 태그(poly-His tag)로 합성하여 포유동물 발현 벡터내로 클로닝하고 293 세포 또는 CHOS로 형질감염시켰다. 동일한 방법을 사용하여 C-말단 끝에 폴리-His 태그를 지닌 개과 IL4 단백질(서열 번호: 1)을 암호화하는 뉴클레오타이드를 발현시켰다.
개과 IL13 단백질을 함유하는 상층액을 수집하여 여과하였다. 개과 IL13은 Ni-NTA 컬럼(CaptivA® 단백질 A 친화성 수지, Repligen)을 사용하여 친화성 정제하였다. 동일한 방법을 사용하여 개과 IL4를 정제하였다.
실시예 2
IL13R 및 IL4R의 세포외 도메인
개과, 고양이과 및 말과 IL4 및/또는 IL13을 결합시키는데 관여하는 개과, 고양이과, 및 말과 IL4R의 세포외 도메인을 확인하고 경계부를 정의하였다. 개과 IL4R, 고양이과 IL4R, 및 말과 IL4의 완전한 길이의 세포외 도메인을 각각 서열 번호: 23, 서열 번호: 25, 및 서열 번호: 27로 확인하였다. 생물학적 활성을 보유하도록 상정한 개과 IL4R, 고양이과 IL4R, 및 말과 IL4R의 세포외 도메인 단편을 각각 서열 번호: 33, 서열 번호: 35, 및 서열 번호: 37로 확인하였다.
개과, 고양이과, 및 말과 IL4 및/또는 IL13을 결합시키는데 관여하는 개과, 고양이과, 및 말과 IL13R의 세포외 도메인을 확인하여 경계부를 확인하였다. 개과 IL13R, 고양이과 IL13R, 및 말과 IL13R의 완전한 길이의 세포외 도메인을 각각 서열 번호: 22, 서열 번호: 24, 및 서열 번호: 26으로서 확인하였다. 생물학적 활성을 보유하도록 상정된 개과 IL13R, 고양이과 IL13R, 및 말과 IL13R의 세포외 도메인 단편을 각각 서열 번호: 32, 서열 번호: 34, 및 서열 번호: 36으로서 확인하였다.
개과 IL13R(서열 번호: 22의 18번 위치에서), 고양이과 IL13R(서열 번호: 24의 18번 위치에서), 및 말과 IL13R(서열 번호: 26의 18번 위치에서) 내에서 쌍을 이루지 않은 시스테인(Cys)을 정보적으로 확인하고 3-D 모델링을 기반으로 포매된(embedded)(예측되지 않은) 바와 같이 측정하였다. 쌍을 이루지 않은 시스테인은 이황화물 결합을 형성할 것 같지 않으며 응집 경향성은 낮다. 따라서, 이러한 Cys 잔기의 부위-지시된 돌연변이유발은 도입되지 않았다.
실시예 3
CHO 세포로부터 개과 IL13R/IL4R 연속된 폴리펩타이드의 발현 및 정제
Fc IgGB에 연결된 개과 IL13R/IL4R 연속된 폴리펩타이드를 암호화하는 뉴클레오타이드 서열을 신호 서열로 설계하였다. 연속된 폴리펩타이드 "IL13R-IL4R-IgGB" (서열 번호: 20)의 경우, IL13R(서열 번호: 22)의 세포외 도메인은 IL4R(서열 번호: 23)의 세포외 도메인을 선행한다. 연속된 폴리펩타이드 IL4R-IL13R-IgGB (서열 번호: 21)의 경우, IL4R의 세포외 도메인은 IL13R의 세포외 도메인을 선행한다.
뉴클레오타이드 서열은 화학적으로 합성되어 CHO 숙주 세포내로 경질감염시키기에 적합한 발현 벡터내로 삽입되었다. CHO 세포내로 형질감염 후, 융합 단백질은 세포로부터 분비되었다. 예를 들면, 융합 단백질은 단일 단계 단백질 A 컬럼 크로마토그래피로 정제하였다.
각각의 IL13R-IL4R-IgGB 및 IL4R-IL13R-IgGB을 발현시키고 단백질 A 컬럼 또는 다른 크로마토그래피 방법, 예를 들면, 이온 교환 컬럼 크로마토그래피, 소수성 상호작용 컬럼 크로마토그래피, 혼합된 방식 컬럼 크로마토그래피, 예를 들면, CHT, 또는 다중모드 양식 컬럼 크로마토그래피, 예를 들면, CaptoMMC를 사용하여 단일 단계로 정제할 수 있다. 저 pH 또는 다른 바이러스 불활성화 및 바이러스 제거 단계를 적용할 수 있다. 정제된 단백질은 부형제와 혼합될 수 있으며, 여과로 멸균시켜 본 발명의 약제학적 조성물을 제조할 수 있다. 약제학적 조성물은 아토피성 피부염 또는 천식을 지닌 개에게 IL13 및/또는 IL4에 결합하고/하거나 억제하기에 충분한 양으로 투여될 수 있다.
벡터를 이후에 사용하여 FreestyleMaxTM 형질감염 시약(Life Technologies)을 사용하여 CHO-S 세포내에서 파일러트-규모(pilot-scale) 형질감염을 수행하였다 상층액을 조건화된 배지를 투명하게 함으로써 수거하였다. 단백질을 단일 통과 단백질 A 크로마토그래피 단계로 정제하고 추가의 시험을 위해 사용하였다.
실시예 4
IL13 및 IL4 결합 활성의 증명
본 실시예는 IL13R-IL4R-IgGB(서열 번호:20) 및 IL4R-IL13R-IgGB(서열 번호:21) 둘 다가 개과 IL4 및 IL13에 치료학적 활성에 필수적인 역학으로 결합함을 입증한다.
결합 분석을 다음과 같이 바이오센서 옥테트(biosensor Octet)를 사용하여 수행하였다. 요약하면, 개과 IL4(293 세포를 사용하여 생산함)를 바이오티닐화하였다. 유리된 반응하지 않은 바이오틴을 연장성 투석에 의해 바이오티닐화된 IL4로부터 제거하였다. 바이오티닐화된 개과 IL4를 스트렙타비딘 센서 팁에서 포획하였다. 다양한 농도(12, 16, 및 44 nM)의 IL13R-IL4R-IgGB(서열 번호:20)과의 IL4 결합을 90초 동안 모니터링하였다. 해리는 600초 동안 모니터링하였다. 완충액만의 블랭크 곡선을 감하여 임의의 이동(drift)에 대해 교정하였다. 데이타를 ForteBioTM 데이타 분석 소프트웨어를 사용하여 1:1 결합 모델에 대해 조정함으로써 kon, koff, 및 Kd를 측정하였다. 희석 및 모든 결합 단계에 대한 완충액은 다음과 같았다: 20 mM 포스페이트, 150 mM NaCl, pH 7.2. IL13R-IL4R-IgGB 및 리간드 IL4에 대한 Kd는 8 x10-11이었다.
다양한 농도(40.7, 및 140 nM)의 IL4R-IL13R-IgGB(서열 번호:21)와의 개과 IL4 연합을 90초 동안 모니터링하였다. 해리는 600초 동안 모니터링하였다. 완충액만의 블랭크 곡선을 감하여 임의의 이동에 대해 교정하였다. 데이타를 ForteBioTM 데이타 분석 소프트웨어를 사용하여 1:1 결합 모델에 대해 조정함으로써 kon, koff, 및 Kd를 측정하였다. 희석 및 모든 결합 단계에 대한 완충액은 다음과 같았다: 20 mM 포스페이트, 150 mM NaCl, pH 7.2. IL14R-IL13R-IgGB 및 리간드 IL4에 대한 Kd는 1.1 x10-11이었다.
C-말단 폴리His 태그를 지닌 개과 IL4 및 개과 IL13을 발현시키고 293개 세포로부터 정제하였다. EZ-연결(Link) NHS-LC-바이오틴을 Thermo Scientific(제품 번호 #21336)로부터 구입하고, 스트렙타비딘 바이오센서는 ForteBio(제품 번호 #18-509)로부터 구입하였다.
IL13R-IL4R-IgGB(서열 번호:20)과의 IL4 및 IL13 순차적인 결합 실험을 수행하였다. 바이오티닐화된 개과 IL13R-IL4R-IgGB를 스트렙타디빈 센서 팁 위에서 포획하였다. 개과 IL13R-IL4R-IgGB를 (1) 개과 IL4에 이은 IL13 또는 (2) 개과 IL13에 이은 IL4에 PBS중 30μg/mL의 IL4 및 IL13의 농도를 사용하여 노출시켰다(도 1). 실험은 일단 IL13R-IL4R-IgGB가 IL13에 결합하면, 이것이 IL4에 결합하지 않을 수 있으며, 일단 IL4에 결합하면, IL13에 결합하는 이의 능력이 감소됨을 입증하였다.
IL4R-IL13R-IgGB(서열 번호:21)를 사용한 IL4 및 IL13 순차적인 결합 실험을 수행하였다. 바이오티닐화된 개과 IL4R-IL13R-IgGB를 스트렙타비딘 센서 팁 상에 포획하였다. 개과 IL4R-IL13R-IgGB를 (1) 개과 IL4에 이은 IL13 또는 (2) 개과 IL13에 이은 IL4에 PBS중 30μg/mL의 IL4 및 IL13의 농도를 사용하여 노출시켰다(도 2). 이러한 실험은 일단 IL4R-IL13R-IgGB가 IL13에 결합하면, 이것이 IL4에 결합하지 않을 수 있으며, 일단 IL4에 결합하면, IL13에 결합하는 이의 능력이 감소됨을 입증하였다.
IL4 또는 IL13에 대한 IL13R-IL4R-IgGB 및 IL4R-IL13R-IgGB의 강력한 결합은 IL4R 및 IL13R 둘 다에 의해 이루어진 동시 결합 기여에 기인한 것으로 고려된다.
실시예 5
개과 IL4R-IL13R-Fc(SINK)의 세포 기능적 활성
TF1 세포(ATCC 제품 번호 CRL-2003), 세포 표면 상에 내인성 인터루킨 4 수용체를 발현하는 사람 적백혈구 세포주(Erythroleukemic cell line)를 증식 검정에 사용하였다. 열 불활성화된, 10% 태아 송아지 혈청(Sigma, 제품 번호 2868) 및 대수 성장 상의 2 nM/ml 사람 GM-CSF(R&D System, 제품 번호 215-GM-010)가 보충된 RPMI1640(Gibco, 제품 번호 11875)에서 성장된 세포를 검정에 사용하였다. 세포를 PBS로 2회 세척하고 GM-CSF가 들어있지 않는 상기 배지에 재현탁시켰다. 웰당 20,000개의 세포를 96-웰 플레이트(Corning, 제품 번호 3610)에 플레이팅하였다. 개과 IL4R-IL13R-Fc(SINK)를 일련의 희석물에 가한 후 개과 IL4(Sino Biological Inc, 제품 번호 70021-DNAE-5)를 50 ng/ml에서 가하였다. 세포를 37℃, 5% CO2에서 48시간 동안 총 100 μl의 용적으로 항온처리하였다. 항온처리 말기에, 세포를 실온에서 냉각시키고 CellTiter-Glo® 발광성 세포 생존능 검정(Luminescent Cell Viability Assay)(Promega, 제품 번호 G7570)을 사용하여 세포 ATP 함량을 측정함으로써 증식/다양성에 대해 검정하였다.
