WO2018021450A1 - 増強されたfviii補因子機能代替活性を有する二重特異性抗体 - Google Patents
増強されたfviii補因子機能代替活性を有する二重特異性抗体 Download PDFInfo
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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Definitions
- the present invention relates to an antibody having enhanced FVIII cofactor function substitution activity, a pharmaceutical preparation containing such an antibody as an active ingredient, a method for producing the antibody, and the like. More specifically, the present invention relates to a bispecific antibody having higher FVIII cofactor function substitution activity than ACE910 (Emicizumab).
- Hemophilia A is a bleeding disorder caused by a reduced function or deficiency of congenital blood coagulation factor VIII (FVIII).
- FVIII preparations are usually administered (on-demand administration).
- FVIII preparations are administered prophylactically to prevent bleeding events (Non-patent Documents 1 and 2) (preventive administration).
- the blood half-life of the FVIII preparation is about 12 to 16 hours. Therefore, for continuous prevention, FVIII preparations are administered to patients 3 times a week (Non-patent Documents 3 and 4).
- FVIII preparations are additionally administered at regular intervals as necessary.
- administration of the FVIII preparation is performed intravenously. Therefore, there has been a strong demand for a drug that has a lower administration burden than the FVIII preparation.
- Inhibitors against FVIII occur in hemophilia patients. Inhibitors counteract the effects of the FVIII formulation.
- a bypass formulation is administered for bleeding in patients with inhibitors (inhibitor patients). Their mechanism of action is independent of the function of FVIII, ie the ability to catalyze the activation of blood coagulation factor X (FX) by activated blood coagulation factor IX (FIXa). Therefore, there are cases where the bypass preparation cannot sufficiently stop bleeding. Therefore, there has been a strong demand for drugs that are independent of the presence of inhibitors and that substitute for the function of FVIII.
- Bispecific antibodies against FIXa and FX can substitute for FVIII function by demonstrating FVIII cofactor function substitution activity by positioning both factors in the vicinity (Non-patent Document 5).
- the FVIII cofactor function alternative activity of bispecific antibody against FIXa and FX was calculated from FX activation reaction test with FIXa using colorimetric assay and thrombin generation test using hemophilia A plasma Refers to activity. It has been reported that the FVIII cofactor function alternative activity of the antibody can be improved by optimizing the affinity for FIXa and FX (Non-patent Document 6).
- Non-patent Document 7 ACE910 (Emicizumab), which is one of the antibodies and has high FVIII cofactor function substitution activity, has been reported to exert a hemostatic effect in a monkey hemophilia model (Non-patent Documents 8 and 9). Furthermore, ACE910 (Emicizumab) has been confirmed to have excellent pharmacokinetics (long half-life) and tolerability in clinical trials for healthy individuals (Non-patent Document 10). In clinical trials for patients, ACE910 (Emicizumab) administration significantly reduced the number of bleedings compared to before administration of ACE910 (Emicizumab) (Non-patent Document 11).
- ACE910 (Emicizumab) was found to have an effect of suppressing bleeding frequency in clinical trials, but it was caused by ACE910 (Emicizumab) in the maximum thrombin generation amount (Peak height) in the in vitro thrombin generation test using FVIII-deficient plasma.
- the improvement effect is low compared with the normal level of 100 U / dL FVIII activity (Non-patent Document 8), so that further enhancement of drug efficacy is desired, and further reduction of dosage due to improved specific activity is possible Bispecific antibodies are needed.
- ACE910 introduces a number of amino acid substitutions into the lead antibody using hBS1 obtained by humanizing anti-FIX antibody and / or anti-FIXa antibody and anti-FX antibody obtained from animal immunization as the lead antibody. It is a bispecific antibody that is multifacetedly optimized and has a high FVIII cofactor function substitution activity (Non-patent Document 6, Patent Document 4). FVIII function replacement that has higher maximum activity than ACE910 (Emicizumab) (maximum activity of FVIII cofactor function replacement activity) and can exhibit FVIII cofactor function replacement activity at a lower concentration than ACE910 (Emicizumab) Bispecific antibodies are required. However, no bispecific antibody having an FVIII cofactor function replacement activity higher than that of ACE910 (Emicizumab) has been reported so far in terms of maximum activity and concentration.
- the present invention has been made in view of the above circumstances, and provides an antibody with enhanced FVIII cofactor function replacement activity, a pharmaceutical preparation containing such an antibody as an active ingredient, a method for producing the same, and the like.
- the purpose is to do. More specifically, mutations in variable region sites of heavy and light chains to produce bispecific antibodies with higher FVIII cofactor function alternative activity than ACE910 (Emicizumab) or novel different from ACE910 (Emicizumab) A light chain CDR sequence, and a bispecific antibody recognizing FIX and / or FIXa and FX having the same mutation or CDR sequence, a method for producing the antibody, a pharmaceutical preparation comprising the antibody as an active ingredient, or It is an object of the present invention to provide a method for treating hemophilia A using the pharmaceutical preparation.
- mutants in which amino acid substitutions have been introduced at various sites in the light chain variable region of ACE910 (Emicizumab). I succeeded in finding it. Also, succeeded in obtaining a new light chain having a sequence different from ACE910 having FVIII cofactor function substitution activity from a human antibody library, and finding an amino acid substitution that enhances FVIII cofactor function substitution activity in the light chain succeeded in.
- mutants were introduced by introducing amino acid substitutions at various sites in the heavy chain variable region of the bispecific antibody using the light chain, amino acid substitutions that enhance FVIII cofactor function replacement activity were successfully found. did.
- a polypeptide comprising an antibody light chain variable domain wherein the antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9 or SEQ ID NO: 47
- An antibody light chain variable domain having the described amino acid sequence wherein K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, numbered according to the Kabat numbering system , A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97, one or more amino acids are any amino acids other than cysteine
- a bispecific antibody that recognizes FIX and / or FIXa and FX, and has the light chain CDR1, 2, 3 amino acid sequence of SEQ ID
- One or more amino acids selected from the group consisting of L33, A34, Q50, A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97, An antibody substituted with any amino acid other than cysteine.
- a bispecific antibody that recognizes FIX and / or FIXa and FX wherein the first polypeptide and the third polypeptide form a pair, and the second polypeptide and the fourth polypeptide Form a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 45, and the second polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 46.
- Each of the third polypeptide and the fourth polypeptide includes the amino acid sequence of the antibody light chain variable domain set forth in SEQ ID NO: 47, and the other polypeptide includes SEQ ID NO: 7 An antibody light chain variable domain having the amino acid sequence of the light chain CDR1, 2, 3 according to 8, 9 or 9, or an antibody light chain variable domain having the amino acid sequence of SEQ ID NO: 47, wherein The other polypeptide is numbered according to the Kabat numbering system, K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, A51, S52, R53, K54, E55, An antibody in which one or more amino acids selected from the group consisting of S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 are substituted with any amino acid other than cysteine.
- a polypeptide comprising an antibody heavy chain variable domain wherein the antibody heavy chain variable domain having the amino acid sequence of heavy chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 1, 2, and 3 or SEQ ID NO: 45
- Antibody heavy chain variable domains having the described amino acid sequences, wherein Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a, S53, G54, Q55, numbered according to the Kabat numbering system , S56, T57, Y58, Y59, R60, R61, E62, V63, K64, G65, R95, T96, G97, R98, E99, Y100, G100a, G100b, G100c, W100d, Y100e, F100f, D101 and Y102
- a polypeptide, wherein one or more amino acids selected from the group are substituted with any amino acid other than cysteine.
- An antibody heavy chain variable domain that recognizes FIX and / or FIXa and FX, and has the heavy chain CDR1, 2, 3 amino acid sequence of SEQ ID NOs: 1, 2, 3 Alternatively, comprising an antibody heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 45, wherein Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a are numbered according to the Kabat numbering system.
- a polypeptide comprising an antibody heavy chain variable domain, wherein the antibody heavy chain variable domain having the amino acid sequence of heavy chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 4, 5, and 6 or SEQ ID NO: 46
- Antibody heavy chain variable domains having the described amino acid sequences, wherein D31, N32, N33, M34, D35, D50, I51, N52, T52a, R53, S54, G55, numbered according to the Kabat numbering system G56, S57, I58, Y59, N60, E61, E62, F63, Q64, D65, R95, K96, S97, Y98, G99, Y100, Y100a, L100b, D101, and E102.
- an antibody heavy chain variable domain that recognizes FIX and / or FIXa and FX and has the heavy chain CDR1, 2, 3 amino acid sequence of SEQ ID NOs: 4, 5, 6 Alternatively, comprising an antibody heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 46, wherein D31, N32, N33, M34, D35, D50, I51, N52, T52a, numbered according to the Kabat numbering system , R53, S54, G55, G56, S57, I58, Y59, N60, E61, E62, F63, Q64, D65, R95, K96, S97, Y98, G99, Y100, Y100a, L100b, D101 and E102 An antibody in which one or more selected amino acids are substituted with any amino acid other than cysteine.
- a bispecific antibody that recognizes FIX and / or FIXa and FX wherein the first polypeptide and the third polypeptide form a pair, and the second polypeptide and the fourth polypeptide Form a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain set forth in SEQ ID NO: 45, the third polypeptide is the amino acid sequence of the antibody light chain set forth in SEQ ID NO: 43, and the fourth polypeptide
- the peptide comprises the amino acid sequence of the antibody light chain set forth in SEQ ID NO: 44, respectively, and the second polypeptide has the amino acid sequence of heavy chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 4, 5, and 6.
- An antibody light chain comprising any one of the following amino acid sequences selected from (a1) to (a6), (b1) to (b23) and (c1) to (c3): (a1) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 13 (QNK131) (a2) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 14 (QNK284) (a3) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 15 (QNK315) (a4) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 16 (QNL182) (a5) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 17 (QNL492) (a6) Antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 18 (QNL576) (b1) an antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 19 (JNK131) (b2) an antibody light chain comprising the amino acid sequence set forth in SEQ ID NO:
- the first polypeptide is (d1) or (d2).
- (D1) An antibody heavy chain variable domain having the amino acid sequence of heavy chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 1, 2, 3 or an antibody heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 45
- Polypeptide (d2) containing antibody heavy chain variable domain having the amino acid sequence of heavy chain CDR1, 2, 3 as set forth in SEQ ID NO: 1, 2, 3 or antibody heavy chain variable having the amino acid sequence as set forth in SEQ ID NO: 45 A polypeptide comprising a domain, numbered according to the Kabat numbering system, Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a, S53, G54, Q55, S56, T57, Y58, Y59 , R60, R61, E62, V63, K64, G65, R95, T96, G97, R98, E99, Y100, G100a, G100b, G100c
- the second polypeptide is (e1) or (e2).
- (E1) comprising an antibody heavy chain variable domain having the amino acid sequence of heavy chain CDR1, 2, 3 as set forth in SEQ ID NOs: 4, 5, 6 or an antibody heavy chain variable domain having the amino acid sequence as set forth in SEQ ID NO: 46
- Polypeptide (e2) antibody heavy chain variable domain having the amino acid sequence of heavy chain CDR1, 2, 3 described in SEQ ID NO: 4, 5, 6 or antibody heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 46
- a polypeptide comprising: D31, N32, N33, M34, D35, D50, I51, T52a, N52, R53, S54, G55, G56, S57, I58, Y59, numbered according to the Kabat numbering system.
- the third polypeptide is (f1), (f2) or (f3).
- polypeptide (f1) An antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, or an antibody light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 47
- Polypeptide (f2) comprising antibody light chain variable domain having amino acid sequence of light chain CDR1, 2, 3 described in SEQ ID NO: 7, 8, 9 or antibody light chain variable having amino acid sequence described in SEQ ID NO: 47
- a polypeptide comprising a domain, numbered according to the Kabat numbering system, K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, A51, S52, R53, K54
- the fourth polypeptide is (g1), (g2) or (G3).
- G1 An antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 7, 8, 9 or an antibody light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 47
- a polypeptide comprising a domain, numbered according to the Kabat numbering system, K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, A51, S52, R53, K54
- a method for producing an Emicizumab variant comprising the following step (a): (A) replacing by one or more of the following (i) to (iii), wherein the numbering is according to the Kabat numbering system; (I) In the antibody light chain variable domain comprising the amino acid sequences of light chains CDR1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, K24, A25, S26, R27, N28, I29, E30, R31, Q32 Substitution in one or more amino acids selected from the group consisting of L33, A34, Q50, A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 (Ii) In the antibody heavy chain variable domain comprising the heavy chain CDR1, 2, 3 amino acid sequences set forth in SEQ ID NOs
- (I) In the antibody light chain variable domain comprising the amino acid sequences of light chains CDR1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, K24, A25, S26, R27, N28, I29, E30, R31, Q32 Substitution in one or more amino acids selected from the group consisting of L33, A34, Q50, A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 (Ii) In the antibody heavy chain variable domain comprising the heavy chain CDR1, 2, 3 amino acid sequences set forth in SEQ ID NOs: 1, 2, 3, Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a , S53, G54, Q55
- [14] to [26] are provided.
- [17] A nucleic acid encoding the antibody according to any one of [2], [3], [5], [7] to [9], [11] and [14] to [16]. [18] A vector into which the nucleic acid according to [17] has been inserted. [19] A cell comprising the nucleic acid according to [17] or the vector according to [17]. [20] [2], [3], [5], [7] to [9], [11] and [14] to [16], and a pharmaceutically acceptable carrier A pharmaceutical formulation comprising.
- the pharmaceutical preparation according to [20] which is a disease that develops and / or progresses due to a decrease or deficiency in the activity of a factor.
- the pharmaceutical preparation according to [21], wherein the disease that develops and / or progresses due to decreased or deficient activity of blood coagulation factor VIII and / or activated blood coagulation factor VIII is hemophilia A .
- a disease that develops and / or progresses due to a decrease or deficiency in the activity of blood coagulation factor VIII and / or activated blood coagulation factor VIII is caused by blood coagulation factor VIII and / or activated blood coagulation factor VIII
- the pharmaceutical agent according to [21], wherein the disease that develops and / or develops due to decreased or deficient activity of blood coagulation factor VIII and / or activated blood coagulation factor VIII is acquired hemophilia Formulation.
- antibody is used in the broadest sense and is not limited thereto as long as it exhibits a desired antigen-binding activity, but is not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (eg, Various antibody structures are included, including bispecific antibodies) and antibody fragments.
- Antibody fragment refers to a molecule other than the complete antibody, including a part of the complete antibody that binds to the antigen to which the complete antibody binds.
- Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab ′, Fab′-SH, F (ab ′) 2; diabody; linear antibody; single chain antibody molecule (eg, scFv And multispecific antibodies formed from antibody fragments.
- Class of an antibody refers to the type of constant domain or constant region provided in the heavy chain of the antibody.
- IgA immunoglobulin
- IgD immunoglobulin D
- IgE immunoglobulin D
- IgG immunoglobulin G
- IgM immunoglobulin M
- an “effective amount” of an agent refers to the amount at a required dose and over a required period of time that is effective to achieve a desired therapeutic or prophylactic result.
- Fc region is used herein to define the C-terminal region of an immunoglobulin heavy chain that includes at least a portion of the constant region.
- the term includes native sequence Fc regions and variant Fc regions.
- the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain.
- lysine (Lys447) or glycine-lysine (Gly446-Lys447) at the C-terminal of the Fc region may or may not be present.
- the numbering of amino acid residues in the Fc region or constant region is Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, follow the EU numbering system (also called EU index) described in MD 1991.
- “Framework” or “FR” refers to variable domain residues other than hypervariable region (HVR) residues.
- the FR of a variable domain usually consists of four FR domains: FR1, FR2, FR3, and FR4. Accordingly, HVR and FR sequences usually appear in VH (or VL) in the following order: FR1-H1 (L1) -FR2-H2 (L2) -FR3-H3 (L3) -FR4.
- full-length antibody “complete antibody”, and “total antibody” are used interchangeably herein and have a structure substantially similar to or defined herein.
- host cell refers to a cell (including the progeny of such a cell) into which a foreign nucleic acid has been introduced.
- Host cells include “transformants” and “transformed cells”, including the primary transformed cell and progeny derived from that cell regardless of the passage number.
- the progeny may not be completely identical in nucleic acid content with the parent cell, and may contain mutations. Also included herein are mutant progeny that have the same function or biological activity as was used when the original transformed cells were screened or selected.
- a “human antibody” is an antibody comprising an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell or an antibody derived from a non-human source using a human antibody repertoire or other human antibody coding sequence. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen binding residues.
- Humanized antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs.
- a humanized antibody comprises substantially all of at least one, typically two variable domains, in which all or substantially all HVRs (eg, CDRs) are non- It corresponds to that of a human antibody and all or substantially all FRs correspond to those of a human antibody.
- a humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody.
- a “humanized form” of an antibody (eg, a non-human antibody) refers to an antibody that has undergone humanization.
- hypervariable region or “HVR” as used herein is hypervariable in sequence (“complementarity determining region” or “CDR”) and / or structurally determined. Refers to each region of the variable domain of an antibody that forms a loop (“hypervariable loop”) and / or contains antigen contact residues (“antigen contact”). Usually, an antibody contains 6 HVRs: 3 in VH (H1, H2, H3) and 3 in VL (L1, L2, L3).
- Exemplary HVRs herein include the following: (a) at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) The resulting hypervariable loop (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)); (b) at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) The resulting CDR (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed.
- HVR residues and other residues in the variable domains are numbered herein according to Kabat et al., Supra.
- an “isolated” antibody is one that has been separated from its original environmental components.
- the antibody is, for example, by electrophoresis (eg, SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatograph (eg, ion exchange or reverse phase HPLC). Measured and purified to a purity greater than 95% or 99%.
- electrophoresis eg, SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis
- chromatograph eg, ion exchange or reverse phase HPLC
- isolated nucleic acid refers to a nucleic acid molecule that has been separated from its original environmental components.
- An isolated nucleic acid includes a nucleic acid molecule contained within a cell that normally contains the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or on a chromosome different from the original chromosomal location. Exists in position.
- the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies. That is, the individual antibodies that make up the population are mutated antibodies that can occur (eg, mutated antibodies that contain naturally occurring mutations, or mutated antibodies that occur during the production of monoclonal antibody preparations. Are present in the same amount and / or bind to the same epitope. In contrast to polyclonal antibody preparations that typically include different antibodies to different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is against a single determinant on the antigen.
- monoclonal indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
- monoclonal antibodies used in accordance with the present invention include, but are not limited to, hybridoma methods, recombinant DNA methods, phage display methods, transgenic animals containing all or part of a human immunoglobulin locus.
- Such methods and other exemplary methods for making monoclonal antibodies can be made by a variety of techniques, including methods utilizing the methods described herein.
- Natural antibodies refer to immunoglobulin molecules with various naturally occurring structures.
- a native IgG antibody is a heterotetrameric glycoprotein of approximately 150,000 daltons composed of two identical light chains and two identical heavy chains that are disulfide bonded.
- each heavy chain From the N-terminus to the C-terminus, each heavy chain has a variable region (VH) ⁇ ⁇ , also called variable heavy chain domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3).
- VH variable region
- VL variable region
- CL constant light chain
- the light chain of an antibody may be assigned to one of two types, called kappa ( ⁇ ) and lambda ( ⁇ ), based on the amino acid sequence of its constant domain.
