JPWO2019126401A5 - - Google Patents
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- JPWO2019126401A5 JPWO2019126401A5 JP2020533728A JP2020533728A JPWO2019126401A5 JP WO2019126401 A5 JPWO2019126401 A5 JP WO2019126401A5 JP 2020533728 A JP2020533728 A JP 2020533728A JP 2020533728 A JP2020533728 A JP 2020533728A JP WO2019126401 A5 JPWO2019126401 A5 JP WO2019126401A5
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関連する実施形態において、本開示は、Wntシグナリングの増加または強化に関連する疾患または障害を有する対象を治療するための方法であって、対象に、本明細書で開示されている、Wntシグナリング経路の阻害物質である単離抗体またはその抗原結合フラグメントを含む医薬組成物の有効量を投与することを含む、方法を提供する。ある特定の実施形態において、疾患または障害は、腫瘍及び癌、変性性障害、線維症、心不全、冠動脈疾患、異所性骨化、骨粗鬆症、及び先天性高骨量障害からなる群より選択される。
特定の実施形態では、例えば、以下が提供される:
(項目1)
1つ以上のLRP5またはLRP6受容体に結合する単離抗体またはその抗原結合フラグメントであって、下記を含む配列:
(i)表1Aのいずれかの抗体について記載されたCDRH1、CDRH2、及びCDRH3配列、
(ii)表1Aのいずれかの抗体について記載されたCDRL1、CDRL2、及びCDRL3配列、ならびに/または
(iii)表2のいずれかの抗体について記載されたCDRH1、CDRH2、CDRH3、CDRL1、CDRL2、及びCDRL3配列、を含む、単離抗体またはその抗原結合フラグメント、
あるいは、1つ以上のアミノ酸修飾を含む前記抗体のバリアントまたはその抗原結合フラグメントであって、前記CDR配列中に8つ未満のアミノ酸置換を含む、前記抗体のバリアントまたはその抗原結合フラグメント。
(項目2)
配列番号1~24のいずれかに記載されたアミノ酸配列に対し少なくとも90%の同一性を有するアミノ酸配列を含む重鎖可変領域を含む、項目1に記載の単離抗体またはその抗原結合フラグメント。
(項目3)
配列番号1~24のいずれかに記載されたアミノ酸配列を含む重鎖可変領域を含む、項目2に記載の単離抗体またはその抗原結合フラグメント。
(項目4)
前記抗体またはその抗原結合フラグメントがヒト化されている、項目1~3のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目5)
前記抗体またはその抗原結合フラグメントが、1本鎖抗体、scFv、ヒンジ領域を欠いた1価抗体、VHHもしくはsdAb、またはミニボディである、項目1~5のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目6)
前記抗体またはその抗原結合フラグメントがVHH VHHまたはsdAbである、項目5に記載の単離抗体またはその抗原結合フラグメント。
(項目7)
前記抗体またはその抗原結合フラグメントがFabまたはFab’フラグメントである、項目1に記載の単離抗体またはその抗原結合フラグメント。
(項目8)
前記抗体またはその抗原結合フラグメントが融合タンパク質である、項目1~7のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目9)
前記抗体またはその抗原結合フラグメントが、1つ以上のフリズルド(Fzd)受容体に結合するポリペプチド配列と融合している、項目8に記載の単離抗体またはその抗原結合フラグメント。
(項目10)
1つ以上のFzd受容体に結合する前記ポリペプチド配列が、1つ以上のFzd受容体に結合する抗体またはその抗原結合フラグメントである、項目9に記載の単離抗体またはその抗原結合フラグメント。
(項目11)
前記抗体またはその抗原結合フラグメントがLRP5に結合する、項目1~10のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目12)
前記抗体またはその抗原結合フラグメントがLRP6に結合する、項目11に記載の単離抗体またはその抗原結合フラグメント。
(項目13)
前記抗体またはその抗原結合フラグメントがLRP5及びLRP6に結合する、項目12に記載の単離抗体またはその抗原結合フラグメント。
(項目14)
ヒトLRP5またはLRP6との結合において項目1~13のいずれかに記載の抗体と競合する、単離抗体またはその抗原結合フラグメント。
(項目15)
前記LRP5またはLRP6に50μM以下のKDで結合する、項目1~14のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目16)
細胞内の、任意選択で哺乳類細胞内のWntシグナリング経路を調節する、項目1~15のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目17)
前記細胞内の前記Wntシグナリング経路を介してシグナリングを増加させる、項目16に記載の単離抗体またはその抗原結合フラグメント。
(項目18)
前記細胞内の前記Wntシグナリング経路を介してシグナリングを減少させる、項目16に記載の単離抗体またはその抗原結合フラグメント。
(項目19)
前記Wntシグナリング経路がカノニカルWntシグナリング経路である、項目16~18のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目20)
前記Wntシグナリング経路が非カノニカルWntシグナリング経路である、項目16~18のいずれかに記載の単離抗体またはその抗原結合フラグメント。
(項目21)
項目1~20のいずれかに記載の単離抗体またはその抗原結合フラグメントをコードする、単離ポリヌクレオチド。
(項目22)
項目22に記載の単離ポリヌクレオチドを含む、発現ベクター。
(項目23)
項目22に記載の発現ベクターを含む、単離宿主細胞。
(項目24)
