JP2011501951A5 - - Google Patents
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- JP2011501951A5 JP2011501951A5 JP2010531076A JP2010531076A JP2011501951A5 JP 2011501951 A5 JP2011501951 A5 JP 2011501951A5 JP 2010531076 A JP2010531076 A JP 2010531076A JP 2010531076 A JP2010531076 A JP 2010531076A JP 2011501951 A5 JP2011501951 A5 JP 2011501951A5
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- 235000001014 amino acid Nutrition 0.000 claims 15
- 150000001413 amino acids Chemical class 0.000 claims 14
- 239000000427 antigen Substances 0.000 claims 10
- 102000038129 antigens Human genes 0.000 claims 10
- 108091007172 antigens Proteins 0.000 claims 10
- DHMQDGOQFOQNFH-UHFFFAOYSA-N glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims 10
- 229920001184 polypeptide Polymers 0.000 claims 6
- 239000004471 Glycine Substances 0.000 claims 5
- 229960001230 Asparagine Drugs 0.000 claims 4
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims 4
- 239000004472 Lysine Substances 0.000 claims 4
- 235000009582 asparagine Nutrition 0.000 claims 4
- 230000027455 binding Effects 0.000 claims 4
- MTCFGRXMJLQNBG-UWTATZPHSA-N D-serine Chemical compound OC[C@@H](N)C(O)=O MTCFGRXMJLQNBG-UWTATZPHSA-N 0.000 claims 3
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims 3
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims 3
- 239000004473 Threonine Substances 0.000 claims 3
- 239000003446 ligand Substances 0.000 claims 3
- 108020004707 nucleic acids Proteins 0.000 claims 3
- 150000007523 nucleic acids Chemical class 0.000 claims 3
- 102000004169 proteins and genes Human genes 0.000 claims 3
- 108090000623 proteins and genes Proteins 0.000 claims 3
- 102000008100 Human Serum Albumin Human genes 0.000 claims 2
- 108091006822 Human Serum Albumin Proteins 0.000 claims 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims 2
- 239000002202 Polyethylene glycol Substances 0.000 claims 2
- 235000004279 alanine Nutrition 0.000 claims 2
- 210000004027 cells Anatomy 0.000 claims 2
- 230000000875 corresponding Effects 0.000 claims 2
- 239000003814 drug Substances 0.000 claims 2
- 229940079593 drugs Drugs 0.000 claims 2
- 229920001223 polyethylene glycol Polymers 0.000 claims 2
- 235000018102 proteins Nutrition 0.000 claims 2
- 238000006467 substitution reaction Methods 0.000 claims 2
- 241000894006 Bacteria Species 0.000 claims 1
- 229960002989 Glutamic Acid Drugs 0.000 claims 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims 1
- 102000019040 Nuclear Antigens Human genes 0.000 claims 1
- 108010051791 Nuclear Antigens Proteins 0.000 claims 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 241000700605 Viruses Species 0.000 claims 1