- package insert is usually included in commercial packages of therapeutic products and includes information about indications, usage, dosages, administration methods, combination therapies, contraindications, and / or warnings regarding the use of such therapeutic products. Used to refer to instructions for use.
- pharmaceutical formulation refers to a preparation that is in a form such that the biological activity of the active ingredient contained therein can be effective and is unacceptable to the subject to which the formulation is administered. Refers to a preparation that does not contain additional toxic elements.
- “Pharmaceutically acceptable carrier” refers to an ingredient other than the active ingredient in a pharmaceutical preparation that is non-toxic to a subject.
- Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.
- FIX As used herein, the terms “FIX”, “FIXa” and “FX” are intended to include any mammal, including primates (eg, humans) and rodents (eg, mice and rats), unless otherwise indicated. Refers to any native FIX, FIXa and FX from any vertebrate source. The term encompasses FIX, FIXa, and FX that are not “full length” processed, as well as any form of FIX, FIXa, and FX that results from processing in a cell. The term also encompasses naturally occurring FIX, FIXa and FX variants, such as splice variants and allelic variants.
- treatment is clinical that is intended to alter the natural course of the individual being treated. Means intervention and can be carried out either for prevention or during the course of clinical pathology. Desirable effects of treatment include but are not limited to prevention of disease occurrence or recurrence, reduction of symptoms, attenuation of any direct or indirect pathological effects of the disease, prevention of metastasis, disease Includes reduced rate of progression, recovery or alleviation of disease state, and remission or improved prognosis.
- the antibodies of the invention are used to delay the onset of disease or slow the progression of disease.
- variable region refers to the heavy or light chain domain of an antibody involved in binding the antibody to an antigen.
- Natural antibody heavy and light chain variable domains (VH and VL, respectively) are typically similar, with each domain containing four conserved framework regions (FR) and three hypervariable regions (HVR). It has a structure. (See, for example, Kindt et al. Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)).
- One VH or VL domain will be sufficient to confer antigen binding specificity.
- antibodies that bind to a particular antigen may be isolated by screening complementary libraries of VL or VH domains, respectively, using VH or VL domains from antibodies that bind to the antigen. For example, see Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).
- vector refers to a nucleic acid molecule that can multiply another nucleic acid to which it has been linked.
- the term includes vectors as self-replicating nucleic acid structures and vectors that are integrated into the genome of the host cell into which it has been introduced. Certain vectors can provide for the expression of nucleic acids to which they are operably linked. Such vectors are also referred to herein as “expression vectors”.
- Emicizumab (ACE910), a bispecific antibody that binds to both FIX and / or FIXa and FX and replaces the function of FVIII, is described below.
- the bispecific antibody is a bispecific antibody in which a first polypeptide and a third polypeptide form a pair, and a second polypeptide and a fourth polypeptide form a pair.
- a heavy chain comprising the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 45
- the second polypeptide is an antibody heavy chain variable domain set forth in SEQ ID NO: 46.
- a bispecific antibody comprising a common L chain comprising the amino acid sequence of the antibody light chain variable domain set forth in SEQ ID NO: 47 as an H chain containing the amino acid sequence of the third polypeptide and the fourth polypeptide (Q499) -z121 / J327-z119 / L404-k).
- the bispecific antibody is a bispecific antibody in which a first polypeptide and a third polypeptide form a pair, and a second polypeptide and a fourth polypeptide form a pair.
- An H chain comprising the amino acid sequence set forth in SEQ ID NO: 10, a second polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 11, and a third polypeptide.
- a bispecific antibody (Q499-z121 / J327-z119 / L404-k) comprising a common L chain, wherein the polypeptide and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO: 12.
- Such an antibody can be obtained, for example, according to the method described in WO2005 / 035756, WO2006 / 109592, WO2012 / 067176, and the like.
- Emicizumab and ACE910 are synonymous in this specification.
- the “Emicizumab variant” in the present invention means a polypeptide or antibody in which at least one of the amino acid sequences of the heavy chain variable region or the light chain variable region of Emizicumab differs from the amino acid sequence of Emizicumab. All modifications are included as long as the object of the invention can be achieved. As one embodiment, even if all of the variable regions of some heavy chains or light chains of the variants are different from the variable regions of Emicizumab, they can be included in the variants of the present invention.
- Emicizumab variants are the following polypeptides and antibodies, but are not limited thereto.
- the present invention provides a polypeptide comprising an antibody light chain variable domain, wherein the antibody light chain variable domain has the amino acid sequence of light chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9.
- an antibody light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 47, wherein K24, A25, S26, R27, N28, I29, E30, R31, Q32, numbered according to the Kabat numbering system,
- One or more amino acids selected from the group consisting of L33, A34, Q50, A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 are cysteine Provided are polypeptides that are substituted with any amino acid other than.
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the amino acid sequences of the light chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 7, 8, and 9 are used.
- an antibody in which one or more amino acids are replaced with any amino acid other than cysteine.
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide and the third polypeptide form a pair, and the second polypeptide And the fourth polypeptide form a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 45, and the second polypeptide is the antibody heavy chain set forth in SEQ ID NO: 46
- the amino acid sequence of the variable domain, and the polypeptide of any one of the third polypeptide and the fourth polypeptide comprises the amino acid sequence of the antibody light chain variable domain set forth in SEQ ID NO: 47, and the other polypeptide Is an antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 7, 8, 9 or an antibody light chain having the amino acid sequence set forth in SEQ ID NO: 47
- the other polypeptide is numbered according to the Kabat numbering system, K24, A25,
- One or more amino acids selected from the group consisting of R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 are substituted with any amino acid other than cysteine.
- An antibody is provided.
- the present invention provides a polypeptide comprising an antibody heavy chain variable domain, wherein the antibody heavy chain variable domain has the amino acid sequence of heavy chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 1, 2, and 3.
- the antibody heavy chain variable domain has the amino acid sequence set forth in SEQ ID NO: 45, wherein Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a are numbered according to the Kabat numbering system.
- the present invention relates to a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the amino acid sequences of heavy chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 1, 2, and 3 are used.
- the present invention provides a polypeptide comprising an antibody heavy chain variable domain, wherein the antibody heavy chain variable domain has the amino acid sequence of heavy chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 4, 5, and 6.
- the present invention is a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the amino acid sequences of heavy chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 4, 5, and 6 are used.
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide and the third polypeptide form a pair, and the second polypeptide And the fourth polypeptide form a pair, the second polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 46, and the third polypeptide is the antibody light chain set forth in SEQ ID NO: 42
- the fourth polypeptide includes the amino acid sequence of the antibody light chain set forth in SEQ ID NO: 44, and the first polypeptide includes the heavy chain CDRs 1 and 2 set forth in SEQ ID NOs: 1, 2, and 3.
- One or more amino acids selected from the group consisting of T96, G97, R98, E99, Y100, G100a, G100b, G100c, W100d, Y100e, F100f, D101 and Y102 are substituted with any amino acid other than cysteine An antibody is provided.
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide and the third polypeptide form a pair, and the second polypeptide And the fourth polypeptide form a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain set forth in SEQ ID NO: 45, and the third polypeptide is the amino acid of the antibody light chain set forth in SEQ ID NO: 43
- the fourth polypeptide includes the amino acid sequence of the antibody light chain set forth in SEQ ID NO: 44, and the second polypeptide includes the heavy chain CDRs 1, 2, 3, set forth in SEQ ID NOs: 4, 5, and 6.
- D31, N32, N33, M34, D35, D50, I51, T52a, N52, R53, S54, G55, G56, S57, I58, Y59, N60, E61, E62, F63, Q64, D65, R95, K96 Provided is an antibody in which one or more amino acids selected from the group consisting of S97, Y98, G99, Y100, Y100a, L100b, D101 and E102 are substituted with any amino acid other than cysteine.
- the present invention provides a novel antibody light chain comprising any one of the following amino acid sequences selected from (a1) to (a6) and (b1) to (b23): .
- the light chain can be used as a light chain that replaces the common light chain contained in Emicizumab.
- the light chains (a1) to (a6) below are light chains that bind to FIX and / or FIXa, and the L chains (b1) to (b23) below are examples of light chains that bind to FX.
- the present invention provides the following antibody light chain variants.
- (c1) Antibody light chain having the amino acid sequence of SEQ ID NO: 42, which is a variant of the antibody light chain of SEQ ID NO: 13 (QAL187)
- (c2) Antibody L chain (QAL201) which is a variant of the antibody light chain described in SEQ ID NO: 13 and has the amino acid sequence described in SEQ ID NO: 43
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide (d) and the third polypeptide (f) form a pair.
- a second polypeptide (e) and a fourth polypeptide (g) form a pair, and each polypeptide provides an antibody as described below.
- the first polypeptide is (d1) or (d2).
- polypeptide (D1) An antibody heavy chain variable domain having the amino acid sequence of heavy chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 1, 2, 3 or an antibody heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 45
- Polypeptide (d2) containing antibody heavy chain variable domain having the amino acid sequence of heavy chain CDR1, 2, 3 as set forth in SEQ ID NO: 1, 2, 3 or antibody heavy chain variable having the amino acid sequence as set forth in SEQ ID NO: 45 A polypeptide comprising a domain, numbered according to the Kabat numbering system, Y31, Y32, D33, I34, Q35, S50, I51, S52, P52a, S53, G54, Q55, S56, T57, Y58, Y59 , R60, R61, E62, V63, K64, G65, R95, T96, G97, R98, E99, Y100, G100a, G100b, G100c, W100d, Y100e, F100
- the second polypeptide is (e1) or (e2).
- Polypeptide containing (e2) antibody heavy chain variable domain having amino acid sequence of heavy chain CDR1, 2, 3 described in SEQ ID NO: 4, 5, 6 or antibody heavy chain variable having amino acid sequence described in SEQ ID NO: 46 A polypeptide comprising a domain, numbered according to the Kabat numbering system, D31, N32, N33, M34, D35, D50, I51, T52a, N52, R53, S54, G55, G56, S57, I58, Y59 N60, E61, E62, F63, Q64, D65, R95, K96, S97, Y98, G99, Y100, Y100a, L100b, D101 and A polypeptide, wherein one or more amino acids selected from
- the third polypeptide is (f1), (f2) or (f3).
- (F1) An antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, or an antibody light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 47
- Polypeptide (f2) comprising antibody light chain variable domain having amino acid sequence of light chain CDR1, 2, 3 described in SEQ ID NO: 7, 8, 9 or antibody light chain variable having amino acid sequence described in SEQ ID NO: 47
- a polypeptide comprising a domain, numbered according to the Kabat numbering system, K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, A51, S52, R53, K54
- the fourth polypeptide is (g1), (g2) or (G3).
- G1 An antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, 3 set forth in SEQ ID NOs: 7, 8, 9 or an antibody light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 47
- a polypeptide comprising a domain, numbered according to the Kabat numbering system, K24, A25, S26, R27, N28, I29, E30, R31, Q32, L33, A34, Q50, A51, S52, R53, K54
- the present invention provides an amino acid residue at the following position in an antibody light chain variable domain having the amino acid sequence of light chain CDRs 1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, or SEQ ID NO: 47.
- a polypeptide or antibody in which at least one of the amino acid residues at the following positions in the antibody light chain variable domain described is any of the amino acid residues described below.
- an amino acid residue different from the antibody light chain variable domain having the amino acid sequence of light chain CDR1, 2, 3 described in SEQ ID NOs: 7, 8, 9, or the antibody described in SEQ ID NO: 47 The number of amino acid residues different from the light chain variable domain may be one or more, preferably 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and more preferably 21 or more.
- the replacement position indicates a position numbered according to the Kabat numbering system.
- K24 A, I, L, M, P, V, G, N, Q, S, T, D, E, H, R, F, W, Y A25: I, L, M, P, V, G, N, Q, S, T, D, E, H, K, R, F, W, Y S26: A, I, L, M, P, V, G, N, Q, T, D, E, H, K, R, F, W, Y R27: A, I, L, M, P, V, G, N, Q, S, T, D, E, H, K, F, W, Y N28: A, I, L, M, P, V, G, Q, S, T, D, E, H, K, R, F, W, Y I29: A, L, M, P, V, G, N, Q, S, T, D, E, H, K, R, F, W, Y I29: A, L, M, P, V, G, N, Q, S,
- the present invention relates to an amino acid residue at the following position in an antibody heavy chain variable domain having the amino acid sequence of heavy chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 1, 2, and 3, or SEQ ID NO: 45
- a polypeptide or antibody in which at least one of the amino acid residues at the following positions in the antibody heavy chain variable domain described above is any of the amino acid residues described below.
- an amino acid residue different from the antibody heavy chain variable domain having the amino acid sequence of the heavy chain CDRs 1, 2, and 3 shown in SEQ ID NOs: 1, 2, and 3, or the anti-antibody shown in SEQ ID NO: 45
- the number of amino acid residues different from the weight chain variable domain may be one or more, preferably 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and more preferably 21 or more.
- the replacement position indicates a position numbered according to the Kabat numbering system.
- Y31 A, I, L, M, P, V, G, N, Q, S, T, D, E, H, K, R, F, W Y32: A, I, L, M, P, V, G, N, Q, S, T, D, E, H, K, R, F, W D33: A, I, L, M, P, V, G, N, Q, S, T, E, H, K, R, F, W, Y I34: A, L, M, P, V, G, N, Q, S, T, D, E, H, K, R, F, W, Y Q35: A, I, L, M, P, V, G, N, S, T, D, E, H, K, R, F, W, Y S50: A, I, L, M, P, V, G, N, Q, T, D, E, H, K, R, F, W, Y I51: A, L, M, P, V, G, N, Q, T,
- the present invention relates to an amino acid residue at the following position in an antibody heavy chain variable domain having the amino acid sequence of heavy chain CDR1, 2, 3 described in SEQ ID NOs: 4, 5, 6, or SEQ ID NO: 46.
- a polypeptide or antibody in which at least one of the amino acid residues at the following positions in the antibody heavy chain variable domain described above is any of the amino acid residues described below.
- the number of amino acid residues different from the weight chain variable domain may be one or more, preferably 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and more preferably 21 or more.
- the replacement position indicates a position numbered according to the Kabat numbering system.
- D31 A, I, L, M, P, V, G, N, Q, S, T, E, H, K, R, F, W, Y N32: A, I, L, M, P, V, G, Q, S, T, D, E, H, K, R, F, W, Y N33: A, I, L, M, P, V, G, Q, S, T, D, E, H, K, R, F, W, Y M34: A, I, L, P, V, G, N, Q, S, T, D, E, H, K, R, F, W, Y D35: A, I, L, M, P, V, G, N, Q, S, T, E, H, K, R, F, W, Y D50: A, I, L, M, P, V, G, N, Q, S, T, E, H, K, R, F, W, Y D50: A, I, L, M, P, V, G, N, Q,
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide and the third polypeptide are paired, and the second polypeptide
- the fourth polypeptide forms a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 45, and the second polypeptide is the antibody heavy chain variable set forth in SEQ ID NO: 46.
- the amino acid sequence of the domain, the fourth polypeptide includes the amino acid sequence of the antibody light chain variable domain set forth in SEQ ID NO: 47, and the third polypeptide is any one selected from (a1) to (a6) above An antibody comprising an antibody light chain comprising an amino acid sequence is provided.
- the present invention provides a bispecific antibody that recognizes FIX and / or FIXa and FX, wherein the first polypeptide and the third polypeptide are paired, and the second polypeptide And the fourth polypeptide form a pair, the first polypeptide is the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 45, and the third polypeptide is the antibody light chain set forth in SEQ ID NO: 47
- the amino acid sequence of the variable domain the second polypeptide includes the amino acid sequence of the antibody heavy chain variable domain set forth in SEQ ID NO: 46, respectively, and the fourth polypeptide is any one selected from (b1) to (b23) above
- An antibody comprising an antibody light chain comprising the amino acid sequence of:
- the invention provides variants of the polypeptides and antibodies provided herein.
- the variants provided herein include deletion of amino acid residues in the amino acid sequence and / or insertion of amino acid residues in the amino acid sequence, and / or amino acids in the amino acid sequence.
- Variants in which residue substitutions have been made are included. Given that the final construct has improved FVIII cofactor function replacement activity, any combination of deletions, insertions, and substitutions can be made to arrive at the final construct.
- FIX (a) means FIX and / or FIXa.
- the antibodies provided herein are antibody fragments.
- Antibody fragments include, but are not limited to, Fab, Fab ′, Fab′-SH, F (ab ′) 2 , Fv, and scFv fragments, as well as other fragments described below.
- Fab fragment antigen
- Fab′ fragment antigen binding domain
- a diabody is an antibody fragment with two antigen binding sites, which may be bivalent or bispecific.
- a single domain antibody is an antibody fragment comprising all or part of an antibody heavy chain variable domain or all or part of a light chain variable domain.
- the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, eg, US Pat. No. 6,248,516 B1).
- Antibody fragments include, but are not limited to, various forms of proteolytic digestion of complete antibodies, as described herein, including production by recombinant host cells (eg, E. coli or phage). It can be made by the method of.
- the antibodies provided herein are human antibodies.
- Human antibodies can be produced by various techniques known in the art. Human antibodies are reviewed in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).
- Human antibodies may be prepared by administering an immunogen to a transgenic animal that has been modified to produce fully human antibodies or fully antibodies with human variable regions in response to an antigen challenge (load).
- Such animals typically include all or part of a human immunoglobulin locus, which all or part of the human immunoglobulin locus replaces the endogenous immunoglobulin locus or is extrachromosomal or It exists in a state of being randomly incorporated in the chromosome of the animal. In such transgenic mice, the endogenous immunoglobulin locus is usually inactivated.
- Human antibodies can also be made by methods based on hybridomas. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have already been described. (Eg, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp.51-63 (Marcel Dekker, Inc., New York, 1987); , J. Immunol., 147: 86 (1991).) Human antibodies generated via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557- It is stated in 3562 (2006). Additional methods include, for example, US Pat.
- Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences can then be combined with the desired human constant domain. A technique for selecting a human antibody from an antibody library is described below.
- Antibodies of the present invention may be isolated by screening combinatorial libraries for antibodies with one or more desired activities. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with the desired binding properties. Such methods are reviewed in Hoogenboom et al. In Methods in Molecular Biology 178: 1-37 (O'Brien et al., Ed., Human Press, Totowa, NJ, 2001), and further e.g. McCafferty et al., Nature 348: 552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol.
- phage display methods repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR) and randomly recombined in a phage library, which is linked to a Winter library. ., Ann. Rev. Immunol., 12: 433-455 (1994) ⁇ ⁇ can be screened for antigen-binding phages. Phages typically display antibody fragments, either as single chain Fv (scFv) fragments or as Fab fragments. Libraries from immunized sources provide high affinity antibodies to the immunogen without the need to construct hybridomas.
- scFv single chain Fv
- the na ⁇ ve repertoire can be cloned (eg, from human) without extensive immunization and without immunization. It is also possible to provide a single source of antibodies to self antigens.
- the na ⁇ ve library was used to clone the pre-rearranged V-gene segment from stem cells as described in Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992) It can also be made synthetically by using PCR primers that encode the region and contain random sequences to achieve reconstitution in vitro.
- Patent documents describing human antibody phage libraries include, for example: US Pat. No.
