KR920701255A - 에리쓰로포이에틴 동형체(eryihropoietin isoform) - Google Patents

에리쓰로포이에틴 동형체(eryihropoietin isoform)

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KR920701255A
KR920701255A KR1019910700600A KR910700600A KR920701255A KR 920701255 A KR920701255 A KR 920701255A KR 1019910700600 A KR1019910700600 A KR 1019910700600A KR 910700600 A KR910700600 A KR 910700600A KR 920701255 A KR920701255 A KR 920701255A
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erythropoietin
isoform
per molecule
sialic acids
mixture
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토마스 웨인 스트릭크랜드
토마스 에드워드 바이른
스티븐 조지 엘리오트
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스티븐 엠. 오드레
암젠 인코포레이티드
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    • C—CHEMISTRY; METALLURGY
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    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/505—Erythropoietin [EPO]
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

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Abstract

내용 없음

Description

에리쓰로포이에틴 동형체(ERYIHROPOIETIN ISOFORM)
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 재조합 에리쓰로포이에틴 동형체들의 분리의 분석적 등전집중겔을 나타내고 있다. 겔의 레인(lane)(1-11)들은 레인(1)의 약산성(높은 pI)으로 부터 레인(11)의 보다 강한 산성(낮은 pI)까지에 걸쳐있는 동형체들을 나타낸다. 동형체(9-14)들읜 혼합물을 함유하는 정제된 재조합 에리쓰로포이에틴의 겔의 최좌측(far left)레인과 최우측레인들에 나타나있다. 제23도는 에리쓰로포이에틴 동형체 당 시알산의 수와 에리쓰로포이에틴 폴리펩티드의 ㎎당 단위로서 표시된 각 동형체의 생체내에서의 비활성 사이의 관계를 나타내고 있따. 제2A도에서, 각 에리쓰로포이에틴 동형체의 농도는 브래드포드 단백질효력검정(Bradford protein assay)에 의해 결정된 것이고, 제2B도에서, 에리쓰로포이에틴 동형체의 농도는 280nm에서의 흡수율에 의해 결정된 것이며, 제2C도에서 에리쓰로포이에틴 동형체의 농도는 방사성면역측정법(Radiommunoassay:RIA)에 의해 결정된 것이다. 제3도는 다른 조건하에서 음이온교환 크로마토그래피에 의해 제조된 재조합 에리쓰로포이에틴 동형체들의 혼합물들을 구분하는 분석적 등전집중겔을 나타낸다. 겔 레인(1-6)들은 큐-세파로스 패스트 플로우 컬럼(Q-Separose fast flow column)을 각각 ph4.7의 150mM 초산, 완충시키지 않은 150mM 초산, pH4.7의 200mM 초산, pH4.7의 250/mM 초산, pH4.7의 300mM 초산 또는 완충시키지 않은 300mM 초산으로 세척한 후 고농도열 수용액(high salt)에서 용리시킨 에리쓰로포이에틴 동형체들을 나타낸다. 디에틸아미노에틸-아가로스(DEAE-Agarose)크로마토그래프를 큐-세파로스크로마토그래피로 대체한 것을 제외하고는 위에서의 레이와 그의 동료들의 특허의 실시예2에서 설명된 절차를 사용하여 얻어진 바와 같은 동형체들의 혼합물을 포함하는 정제된 재조합 에리쓰로포이에틴이 또한 겔의 최좌측 레인에 나타나 있다.

Claims (34)

