JP7446474B2 - 小分子標的を同定するためのハイスループットスクリーニング方法 - Google Patents
小分子標的を同定するためのハイスループットスクリーニング方法 Download PDFInfo
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Description
[0001] 本出願は、2020年5月11日に出願された米国仮特許出願第63/023,181号に対する優先権を主張し、その全体が参照により本明細書に組み込まれる。
[0002] 細胞内の生物学的プロセスを標的として薬理学的な介入を行うことは、創薬の中心的な目標である。特定の標的タンパク質に対する阻害薬を同定するプロセスは、標的に対する高い親和性、標的効果に対する高い効力及び選択性、及び毒性及び意図しないオフターゲット効果を最小限に抑えながら、目的の組織で十分に高い薬物濃度を維持して所望の薬理学的効果を維持する用量を特定することの需要を満たさなければならない。小分子は、原形質膜を通過し、広範囲の組織及び作用部位にアクセスし、複数の標的に同時に影響を及ぼし、経済的に大規模で産生することができる能力を有するため、細胞内標的の調節の魅力的な候補である。
[0011] いくつかの実施形態では、タンパク質間相互作用をアッセイするための方法が提供され、この方法は、第1の複数の組換え一倍体酵母細胞の表面上に発現及び提示された複数のポリペプチドユビキチンリガーゼ種を提供することであって、第1の複数のポリペプチドユビキチンリガーゼ種が、野生型ポリペプチドユビキチンリガーゼ種と、変異誘発によって1つ又は複数のアミノ酸残基位置で改変された変異ポリペプチドユビキチンリガーゼ種とのライブラリーを含むこと;第2の複数の組換え一倍体酵母細胞の表面上に発現及び提示された複数のポリペプチド基質種を提供することであって、複数のポリペプチド基質種が、野生型ポリペプチド基質種と、変異誘発によって1つ又は複数のアミノ酸残基位置で改変された変異ポリペプチド基質種とのライブラリーを含むこと;第1の複数の組換え一倍体酵母細胞と第2の複数の組換え一倍体酵母細胞を液体培地中で合わせて、培養物を産生すること;複数のポリペプチドユビキチンリガーゼ種の1つ又は複数と複数のポリペプチド基質種の1つ又は複数との間の1つ又は複数の相互作用が、第1の複数の組換え一倍体酵母細胞の1つ又は複数と第2の複数の組換え一倍体酵母細胞の1つ又は複数との間の1つ又は複数の交配事象を媒介するような条件下で一定期間、培養物を増殖させて、1つ又は複数の二倍体酵母細胞を産生すること;培養物中の交配事象の数に基づいて、複数のポリペプチドユビキチンリガーゼ種の1つ又は複数と複数のポリペプチド基質種の1つ又は複数との間の相互作用の強さを決定すること;及びポリペプチドの対を同定することであって、ポリペプチドユビキチンリガーゼ種の1つ及びポリペプチド基質種の1つのうち1つ又は両方が、変異誘発によって1つ又は複数のアミノ酸残基位置で修飾されており、ポリペプチドユビキチンリガーゼ種とポリペプチド基質種との間の相互作用の強さ(KD)が、対応する野生型ポリペプチド種間の相互作用よりも少なくとも10%強い又は弱いことを含む。
[0017] 本明細書に組み込まれ、その一部を構成する添付の図面は、1つ又は複数の実施形態を示し、記述とともにこれらの実施形態を説明する。添付の図面は、必ずしも一定の縮尺で描かれているわけではない。添付のグラフ及び図面に示されている値寸法は、説明のみを目的としており、実際の値又は好ましい値又は寸法を表している場合も表していない場合もある。該当する場合、基礎となる特徴の説明を補助するために、一部又は全ての特徴が図示されていない場合がある。
[0043] 添付の図面に関連して以下に記載される説明は、開示された主題の様々な例示的な実施形態の説明であることを意図している。特定の特徴及び機能は、各例示的実施形態に関連して説明される。しかし、当業者には、開示された実施形態がそれらの特定の各特徴及び機能なしで実施され得ることが明らかであろう。
Claims (20)
- a)第1の培養物において、一倍体酵母細胞の表面に、
i)1つ又は複数の野生型標的化タンパク質;及び/又は
ii)前記野生型標的化タンパク質とは少なくとも1つのアミノ酸が異なる1つ又は複数の修飾された標的化タンパク質
の1つ又は両方を発現させること;
b)第2の培養物において、一倍体酵母細胞の表面に、
i)1つ又は複数の野生型標的タンパク質;及び/又は
ii)前記野生型標的タンパク質とは少なくとも1つのアミノ酸が異なる1つ又は複数の修飾された標的タンパク質
の1つ又は両方を発現させること;
c)標的化タンパク質と標的タンパク質との結合が二倍体酵母細胞の形成をもたらすように、前記一倍体酵母細胞の第1の培養物と一倍体酵母細胞の第2の培養物を単一液体培養物に合わせること;及び
d)前記第1及び第2の培養物の前記一倍体酵母細胞間の交配事象の数に基づいて、前記野生型標的タンパク質と前記野生型標的化タンパク質との間の相互作用よりも少なくとも10倍小さい、前記標的化タンパク質と前記標的タンパク質との間の解離定数を生じる、前記標的化タンパク質及び/又は標的タンパク質、又は前記標的化タンパク質及び前記標的タンパク質の両方の、アミノ酸配列変異を決定すること
を含む方法であって、
前記野生型標的タンパク質と前記野生型標的化タンパク質との間の相互作用が、100nMより大きい解離定数(Kd)を有し;
前記標的化タンパク質又は標的タンパク質のいずれかがユビキチンリガーゼであり、
少なくとも1つの前記第1の培養物又は前記第2の培養物が、それぞれ、1つ又は複数の修飾された標的化タンパク質又は標的タンパク質を発現する一倍体酵母細胞を含む、
方法。 - 前記修飾された標的タンパク質及び/又は標的化タンパク質の1つ又は複数が、それぞれの野生型標的タンパク質又は野生型標的化タンパク質の切断型バージョンである、請求項1に記載の方法。
- 前記1つ又は複数の変異が、標的化、ランダム、又は部位飽和変異誘発によって生成される、請求項1に記載の方法。
- 前記修飾された標的化タンパク質と前記標的タンパク質との間の弱い又は強い相互作用が、前記野生型標的化タンパク質と野生型標的タンパク質との間の相互作用よりも約20、30、40、50、100、500、1000パーセント弱いか又は強いと決定される、請求項1に記載の方法。
- 前記弱い相互作用が、1マイクロモーラー又は10マイクロモーラーより大きい解離定数(Kd)を有する、請求項4に記載の方法。
- 前記強い相互作用が、10ナノモーラー又は1ナノモーラー未満の解離定数(Kd)を有する、請求項4に記載の方法。
- 前記弱い相互作用と1つ又は複数の強い相互作用との間の差が、1、2、又は3桁以上である、請求項4に記載の方法。
- 前記ユビキチンリガーゼが、E3ユビキチンリガーゼである、請求項1に記載の方法。
- 1つ又は複数の前記変異が、標的化タンパク質及び/又は標的タンパク質に立体バルクを付加する、請求項1に記載の方法。