生理的に許容される賦形剤、希釈剤、または担体と、項目1~20または33のいずれかに記載の単離抗体またはその抗原結合フラグメントの治療有効量とを含む、医薬組成物。
(項目25)
細胞内のWntシグナリング経路を刺激するための方法であって、前記細胞を、項目17に記載の単離抗体またはその抗原結合フラグメントに接触させることを含む、方法。
(項目26)
前記抗体またはその抗原結合フラグメントが、1つ以上のフリズルド(Fzd)受容体に結合するポリペプチド配列を含む融合タンパク質である、項目24に記載の方法。
(項目27)
細胞内のWntシグナリング経路を阻害するための方法であって、前記細胞を、項目18に記載の単離抗体またはその抗原結合フラグメントに接触させることを含む、方法。
(項目28)
Wntシグナリング低減に関連する疾患または障害を有する対象を治療するための方法であって、前記対象に、項目24に記載の医薬組成物の有効量を投与することを含み、前記単離抗体またはその抗原結合フラグメントがWntシグナリング経路のアゴニストである、方法。
(項目29)
前記疾患または障害が、骨折、疲労骨折、脊椎圧迫骨折、変形性関節症、骨粗鬆症、骨粗鬆症性骨折、癒合不全骨折、癒合遅延骨折、脊椎固定、脊椎手術のための術前最適化、骨壊死、移植または整形外科デバイスの骨結合、骨形成不全症、骨移植、腱修復、腱-骨結合、歯の成長及び再生、顎顔面手術、歯科インプラント、歯周病、顎顔面再構築、顎、臀部、または大腿骨頭の骨壊死、無血管性壊死、脱毛症、難聴、前庭機能低下、黄斑変性、加齢性黄斑変性(AMD)、硝子体網膜症、網膜症、糖尿病性網膜症、網膜変性疾患、フックスジストロフィー、角膜疾患、卒中、外傷性脳損傷、アルツハイマー病、多発性硬化症、筋ジストロフィー、サルコペニア及びカヘキシーにおける筋萎縮、血液脳関門(BBB)に影響を及ぼす疾患、脊髄損傷、脊髄疾患、口腔粘膜炎、短腸症候群、炎症性腸疾患(IBD)、メタボリックシンドローム、糖尿病、脂質異常症、膵炎、膵外分泌機能不全、創傷治癒、糖尿病性足部潰瘍、褥瘡、静脈性下腿潰瘍、表皮水疱症、皮膚形成不全、心筋梗塞、冠動脈疾患、心不全、造血細胞障害、免疫不全、移植片対宿主病、急性腎損傷、慢性腎疾患、慢性閉塞性肺疾患(COPD)、特発性肺線維症、あらゆる原因の急性肝不全、薬物誘発性急性肝不全、アルコール性肝疾患、あらゆる原因の慢性肝不全、肝硬変、あらゆる原因の肝線維症、門脈圧亢進症、あらゆる原因の慢性肝不全、非アルコール性脂肪性肝炎(NASH)、非アルコール性脂肪性肝疾患(NAFLD)(脂肪肝)、アルコール性肝炎、C型肝炎ウイルス誘発性肝疾患(HCV)、B型肝炎ウイルス誘発性肝疾患(HBV)、その他のウイルス性肝炎(例えば、A型肝炎ウイルス誘発性肝疾患(HAV)及びD型肝炎ウイルス誘発性肝疾患(HDV))、原発性胆汁性肝硬変、自己免疫性肝炎、肝臓手術、肝損傷、肝移植、肝臓の手術及び移植における「過小グラフト(small for size)」症候群、先天性肝疾患及び肝障害、遺伝性疾患、変性、加齢、薬物、または損傷に起因するその他の任意の肝障害または検出からなる群より選択される、項目28に記載の方法。
(項目30)
Wntシグナリング増加または強化に関連する疾患または障害を有する対象を治療するための方法であって、前記対象に、項目24に記載の医薬組成物の有効量を投与することを含み、前記単離抗体またはその抗原結合フラグメントがWntシグナリング経路の阻害物質である、方法。
(項目31)
前記疾患または障害が、腫瘍及び癌、変性性障害、任意の臓器または組織の線維症、心不全、冠動脈疾患、異所性骨化、骨粗鬆症、及び先天性高骨量障害からなる群より選択される、項目29に記載の方法。
(項目32)
LRP6受容体に結合する単離抗体またはその抗原結合フラグメントであって、前記LRP6受容体のE3 β-プロペラ領域、またはLRP5受容体の対応領域に結合する、単離抗体またはその抗原結合フラグメント。
(項目33)
LRP6受容体に結合する単離抗体またはその抗原結合フラグメントであって、アミノ酸残基637~878を含むまたはそれからなる前記LRP6受容体の領域内の1つ以上のエピトープ、またはLRP5受容体の対応領域に結合する、単離抗体またはその抗原結合フラグメント。
(項目34)
1つ以上のLRP受容体に結合する単離抗体またはその抗原結合フラグメントであって、表3に示されるいずれかのアミノ酸残基のセットにて、5オングストローム未満の距離で前記LRP受容体に接触する、単離抗体またはその抗原結合フラグメント。
In a related embodiment, the present disclosure is a method for treating a subject having a disease or disorder associated with an increase or enhancement of Wnt signaling, the Wnt signaling pathway disclosed herein to the subject. Provided is a method comprising administering an effective amount of a pharmaceutical composition comprising an isolated antibody or an antigen-binding fragment thereof which is an inhibitor of the above. In certain embodiments, the disease or disorder is selected from the group consisting of tumors and cancers, degenerative disorders, fibrosis, heart failure, coronary artery disease, ectopic ossification, osteoporosis, and congenital high bone mass disorders. ..
In certain embodiments, for example, the following are provided:
(Item 1)
An isolated antibody or antigen-binding fragment thereof that binds to one or more LRP5 or LRP6 receptors, comprising the following sequences:
(I) CDRH1, CDRH2, and CDRH3 sequences described for any of the antibodies in Table 1A.
(Ii) The CDRL1, CDRL2, and CDRL3 sequences described for any of the antibodies in Table 1A, and / or
(Iii) An isolated antibody or antigen-binding fragment thereof comprising the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences described for any of the antibodies in Table 2.
Alternatively, a variant of the antibody or an antigen-binding fragment thereof, the variant of the antibody or an antigen-binding fragment thereof, comprising one or more amino acid modifications and comprising less than eight amino acid substitutions in the CDR sequence.
(Item 2)
The isolated antibody or antigen-binding fragment thereof according to item 1, which comprises a heavy chain variable region containing an amino acid sequence having at least 90% identity with respect to the amino acid sequence set forth in any of SEQ ID NOs: 1 to 24.
(Item 3)
The isolated antibody or antigen-binding fragment thereof according to item 2, which comprises a heavy chain variable region comprising the amino acid sequence set forth in any of SEQ ID NOs: 1 to 24.
(Item 4)
The isolated antibody or antigen-binding fragment thereof according to any one of Items 1 to 3, wherein the antibody or antigen-binding fragment thereof is humanized.
(Item 5)
Item 6. The isolated antibody according to any one of Items 1 to 5, wherein the antibody or an antigen-binding fragment thereof is a single-chain antibody, scFv, a monovalent antibody lacking a hinge region, VHH or sdAb, or a minibody. Antigen binding fragment.
(Item 6)
Item 5. The isolated antibody or antigen-binding fragment thereof according to item 5, wherein the antibody or antigen-binding fragment thereof is VHH VHH or sdAb.
(Item 7)
The isolated antibody or antigen-binding fragment thereof according to item 1, wherein the antibody or antigen-binding fragment thereof is a Fab or Fab'fragment.
(Item 8)
The isolated antibody or antigen-binding fragment thereof according to any one of Items 1 to 7, wherein the antibody or antigen-binding fragment thereof is a fusion protein.
(Item 9)
Item 8. The isolated antibody or antigen-binding fragment thereof according to item 8, wherein the antibody or antigen-binding fragment thereof is fused with a polypeptide sequence that binds to one or more frizzled (Fzd) receptors.
(Item 10)
Item 9. The isolated antibody or antigen-binding fragment thereof according to item 9, wherein the polypeptide sequence that binds to one or more Fzd receptors is an antibody that binds to one or more Fzd receptors or an antigen-binding fragment thereof.
(Item 11)
The isolated antibody or antigen-binding fragment thereof according to any one of Items 1 to 10, wherein the antibody or an antigen-binding fragment thereof binds to LRP5.