- an antibody or antibody fragment isolated from a human antibody library is regarded as a human antibody or a human antibody fragment.
- Multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (Milstein and Cuello, Nature 305: 537 (1983), WO 93/08829, and Traunecker et al., EMBO J. 10: 3655 (1991)), and the knob-in-hole technology (see, eg, US Pat. No. 5,731,168).
- Multispecific antibodies can manipulate electrostatic steering effects to create Fc heterodimeric molecules (WO2009 / 089004A1); bridge two or more antibodies or fragments (US patents) No.
- antibody variants having one or more amino acid substitutions are provided.
- Target sites for substitutional mutagenesis include HVR and FR.
- Conservative substitutions are shown under the heading “Preferred substitutions” in Table 1. More substantial changes are provided under the heading “Exemplary substitutions” in Table 1 and are detailed below with reference to the class of amino acid side chains.
- Amino acid substitutions may be introduced into the antibody of interest, and the product is desired, eg, retained / improved antigen binding, decreased immunogenicity, or improved FVIII cofactor function surrogate activity, ADCC or CDC. May be screened for activity.
- Amino acids can be grouped according to common side chain properties: (1) Hydrophobicity: norleucine, methionine (Met), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile); (2) Neutral hydrophilicity: cysteine (Cys), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln); (3) Acidity: Aspartic acid (Asp), glutamic acid (Glu); (4) Basicity: histidine (His), lysine (Lys), arginine (Arg); (5) Residues that affect chain orientation: Glycine (Gly), Proline (Pro); (6) Aromaticity: Tryptophan (Trp), Tyrosine (Tyr), Phenylalanine (Phe). Non-conservative substitutions refer to exchanging one member of these classes for another class.
- substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (eg, a humanized or human antibody).
- a parent antibody eg, a humanized or human antibody.
- the resulting mutants selected for further study are modified (eg, improved) in certain biological properties compared to the parent antibody (eg, increased FVIII cofactor function replacement activity or Affinity, reduced immunogenicity) and / or substantially retain certain biological properties of the parent antibody.
- substitutions, insertions, or deletions can be made in one or more HVRs as long as such modification does not substantially reduce the ability of the antibody to bind to the antigen.
- conservative modifications eg, conservative substitutions as provided herein
- FVIII cofactor function surrogate activity can be made.
- a useful method for identifying antibody residues or regions that can be targeted for mutagenesis is described by Cunningham and Wells (1989) Science, 244: 1081-1085, ⁇ Alanine scanning mutagenesis ''. It is what is called.
- a residue or group of target residues eg, charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid
- neutral or negatively charged amino acids eg, alanine or Polyalanine
- the crystal structure of the antigen-antibody complex may be analyzed to identify contact points between the antibody and the antigen. Such contact residues and neighboring residues may be targeted as substitution candidates or excluded from substitution candidates. Variants can be screened to determine if they contain the desired property.
- Amino acid sequence insertions are within the length of a polypeptide comprising from 1 to 100 residues or more at the amino and / or carboxyl terminus. Includes fusion.
- Examples of terminal insertions include antibodies with a methionyl residue at the N-terminus.
- Other insertional variants of the antibody molecule include those in which the N- or C-terminus of the antibody is fused to an enzyme (eg, for ADEPT) or a polypeptide that increases the plasma half-life of the antibody.
- the antibodies provided herein have been modified to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to the antibody can be conveniently accomplished by altering the amino acid sequence to create or remove one or more glycosylation sites.
- the carbohydrate added thereto may be modified.
- Natural-type antibodies produced by mammalian cells typically contain a branched biantennary oligosaccharide, which is usually added by N-linkage to Asn297 in the CH2 domain of the Fc region.
- Oligosaccharides include, for example, various carbohydrates such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose added to GlcNAc in the “stem” of the biantennary oligosaccharide structure.
- modification of oligosaccharides in the antibodies of the invention may be made to create antibody variants with certain improved properties.
- antibody variants having carbohydrate structures that lack fucose added (directly or indirectly) to the Fc region are provided.
- the amount of fucose in such antibodies can be 1% -80%, 1% -65%, 5% -65% or 20% -40%.
- the amount of fucose is the sum of all sugar structures (eg, complex, hybrid, and high mannose structures) added to Asn297 as measured by MALDI-TOF mass spectrometry as described, for example, in WO2008 / 077546 In contrast, it is determined by calculating the average amount of fucose in the sugar chain in Asn297.
- Asn297 represents an asparagine residue located around position 297 in the Fc region (EU numbering of Fc region residues). However, due to the slight sequence diversity between the antibodies, Asn297 may be located ⁇ 3 amino acids upstream or downstream of position 297, ie between positions 294-300. Such fucosylated mutants may have improved ADCC function. See, for example, US Patent Application Publication No. 2003/0157108 (Presta, L.); 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd).
- an antibody variant having a bisected oligosaccharide for example, a biantennary oligosaccharide added to the Fc region of an antibody is bisected by GlcNAc.
- Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO2003 / 011878 (Jean-Mairet et al.); US Pat. No. 6,602,684 (Umana et al); and US2005 / 0123546 (Umana et al). Yes.
- antibody variants having at least one galactose residue in the oligosaccharide added to the Fc region Such antibody variants may have improved CDC function.
- Such antibody variants are described, for example, in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S); and WO 1999/22764 (Raju, S).
- Fc region variants In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of the antibodies provided herein, thereby generating Fc region variants.
- An Fc region variant may comprise a human Fc region sequence (eg, a human IgG1, IgG2, IgG3, or IgG4 Fc region) comprising an amino acid modification (eg, substitution) at one or more amino acid positions.
- antibody variants with some, but not all, effector functions are also within the scope of the present invention, which effector functions with respect to its in vivo half-life, Certain effector functions (such as complement and ADCC) are good candidates for application when they are unnecessary or harmful.
- In vitro sputum and / or in vivo sputum cytotoxicity measurements can be performed to confirm a decrease / deficiency of CDC and / or ADCC activity.
- Fc receptor (FcR) binding measurements can be performed to ensure that antibodies lack Fc ⁇ R binding (and thus are more likely to lack ADCC activity) while maintaining FcRn binding ability.
- NK cells which are primary cells that mediate ADCC, express only Fc ⁇ RIII, whereas monocytes express Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII.
- the expression of FcR on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9: 457-492 (1991).
- in vitro assays for assessing ADCC activity of a molecule of interest include US Pat. No. 5,500,362 (eg, Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA). 83: 7059-7063 (1986)) and Hellstrom, I et al., Proc.
- non-radioactive assays may be used (eg, ACT1 TM non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, CA); and CytoTox 96® non-radioactive cytotoxicity assays (see Promega, Madison, WI). Effector cells useful for such measurement methods include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells.
- PBMC peripheral blood mononuclear cells
- NK natural killer
- the ADCC activity of the molecule of interest is assessed in vivo in an animal model as described, for example, in Clynes et al. Proc. Nat'l Acad. Sci. USA 95: 652-656 (1998). May be.
- C1q binding measurements may be performed to confirm that the antibody cannot bind to C1q and thus lacks CDC activity. See, for example, the C1q and C3c binding ELISA in WO2006 / 029879 and WO2005 / 100402.
- CDC measurements may also be performed to assess complement activation (eg, Gazzano-Santoro et al., J. Immunol.
- Antibodies with reduced effector function include those with one or more substitutions of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (US Pat. No. 6,737,056).
- Fc variants include amino acid positions 265, 269, 270, 297, and 327, including so-called “DANA” Fc variants (US Pat. No. 7,332,581) with substitution of residues 265 and 297 to alanine.
- DANA DANA
- modified ie, increased
- CDC complement dependent cytotoxicity selection
- FcRn maternal IgGs to the fetus
- Kim et al. J. Antibodies with increased binding to Immunol. 24: 249 (1994)
- FcRn maternal IgGs to the fetus
- These antibodies comprise an Fc region with one or more substitutions therein that increase the binding of the Fc region to FcRn.
- Such Fc variants include Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382 , 413, 424, 428, 434 or 436 with substitutions at one or more (eg, substitution of Fc region residue 434 (US Pat. No. 7,371,826)).
- Fc region mutants in which binding to rheumatoid factor is suppressed can be used for the antibody of the present invention, and such mutants are described in WO2013 / 046704, for example, Q438R / S440E, Q438R / S440D, Mention may be made of mutants having a combination of mutations indicated by Q438K / S440E or Q438K / S440D in both H chains.
- Fc region mutants having enhanced binding to FcRn and suppressed binding to rheumatoid factor can be used for the antibody of the present invention. More specifically, Fc region mutants having the combination of mutations shown in any one of the following a) to f) in both H chains can be mentioned.
- N434A / Y436T / Q438R / S440E b) N434A / Y436V / Q438R / S440E c) M428L / N434A / Y436T / Q438R / S440E d) M428L / N434A / Y436V / Q438R / S440E e) M428L / N434A / Q438R / S440E f) N434A / Q438R / S440E
- Fc region variants see Duncan & Winter, Nature 322: 738-40 (1988); US Pat. No. 5,648,260; US Pat. No. 5,624,821; and WO94 / 29351.
- the antibodies provided herein may be further modified to include additional non-protein moieties known in the art and readily available. Suitable moieties for antibody derivatization include, but are not limited to, water soluble polymers.
- Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly 1,3 Dioxolane, poly1,3,6, trioxane, ethylene / maleic anhydride copolymer, polyamino acid (either homopolymer or random copolymer), and dextran or poly (n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymer, Polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (eg, glycerol), polyvinyl alcohol, and mixtures thereof.
- PEG polyethylene glycol
- ethylene glycol / propylene glycol copolymers carboxymethyl cellulose
- dextran polyvinyl alcohol
- polyvinyl pyrrolidone poly
- Polyethylene glycol propionaldehyde may be advantageous in manufacturing due to its water stability.
- the polymer may have any molecular weight and may or may not be branched.
- the number of polymers added to the antibody can vary, and if more than one polymer is added, they can be the same molecule or different molecules. In general, the number and / or type of polymers used for derivatization is not limited to these, but is specific to the antibody's particular characteristics or function to be improved, under conditions where the antibody derivative is under defined conditions. It can be determined based on considerations such as whether or not to use in therapy.
- Antibodies can be produced using recombinant methods and configurations, for example, as described in US Pat. No. 4,816,567.
- an isolated nucleic acid encoding an anti-FIX (a) / FX bispecific antibody described herein is provided.
- Such a nucleic acid may encode an amino acid sequence comprising an antibody VL and / or an amino acid sequence comprising a VH (eg, an antibody light and / or heavy chain).
- one or more vectors eg, expression vectors
- host cells comprising such nucleic acids are provided.
- the host cell comprises (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising the antibody VL and an amino acid sequence comprising the antibody VH, or (2) an amino acid comprising the antibody VL.
- a first vector comprising a nucleic acid encoding sequence and a second vector comprising a nucleic acid encoding an amino acid sequence comprising the antibody VH are included (eg, transformed).
- the host cell is eukaryotic (eg, Chinese hamster ovary (CHO) cells) or lymphoid cells (eg, Y0, NS0, Sp2 / 0 cells)).
- culturing host cells comprising a nucleic acid encoding the antibody as described above under conditions suitable for expression of the anti-FIX (a) / FX bispecific antibody, and optionally, the antibody
- a method is provided for making an anti-FIX (a) / FX bispecific antibody comprising recovering from a host cell (or host cell culture medium).
- nucleic acids encoding the antibodies are isolated for further cloning and / or expression in host cells. In one or more vectors. Such nucleic acids will be readily isolated and sequenced using conventional procedures (eg, oligonucleotide probes that can specifically bind to the genes encoding the antibody heavy and light chains). By using).
- Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells as described herein.
- antibodies may be produced in bacteria, particularly where glycosylation and Fc effector function are not required. See, eg, US Pat. Nos. 5,648,237, 5,789,199, and 5,840,523 for expression of antibody fragments and polypeptides in bacteria. (In addition, see also Charlton, Methods Molecular Biology, Vol.BK248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp.245-254, which describes the expression of antibody fragments in E. coli. ) After expression, the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.
- true species such as filamentous fungi or yeast, including fungal and yeast strains in which the glycosylation pathway is “humanized”, resulting in the production of antibodies with partial or complete human glycosylation patterns.
- Nuclear microorganisms are suitable cloning or expression hosts for antibody-encoding vectors. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004) and Li et al., Nat. Biotech. 24: 210-215 (2006).
- invertebrate and vertebrate organisms are also suitable host cells for the expression of glycosylated antibodies.
- invertebrate cells include plant and insect cells.
- a number of baculovirus strains have been identified that are used for mating with insect cells, particularly for transformation of Spodoptera frugiperda cells.
- Plant cell cultures can also be used as hosts. See, for example, US Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (described PLANTIBODIES TM technology for producing antibodies in transgenic plants).
- Vertebrate cells can also be used as hosts.
- mammalian cell lines adapted to grow in suspension will be useful.
- Other examples of useful mammalian host cell lines include monkey kidney CV1 strain (COS-7) transformed with SV40; human embryonic kidney strain (Graham et al., J. Gen Virol. 36:59 (1977 293 or 293 cells as described in); pup hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells as described in Mather, Biol. Reprod.
- monkey kidney cell (CV1 ); African green monkey kidney cell line (VERO-76); Human cervical cancer cell line (HELA); Canine kidney cell line (MDCK); Buffalo rat liver cell line (BRL 3A); Human lung cell line (W138); Hep G2); mouse breast cancer (MMT 060562); TRI cells (for example, described in Mather et al., Annals NY Acad. Sci. 383: 44-68 (1982)); MRC5 cells; and FS4 cells.
- Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci.
- Y0 Includes myeloma cell lines such as NS0 and Sp2 / 0.
- myeloma cell lines such as NS0 and Sp2 / 0.
- Measurement method The anti-FIX (a) / FX bispecific antibodies provided herein are identified, screened, or have physical / chemical properties and / or by various assays known in the art. Or it may be revealed about biological activity.
- the antibodies of the present invention are tested for their antigen binding activity by known methods such as ELISA, Western blot, and the like.
- a assay for identifying that of an anti-FIX (a) / FX bispecific antibody having biological activity is provided.
- Biological activity may include, for example, FVIII cofactor function surrogate activity.
- antibodies having such biological activity in vivo and / or in vitro are provided.
- the antibodies of the invention are tested for such biological activity.
- FVIII cofactor function substitution activity In the present invention, “FVIII cofactor function substitution activity”, “FVIII substitution activity” and “activity substitution for FVIII function” are used synonymously to recognize FIX and / or FIXa and FX and promote the activation of FX. It means activity that promotes FXa production.
- FVIII cofactor function alternative activity in the present invention means, for example, an activity calculated from an FX activation reaction test using FIXa using a colorimetric method and a thrombin generation test using hemophilia A plasma. . More specifically, it can be confirmed by evaluating the antibody of the present invention with a measurement system comprising, for example, FIXa, FX, synthetic substrate S-2222 (FXa synthetic substrate), and phospholipid. This measurement system correlates with the severity and clinical symptoms of hemophilia A cases (Rosen S, Andersson M, Blomba ⁇ ck M et al. Clinical applications of a chromogenic substrate method for determination of FVIII activity. Thromb Haemost 1985; 54: 811-23).
- Anti-FIX (a) / FX bispecific antibody pharmaceutical formulations described herein can be prepared by administering an antibody having a desired purity to one or more optional pharmaceutically acceptable carriers ( Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) and is prepared in the form of a lyophilized formulation or an aqueous solution.
- Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations employed, including but not limited to the following: phosphate, citric acid Buffers such as acid salts and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, Or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); small molecule (less than about 10 residues) polypeptide; serum albumin, gelatin, or Proteins such as immunoglobulins; polyvinyl Hydrophilic polymers such as loridone; amino acids such as glycine, glutamine, asparagine, histidine, argin
- Exemplary pharmaceutically acceptable carriers herein further include a soluble neutral active hyaluronidase glycoprotein (sHASEGP) (eg, rHuPH20 (HYLENEX®, Baxter International, Inc.) PH-20 hyaluronidase glycoprotein) and other interstitial drug dispersants.
- sHASEGP soluble neutral active hyaluronidase glycoprotein
- rHuPH20 HYLENEX®, Baxter International, Inc.
- PH-20 hyaluronidase glycoprotein interstitial drug dispersants.
- sHASEGPs and their methods of use including rHuPH20
- sHASEGP is combined with one or more additional glycosaminoglycanases such as chondroitinase.
- the active ingredients are incorporated into microcapsules prepared by, for example, droplet formation (coacervation) techniques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatin microcapsules and poly (methyl methacrylate) microcapsules, respectively).
- it may be incorporated into colloidal drug delivery systems (eg, liposomes, albumin spherules, microemulsions, nanoparticles, and nanocapsules) or macroemulsions.
- colloidal drug delivery systems eg, liposomes, albumin spherules, microemulsions, nanoparticles, and nanocapsules
- Such a technique is disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).
- sustained release preparations may be prepared.
- suitable examples of sustained release formulations include a semi-permeable matrix of solid hydrophobic polymer containing antibodies, which matrix is in the form of shaped articles such as films or microcapsules.
- the preparations used for in vivo administration are usually sterile. Aseptic conditions are easily achieved, for example, by filtration through sterile filtration membranes.
- anti-FIX (a) / FX bispecific antibodies may be used in therapeutic methods.
- anti-FIX (a) / FX bispecific antibodies are provided for use as pharmaceuticals.
- anti-FIX (a) / FX bispecific antibodies are provided for use in the treatment of bleeding, bleeding-related diseases, or diseases resulting from bleeding.
- anti-FIX (a) / FX bispecific antibodies are provided for use in therapeutic methods.
- the present invention provides a method of treating an individual having bleeding, a disease involving bleeding, or a disease caused by bleeding, wherein the individual is treated with anti-FIX (a) / FX bispecific antibody.
- An anti-FIX (a) / FX bispecific antibody is provided for use in a method comprising administering an amount.
- the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent (eg, as described below).
- the present invention provides anti-FIX (a) / FX bispecific antibodies for use in surrogate FVIII function.
- the present invention is a method of substituting FVIII function in an individual, wherein an effective amount of an anti-FIX (a) / FX bispecific antibody is administered to the individual in order to replace FVIII function.
- An anti-FIX (a) / FX bispecific antibody is provided for use in a method comprising the steps of:
- An “individual” according to any of the above embodiments is preferably a human.
- the antibody of the present invention is an agent effective against a disease caused by a decrease in the activity (function) of the cofactor.
- diseases include bleeding, bleeding-related diseases, and diseases caused by bleeding, and preferably the onset is caused by a decrease or deficiency in the activity of FVIII and / or activated blood coagulation factor VIII (FVIIIa). And / or developing disease.
- diseases include hemophilia A, diseases in which inhibitors for FVIII / FVIIIa have appeared, acquired hemophilia, von Willebrand disease and the like, but are not particularly limited to these diseases.
- the present invention provides the use of an anti-FIX (a) / FX bispecific antibody in the manufacture or preparation of a medicament.
- the medicament is for the treatment of bleeding, bleeding-related diseases, or diseases resulting from bleeding.
- the medicament is a method of treating bleeding, a disease associated with bleeding, or a disease caused by bleeding, wherein an effective amount of the drug is administered to an individual having bleeding, a disease involving bleeding, or a disease caused by bleeding.