- 102000004965 antibodies Human genes 0.000 claims 1
- 108090001123 antibodies Proteins 0.000 claims 1
- 230000001580 bacterial Effects 0.000 claims 1
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims 1
- 229960002685 biotin Drugs 0.000 claims 1
- 235000020958 biotin Nutrition 0.000 claims 1
- 239000011616 biotin Substances 0.000 claims 1
- 201000011510 cancer Diseases 0.000 claims 1
- 239000002254 cytotoxic agent Substances 0.000 claims 1
- 231100000599 cytotoxic agent Toxicity 0.000 claims 1
- 238000000113 differential scanning calorimetry Methods 0.000 claims 1
- 238000006471 dimerization reaction Methods 0.000 claims 1
- 150000002019 disulfides Chemical class 0.000 claims 1
- 235000013922 glutamic acid Nutrition 0.000 claims 1
- 239000004220 glutamic acid Substances 0.000 claims 1
- 239000012216 imaging agent Substances 0.000 claims 1
- 230000003834 intracellular Effects 0.000 claims 1
- 238000002844 melting Methods 0.000 claims 1
- 230000002285 radioactive Effects 0.000 claims 1
- 239000000126 substance Substances 0.000 claims 1
- 241001515965 unidentified phage Species 0.000 claims 1
- 230000003612 virological Effects 0.000 claims 1
Claims (23)
- 組換えポリペプチド足場であって、
a. A、B、C、D、E、FおよびGと命名された7個のβ鎖ドメイン；
b. 6個のループ領域に連結され、1個のループ領域がそれぞれのβ鎖を接続し、AB、BC、CD、DE、EF、およびFGループと命名された、前記7個のβ鎖ドメイン、
を含み、
c. 少なくとも1個のループ領域が配列番号65〜67のいずれかにおける同族ループ領域の非天然変異体であり、且つ
d. 少なくとも1個のβ鎖ドメインが配列番号65〜67のいずれかにおける同族β鎖ドメインに対して少なくとも90％、少なくとも95％もしくは少なくとも99％の相同性を有し、
Fβ鎖とGβ鎖間；および/またはCβ鎖とFβ鎖間の連結を形成する少なくとも1個のジスルフィド結合を含み、標的に結合する、
前記足場。 - 前記β鎖ドメインが、配列番号65〜67のいずれかにおける同族β鎖ドメインを含む、請求項１に記載の足場。
- 組換えポリペプチド足場であって、
a. A、B、C、D、E、FおよびGと命名された7個のβ鎖ドメイン；
b. 6個のループ領域に連結され、1個のループ領域がそれぞれのβ鎖を接続し、AB、BC、CD、DE、EF、およびFGループと命名された、前記7個のβ鎖ドメイン、
を含み、
c. 少なくとも1個のループ領域が配列番号1、24または61のいずれかにおける同族ループ領域の非天然変異体であり、且つ
d. 少なくとも1個のβ鎖ドメインが配列番号1、24または61のいずれかにおける同族β鎖ドメインに対して少なくとも90％、少なくとも95％もしくは少なくとも99％の相同性を有し、
標的に結合する、
前記足場。 - 前記β鎖ドメインが、配列番号1、24または61のいずれかにおける同族β鎖ドメインを含む、請求項３に記載の足場。
- 前記β鎖ドメインの少なくとも1個が、
Aβ鎖(配列番号235)、Bβ鎖(配列番号229)、Cβ鎖(配列番号230)、Dβ鎖(配列番号236)、Eβ鎖(配列番号232)、Fβ鎖(配列番号237)およびGβ鎖(配列番号234)を含む群より選択される、請求項１または３に記載の足場。 - 前記BCループが、
a. S-X-a-X-b-X-X-X-G(式中、Xは任意のアミノ酸であり、(a)はプロリンまたはアラニンであり、(b)はアラニンまたはグリシンである)の配列を有する9個のアミノ酸；または
b. S-P-c-X-X-X-X-X-X-T-G(式中、Xは任意のアミノ酸であり、(c)はプロリン、セリンまたはグリシンである)の配列を有する11個のアミノ酸；または
c. A-d-P-X-X-X-e-f-X-I-X-G(式中、Xは任意のアミノ酸であり、(d)はプロリン、グルタミン酸またはリジンであり、(e)はアスパラギンまたはグリシンであり、(f)はセリンまたはグリシンである)の配列を有する12個のアミノ酸
を含む、請求項１、２、３、４または５に記載の足場。 - 前記FGループが、
a. X-a-X-X-G-X-X-X-S(式中、Xは任意のアミノ酸であり、(a)はアスパラギン、トレオニンまたはリジンである)の配列を有する9個のアミノ酸；または
b. X-a-X-X-X-X-b-N-P-A(式中、Xは任意のアミノ酸であり、(a)はアスパラギン、トレオニンまたはリジンであり、(b)はセリンまたはグリシンである)の配列を有する10個のアミノ酸；または
c. X-a-X-X-G-X-X-S-N-P-A(式中、Xは任意のアミノ酸であり、(a)はアスパラギン、トレオニンまたはリジンである)の配列を有する11個のアミノ酸