(Item 12)
The isolated antibody or antigen-binding fragment thereof according to item 11, wherein the antibody or an antigen-binding fragment thereof binds to LRP6.
(Item 13)
Item 12. The isolated antibody or antigen-binding fragment thereof according to item 12, wherein the antibody or antigen-binding fragment thereof binds to LRP5 and LRP6.
(Item 14)
An isolated antibody or antigen-binding fragment thereof that competes with the antibody according to any one of items 1 to 13 in binding to human LRP5 or LRP6.
(Item 15)
The isolated antibody or antigen-binding fragment thereof according to any one of Items 1 to 14, which binds to LRP5 or LRP6 with a KD of 50 μM or less.
(Item 16)
The isolated antibody or antigen-binding fragment thereof according to any one of items 1 to 15, which regulates the Wnt signaling pathway in the intracellular, optionally in mammalian cells.
(Item 17)
Item 16. The isolated antibody or antigen-binding fragment thereof according to item 16, which increases signaling through the Wnt signaling pathway in the cell.
(Item 18)
16. The isolated antibody or antigen-binding fragment thereof according to item 16, which reduces signaling through the Wnt signaling pathway in the cell.
(Item 19)
The isolated antibody or antigen-binding fragment thereof according to any one of items 16 to 18, wherein the Wnt signaling pathway is a canonical Wnt signaling pathway.
(Item 20)
The isolated antibody or antigen-binding fragment thereof according to any one of items 16 to 18, wherein the Wnt signaling pathway is a non-canonical Wnt signaling pathway.
(Item 21)
An isolated polynucleotide encoding the isolated antibody according to any one of items 1 to 20 or an antigen-binding fragment thereof.
(Item 22)
An expression vector comprising the isolated polynucleotide according to item 22.
(Item 23)
An isolated host cell comprising the expression vector according to item 22.
(Item 24)
A pharmaceutical composition comprising a physiologically acceptable excipient, diluent, or carrier and a therapeutically effective amount of the isolated antibody or antigen-binding fragment thereof according to any one of items 1-20 or 33.
(Item 25)
A method for stimulating an intracellular Wnt signaling pathway, comprising contacting the cell with an isolated antibody or antigen-binding fragment thereof according to item 17.
(Item 26)
24. The method of item 24, wherein the antibody or antigen binding fragment thereof is a fusion protein comprising a polypeptide sequence that binds to one or more Frizzled (Fzd) receptors.
(Item 27)
A method for inhibiting the intracellular Wnt signaling pathway, comprising contacting the cell with an isolated antibody or antigen-binding fragment thereof according to item 18.
(Item 28)
A method for treating a subject having a disease or disorder associated with Wnt signaling reduction, comprising administering to the subject an effective amount of the pharmaceutical composition according to item 24, the isolated antibody or the isolated antibody thereof. A method in which the antigen binding fragment is an agonist of the Wnt signaling pathway.
(Item 29)
The diseases or disorders include fractures, fatigue fractures, spinal compression fractures, osteoarthritis, osteoporosis, osteoporotic fractures, incomplete fusion fractures, delayed fusion fractures, spinal fusion, preoperative optimization for spinal surgery, osteonecrosis, Bone connection of implantable or orthopedic devices, osteodysplasia, bone transplantation, tendon repair, tendon-bone connection, tooth growth and regeneration, maxillofacial surgery, dental implants, periodontal disease, maxillofacial reconstruction, jaw, buttocks , Or femoral head osteonecrosis, avascular necrosis, alopecia, hearing loss, vestibular dysfunction, yellow spot degeneration, age-related yellow spot degeneration (AMD), vitreous retinopathy, retinopathy, diabetic retinopathy, retinal degenerative disease , Fuchs dystrophy, corneal disease, stroke, traumatic brain injury, Alzheimer's disease, multiple sclerosis, muscular dystrophy, muscle atrophy in sarcopenia and cahexy, diseases affecting the blood-brain barrier (BBB), spinal cord injury, spinal cord disease, oral cavity Mucositis, short bowel syndrome, inflammatory bowel disease (IBD), metabolic syndrome, diabetes, dyslipidemia, pancreatitis, pancreatic exocrine dysfunction, wound healing, diabetic foot ulcer, decubitus, venous lower leg ulcer, epidermal vesicular disease , Skin dysplasia, myocardial infarction, coronary artery disease, heart failure, hematopoietic cell disorder, immunodeficiency, transplant-to-host disease, acute renal injury, chronic renal disease, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, all Causes of acute liver failure, drug-induced acute liver failure, alcoholic liver disease, chronic liver failure of all causes, liver cirrhosis, liver fibrosis of all causes, hypertonic hypertension, chronic liver failure of all causes, non-alcoholic Fatty hepatitis (NASH), non-alcoholic fatty liver disease (NAFLD) (lipohepatitis), alcoholic hepatitis, hepatitis C virus-induced liver disease (HCV), hepatitis B virus-induced liver disease (HBV), Other viral hepatitis (eg, hepatitis A virus-induced liver disease (HAV) and hepatitis D virus-induced liver disease (HDV)), primary biliary cirrhosis, autoimmune hepatitis, liver surgery, liver injury, "Small for size" syndrome in liver transplantation, liver surgery and transplantation, congenital liver disease and liver damage, hereditary disease, degeneration, aging, drugs, or any other liver damage due to injury 28. The method of item 28, selected from the group consisting of detections.
