JP2013537400A5 - - Google Patents
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- JP2013537400A5 JP2013537400A5 JP2013513670A JP2013513670A JP2013537400A5 JP 2013537400 A5 JP2013537400 A5 JP 2013537400A5 JP 2013513670 A JP2013513670 A JP 2013513670A JP 2013513670 A JP2013513670 A JP 2013513670A JP 2013537400 A5 JP2013537400 A5 JP 2013537400A5
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- 125000000998 L-alanino group Chemical group [H]N([*])[C@](C([H])([H])[H])([H])C(=O)O[H] 0.000 claims 28
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical group NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims 25
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical group CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims 21
- 102000003752 Lipocalin 1 Human genes 0.000 claims 18
- 108010057281 Lipocalin 1 Proteins 0.000 claims 18
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical group OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims 14
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical group OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims 14
- 125000000174 L-prolyl group Chemical group [H]N1C([H])([H])C([H])([H])C([H])([H])[C@@]1([H])C(*)=O 0.000 claims 12
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical group OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims 9
- 125000000510 L-tryptophano group Chemical group [H]C1=C([H])C([H])=C2N([H])C([H])=C(C([H])([H])[C@@]([H])(C(O[H])=O)N([H])[*])C2=C1[H] 0.000 claims 9
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims 8
- 102000010787 Interleukin-4 Receptors Human genes 0.000 claims 6
- 108010038486 Interleukin-4 Receptors Proteins 0.000 claims 6
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 claims 6
- MTCFGRXMJLQNBG-REOHCLBHSA-N L-serine Chemical group OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims 5
- 101710017807 his-44 Proteins 0.000 claims 5
- 150000007523 nucleic acids Chemical class 0.000 claims 5
- 108020004707 nucleic acids Proteins 0.000 claims 5
- 229920001184 polypeptide Polymers 0.000 claims 4
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 3
- 150000001413 amino acids Chemical class 0.000 claims 3
- 230000002163 immunogen Effects 0.000 claims 3
- 101710017531 H4C15 Proteins 0.000 claims 2
- -1 His Chemical compound 0.000 claims 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims 2
- 125000000539 amino acid group Chemical group 0.000 claims 2
- 230000036947 Dissociation constant Effects 0.000 claims 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims 1
- 125000003338 L-glutaminyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C([H])([H])C(=O)N([H])[H] 0.000 claims 1
- 125000003290 L-leucino group Chemical group [H]OC(=O)[C@@]([H])(N([H])[*])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] 0.000 claims 1
- 125000002041 N(2)-L-arginino group Chemical group [H]OC(=O)[C@@]([H])(N([H])[*])C([H])([H])C([H])([H])C([H])([H])N([H])C(=N[H])N([H])[H] 0.000 claims 1
- 125000000534 N(2)-L-lysino group Chemical group [H]OC(=O)[C@@]([H])(N([H])[*])C([H])([H])C([H])([H])C(C([H])([H])N([H])[H])([H])[H] 0.000 claims 1
