JPWO2019169212A5 - - Google Patents
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- JPWO2019169212A5 JPWO2019169212A5 JP2020545621A JP2020545621A JPWO2019169212A5 JP WO2019169212 A5 JPWO2019169212 A5 JP WO2019169212A5 JP 2020545621 A JP2020545621 A JP 2020545621A JP 2020545621 A JP2020545621 A JP 2020545621A JP WO2019169212 A5 JPWO2019169212 A5 JP WO2019169212A5
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- 239000000427 antigen Substances 0.000 claims description 104
- 102000038129 antigens Human genes 0.000 claims description 104
- 108091007172 antigens Proteins 0.000 claims description 104
- 102000004965 antibodies Human genes 0.000 claims description 101
- 108090001123 antibodies Proteins 0.000 claims description 101
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 40
- 229920000023 polynucleotide Polymers 0.000 claims description 20
- 239000002157 polynucleotide Substances 0.000 claims description 20
- 108010047041 Complementarity Determining Regions Proteins 0.000 claims description 16
- 230000001225 therapeutic Effects 0.000 claims description 14
- 150000007523 nucleic acids Chemical class 0.000 claims description 13
- 108020004707 nucleic acids Proteins 0.000 claims description 13
- 210000004027 cells Anatomy 0.000 claims description 11
- 238000001514 detection method Methods 0.000 claims description 10
- 201000011510 cancer Diseases 0.000 claims description 8
- 238000003364 immunohistochemistry Methods 0.000 claims description 6
- 206010006187 Breast cancer Diseases 0.000 claims description 5
- HSTOKWSFWGCZMH-UHFFFAOYSA-N 3,3'-Diaminobenzidine Chemical compound C1=C(N)C(N)=CC=C1C1=CC=C(N)C(N)=C1 HSTOKWSFWGCZMH-UHFFFAOYSA-N 0.000 claims description 4
- 238000002965 ELISA Methods 0.000 claims description 4
- 230000035693 Fab Effects 0.000 claims description 4
- 208000002154 Non-Small-Cell Lung Carcinoma Diseases 0.000 claims description 4
- 108009000071 Non-small cell lung cancer Proteins 0.000 claims description 4
- NFGXHKASABOEEW-UHFFFAOYSA-N (+)-methoprene Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 claims description 2
- 206010005003 Bladder cancer Diseases 0.000 claims description 2
- 210000000170 Cell Membrane Anatomy 0.000 claims description 2
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 claims description 2
- 208000006402 Ductal Carcinoma Diseases 0.000 claims description 2
- 206010014733 Endometrial cancer Diseases 0.000 claims description 2
