Therapeutic anti-CD40 ligand antibody

Abstract

The present invention provides pharmaceutical compositions for use in methods for treating subjects with anti-CD40 antibodies and CD40L-related diseases or disorders. [Solution] The present invention provides an anti-CD40L antibody and its antigen-binding fragment for treating CD40L-related diseases or disorders, a composition comprising such antibody or antigen-binding fragment, an anti-CD40L antibody with low effector function, and a method of using the same.

Description

Technical Field

[0001] Cross - reference to Related Applications This PCT application claims the benefit of U.S. Provisional Patent Application No. 62 / 510,471, filed on May 24, 2017, the disclosure of which is hereby incorporated by reference in its entirety.

[0002] Sequence Listing This application incorporates by reference in its entirety a Sequence Listing entitled "224823 - 431206_ST25.txt", created on May 23, 2018 at 2:40 PM and 100 KB, which was electronically submitted together at the time of filing.

[0003] Technical Field Anti - CD40L antibodies for treating CD40L - related diseases or disorders, compositions comprising such antibodies, and methods of using them.

Background Art

[0004] The interaction between CD40 and its ligand, CD40L, plays a crucial role in regulating the immune response. CD40L binding to CD40 activates the CD40 pathway, upregulating costimulatory molecules such as CD80 and CD86. Blocking the interaction between CD40 and CD40L with monoclonal antibodies has been shown in various preclinical models to protect against autoimmunity and graft rejection. Recently, in a mouse model of amyotrophic lateral sclerosis (ALS), CD40L-targeting antibodies were shown to delay disease onset and extend post-disease survival (U.S. Patent No. 8,435,514, incorporated herein by reference). Early clinical trials showed efficacy of the humanized anti-CD40L antibody hu5c8 in patients with lupus and immune thrombocytopenic purpura. However, further trials were discontinued due to thromboembolism in patients treated with hu5c8. Furthermore, in vitro and animal preclinical studies have confirmed that the interaction between Fc and the Fc receptor FcγRIIa triggers platelet activation and subsequent aggregation, leading to thromboembolic events. Additionally, it has been reported that Fc binding to complement can suppress or interfere with the induction of immune tolerance. Various attempts have been made to suppress or eliminate the interaction between the Fc region of immunoglobulins and FcγRIIa, and / or the interaction with complement. For example, anti-CD40L antibodies lacking Fc effector function have been created by introducing point mutations (multiple possible) into the Fc region. Other attempts involve using antibody fragments lacking the Fc region, or antibodies containing multiple amino acid substitutions in the Fc region. While the anti-CD40L antibody hu5c8 demonstrated efficacy in human patients, no anti-CD40L antibodies are currently available on the market. Therefore, there is a need for an improved anti-CD40L antibody for human administration that is effective, does not cause platelet activation or aggregation, does not bind to complement, yet is still stable, and binds to CD40L. [Overview of the project]

[0005] The novel antibody polypeptides and their antigen-binding fragments of the present invention provide such improved anti-CD40L antibodies. The following sections merely summarize and are not intended to limit the particular aspects of this disclosure. These aspects and other aspects and embodiments are described in further detail below. All references cited herein are incorporated herein by reference in their entirety. In the event of any conflict between the express disclosure herein and the references incorporated by reference, the express disclosure herein shall prevail.

[0006] The present invention provides isolated, engineered, non-spontaneously occurring antibodies and their antigen-binding fragments that bind to human CD40L and block the binding of CD40 to CD40L. The antibodies and their antigen-binding fragments are engineered to have desired activity and binding. In some embodiments, the antibodies or their antigen-binding fragments are engineered to have attenuated Fc effector function compared to some of the aforementioned anti-CD40L antibodies. The antibodies and their antigen-binding fragments disclosed herein are useful for treating diseases involving CD40L activation, including neurodegenerative or neuromuscular diseases or disorders, inflammatory or immune diseases or disorders, and autoimmune diseases.

[0007] In one embodiment, the present disclosure provides an isolated antibody or an antigen-binding fragment thereof that specifically binds to CD40L, comprising a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, 11, 12, 13, or 14, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16 or 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18 or 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20.