を含む、請求項１、２、３、４、５または６に記載の足場。 - 前記標的が、細胞表面抗原、可溶性抗原、固定化抗原、免疫サイレント抗原、細胞内抗原、核内抗原、自己抗原、非自己抗原、癌抗原、細菌抗原、またはウイルス抗原である、請求項１〜７のいずれか1項に記載の足場。
- 前記足場が、示差走査熱量測定法(DSC)による測定で少なくとも40℃の融点(Tm)を示す、請求項１〜８のいずれか1項に記載の足場。
- 前記足場が異種薬剤にコンジュゲートされたものであり、該薬剤が別の足場、ポリエチレングリコール(PEG)、ヒト血清アルブミン(HSA)、抗体のFc領域、IgG分子、結合ペプチド、細胞傷害剤、放射性標識、画像化剤、His-タグ、ビオチン、Flag-タグ、核酸、またはサイトカインからなる群より選択される、請求項１〜９のいずれか1項に記載の足場。
- 請求項１〜１０のいずれか1項に記載の少なくとも2個の足場を含み、該足場が、別の足場、IgG分子もしくはその断片、Fc領域、二量体化ドメイン、化学的架橋、ジスルフィド結合、またはアミノ酸リンカーにより連結される、多量体足場。
- 請求項１〜１１のいずれか1項に記載の足場をコードする単離された核酸分子。
- 請求項１２に記載の核酸に機能し得る形で連結された発現ベクター。
- 請求項１３に記載のベクターを含む宿主細胞。
- 請求項１〜８のいずれか1項に記載の足場を含み、少なくとも1個のループ領域が、長さおよび/または多様性について無作為化されている、ポリペプチドディスプレイライブラリー。
- 前記足場が、天然タンパク質配列中の対応するループ配列からの少なくとも1個のアミノ酸の欠失、置換または付加により変化する少なくとも2個のループ領域配列を含む、請求項１５に記載のライブラリー。
- 前記2個のループ領域配列が、BC/DE、BC/FG、DE/FG、AB/CD、AB/EF、およびCD/EDループからなる群より選択される、請求項１５に記載のライブラリー。
- 前記足場が、天然タンパク質配列中の対応するループ配列からの少なくとも1個のアミノ酸の欠失、置換または付加により変化する少なくとも3個のループ領域配列を含む、請求項１５に記載のライブラリー。
- 前記足場がリボソーム、バクテリオファージ、ウイルス、細菌、または酵母の表面上にディスプレイされる、請求項１５、１６、１７または１８に記載のポリペプチドディスプレイライブラリー。
- (a)足場：標的リガンド複合体を形成させる条件下で、標的リガンドと請求項１５、１６、１７、１８または１９に記載のライブラリーとを接触させること、および(b)該複合体から、標的リガンドに結合する足場を取得することを含む、標的に結合するポリペプチド足場を取得する方法。
- 工程(b)で得られたタンパク質の足場の少なくとも1個のループを無作為化して、さらに無作為化された足場を作製すること、ならびにさらに無作為化された足場を用いて、工程(a)および(b)を反復することをさらに含む、請求項２０に記載の方法。
- 請求項２０または２１に記載の方法によって取得されるポリペプチド足場。
- 請求項１〜１１のいずれか1項に記載の足場を含む、滅菌された、発熱物質を含まない組成物。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US98420907P | 2007-10-31 | 2007-10-31 | |
| PCT/US2008/012398 WO2009058379A2 (en) | 2007-10-31 | 2008-10-31 | Protein scaffolds |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2014243591A Division JP6034846B2 (ja) | 2007-10-31 | 2014-12-02 | タンパク質足場 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2011501951A JP2011501951A (ja) | 2011-01-20 |
| JP2011501951A5 true JP2011501951A5 (ja) | 2011-12-22 |
Family
ID=40591700
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2010531076A Pending JP2011501951A (ja) | 2007-10-31 | 2008-10-31 | タンパク質足場 |
| JP2014243591A Active JP6034846B2 (ja) | 2007-10-31 | 2014-12-02 | タンパク質足場 |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2014243591A Active JP6034846B2 (ja) | 2007-10-31 | 2014-12-02 | タンパク質足場 |
Country Status (13)
| Country | Link |
|---|---|
| US (2) | US8633297B2 (ja) |
| EP (1) | EP2215246B1 (ja) |
| JP (2) | JP2011501951A (ja) |
| KR (1) | KR101686247B1 (ja) |
| CN (1) | CN101932720A (ja) |
| AU (1) | AU2008319298B2 (ja) |
| BR (1) | BRPI0818810A2 (ja) |
| CA (1) | CA2704229C (ja) |
| ES (1) | ES2533874T3 (ja) |
| HK (1) | HK1147286A1 (ja) |
| MX (1) | MX2010004374A (ja) |
| RU (1) | RU2010121967A (ja) |
| WO (1) | WO2009058379A2 (ja) |
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