당해 검정에서, 100 μl의 예비혼합된 시약 A 및 B를 각각의 웰에 가하였다. 오비탈 진탕기 상에서 2분 동안 진탕시킨 후에, 세포를 분해하였다. 루시페린의 일-산화를 세포 속에 존재하는 Mg2+ 및 ATP의 존재하에서 루시퍼라제로 촉매시켜서, 세포내에서 ATP의 양에 비례하는 발광성 신호를 생성시켰다. ATP의 양은 배양물 속에 존재하는 세포의 수에 직접적으로 비례한다. 플레이트를 실온에서 10분 동안 항온처리함으로써 발광성 신호를 안정화시키고 발광성을 Synergy HT 미세플레이트 판독기(Biotek,버몬트주 위누스키 소재)를 사용하여 검출하였다.
데이타를 4 매개변수 로지스틱 핏(logistic fit)을 사용하여 분석하였으며 IC50은 2.0 nM이다. 도 3을 참고한다.
SEQUENCE LISTING
<110> KINDRED BIOSCIENCES, INC.
<120> IL4/IL13 RECEPTOR MOLECULE FOR VETERINARY USE
<130> IPA191297-US
<150> US 62/488,509
<151> 2017-04-21
<160> 49
<170> PatentIn version 3.5
<210> 1
<211> 131
<212> PRT
<213> Canis lupus
<400> 1
Met Gly Leu Thr Ser Gln Leu Ile Pro Thr Leu Val Cys Leu Leu Ala
1 5 10 15
Leu Thr Ser Thr Phe Val His Gly His Asn Phe Asn Ile Thr Ile Lys
20 25 30
Glu Ile Ile Lys Met Leu Asn Ile Leu Thr Ala Arg Asn Asp Ser Cys
35 40 45
Met Glu Leu Thr Val Asp Val Phe Thr Ala Pro Lys Asn Thr Ser Asp
50 55 60
Lys Glu Ile Phe Cys Arg Ala Ala Thr Val Leu Arg Gln Ile Tyr Thr
65 70 75 80
His Asn Cys Ser Asn Arg Tyr Leu Arg Gly Leu Tyr Arg Asn Leu Ser
85 90 95
Ser Met Ala Asn Lys Thr Cys Ser Met Asn Glu Ile Lys Lys Ser Thr
100 105 110
Leu Lys Asp Phe Leu Glu Arg Leu Lys Val Ile Met Gln Lys Lys Tyr
115 120 125
Tyr Arg His
130
<210> 2
<211> 132
<212> PRT
<213> Felis catus
<400> 2
Met Asp Leu Thr Ser Gln Leu Ile Pro Ala Leu Val Cys Leu Leu Ala
1 5 10 15
Phe Thr Ser Thr Phe Val His Gly Gln Asn Phe Asn Asn Thr Leu Lys
20 25 30
Glu Ile Ile Lys Thr Leu Asn Ile Leu Thr Ala Arg Asn Asp Ser Cys
35 40 45
Met Glu Leu Thr Met Asp Val Leu Ala Ala Pro Lys Asn Thr Ser Asp
50 55 60
Lys Glu Ile Phe Cys Arg Ala Thr Thr Val Leu Arg Gln Ile Tyr Thr
65 70 75 80
His His Asn Cys Ser Thr Lys Phe Leu Lys Gly Leu Asp Arg Asn Leu
85 90 95
Ser Ser Met Ala Asn Arg Thr Cys Ser Val Asn Glu Val Lys Lys Cys
100 105 110
Thr Leu Lys Asp Phe Leu Glu Arg Leu Lys Ala Ile Met Gln Lys Lys
115 120 125
Tyr Ser Lys His
130
<210> 3
<211> 137
<212> PRT
<213> Equus caballus
<400> 3
Met Gly Leu Thr Tyr Gln Leu Ile Pro Ala Leu Val Cys Leu Leu Ala
1 5 10 15
Cys Thr Ser Asn Phe Ile Gln Gly Cys Lys Tyr Asp Ile Thr Leu Gln
20 25 30
Glu Ile Ile Lys Thr Leu Asn Asn Leu Thr Asp Gly Lys Gly Lys Asn
35 40 45
Ser Cys Met Glu Leu Thr Val Ala Asp Ala Phe Ala Gly Pro Lys Asn
50 55 60
Thr Asp Gly Lys Glu Ile Cys Arg Ala Ala Lys Val Leu Gln Gln Leu
65 70 75 80
Tyr Lys Arg His Asp Arg Ser Leu Ile Lys Glu Cys Leu Ser Gly Leu
85 90 95
Asp Arg Asn Leu Lys Gly Met Ala Asn Gly Thr Cys Cys Thr Val Asn
100 105 110
Glu Ala Lys Lys Ser Thr Leu Lys Asp Phe Leu Glu Arg Leu Lys Thr
115 120 125
Ile Met Lys Glu Lys Tyr Ser Lys Cys
130 135
<210> 4
<211> 131
<212> PRT
<213> Canis lupus
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Met Ala Leu Trp Leu Thr Val Val Ile Ala Leu Thr Cys Leu Gly Gly
1 5 10 15
Leu Ala Ser Pro Ser Pro Val Thr Pro Ser Pro Thr Leu Lys Glu Leu
20 25 30
Ile Glu Glu Leu Val Asn Ile Thr Gln Asn Gln Ala Ser Leu Cys Asn
35 40 45
Gly Ser Met Val Trp Ser Val Asn Leu Thr Ala Gly Met Tyr Cys Ala
50 55 60
Ala Leu Glu Ser Leu Ile Asn Val Ser Asp Cys Ser Ala Ile Gln Arg
65 70 75 80
Thr Gln Arg Met Leu Lys Ala Leu Cys Ser Gln Lys Pro Ala Ala Gly
85 90 95
Gln Ile Ser Ser Glu Arg Ser Arg Asp Thr Lys Ile Glu Val Ile Gln
100 105 110
Leu Val Lys Asn Leu Leu Thr Tyr Val Arg Gly Val Tyr Arg His Gly
115 120 125
Asn Phe Arg
130
<210> 5
<211> 183
<212> PRT
<213> Felis catus
<400> 5
Met Trp Phe Leu Asp Ser Thr Arg Gln Ser Gly Asp Gln Gly Gly Arg
1 5 10 15
Arg His Thr Trp Pro Ile Lys Ala Thr Ala Arg Gly Gln Gly His Lys
20 25 30
Pro Leu Ser Leu Gly Gln Pro Thr Cys Pro Leu Leu Ala Pro Pro Val
35 40 45
Leu Ala Leu Gly Ser Met Ala Leu Trp Leu Thr Val Val Ile Ala Leu
50 55 60
Thr Cys Leu Gly Gly Leu Ala Ser Pro Gly Pro His Ser Arg Arg Glu
65 70 75 80
Leu Lys Glu Leu Ile Glu Glu Leu Val Asn Ile Thr Gln Asn Gln Val
85 90 95
Ser Leu Cys Asn Gly Ser Met Val Trp Ser Val Asn Leu Thr Thr Gly
100 105 110
Met Tyr Cys Ala Ala Leu Glu Ser Leu Ile Asn Val Ser Asp Cys Thr
115 120 125
Ala Ile Gln Arg Thr Gln Arg Met Leu Lys Ala Leu Cys Thr Gln Lys
130 135 140
Pro Ser Ala Gly Gln Thr Ala Ser Glu Arg Ser Arg Asp Thr Lys Ile
145 150 155 160
Glu Val Ile Gln Leu Val Lys Asn Leu Leu Asn His Leu Arg Arg Asn
165 170 175
Phe Arg His Gly Asn Phe Lys
180
<210> 6
<211> 133
<212> PRT
<213> Equus caballus
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Met Ala Leu Trp Leu Thr Ala Val Ile Ala Leu Ala Cys Leu Gly Gly
1 5 10 15
Leu Ala Ser Pro Ala Pro Leu Pro Ser Ser Met Ala Leu Lys Glu Leu
20 25 30
Ile Lys Glu Leu Val Asn Ile Thr Gln Asn Gln Ala Pro Leu Cys Asn
35 40 45
Gly Ser Met Val Trp Ser Val Asn Leu Thr Ala Asp Thr Tyr Cys Arg
50 55 60
Ala Leu Glu Ser Leu Ser Asn Val Ser Thr Cys Ser Ala Ile Gln Asn
65 70 75 80
Thr Arg Lys Met Leu Thr Lys Leu Cys Pro His Gln Leu Ser Ala Gly
85 90 95
Gln Val Ser Ser Glu Arg Ala Arg Asp Thr Lys Ile Glu Val Ile Val
100 105 110
Leu Val Lys Asp Leu Leu Lys Asn Leu Arg Lys Ile Phe His Gly Gly
115 120 125
Lys His Val Asp Ala
130
<210> 7
<211> 823
<212> PRT
<213> Canis lupus
<400> 7
Met Gly Arg Leu Cys Ser Gly Leu Thr Phe Pro Val Ser Cys Leu Val
1 5 10 15
Leu Val Trp Val Ala Ser Ser Gly Ser Val Lys Val Leu His Glu Pro
20 25 30
Ser Cys Phe Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met
35 40 45
Asp His Pro Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu
50 55 60
Asp Phe Met Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu
65 70 75 80
Asp Ser Val Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala
85 90 95
Asp Val Tyr Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser
100 105 110
Gly Ser Phe Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn
115 120 125
Leu Thr Val His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr
130 135 140
Asn Pro Tyr Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met
145 150 155 160
Val Asn Val Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn
165 170 175
Val Thr Tyr Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys
180 185 190
Ser Gly Ala Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr
195 200 205
Asn Ser Thr Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr
210 215 220
Tyr Glu Pro Trp Glu Gln His Leu Pro Leu Gly Val Ser Ile Ser Cys
225 230 235 240
Leu Val Ile Leu Ala Ile Cys Leu Ser Cys Tyr Phe Ser Ile Ile Lys
245 250 255
Ile Lys Lys Gly Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser Pro
260 265 270
Leu Val Ala Ile Val Ile Gln Asp Ser Gln Val Ser Leu Trp Gly Lys
275 280 285
Arg Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro His Trp Lys Thr Cys
290 295 300
Leu Thr Lys Leu Leu Pro Cys Leu Leu Glu His Gly Leu Gly Arg Glu
305 310 315 320
Glu Glu Ser Pro Lys Thr Ala Lys Asn Gly Pro Leu Gln Gly Pro Gly
325 330 335
Lys Pro Ala Trp Cys Pro Val Glu Val Ser Lys Thr Ile Leu Trp Pro
340 345 350
Glu Ser Ile Ser Val Val Gln Cys Val Glu Leu Ser Glu Ala Pro Val
355 360 365
Asp Asn Glu Glu Glu Glu Glu Val Glu Glu Asp Lys Arg Ser Leu Cys
370 375 380
Pro Ser Leu Glu Gly Ser Gly Gly Ser Phe Gln Glu Gly Arg Glu Gly
385 390 395 400
Ile Val Ala Arg Leu Thr Glu Ser Leu Phe Leu Asp Leu Leu Gly Gly
405 410 415
Glu Asn Gly Gly Phe Cys Pro Gln Gly Leu Glu Glu Ser Cys Leu Pro
420 425 430
Pro Pro Ser Gly Ser Val Gly Ala Gln Met Pro Trp Ala Gln Phe Pro
435 440 445
Arg Ala Gly Pro Arg Ala Ala Pro Glu Gly Pro Glu Gln Pro Arg Arg
450 455 460
Pro Glu Ser Ala Leu Gln Ala Ser Pro Thr Gln Ser Ala Gly Ser Ser
465 470 475 480
Ala Phe Pro Glu Pro Pro Pro Val Val Thr Asp Asn Pro Ala Tyr Arg
485 490 495
Ser Phe Gly Ser Phe Leu Gly Gln Ser Ser Asp Pro Gly Asp Gly Asp
500 505 510
Ser Asp Pro Glu Leu Ala Asp Arg Pro Gly Glu Ala Asp Pro Gly Ile
515 520 525
Pro Ser Ala Pro Gln Pro Pro Glu Pro Pro Ala Ala Leu Gln Pro Glu
530 535 540
Pro Glu Ser Trp Glu Gln Ile Leu Arg Gln Ser Val Leu Gln His Arg
545 550 555 560
Ala Ala Pro Ala Pro Gly Pro Gly Pro Gly Ser Gly Tyr Arg Glu Phe
565 570 575
Thr Cys Ala Val Lys Gln Gly Ser Ala Pro Asp Ala Gly Gly Pro Gly
580 585 590
Phe Gly Pro Ser Gly Glu Ala Gly Tyr Lys Ala Phe Cys Ser Leu Leu
595 600 605
Pro Gly Gly Ala Thr Cys Pro Gly Thr Ser Gly Gly Glu Ala Gly Ser
610 615 620
Gly Glu Gly Gly Tyr Lys Pro Phe Gln Ser Leu Thr Pro Gly Cys Pro
625 630 635 640
Gly Ala Pro Thr Pro Val Pro Val Pro Leu Phe Thr Phe Gly Leu Asp
645 650 655
Thr Glu Pro Pro Gly Ser Pro Gln Asp Ser Leu Gly Ala Gly Ser Ser
660 665 670
Pro Glu His Leu Gly Val Glu Pro Ala Gly Lys Glu Glu Asp Ser Arg
675 680 685
Lys Thr Leu Leu Ala Pro Glu Gln Ala Thr Asp Pro Leu Arg Asp Asp
690 695 700
Leu Ala Ser Ser Ile Val Tyr Ser Ala Leu Thr Cys His Leu Cys Gly
705 710 715 720
His Leu Lys Gln Trp His Asp Gln Glu Glu Arg Gly Lys Ala His Ile
725 730 735
Val Pro Ser Pro Cys Cys Gly Cys Cys Cys Gly Asp Arg Ser Ser Leu
740 745 750
Leu Leu Ser Pro Leu Arg Ala Pro Asn Val Leu Pro Gly Gly Val Leu
755 760 765
Leu Glu Ala Ser Leu Ser Pro Ala Ser Leu Val Pro Ser Gly Val Ser
770 775 780
Lys Glu Gly Lys Ser Ser Pro Phe Ser Gln Pro Ala Ser Ser Ser Ala
785 790 795 800
Gln Ser Ser Ser Gln Thr Pro Lys Lys Leu Ala Val Leu Ser Thr Glu
805 810 815
Pro Thr Cys Met Ser Ala Ser
820
<210> 8
<211> 881
<212> PRT
<213> Felis catus
<400> 8
Met Gly Arg Leu Cys Ser Gly Leu Thr Phe Pro Val Ser Cys Leu Ile
1 5 10 15
Leu Met Trp Ala Ala Gly Ser Gly Ser Val Lys Val Leu Arg Ala Pro
20 25 30
Thr Cys Phe Ser Asp Tyr Phe Ser Thr Ser Val Cys Gln Trp Asn Met
35 40 45
Asp Ala Pro Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu
50 55 60
Asn Phe Met Gly Ser Glu Asn Arg Thr Cys Val Pro Glu Asn Gly Glu
65 70 75 80
Gly Ala Ala Cys Ala Cys Ser Met Leu Met Asp Asp Phe Val Glu Ala
85 90 95
Asp Val Tyr Gln Leu His Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser
100 105 110
Gly Ser Phe Lys Pro Ser Ser His Val Lys Pro Arg Ala Pro Gly Asn
115 120 125
Leu Thr Val His Pro Asn Val Ser His Thr Trp Leu Leu Arg Trp Ser
130 135 140
Asn Pro Tyr Pro Pro Glu Asn His Leu His Ala Glu Leu Thr Tyr Met
145 150 155 160
Val Asn Ile Ser Ser Glu Asp Asp Pro Thr Asp Val Ser Val Cys Ala
165 170 175
Ser Gly Phe Leu Cys His Leu Leu Gly Leu Arg Arg Val Glu Thr Gly
180 185 190
Ala Pro Gly Ala Arg Leu Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg
195 200 205
Arg Val Pro Gly Ser Gln Cys Ala Val Ile Ser Cys Cys Arg Trp Val
210 215 220
Leu Ile Ala Leu Thr Ser Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly
225 230 235 240
Leu Arg Ser Gln Thr Arg Tyr Val Ser Val Ala Glu Gly Leu Phe Gly
245 250 255
Ala Thr Pro Arg Val Leu Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser
260 265 270
Ala Ala Arg Glu Gln Met Ser Pro Asp Pro Ser Ala Phe His Ser Ile
275 280 285
Asp Tyr Glu Pro Trp Glu Gln His Leu Pro Leu Gly Val Ser Ile Ser
290 295 300
Cys Leu Val Ile Leu Ala Val Cys Leu Ser Cys Tyr Leu Ser Val Ile
305 310 315 320
Lys Ile Lys Lys Glu Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser
325 330 335
His Leu Val Ala Ile Val Ile Gln Asp Pro Gln Val Ser Leu Trp Gly
340 345 350
Lys Arg Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro His Trp Lys Thr
355 360 365
Cys Leu Arg Lys Leu Leu Pro Cys Leu Leu Glu His Gly Met Glu Arg
370 375 380
Lys Glu Asp Pro Ser Lys Ile Ala Arg Asn Gly Pro Ser Gln Cys Ser
385 390 395 400
Gly Lys Ser Ala Trp Cys Pro Val Glu Val Ser Lys Thr Ile Leu Trp
405 410 415
Pro Glu Ser Ile Ser Val Val Arg Cys Val Glu Leu Leu Glu Ala Pro
420 425 430
Val Glu Ser Glu Glu Glu Glu Glu Glu Glu Glu Asp Lys Gly Ser Phe
435 440 445
Cys Pro Ser Pro Val Asn Leu Glu Asp Ser Phe Gln Glu Gly Arg Glu
450 455 460
Gly Ile Ala Ala Arg Leu Thr Glu Ser Leu Phe Met Asp Leu Leu Gly
465 470 475 480
Val Glu Lys Gly Gly Phe Gly Pro Gln Gly Ser Leu Glu Ser Trp Phe
485 490 495
Pro Pro Pro Ser Gly Ser Ala Gly Ala Gln Met Pro Trp Ala Glu Phe
500 505 510
Pro Gly Pro Gly Pro Gln Glu Ala Ser Pro Gln Gly Lys Glu Gln Pro
515 520 525
Phe Asp Pro Arg Ser Asp Pro Leu Ala Thr Leu Pro Gln Ser Pro Ala
530 535 540
Ser Pro Thr Phe Pro Glu Thr Pro Pro Val Val Thr Asp Asn Pro Ala
545 550 555 560
Tyr Arg Ser Phe Gly Thr Phe Gln Gly Arg Ser Ser Gly Pro Gly Glu
565 570 575
Cys Asp Ser Gly Pro Glu Leu Ala Gly Arg Leu Gly Glu Ala Asp Pro
580 585 590
Gly Ile Pro Ala Ala Pro Gln Pro Ser Glu Pro Pro Ser Ala Leu Gln
595 600 605
Pro Glu Ala Glu Thr Trp Glu Gln Ile Leu Arg Gln Arg Val Leu Gln
610 615 620
His Arg Gly Ala Pro Ala Pro Ala Pro Gly Ser Gly Tyr Arg Glu Phe
625 630 635 640
Val Cys Ala Val Arg Gln Gly Ser Thr Gln Asp Ser Gly Val Gly Asp
645 650 655
Phe Gly Pro Ser Glu Glu Ala Gly Tyr Lys Ala Phe Ser Ser Leu Leu
660 665 670
Thr Ser Gly Ala Val Cys Pro Glu Ser Gly Gly Glu Ala Gly Ser Gly
675 680 685
Asp Gly Gly Tyr Lys Pro Phe Gln Ser Leu Thr Pro Gly Cys Pro Gly
690 695 700
Ala Pro Ala Pro Val Pro Val Pro Leu Phe Thr Phe Gly Leu Asp Ala
705 710 715 720
Glu Pro Pro His Cys Pro Gln Asp Ser Pro Leu Pro Gly Ser Ser Pro
725 730 735
Glu Pro Ala Gly Lys Ala Gln Asp Ser His Lys Thr Pro Pro Ala Pro
740 745 750
Glu Gln Ala Ala Asp Pro Leu Arg Asp Asp Leu Ala Ser Gly Ile Val
755 760 765
Tyr Ser Ala Leu Thr Cys His Leu Cys Gly His Leu Lys Gln Cys His
770 775 780
Gly Gln Glu Glu Gly Gly Glu Ala His Pro Val Ala Ser Pro Cys Cys
785 790 795 800
Gly Cys Cys Cys Gly Asp Arg Ser Ser Pro Leu Val Ser Pro Leu Arg
805 810 815
Ala Pro Asp Pro Leu Pro Gly Gly Val Pro Leu Glu Ala Ser Leu Ser
820 825 830
Pro Ala Ser Pro Ala Pro Leu Ala Val Ser Glu Glu Gly Pro Pro Ser
835 840 845
Leu Cys Phe Gln Pro Ala Leu Ser His Ala His Ser Ser Ser Gln Thr
850 855 860
Pro Lys Lys Val Ala Met Leu Ser Pro Glu Pro Thr Cys Thr Met Ala
865 870 875 880
Ser
<210> 9
<211> 809
<212> PRT
<213> Equus caballus
<400> 9
Met Gly Cys Leu Cys Pro Gly Leu Thr Leu Pro Val Ser Cys Leu Ile
1 5 10 15
Leu Val Trp Ala Ala Gly Ser Gly Ser Val Lys Val Leu His Leu Thr