- the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent (eg, as described below).
- the medicament is for replacement of FVIII function.
- the medicament, a method for replacing the FVIII functions in an individual is for use in a method comprising administering an effective amount of a medicament to the individual in order to substitute the FVIII function.
- An “individual” according to any of the above aspects may be a human.
- the present invention provides a method for treating bleeding, diseases associated with bleeding, or diseases resulting from bleeding.
- the method comprises administering an effective amount of an anti-FIX (a) / FX bispecific antibody to an individual having such bleeding, a disease associated with bleeding, or a disease resulting from bleeding.
- the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent (as described below).
- An “individual” according to any of the above aspects may be a human.
- the present invention substitutes for FVIII function in an individual Providing a method for In one embodiment, the method comprises administering to the individual an effective amount of an anti-FIX (a) / FX bispecific antibody to replace FVIII function.
- an “individual” is a human.
- the present invention provides a pharmaceutical formulation comprising any of the anti-FIX (a) / FX bispecific antibodies provided herein (eg, any of the therapeutic methods described above).
- the pharmaceutical formulation comprises any of the anti-FIX (a) / FX bispecific antibodies provided herein and a pharmaceutically acceptable carrier.
- the pharmaceutical formulation comprises any of the anti-FIX (a) / FX bispecific antibodies provided herein and at least one additional therapeutic agent (eg, as described below). Including.
- the antibody of the present invention can be used in therapy either alone or in combination with other agents.
- an antibody of the invention may be co-administered with at least one additional therapeutic agent.
- the additional therapeutic agent is an FVIII formulation.
- Combination therapy as described above includes combination administration (two or more therapeutic agents included in the same or separate formulations) and individual administration, where administration of the antibody of the present invention is an additional treatment. Prior to, simultaneously with and / or subsequent to administration of the agent.
- the administration of the anti-FIX (a) / FX bispecific antibody and the administration of the additional therapeutic agent are within about one month, or within about 1, 2, or 3 weeks of each other, or about 1, 2 Done within 3, 4, 5, or 6 days.
- the antibodies (and any additional therapeutic agent) of the invention may be administered by any suitable means, including parenteral, pulmonary, and nasal administration, and intralesional administration if desired for local treatment.
- Parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
- Dosing can be by any suitable route, such as by injection, such as intravenous or subcutaneous injection, depending in part on whether administration is short or long term.
- Various dosing schedules are within the scope of this specification, including, but not limited to, single doses or repeated doses over various time points, bolus doses, and pulse infusions.
- the antibody of the present invention is formulated, dosed and administered in a manner consistent with good medical practice. Factors to be considered in this regard are the specific disorder being treated, the particular mammal being treated, the clinical symptoms of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration Schedule, and other factors known to healthcare professionals.
- the antibody need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question.
- the effective amount of such other agents depends on the amount of antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. These are usually at the same dose and route of administration as described herein, or at about 1 to 99% of the doses described herein, or any dose determined empirically / clinically appropriate. And in any route.
- an appropriate dose of the antibody of the present invention (when used alone or in combination with one or more other additional therapeutic agents) will determine the type of disease being treated, It will depend on the type, severity and course of the disease, whether the antibody is administered for prophylactic or therapeutic purposes, drug history, patient clinical history and response to antibodies, and the discretion of the attending physician .
- the antibody is suitably administered to the patient at one time or over a series of treatments.
- about 1 ⁇ g / kg to 15 mg / kg (eg, 0.1 mg / kg to 10 mg) antibody may be the first candidate dose for administration to a patient.
- One typical daily dose may vary from about 1 ⁇ g / kg to over 100 mg / kg, depending on the factors described above. For repeated administrations over several days or longer, depending on the situation, treatment is usually maintained until a desired suppression of disease symptoms occurs.
- One exemplary dose of antibody is in the range of about 0.05 mg / kg to about 10 mg / kg.
- one or more doses (or any combination thereof) of about 0.5 mg / kg, 2.0 mg / kg, 4.0 mg / kg, or 10 mg / kg may be administered to the patient.
- Such doses may be administered intermittently, such as every week or every 3 weeks (eg, so that the patient receives about 2 to about 20, or such as about 6 doses of antibody).
- One or more low doses may be administered after the high initial loading dose. The progress of this therapy is easily monitored by conventional techniques and measurements.
- products that include equipment useful for the treatment, prevention, and / or diagnosis of the disorders described above.
- the product includes a container and a label on the container or package insert attached to the container.
- Preferred containers include, for example, bottles, vials, syringes, IV solution bags and the like.
- the containers may be formed from various materials such as glass and plastic.
- the container may hold the composition alone, in combination with another composition that is effective for the treatment, prevention, and / or diagnosis of symptoms, and with a sterile access port.
- the container may be a solution bag or vial for intravenous administration having a stopper that can be pierced by a hypodermic needle).
- At least one active ingredient in the composition is an antibody of the invention.
- the label or package insert indicates that the composition is to be used to treat the selected condition.
- the product further comprises: (a) a first container with a composition comprising an antibody of the present invention contained therein; and (b) a second container, A second container may be included with the composition containing an additional cytotoxic agent or otherwise therapeutic agent contained therein.
- the product in this aspect of the invention may further include a package insert indicating that the composition can be used to treat a particular condition.
- the product further comprises a second (or pharmaceutically acceptable buffer, such as water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution.
- BWFI water for injection
- phosphate buffered saline such as phosphate buffered saline, Ringer's solution, and dextrose solution.
- a third) container may be included.
- Other buffers, diluents, filters, needles, and syringes may further include equipment desirable from other commercial
- the present invention is a method for producing an Emicizumab variant, which comprises the following step (a). (A) replacing by one or more of the following (i) to (iii), wherein the numbering is according to the Kabat numbering system; (I) In the antibody light chain variable domain comprising the amino acid sequences of light chains CDR1, 2, and 3 set forth in SEQ ID NOs: 7, 8, and 9, K24, A25, S26, R27, N28, I29, E30, R31, Q32 Substitution in one or more amino acids selected from the group consisting of L33, A34, Q50, A51, S52, R53, K54, E55, S56, Q89, Q90, Y91, S92, D93, P94, P95, L96 and T97 (Ii) In the antibody heavy chain variable domain comprising the heavy chain CDR1, 2, 3 amino acid sequences set forth in SEQ ID NOs: 1, 2, 3, Y31, Y32, D33, I34, Q35, S50,
- the number of substitution sites may be one or more, and a combination of (i) and (ii), a combination of (i) and (iii), a combination of (ii) and (iii), (i) and (ii) And a combination of (iii).
- the present invention is a method for isolating an Emicizumab variant, comprising the following steps (a) to (c).
- the number of substitution sites may be one or more, and a combination of (i) and (ii), a combination of (i) and (iii), a combination of (ii) and (iii), (i) and (ii) And a combination of (iii).
- ACE910 is a humanized IgG4 antibody consisting of anti-FIX (a) and anti-FX showing FVIII cofactor function replacement activity, and FIX (a) And FX consists of two types of heavy chains (Q499 and J327) and a common L chain (L404) (heavy chain SEQ ID NO: 45 and 46, light chain SEQ ID NO: 47).
- L404 common L chain
- the present inventors comprehensively introduce amino acid modifications to L404 by methods known to those skilled in the art, such as PCR for mutagenesis, and conduct large-scale screening for FVIII cofactor function substitution activity.
- the present inventors have found an amino acid substitution of the L chain that improves the FVIII cofactor function replacement activity of ACE910.
- the L chain on the anti-FIX (a) antibody side of ACE910 (heavy chain SEQ ID NO: 45, light chain SEQ ID NO: 47) is fixed to L404 (SEQ ID NO: 47), and the anti-FX antibody side of ACE910 (heavy chain SEQ ID NO: 47) : 46, light chain SEQ ID NO: 47)
- the L chain was prepared by changing the entire CDR of L404 to an amino acid excluding cysteine.
- the L chain on the anti-FX antibody side (heavy chain SEQ ID NO: 46, light chain SEQ ID NO: 47) is fixed to L404 of ACE910, and the L chain on the anti-FIX (a) antibody side contains all cysteines of L404 with cysteine. Substitutes modified with the amino acids except were prepared. Bispecific antibody expression and purification was performed by methods known to those skilled in the art.
- FVIII cofactor function substitution activity was evaluated by a method known to those skilled in the art using various purified bispecific antibodies. Specifically, it measured by the following method. All reactions were performed at room temperature. 5 ⁇ L of antibody solution diluted with Tris-buffered saline containing 0.1% bovine serum albumin (hereinafter referred to as TBSB) was mixed with 600 ng / mL Human Factor IXa beta (Enzyme Research Laboratories) 5 ⁇ L, and then in a 384-well plate. For 30 minutes at room temperature.
- TBSB Tris-buffered saline containing 0.1% bovine serum albumin
- the enzyme reaction in the mixture was started by adding 5 ⁇ L of 24.7 ⁇ g / mL Human Factor X X (Enzyme Research Laboratories), and 4 minutes later, it was stopped by adding 5 ⁇ L of 0.5 M EDTA.
- the color development reaction was initiated by adding 5 ⁇ L of color development substrate solution (S-2222, Sekisui® MEDICAL). After the color development reaction for 30 minutes, the absorbance change at 405 nm was measured using SpectraMax-340PC384- (Molecular Devices).
- Table 2 shows Kabat numbering (numbers in the “position” column at the left end), amino acids in ACE910 (amino acids listed in the second column from the left (single letter)), amino acids after mutation (most The specific activity for amino acid (single letter notation) and ACE910 (purified antibody) described in the upper row was calculated and corrected with the specific activity value of ACE910 (culture supernatant) (each numerical value in the table).
- Example 2 Acquisition of new L chain compatible with each H chain of ACE910 As shown in Example 1, FVIII complementation was performed by comprehensively adding amino acid modifications to L404, which is a common L chain of ACE910. The factor function alternative activity could be improved.
- a novel novel anti-FIX (a) antibody and anti-FX antibody H chain (Q499 and J327) has a completely different sequence from the common L chain. A method for obtaining the L chain from a human antibody library was considered.
- the present inventors refer to methods known to those skilled in the art, specifically, (Biochemical and Biophysical Research Communications, (2000), 275, 2, 553-557), etc.
- a new library replaced with a chain library is prepared, and then panning is performed on biotin-labeled human FIXa or biotin-labeled human FX to obtain an antibody with a novel L chain having FVIII cofactor function alternative activity.
- QNK131 (heavy chain SEQ ID NO: 45, light chain SEQ ID NO: 13), QNK284 (heavy chain sequence) as anti-human FIX
- JNK163 (heavy chain SEQ ID NO: 46, light chain SEQ ID NO: 20), JNK252 (heavy chain SEQ ID NO: 46, light chain SEQ ID NO: 21), JNK263 (heavy chain SEQ ID NO: 46, light chain) SEQ ID NO: 22), JNK339 (heavy chain SEQ ID NO: 46, light chain SEQ ID NO: 23), JNK348 (heavy chain SEQ ID NO: 46, light chain) Column number: 24), JNK351 (heavy chain sequence number: 46, light chain sequence number: 25), JNK360 (heavy chain sequence number: 46, light chain sequence number: 26), JNK378 (heavy chain sequence number: 46, light Chain sequence number: 27), JNK382 (heavy chain sequence number: 46, light chain sequence number: 28), JNL036 (heavy chain sequence number: 46, light chain sequence number: 29), JNL072 (heavy chain sequence number: 46, Light chain SEQ ID NO: 30), JNL095 (heavy chain SEQ ID NO:
- bispecific antibodies having these novel L chains were expressed and purified by methods known to those skilled in the art.
- the prepared antibodies are shown in Table 3 (clone name, heavy chain sequence number, anti-FIX (a) light chain sequence number or anti-FX light chain sequence number are shown). Note that the new L chain was used only for one side, and the other was L404 which is a common L chain of ACE910.
- the results of measuring the FVIII cofactor function-substituting activity of various purified bispecific antibodies are shown in FIG. It was confirmed that the bispecific antibody having any novel L chain has FVIII cofactor function substitution activity.
- Example 3 Preparation of a variant of a bispecific antibody having a novel L chain
- anti-FIX An amino acid substitution modification is introduced into QNK131 (light chain SEQ ID NO: 13), which is a novel L chain of an antibody, to obtain QAL187 (light chain SEQ ID NO: 42) and QAL201 (light chain SEQ ID NO: 43). did.
- amino acid substitution modification was introduced into JNL095 (light chain SEQ ID NO: 31), which is a novel L chain of anti-FX antibody, to obtain JYL280 (light chain SEQ ID NO: 44).
- a bispecific antibody (Q499 / QAL201 // J327 / JYL280 consisting of an anti-FIX (a) antibody consisting of heavy chain Q499 and light chain QAL201 and an anti-FX antibody consisting of heavy chain J327 and light chain JYL280 ) was expressed and purified by methods known to those skilled in the art, and the FVIII cofactor function alternative activity was measured and the results are shown in FIG.
- Anti-FIX (a) antibody-side heavy chain and light chain were fixed to Q499 and QAL201, anti-FX antibody side light chain was fixed to JYL280, and the J327 all CDRs were changed to amino acids excluding cysteine.
- Anti-FX antibody side heavy chain and light chain are fixed to J327 and JYL280 respectively, anti-FIX (a) antibody side light chain is fixed to QAL187, and all Q499 CDRs are replaced with all amino acids except cysteine
- Bispecific antibody expression and purification was performed by methods known to those skilled in the art.
- FVIII cofactor function alternative activity was evaluated by a method known to those skilled in the art using various purified bispecific antibodies.
- Table 4 shows FVIII cofactor function substitution activity against parent bispecific antibodies Q499 / QAL187 // J327 / JYL280 or Q499 / QAL201 // J327 / JYL280. It became clear that it could be improved.
- Table 4 shows Kabat numbering of mutation sites (numbers written in the “position” column at the left end), amino acids in the parent antibody (amino acids listed in the second column from the left (single letter code)), amino acids after mutation ( Specific activities for amino acids (single letter notation) listed in the top row and ACE910 (purified antibody) were shown (each numerical value in the table).
- (-) indicates that the expression level of the antibody was low, and "/" indicates that it was not produced because it was the same amino acid as itself.
- ACE910 (Emicizumab) has a light chain amino acid substitution that improves FVIII cofactor function substitution activity, a novel light chain that exhibits FVIII cofactor function substitution activity, and FVIII of a bispecific antibody having the novel light chain Heavy chain amino acid substitutions have been found to improve cofactor function alternative activity.
- amino acid substitutions and novel light chains are useful for the creation of bispecific antibodies having FVIII cofactor function substitution activity superior to ACE910 (Emicizumab).