- 229920001850 Nucleic acid sequence Polymers 0.000 claims 1
- 230000002860 competitive Effects 0.000 claims 1
- 230000000875 corresponding Effects 0.000 claims 1
- 101700022040 his-42 Proteins 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000002703 mutagenesis Methods 0.000 claims 1
- 231100000350 mutagenesis Toxicity 0.000 claims 1
- 238000006467 substitution reaction Methods 0.000 claims 1
Claims (16)
- IL−4レセプターαに対する結合親和性を有するヒト涙液リポカリンのムテインであって、
26の位置におけるPro, Leu, Asn, Tyr, Lys若しくはGlu;32の位置におけるLys;34の位置におけるTrp, Asn, Val, His若しくはArg;56の位置におけるLeu,Ala, His, Lys若しくはMet;58の位置におけるLeu;または、63の位置におけるLys,Ser, Gln若しくはProからなる群から選択される成熟ヒト涙リポカリンのアミノ酸pに対応する位置の少なくとも1つの置換を特徴とするヒト涙液リポカリンのムテイン。 - (i)成熟ヒト涙液リポカリンの直鎖状ポリペプチド配列の27, 28, 30, 31,33, 53, 57, 61, 64, 66, 80, 83, 104〜106及び108の配列位置の1つ又はそれ以上に変異されたアミノ酸残基を含み、
(ii)前記成熟ヒト涙液リポカリンの直鎖状ポリペプチド配列の26, 32, 34, 55,56, 58及び63の配列位置の2つ又はそれ以上に変異されたアミノ酸残基を含み、
前記ムテインのアミノ酸配列は、
(1)Ser 26, Glu34, Leu 55, Lys 58,
(2)Ser 26, Asn34, Ala 55, Lys 58,
(3)Ser 26, Val34,
(4)Pro 26, Ser34,
(5)Pro 26, Ala 55,
(6)Leu 26, Trp 34, Ala 55,
(7)Leu 26, Trp 34, Ile 58,
(8)Asn 26, Asp34,
(9)Asn 26, Ala55,
(10)Tyr 26, His 34, Ala 55,
(11)Tyr 26, His 34, Ala 58,
(12)Lys 26, Arg 34, Ala 55,
(13)Lys 26, Arg 34, Asn 58,
(14)Glu 26, Gly 34, Ala 55,
(15)Glu 26, Gly 34, Leu 58,
のアミノ酸の組み合わせのいずれか1つを含む請求項1に記載のヒト涙液リポカリンのムテイン。 - Ala 31, Tyr 33, Trp 61, Ser 80, Leu 104, Pro 106及びGln 108の少なくとも1つをさらに含む請求項1又は2に記載のヒト涙液リポカリンのムテイン。
- Phe 53, Leu 53, Tyr 64, Met 64, Leu 66及びAsp66の少なくとも1つをさらに含む請求項1又は2に記載のヒト涙液リポカリンのムテイン。
- Arg 27; Cys 28, Arg 30, Ala 31, Tyr 33, Arg 57, Trp 61, Ser 80, Arg83, Leu 104, Cys 105, Pro 106及びGln 108の少なくとも1つをさらに含む請求項1又は2に記載のヒト涙液リポカリンのムテイン。
- (1)Phe 53, Tyr 64, Leu 66と、
(2)Leu 53, Met 64, Asp 66と、
のうちの一方又は両方の群をさらに含む請求項1〜5に記載のヒト涙液リポカリンのムテイン。 - (1)Ser 26; Tyr 32; Leu 55; Gln 56; Lys 58;
(2)Pro 26; Tyr32; Ser 34; Ala 55; Gln 56; Lys 63;
(3)Leu 26; Phe32; Trp 34; Ala 55; Ile 58; Ser 63;
(4)Ser 26; Tyr32; Val 34; Ala 55; Ala 56; Ile 58; Ser 63;
(5)Ser 26; Val32; Asn 34; Ala 55; Gln 56; Lys 58; Lys 63;
(6)Asn 26; Lys 32; Asp 34; Ala 55; His 56; Arg 58; Gln 63;
(7)Tyr 26; Tyr 32; His 34; Ala 55; His 56; Ala 58; Lys 63;
(8)Lys 26; Tyr 32; Arg 34; Ala 55; Lys 56; Asn 58; Pro 63;及び
(9)Glu 26; His 32; Gly 34; Ala 55; Met 56; Leu 58; Lys 63
のいずれか1つの群を含む請求項1又は2に記載のヒト涙液リポカリンのムテイン。 - SEQ ID Nos:3-11のいずれか1つに示された請求項1又は2に記載のヒト涙液リポカリンのムテイン、又はそれらの断片若しくは変異体のヒト涙液リポカリンのムテイン。
- IL−4レセプターαに結合するヒト涙液リポカリンのムテインを生成する方法であって、
(i)成熟ヒト涙液リポカリンの直鎖状ポリペプチド配列の27, 28, 30, 31, 33, 53, 57, 61, 64, 66, 80, 83, 104〜106及び108のアミノ酸配列位置の1つ又はそれ以上と、
(ii)前記成熟ヒト涙液リポカリンの直鎖状ポリペプチド配列の26, 32, 34, 55,56, 58及び63のアミノ酸配列位置の2つ又はそれ以上とにおいて、
ヒト涙液リポカリンをコードする核酸分子に突然変異誘発させることにより、ヒト涙液リポカリンのムテインをコードする1つ又はそれ以上の核酸を得、
26の位置において突然変異された場合、突然変異体は、Ser, Pro, Leu,Asn, Tyr, Lys又はGluを含み;
32の位置において突然変異された場合、突然変異体は、Tyr, Val, Lys又はHisを含み;
34の位置において突然変異された場合、突然変異体は、Glu, Ser, Trp,Asn, Val, Asp, His, Arg又はGlyを含み;
55の位置において突然変異された場合、突然変異体は、Leu又はAlaを含み;
56の位置において突然変異された場合、突然変異体は、Gln, Leu, Ala,His, Lys又はMetを含み;