- 206010033128 Ovarian cancer Diseases 0.000 claims description 2
- 208000008443 Pancreatic Carcinoma Diseases 0.000 claims description 2
- 206010038389 Renal cancer Diseases 0.000 claims description 2
- 108010070144 Single-Chain Antibodies Proteins 0.000 claims description 2
- 102000005632 Single-Chain Antibodies Human genes 0.000 claims description 2
- 206010041823 Squamous cell carcinoma Diseases 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 238000004166 bioassay Methods 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 230000001086 cytosolic Effects 0.000 claims description 2
- 230000001809 detectable Effects 0.000 claims description 2
- 201000010982 kidney cancer Diseases 0.000 claims description 2
- 201000002528 pancreatic cancer Diseases 0.000 claims description 2
- 201000002510 thyroid cancer Diseases 0.000 claims description 2
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 2
- 206010003445 Ascites Diseases 0.000 description 1
- 210000002751 Lymph Anatomy 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 210000002381 Plasma Anatomy 0.000 description 1
- 210000002966 Serum Anatomy 0.000 description 1
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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Description
他の実施形態は以下のクレームの範囲内にある。
特定の実施形態では、例えば以下の項目が提供される。
(項目1)
ヒトB7-H4に特異的に結合する単離された抗体またはその抗原結合断片であって、配列番号22のアミノ酸配列を含む重鎖可変領域(VH)相補性決定領域(CDR)1と、配列番号23のアミノ酸配列を含むVH CDR2と、配列番号24のアミノ酸配列を含むVH CDR3と、配列番号25のアミノ酸配列を含む軽鎖可変領域(VL)CDR1と、配列番号26、27または28のアミノ酸配列を含むVL CDR2と、配列番号29のアミノ酸配列を含むVL CDR3とを含む、前記抗体またはその抗原結合断片。
(項目2)
前記抗体またはその抗原結合断片が、配列番号6、7または8のアミノ酸配列を含むVHを含む、項目1に記載の抗体またはその抗原結合断片。
(項目3)
前記抗体またはその抗原結合断片が、配列番号9、10または11のアミノ酸配列を含むVLを含む、項目1または2に記載の抗体またはその抗原結合断片。
(項目4)
ヒトB7-H4に特異的に結合する単離された抗体またはその抗原結合断片であって、前記抗体またはその抗原結合断片が重鎖可変領域及び軽鎖可変領域を含み、前記重鎖可変領域が配列番号6、7または8のアミノ酸配列を含む、前記抗体またはその抗原結合断片。
(項目5)
ヒトB7-H4に特異的に結合する単離された抗体またはその抗原結合断片であって、前記抗体が重鎖可変領域及び軽鎖可変領域を含み、前記軽鎖可変領域が配列番号9、10または11のアミノ酸配列を含む、前記抗体またはその抗原結合断片。
(項目6)
ヒトB7-H4に特異的に結合する単離された抗体またはその抗原結合断片であって、(a)それぞれ配列番号6及び9;
(b)それぞれ配列番号6及び10;
(c)それぞれ配列番号6及び11;
(d)それぞれ配列番号7及び9;
(e)それぞれ配列番号7及び10;
(f)それぞれ配列番号7及び11;
(g)それぞれ配列番号8及び9;
(h)それぞれ配列番号8及び10;または
(i)それぞれ配列番号8及び11のアミノ酸配列を含む重鎖可変領域及び軽鎖可変領域を含む、前記抗体またはその抗原結合断片。
(項目7)
前記抗体またはその抗原結合断片がさらに重鎖定常領域を含む、項目1~6のいずれか1項に記載の抗体またはその抗原結合断片。
(項目8)
前記重鎖定常領域がマウスIgG
1
またはIgG
2a
の重鎖定常領域である、項目7に記載の抗体またはその抗原結合断片。
(項目9)
前記抗体または抗原結合断片がさらに軽鎖定常領域を含む、項目1~8のいずれか1項に記載の抗体またはその抗原結合断片。