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Abstract
Description
〔1〕抗体軽鎖可変ドメインを含むポリペプチドであって、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
〔2〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔3〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第一のポリペプチドは配列番号:45に記載の抗体重鎖可変ドメインのアミノ酸配列、第二のポリペプチドは配列番号:46に記載の抗体重鎖可変ドメインのアミノ酸配列をそれぞれ含み、第三のポリペプチド及び第四のポリペプチドのいずれか一方のポリペプチドは配列番号:47に記載の抗体軽鎖可変ドメインのアミノ酸配列を含み、他方のポリペプチドは、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、他方のポリペプチドはKabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔4〕抗体重鎖可変ドメインを含むポリペプチドであって、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
〔5〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔6〕抗体重鎖可変ドメインを含むポリペプチドであって、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、N52、T52a、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
〔7〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、N52、T52a、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔8〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第二のポリペプチドは配列番号:46に記載の抗体重鎖可変ドメインのアミノ酸配列、第三のポリペプチドは配列番号:42に記載の抗体軽鎖のアミノ酸配列、第四のポリペプチドは配列番号:44に記載の抗体軽鎖のアミノ酸配列をそれぞれ含み、第一のポリペプチドは、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、第一のポリペプチドはKabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔9〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第一のポリペプチドは配列番号:45に記載の抗体重鎖のアミノ酸配列、第三のポリペプチドは配列番号:43に記載の抗体軽鎖のアミノ酸配列、第四のポリペプチドは配列番号:44に記載の抗体軽鎖のアミノ酸配列をそれぞれ含み、第二のポリペプチドは、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、第二のポリペプチドはKabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
〔10〕抗体軽鎖であって、下記(a1)から(a6)、(b1)から(b23)及び(c1)から(c3)から選ばれるいずれかのアミノ酸配列を含む抗体軽鎖。
(a1) 配列番号:13に記載のアミノ酸配列を含む抗体軽鎖(QNK131)
(a2) 配列番号:14に記載のアミノ酸配列を含む抗体軽鎖(QNK284)
(a3) 配列番号:15に記載のアミノ酸配列を含む抗体軽鎖(QNK315)
(a4) 配列番号:16に記載のアミノ酸配列を含む抗体軽鎖(QNL182)
(a5) 配列番号:17に記載のアミノ酸配列を含む抗体軽鎖(QNL492)
(a6) 配列番号:18に記載のアミノ酸配列を含む抗体軽鎖(QNL576)
(b1) 配列番号:19に記載のアミノ酸配列を含む抗体軽鎖(JNK131)
(b2) 配列番号:20に記載のアミノ酸配列を含む抗体軽鎖(JNK163)
(b3) 配列番号:21に記載のアミノ酸配列を含む抗体軽鎖(JNK252)
(b4) 配列番号:22に記載のアミノ酸配列を含む抗体軽鎖(JNK263)
(b5) 配列番号:23に記載のアミノ酸配列を含む抗体軽鎖(JNK339)
(b6) 配列番号:24に記載のアミノ酸配列を含む抗体軽鎖(JNK348)
(b7) 配列番号:25に記載のアミノ酸配列を含む抗体軽鎖(JNK351)
(b8) 配列番号:26に記載のアミノ酸配列を含む抗体軽鎖(JNK360)
(b9) 配列番号:27に記載のアミノ酸配列を含む抗体軽鎖(JNK378)
(b10) 配列番号:28に記載のアミノ酸配列を含む抗体軽鎖(JNK382)
(b11) 配列番号:29に記載のアミノ酸配列を含む抗体軽鎖(JNL036)
(b12) 配列番号:30に記載のアミノ酸配列を含む抗体軽鎖(JNL072)
(b13) 配列番号:31に記載のアミノ酸配列を含む抗体軽鎖(JNL095)
(b14) 配列番号:32に記載のアミノ酸配列を含む抗体軽鎖(JNL176)
(b15) 配列番号:33に記載のアミノ酸配列を含む抗体軽鎖(JNL208)
(b16) 配列番号:34に記載のアミノ酸配列を含む抗体軽鎖(JNL224)
(b17) 配列番号:35に記載のアミノ酸配列を含む抗体軽鎖(JNL260)
(b18) 配列番号:36に記載のアミノ酸配列を含む抗体軽鎖(JNL056)
(b19) 配列番号:37に記載のアミノ酸配列を含む抗体軽鎖(JNL059)
(b20) 配列番号:38に記載のアミノ酸配列を含む抗体軽鎖(JNL226)
(b21) 配列番号:39に記載のアミノ酸配列を含む抗体軽鎖(JNL250)
(b22) 配列番号:40に記載のアミノ酸配列を含む抗体軽鎖(JNL263)
(b23) 配列番号:41に記載のアミノ酸配列を含む抗体軽鎖(JNL281)
(c1) 配列番号:42に記載のアミノ酸配列を含む抗体軽鎖(QAL187)
(c2) 配列番号:43に記載のアミノ酸配列を含む抗体軽鎖(QAL201)
(c3) 配列番号:44に記載のアミノ酸配列を含む抗体軽鎖(JYL280)
〔11〕FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチド(d)と第三のポリペプチド(f)が対を形成し、第二のポリペプチド(e)と第四のポリペプチド(g)が対を形成し、それぞれのポリペプチドが以下に記載のポリペプチドである抗体。
(d)第一のポリペプチドは(d1)又は(d2)である。
(d1)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメイン、あるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(d2)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(e)第二のポリペプチドは(e1)又は(e2)である。
(e1)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(e2)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f)第三のポリペプチドは(f1)、(f2)又は(f3)である。
(f1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(f2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f3)〔10〕に記載の(a1)から(a6)、および(c1)~(c2)のいずれかに記載のポリペプチド
(g)第四のポリペプチドは(g1)、(g2)又は(g3)である。
(g1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(g2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(g3)〔10〕に記載の(b1)から(b23)、および(c3)のいずれかに記載のポリペプチド。
〔12〕Emicizumab改変体を製造する方法であって、以下の工程(a)を含む、方法。
(a)以下の(i)~(iii)のうち一以上により置換する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換
〔13〕 Emicizumab改変体を単離する方法であって、以下の工程(a)~(c)を含む、方法。
(a)以下の(i)~(iii)のうち一以上によりEmicizumab改変体を産生する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換
(b)(a)において産生された該改変体のFVIII補因子機能代替活性を測定する工程;ならびに
(c)Emicizumabと比べてFVIII補因子機能代替活性が向上したEmicizumab改変体を選択する工程
を包含する、方法。
〔14〕抗体である、〔1〕、〔4〕および〔6〕のうちいずれか1つに記載のポリペプチド。
〔15〕ヒト化抗体、又はヒト抗体である、〔2〕、〔3〕、〔5〕、〔7〕~〔9〕、〔11〕および〔14〕のうちいずれか1つに記載の抗体。
〔16〕Fv、Fab、Fab'、Fab'-SH、F(ab')2、ダイアボディ、線状抗体、単鎖抗体分子および抗体断片から形成された多重特異性抗体からなる群から選択される抗体である、〔2〕、〔3〕、〔5〕、〔7〕~〔9〕、〔11〕、〔14〕および〔15〕のいずれか1つに記載の抗体。
〔17〕〔2〕、〔3〕、〔5〕、〔7〕~〔9〕、〔11〕および〔14〕~〔16〕のいずれか1つに記載の抗体をコードする核酸。
〔18〕〔17〕に記載の核酸が挿入されたベクター。
〔19〕〔17〕に記載の核酸または〔17〕に記載のベクターを含む細胞。
〔20〕〔2〕、〔3〕、〔5〕、〔7〕~〔9〕、〔11〕および〔14〕~〔16〕のいずれかに記載の抗体ならびに薬学的に許容される担体を含む、薬学的製剤。
〔21〕出血、出血を伴う疾患、もしくは出血に起因する疾患の予防および/または治療に用いられる薬学的製剤であって、該疾患が、血液凝固第VIII因子および/または活性化血液凝固第VIII因子の活性の低下ないし欠損によって発症および/または進展する疾患である、〔20〕に記載の薬学的製剤。
〔22〕血液凝固第VIII因子および/または活性化血液凝固第VIII因子の活性の低下ないし欠損によって発症および/または進展する疾患が、血友病Aである、〔21〕に記載の薬学的製剤。
〔23〕血液凝固第VIII因子および/または活性化血液凝固第VIII因子の活性の低下ないし欠損によって発症および/または進展する疾患が、血液凝固第VIII因子および/または活性化血液凝固第VIII因子に対するインヒビターが出現している疾患である、〔22〕に記載の薬学的製剤。
〔24〕血液凝固第VIII因子および/または活性化血液凝固第VIII因子の活性の低下ないし欠損によって発症および/または進展する疾患が、後天性血友病である、〔21〕に記載の薬学的製剤。
〔25〕血液凝固第VIII因子および/または活性化血液凝固第VIII因子の活性の低下ないし欠損によって発症および/または進展する疾患が、フォンビルブランド病である、〔21〕に記載の薬学的製剤。
〔26〕少なくとも〔2〕、〔3〕、〔5〕、〔7〕~〔9〕、〔11〕および〔14〕~〔16〕のいずれか1つに記載の抗体を含む、出血、出血を伴う疾患もしくは出血に起因する疾患を予防および/または治療する方法に用いるための治療用品の商用パッケージ。
(a) アミノ酸残基26-32 (L1)、50-52 (L2)、91-96 (L3)、26-32 (H1)、53-55 (H2)、および96-101 (H3)のところで生じる超可変ループ (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987));
(b) アミノ酸残基24-34 (L1)、50-56 (L2)、89-97 (L3)、31-35b (H1)、50-65 (H2)、 および95-102 (H3)のところで生じるCDR (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991));
(c) アミノ酸残基27c-36 (L1)、46-55 (L2)、89-96 (L3)、30-35b (H1)、47-58 (H2)、および93-101 (H3) のところで生じる抗原接触 (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996));ならびに、
(d) HVRアミノ酸残基46-56 (L2)、47-56 (L2)、48-56 (L2)、49-56 (L2)、26-35 (H1)、26-35b (H1)、49-65 (H2)、93-102 (H3)、および94-102 (H3)を含む、(a)、(b)、および/または(c)の組合せ。
別段示さない限り、HVR残基および可変ドメイン中の他の残基(例えば、FR残基)は、本明細書では上記のKabatらにしたがって番号付けされる。
さらに具体的には、前記二重特異性抗体は、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成する二重特異性抗体であって、第一のポリペプチドが配列番号:45に記載の抗体重鎖可変ドメインのアミノ酸配列を含むH鎖、第二のポリペプチドが配列番号:46に記載の抗体重鎖可変ドメインのアミノ酸配列を含むH鎖、および第三のポリペプチドと第四のポリペプチドが配列番号:47に記載の抗体軽鎖可変ドメインのアミノ酸配列を含む共通L鎖からなる二重特異性抗体(Q499-z121/J327-z119/L404-k)である。
さらに具体的には、前記二重特異性抗体は、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成する二重特異性抗体であって、第一のポリペプチドが配列番号:10に記載のアミノ酸配列を含むH鎖、第二のポリペプチドが配列番号:11に記載のアミノ酸配列を含むH鎖、および第三のポリペプチドと第四のポリペプチドが配列番号:12に記載のアミノ酸配列を含む共通L鎖からなる二重特異性抗体(Q499-z121/J327-z119/L404-k)である。
このような抗体は、例えばWO2005/035756、WO2006/109592、WO2012/067176などに記載の方法に従って取得することができる。以下、本明細書ではEmicizumabとACE910は同義である。
例えばEmicizumab改変体とは以下にあげるポリペプチド及び抗体であるが、これらに限定されるものではない。
(a1) 配列番号:13に記載のアミノ酸配列を含む抗体軽鎖(QNK131)
(a2) 配列番号:14に記載のアミノ酸配列を含む抗体軽鎖(QNK284)
(a3) 配列番号:15に記載のアミノ酸配列を含む抗体軽鎖(QNK315)
(a4) 配列番号:16に記載のアミノ酸配列を含む抗体軽鎖(QNL182)
(a5) 配列番号:17に記載のアミノ酸配列を含む抗体軽鎖(QNL492)
(a6) 配列番号:18に記載のアミノ酸配列を含む抗体軽鎖(QNL576)
(b1) 配列番号:19に記載のアミノ酸配列を含む抗体軽鎖(JNK131)
(b2) 配列番号:20に記載のアミノ酸配列を含む抗体軽鎖(JNK163)
(b3) 配列番号:21に記載のアミノ酸配列を含む抗体軽鎖(JNK252)
(b4) 配列番号:22に記載のアミノ酸配列を含む抗体軽鎖(JNK263)
(b5) 配列番号:23に記載のアミノ酸配列を含む抗体軽鎖(JNK339)
(b6) 配列番号:24に記載のアミノ酸配列を含む抗体軽鎖(JNK348)
(b7) 配列番号:25に記載のアミノ酸配列を含む抗体軽鎖(JNK351)
(b8) 配列番号:26に記載のアミノ酸配列を含む抗体軽鎖(JNK360)
(b9) 配列番号:27に記載のアミノ酸配列を含む抗体軽鎖(JNK378)
(b10) 配列番号:28に記載のアミノ酸配列を含む抗体軽鎖(JNK382)
(b11) 配列番号:29に記載のアミノ酸配列を含む抗体軽鎖(JNL036)
(b12) 配列番号:30に記載のアミノ酸配列を含む抗体軽鎖(JNL072)
(b13) 配列番号:31に記載のアミノ酸配列を含む抗体軽鎖(JNL095)
(b14) 配列番号:32に記載のアミノ酸配列を含む抗体軽鎖(JNL176)
(b15) 配列番号:33に記載のアミノ酸配列を含む抗体軽鎖(JNL208)
(b16) 配列番号:34に記載のアミノ酸配列を含む抗体軽鎖(JNL224)
(b17) 配列番号:35に記載のアミノ酸配列を含む抗体軽鎖(JNL260)
(b18) 配列番号:36に記載のアミノ酸配列を含む抗体軽鎖(JNL056)
(b19) 配列番号:37に記載のアミノ酸配列を含む抗体軽鎖(JNL059)
(b20) 配列番号:38に記載のアミノ酸配列を含む抗体軽鎖(JNL226)
(b21) 配列番号:39に記載のアミノ酸配列を含む抗体軽鎖(JNL250)
(b22) 配列番号:40に記載のアミノ酸配列を含む抗体軽鎖(JNL263)
(b23) 配列番号:41に記載のアミノ酸配列を含む抗体軽鎖(JNL281)
特定の態様において、本発明は、上記(a1)から(a6)及び(b1)から(b23)の抗体L鎖の変異体を提供する。
(c1) 配列番号:13に記載の抗体軽鎖の変異体であって配列番号:42に記載のアミノ酸配列を有する抗体軽鎖(QAL187)
(c2) 配列番号:13に記載の抗体軽鎖の変異体であって配列番号:43に記載のアミノ酸配列を有する抗体L鎖(QAL201)
(c3) 配列番号:31に記載の抗体軽鎖の変異体であって配列番号:44に記載のアミノ酸配列を有する抗体L鎖(JYL280)
(d)第一のポリペプチドは(d1)又は(d2)である。
(d1)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメイン、あるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(d2)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(e)第二のポリペプチドは(e1)又は(e2)である。
(e1)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメイン、あるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(e2)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f)第三のポリペプチドは(f1)、(f2)又は(f3)である。
(f1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(f2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f3)〔11〕に記載の(a1)から(a6)、および(c1)~(c2)のいずれかに記載のポリペプチド
(g)第四のポリペプチドは(g1)、(g2)又は(g3)である。
(g1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(g2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(g3)〔11〕に記載の(b1)から(b23)、および(c3)のいずれかに記載のポリペプチド。
前記ポリペプチドあるいは抗体において、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインと異なるアミノ酸残基、あるいは配列番号:47に記載の抗体軽鎖可変ドメインと異なるアミノ酸残基の数は一つ以上であればよく、好ましくは2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20であり、さらに好ましくは21以上である。尚、置換位置はKabat番号付けシステムに従って番号付けられる位置を示す。
K24:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、R、F、W、Y
A25:I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
S26:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
R27:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
N28:A、I、L、M、P、V、G、Q、S、T、D、E、H、K、R、F、W、Y
I29:A、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
E30:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
R31:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
Q32:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
L33:A、I、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
A34:I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
Q50:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
A51:I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
S52:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
R53:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
K54:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、R、F、W、Y
E55:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
S56:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
Q89:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
Q90:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
Y91:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
S92:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
D93:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
P94:A、I、L、M、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
P95:A、I、L、M、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
L96:A、I、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
T97:A、I、L、M、P、V、G、N、Q、S、D、E、H、K、R、F、W、Y
前記ポリペプチドあるいは抗体において、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインと異なるアミノ酸残基、あるいは配列番号:45に記載の抗体重鎖可変ドメインと異なるアミノ酸残基の数は一つ以上であればよく、好ましくは2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20であり、さらに好ましくは21以上である。尚、置換位置はKabat番号付けシステムに従って番号付けられる位置を示す。
Y31:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
Y32:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
D33:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
I34:A、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
Q35:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
S50:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
I51:A、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
S52:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
P52a:A、I、L、M、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
S53:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
G54:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
Q55:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
S56:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
T57:A、I、L、M、P、V、G、N、Q、S、D、E、H、K、R、F、W、Y
Y58:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
Y59:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
R60:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
R61:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
E62:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
V63:A、I、L、M、P、G、N、Q、S、T、D、E、H、K、R、F、W、Y
K64:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、R、F、W、Y
G65:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
R95:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
T96:A、I、L、M、P、V、G、N、Q、S、D、E、H、K、R、F、W、Y
G97:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
R98:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
E99:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
Y100:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
G100a:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
G100b:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
G100c:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
W100d:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、Y
Y100e:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
F100f:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、W、Y
D101:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
Y102:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
前記ポリペプチドあるいは抗体において、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインと異なるアミノ酸残基、あるいは配列番号:46に記載の抗体重鎖可変ドメインと異なるアミノ酸残基の数は一つ以上であればよく、好ましくは2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20であり、さらに好ましくは21以上である。尚、置換位置はKabat番号付けシステムに従って番号付けられる位置を示す。
D31:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
N32:A、I、L、M、P、V、G、Q、S、T、D、E、H、K、R、F、W、Y
N33:A、I、L、M、P、V、G、Q、S、T、D、E、H、K、R、F、W、Y
M34:A、I、L、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
D35:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
D50:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
I51:A、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
N52:A、I、L、M、P、V、G、Q、S、T、D、E、H、K、R、F、W、Y
T52a:A、I、L、M、P、V、G、N、Q、S、D、E、H、K、R、F、W、Y
R53:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
S54:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
G55:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
G56:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
S57:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
I58:A、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
Y59:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
N60:A、I、L、M、P、V、G、Q、S、T、D、E、H、K、R、F、W、Y
E61:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