58の位置において突然変異された場合、突然変異体は、Lys, Ser, Ile,Arg, Ala, Asn又はLeuを含み;
63の位置において突然変異された場合、突然変異体は、Glu, Lys, Ser, Gln又はProを含むステップ(a)と、
(a)において得られた1つ又はそれ以上の前記ムテインの核酸分子を発現システムで発現することにより、1つ又はそれ以上のムテインを得るステップ(b)と、
(b)において得られた1つ又はそれ以上の前記ムテインを濃縮し、選択及び/又は単離により10nM未満の解離定数でIL−4レセプターαに結合するムテインを選択するステップ(c)とを備えていることを特徴とする方法。 - 前記ステップ(c)は、
IL−4レセプターα又はその免疫原性の断片を準備するステップ(ci)と、
1つ又はそれ以上の前記ムテインと前記IL−4レセプターα又はその免疫原性の断片とを接触させることにより、前記IL−4レセプターα又はその免疫原性の断片と結合親和力を有するムテインとの複合体を形成させるステップ(cii)とを含む請求項9に記載の方法。 - 前記ステップ(c)の前記選択は、競合条件下で行われる請求項9又は10に記載の方法。
- 前記ムテインは、
(1)Ser 26, Glu34, Leu 55, Lys 58,
(2)Ser 26, Asn34, Ala 55, Lys 58,
(3)Ser 26, Val34,
(4)Pro 26, Ser34,
(5)Pro 26, Ala 55,
(6)Leu 26, Trp 34, Ala 55,
(7)Leu 26, Trp 34, Ile 58,
(8)Asn 26, Asp34,
(9)Asn 26, Ala55,
(10)Tyr 26, His 34, Ala 55,
(11)Tyr 26, His 34, Ala 58,
(12)Lys 26, Arg 34, Ala 55,
(13)Lys 26, Arg 34, Asn 58,
(14)Glu 26, Gly 34, Ala 55,
(15)Glu 26, Gly 34, Leu 58
のアミノ酸の組み合わせのうちの1つを含む請求項9〜11のいずれか1項に記載の方法。 - 請求項9〜12のいずれか1項に従って取得可能である、又は、取得されることを特徴とするムテイン。
- 請求項1〜8および13のいずれか1項に記載のムテインをコードするヌクレオチド配列を含むことを特徴とする核酸分子。
- 請求項14に記載の核酸分子を含むことを特徴とする宿主細胞。
- 請求項1〜8及び13のいずれか1項に記載のムテインを含むことを特徴とする医薬又は診断用の組成物。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US35246110P | 2010-06-08 | 2010-06-08 | |
US61/352,461 | 2010-06-08 | ||
PCT/EP2011/059420 WO2011154420A2 (en) | 2010-06-08 | 2011-06-08 | Tear lipocalin muteins binding il-4 r alpha |
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JP2015218174A Division JP2016028604A (ja) | 2010-06-08 | 2015-11-06 | IL−4Rαに結合する涙液リポカリンムテイン |
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JP2013537400A JP2013537400A (ja) | 2013-10-03 |
JP2013537400A5 true JP2013537400A5 (ja) | 2013-11-14 |
JP5902679B2 JP5902679B2 (ja) | 2016-04-13 |
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JP2013513670A Active JP5902679B2 (ja) | 2010-06-08 | 2011-06-08 | IL−4Rαに結合する涙液リポカリンムテイン |
JP2015218174A Pending JP2016028604A (ja) | 2010-06-08 | 2015-11-06 | IL−4Rαに結合する涙液リポカリンムテイン |
JP2019014631A Active JP6766194B2 (ja) | 2010-06-08 | 2019-01-30 | IL−4Rαに結合する涙液リポカリンムテイン |
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JP2015218174A Pending JP2016028604A (ja) | 2010-06-08 | 2015-11-06 | IL−4Rαに結合する涙液リポカリンムテイン |
JP2019014631A Active JP6766194B2 (ja) | 2010-06-08 | 2019-01-30 | IL−4Rαに結合する涙液リポカリンムテイン |
Country Status (22)
Country | Link |
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US (5) | US8986951B2 (ja) |
EP (1) | EP2580236B1 (ja) |
JP (3) | JP5902679B2 (ja) |
CN (3) | CN105949301B (ja) |
AU (1) | AU2011263786B2 (ja) |
CA (1) | CA2800026C (ja) |
CY (1) | CY1122339T1 (ja) |
DK (1) | DK2580236T3 (ja) |
ES (1) | ES2729652T3 (ja) |
HR (1) | HRP20190689T1 (ja) |
HU (1) | HUE043835T2 (ja) |
LT (1) | LT2580236T (ja) |
NZ (1) | NZ603562A (ja) |
PL (1) | PL2580236T3 (ja) |
PT (1) | PT2580236T (ja) |
RS (1) | RS58839B1 (ja) |
RU (1) | RU2569745C2 (ja) |
SG (1) | SG186161A1 (ja) |
SI (1) | SI2580236T1 (ja) |
TR (1) | TR201906295T4 (ja) |
WO (1) | WO2011154420A2 (ja) |
ZA (1) | ZA201209160B (ja) |
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