(項目10)
前記軽鎖定常領域がマウスIgGκ軽鎖定常領域である、項目9に記載の抗体またはその抗原結合断片。
(項目11)
前記抗体またはその抗原結合断片が配列番号16、17、18、89、90または91のアミノ酸配列を含む重鎖を含む、項目1~6のいずれか1項に記載の抗体またはその抗原結合断片。
(項目12)
前記抗体またはその抗原結合断片が配列番号19、30または31のアミノ酸配列を含む軽鎖を含む、項目1~6または11のいずれか1項に記載の抗体またはその抗原結合断片。
(項目13)
前記抗体または抗原結合断片が、
(a)それぞれ配列番号16及び19;
(b)それぞれ配列番号16及び30;
(c)それぞれ配列番号16及び31;
(d)それぞれ配列番号17及び19;
(e)それぞれ配列番号17及び30;
(f)それぞれ配列番号17及び31;
(g)それぞれ配列番号18及び19;
(h)それぞれ配列番号18及び30;
(i)それぞれ配列番号18及び31;
(j)それぞれ配列番号89及び19;
(k)それぞれ配列番号90及び19;または
(l)それぞれ配列番号91及び19のアミノ酸配列を含む重鎖及び軽鎖を含む、項目1~6のいずれか1項に記載の抗体またはその抗原結合断片。
(項目14)
ヒトB7-H4に特異的に結合する単離された抗体またはその抗原結合断片であって、前記抗体またはその抗原結合断片が、J512、J513、J514、J515、J516、J517、J518、J519、J520、J521、及びJ522から成る群から選択される抗体のVH CDR1、VH CDR2、VH CDR3、VL CDR1、VL CDR2、及びVL CDR3を含む、前記抗体またはその抗原結合断片。
(項目15)
前記CDRがKabatが定義したCDR、Chothiaが定義したCDR、またはAbMの定義のCDRである、項目14に記載の抗体またはその抗原結合断片。
(項目16)
項目1~15のいずれか1項に記載の抗体またはその抗原結合断片と同じヒトB7-H4のエピトープに特異的に結合する、単離された抗体またはその抗原結合断片。
(項目17)
項目1~15のいずれか1項に記載の抗体またはその抗原結合断片のヒトB7-H4への結合を 競合して阻害する、単離された抗体またはその抗原結合断片。
(項目18)
前記抗体またはその抗原結合断片がマウスの抗体またはその抗原結合断片である、項目1~17のいずれか1項に記載の抗体またはその抗原結合断片。
(項目19)
完全長抗体である、項目1~18のいずれか1項に記載の抗体またはその抗原結合断片。
(項目20)
抗原結合断片である、項目1~18のいずれか1項に記載の抗体またはその抗原結合断片。
(項目21)
前記抗原結合断片が、Fab、Fab’、F(ab’)
2
、単鎖Fv(scFv)、ジスルフィド結合Fv、ミニボディ、F(ab’)
3
、ダイアボディ、(scFv)
2
、またはscFv-Fcを含む、項目20に記載の抗原結合断片。
(項目22)
さらに、検出可能な標識を含む、項目1~21のいずれか1項に記載の抗体またはその抗原結合断片。
(項目23)
項目1~22のいずれか1項に記載の抗体またはその抗原結合断片の重鎖可変領域または重鎖をコードする核酸分子を含む、単離されたポリヌクレオチド。
(項目24)
前記核酸分子が配列番号6、7もしくは8のVHをコードする、または配列番号16、17もしくは18の重鎖をコードする項目23に記載の単離されたポリヌクレオチド。
(項目25)
前記核酸分子が配列番号12、13、14、93、94または95の配列を含む、項目24に記載の単離されたポリヌクレオチド。
(項目26)
項目1~22のいずれか1項に記載の抗体またはその抗原結合断片の軽鎖可変領域または軽鎖をコードする核酸分子を含む、単離されたポリヌクレオチド。
(項目27)
前記核酸分子が配列番号9、10もしくは11のVLをコードする、または配列番号19、30もしくは31の軽鎖をコードする、項目26に記載の単離されたポリヌクレオチド。
(項目28)
前記核酸分子が配列番号15、96または97の配列を含む、項目27に記載の単離されたポリヌクレオチド。
(項目29)
項目1~22のいずれか1項に記載の抗体またはその抗原結合断片の重鎖可変領域または重鎖と、項目1~22のいずれか1項に記載の抗体またはその抗原結合断片の軽鎖可変領域または軽鎖とをコードする核酸分子を含む、単離されたポリヌクレオチド。
(項目30)
項目23~29のいずれか1項に記載のポリヌクレオチドを含む、単離されたベクター。
(項目31)
項目23~29のいずれか1項に記載のポリヌクレオチド、項目30に記載のベクター、または項目23~25のいずれか1項に記載のポリヌクレオチドを含む第1のベクターと項目26~28のいずれか1項に記載のポリヌクレオチドを含む第2のベクターを含む、宿主細胞。
(項目32)
CHO細胞またはHEK細胞である、項目31に記載の宿主細胞。
(項目33)
ヒトB7-H4に特異的に結合する抗体またはその抗原結合断片を産生させる方法であって、前記核酸分子が発現され、前記抗体またはその抗原結合断片が産生されるように項目31または32に記載の宿主細胞を培養することを含む、前記方法。
(項目34)
ヒトB7-H4に特異的に結合し、且つ項目23~29のいずれか1項に記載のポリヌクレオチドによってコードされる、単離された抗体またはその抗原結合断片。
(項目35)
試料にてB7-H4を検出する方法であって、前記試料を項目1~22または34のいずれか1項に記載の抗体またはその抗原結合断片と接触させることを含み、任意で、前記方法はさらに、前記抗体またはその抗原結合断片のB7-H4に対する結合を検出することを含む、前記方法。