E62:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
F63:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、W、Y
Q64:A、I、L、M、P、V、G、N、S、T、D、E、H、K、R、F、W、Y
D65:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
R95:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、F、W、Y
K96:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、R、F、W、Y
S97:A、I、L、M、P、V、G、N、Q、T、D、E、H、K、R、F、W、Y
Y98:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
G99:A、I、L、M、P、V、N、Q、S、T、D、E、H、K、R、F、W、Y
Y100:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
Y100a:A、I、L、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W
L100b:A、I、M、P、V、G、N、Q、S、T、D、E、H、K、R、F、W、Y
D101:A、I、L、M、P、V、G、N、Q、S、T、E、H、K、R、F、W、Y
E102:A、I、L、M、P、V、G、N、Q、S、T、D、H、K、R、F、W、Y
特定の態様において、本明細書で提供される変異体には、アミノ酸配列中のアミノ酸残基の欠失、および/またはアミノ酸配列中へのアミノ酸残基の挿入、および/またはアミノ酸配列中のアミノ酸残基の置換が行われた変異体が含まれる。最終構築物が向上したFVIII補因子機能代替活性を備えることを前提に、欠失、挿入、および置換の任意の組合せが、最終構築物に至るために行われ得る。
以下に本明細書で提供されるFIXおよび/またはFIXaならびにFXを認識する二重特異性抗体(以下、単に「抗FIX(a)/FX二重特異性抗体」という)の例を記載するが、以下は本明細書で提供される他の抗体及びポリペプチドにも同様に当てはまる。尚、本明細書において、FIX(a)はFIXおよび/またはFIXaを意味する。
特定の態様において、本明細書で提供される抗体は、抗体断片である。抗体断片は、これらに限定されるものではないが、Fab、Fab'、Fab'-SH、F(ab')2、Fv、および scFv断片、ならびに、後述する他の断片を含む。特定の抗体断片についての総説として、Hudson et al. Nat. Med. 9:129-134 (2003) を参照のこと。scFv断片の総説として、例えば、Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp.269-315 (1994);加えて、WO93/16185;ならびに米国特許第5,571,894号および第5,587,458号を参照のこと。サルベージ受容体結合エピトープ残基を含みインビボ (in vivo) における半減期の長くなったFabおよびF(ab')2断片についての論説として、米国特許第5,869,046号を参照のこと。
特定の態様において、本明細書で提供される抗体は、ヒト抗体である。ヒト抗体は、当該技術分野において知られる種々の手法によって製造され得る。ヒト抗体は、van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) および Lonberg, Curr. Opin. Immunol. 20:450-459 (2008) に、概説されている。
本発明の抗体は、所望の1つまたは複数の活性を伴う抗体についてコンビナトリアルライブラリをスクリーニングすることによって単離してもよい。例えば、ファージディスプレイライブラリの生成や、所望の結合特性を備える抗体についてそのようなライブラリをスクリーニングするための、様々な方法が当該技術分野において知られている。そのような方法は、Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001) において総説されており、さらに例えば、McCafferty et al., Nature 348:552-554;Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004);およびLee et al., J. Immunol. Methods 284(1-2): 119-132(2004) に記載されている。
特定の態様において、1つまたは複数のアミノ酸置換を有する抗体変異体が提供される。置換的変異導入の目的部位は、HVRおよびFRを含む。保存的置換を、表1の「好ましい置換」の見出しの下に示す。より実質的な変更を、表1の「例示的な置換」の見出しの下に提供するとともに、アミノ酸側鎖のクラスに言及しつつ下で詳述する。アミノ酸置換は目的の抗体に導入されてもよく、産物は、例えば、保持/改善された抗原結合性、減少した免疫原性、または改善したFVIII補因子機能代替活性、ADCCまたはCDCなどの、所望の活性についてスクリーニングされてもよい。
(1) 疎水性:ノルロイシン、メチオニン (Met)、アラニン (Ala)、バリン (Val)、ロイシン (Leu)、イソロイシン (Ile);
(2) 中性の親水性:システイン (Cys)、セリン (Ser)、トレオニン (Thr)、アスパラギン (Asn)、グルタミン (Gln);
(3) 酸性:アスパラギン酸 (Asp)、グルタミン酸 (Glu);
(4) 塩基性:ヒスチジン (His)、リジン (Lys)、アルギニン (Arg);
(5) 鎖配向に影響する残基:グリシン (Gly)、プロリン (Pro);
(6) 芳香族性:トリプトファン (Trp)、チロシン (Tyr)、フェニルアラニン (Phe)。
非保存的置換は、これらのクラスの1つのメンバーを、別のクラスのものに交換することをいう。
特定の態様において、本明細書で提供される抗体は、抗体がグリコシル化される程度を増加させるまたは減少させるように改変されている。抗体へのグリコシル化部位の追加または削除は、1つまたは複数のグリコシル化部位を作り出すまたは取り除くようにアミノ酸配列を改変することにより、簡便に達成可能である。
特定の態様において、本明細書で提供される抗体のFc領域に1つまたは複数のアミノ酸修飾を導入して、それによりFc領域変異体を生成してもよい。Fc領域変異体は、1つまたは複数のアミノ酸ポジションのところでアミノ酸修飾(例えば、置換)を含む、ヒトFc領域配列(例えば、ヒトIgG1、IgG2、IgG3、またはIgG4のFc領域)を含んでもよい。
a) N434A/Y436T/Q438R/S440E
b) N434A/Y436V/Q438R/S440E
c) M428L/N434A/Y436T/Q438R/S440E
d) M428L/N434A/Y436V/Q438R/S440E
e) M428L/N434A/Q438R/S440E
f) N434A/Q438R/S440E
特定の態様において、本明細書で提供される抗体は、当該技術分野において知られておりかつ容易に入手可能な追加の非タンパク質部分を含むように、さらに修飾されてもよい。抗体の誘導体化に好適な部分は、これに限定されるものではないが、水溶性ポリマーを含む。水溶性ポリマーの非限定的な例は、これらに限定されるものではないが、ポリエチレングリコール (PEG)、エチレングリコール/プロピレングリコールのコポリマー、カルボキシメチルセルロース、デキストラン、ポリビニルアルコール、ポリビニルピロリドン、ポリ1,3ジオキソラン、ポリ1,3,6,トリオキサン、エチレン/無水マレイン酸コポリマー、ポリアミノ酸(ホモポリマーまたはランダムコポリマーのいずれでも)、および、デキストランまたはポリ(n-ビニルピロリドン)ポリエチレングリコール、ポリプロピレングリコールホモポリマー、ポリプロピレンオキシド/エチレンオキシドコポリマー、ポリオキシエチル化ポリオール類(例えばグリセロール)、ポリビニルアルコール、および、これらの混合物を含む。ポリエチレングリコールプロピオンアルデヒドは、その水に対する安定性のために、製造において有利であるだろう。ポリマーは、いかなる分子量でもよく、枝分かれしていてもしていなくてもよい。抗体に付加されるポリマーの数には幅があってよく、1つ以上のポリマーが付加されるならそれらは同じ分子であってもよいし、異なる分子であってもよい。一般的に、誘導体化に使用されるポリマーの数および/またはタイプは、これらに限定されるものではないが、改善されるべき抗体の特定の特性または機能、抗体誘導体が規定の条件下での療法に使用されるか否か、などへの考慮に基づいて、決定することができる。
例えば、米国特許第4,816,567号に記載されるとおり、抗体は組み換えの方法や構成を用いて製造することができる。一態様において、本明細書に記載の抗FIX(a)/FX二重特異性抗体をコードする、単離された核酸が提供される。そのような核酸は、抗体のVLを含むアミノ酸配列および/またはVHを含むアミノ酸配列(例えば、抗体の軽鎖および/または重鎖)をコードしてもよい。さらなる態様において、このような核酸を含む1つまたは複数のベクター(例えば、発現ベクター)が提供される。さらなる態様において、このような核酸を含む宿主細胞が提供される。このような態様の1つでは、宿主細胞は、(1)抗体のVLを含むアミノ酸配列および抗体のVHを含むアミノ酸配列をコードする核酸を含むベクター、または、(2)抗体のVLを含むアミノ酸配列をコードする核酸を含む第一のベクターと抗体のVHを含むアミノ酸配列をコードする核酸を含む第二のベクターを含む(例えば、形質転換されている)。一態様において、宿主細胞は、真核性である(例えば、チャイニーズハムスター卵巣 (CHO) 細胞)またはリンパ系の細胞(例えば、Y0、NS0、Sp2/0細胞))。一態様において、抗FIX(a)/FX二重特異性抗体の発現に好適な条件下で、上述のとおり当該抗体をコードする核酸を含む宿主細胞を培養すること、および任意で、当該抗体を宿主細胞(または宿主細胞培養培地)から回収することを含む、抗FIX(a)/FX二重特異性抗体を作製する方法が提供される。
本明細書で提供される抗FIX(a)/FX二重特異性抗体は、当該技術分野において知られている種々の測定法によって、同定され、スクリーニングされ、または物理的/化学的特性および/または生物学的活性について明らかにされてもよい。
一局面において、本発明の抗体は、例えばELISA、ウエスタンブロット等の公知の方法によって、その抗原結合活性に関して試験される。
一局面において、生物学的活性を有する抗FIX(a)/FX二重特異性抗体のそれを同定するための測定法が提供される。生物学的活性は、例えば、FVIII補因子機能代替活性を含んでよい。また、このような生物学的活性をインビボおよび/またはインビトロで有する抗体が、提供される。
本明細書に記載の抗FIX(a)/FX二重特異性抗体の薬学的製剤は、所望の純度を有する抗体を、1つまたは複数の任意の薬学的に許容される担体 (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) と混合することによって、凍結乾燥製剤または水溶液の形態で、調製される。薬学的に許容される担体は、概して、用いられる際の用量および濃度ではレシピエントに対して非毒性であり、これらに限定されるものではないが、以下のものを含む:リン酸塩、クエン酸塩、および他の有機酸などの緩衝液;アスコルビン酸およびメチオニンを含む、抗酸化剤;保存料(オクタデシルジメチルベンジル塩化アンモニウム;塩化ヘキサメトニウム;塩化ベンザルコニウム;塩化ベンゼトニウム;フェノール、ブチル、またはベンジルアルコール;メチルまたはプロピルパラベンなどのアルキルパラベン;カテコール;レソルシノール;シクロヘキサノール;3-ペンタノール;およびm-クレゾールなど);低分子(約10残基未満)ポリペプチド;血清アルブミン、ゼラチン、または免疫グロブリンなどのタンパク質;ポリビニルピロリドンなどの親水性ポリマー;グリシン、グルタミン、アスパラギン、ヒスチジン、アルギニン、またはリジンなどのアミノ酸;グルコース、マンノース、またはデキストリンを含む、単糖、二糖、および他の炭水化物;EDTAなどのキレート剤;スクロース、マンニトール、トレハロース、ソルビトールなどの、砂糖類;ナトリウムなどの塩形成対イオン類;金属錯体(例えば、Zn-タンパク質錯体);および/またはポリエチレングリコール (PEG) などの非イオン系表面活性剤。本明細書の例示的な薬学的に許容される担体は、さらに、可溶性中性活性型ヒアルロニダーゼ糖タンパク質 (sHASEGP)(例えば、rHuPH20 (HYLENEX(登録商標)、Baxter International, Inc.) などのヒト可溶性PH-20ヒアルロニダーゼ糖タンパク質)などの間質性薬剤分散剤を含む。特定の例示的sHASEGPおよびその使用方法は(rHuPH20を含む)、米国特許出願公開第2005/0260186号および第2006/0104968号に記載されている。一局面において、sHASEGPは、コンドロイチナーゼなどの1つまたは複数の追加的なグリコサミノグリカナーゼと組み合わせられる。
本明細書で提供される抗FIX(a)/FX二重特異性抗体のいずれも、治療的な方法において使用されてよい。
一局面において、医薬品としての使用のための、抗FIX(a)/FX二重特異性抗体が提供される。さらなる局面において、出血、出血を伴う疾患、もしくは出血に起因する疾患の治療における使用のための、抗FIX(a)/FX二重特異性抗体が提供される。特定の態様において、治療方法における使用のための、抗FIX(a)/FX二重特異性抗体が提供される。特定の態様において、本発明は、出血、出血を伴う疾患、もしくは出血に起因する疾患を有する個体を治療する方法であって、当該個体に抗FIX(a)/FX二重特異性抗体の有効量を投与する工程を含む方法における使用のための、抗FIX(a)/FX二重特異性抗体を提供する。このような態様の1つにおいて、方法は、当該個体に少なくとも1つの(例えば後述するような)追加治療剤の有効量を投与する工程を、さらに含む。さらなる態様において、本発明は、FVIII機能の代替における使用のための抗FIX(a)/FX二重特異性抗体を提供する。特定の態様において、本発明は、個体においてFVIIIの機能を代替する方法であって、FVIIIの機能を代替するために当該個体に抗FIX(a)/FX二重特異性抗体の有効量を投与する工程を含む方法における使用のための、抗FIX(a)/FX二重特異性抗体を提供する。上記態様の任意のものによる「個体」は、好適にはヒトである。
本発明の別の局面において、上述の障害の治療、予防、および/または診断に有用な器材を含んだ製品が、提供される。製品は、容器、および当該容器上のラベルまたは当該容器に付属する添付文書を含む。好ましい容器としては、例えば、ボトル、バイアル、シリンジ、IV溶液バッグなどが含まれる。容器類は、ガラスやプラスチックなどの、様々な材料から形成されていてよい。容器は組成物を単体で保持してもよいし、症状の治療、予防、および/または診断のために有効な別の組成物と組み合わせて保持してもよく、また、無菌的なアクセスポートを有していてもよい(例えば、容器は、皮下注射針によって突き通すことのできるストッパーを有する静脈内投与用溶液バッグまたはバイアルであってよい)。組成物中の少なくとも1つの有効成分は、本発明の抗体である。ラベルまたは添付文書は、組成物が選ばれた症状を治療するために使用されるものであることを示す。さらに製品は、(a)第一の容器であって、その中に収められた本発明の抗体を含む組成物を伴う、第一の容器;および、(b)第二の容器であって、その中に収められたさらなる細胞傷害剤またはそれ以外で治療的な剤を含む組成物を伴う、第二の容器を含んでもよい。本発明のこの態様における製品は、さらに、組成物が特定の症状を治療するために使用され得ることを示す、添付文書を含んでもよい。あるいはまたは加えて、製品はさらに、注射用制菌水 (BWFI)、リン酸緩衝生理食塩水、リンガー溶液、およびデキストロース溶液などの、薬学的に許容される緩衝液を含む、第二の(または第三の)容器を含んでもよい。他の緩衝液、希釈剤、フィルター、針、およびシリンジなどの、他の商業的観点またはユーザの立場から望ましい器材をさらに含んでもよい。
(a)以下の(i)~(iii)のうち一以上により置換する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換。
上記置換する個所は1以上であればよく、また(i)と(ii)の組み合わせ、(i)と(iii)の組み合わせ、(ii)と(iii)の組み合わせ、(i)と(ii)と(iii)の組み合わせであってもよい。
(a)以下の(i)~(iii)のうち一以上によりEmicizumab改変体を産生する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換
(b)(a)において産生された該改変抗体のFVIII補因子機能代替活性を測定する工程;ならびに
(c)Emicizumabと比べてFVIII補因子機能代替活性が向上したEmicizumab改変抗体を選択する工程。
上記置換する個所は1以上であればよく、また(i)と(ii)の組み合わせ、(i)と(iii)の組み合わせ、(ii)と(iii)の組み合わせ、(i)と(ii)と(iii)の組み合わせであってもよい。
ACE910は、FVIII補因子機能代替活性を示す抗FIX(a)および抗FXからなるヒト化IgG4抗体であり、FIX(a)およびFXそれぞれを認識する2種類の重鎖(Q499およびJ327)と共通L鎖(L404)から構成される(重鎖配列番号:45および46、軽鎖配列番号:47)。本発明者らは、変異導入のためのPCR等の当業者公知の方法により、L404に対してアミノ酸改変を網羅的に導入し、大規模にFVIII補因子機能代替活性のスクリーニングを実施することで、ACE910のFVIII補因子機能代替活性を向上させるL鎖のアミノ酸置換を見出した。
実施例1で示したように、ACE910の共通L鎖であるL404に対してアミノ酸改変を網羅的に加えることにより、FVIII補因子機能代替活性を向上させることができた。一方、ACE910のFVIII補因子機能代替活性を向上させる方法として、抗FIX(a)抗体および抗FX抗体のそれぞれのH鎖(Q499およびJ327)に対して共通L鎖とは全く異なる配列を有する新規L鎖をヒト抗体ライブラリから取得する方法が考えられた。
実施例2において取得した新規L鎖を有する二重特異性抗体のFVIII補因子機能代替活性を向上させるために、抗FIX(a)抗体の新規L鎖であるQNK131(軽鎖配列番号:13)に対してアミノ酸置換改変を導入し、QAL187(軽鎖配列番号:42)、QAL201(軽鎖配列番号:43)を取得した。同様に、抗FX抗体の新規L鎖であるJNL095(軽鎖配列番号:31)に対してアミノ酸置換改変を導入し、JYL280(軽鎖配列番号:44)を取得した。一例として、重鎖Q499と軽鎖QAL201からなる抗FIX(a)抗体および、重鎖J327と軽鎖JYL280からなる抗FX抗体から構成される二重特異性抗体(Q499/QAL201//J327/JYL280)を当業者公知の方法で発現、精製し、FVIII補因子機能代替活性を測定した結果を図2に示す。
FVIII補因子機能代替活性が向上した新規L鎖(抗FIX(a)抗体はQAL187、抗FX抗体はJYL280)を用い、Q499およびJ327に対して、アミノ酸変異を網羅的に導入し、大規模にFVIII補因子機能代替活性のスクリーニングを実施することで、FVIII補因子機能代替活性を向上させるアミノ酸置換を見出した。
精製された各種二重特異性抗体を用いて当業者公知の方法によりFVIII補因子機能代替活性を評価した。
Claims (13)
- 抗体軽鎖可変ドメインを含むポリペプチドであって、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第一のポリペプチドは配列番号:45に記載の抗体重鎖可変ドメインのアミノ酸配列、第二のポリペプチドは配列番号:46に記載の抗体重鎖可変ドメインのアミノ酸配列をそれぞれ含み、第三のポリペプチド及び第四のポリペプチドのいずれか一方のポリペプチドは配列番号:47に記載の抗体軽鎖可変ドメインのアミノ酸配列を含み、他方のポリペプチドは、配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含み、ここで、他方のポリペプチドはKabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- 抗体重鎖可変ドメインを含むポリペプチドであって、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- 抗体重鎖可変ドメインを含むポリペプチドであって、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、N52、T52a、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、N52、T52a、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第二のポリペプチドは配列番号:46に記載の抗体重鎖可変ドメインのアミノ酸配列、第三のポリペプチドは配列番号:42に記載の抗体軽鎖のアミノ酸配列、第四のポリペプチドは配列番号:44に記載の抗体軽鎖のアミノ酸配列をそれぞれ含み、第一のポリペプチドは、配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、第一のポリペプチドはKabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチドと第三のポリペプチドが対を形成し、第二のポリペプチドと第四のポリペプチドが対を形成し、第一のポリペプチドは配列番号:45に記載の抗体重鎖のアミノ酸配列、第三のポリペプチドは配列番号:43に記載の抗体軽鎖のアミノ酸配列、第四のポリペプチドは配列番号:44に記載の抗体軽鎖のアミノ酸配列をそれぞれ含み、第二のポリペプチドは、配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含み、ここで、第二のポリペプチドはKabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、抗体。
- 抗体軽鎖であって、下記(a1)から(a6)、(b1)から(b23)及び(c1)から(c3)から選ばれるいずれかのアミノ酸配列を含む抗体軽鎖。
(a1) 配列番号:13に記載のアミノ酸配列を含む抗体軽鎖(QNK131)
(a2) 配列番号:14に記載のアミノ酸配列を含む抗体軽鎖(QNK284)
(a3) 配列番号:15に記載のアミノ酸配列を含む抗体軽鎖(QNK315)
(a4) 配列番号:16に記載のアミノ酸配列を含む抗体軽鎖(QNL182)
(a5) 配列番号:17に記載のアミノ酸配列を含む抗体軽鎖(QNL492)
(a6) 配列番号:18に記載のアミノ酸配列を含む抗体軽鎖(QNL576)
(b1) 配列番号:19に記載のアミノ酸配列を含む抗体軽鎖(JNK131)
(b2) 配列番号:20に記載のアミノ酸配列を含む抗体軽鎖(JNK163)
(b3) 配列番号:21に記載のアミノ酸配列を含む抗体軽鎖(JNK252)
(b4) 配列番号:22に記載のアミノ酸配列を含む抗体軽鎖(JNK263)
(b5) 配列番号:23に記載のアミノ酸配列を含む抗体軽鎖(JNK339)
(b6) 配列番号:24に記載のアミノ酸配列を含む抗体軽鎖(JNK348)
(b7) 配列番号:25に記載のアミノ酸配列を含む抗体軽鎖(JNK351)
(b8) 配列番号:26に記載のアミノ酸配列を含む抗体軽鎖(JNK360)
(b9) 配列番号:27に記載のアミノ酸配列を含む抗体軽鎖(JNK378)
(b10) 配列番号:28に記載のアミノ酸配列を含む抗体軽鎖(JNK382)
(b11) 配列番号:29に記載のアミノ酸配列を含む抗体軽鎖(JNL036)
(b12) 配列番号:30に記載のアミノ酸配列を含む抗体軽鎖(JNL072)
(b13) 配列番号:31に記載のアミノ酸配列を含む抗体軽鎖(JNL095)
(b14) 配列番号:32に記載のアミノ酸配列を含む抗体軽鎖(JNL176)
(b15) 配列番号:33に記載のアミノ酸配列を含む抗体軽鎖(JNL208)
(b16) 配列番号:34に記載のアミノ酸配列を含む抗体軽鎖(JNL224)
(b17) 配列番号:35に記載のアミノ酸配列を含む抗体軽鎖(JNL260)
(b18) 配列番号:36に記載のアミノ酸配列を含む抗体軽鎖(JNL056)
(b19) 配列番号:37に記載のアミノ酸配列を含む抗体軽鎖(JNL059)
(b20) 配列番号:38に記載のアミノ酸配列を含む抗体軽鎖(JNL226)
(b21) 配列番号:39に記載のアミノ酸配列を含む抗体軽鎖(JNL250)
(b22) 配列番号:40に記載のアミノ酸配列を含む抗体軽鎖(JNL263)
(b23) 配列番号:41に記載のアミノ酸配列を含む抗体軽鎖(JNL281)
(c1) 配列番号:42に記載のアミノ酸配列を含む抗体軽鎖(QAL187)
(c2) 配列番号:43に記載のアミノ酸配列を含む抗体軽鎖(QAL201)
(c3) 配列番号:44に記載のアミノ酸配列を含む抗体軽鎖(JYL280) - FIXおよび/またはFIXaならびにFXを認識する二重特異性抗体であって、第一のポリペプチド(d)と第三のポリペプチド(f)が対を形成し、第二のポリペプチド(e)と第四のポリペプチド(g)が対を形成し、それぞれのポリペプチドが以下に記載のポリペプチドである抗体。
(d)第一のポリペプチドは(d1)又は(d2)である。
(d1)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメイン、あるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(d2)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:45に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(e)第二のポリペプチドは(e1)又は(e2)である。
(e1)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチド
(e2)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を有する抗体重鎖可変ドメインあるいは配列番号:46に記載のアミノ酸配列を有する抗体重鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f)第三のポリペプチドは(f1)、(f2)又は(f3)である。
(f1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(f2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(f3)請求項10に記載の(a1)から(a6)、および(c1)~(c2)のいずれかに記載のポリペプチド
(g)第四のポリペプチドは(g1)、(g2)又は(g3)である。
(g1)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメイン、あるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチド
(g2)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を有する抗体軽鎖可変ドメインあるいは配列番号:47に記載のアミノ酸配列を有する抗体軽鎖可変ドメインを含むポリペプチドであって、Kabat番号付けシステムに従って番号付けられた、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸が、システイン以外の任意のアミノ酸に置換されている、ポリペプチド。
(g3)請求項10に記載の(b1)から(b23)、および(c3)のいずれかに記載のポリペプチド。 - Emicizumab改変体を製造する方法であって、以下の工程(a)を含む、方法。
(a)以下の(i)~(iii)のうち一以上により置換する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換。 - Emicizumab改変体を単離する方法であって、以下の工程(a)~(c)を含む、方法。
(a)以下の(i)~(iii)のうち一以上によりEmicizumab改変体を産生する工程であって、ここで、番号付けはKabat番号付けシステムに従う、工程;
(i)配列番号:7、8、9に記載の軽鎖CDR1、2、3のアミノ酸配列を含む抗体軽鎖可変ドメインにおいて、K24、A25、S26、R27、N28、I29、E30、R31、Q32、L33、A34、Q50、A51、S52、R53、K54、E55、S56、Q89、Q90、Y91、S92、D93、P94、P95、L96及びT97からなる群から選択される1つ以上のアミノ酸における置換
(ii)配列番号:1、2、3に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、Y31、Y32、D33、I34、Q35、S50、I51、S52、P52a、S53、G54、Q55、S56、T57、Y58、Y59、R60、R61、E62、V63、K64、G65、R95、T96、G97、R98、E99、Y100、G100a、G100b、G100c、W100d、Y100e、F100f、D101およびY102からなる群から選択される1つ以上のアミノ酸における置換
(iii)配列番号:4、5、6に記載の重鎖CDR1、2、3のアミノ酸配列を含む抗体重鎖可変ドメインにおいて、D31、N32、N33、M34、D35、D50、I51、T52a、N52、R53、S54、G55、G56、S57、I58、Y59、N60、E61、E62、F63、Q64、D65、R95、K96、S97、Y98、G99、Y100、Y100a、L100b、D101およびE102からなる群から選択される1つ以上のアミノ酸における置換
(b)(a)において産生された該改変体のFVIII補因子機能代替活性を測定する工程;ならびに
(c)Emicizumabと比べてFVIII補因子機能代替活性が向上したEmicizumab改変体を選択する工程
を包含する、方法。
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CA3027018A CA3027018A1 (en) | 2016-07-29 | 2017-07-27 | Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity |
RU2019104730A RU2821642C2 (ru) | 2016-07-29 | 2017-07-27 | Биспецифическое антитело, обладающее повышенной активностью, альтернативной функции кофактора fviii |
KR1020197005379A KR102591955B1 (ko) | 2016-07-29 | 2017-07-27 | 증강된 fviii 보인자 기능 대체 활성을 갖는 이중특이성 항체 |
US16/318,883 US20190185578A1 (en) | 2016-07-29 | 2017-07-27 | Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity |
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CN201780039875.4A CN109415444B (zh) | 2016-07-29 | 2017-07-27 | 显示增加的备选fviii辅因子功能活性的双特异性抗体 |
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CN202410178980.8A CN117986372A (zh) | 2016-07-29 | 2017-07-27 | 显示增加的备选fviii辅因子功能活性的双特异性抗体 |
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MX2018015721A MX2018015721A (es) | 2016-07-29 | 2017-07-27 | Anticuerpos biespecificos que exhiben actividad de funcion de cofactor fviii alternativa mejorada. |
US17/528,371 US20220073644A1 (en) | 2016-07-29 | 2021-11-17 | Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity |
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JP6496095B1 (ja) * | 2017-09-29 | 2019-04-03 | 中外製薬株式会社 | 血液凝固第viii因子(fviii)補因子機能代替活性を有する多重特異性抗原結合分子および当該分子を有効成分として含有する薬学的製剤 |
WO2019096874A1 (en) | 2017-11-15 | 2019-05-23 | Novo Nordisk A/S | Factor x binders enhancing fx activation |
US10450381B2 (en) | 2010-11-17 | 2019-10-22 | Chugai Seiyaku Kabushiki Kaisha | Methods of treatment that include the administration of bispecific antibodies |
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US11919969B2 (en) | 2017-06-22 | 2024-03-05 | Kymab Limited | Bispecific antibodies for factor IX and factor X |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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Citations (79)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4676980A (en) | 1985-09-23 | 1987-06-30 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Target specific cross-linked heteroantibodies |
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
EP0404097A2 (de) | 1989-06-22 | 1990-12-27 | BEHRINGWERKE Aktiengesellschaft | Bispezifische und oligospezifische, mono- und oligovalente Rezeptoren, ihre Herstellung und Verwendung |
WO1993001161A1 (en) | 1991-07-11 | 1993-01-21 | Pfizer Limited | Process for preparing sertraline intermediates |
WO1993008829A1 (en) | 1991-11-04 | 1993-05-13 | The Regents Of The University Of California | Compositions that mediate killing of hiv-infected cells |
WO1993016185A2 (en) | 1992-02-06 | 1993-08-19 | Creative Biomolecules, Inc. | Biosynthetic binding protein for cancer marker |
WO1994029351A2 (en) | 1993-06-16 | 1994-12-22 | Celltech Limited | Antibodies |
US5500362A (en) | 1987-01-08 | 1996-03-19 | Xoma Corporation | Chimeric antibody with specificity to human B cell surface antigen |
US5571894A (en) | 1991-02-05 | 1996-11-05 | Ciba-Geigy Corporation | Recombinant antibodies specific for a growth factor receptor |
US5587458A (en) | 1991-10-07 | 1996-12-24 | Aronex Pharmaceuticals, Inc. | Anti-erbB-2 antibodies, combinations thereof, and therapeutic and diagnostic uses thereof |
US5624821A (en) | 1987-03-18 | 1997-04-29 | Scotgen Biopharmaceuticals Incorporated | Antibodies with altered effector functions |
US5648237A (en) | 1991-09-19 | 1997-07-15 | Genentech, Inc. | Expression of functional antibody fragments |
WO1997030087A1 (en) | 1996-02-16 | 1997-08-21 | Glaxo Group Limited | Preparation of glycosylated antibodies |
US5731168A (en) | 1995-03-01 | 1998-03-24 | Genentech, Inc. | Method for making heteromultimeric polypeptides |
US5750373A (en) | 1990-12-03 | 1998-05-12 | Genentech, Inc. | Enrichment method for variant proteins having altered binding properties, M13 phagemids, and growth hormone variants |
US5770429A (en) | 1990-08-29 | 1998-06-23 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
US5789199A (en) | 1994-11-03 | 1998-08-04 | Genentech, Inc. | Process for bacterial production of polypeptides |
US5821337A (en) | 1991-06-14 | 1998-10-13 | Genentech, Inc. | Immunoglobulin variants |
US5840523A (en) | 1995-03-01 | 1998-11-24 | Genetech, Inc. | Methods and compositions for secretion of heterologous polypeptides |
WO1998058964A1 (en) | 1997-06-24 | 1998-12-30 | Genentech, Inc. | Methods and compositions for galactosylated glycoproteins |
US5869046A (en) | 1995-04-14 | 1999-02-09 | Genentech, Inc. | Altered polypeptides with increased half-life |
WO1999022764A1 (en) | 1997-10-31 | 1999-05-14 | Genentech, Inc. | Methods and compositions comprising glycoprotein glycoforms |
US5959177A (en) | 1989-10-27 | 1999-09-28 | The Scripps Research Institute | Transgenic plants expressing assembled secretory antibodies |
WO1999051642A1 (en) | 1998-04-02 | 1999-10-14 | Genentech, Inc. | Antibody variants and fragments thereof |
US6040498A (en) | 1998-08-11 | 2000-03-21 | North Caroline State University | Genetically engineered duckweed |
US6075181A (en) | 1990-01-12 | 2000-06-13 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
WO2000061739A1 (en) | 1999-04-09 | 2000-10-19 | Kyowa Hakko Kogyo Co., Ltd. | Method for controlling the activity of immunologically functional molecule |
US6150584A (en) | 1990-01-12 | 2000-11-21 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
US6194551B1 (en) | 1998-04-02 | 2001-02-27 | Genentech, Inc. | Polypeptide variants |
WO2001029246A1 (fr) | 1999-10-19 | 2001-04-26 | Kyowa Hakko Kogyo Co., Ltd. | Procede de production d'un polypeptide |
US6248516B1 (en) | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
WO2002031140A1 (fr) | 2000-10-06 | 2002-04-18 | Kyowa Hakko Kogyo Co., Ltd. | Cellules produisant des compositions d'anticorps |
US6420548B1 (en) | 1999-10-04 | 2002-07-16 | Medicago Inc. | Method for regulating transcription of foreign genes |
US20020164328A1 (en) | 2000-10-06 | 2002-11-07 | Toyohide Shinkawa | Process for purifying antibody |
WO2003011878A2 (en) | 2001-08-03 | 2003-02-13 | Glycart Biotechnology Ag | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
US20030115614A1 (en) | 2000-10-06 | 2003-06-19 | Yutaka Kanda | Antibody composition-producing cell |
US6602684B1 (en) | 1998-04-20 | 2003-08-05 | Glycart Biotechnology Ag | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US20030157108A1 (en) | 2001-10-25 | 2003-08-21 | Genentech, Inc. | Glycoprotein compositions |
WO2003085119A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Procede d'amelioration de l'activite d'une composition d'anticorps de liaison avec le recepteur fc$g(g) iiia |
WO2003085107A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Cellules à génome modifié |
WO2003084570A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Medicament contenant une composition d'anticorps appropriee au patient souffrant de polymorphisme fc$g(g)riiia |
US20040093621A1 (en) | 2001-12-25 | 2004-05-13 | Kyowa Hakko Kogyo Co., Ltd | Antibody composition which specifically binds to CD20 |
US6737056B1 (en) | 1999-01-15 | 2004-05-18 | Genentech, Inc. | Polypeptide variants with altered effector function |
US20040109865A1 (en) | 2002-04-09 | 2004-06-10 | Kyowa Hakko Kogyo Co., Ltd. | Antibody composition-containing medicament |
US20040110282A1 (en) | 2002-04-09 | 2004-06-10 | Kyowa Hakko Kogyo Co., Ltd. | Cells in which activity of the protein involved in transportation of GDP-fucose is reduced or lost |
US20040132140A1 (en) | 2002-04-09 | 2004-07-08 | Kyowa Hakko Kogyo Co., Ltd. | Production process for antibody composition |
WO2004056312A2 (en) | 2002-12-16 | 2004-07-08 | Genentech, Inc. | Immunoglobulin variants and uses thereof |
US20050014934A1 (en) | 2002-10-15 | 2005-01-20 | Hinton Paul R. | Alteration of FcRn binding affinities or serum half-lives of antibodies by mutagenesis |
US20050079574A1 (en) | 2003-01-16 | 2005-04-14 | Genentech, Inc. | Synthetic antibody phage libraries |
WO2005035754A1 (ja) | 2003-10-14 | 2005-04-21 | Chugai Seiyaku Kabushiki Kaisha | 機能蛋白質を代替する二重特異性抗体 |
WO2005035778A1 (ja) | 2003-10-09 | 2005-04-21 | Kyowa Hakko Kogyo Co., Ltd. | α1,6-フコシルトランスフェラーゼの機能を抑制するRNAを用いた抗体組成物の製造法 |
WO2005035586A1 (ja) | 2003-10-08 | 2005-04-21 | Kyowa Hakko Kogyo Co., Ltd. | 融合蛋白質組成物 |
WO2005035756A1 (ja) | 2003-10-10 | 2005-04-21 | Chugai Seiyaku Kabushiki Kaisha | 機能蛋白質を代替する二種特異性抗体 |
US20050119455A1 (en) | 2002-06-03 | 2005-06-02 | Genentech, Inc. | Synthetic antibody phage libraries |
US20050123546A1 (en) | 2003-11-05 | 2005-06-09 | Glycart Biotechnology Ag | Antigen binding molecules with increased Fc receptor binding affinity and effector function |
WO2005053742A1 (ja) | 2003-12-04 | 2005-06-16 | Kyowa Hakko Kogyo Co., Ltd. | 抗体組成物を含有する医薬 |
WO2005100402A1 (en) | 2004-04-13 | 2005-10-27 | F.Hoffmann-La Roche Ag | Anti-p-selectin antibodies |
US20050260186A1 (en) | 2003-03-05 | 2005-11-24 | Halozyme, Inc. | Soluble glycosaminoglycanases and methods of preparing and using soluble glycosaminoglycanases |
US20050266000A1 (en) | 2004-04-09 | 2005-12-01 | Genentech, Inc. | Variable domain library and uses |
WO2006029879A2 (en) | 2004-09-17 | 2006-03-23 | F.Hoffmann-La Roche Ag | Anti-ox40l antibodies |
US7041870B2 (en) | 2000-11-30 | 2006-05-09 | Medarex, Inc. | Transgenic transchromosomal rodents for making human antibodies |
US20060104968A1 (en) | 2003-03-05 | 2006-05-18 | Halozyme, Inc. | Soluble glycosaminoglycanases and methods of preparing and using soluble glycosaminogly ycanases |
WO2006109592A1 (ja) | 2005-04-08 | 2006-10-19 | Chugai Seiyaku Kabushiki Kaisha | 血液凝固第viii因子の機能代替抗体 |
US7125978B1 (en) | 1999-10-04 | 2006-10-24 | Medicago Inc. | Promoter for regulating expression of foreign genes |
US7189826B2 (en) | 1997-11-24 | 2007-03-13 | Institute For Human Genetics And Biochemistry | Monoclonal human natural antibodies |
US20070061900A1 (en) | 2000-10-31 | 2007-03-15 | Murphy Andrew J | Methods of modifying eukaryotic cells |
US20070117126A1 (en) | 1999-12-15 | 2007-05-24 | Genentech, Inc. | Shotgun scanning |
US20070160598A1 (en) | 2005-11-07 | 2007-07-12 | Dennis Mark S | Binding polypeptides with diversified and consensus vh/vl hypervariable sequences |
US20070237764A1 (en) | 2005-12-02 | 2007-10-11 | Genentech, Inc. | Binding polypeptides with restricted diversity sequences |
US20070292936A1 (en) | 2006-05-09 | 2007-12-20 | Genentech, Inc. | Binding polypeptides with optimized scaffolds |
US7371826B2 (en) | 1999-01-15 | 2008-05-13 | Genentech, Inc. | Polypeptide variants with altered effector function |
WO2008077546A1 (en) | 2006-12-22 | 2008-07-03 | F. Hoffmann-La Roche Ag | Antibodies against insulin-like growth factor i receptor and uses thereof |
US20090002360A1 (en) | 2007-05-25 | 2009-01-01 | Innolux Display Corp. | Liquid crystal display device and method for driving same |
WO2009089004A1 (en) | 2008-01-07 | 2009-07-16 | Amgen Inc. | Method for making antibody fc-heterodimeric molecules using electrostatic steering effects |
WO2012067176A1 (ja) | 2010-11-17 | 2012-05-24 | 中外製薬株式会社 | 血液凝固第viii因子の機能を代替する機能を有する多重特異性抗原結合分子 |
WO2013046704A2 (en) | 2011-09-30 | 2013-04-04 | Chugai Seiyaku Kabushiki Kaisha | THERAPEUTIC ANTIGEN-BINDING MOLECULE WITH A FcRn-BINDING DOMAIN THAT PROMOTES ANTIGEN CLEARANCE |
JP2015504434A (ja) * | 2011-11-23 | 2015-02-12 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングBayer Intellectual Property GmbH | 抗fgfr2抗体およびその使用 |
WO2015194233A1 (ja) * | 2014-06-20 | 2015-12-23 | 中外製薬株式会社 | 血液凝固第viii因子および/または活性化血液凝固第viiiの活性の低下ないし欠損によって発症および/または進展する疾患の予防および/または治療に用いられる医薬組成物 |
WO2016001810A1 (en) * | 2014-07-01 | 2016-01-07 | Pfizer Inc. | Bispecific heterodimeric diabodies and uses thereof |
-
2017
- 2017-07-27 MX MX2018015721A patent/MX2018015721A/es unknown
- 2017-07-27 CA CA3027018A patent/CA3027018A1/en active Pending
- 2017-07-27 EP EP17834452.9A patent/EP3492496A4/en active Pending
- 2017-07-27 JP JP2018530374A patent/JP7050677B2/ja active Active
- 2017-07-27 KR KR1020197005379A patent/KR102591955B1/ko active IP Right Grant
- 2017-07-27 CN CN201780039875.4A patent/CN109415444B/zh active Active
- 2017-07-27 WO PCT/JP2017/027152 patent/WO2018021450A1/ja unknown
- 2017-07-27 BR BR112019001179-0A patent/BR112019001179A2/pt unknown
- 2017-07-27 CN CN202410178980.8A patent/CN117986372A/zh active Pending
- 2017-07-27 US US16/318,883 patent/US20190185578A1/en not_active Abandoned
- 2017-07-27 AU AU2017303205A patent/AU2017303205A1/en active Pending
-
2021
- 2021-11-17 US US17/528,371 patent/US20220073644A1/en active Pending
Patent Citations (83)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
US4676980A (en) | 1985-09-23 | 1987-06-30 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Target specific cross-linked heteroantibodies |
US5500362A (en) | 1987-01-08 | 1996-03-19 | Xoma Corporation | Chimeric antibody with specificity to human B cell surface antigen |
US5624821A (en) | 1987-03-18 | 1997-04-29 | Scotgen Biopharmaceuticals Incorporated | Antibodies with altered effector functions |
US5648260A (en) | 1987-03-18 | 1997-07-15 | Scotgen Biopharmaceuticals Incorporated | DNA encoding antibodies with altered effector functions |
US6248516B1 (en) | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
EP0404097A2 (de) | 1989-06-22 | 1990-12-27 | BEHRINGWERKE Aktiengesellschaft | Bispezifische und oligospezifische, mono- und oligovalente Rezeptoren, ihre Herstellung und Verwendung |
US6417429B1 (en) | 1989-10-27 | 2002-07-09 | The Scripps Research Institute | Transgenic plants expressing assembled secretory antibodies |
US5959177A (en) | 1989-10-27 | 1999-09-28 | The Scripps Research Institute | Transgenic plants expressing assembled secretory antibodies |
US6150584A (en) | 1990-01-12 | 2000-11-21 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
US6075181A (en) | 1990-01-12 | 2000-06-13 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
US5770429A (en) | 1990-08-29 | 1998-06-23 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
US5750373A (en) | 1990-12-03 | 1998-05-12 | Genentech, Inc. | Enrichment method for variant proteins having altered binding properties, M13 phagemids, and growth hormone variants |
US5571894A (en) | 1991-02-05 | 1996-11-05 | Ciba-Geigy Corporation | Recombinant antibodies specific for a growth factor receptor |
US5821337A (en) | 1991-06-14 | 1998-10-13 | Genentech, Inc. | Immunoglobulin variants |
WO1993001161A1 (en) | 1991-07-11 | 1993-01-21 | Pfizer Limited | Process for preparing sertraline intermediates |
US5648237A (en) | 1991-09-19 | 1997-07-15 | Genentech, Inc. | Expression of functional antibody fragments |