20 25 30
Ala Cys Phe Ser Asp Tyr Ile Ser Ala Ser Thr Cys Glu Trp Lys Met
35 40 45
Asp Arg Pro Thr Asn Cys Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu
50 55 60
Asn Asp Glu Phe Ser Asp Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu
65 70 75 80
Asp Glu Val Cys Val Cys Arg Met Leu Met Asp Asn Ile Val Ser Glu
85 90 95
Asp Val Tyr Glu Leu Asp Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn
100 105 110
Ser Ser Phe Lys Pro Ser Arg His Val Lys Pro Arg Ala Pro Gln Asn
115 120 125
Leu Thr Val His Ala Ile Ser His Thr Trp Leu Leu Thr Trp Ser Asn
130 135 140
Pro Tyr Pro Leu Lys Asn His Leu Trp Ser Glu Leu Thr Tyr Leu Val
145 150 155 160
Asn Ile Ser Lys Glu Asp Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val
165 170 175
Thr Tyr Met Asp Pro Thr Leu Arg Val Thr Ala Ser Thr Leu Lys Ser
180 185 190
Arg Ala Thr Tyr Ser Ala Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn
195 200 205
Ser Thr Trp Ser Glu Trp Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr
210 215 220
Glu Gln Pro Leu Glu Gln Arg Leu Pro Leu Gly Val Ser Ile Ser Cys
225 230 235 240
Val Val Ile Leu Ala Ile Cys Leu Ser Cys Tyr Phe Ser Ile Ile Lys
245 250 255
Ile Lys Lys Glu Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser Pro
260 265 270
Leu Val Ala Ile Val Leu Gln Asp Ser Gln Val Ser Leu Trp Gly Lys
275 280 285
Gln Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro Arg Trp Lys Thr Cys
290 295 300
Leu Thr Lys Leu Leu Pro Cys Leu Leu Glu His Gly Leu Gln Lys Glu
305 310 315 320
Glu Asp Ser Ser Lys Thr Val Arg Asn Gly Pro Phe Gln Ser Pro Gly
325 330 335
Lys Ser Ala Trp His Thr Val Glu Val Asn His Thr Ile Leu Arg Pro
340 345 350
Glu Ile Ile Ser Val Val Pro Cys Val Glu Leu Cys Glu Ala Gln Val
355 360 365
Glu Ser Glu Glu Glu Glu Val Glu Glu Asp Arg Gly Ser Phe Cys Pro
370 375 380
Ser Pro Glu Ser Ser Gly Ser Gly Phe Gln Glu Gly Arg Glu Gly Val
385 390 395 400
Ala Ala Arg Leu Thr Glu Ser Leu Phe Leu Gly Leu Leu Gly Ala Glu
405 410 415
Asn Gly Ala Leu Gly Glu Ser Cys Leu Leu Pro Pro Leu Gly Ser Ala
420 425 430
His Met Pro Trp Ala Arg Ile Ser Ser Ala Gly Pro Gln Glu Ala Ala
435 440 445
Ser Gln Gly Glu Glu Gln Pro Leu Asn Pro Glu Ser Asn Pro Leu Ala
450 455 460
Thr Leu Thr Gln Ser Pro Gly Ser Leu Ala Phe Thr Glu Ala Pro Ala
465 470 475 480
Val Val Ala Asp Asn Pro Ala Tyr Arg Ser Phe Ser Asn Ser Leu Ser
485 490 495
Gln Pro Arg Gly Pro Gly Glu Leu Asp Ser Asp Pro Gln Leu Ala Glu
500 505 510
His Leu Gly Gln Val Asp Pro Ser Ile Pro Ser Ala Pro Gln Pro Ser
515 520 525
Glu Pro Pro Thr Ala Leu Gln Pro Glu Pro Glu Thr Trp Glu Gln Met
530 535 540
Leu Arg Gln Ser Val Leu Gln Gln Gly Ala Ala Pro Ala Pro Ala Ser
545 550 555 560
Ala Pro Thr Gly Gly Tyr Arg Glu Phe Ala Gln Ala Val Lys Gln Gly
565 570 575
Gly Gly Ala Ala Gly Ser Gly Pro Ser Gly Glu Ala Gly Tyr Lys Ala
580 585 590
Phe Ser Ser Leu Leu Ala Gly Ser Ala Val Cys Pro Gly Gln Ser Gly
595 600 605
Val Glu Ala Ser Ser Gly Glu Gly Gly Tyr Arg Pro Tyr Glu Ser Pro
610 615 620
Asp Pro Gly Ala Pro Ala Pro Val Pro Val Pro Leu Phe Thr Phe Gly
625 630 635 640
Leu Asp Val Glu Pro Pro His Ser Pro Gln Asn Ser Leu Leu Pro Gly
645 650 655
Gly Ser Pro Glu Leu Pro Gly Pro Glu Pro Thr Val Lys Gly Glu Asp
660 665 670
Pro Arg Lys Pro Leu Leu Ser Ala Gln Gln Ala Thr Asp Ser Leu Arg
675 680 685
Asp Asp Leu Gly Ser Gly Ile Val Tyr Ser Ala Leu Thr Cys His Leu
690 695 700
Cys Gly His Leu Lys Gln Cys His Gly Gln Glu Glu His Gly Glu Ala
705 710 715 720
His Thr Val Ala Ser Pro Cys Cys Gly Cys Cys Cys Gly Asp Arg Ser
725 730 735
Ser Pro Pro Val Ser Pro Val Arg Ala Leu Asp Pro Pro Pro Gly Gly
740 745 750
Val Pro Leu Glu Ala Gly Leu Ser Leu Ala Ser Leu Gly Ser Leu Gly
755 760 765
Leu Ser Glu Glu Arg Lys Pro Ser Leu Phe Phe Gln Pro Ala Pro Gly
770 775 780
Asn Ala Gln Ser Ser Ser Gln Thr Pro Leu Thr Val Ala Met Leu Ser
785 790 795 800
Thr Gly Pro Thr Cys Thr Ser Ala Ser
805
<210> 10
<211> 427
<212> PRT
<213> Canis lupus
<400> 10
Met Glu Arg Pro Ala Arg Leu Cys Gly Leu Trp Ala Leu Leu Leu Cys
1 5 10 15
Ala Ala Gly Gly Arg Gly Gly Gly Val Ala Ala Pro Thr Glu Thr Gln
20 25 30
Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr Val
35 40 45
Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr Leu
50 55 60
Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala Pro
65 70 75 80
Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys Leu
85 90 95
Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser Ile
100 105 110
Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser Ala
115 120 125
Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys Cys
130 135 140
Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr Leu
145 150 155 160
Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp Ile
165 170 175
Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn Leu
180 185 190
Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys Asp
195 200 205
Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr Ser
210 215 220
His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln Asn
225 230 235 240
Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser Arg
245 250 255
Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr Asn
260 265 270
Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe Glu
275 280 285
Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu Pro
290 295 300
Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu Cys
305 310 315 320
Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser Ile
325 330 335
Gly Glu Asn Thr Asp Pro Thr Phe Tyr Ile Thr Met Leu Leu Ala Thr
340 345 350
Pro Val Ile Val Ala Gly Ala Ile Ile Val Leu Leu Leu Tyr Leu Lys
355 360 365
Arg Leu Lys Ile Ile Ile Phe Pro Pro Ile Pro Asp Pro Gly Lys Ile
370 375 380
Phe Lys Glu Met Phe Gly Asp Gln Asn Asp Asp Thr Leu His Trp Arg
385 390 395 400
Lys Tyr Asp Ile Tyr Glu Lys Gln Thr Lys Glu Glu Thr Asp Ser Val
405 410 415
Val Leu Ile Glu Asn Leu Lys Lys Ala Ser Gln
420 425
<210> 11
<211> 479
<212> PRT
<213> Felis catus
<400> 11
Met Met Thr Lys Cys Ser Ser Asp Arg Asn Val Phe Lys Arg Lys Trp
1 5 10 15
Phe Leu Phe Pro Ala Ser Gln Tyr Thr Phe Arg Pro Ile His Gln Ala
20 25 30
Arg Pro Cys Glu Val Pro Ala Val His Leu Glu Pro Ser Pro Pro Trp
35 40 45
Glu Val Gly Leu Gly Leu Leu Asn Leu Glu Ser Glu Phe Arg Lys Leu
50 55 60
Gly Leu Arg Gly Arg Arg Leu Ala Ala Ala Pro Pro Asp Ser Arg Ala
65 70 75 80
Glu Ala Ala Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser
85 90 95
Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly
100 105 110
Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys
115 120 125
Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro
130 135 140
Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn
145 150 155 160
Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro
165 170 175
Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His
180 185 190
Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser
195 200 205
Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys
210 215 220
Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys
225 230 235 240
Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser
245 250 255
Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe
260 265 270
Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile
275 280 285
Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn
290 295 300
Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn
305 310 315 320
Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys
325 330 335
Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe
340 345 350
Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg
355 360 365
Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn
370 375 380
Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Phe Tyr
385 390 395 400
Ile Thr Met Leu Leu Ala Thr Pro Val Ile Val Ala Gly Ala Ile Ile
405 410 415
Val Leu Leu Leu Tyr Leu Lys Arg Leu Lys Ile Ile Ile Phe Pro Pro
420 425 430
Ile Pro Asp Pro Gly Lys Ile Phe Lys Glu Met Phe Gly Asp Gln Asn
435 440 445
Asp Asp Ser Leu His Trp Lys Lys Tyr Asp Ile Tyr Glu Lys Gln Thr
450 455 460
Lys Glu Glu Thr Asp Ser Val Val Leu Ile Glu Asn Ala Ser Gln
465 470 475
<210> 12
<211> 407
<212> PRT
<213> Equus caballus
<400> 12
Met Tyr Phe Leu Cys Leu Ile Trp Thr Glu Ser Gln Pro Pro Val Thr
1 5 10 15
Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp
20 25 30
Asn Pro Pro Glu Gly Val Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser
35 40 45
His Phe Gly Asn Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg
50 55 60
Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser
65 70 75 80
Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys
85 90 95
Cys Ile Ser Pro Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu
100 105 110
Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro
115 120 125
Gly Lys Asn Ala Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His
130 135 140
Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly
145 150 155 160
Gln His Ile Gly Cys Ser Phe Ala Leu Thr Glu Val Lys Asp Ser Ile
165 170 175
Phe Glu Gln His Ser Val Gln Ile Met Val Lys Asp Asn Ala Gly Lys
180 185 190
Ile Arg Pro Phe Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro
195 200 205
Asp Pro Pro His Ile Lys Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr
210 215 220
Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr
225 230 235 240
Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser
245 250 255
Val Glu Glu Ala Lys Cys Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu
260 265 270
Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu Pro Asp Thr Val Asn
275 280 285
Thr Val Arg Ile Arg Val Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp
290 295 300
Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser Ile Gly Lys Lys Ala
305 310 315 320
Asp Pro Thr Phe Tyr Ile Ala Met Leu Leu Ile Ile Pro Val Ile Val
325 330 335
Ala Gly Ala Ile Ile Val Leu Leu Leu Tyr Leu Lys Arg Leu Lys Ile
340 345 350
Ile Met Phe Pro Pro Ile Pro Asp Pro Gly Lys Ile Phe Lys Glu Met
355 360 365
Phe Gly Asp Gln Asn Asp Asp Thr Leu His Trp Lys Lys Tyr Asp Ile
370 375 380
Tyr Glu Lys Gln Thr Lys Glu Glu Thr Asp Ser Val Val Leu Ile Glu
385 390 395 400
Asn Leu Lys Arg Ala Ser Gln
405
<210> 13
<211> 761
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL13R-IL4R-Fc (without signal sequence)
<400> 13
Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro
20 25 30
Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile
165 170 175
Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu
195 200 205
Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly Gly Ser Gly
305 310 315 320
Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr
325 330 335
Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys
340 345 350
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu
355 360 365
Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys
370 375 380
Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp
385 390 395 400
Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser
405 410 415
Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn
420 425 430
Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu
435 440 445
Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp
450 455 460
Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro
465 470 475 480
Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser
485 490 495
Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp
500 505 510
Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Lys Arg Glu
515 520 525
Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro
530 535 540
Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys
545 550 555 560
Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val
565 570 575
Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp
580 585 590
Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe
595 600 605
Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp
610 615 620
Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu
625 630 635 640
Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His
645 650 655
Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys
660 665 670
Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp
675 680 685
Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys
690 695 700
Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu
705 710 715 720
Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr
725 730 735
Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
740 745 750
Glu Ser Leu Ser His Ser Pro Gly Lys
755 760
<210> 14
<211> 758
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R-IL13R-IgGA (without signal sequence)
<400> 14
Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu
35 40 45
Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys
50 55 60
Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp
65 70 75 80
Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser
85 90 95
Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu
115 120 125
Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp
130 135 140
Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro
145 150 155 160
Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser
165 170 175
Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp
180 185 190
Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly
195 200 205
Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val
210 215 220
Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala
225 230 235 240
Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln
245 250 255
Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu
260 265 270
Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu
275 280 285
Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu
290 295 300
Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn
305 310 315 320
Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro
325 330 335
Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile
340 345 350
Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser
355 360 365
Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val
370 375 380
Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn
385 390 395 400
Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys
405 410 415
Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro
420 425 430
Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn
435 440 445
Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys
450 455 460
Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met
465 470 475 480
Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val
485 490 495
Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp
500 505 510
Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Phe Asn Glu
515 520 525
Cys Arg Cys Thr Asp Thr Pro Pro Cys Pro Val Pro Glu Pro Leu Gly
530 535 540
Gly Pro Ser Val Leu Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg
545 550 555 560
Ile Thr Arg Thr Pro Glu Val Thr Cys Val Val Leu Asp Leu Gly Arg
565 570 575
Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val
580 585 590
His Thr Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr
595 600 605
Arg Val Val Ser Val Leu Pro Ile Glu His Gln Asp Trp Leu Thr Gly
610 615 620
Lys Glu Phe Lys Cys Arg Val Asn His Ile Asp Leu Pro Ser Pro Ile
625 630 635 640
Glu Arg Thr Ile Ser Lys Ala Arg Gly Arg Ala His Lys Pro Ser Val
645 650 655
Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val
660 665 670
Ser Ile Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val
675 680 685
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg Lys His Arg Met
690 695 700
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
705 710 715 720
Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys
725 730 735
Ala Val Met His Glu Thr Leu Gln Asn His Tyr Thr Asp Leu Ser Leu
740 745 750
Ser His Ser Pro Gly Lys
755
<210> 15
<211> 762
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R-IL13R-IgGB (without signal sequence)
<400> 15
Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu
35 40 45
Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys
50 55 60
Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp
65 70 75 80
Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser
85 90 95
Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu
115 120 125
Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp
130 135 140
Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro
145 150 155 160
Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser
165 170 175
Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp
180 185 190
Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly
195 200 205
Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val
210 215 220
Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala
225 230 235 240
Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln
245 250 255
Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu
260 265 270
Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu
275 280 285
Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu
290 295 300
Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn
305 310 315 320
Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro
325 330 335
Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile
340 345 350
Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser
355 360 365
Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val
370 375 380
Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn
385 390 395 400
Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys
405 410 415
Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro
420 425 430
Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn
435 440 445
Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys
450 455 460
Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met
465 470 475 480
Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val
485 490 495
Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp
500 505 510
Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Pro Lys Arg
515 520 525
Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
530 535 540
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
545 550 555 560
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
565 570 575
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
580 585 590
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
595 600 605
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
610 615 620
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala
625 630 635 640
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
645 650 655
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
660 665 670
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
675 680 685
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
690 695 700
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
705 710 715 720
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
725 730 735
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
740 745 750
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
755 760
<210> 16
<211> 760
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R-IL13R-IgGC (without signal sequence)
<400> 16
Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu
35 40 45
Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys
50 55 60
Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp
65 70 75 80
Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser
85 90 95
Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu
115 120 125
Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp
130 135 140
Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro
145 150 155 160
Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser
165 170 175
Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp
180 185 190
Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly
195 200 205
Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val
210 215 220
Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala
225 230 235 240
Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln
245 250 255
Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu
260 265 270
Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu
275 280 285
Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu
290 295 300
Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn
305 310 315 320
Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro
325 330 335
Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile
340 345 350
Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser
355 360 365
Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val
370 375 380
Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn
385 390 395 400
Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys
405 410 415
Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro
420 425 430
Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn
435 440 445
Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys
450 455 460
Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met
465 470 475 480
Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val
485 490 495
Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp
500 505 510
Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Ala Lys Glu
515 520 525
Cys Glu Cys Lys Cys Asn Cys Asn Asn Cys Pro Cys Pro Gly Cys Gly
530 535 540
Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
545 550 555 560
Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys Val Val Val Asp
565 570 575
Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp Phe Val Asp Ser
580 585 590
Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu Glu Gln Ser Asn
595 600 605
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
610 615 620
Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn Lys Ala Leu Pro
625 630 635 640
Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly Gln Ala His Gln
645 650 655
Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Met Ser Lys Asn
660 665 670
Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe Pro Pro Glu Ile
675 680 685
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
690 695 700
Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
705 710 715 720
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
725 730 735
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Ile
740 745 750
Ser Leu Ser His Ser Pro Gly Lys
755 760
<210> 17
<211> 758
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R-IL13R-IgGD (without signal sequence)
<400> 17
Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu
35 40 45
Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys
50 55 60
Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp
65 70 75 80
Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser
85 90 95
Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu
115 120 125
Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp
130 135 140
Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro
145 150 155 160
Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser
165 170 175
Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp
180 185 190
Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly
195 200 205
Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val
210 215 220
Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala
225 230 235 240
Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln
245 250 255
Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu
260 265 270
Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu
275 280 285
Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu
290 295 300
Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn
305 310 315 320
Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro
325 330 335
Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile
340 345 350
Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser
355 360 365
Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val
370 375 380
Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn
385 390 395 400
Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys
405 410 415
Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro
420 425 430
Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn
435 440 445
Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys
450 455 460
Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met
465 470 475 480
Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val
485 490 495
Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp
500 505 510
Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Pro Lys Glu
515 520 525
Ser Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro Glu Ser Leu Gly
530 535 540
Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg
545 550 555 560
Ile Thr Arg Thr Pro Glu Ile Thr Cys Val Val Leu Asp Leu Gly Arg
565 570 575
Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val
580 585 590
His Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr Tyr
595 600 605
Arg Val Val Ser Val Leu Pro Ile Glu His Gln Asp Trp Leu Thr Gly
610 615 620
Lys Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu Pro Ser Pro Ile
625 630 635 640
Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val
645 650 655
Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val
660 665 670
Thr Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp Val
675 680 685
Glu Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser Lys Tyr His Thr
690 695 700
Thr Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
705 710 715 720
Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr Cys
725 730 735
Ala Val Met His Glu Ala Leu Gln Asn His Tyr Thr Asp Leu Ser Leu
740 745 750
Ser His Ser Pro Gly Lys
755
<210> 18
<211> 590
<212> PRT
<213> Artificial Sequence
<220>
<223> Felis catus IL4R-IL13R (without signal sequence)
<400> 18
Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr
1 5 10 15
Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu
35 40 45
Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys
50 55 60
Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His
65 70 75 80
Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser
85 90 95
Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu
115 120 125
Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu
130 135 140
Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His
145 150 155 160
Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu
165 170 175
Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln
180 185 190
Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser
195 200 205
Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg
210 215 220
Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu
225 230 235 240
Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met
245 250 255
Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Gly Gly
260 265 270
Gly Ser Gly Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser
275 280 285
Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly
290 295 300
Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys
305 310 315 320
Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro
325 330 335
Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn
340 345 350
Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro
355 360 365
Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His
370 375 380
Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser
385 390 395 400
Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys
405 410 415
Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys
420 425 430
Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser
435 440 445
Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe
450 455 460
Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile
465 470 475 480
Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn
485 490 495
Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn
500 505 510
Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys
515 520 525
Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe
530 535 540
Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg
545 550 555 560
Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn
565 570 575
Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr
580 585 590
<210> 19
<211> 525
<212> PRT
<213> Artificial Sequence
<220>
<223> Equus caballus IL4R-IL13R (without signal sequence)
<400> 19
Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys
20 25 30
Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp
35 40 45
Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys
50 55 60
Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp
65 70 75 80
Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser
85 90 95
Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile
100 105 110
Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn
115 120 125
His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp
130 135 140
Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr
145 150 155 160
Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala
165 170 175
Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp
180 185 190
Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Gly Gly Gly
195 200 205
Ser Gly Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val
210 215 220
Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val
225 230 235 240
Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln
245 250 255
Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu
260 265 270
Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu
275 280 285
Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu
290 295 300
Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn
305 310 315 320
Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro
325 330 335
Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile
340 345 350
Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser
355 360 365
Phe Ala Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val
370 375 380
Gln Ile Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn
385 390 395 400
Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys
405 410 415
Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro
420 425 430
Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn
435 440 445
Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys
450 455 460
Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met
465 470 475 480
Val Pro Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val
485 490 495
Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp
500 505 510
Ser Gln Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr
515 520 525
<210> 20
<211> 780
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL13R-IL4R-IgGB (with signal sequence)
<400> 20
Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys
1 5 10 15
Val Leu Ser Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser
20 25 30
Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly
35 40 45
Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys
50 55 60
Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro
65 70 75 80
Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn
85 90 95
Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro
100 105 110
Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His
115 120 125
Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser
130 135 140
Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys
145 150 155 160
Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys
165 170 175
Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser
180 185 190
Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe
195 200 205
Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile
210 215 220
Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn
225 230 235 240
Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn
245 250 255
Asn Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys
260 265 270
Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe
275 280 285
Met Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg
290 295 300
Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn
305 310 315 320
Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly
325 330 335
Gly Ser Gly Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe
340 345 350
Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro
355 360 365
Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met
370 375 380
Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val
385 390 395 400
Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr
405 410 415
Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe
420 425 430
Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val
435 440 445
His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr
450 455 460
Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val
465 470 475 480
Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr
485 490 495
Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala
500 505 510
Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr
515 520 525
Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro
530 535 540
Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys
545 550 555 560
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
565 570 575
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
580 585 590
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
595 600 605
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
610 615 620
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
625 630 635 640
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
645 650 655
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
660 665 670
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
675 680 685
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
690 695 700
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
705 710 715 720
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
725 730 735
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
740 745 750
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
755 760 765
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
770 775 780
<210> 21
<211> 781
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R-IL13R-IgGB (with signal sequence)
<400> 21
Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys
1 5 10 15
Val Leu Ser Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe
20 25 30
Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro
35 40 45
Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met
50 55 60
Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val
65 70 75 80
Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr
85 90 95
Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe
100 105 110
Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val
115 120 125
His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr
130 135 140
Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val
145 150 155 160
Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr
165 170 175
Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala
180 185 190
Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr
195 200 205
Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro
210 215 220
Gly Gly Gly Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser
225 230 235 240
Val Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro
245 250 255
Glu Gly Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp
260 265 270
Asn Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu
275 280 285
Val Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser
290 295 300
Thr Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro
305 310 315 320
Pro Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val
325 330 335
Trp His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn
340 345 350
Thr Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu
355 360 365
Gly Lys Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile
370 375 380
Gly Cys Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln
385 390 395 400
His Ser Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro
405 410 415
Ser Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro
420 425 430
His Ile Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp
435 440 445
Lys Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu
450 455 460
Val Asn Asn Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu
465 470 475 480
Ala Lys Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile
485 490 495
Cys Phe Met Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg
500 505 510
Ile Arg Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp
515 520 525
Ser Asn Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr
530 535 540
Pro Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys
545 550 555 560
Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
565 570 575
Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr
580 585 590
Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser
595 600 605
Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg
610 615 620
Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile
625 630 635 640
Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn
645 650 655
Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
660 665 670
Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu
675 680 685
Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe
690 695 700
Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
705 710 715 720
Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly
725 730 735
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
740 745 750
Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn
755 760 765
His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
770 775 780
<210> 22
<211> 315
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL13R extracellular domain (without signal sequence)
<400> 22
Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro
20 25 30
Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile
165 170 175
Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu
195 200 205
Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr
305 310 315
<210> 23
<211> 207
<212> PRT
<213> Artificial Sequence
<220>
<223> Canis lupus IL4R extracellular domain (without signal sequence)
<400> 23
Ser Gly Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser
1 5 10 15
Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr
20 25 30
Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly
35 40 45
Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys
50 55 60
Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln
65 70 75 80
Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln
85 90 95
Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His
100 105 110
Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro
115 120 125
Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser
130 135 140
Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met
145 150 155 160
Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser
165 170 175
Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp
180 185 190
Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro
195 200 205
<210> 24
<211> 315
<212> PRT
<213> Artificial Sequence
<220>
<223> Felis catus IL13R extracellular domain (without signal sequence)
<400> 24
Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro
20 25 30
Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile
165 170 175
Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu
195 200 205
Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr
305 310 315
<210> 25
<211> 270
<212> PRT
<213> Artificial Sequence
<220>
<223> Felis catus IL4R extracellular domain (without signal sequence)
<400> 25
Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr
1 5 10 15
Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu
35 40 45
Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys
50 55 60
Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His
65 70 75 80
Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser
85 90 95
Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu
115 120 125
Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu
130 135 140
Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His
145 150 155 160
Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu
165 170 175
Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln
180 185 190
Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser
195 200 205
Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg
210 215 220
Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu
225 230 235 240
Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met
245 250 255
Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro
260 265 270
<210> 26
<211> 315
<212> PRT
<213> Artificial Sequence
<220>
<223> Equus caballus IL13R extracellular domain (without signal sequence)
<400> 26
Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro
20 25 30
Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile
165 170 175
Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu
195 200 205
Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr
305 310 315
<210> 27
<211> 205
<212> PRT
<213> Artificial Sequence
<220>
<223> Equus caballus IL4R extracellular domain (without signal sequence)
<400> 27
Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys
20 25 30
Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp
35 40 45
Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys
50 55 60
Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp
65 70 75 80
Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser
85 90 95
Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile
100 105 110
Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn
115 120 125
His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp
130 135 140
Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr
145 150 155 160
Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala
165 170 175
Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp
180 185 190
Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro
195 200 205
<210> 28
<211> 829
<212> PRT
<213> Artificial Sequence
<220>
<223> Feline IL13R-IL4R-IgG2 (without signal sequence)
<400> 28
Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro
20 25 30
Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile
165 170 175
Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu
195 200 205
Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly Gly Ser Gly
305 310 315 320
Ser Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp
325 330 335
Tyr Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn
340 345 350
Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser
355 360 365
Glu Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala
370 375 380
Cys Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu
385 390 395 400
His Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro
405 410 415
Ser Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro
420 425 430
Asn Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro
435 440 445
Glu Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser
450 455 460
Glu Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys
465 470 475 480
His Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg
485 490 495
Leu Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser
500 505 510
Gln Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr
515 520 525
Ser Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr
530 535 540
Arg Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val
545 550 555 560
Leu Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln
565 570 575
Met Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Ser
580 585 590
Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Cys Pro Lys
595 600 605
Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro
610 615 620
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
625 630 635 640
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
645 650 655
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
660 665 670
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
675 680 685
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
690 695 700
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
705 710 715 720
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
725 730 735
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
740 745 750
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
755 760 765
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
770 775 780
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
785 790 795 800
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
805 810 815
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
820 825
<210> 29
<211> 829
<212> PRT
<213> Artificial Sequence
<220>
<223> Feline IL4R-IL13R-IgG2 (without signal sequence)
<400> 29
Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr
1 5 10 15
Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys
20 25 30
Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu
35 40 45
Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys
50 55 60
Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His
65 70 75 80
Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser
85 90 95
Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn
100 105 110
Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu
115 120 125
Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu
130 135 140
Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His
145 150 155 160
Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu
165 170 175
Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln
180 185 190
Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser
195 200 205
Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg
210 215 220
Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu
225 230 235 240
Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met
245 250 255
Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Gly Gly
260 265 270
Gly Ser Gly Ser Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val
275 280 285
Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu
290 295 300
Gly Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn
305 310 315 320
Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val
325 330 335
Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr
340 345 350
Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro
355 360 365
Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp
370 375 380
His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr
385 390 395 400
Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly
405 410 415
Lys Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly
420 425 430
Cys Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His
435 440 445
Ser Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser
450 455 460
Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His
465 470 475 480
Ile Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys
485 490 495
Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val
500 505 510
Asn Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala
515 520 525
Lys Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys
530 535 540
Phe Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile
545 550 555 560
Arg Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser
565 570 575
Asn Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Ser
580 585 590
Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Cys Pro Lys
595 600 605
Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro
610 615 620
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
625 630 635 640
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
645 650 655
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
660 665 670
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
675 680 685
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
690 695 700
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
705 710 715 720
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
725 730 735
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
740 745 750
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
755 760 765
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
770 775 780
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
785 790 795 800
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
805 810 815
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
820 825
<210> 30
<211> 754
<212> PRT
<213> Artificial Sequence
<220>
<223> Equine IL13R-IL4R-IgG2 (without signal sequence)
<400> 30
Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn
1 5 10 15
Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro
20 25 30
Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys
35 40 45
Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu
50 55 60
Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp
65 70 75 80
Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp
85 90 95
Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser
100 105 110
Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr
115 120 125
Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln
130 135 140
Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala
145 150 155 160
Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile
165 170 175
Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val
180 185 190
Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu
195 200 205
Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn
210 215 220
Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln
225 230 235 240
Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn
245 250 255
Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro
260 265 270
Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr
275 280 285
Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln
290 295 300
Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr Gly Gly Gly Ser Gly
305 310 315 320
Ser Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp
325 330 335
Tyr Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn
340 345 350
Cys Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser
355 360 365
Asp Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val
370 375 380
Cys Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu
385 390 395 400
Asp Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro
405 410 415
Ser Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala
420 425 430
Ile Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys
435 440 445
Asn His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu
450 455 460
Asp Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro
465 470 475 480
Thr Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser
485 490 495
Ala Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu
500 505 510
Trp Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Asp Met
515 520 525
Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
530 535 540
Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg
545 550 555 560
Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro
565 570 575
Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala
580 585 590
Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val
595 600 605
Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe
610 615 620
Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala
625 630 635 640
Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu
645 650 655
Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys
660 665 670
Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser
675 680 685
Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln
690 695 700
Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu
705 710 715 720
Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His
725 730 735
Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu
740 745 750
Gly Lys
<210> 31
<211> 754
<212> PRT
<213> Artificial Sequence
<220>
<223> Equine IL4R-IL13R-IgG2 (without signal sequence)
<400> 31
Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr
1 5 10 15
Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys
20 25 30
Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp
35 40 45
Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys
50 55 60
Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp
65 70 75 80
Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser
85 90 95
Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile
100 105 110
Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn
115 120 125
His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp
130 135 140
Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr
145 150 155 160
Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala
165 170 175
Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp
180 185 190
Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Gly Gly Gly
195 200 205
Ser Gly Ser Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser
210 215 220
Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly
225 230 235 240
Val Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys
245 250 255
Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro
260 265 270
Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn
275 280 285
Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro
290 295 300
Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His
305 310 315 320
Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser
325 330 335
Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys
340 345 350
Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys
355 360 365
Ser Phe Ala Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser
370 375 380
Val Gln Ile Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe
385 390 395 400
Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile
405 410 415
Lys Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn
420 425 430
Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn
435 440 445
Asn Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys
450 455 460
Cys Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe
465 470 475 480
Met Val Pro Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg
485 490 495
Val Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn
500 505 510
Trp Ser Gln Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr Asp Met
515 520 525
Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
530 535 540
Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg
545 550 555 560
Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro
565 570 575
Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala
580 585 590
Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val
595 600 605
Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe
610 615 620
Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala
625 630 635 640
Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu
645 650 655
Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys
660 665 670
Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser
675 680 685
Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln
690 695 700
Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu
705 710 715 720
Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His
725 730 735
Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu
740 745 750
Gly Lys
<210> 32
<211> 305
<212> PRT
<213> Artificial Sequence
<220>
<223> Canine mini-IL13R ECD
<400> 32
Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr
1 5 10 15
Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr
20 25 30
Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala
35 40 45
Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys
50 55 60
Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser
65 70 75 80
Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser
85 90 95
Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys
100 105 110
Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr
115 120 125
Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp
130 135 140
Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn
145 150 155 160
Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys
165 170 175
Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr
180 185 190
Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln
195 200 205
Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser
210 215 220
Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr
225 230 235 240
Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe
245 250 255
Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu
260 265 270
Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu
275 280 285
Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser
290 295 300
Ile
305
<210> 33
<211> 184
<212> PRT
<213> Artificial Sequence
<220>
<223> Canine mini-IL4R ECD
<400> 33
Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr Ile Ser Thr Ser
1 5 10 15
Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys Ser Ala Glu Leu
20 25 30
Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu Asn His Thr Cys
35 40 45
Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys Ser Met Pro Ile
50 55 60
Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp Leu Trp Ala Gly
65 70 75 80
Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser Lys His Val Lys
85 90 95
Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn Ile Ser His Thr
100 105 110
Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu Asn His Leu His
115 120 125
Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp Asn Asp Pro Glu
130 135 140
Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro Thr Leu Arg Leu
145 150 155 160
Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser Ala Arg Val Arg
165 170 175
Ala Trp Ala Gln Thr Tyr Asn Ser
180
<210> 34
<211> 305
<212> PRT
<213> Artificial Sequence
<220>
<223> Feline mini-IL13R ECD
<400> 34
Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr
1 5 10 15
Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr
20 25 30
Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala
35 40 45
Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys
50 55 60
Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser
65 70 75 80
Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser
85 90 95
Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys
100 105 110
Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr
115 120 125
Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asn
130 135 140
Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn
145 150 155 160
Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys
165 170 175
Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr
180 185 190
Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln
195 200 205
Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser
210 215 220
Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr
225 230 235 240
His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe
245 250 255
Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Ile Leu
260 265 270
Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu
275 280 285
Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln Ala Met Ser
290 295 300
Ile
305
<210> 35
<211> 266
<212> PRT
<213> Artificial Sequence
<220>
<223> Feline mini-IL4R ECD
<400> 35
Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr Phe Ser Thr Ser
1 5 10 15
Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys Ser Ala Glu Leu
20 25 30
Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu Asn Arg Thr Cys
35 40 45
Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys Ser Met Leu Met
50 55 60
Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His Leu Trp Ala Gly
65 70 75 80
Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser Ser His Val Lys
85 90 95
Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn Val Ser His Thr
100 105 110
Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu Asn His Leu His
115 120 125
Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu Asp Asp Pro Thr
130 135 140
Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His Leu Leu Gly Leu
145 150 155 160
Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu Pro Pro Trp Leu
165 170 175
Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln Cys Ala Val Ile
180 185 190
Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser Arg Gly Gly Arg
195 200 205
Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg Tyr Val Ser Val
210 215 220
Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu Cys Pro Gly Thr
225 230 235 240
Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met Ser Pro Asp Pro
245 250 255
Ser Ala Phe His Ser Ile Asp Tyr Glu Pro
260 265
<210> 36
<211> 305
<212> PRT
<213> Artificial Sequence
<220>
<223> Equine mini-IL13R ECD
<400> 36
Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr
1 5 10 15
Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro Asn Cys Ser
20 25 30
Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys Lys Ile Ala
35 40 45
Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys
50 55 60
Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser
65 70 75 80
Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp Pro Glu Ser
85 90 95
Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys
100 105 110
Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr Asn Tyr Thr
115 120 125
Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp
130 135 140
Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Glu
145 150 155 160
Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile Met Val Lys
165 170 175
Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val Pro Leu Thr
180 185 190
Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu Phe Phe Gln
195 200 205
Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser
210 215 220
Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr
225 230 235 240
Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn Pro Glu Phe
245 250 255
Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu
260 265 270
Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr Asn Lys Leu
275 280 285
Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser
290 295 300
Ile
305
<210> 37
<211> 201
<212> PRT
<213> Artificial Sequence
<220>
<223> Equine mini-IL4R ECD
<400> 37
Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr Ile Ser Ala Ser
1 5 10 15
Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys Ser Ala Gln Leu
20 25 30
Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp Asn Leu Thr Cys
35 40 45
Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys Arg Met Leu Met
50 55 60
Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp Leu Trp Ala Gly
65 70 75 80
Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser Arg His Val Lys
85 90 95
Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile Ser His Thr Trp
100 105 110
Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn His Leu Trp Ser
115 120 125
Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp Asp Pro Thr Asp
130 135 140
Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr Leu Arg Val Thr
145 150 155 160
Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala Arg Val Lys Ala
165 170 175
Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp Ser Pro Ser Thr
180 185 190
Thr Trp His Asn Tyr Tyr Glu Gln Pro
195 200
<210> 38
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 38
Gly Ser Gly Ser
1
<210> 39
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 39
Gly Ser Gly Ser Gly Ser
1 5
<210> 40
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 40
Gly Gly Ser Gly Gly Ser
1 5
<210> 41
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 41
Gly Gly Ser Gly Gly Ser Gly Gly Ser
1 5
<210> 42
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 42
Gly Gly Gly Ser
1
<210> 43
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 43
Gly Gly Gly Ser Gly Gly Gly Ser
1 5
<210> 44
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 44
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
1 5 10
<210> 45
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 45
Gly Ser Ser Gly Ser Ser
1 5
<210> 46
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 46
Gly Ser Ser Gly Ser Ser Gly Ser Ser
1 5
<210> 47
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 47
Gly Gly Ser Ser
1
<210> 48
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 48
Gly Gly Ser Ser Gly Gly Ser Ser
1 5
<210> 49
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> GS linker
<400> 49
Gly Gly Ser Ser Gly Gly Ser Ser Gly Gly Ser Ser
1 5 10
Claims (41)
- IL13R 폴리펩타이드의 세포외 도메인 및 IL4R 폴리펩타이드의 세포외 도메인(extracellular domain)을 포함하는 연속된(contiguous) 폴리펩타이드로서, 여기서 IL13R 및 IL4R 폴리펩타이드는 반려 동물 종으로부터 유래되는, 연속된 폴리펩타이드.
- 제1항에 있어서, 식 (I) IL13R-L1-IL4R-L2-FP 또는 식 (II) IL4R-L1-IL13R-L2-FP를 포함하는 연속된 폴리펩타이드로서, 여기서:
a. IL13R은 반려 동물 종으로부터 유래된 IL13R 폴리펩타이드의 세포외 도메인이고,
b. IL4R는 반려 동물 종으로부터 유래된 IL4R 폴리펩타이드의 세포외 도메인이며,
c. L1은 제1의 임의의 링커이고,
d. L2는 제2의 임의의 링커이며,
e. FP는 융합 파트너인, 연속된 폴리펩타이드. - 제1항 또는 제2항에 있어서, 연속된 폴리펩타이드가 생물층 간섭법(biolayer interferometry)에 의해 측정된 바와 같이, 반려 동물 종의 IL13에 5 x 10-6 M 미만, 1 x 10-6 M 미만, 5 x 10-7 M 미만, 1 x 10-7 M 미만, 5 x 10-8 M 미만, 1 x 10-8 M 미만, 5 x 10-9 M 미만, 1 x 10-9 M 미만, 5 x 10-10 M 미만, 1 x 10-10 M 미만, 5 x 10-11 M 미만, 1 x 10-11 M 미만, 5 x 10-12 M 미만, 또는 1 x 10-12 M 미만의 해리 상수(Kd)로 결합하는, 연속된 폴리펩타이드.
- 제1항 내지 제3항 중 어느 한 항에 있어서, 연속된 폴리펩타이드가 생물층 간섭법에 의해 측정된 바와 같이, 반려 동물 종의 IL4에 5 x 10-6 M 미만, 1 x 10-6 M 미만, 5 x 10-7 M 미만, 1 x 10-7 M 미만, 5 x 10-8 M 미만, 1 x 10-8 M 미만, 5 x 10-9 M 미만, 1 x 10-9 M 미만, 5 x 10-10 M 미만, 1 x 10-10 M 미만, 5 x 10-11 M 미만, 1 x 10-11 M 미만, 5 x 10-12 M 미만, 또는 1 x 10-12 M 미만의 해리 상수로 결합하는, 연속된 폴리펩타이드.
- 제1항 내지 제4항 중 어느 한 항에 있어서, 연속된 폴리펩타이드가 반려 동물 종에서 IL13 및/또는 IL4 신호전달을 감소시키는, 연속된 폴리펩타이드.
- 제1항 내지 제5항 중 어느 한 항에 있어서, 반려 동물 종이 개과(canine), 고양이과(feline), 또는 말과(equine)인, 연속된 폴리펩타이드.
- 제1항 내지 제6항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 85% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제7항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 90% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제8항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 95% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제9항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22, 서열 번호: 24, 서열 번호: 26, 서열 번호: 32, 서열 번호: 34, 또는 서열 번호: 36의 아미노산 서열에 대해 적어도 98% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제10항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22의 18번 위치에 상응하거나, 서열 번호: 24의 18번 위치에 상응하거나, 서열 번호: 26의 18번 위치에 상응하는 위치에서 시스테인을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제11항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22의 18번 위치에서, 서열 번호: 24의 18번 위치에서, 서열 번호: 26의 18번 위치에서, 서열 번호: 32의 15번 위치에서, 서열 번호: 34의 15번 위치에서, 또는 서열 번호: 36의 15번 위치에서 시스테인을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제12항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 32, 서열 번호: 34, 및 서열 번호: 36으로부터 선택된 아미노산 서열을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제13항 중 어느 한 항에 있어서, IL13R 폴리펩타이드의 세포외 도메인이 서열 번호: 22, 서열 번호: 24, 및 서열 번호: 26으로부터 선택된 아미노산 서열을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제14항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 85% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제15항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 90% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제16항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산에 대해 적어도 95% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제17항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 23, 서열 번호: 25, 서열 번호: 27, 서열 번호: 33, 서열 번호: 35, 또는 서열 번호: 37의 아미노산 서열에 대해 적어도 98% 동일한, 연속된 폴리펩타이드.
- 제1항 내지 제18항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 33, 서열 번호: 35, 및 서열 번호: 37로부터 선택된 아미노산 서열을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제19항 중 어느 한 항에 있어서, IL4R 폴리펩타이드의 세포외 도메인이 서열 번호: 23, 서열 번호: 25, 및 서열 번호: 27로부터 선택된 아미노산 서열을 포함하는, 연속된 폴리펩타이드.
- 제2항 내지 제22항 중 어느 한 항에 있어서, L1 및 L2는, 존재하는 경우, 각각 독립적으로 G, GG, GGG, S, SS, SSS, GS, GSGS(서열 번호: 38), GSGSGS(서열 번호: 39), GGS, GGSGGS(서열 번호: 40), GGSGGSGGS(서열 번호: 41), GGGS(서열 번호: 42), GGGSGGGS(서열 번호: 43), GGGSGGGSGGGS(서열 번호: 44), GSS, GSSGSS(서열 번호: 45), GSSGSSGSS(서열 번호: 46), GGSS(서열 번호: 47), GGSSGGSS(서열 번호: 48), 및 GGSSGGSSGGSS(서열 번호: 49)로부터 선택된 아미노산 서열을 포함하는, 연속된 폴리펩타이드.
