JP2016020349A5

JP2016020349A5 -

Info

Publication number: JP2016020349A5
Application number: JP2015147748A
Authority: JP
Filing date: 2015-07-27
Publication date: 2016-03-17

Claims

A binding protein comprising first and second polypeptide chains, wherein the first polypeptide chain is VD1- (X1) _n- VD2-C- (X2) _n
(VD1 is a first heavy chain variable domain,
VD2 is a second heavy chain variable domain,
C is a constant domain;
X1 is a linker (where, X1 is not a CH1), and X2, Ru Fc region or a variant Fc region der)
And the second polypeptide chain is VD1- (X1) _n- VD2-C- (X2) _n
(VD1 is a first light chain variable domain,
VD2 is a second light chain variable domain,
C is a constant domain;
X1 is a linker (where X1 is not CL ) and X2 does not contain an Fc region )
Including
Where n is 0 or 1 and the first and second polypeptide chains of the binding protein form two functional antigen binding sites, each antigen binding site binding to TNF or IL-17 can do,
Binding protein.

A binding protein comprising first and second polypeptide chains, wherein the first polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first heavy chain variable domain;
VD2 is the second heavy chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CH1), and
X2 is an Fc region or a variant Fc region)
Including: and
Said second polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first light chain variable domain;
VD2 is the second light chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CL), and
X2 does not include the Fc region)
Including
Where n is 0 or 1 and the first and second polypeptide chains of the binding protein form two functional antigen binding sites, each antigen binding site being an IL-1α or IL-1β Can be combined with the
Binding protein.

A binding protein comprising first and second polypeptide chains, wherein the first polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first heavy chain variable domain;
VD2 is the second heavy chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CH1), and
X2 is an Fc region or a variant Fc region)
Including: and
Said second polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first light chain variable domain;
VD2 is the second light chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CL), and
X2 does not include the Fc region)
Including
Where n is 0 or 1 and the first and second polypeptide chains of the binding protein form two functional antigen binding sites, each antigen binding site being an IL-17 or IL-1β Can be combined with the
Binding protein.

A binding protein comprising first and second polypeptide chains, wherein the first polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first heavy chain variable domain;
VD2 is the second heavy chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CH1), and
X2 is an Fc region or a variant Fc region)
Including: and
Said second polypeptide chain is VD1- (X1) _ｎn -VD2-C- (X2) _ｎn
(VD1 is the first light chain variable domain;
VD2 is the second light chain variable domain;
C is a constant domain;
X1 is a linker (where X1 is not CL), and
X2 does not include the Fc region)
Including
Where n is 0 or 1 and the first and second polypeptide chains of the binding protein form two functional antigen binding sites, each antigen binding site comprising TNF, IL-17, IL Can bind to any one of -1α, IL-1β, VEGF, ErbB3, EGFR and CD,
Binding protein.

The binding protein according to any one of claims 1 to 4, comprising two first polypeptide chains, two second polypeptide chains and four functional antigen binding sites.

Before hear varying domains, variable domains of the murine variable domains of human, in any one of claims 1 to 5 selected from the group consisting of variable domains and humanized Tunable domains CDR-grafted The binding protein described.

X1 _{is, AKTTPKLEEGEFSEAR; AKTTPKLEEGEFSEARV; AKTTPKLGG; RADAAP} ; RADAAPTVS; RADAAAAGGPGS; RADAAAA (G 4 S) 4; SAKTTP; SAKTTPKLGG; SAKTTPKLEEGEFSEARV; ADAAP; ADAAPTVSIFPP; TVAAP; TVAAPSVFIFPP; QPKAAP; QPKAAPSVTLFPP; AKTTPP; AKTTPPSVTPLAP; AKTTAP; AKTTAPSVYPLAP; ASTKGP ; ASTKGPSVFPLAP; GGGGSGGGGSGGGGS; GENKVEYAPALMALS; GPAKELTPLKEAKVS ; and it is selected from the group consisting of GHEAAAVMQVQYPAS Ruri Lanka The binding protein according to any one of claims 1 to 6, wherein

The binding protein according to any one of claims 1 to 7, wherein the binding protein does not contain X2.

The binding protein, one or more binds to the target antigen, or one or can be neutralize more of the target antigen binding protein according to any one of claims 1-8.

The binding protein is at least 10 as measured by surface plasmon resonance. ^２2 M ^−１-1 S ^−１-1 At least 10 ^３3 M ^−１-1 S ^−１-1 At least about 10 ^４4 M ^−１-1 S ^−１-1 At least 10 ^５5 M ^−１-1 S ^−１-1 And at least 10 ^６6 M ^−１-1 S ^−１-1 A binding rate constant (K) to said one or more target antigens selected from the group consisting of _ｏｎon The binding protein according to any one of claims 1 to 9, wherein

The binding protein is up to 10 when measured by surface plasmon resonance. ^−３-3 s ^−１-1 And up to 10 ^−４-4 s ^−１-1 Dissociation rate constant (K) to the one or more targets selected from the group consisting of _ｏｆｆoff The binding protein according to any one of claims 1 to 9, wherein

Up to 10 binding proteins ^−７-7 M; up to 10 ^−８-8 M; up to 10 ^−９-9 M; and up to 10 ^−１０-10 Dissociation constant (K) for the one or more targets selected from the group consisting of M _ＤD The binding protein according to any one of claims 1 to 9, wherein

A binding protein conjugate comprising the binding protein according to any one of claims 1 to 12, and further comprising an agent selected from the group consisting of an immunoadhesion molecule, a contrast agent, a therapeutic agent and a cytotoxic agent.