  1. 에리쓰로포이에틴 동형체.
  2. 제1항에 있어서, 상기 동형체는 비인간의 진핵숙주세포내의 외인성 DNA 시퀀스의 형질발현의 생성물인 에리쓰로포이에틴 동형체.
  3. 제1항에 있어서, 상기 동형체는 인간의 에리쓰로포이에틴의 1-165 또는 1-666의 아미노산 시퀀스를 갖는 에리쓰로포이에틴을 포함하는 에리쓰로포이에틴 동형체.
  4. 제1항에 있어서, 에리쓰로포이에틴 분자당 1-14까지의 그룹에서 선택된 특정 수의 시알산을 가지는 에리쓰로포이에틴 동형체.
  5. 제1항에 있어서, 에리쓰로포이에틴 분자당 14개 이상의 시알산을 가지는 에리쓰로포이에틴 동형체.
  6. 제1항에 있어서, 에리쓰로포이에틴 분자당 14개의 시알산을 가지는 에리쓰로포이에틴 동형체.
  7. 제1항에 있어서, 에리쓰로포이에틴 분자당 13개의 시알산을 가지는 에리쓰로포이에틴 동형체.
  8. 제1항에 있어서, 에리쓰로포이에틴 분자당 10개의 시알산을 가지는 에리쓰로포이에틴 동형체.
  9. 제2항에 있어서, 상기 진핵숙주세포가 CHO(Chinese Hamster Ovary) 세포인 에리쓰로포이에틴 동형체.
  10. 제약학적으로 유효한 양의 상기 제1항의 에리쓰로포이에틴 동형체와 제약학적으로 수용가능한 희석제, 보조제 및 담체를 포함하는 제약학적 조성물.
  11. 4가지 이하의 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  12. 에리쓰로포이에틴 분자당 12개 이하의 시알산을 갖는 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  13. 제12항에 있어서, 에리쓰로포이에틴 분자당 9,10 및 11개의 시알산을 갖는 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  14. 에리쓰로포이에틴 분자당 11개 이상의 시알산을 갖는 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  15. 제14항에 있어서, 에리쓰로포이에틴 분자당 13-14개의 시알산을 갖는 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  16. 에리쓰로포이에틴 분자당 특정수의 시알산을 갖는 에리쓰로포이에틴 분자들로 실질적으로 구성되는 에리쓰로포이에틴.
  17. 에리쓰로포이에틴 분자당 1-14로 구성된 그룹에서 선택된 특정수의 시알산을 갖는 에리쓰로포이에틴 분자들로 실질적으로 구성되는 에리쓰로포이에틴.
  18. 에리쓰로포이에틴 분자당 14개 이상의 시알산을 갖는 에리쓰로포이에틴 분자들을 포함하는 에리쓰로포이에틴.
  19. 제16항에 있어서, 에리쓰로포이에틴 분자당 14개의 시알산을 갖는 에리쓰로포이에틴.
  20. 제16항에 있어서, 에리쓰로포이에틴 분자당 13개의 시알산을 갖는 에리쓰로포이에틴.
  21. 제16항에 있어서, 에리쓰로포이에틴 분자당 10개의 시알산을 갖는 에리쓰로포이에틴.
  22. 제16항에 있어서, 상기 분자들은 비인간의 진핵숙주세포 내에서 외인성 DNA 시퀀스의 형질발현의 생성물인 에리쓰로포이에틴.
  23. 제15항에 있어서, 상기 에리쓰로포이에틴은 인간의 에리쓰로포이에틴의 아미노산 시퀀스를 가지는 에리쓰로포이에틴.
  24. 제약학적으로 유효한 양의 상기 제15항의 에리쓰로포이에틴 및 제약학적으로 수용가능한 희석제, 보조제 및 담체를 포함하는 제약학적 조성물.
  25. 에리쓰로포이에틴 분자당 4 이하의 특정수의 시알산을 갖는 에리쓰로포이에틴 분자들의 혼합물을 포함하는 조성물.
  26. 에리쓰로포이에틴 분자당 12개 이하의 시알산을 갖는 에리쓰로포이에틴 분자들의 혼합물을 포함하는 조성물.
  27. 에리쓰로포이에틴 분자당 11개 이상의 시알산을 갖는 에리쓰로포이에틴 분자들의 혼합물을 포함하는 조성물.
  28. 제27항에 있어서, 에리쓰로포이에틴 분자당 13-14개의 시알산을 갖는 에리쓰로포이에틴 동형체들의 혼합물을 포함하는 조성물.
  29. (가) 정제된 에리쓰로포이에틴을 분리용등전집중법에 적용시키고, (나) 겔로부터 단일의 동형체를 용리시키는 단계들을 포함하는, 에리쓰로포이에틴 동형체의 제조방법.
  30. 에리쓰로포이에틴을 함유하는 물질을 이온교환크로마토그래피에 적용시켜 분자당 전결정된 수 이상의 시알산을 갖는 에리쓰로포이에틴 분자들의 혼합물의 제조방법.
  31. 에리쓰로포이에틴을 함유하는 물질을 크로마토포커싱에 적용시켜 분자당 전결정된 수 이상의 시알산을 갖는 에리쓰로포이에틴 분자들의 혼합물의 제조방법.
  32. 치료학적으로 유효한 양의 제26항에 따른 조성물을 투약함을 포함하는 포유동물의 적혈구용적비를 증가시키는 방법.
  33. 〔Asn69〕EPO, 〔Asn125, Ser127〕EPO, 〔Thr125〕EPO, 및 〔Pro|124, Thr125〕EPO로 구성된 그룹으로부터 선택된 인간의 에리쓰로포이에틴의 유사체.
  34. 〔Asn69〕EPO, 〔Asn125, Ser127〕EPO, 〔Thr125〕EPO, 및 〔Pro|124, Thr125〕EPO로 구성된 그룹으로부터 선택된 인간의 에리쓰로포이에틴의 유사체를 암호화하는 DNA 시퀀스로 실질적으로 구성된 정제되고 단리된 시퀀스.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019910700600A 1989-10-13 1990-10-09 에리트로포이에틴 유사체와 그를 포함하는 제약학적 조성물 KR100221066B1 (ko)

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US42144489A 1989-10-13 1989-10-13
US421444 1989-10-13
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PCT/US1990/005758 WO1991005867A1 (en) 1989-10-13 1990-10-09 Erythropoietin isoforms

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KR1019910700600A KR100221066B1 (ko) 1989-10-13 1990-10-09 에리트로포이에틴 유사체와 그를 포함하는 제약학적 조성물

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ZA908166B (en) 1991-08-28
LV12575B (en) 2001-03-20
HK1010398A1 (en) 1999-06-17
DE69033301T2 (de) 2000-04-20
KR100221066B1 (ko) 1999-10-01
CA2165694A1 (en) 1991-04-14
EP0428267A3 (en) 1991-08-14
CZ497290A3 (cs) 2000-08-16
DK0668351T3 (da) 1999-12-20
EP0428267B2 (en) 2004-12-08
IL95971A0 (en) 1991-07-18
CZ291471B6 (cs) 2003-03-12
WO1991005867A1 (en) 1991-05-02
DE69029370T2 (de) 1997-06-05
NO301832B1 (no) 1997-12-15
IL125175A0 (en) 1999-03-12
DK0428267T3 (da) 1997-06-09
EP0428267B1 (en) 1996-12-11
PT95586A (pt) 1991-09-30
EP0428267A2 (en) 1991-05-22
DE69029370T3 (de) 2006-02-23
CN1063796C (zh) 2001-03-28
CA2027635A1 (en) 1991-04-14
GR3032089T3 (en) 2000-03-31
ATE184914T1 (de) 1999-10-15
JP2983629B2 (ja) 1999-11-29
LV12575A (en) 2000-11-20
DE69029370C5 (de) 2010-03-25
ES2097753T3 (es) 1997-04-16
NO912281L (no) 1991-06-13
CZ291488B6 (cs) 2003-03-12
DK0428267T4 (da) 2005-03-14

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