- 제2항 내지 제21항 중 어느 한 항에 있어서, 융합 파트너가 Fc, 알부민, 및 알부민 결합 단편을 포함하는, 연속된 폴리펩타이드.
- 제22항에 있어서, Fc가 (a) IgG-A, IgG-B, IgG-C, 및 IgG-D 불변 영역으로부터 선택된 개과 중쇄 불변 영역; (b) IgG1, IgG2a, 및 IgG2b 불변 영역으로부터 선택된 고양이과 중쇄 불변 영역; 또는 (c) IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 및 IgG7 불변 영역으로부터 선택된 말과 중쇄 불변 영역인, 연속된 폴리펩타이드.
- 제1항 내지 제23항 중 어느 한 항에 있어서, 연속된 폴리펩타이드가 서열 번호: 13, 서열 번호: 14, 서열 번호: 15, 서열 번호: 16, 서열 번호: 17, 서열 번호 18, 서열 번호: 19, 서열 번호: 20, 서열 번호: 21, 서열 번호: 28, 서열 번호: 29, 서열 번호: 30, 및 서열 번호: 31로부터 선택된 서열을 포함하는, 연속된 폴리펩타이드.
- 제1항 내지 제24항 중 어느 한 항에 따른 연속된 폴리펩타이드를 암호화하는, 단리된 핵산.
- 제25항에 따른 핵산을 포함하는 숙주 세포.
- 제26항에 따른 숙주 세포를 배양하는 단계 및 연속된 폴리펩타이드를 단리하는 단계를 포함하는, 연속된 폴리펩타이드의 생산 방법.
- 제1항 내지 제24항 중 어느 한 항에 따른 연속된 폴리펩타이드 및 약제학적으로 허용되는 담체를 포함하는 약제학적 조성물.
- 반려 동물 종에게 치료학적 유효량의 제1항 내지 제24항 중 어느 한 항에 따른 연속된 폴리펩타이드 또는 제28항에 따른 약제학적 조성물을 투여하는 단계를 포함하는, IL13 및/또는 IL4-유도된 상태를 갖는 반려 동물 종을 치료하는 방법.
- 제29항에 있어서, 반려 동물 종이 개과, 고양이과, 또는 말과인 방법.
- 제29항 또는 제30항에 있어서, IL13 및/또는 IL4-유도된 상태가 소양증 또는 알레르기 상태, 예를 들면, 아토피성 피부염, 소양증, 천식, 건선, 피부경화증, 또는 습진인 방법.
- 제29항 내지 제32항 중 어느 한 항에 있어서, 연속된 폴리펩타이드 또는 약제학적 조성물이 비경구적으로 투여되는 방법.
- 제29항 내지 제32항 중 어느 한 항에 있어서, 연속된 폴리펩타이드 또는 약제학적 조성물이 근육내 경로, 복강내 경로, 뇌척수내 경로, 피하 경로, 동맥내 경로, 활액내 경로, 수막내 경로, 또는 흡입 경로에 의해 투여되는 방법.
- 제29항 내지 제33항 중 어느 한 항에 있어서, 방법이 Jak 억제제, PI3K 억제제, AKT 억제제, 또는 MAPK 억제제를 투여함을 추가로 포함하는 방법.
- 제29항 내지 제34항 중 어느 한 항에 있어서, 방법이 항-IL17 항체, 항-IL31 항체, 항-TNFα 항체, 항-CD20 항체, 항-CD19 항체, 항-CD25 항체, 항-IL4 항체, 항-IL13 항체, 항-IL23 항체, 항-IgE 항체, 항-CD11α 항체, 항-IL6R 항체, 항-α4-인테그린 항체, 항-IL12 항체, 항-IL1β 항체, 및 항-BlyS 항체로부터 선택된 하나 이상의 항체를 투여함을 추가로 포함하는 방법.
- 세포를 IL13 및/또는 IL4에 대한 연속된 폴리펩타이드의 결합을 허용하는 조건하에서 제1항 내지 제24항 중 어느 한 항에 따른 연속된 폴리펩타이드 또는 제28항에 따른 약제학적 조성물에 노출시킴으로써, (a) 천연의 IL13 수용체 및/또는 천연의 IL-4 수용체에 대한 IL/4 및/또는 IL-13의 결합을 감소시키고 IL13- 및/또는 IL-4-매개된 신호전달을 감소시킴을 포함하는, 세포내에서 IL13 및/또는 IL4 신호전달 활성을 감소시키는 방법.
- 제36항에 있어서, 세포가 연속된 폴리펩타이드 또는 약제학적 조성물에 생체외(ex vivo)에서 노출되는 방법.
- 제36항에 있어서, 세포가 연속된 폴리펩타이드 또는 약제학적 조성물에 생체내에서 노출되는 방법.
- 제36항 내지 제38항 중 어느 한 항에 있어서, 세포가 개과 세포, 고양이과 세포, 또는 말과 세포인 방법.
- 샘플을 IL13 및/또는 IL4에 대한 연속된 폴리펩타이드의 결합을 허용하는 조건 하에서 제1항 내지 제24항 중 어느 한 항에 따른 연속된 폴리펩타이드 또는 제28항에 따른 약제학적 조성물과 접촉시키고, 복합체가 샘플 속의 연속된 폴리펩타이드와 IL13 및/또는 IL4 사이에 형성되는지를 검출함을 포함하여, 반려 동물 종으로부터의 샘플 속에서 IL13 또는 IL4를 검출하는 방법.
- 제40항에 있어서, 샘플이 개과, 고양이과, 또는 말과로부터 수득된 생물학적 샘플인 방법.
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| PCT/US2018/028507 WO2018195388A1 (en) | 2017-04-21 | 2018-04-20 | Il4/il13 receptor molecule for veterinary use |
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| EP3668536A4 (en) * | 2017-08-15 | 2021-05-26 | Kindred Biosciences, Inc. | VARIANTS OF IGG FC FOR VETERINARY USE |
| EP3870601A4 (en) * | 2018-10-25 | 2022-07-13 | Kindred Biosciences, Inc. | IL4/IL13 RECEPTOR MOLECULE FOR USE IN VETERINARY MEDICINE |
| EP4139344A1 (en) * | 2020-04-22 | 2023-03-01 | Kindred Biosciences, Inc. | Il4/il13 receptor molecules for veterinary use |
| WO2021249555A1 (en) * | 2020-06-12 | 2021-12-16 | Beijing Vdjbio Co., Ltd. | Fusion polypeptide |
| WO2023021170A2 (en) | 2021-08-20 | 2023-02-23 | Intervet International B.V. | Fusion proteins for treating atopic dermatitis |
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| US6743604B1 (en) * | 1996-12-13 | 2004-06-01 | Smithkline Beecham Corporation | Substances and their uses |
| US6472179B2 (en) * | 1998-09-25 | 2002-10-29 | Regeneron Pharmaceuticals, Inc. | Receptor based antagonists and methods of making and using |
| US7083949B2 (en) * | 1998-09-25 | 2006-08-01 | Regeneron Pharmaceuticals, Inc. | Receptor based antagonists and methods of making and using |
| BR9916209A (pt) * | 1998-12-14 | 2001-12-26 | Genetics Inst | Cadeia de receptor de citocina |
| EP1268794A2 (en) * | 2000-04-07 | 2003-01-02 | Heska Corporation | Compositions and methods related to canine igg and canine il-13 receptors |
| CA2550651C (en) * | 2003-12-23 | 2018-01-23 | Tanox, Inc. | Novel anti-il 13 antibodies and uses thereof |
| JP2008504806A (ja) * | 2004-02-27 | 2008-02-21 | リジェネロン・ファーマシューティカルズ・インコーポレイテッド | Ｉｌ−４／ｉｌ−１３特異的ポリペプチドおよびその治療上の使用 |
| CN1922204A (zh) * | 2004-02-27 | 2007-02-28 | 瑞泽恩制药公司 | Il－4/il－13特异性的多肽和其治疗应用 |
| EP2674440B1 (en) * | 2005-12-16 | 2019-07-03 | IBC Pharmaceuticals, Inc. | Multivalent immunoglobulin-based bioactive assemblies |
| MX2009005725A (es) * | 2006-12-11 | 2009-08-24 | Wyeth Corp | Metodos y composiciones para tratamiento y monitoreo de tratamiento de trastornos asociados con il-13. |
| WO2009061911A1 (en) * | 2007-11-06 | 2009-05-14 | Wyeth | Assays and methods for evaluating multimeric complexes |
| EP2702069A4 (en) * | 2011-04-29 | 2015-04-29 | Janssen Biotech Inc | IL4 / IL13 BINDING REPEAT PROTEINS AND USES THEREOF |
| JP6163162B2 (ja) * | 2011-12-13 | 2017-07-12 | ピエリス ファーマシューティカルズ ゲーエムベーハー | Ｉｌ−４および／またはｉｌ−１３のそれぞれの受容体への結合を阻害することによる特定の障害の予防または治療のための方法 |
| EP2850093B1 (en) * | 2012-01-27 | 2019-01-09 | The Board of Trustees of the Leland Stanford Junior University | Therapeutic il-13 polypeptides |
| LT2888281T (lt) * | 2012-08-21 | 2018-12-27 | Sanofi Biotechnology | Astmos gydymo arba profilaktikos būdai, skiriant il-4r antagonistą |
| JP6938383B2 (ja) * | 2015-04-02 | 2021-09-22 | インターベット インターナショナル ベー． フェー． | イヌインターロイキン４受容体アルファに対する抗体 |
| KR20180034619A (ko) * | 2015-07-29 | 2018-04-04 | 나노틱스 엘엘씨 | 가용성 생체분자를 소거하기 위한 모듈 조성물 및 이와 관련된 방법 |
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| RU2019137211A3 (ko) | 2021-09-09 |
| AU2018254542A1 (en) | 2019-11-21 |
| US20200048325A1 (en) | 2020-02-13 |
| RU2019137211A (ru) | 2021-05-21 |
| EP3612555A4 (en) | 2020-12-30 |
| JP2023078158A (ja) | 2023-06-06 |
| AU2018254542B2 (en) | 2022-12-22 |
| CA3059731A1 (en) | 2018-10-25 |
| BR112019021812A2 (pt) | 2020-05-26 |
| CN110770250A (zh) | 2020-02-07 |
| MX2019012570A (es) | 2019-12-02 |
| AU2023201726A1 (en) | 2023-04-27 |
| WO2018195388A1 (en) | 2018-10-25 |
| EP3612555A1 (en) | 2020-02-26 |
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