The binding protein conjugate according to claim 13, wherein the agent is a contrast agent selected from the group consisting of a radioactive label, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

The contrast agent ^３3 H, ^１４14 C, ^３５35 S, ^９０90 Y, ^９９99 Tc, ^１１１111 In, ^１２５125 I, ^１３１131 I, ^１７７177 Lu, ^１６６166 Ho and ^１５３153 15. The binding protein conjugate of claim 14, wherein the binding protein conjugate is a radioactive label selected from the group consisting of Sm.

The agent is a therapeutic agent or cytotoxicity selected from the group consisting of metabolic inhibitors, alkylating agents, antibiotics, growth factors, cytokines, anti-angiogenic agents, anti-mitotic agents, anthracyclines, toxins and apoptotic agents. The binding protein conjugate according to claim 13, which is an agent.

The binding protein according to any one of claims 1 to 12, wherein the binding protein is a crystallized binding protein.

18. A crystallized binding protein according to claim 17, wherein the binding protein has a longer half-life in vivo than the soluble counterpart of the binding protein.

The crystallized binding protein of claim 17, wherein the binding protein retains biological activity.

An isolated nucleic acid encoding the amino acid sequence of the binding protein of any one of claims 1-12.

21. A vector comprising the isolated nucleic acid of claim 20.

The vector according to claim 21, wherein the vector is selected from the group consisting of pcDNA (TM), pTT, pTT3, pEFBOS, pBV, pJV, pcDNA (TM) 3.1 TOPO (R), pEF6TOPO (R) and pBJ. The described vector.

A host cell comprising the vector according to claim 21 or 22.

24. The host cell of claim 23, wherein the host cell is a prokaryotic cell or a eukaryotic cell.

The prokaryotic cell is E. coli. 25. A host cell according to claim 24 which is E. coli.

25. A host cell according to claim 24, wherein the eukaryotic cell is selected from the group consisting of protist cells, animal cells, plant cells and fungal cells.

27. The host cell according to claim 26, wherein the animal cell is selected from the group consisting of a mammalian cell, an avian cell and an insect cell.

27. The host cell according to claim 26, selected from the group consisting of CHO cells, COS cells and Sf9 cells.

27. The host cell of claim 26, wherein the fungal cell is a yeast cell.

30. A method for producing a binding protein comprising culturing the host cell of any one of claims 23-29 in a medium under conditions sufficient to produce the binding protein.

31. The method of claim 30, wherein 50% to 75% of the produced binding protein is the bispecific binding protein of any one of claims 1-12.

31. The method of claim 30, wherein 75% to 90% of the produced binding protein is a bispecific binding protein according to any one of claims 1-12.

31. The method of claim 30, wherein 90% to 95% of the produced binding protein is the bispecific binding protein of any one of claims 1-12.

31. A protein produced according to the method of claim 30.

A pharmaceutical composition comprising the binding protein according to any one of claims 1 to 12 and 34 and a pharmaceutically acceptable carrier.

36. The pharmaceutical composition of claim 35, further comprising at least one additional therapeutic agent.

Said additional agent is cytotoxic agent, angiogenesis inhibitor; kinase inhibitor; costimulatory molecule blocker; adhesion molecule blocker; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporine; rapamycin; FK506; Label or reporter; TNF antagonist; anti-rheumatic drug; muscle relaxant, anesthetic, non-steroidal anti-inflammatory drug (NSAID), analgesic, anesthetic, sedative, local anesthesia, neuromuscular blocking agent, antimicrobial agent, anti-psoriasis Agent, corticosteroid, anabolic steroid, erythropoietin, immunization, immunoglobulin, immunosuppressant, growth hormone, hormone replacement agent, radiopharmaceutical, antidepressant, antipsychotic, stimulant, asthma, beta agonist, for inhalation Steroids, epinephrine or analogs, cytokines and cytokine antagonists Selected from the group consisting of the pharmaceutical composition according to claim 36.

Parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intrabronchial, intraperitoneal, intracapsular, intrachondral, intracavity, intracavity, intracerebellum, intraventricular, intracolonic, intracervical, stomach Intra, intrahepatic, intramyocardial, intraosseous, intrapelvic, pericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intracolonial, intrarenal, intraretinal, intrathecal, intrasynovial, intrathoracic, uterus 38. The pharmaceutical composition according to claims 35-37, suitable for administration to a subject in at least one manner selected from internal, intravesical, bolus, vaginal, rectal, buccal, sublingual, intranasal and transdermal.

35. A binding protein according to any one of claims 1 to 12 and 34 for use in the treatment of a disease or disorder of interest.

35. Use of a binding protein according to any one of claims 1 to 12 and 34 in the manufacture of a medicament for the treatment of a disease or disorder.

The binding protein is parenteral, subcutaneous, intramuscular, intravenous, intraarticular, intrabronchial, intraperitoneal, intracapsular, intrachondral, intracavity, intracerebral, intracerebellum, intraventricular, intracolonic, Intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intraperitoneal, intraperitoneal, intrapleural, prostate, intrapulmonary, intracolonial, intrarenal, intraretinal, intraspinal, synovial, 41. Formulated for administration by at least one mode selected from intrathoracic, intrauterine, intravesical, bolus, vaginal, rectal, buccal, sublingual, intranasal and transdermal. use.

The disease or disorder is rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthritis, systemic lupus erythematosis, Crohn's disease, ulcerative colitis, inflammation The use according to claim 40 or 41, which is sexual bowel disease, colon cancer, rectal cancer, ovarian cancer, breast cancer, lung cancer, non-small cell lung cancer, or pancreatic cancer.