(項目36)
前記試料が対象におけるがんから得られる項目35に記載の方法。
(項目37)
B7-H4が検出された後、治療用の抗B7-H4抗体またはその抗原結合断片を前記対象に投与することをさらに含む、項目36に記載の方法。
(項目38)
対象にてB7-H4を発現するがんを治療する方法であって、前記方法が治療用の抗B7-H4抗体またはその抗原結合断片を前記対象に投与することを含み、その際、B7-H4の発現が、前記対象から得られた試料にて項目1~21または33のいずれか1項に記載の抗体またはその抗原結合断片を用いて検出された、前記方法。
(項目39)
前記がんから得られた前記試料にて前記B7-H4を検出することをさらに含む、項目38に記載の方法。
(項目40)
前記検出されたB7-H4が細胞膜B7-H4である、項目35~39のいずれか1項に記載の方法。
(項目41)
前記検出されたB7-H4が細胞質B7-H4である、項目35~39のいずれか1項に記載の方法。
(項目42)
B7-H4が細胞全体において検出される、項目35~39のいずれか1項に記載の方法。
(項目43)
前記B7-H4が循環腫瘍細胞で検出される、項目35~42のいずれか1項に記載の方法。
(項目44)
前記試料が、対象に由来する固形組織、生検、腹水、血漿、血清、リンパ液、または腫瘍である、項目35~43のいずれか1項に記載の方法。
(項目45)
前記がんが、乳癌、乳管癌、子宮内膜癌、卵巣癌、非小細胞肺癌、膵臓癌、甲状腺癌、腎臓癌及び膀胱癌から成る群から選択される、項目36~44のいずれか1項に記載の方法。
(項目46)
前記乳癌がトリプルネガティブ乳癌であり、または前記非小細胞肺癌が扁平上皮癌である、項目45に記載の方法。
(項目47)
前記検出方法が、酵素結合免疫吸着アッセイ(ELISA)、蛍光活性化細胞選別(FACS)アッセイ、または免疫組織化学(IHC)を使用する、項目35~46のいずれか1項に記載の方法。
(項目48)
前記検出方法がIHCを使用し、項目1~21または34のいずれか1項に記載の抗体またはその抗原結合断片の濃度が約1~約50μg/mLである、項目47に記載の方法。
(項目49)
前記抗体またはその抗原結合断片の濃度が約1~約20μg/mLである、項目48に記載の方法。
(項目50)
前記抗体またはその抗原結合断片の濃度が約10μg/mLである、項目49に記載の方法。
(項目51)
前記検出方法がIHCを使用し、3,3’-ジアミノベンジジン(DAB)が検出に使用される、項目47~50のいずれか1項に記載の方法。
(項目52)
前記対象がヒトである、項目36~51のいずれか1項に記載の方法。
(項目53)
項目1~21または34のいずれか1項に記載の抗体またはその抗原結合断片と、(a)検出試薬、(b)B7-H4抗原、(c)治療用抗B7-H4抗体、または(d)(c)(a)~(c)の組み合わせのいずれかとを含む、キット。
(項目54)
前記治療用の抗体または抗原結合断片が、20502または22213のVH CDR1、VH CDR2、VH CDR3、VL CDR1、VL CDR2、及びVL CDR3を含む、項目37~53のいずれか1項に記載の方法またはキット。
(項目55)
前記CDRが、Kabatが定義したCDR、Chothiaが定義したCDR、またはAbMの定義のCDRである項目54に記載の方法またはキット。
(項目56)
前記VH CDR1、VH CDR2、VH CDR3、VL CDR1、VL CDR2、及びVL CDR3が、それぞれ配列番号32~37のアミノ酸配列、またはそれぞれ配列番号58~63のアミノ酸配列を含む、項目55に記載の方法またはキット。
(項目57)
前記治療用の抗体または抗原結合断片が、重鎖可変領域及び軽鎖可変領域を含み、前記重鎖可変領域が配列番号54または配列番号64のアミノ酸配列を含む、項目37~56のいずれか1項に記載の方法またはキット。
(項目58)
前記治療用の抗体または抗原結合断片が、重鎖可変領域及び軽鎖可変領域を含み、前記軽鎖可変領域が配列番号55または配列番号65のアミノ酸配列を含む、項目37~56のいずれか1項に記載の方法またはキット。
(項目59)
前記治療用の抗体または抗原結合断片が、(i)配列番号54のアミノ酸配列を含む重鎖可変領域及び配列番号55のアミノ酸配列を含む軽鎖可変領域を含む、または(ii)配列番号64のアミノ酸配列を含む重鎖可変領域及び配列番号65のアミノ酸配列を含む軽鎖可変領域を含む、項目37~56のいずれか1項に記載の方法またはキット。
(項目60)
前記治療用の抗体またはその抗原結合断片が、配列番号56または配列番号74のアミノ酸配列を含む重鎖を含む、項目37~56のいずれか1項に記載の方法またはキット。
(項目61)
前記治療用の抗体またはその抗原結合断片が、配列番号57または配列番号75のアミノ酸配列を含む軽鎖を含む、項目37~56のいずれか1項に記載の方法またはキット。
(項目62)
前記治療用の抗体またはその抗原結合断片が(i)配列番号56のアミノ酸配列を含む重鎖及び配列番号57のアミノ酸配列を含む軽鎖を含む、または(ii)配列番号74のアミノ酸配列を含む重鎖及び配列番号75のアミノ酸配列を含む軽鎖を含む、項目37~56のいずれか1項に記載の方法またはキット。
Other embodiments are within the scope of the following claims.
