A method of improving efficacy of biological response-modifying proteins and the example muteins

생리활성 조절 단백질의 효능 향상 방법 및 그 예시변이체들

Abstract

본 발명은 수용체, 리간드 또는 기질과 결합하여 생리활성 조절작용을 나타내는 단백질의 결합 도메인내 아미노산 중 페닐알라닌을 발린으로 치환한 단백질 변이체, 이 변이체를 코딩하는 DNA, 이 DNA가 벡터에 작동가능하게 연결된 재조합 발현 벡터, 이 재조합 발현 벡터로 형질전환 또는 형질감염된 숙주세포, 이 숙주세포를 배양하고 이의 배양물로부터 상기 단백질 변이체를 분리하는 단계를 포함한 단백질 변이체의 제조방법에 관한 것이다. 또한, 본 발명은 상기 단백질 변이체 및 약제학적으로 허용되는 담체를 포함하는 약제학적 조성물에 관한 것이다.
PURPOSE: A method for improving efficacy of biological response-modifying proteins and the muteins of the same proteins are provided, which muteins of the biological response-modifying proteins have improved binding affinity and biological-activation inducing activity, and do not result in an autoimmunity response. CONSTITUTION: The muteins of the biological response-modifying proteins which are bound with a receptor, ligand or substrate to modify their biological response are prepared by substituting an amino acid phenylalanine in a binding domain with an amino acid valine, wherein the proteins are cytokines comprising CNTF, EPO, FIt3L, G-CSF, GM-CSF, GRH, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12p35, LPT, LIF, M-CSF, OSM, PL, SCF and TPO; and the cytokine is interferon comprising IFN-α2A, IFN-α2B, IFN-β, IFN-γ, IFN-¥ø and IFN-τ. Genes encoding the muteins of the biological response-modifying proteins are provided. A method for producing the muteins of the biological response-modifying proteins comprises the steps of: constructing an expression vector containing the gene encoding a mutein of the biological response-modifying proteins; transforming a host cell with the recombinant expression vector; and culturing the transformed host cell in a medium and isolating the mutein.

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