- 前記強い相互作用を構成する2つのタンパク質間の界面の構造を決定することをさらに含む、請求項4に記載の方法。
- 前記構造が、結晶学を使用して決定される、請求項10に記載の方法。
- 前記構造が、続いてコンピューターモデリングを用いて決定される、請求項11に記載の方法。
- 前記弱い相互作用の前記構造が、その後、前記強い相互作用の前記構造を取得し、全てのアミノ酸を前記野生型標的タンパク質及び前記野生型標的化タンパク質に見出されるものにコンピューターで復元することによって予測される、請求項10に記載の方法。
- 前記弱い相互作用を安定化させる小分子化合物を同定することをさらに含む、請求項11に記載の方法。
- 前記一倍体酵母細胞の表面上で前記タンパク質を発現させるためにポリヌクレオチドライブラリーを使用することをさらに含み、前記ポリヌクレオチドライブラリーが、10,000個までの標的化タンパク質及び10,000個までの標的タンパク質をコードする、請求項1に記載の方法。
- 前記一倍体酵母細胞の第1及び第2の培養物がそれぞれ、いかなる天然の有性凝集プロセスによっても交配することができない、Mat a細胞及びMat α細胞である、請求項1に記載の方法。
- 決定ステップにおける交配事象の数が、次世代DNA配列決定によって測定される、請求項1に記載の方法。
- 前記ユビキチンリガーゼが、E3ユビキチンリガーゼである、請求項11に記載の方法。
- 前記ユビキチンリガーゼが、E3ユビキチンリガーゼである、請求項15に記載の方法。
- 前記ユビキチンリガーゼが、E3ユビキチンリガーゼである、請求項16に記載の方法。
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US6696251B1 (en) | 1996-05-31 | 2004-02-24 | Board Of Trustees Of The University Of Illinois | Yeast cell surface display of proteins and uses thereof |
US6699658B1 (en) | 1996-05-31 | 2004-03-02 | Board Of Trustees Of The University Of Illinois | Yeast cell surface display of proteins and uses thereof |
US6083693A (en) | 1996-06-14 | 2000-07-04 | Curagen Corporation | Identification and comparison of protein-protein interactions that occur in populations |
US6187535B1 (en) | 1998-02-18 | 2001-02-13 | Institut Pasteur | Fast and exhaustive method for selecting a prey polypeptide interacting with a bait polypeptide of interest: application to the construction of maps of interactors polypeptides |
US6440696B1 (en) | 1999-07-28 | 2002-08-27 | New England Medical Center | E6 targeted protein (E6TP1) |
US7223533B2 (en) | 1999-09-10 | 2007-05-29 | Cadus Technologies, Inc. | Cell surface proteins and use thereof as indicators of activation of cellular signal transduction pathways |
US20020055110A1 (en) | 2000-06-23 | 2002-05-09 | Ian Tomlinson | Matrix screening method |
DK1292710T3 (da) | 2000-06-23 | 2007-12-27 | Domantis Ltd | Fremgangsmåde til matrix-screening |
WO2002064834A1 (en) | 2001-01-04 | 2002-08-22 | Myriad Genetics, Inc. | Novel two-hybrid system and use thereof |
AU2002243477B2 (en) * | 2001-01-05 | 2007-12-20 | New York University | Methods to identify compounds useful for the treatment of proliferative and differentiative disorders |
WO2002086120A1 (fr) | 2001-04-20 | 2002-10-31 | Kansai Chemical Engineering Co., Ltd. | Proteine de liaison aux anticorps et d'expression de levure sur couche superficielle cellulaire et utilisation de ladite proteine |
US6841352B2 (en) * | 2001-06-29 | 2005-01-11 | Myriad Genetics, Inc. | Mating-based method for detecting protein—protein interaction |
EP1438400B1 (en) | 2001-10-01 | 2009-06-17 | Dyax Corp. | Multi-chain eukaryotic display vectors and uses thereof |
US20050181453A1 (en) * | 2003-10-31 | 2005-08-18 | Proteologics, Inc. | Ubiquitin-based protein interaction assays and related compositions |
EP1677113A1 (en) | 2004-12-29 | 2006-07-05 | Max-Delbrück-Centrum für Molekulare Medizin (MDC) | Method for the identification of protein-protein interactions in disease related protein networks |