In certain embodiments, for example, the following items are provided.
(Item 1)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4, the heavy chain variable region (VH) complementarity determining region (CDR) 1 containing the amino acid sequence of SEQ ID NO: 22, and the sequence. VH CDR2 containing the amino acid sequence of SEQ ID NO: 23, VH CDR3 containing the amino acid sequence of SEQ ID NO: 24, light chain variable region (VL) CDR1 containing the amino acid sequence of SEQ ID NO: 25, and amino acids of SEQ ID NO: 26, 27 or 28. The antibody or antigen-binding fragment thereof comprising VL CDR2 comprising a sequence and VL CDR3 comprising the amino acid sequence of SEQ ID NO: 29.
(Item 2)
The antibody or antigen-binding fragment thereof according to item 1, wherein the antibody or antigen-binding fragment thereof comprises VH containing the amino acid sequence of SEQ ID NO: 6, 7 or 8.
(Item 3)
The antibody or antigen-binding fragment thereof according to item 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises VL containing the amino acid sequence of SEQ ID NO: 9, 10 or 11.
(Item 4)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4, wherein the antibody or antigen-binding fragment thereof contains a heavy chain variable region and a light chain variable region, and the heavy chain variable region is contained. The antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO: 6, 7 or 8.
(Item 5)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4, wherein the antibody comprises a heavy chain variable region and a light chain variable region, and the light chain variable region is SEQ ID NO: 9, 10 Or the antibody or antigen-binding fragment thereof comprising 11 amino acid sequences.
(Item 6)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4, (a) SEQ ID NOs: 6 and 9, respectively;
(B) SEQ ID NOs: 6 and 10, respectively;
(C) SEQ ID NOs: 6 and 11;
(D) SEQ ID NOs: 7 and 9, respectively;
(E) SEQ ID NOs: 7 and 10, respectively;
(F) SEQ ID NOs: 7 and 11, respectively;
(G) SEQ ID NOs: 8 and 9, respectively;
(H) SEQ ID NOs: 8 and 10; or
(I) The antibody or antigen-binding fragment thereof, which comprises a heavy chain variable region and a light chain variable region containing the amino acid sequences of SEQ ID NOs: 8 and 11, respectively.
(Item 7)
Item 6. The antibody or antigen-binding fragment thereof according to any one of Items 1 to 6, wherein the antibody or antigen-binding fragment thereof further contains a heavy chain constant region.
(Item 8)
Item 7. The antibody or antigen-binding fragment thereof according to item 7, wherein the heavy chain constant region is a heavy chain constant region of mouse IgG 1 or IgG 2a .
(Item 9)
Item 6. The antibody or antigen-binding fragment thereof according to any one of Items 1 to 8, wherein the antibody or antigen-binding fragment further comprises a light chain constant region.
(Item 10)
Item 9. The antibody or antigen-binding fragment thereof according to Item 9, wherein the light chain constant region is a mouse IgGκ light chain constant region.
(Item 11)
Item 6. The antibody or antigen-binding fragment thereof according to any one of Items 1 to 6, wherein the antibody or an antigen-binding fragment thereof comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 16, 17, 18, 89, 90 or 91.
(Item 12)
Item 6. The antibody or antigen-binding fragment thereof according to any one of Items 1 to 6 or 11, wherein the antibody or antigen-binding fragment thereof comprises a light chain comprising the amino acid sequence of SEQ ID NO: 19, 30 or 31.