(Item 30)
A method for treating a subject having a disease or disorder associated with increased or enhanced Wnt signaling, comprising administering to the subject an effective amount of the pharmaceutical composition according to item 24, said isolated antibody. Or a method in which the antigen binding fragment is an inhibitor of the Wnt signaling pathway.
(Item 31)
The disease or disorder is selected from the group consisting of tumors and cancers, degenerative disorders, fibrosis of any organ or tissue, heart failure, coronary artery disease, ectopic ossification, osteoporosis, and congenital high bone mass disorders. , Item 29.
(Item 32)
An isolated antibody or antigen-binding fragment thereof that binds to the LRP6 receptor and that binds to the E3 β-propeller region of the LRP6 receptor or the corresponding region of the LRP5 receptor.
(Item 33)
An isolated antibody that binds to the LRP6 receptor or an antigen-binding fragment thereof, the one or more epitopes within the region of the LRP6 receptor comprising or consisting of amino acid residues 637-878, or the corresponding region of the LRP5 receptor. An isolated antibody or antigen-binding fragment thereof that binds to.
(Item 34)
An isolated antibody that binds to one or more LRP receptors or an antigen-binding fragment thereof that contacts the LRP receptor at a distance of less than 5 angstroms with any set of amino acid residues shown in Table 3. An isolated antibody or antigen-binding fragment thereof.
Claims (36)
(i)表1Aに記載された配列番号107のCDRH1配列、配列番号203のCDRH2配列、及び配列番号452のCDRH3配列、 (I) the CDRH1 sequence of SEQ ID NO: 107, the CDRH2 sequence of SEQ ID NO: 203, and the CDRH3 sequence of SEQ ID NO: 452, which are shown in Table 1A.
を含む、単離抗体またはその抗原結合フラグメント、An isolated antibody or antigen-binding fragment thereof, including
あるいは、1つ以上のアミノ酸修飾を含む前記抗体のバリアントまたはその抗原結合フラグメントであって、前記CDR配列中に8つ未満のアミノ酸置換を含む、前記抗体のバリアントまたはその抗原結合フラグメント。 Alternatively, a variant of the antibody or an antigen-binding fragment thereof, the variant of the antibody or an antigen-binding fragment thereof, comprising one or more amino acid modifications and comprising less than eight amino acid substitutions in the CDR sequence.
(i)表1Aのいずれかの抗体について記載されたCDRH1、CDRH2、及びCDRH3配列、
(ii)表1Aのいずれかの抗体について記載されたCDRL1、CDRL2、及びCDRL3配列、ならびに/または
(iii)表2のいずれかの抗体について記載されたCDRH1、CDRH2、CDRH3、CDRL1、CDRL2、及びCDRL3配列、を含む、単離抗体またはその抗原結合フラグメント、
あるいは、1つ以上のアミノ酸修飾を含む前記抗体のバリアントまたはその抗原結合フラグメントであって、前記CDR配列中に8つ未満のアミノ酸置換を含む、前記抗体のバリアントまたはその抗原結合フラグメント。 An isolated antibody or antigen-binding fragment thereof that binds to one or more LRP5 or LRP6 receptors, comprising the following sequences:
(I) CDRH1, CDRH2, and CDRH3 sequences described for any of the antibodies in Table 1A.
(Ii) the CDRL1, CDRL2, and CDRL3 sequences described for any antibody of Table 1A and / or (iii) the CDRH1, CDRH2, CDRH3, CDRL1, and / or (iii) described for any antibody of Table 2; An isolated antibody or antigen-binding fragment thereof, comprising a CDRL3 sequence,
Alternatively, a variant of the antibody or an antigen-binding fragment thereof, the variant of the antibody or an antigen-binding fragment thereof, comprising one or more amino acid modifications and comprising less than eight amino acid substitutions in the CDR sequence.
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