EP1963538A2 (en) | 2005-11-30 | 2008-09-03 | Guild Associates, Inc. | Differentially fluorescent yeast biosensors for the detection and biodegradation of chemical agents |
US8022188B2 (en) | 2006-04-24 | 2011-09-20 | Abbott Laboratories | Immunosuppressant binding antibodies and methods of obtaining and using same |
US8183030B2 (en) | 2006-10-13 | 2012-05-22 | Archer Daniels Midland Company | Use of cell surface displays in yeast cell catalyst supports |
US20100075326A1 (en) | 2008-09-12 | 2010-03-25 | Cornell University | Yeast surface two-hybrid system for quantitative detection of protein-protein interactions |
US20130096281A1 (en) * | 2010-01-21 | 2013-04-18 | Oxyrane Uk Limited | Methods and compositions for displaying a polypeptide on a yeast cell surface |
EP2530731B1 (en) | 2010-01-25 | 2016-03-30 | LG Chem, Ltd. | Sheet for photovoltaic cells |
WO2011092233A1 (en) | 2010-01-29 | 2011-08-04 | Novartis Ag | Yeast mating to produce high-affinity combinations of fibronectin-based binders |
EP2436766A1 (en) | 2010-09-29 | 2012-04-04 | Deutsches Krebsforschungszentrum | Means and methods for improved protein interaction screening |
US9249410B2 (en) | 2010-12-15 | 2016-02-02 | The Johns Hopkins University | Two-hybrid based screen to identify disruptive residues at multiple protein interfaces |
US20130085072A1 (en) | 2011-10-03 | 2013-04-04 | Los Alamos National Security, Llc | Recombinant renewable polyclonal antibodies |
WO2014144495A1 (en) | 2013-03-15 | 2014-09-18 | Abvitro, Inc. | Single cell bar-coding for antibody discovery |
US10988759B2 (en) | 2016-01-15 | 2021-04-27 | University Of Washington | High throughput protein-protein interaction screening in yeast liquid culture |
US20170205421A1 (en) | 2016-01-15 | 2017-07-20 | University Of Washington | Synthetic yeast agglutination |
US10017758B1 (en) * | 2017-05-25 | 2018-07-10 | Verily Life Sciences Llc | Protein-protein interaction guided mating of yeast |
CN112005115A (zh) | 2018-02-12 | 2020-11-27 | 10X基因组学有限公司 | 表征来自单个细胞或细胞群体的多种分析物的方法 |
US20210147831A1 (en) | 2018-04-27 | 2021-05-20 | The Broad Institute, Inc. | Sequencing-based proteomics |
SG11202102744PA (en) * | 2018-10-16 | 2021-04-29 | Cemm Forschungszentrum Fuer Molekulare Medizin Gmbh | Method for identifying a chemical compound or agent inducing ubiquitination of a protein of interest |
CN110835641B (zh) * | 2019-10-23 | 2023-04-07 | 中山大学肿瘤防治中心(中山大学附属肿瘤医院、中山大学肿瘤研究所) | 基于nanoBRET的促蛋白泛素化降解药物筛选***及方法 |
CN115552003A (zh) | 2020-05-11 | 2022-12-30 | A-阿尔法生物股份有限公司 | 鉴定小分子靶标的高通量筛选方法 |
EP4158015A1 (en) | 2020-06-01 | 2023-04-05 | A-Alpha Bio, Inc. | Methods for characterizing and engineering protein-protein interactions |
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Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2009100746A (ja) | 1998-08-28 | 2009-05-14 | Univ New York | 治療ターゲットとしての新規ユビキチンリガーゼ |
Non-Patent Citations (1)
Title |
---|
PNAS,2017年,Vol.114, No.46,pp.12166-12171 |
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