(Item 13)
The antibody or antigen-binding fragment
(A) SEQ ID NOs: 16 and 19;
(B) SEQ ID NOs: 16 and 30, respectively;
(C) SEQ ID NOs: 16 and 31, respectively;
(D) SEQ ID NOs: 17 and 19, respectively;
(E) SEQ ID NOs: 17 and 30, respectively;
(F) SEQ ID NOs: 17 and 31, respectively;
(G) SEQ ID NOs: 18 and 19, respectively;
(H) SEQ ID NOs: 18 and 30, respectively;
(I) SEQ ID NOs: 18 and 31, respectively;
(J) SEQ ID NOs: 89 and 19;
(K) SEQ ID NOs: 90 and 19; or
(L) The antibody or antigen-binding fragment thereof according to any one of Items 1 to 6, which comprises a heavy chain and a light chain containing the amino acid sequences of SEQ ID NOs: 91 and 19, respectively.
(Item 14)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4, wherein the antibody or antigen-binding fragment thereof is J512, J513, J514, J515, J516, J517, J518, J518, J520. , J521, and an antigen-binding fragment thereof comprising VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of an antibody selected from the group consisting of J522.
(Item 15)
The antibody or antigen-binding fragment thereof according to item 14, wherein the CDR is a CDR defined by Kabat, a CDR defined by Chothia, or a CDR defined by AbM.
(Item 16)
An isolated antibody or antigen-binding fragment thereof that specifically binds to the same epitope of human B7-H4 as the antibody or antigen-binding fragment thereof according to any one of items 1 to 15.
(Item 17)
An isolated antibody or antigen-binding fragment thereof that competitively inhibits the binding of the antibody or antigen-binding fragment thereof according to any one of items 1 to 15 to human B7-H4.
(Item 18)
Item 6. The antibody or antigen-binding fragment thereof according to any one of Items 1 to 17, wherein the antibody or antigen-binding fragment thereof is a mouse antibody or antigen-binding fragment thereof.
(Item 19)
The antibody or antigen-binding fragment thereof according to any one of items 1 to 18, which is a full-length antibody.
(Item 20)
Item 6. The antibody according to any one of Items 1 to 18, which is an antigen-binding fragment, or an antigen-binding fragment thereof.
(Item 21)
The antigen-binding fragment is Fab, Fab', F (ab') 2 , single chain Fv (scFv), disulfide bond Fv, minibody, F (ab') 3 , diabodies, (scFv) 2 , or scFv-. The antigen-binding fragment according to item 20, which comprises Fc.
(Item 22)
The antibody or antigen-binding fragment thereof according to any one of items 1 to 21, further comprising a detectable label.
(Item 23)
An isolated polynucleotide comprising a heavy chain variable region or a nucleic acid molecule encoding a heavy chain of the antibody according to any one of items 1 to 22 or an antigen-binding fragment thereof.
(Item 24)
23. The isolated polynucleotide according to item 23, wherein the nucleic acid molecule encodes a VH of SEQ ID NO: 6, 7 or 8 or encodes a heavy chain of SEQ ID NO: 16, 17 or 18.
(Item 25)
24. The isolated polynucleotide according to item 24, wherein the nucleic acid molecule comprises the sequence of SEQ ID NO: 12, 13, 14, 93, 94 or 95.
(Item 26)
An isolated polynucleotide comprising a light chain variable region or a nucleic acid molecule encoding a light chain of the antibody according to any one of items 1 to 22 or an antigen-binding fragment thereof.
(Item 27)
26. The isolated polynucleotide according to item 26, wherein the nucleic acid molecule encodes the VL of SEQ ID NO: 9, 10 or 11 or the light chain of SEQ ID NO: 19, 30 or 31.
(Item 28)
27. The isolated polynucleotide according to item 27, wherein the nucleic acid molecule comprises the sequence of SEQ ID NO: 15, 96 or 97.
(Item 29)
The heavy chain variable region or heavy chain of the antibody or antigen-binding fragment thereof according to any one of items 1 to 22, and the light chain variable of the antibody or antigen-binding fragment thereof according to any one of items 1 to 22. An isolated polynucleotide comprising a nucleic acid molecule encoding a region or a light chain.
(Item 30)
An isolated vector comprising the polynucleotide according to any one of items 23 to 29.
(Item 31)
The polynucleotide according to any one of items 23 to 29, the vector according to item 30, or the first vector containing the polynucleotide according to any one of items 23 to 25 and any of items 26 to 28. A host cell comprising a second vector comprising the polynucleotide according to item 1.