US5587458A (en) | 1991-10-07 | 1996-12-24 | Aronex Pharmaceuticals, Inc. | Anti-erbB-2 antibodies, combinations thereof, and therapeutic and diagnostic uses thereof |
WO1993008829A1 (en) | 1991-11-04 | 1993-05-13 | The Regents Of The University Of California | Compositions that mediate killing of hiv-infected cells |
WO1993016185A2 (en) | 1992-02-06 | 1993-08-19 | Creative Biomolecules, Inc. | Biosynthetic binding protein for cancer marker |
WO1994029351A2 (en) | 1993-06-16 | 1994-12-22 | Celltech Limited | Antibodies |
US5789199A (en) | 1994-11-03 | 1998-08-04 | Genentech, Inc. | Process for bacterial production of polypeptides |
US5731168A (en) | 1995-03-01 | 1998-03-24 | Genentech, Inc. | Method for making heteromultimeric polypeptides |
US5840523A (en) | 1995-03-01 | 1998-11-24 | Genetech, Inc. | Methods and compositions for secretion of heterologous polypeptides |
US5869046A (en) | 1995-04-14 | 1999-02-09 | Genentech, Inc. | Altered polypeptides with increased half-life |
WO1997030087A1 (en) | 1996-02-16 | 1997-08-21 | Glaxo Group Limited | Preparation of glycosylated antibodies |
WO1998058964A1 (en) | 1997-06-24 | 1998-12-30 | Genentech, Inc. | Methods and compositions for galactosylated glycoproteins |
WO1999022764A1 (en) | 1997-10-31 | 1999-05-14 | Genentech, Inc. | Methods and compositions comprising glycoprotein glycoforms |
US7189826B2 (en) | 1997-11-24 | 2007-03-13 | Institute For Human Genetics And Biochemistry | Monoclonal human natural antibodies |
WO1999051642A1 (en) | 1998-04-02 | 1999-10-14 | Genentech, Inc. | Antibody variants and fragments thereof |
US6194551B1 (en) | 1998-04-02 | 2001-02-27 | Genentech, Inc. | Polypeptide variants |
US6602684B1 (en) | 1998-04-20 | 2003-08-05 | Glycart Biotechnology Ag | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US6040498A (en) | 1998-08-11 | 2000-03-21 | North Caroline State University | Genetically engineered duckweed |
US7332581B2 (en) | 1999-01-15 | 2008-02-19 | Genentech, Inc. | Polypeptide variants with altered effector function |
US7371826B2 (en) | 1999-01-15 | 2008-05-13 | Genentech, Inc. | Polypeptide variants with altered effector function |
US6737056B1 (en) | 1999-01-15 | 2004-05-18 | Genentech, Inc. | Polypeptide variants with altered effector function |
WO2000061739A1 (en) | 1999-04-09 | 2000-10-19 | Kyowa Hakko Kogyo Co., Ltd. | Method for controlling the activity of immunologically functional molecule |
US7125978B1 (en) | 1999-10-04 | 2006-10-24 | Medicago Inc. | Promoter for regulating expression of foreign genes |
US6420548B1 (en) | 1999-10-04 | 2002-07-16 | Medicago Inc. | Method for regulating transcription of foreign genes |
WO2001029246A1 (fr) | 1999-10-19 | 2001-04-26 | Kyowa Hakko Kogyo Co., Ltd. | Procede de production d'un polypeptide |
US20070117126A1 (en) | 1999-12-15 | 2007-05-24 | Genentech, Inc. | Shotgun scanning |
WO2002031140A1 (fr) | 2000-10-06 | 2002-04-18 | Kyowa Hakko Kogyo Co., Ltd. | Cellules produisant des compositions d'anticorps |
US20030115614A1 (en) | 2000-10-06 | 2003-06-19 | Yutaka Kanda | Antibody composition-producing cell |
US20020164328A1 (en) | 2000-10-06 | 2002-11-07 | Toyohide Shinkawa | Process for purifying antibody |
US20070061900A1 (en) | 2000-10-31 | 2007-03-15 | Murphy Andrew J | Methods of modifying eukaryotic cells |
US7041870B2 (en) | 2000-11-30 | 2006-05-09 | Medarex, Inc. | Transgenic transchromosomal rodents for making human antibodies |
WO2003011878A2 (en) | 2001-08-03 | 2003-02-13 | Glycart Biotechnology Ag | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
US20030157108A1 (en) | 2001-10-25 | 2003-08-21 | Genentech, Inc. | Glycoprotein compositions |
US20040093621A1 (en) | 2001-12-25 | 2004-05-13 | Kyowa Hakko Kogyo Co., Ltd | Antibody composition which specifically binds to CD20 |
WO2003085119A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Procede d'amelioration de l'activite d'une composition d'anticorps de liaison avec le recepteur fc$g(g) iiia |
WO2003085107A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Cellules à génome modifié |
WO2003084570A1 (fr) | 2002-04-09 | 2003-10-16 | Kyowa Hakko Kogyo Co., Ltd. | Medicament contenant une composition d'anticorps appropriee au patient souffrant de polymorphisme fc$g(g)riiia |
US20040110704A1 (en) | 2002-04-09 | 2004-06-10 | Kyowa Hakko Kogyo Co., Ltd. | Cells of which genome is modified |
US20040109865A1 (en) | 2002-04-09 | 2004-06-10 | Kyowa Hakko Kogyo Co., Ltd. | Antibody composition-containing medicament |
US20040110282A1 (en) | 2002-04-09 | 2004-06-10 | Kyowa Hakko Kogyo Co., Ltd. | Cells in which activity of the protein involved in transportation of GDP-fucose is reduced or lost |
US20040132140A1 (en) | 2002-04-09 | 2004-07-08 | Kyowa Hakko Kogyo Co., Ltd. | Production process for antibody composition |
US20050119455A1 (en) | 2002-06-03 | 2005-06-02 | Genentech, Inc. | Synthetic antibody phage libraries |
US20050014934A1 (en) | 2002-10-15 | 2005-01-20 | Hinton Paul R. | Alteration of FcRn binding affinities or serum half-lives of antibodies by mutagenesis |
WO2004056312A2 (en) | 2002-12-16 | 2004-07-08 | Genentech, Inc. | Immunoglobulin variants and uses thereof |
US20050079574A1 (en) | 2003-01-16 | 2005-04-14 | Genentech, Inc. | Synthetic antibody phage libraries |
US20050260186A1 (en) | 2003-03-05 | 2005-11-24 | Halozyme, Inc. | Soluble glycosaminoglycanases and methods of preparing and using soluble glycosaminoglycanases |
US20060104968A1 (en) | 2003-03-05 | 2006-05-18 | Halozyme, Inc. | Soluble glycosaminoglycanases and methods of preparing and using soluble glycosaminogly ycanases |
WO2005035586A1 (ja) | 2003-10-08 | 2005-04-21 | Kyowa Hakko Kogyo Co., Ltd. | 融合蛋白質組成物 |
WO2005035778A1 (ja) | 2003-10-09 | 2005-04-21 | Kyowa Hakko Kogyo Co., Ltd. | α1,6-フコシルトランスフェラーゼの機能を抑制するRNAを用いた抗体組成物の製造法 |
WO2005035756A1 (ja) | 2003-10-10 | 2005-04-21 | Chugai Seiyaku Kabushiki Kaisha | 機能蛋白質を代替する二種特異性抗体 |
WO2005035754A1 (ja) | 2003-10-14 | 2005-04-21 | Chugai Seiyaku Kabushiki Kaisha | 機能蛋白質を代替する二重特異性抗体 |
US20050123546A1 (en) | 2003-11-05 | 2005-06-09 | Glycart Biotechnology Ag | Antigen binding molecules with increased Fc receptor binding affinity and effector function |
WO2005053742A1 (ja) | 2003-12-04 | 2005-06-16 | Kyowa Hakko Kogyo Co., Ltd. | 抗体組成物を含有する医薬 |
US20050266000A1 (en) | 2004-04-09 | 2005-12-01 | Genentech, Inc. | Variable domain library and uses |
WO2005100402A1 (en) | 2004-04-13 | 2005-10-27 | F.Hoffmann-La Roche Ag | Anti-p-selectin antibodies |
WO2006029879A2 (en) | 2004-09-17 | 2006-03-23 | F.Hoffmann-La Roche Ag | Anti-ox40l antibodies |
WO2006109592A1 (ja) | 2005-04-08 | 2006-10-19 | Chugai Seiyaku Kabushiki Kaisha | 血液凝固第viii因子の機能代替抗体 |
US20070160598A1 (en) | 2005-11-07 | 2007-07-12 | Dennis Mark S | Binding polypeptides with diversified and consensus vh/vl hypervariable sequences |
US20070237764A1 (en) | 2005-12-02 | 2007-10-11 | Genentech, Inc. | Binding polypeptides with restricted diversity sequences |
US20070292936A1 (en) | 2006-05-09 | 2007-12-20 | Genentech, Inc. | Binding polypeptides with optimized scaffolds |
WO2008077546A1 (en) | 2006-12-22 | 2008-07-03 | F. Hoffmann-La Roche Ag | Antibodies against insulin-like growth factor i receptor and uses thereof |
US20090002360A1 (en) | 2007-05-25 | 2009-01-01 | Innolux Display Corp. | Liquid crystal display device and method for driving same |
WO2009089004A1 (en) | 2008-01-07 | 2009-07-16 | Amgen Inc. | Method for making antibody fc-heterodimeric molecules using electrostatic steering effects |
WO2012067176A1 (ja) | 2010-11-17 | 2012-05-24 | 中外製薬株式会社 | 血液凝固第viii因子の機能を代替する機能を有する多重特異性抗原結合分子 |
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JP2015504434A (ja) * | 2011-11-23 | 2015-02-12 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングBayer Intellectual Property GmbH | 抗fgfr2抗体およびその使用 |
WO2015194233A1 (ja) * | 2014-06-20 | 2015-12-23 | 中外製薬株式会社 | 血液凝固第viii因子および/または活性化血液凝固第viiiの活性の低下ないし欠損によって発症および/または進展する疾患の予防および/または治療に用いられる医薬組成物 |
WO2016001810A1 (en) * | 2014-07-01 | 2016-01-07 | Pfizer Inc. | Bispecific heterodimeric diabodies and uses thereof |
Non-Patent Citations (82)
Title |
---|
"Remington's Pharmaceutical Sciences", 1980 |
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, vol. 275, no. 2, 2000, pages 553 - 557 |
BIOCHIM.BIOPHYS. ACTA, vol. 871, 1986, pages 268 - 278 |
BLOOD, vol. 124, no. 20, 13 November 2014 (2014-11-13), pages 3165 - 71 |
BLOOD, vol. 127, no. 13, 31 March 2016 (2016-03-31), pages 1633 - 1641 |
BLOOD, vol. 58, 1981, pages 1 - 13 |
BOERNER ET AL., J. IMMUNOL., vol. 147, 1991, pages 86 |
BRENNAN ET AL., SCIENCE, vol. 229, 1985, pages 81 |
BRODEUR ET AL.: "Monoclonal Antibody Production Techniques and Applications", 1987, MARCEL DEKKER, INC., pages: 51 - 63 |
BRUGGEMANN, M. ET AL., J. EXP. MED., vol. 166, 1987, pages 1351 - 1361 |
CARDOSO, D.F. ET AL.: "Neutralizing Human Anti Crotoxin scFv Isolated from a Nonimmunized Phage Library, Scand", J. IMMUNOL., vol. 51, no. 4, 2000, pages 337 - 344, XP055580556, ISSN: 0300-9475 * |
CHARLTON: "Methods in Molecular Biology", vol. 248, 2003, HUMANA PRESS, pages: 245 - 254 |
CHOTHIA; LESK, J. MOL. BIOL., vol. 196, 1987, pages 901 - 917 |
CLACKSON ET AL., NATURE, vol. 352, 1991, pages 624 - 628 |
CLARKSON ET AL., NATURE, vol. 352, 1991, pages 624 - 628 |
CLYNES ET AL., PROC. NAT'L ACAD. SCI. USA, vol. 95, 1998, pages 652 - 656 |
CRAGG, M.S. ET AL., BLOOD, vol. 101, 2003, pages 1045 - 1052 |
CRAGG, M.S.; M.J. GLENNIE, BLOOD, vol. 103, 2004, pages 2738 - 2743 |
CUNNINGHAM; WELLS, SCIENCE, vol. 244, 1989, pages 1081 - 1085 |
DUNCAN; WINTER, NATURE, vol. 322, 1988, pages 738 - 40 |
FELLOUSE, PROC. NATL. ACAD. SCI. USA, vol. 101, no. 34, 2004, pages 12467 - 12472 |
FLATMAN ET AL., J. CHROMATOGR. B, vol. 848, 2007, pages 79 - 87 |
GAZZANO-SANTORO ET AL., J. IMMUNOL. METHODS, vol. 202, 1996, pages 163 |
GERNGROSS, NAT. BIOTECH., vol. 22, 2004, pages 1409 - 1414 |
GRAHAM ET AL., J. GEN VIROL., vol. 36, 1977, pages 59 |
GRIFFITHS ET AL., EMBO J, vol. 12, 1993, pages 725 - 734 |
GRIFFITHS, A.D. ET AL.: "Human anti-self antibodies with high specificity from phage display libraries", EMBO J., vol. 12, no. 2, 1993, pages 725 - 734, XP002733039, ISSN: 0261-4189 * |
GRUBER ET AL., J. IMMUNOL., vol. 152, 1994, pages 5368 |
GUYER ET AL., J. IMMUNOL., vol. 117, 1976, pages 587 |
HELLSTROM, I ET AL., PROC. NAT'L ACAD. SCI. USA, vol. 82, 1985, pages 1499 - 1502 |
HELLSTROM, I. ET AL., PROC. NAT'L ACAD. SCI. USA, vol. 83, 1986, pages 7059 - 7063 |
HOLLINGER ET AL., PROC. NAT!. ACAD. SCI. USA, vol. 90, 1993, pages 6444 - 6448 |
HOLLINGER ET AL., PROC. NATL. ACAD. SCI. USA, vol. 90, 1993, pages 6444 - 6448 |
HOOGENBOOM; O'BRIEN ET AL.: "Methods in Molecular Biology", vol. 178, 2001, HUMAN PRESS, pages: 1 - 37 |
HOOGENBOOM; WINTER, J. MOL. BIOL., vol. 227, 1992, pages 381 - 388 |
HUDSON ET AL., NAT. MED., vol. 9, 2003, pages 129 - 134 |
IDUSOGIE ET AL., J. IMMUNOL., vol. 164, 2000, pages 4178 - 4184 |
J THROMB HAEMOST, vol. 12, no. 2, February 2014 (2014-02-01), pages 206 - 213 |
KABAT ET AL.: "Sequences of Proteins of Immunological Interest", 1991, PUBLIC HEALTH SERVICE |
KANDA, Y. ET AL., BIOTECHNOL. BIOENG., vol. 94, no. 4, 2006, pages 680 - 688 |
KIM ET AL., J. IMMUNOL., vol. 24, 1994, pages 249 |
KOSTELNY ET AL., J. IMMUNOL., vol. 148, no. 5, 1992, pages 1547 - 1553 |
KOZBOR, IMMUNOL., vol. 133, 1984, pages 3001 |
LEE ET AL., 7. MOL. BIOL., vol. 340, no. 5, 2004, pages 1073 - 1093 |
LEE ET AL., J. IMMUNOL. METHODS, vol. 284, no. 1-2, 2004, pages 119 - 132 |
LI ET AL., NAT. BIOTECH., vol. 24, 2006, pages 210 - 215 |
LI ET AL., PROC. NATL. ACAD. SCI. USA, vol. 103, 2006, pages 3557 - 3562 |
LONBERG, CURR. OPIN. IMMUNOL., vol. 20, 2008, pages 450 - 459 |
LONBERG, NAT. BIOTECH., vol. 23, 2005, pages 1117 - 1125 |
MABS, vol. 7, no. l, 2015, pages 120 - 8 |
MACCALLUM ET AL., J. MOL. BIOL., vol. 262, 1996, pages 732 - 745 |
MARKS ET AL., J. MOL. BIOL., vol. 222, 1992, pages 581 - 597 |
MARKS; BRADBURY: "Methods in Molecular Biology", vol. 248, 2003, HUMAN PRESS, pages: 161 - 175 |
MATHER ET AL., ANNALS N.Y. ACAD. SCI., vol. 383, 1982, pages 44 - 68 |
MATHER, BIOL. REPROD., vol. 23, 1980, pages 243 - 251 |
MCCAFFERTY ET AL., NATURE, vol. 348, pages 552 - 554 |
MILSTEIN; CUELLO, NATURE, vol. 305, 1983, pages 537 |
NAT MED., vol. 18, no. 10, October 2012 (2012-10-01), pages 1570 - 4 |
NATURE, vol. 312, 1984, pages 330 - 337 |
NATURE, vol. 312, 1984, pages 337 - 342 |
NEW ENG J MED, vol. 374, no. 21, 26 May 2016 (2016-05-26), pages 2044 - 2053 |
NI, XIANDAI MIANYIXUE, vol. 26, no. 4, 2006, pages 265 - 268 |
OKAZAKI ET AL., J. MOL. BIOI., vol. 336, 2004, pages 1239 - 1249 |
PETKOVA, S.B. ET AL., INT L. IMMUNOL., vol. 18, no. 12, 2006, pages 1759 - 1769 |
PLOS ONE, vol. 8, no. 2, 2013, pages e57479 |
PLUCKTHIIN: "The Pharmacology of Monoclonal Antibodies", vol. 113, 1994, SPRINGER-VERLAG, pages: 269 - 315 |
PORTOLANO ET AL., J. IMMUNOL., vol. 150, 1993, pages 880 - 887 |
RAVETCH; KINET, ANNU. REV. IMMUNOL., vol. 9, 1991, pages 457 - 492 |
RIPKA ET AL., ARCH. BIOCHEM. BIOPHYS., vol. 249, 1986, pages 533 - 545 |
ROSEN S; ANDERSSON M; BLOMBA'CK M ET AL.: "Clinical applications of a chromogenic substrate method for determination of FVIII activity", THROMB HAEMOST, vol. 54, 1985, pages 811 - 23 |
SHIELDS ET AL., J. BIOL. CHEM., vol. 9, no. 2, 2001, pages 6591 - 6604 |
SIDHU ET AL., J. MOL. BIOL., vol. 338, no. 2, 2004, pages 299 - 310 |
TRAUNECKER ET AL., EMBO J., vol. 10, 1991, pages 3655 |
TUTT ET AL., J. IMMUNOL., vol. 147, 1991, pages 60 |
URLAUB ET AL., PROC. NATL. ACAD. SCI. USA, vol. 77, 1980, pages 4216 |
VAN DIJK; VAN DE WINKEL, CURT: OPIN. PHARMACOL., vol. 5, 2001, pages 368 - 74 |
VOLLMERS; BRANDLEIN, HISTOLOGY AND HISTOPATHOLOGY, vol. 20, no. 3, 2005, pages 927 - 937 |
VOLLMERS; BRANDLEIN, METHODS AND FINDINGS IN EXPERIMENTAL AND CLINICAL PHARMACOLOGY, vol. 27, no. 3, 2005, pages 185 - 91 |
WINTER ET AL., ANN. REV. IMMUNOL, vol. 12, 1994, pages 433 - 455 |
WRIGHT ET AL., TIBTECH, vol. 15, 1997, pages 26 - 32 |
YAMANE-OHNUKI ET AL., BIOTECH. BIOENG., vol. 87, 2004, pages 614 |
YAZAKI; WU: "Methods in Molecular Biology", vol. 248, 2003, HUMANA PRESS, pages: 255 - 268 |
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