(Item 32)
31. The host cell according to item 31, which is a CHO cell or a HEK cell.
(Item 33)
Item 31 or 32, wherein the method for producing an antibody or an antigen-binding fragment thereof that specifically binds to human B7-H4 is such that the nucleic acid molecule is expressed and the antibody or the antigen-binding fragment thereof is produced. The method comprising culturing a host cell of the above.
(Item 34)
An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4 and is encoded by the polynucleotide according to any one of items 23 to 29.
(Item 35)
A method for detecting B7-H4 in a sample, comprising contacting the sample with the antibody or antigen-binding fragment thereof according to any one of items 1 to 22 or 34, optionally said the method. The method further comprises detecting the binding of the antibody or antigen-binding fragment thereof to B7-H4.
(Item 36)
35. The method of item 35, wherein the sample is obtained from cancer in a subject.
(Item 37)
36. The method of item 36, further comprising administering to the subject a therapeutic anti-B7-H4 antibody or antigen-binding fragment thereof after B7-H4 is detected.
(Item 38)
A method of treating a cancer expressing B7-H4 in a subject, wherein the method comprises administering to the subject a therapeutic anti-B7-H4 antibody or antigen-binding fragment thereof, in which case B7- The method as described above, wherein expression of H4 was detected in a sample obtained from the subject using the antibody according to any one of items 1 to 21 or 33 or an antigen-binding fragment thereof.
(Item 39)
38. The method of item 38, further comprising detecting the B7-H4 in the sample obtained from the cancer.
(Item 40)
The method according to any one of items 35 to 39, wherein the detected B7-H4 is a cell membrane B7-H4.
(Item 41)
The method according to any one of items 35 to 39, wherein the detected B7-H4 is cytoplasmic B7-H4.
(Item 42)
The method according to any one of items 35 to 39, wherein B7-H4 is detected in the whole cell.
(Item 43)
The method according to any one of items 35 to 42, wherein the B7-H4 is detected in circulating tumor cells.
(Item 44)
The method according to any one of items 35 to 43, wherein the sample is a solid tissue, biopsy, ascites, plasma, serum, lymph, or tumor derived from the subject.
(Item 45)
Any of items 36 to 44, wherein the cancer is selected from the group consisting of breast cancer, ductal carcinoma, endometrial cancer, ovarian cancer, non-small cell lung cancer, pancreatic cancer, thyroid cancer, kidney cancer and bladder cancer. The method according to item 1.
(Item 46)
45. The method of item 45, wherein the breast cancer is triple negative breast cancer or the non-small cell lung cancer is squamous cell carcinoma.
(Item 47)
Item 6. The method according to any one of items 35 to 46, wherein the detection method uses an enzyme-linked immunosorbent assay (ELISA), a fluorescence activated cell selection (FACS) assay, or immunohistochemistry (IHC).
(Item 48)
47. The method of item 47, wherein the detection method uses IHC and the concentration of the antibody or antigen-binding fragment thereof according to any one of items 1 to 21 or 34 is about 1 to about 50 μg / mL.
(Item 49)
48. The method of item 48, wherein the concentration of the antibody or antigen-binding fragment thereof is from about 1 to about 20 μg / mL.
(Item 50)
49. The method of item 49, wherein the concentration of the antibody or antigen-binding fragment thereof is about 10 μg / mL.
(Item 51)
The method according to any one of items 47 to 50, wherein the detection method uses IHC and 3,3'-diaminobenzidine (DAB) is used for detection.
(Item 52)
The method according to any one of items 36 to 51, wherein the subject is a human.
(Item 53)
The antibody or antigen-binding fragment thereof according to any one of items 1 to 21 or 34, and (a) detection reagent, (b) B7-H4 antigen, (c) therapeutic anti-B7-H4 antibody, or (d). ) (C) A kit comprising any of the combinations (a) to (c).
(Item 54)
38. The method of any one of items 37-53, wherein the therapeutic antibody or antigen binding fragment comprises 20502 or 22213 VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3. kit.
(Item 55)
54. The method or kit of item 54, wherein the CDR is a Kabat-defined CDR, a Chothia-defined CDR, or an AbM-defined CDR.
(Item 56)
55. The method of item 55, wherein the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 each contain the amino acid sequences of SEQ ID NOs: 32 to 37, or the amino acid sequences of SEQ ID NOs: 58 to 63, respectively. Or a kit.
(Item 57)
One of items 37-56, wherein the therapeutic antibody or antigen binding fragment comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54 or SEQ ID NO: 64. The method or kit described in the section.
(Item 58)
One of items 37-56, wherein the therapeutic antibody or antigen binding fragment comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 55 or SEQ ID NO: 65. The method or kit described in the section.
(Item 59)
The therapeutic antibody or antigen binding fragment comprises (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 55, or (ii) of SEQ ID NO: 64. The method or kit according to any one of items 37 to 56, comprising a heavy chain variable region comprising an amino acid sequence and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 65.
(Item 60)
37. The method or kit of item 37-56, wherein the therapeutic antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 74.
(Item 61)
37. The method or kit of any one of items 37-56, wherein the therapeutic antibody or antigen binding fragment thereof comprises a light chain comprising the amino acid sequence of SEQ ID NO: 57 or SEQ ID NO: 75.
(Item 62)
The therapeutic antibody or antigen-binding fragment thereof comprises (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 56 and a light chain comprising the amino acid sequence of SEQ ID NO: 57, or (ii) comprising the amino acid sequence of SEQ ID NO: 74. 37. The method or kit of any one of items 37-56, comprising a heavy chain and a light chain comprising the amino acid sequence of SEQ ID NO: 75.
Claims (28)
(a)それぞれ配列番号6及び9;
(b)それぞれ配列番号6及び10;
(c)それぞれ配列番号6及び11;
(d)それぞれ配列番号7及び9;
(e)それぞれ配列番号7及び10;
(f)それぞれ配列番号7及び11;
(g)それぞれ配列番号8及び9;
(h)それぞれ配列番号8及び10;または
(i)それぞれ配列番号8及び11のアミノ酸配列を含む重鎖可変領域及び軽鎖可変領域を含む、前記抗体またはその抗原結合断片。 An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4.
(A) SEQ ID NOs: 6 and 9, respectively;
(B) SEQ ID NOs: 6 and 10, respectively;
(C) SEQ ID NOs: 6 and 11;
(D) SEQ ID NOs: 7 and 9, respectively;
(E) SEQ ID NOs: 7 and 10, respectively;
(F) SEQ ID NOs: 7 and 11, respectively;
(G) SEQ ID NOs: 8 and 9, respectively;
The antibody or antigen-binding fragment thereof comprising (h) a heavy chain variable region and a light chain variable region containing the amino acid sequences of SEQ ID NOs: 8 and 11, respectively; or (i).
(a)それぞれ配列番号16及び19;
(b)それぞれ配列番号16及び30;
(c)それぞれ配列番号16及び31;
(d)それぞれ配列番号17及び19;
(e)それぞれ配列番号17及び30;
(f)それぞれ配列番号17及び31;
(g)それぞれ配列番号18及び19;
(h)それぞれ配列番号18及び30;
(i)それぞれ配列番号18及び31;
(j)それぞれ配列番号89及び19;
(k)それぞれ配列番号90及び19;または
(l)それぞれ配列番号91及び19のアミノ酸配列を含む重鎖及び軽鎖を含む、請求項1~3のいずれか1項に記載の抗体またはその抗原結合断片。 The antibody or antigen-binding fragment
(A) SEQ ID NOs: 16 and 19;
(B) SEQ ID NOs: 16 and 30, respectively;
(C) SEQ ID NOs: 16 and 31, respectively;
(D) SEQ ID NOs: 17 and 19, respectively;
(E) SEQ ID NOs: 17 and 30, respectively;
(F) SEQ ID NOs: 17 and 31, respectively;
(G) SEQ ID NOs: 18 and 19, respectively;
(H) SEQ ID NOs: 18 and 30, respectively;
(I) SEQ ID NOs: 18 and 31, respectively;
(J) SEQ ID NOs: 89 and 19;
(K) The antibody or antigen thereof according to any one of claims 1 to 3 , which comprises a heavy chain and a light chain containing the amino acid sequences of SEQ ID NOs: 90 and 19; or (l), respectively. Bonded fragment.
The therapeutic antibody or antigen binding fragment comprises (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 55, or (ii) of SEQ ID NO: 64. The composition or kit according to any one of claims 20 to 27 , comprising a heavy chain variable region comprising an amino acid sequence and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 65.
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