[0008] In some embodiments of the above aspects of the present disclosure, an isolated antibody or an antigen-binding fragment thereof that specifically binds to CD40L is provided, which comprises (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, 11, 12, 13, or 14, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16 or 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18 or 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0009] The disclosure also provides an isolated antibody or its antigen-binding fragment that specifically binds to CD40L, which comprises (a) a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1, 2, 3, or 4, and (b) a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5, 6, 7, or 8.

[0010] In some embodiments, the isolated antibody or its antigen-binding fragment according to the present disclosure includes an Fc region, which is manipulated to suppress or eliminate one or more Fc effector functions. In some embodiments, the isolated antibody is of the IgG1 isotype, and the heavy chain constant region includes the amino acid sequence described in SEQ ID NO: 21. In other embodiments, the antibody includes a heavy chain constant region including the amino acid sequence described in SEQ ID NO: 22. In yet another embodiment, the antibody includes a heavy chain constant region including the amino acid sequence described in SEQ ID NO: 23. In yet another embodiment, the antibody includes a heavy chain constant region including the amino acid sequence described in SEQ ID NO: 24.

[0011] Another aspect of the Disclosure provides a method for treating a subject having a disease or disorder related to CD40L, comprising administering to such subject a therapeutically effective amount of antibody or antibody fragment in accordance with the Disclosure. Another aspect of the Disclosure provides a method for inhibiting an immune response in a subject, comprising administering to such subject a therapeutically effective amount of antibody or antibody fragment in accordance with the Disclosure. [Brief explanation of the drawing]

[0012] [Figure 1-1] Figure 1A is a bar graph showing the titer ranking of 16 antibody clones against the anti-CD40L antibody 5c8. The titer ranking is calculated by dividing the clone's IC50 by the IC50 of 5c8 and multiplying by 100. Figure 1B is a bar graph showing the titer ranking of 16 antibody clones against the anti-CD40L antibody AT-1501. The titer ranking is calculated by dividing the clone's IC50 by the IC50 of AT-1501 and multiplying by 100. [Figure 1-2] Same as above. [Figure 2-1]Figure 2A shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2B shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2C shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2D shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2E shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2F shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2G shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2I shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2J shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2K shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2L shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2M shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2N shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2O shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2P shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2Q shows the antibody binding curves for each clone and the binding curve for 5c8. Figure 2H shows the binding curves for AT-1501 and 5c8. [Figure 2-2] Same as above. [Figure 2-3] Same as above. [Figure 2-4] Same as above. [Figure 2-5] Same as above. [Figure 2-6] Same as above. [Figure 2-7] Same as above. [Figure 2-8] Same as above. [Figure 2-9] Same as above. [Figure 3-1]Figure 3A shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3B shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3C shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3D shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3E shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3F shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3G shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3I shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3J shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3K shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3L shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3M shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3N shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3O shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3P shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3Q shows the antibody binding curves for each clone and the binding curve for AT-1501. Figure 3H shows the binding curves for AT-1501 and 5c8. [Figure 3-2] Same as above. [Figure 3-3] Same as above. [Figure 3-4] Same as above. [Figure 3-5] Same as above. [Figure 3-6] Same as above. [Figure 3-7] Same as above. [Figure 3-8] Same as above. [Figure 3-9] Same as above. [Figure 4]Figure 4A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRI, respectively. The only antibody with significant binding is the 5c8 antibody. Figure 4B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRI, respectively. The only antibody with significant binding is the 5c8 antibody. [Figure 5] Figure 5A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIa, respectively. The only antibody with significant binding is the 5c8 antibody. Figure 5B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIa, respectively. The only antibody with significant binding is the 5c8 antibody. [Figure 6-1] Figure 6A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIa, respectively. None of the antibodies showed significant binding to FcγRIIIa. Figure 6B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIa, respectively. None of the antibodies showed significant binding to FcγRIIIa. Figure 6C is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIb, respectively. None of the antibodies showed significant binding to FcγRIIIb. Figure 6D is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to FcγRIIIb, respectively. None of the antibodies showed significant binding to FcγRIIIb. [Figure 6-2] Same as above. [Figure 7]Figure 7A is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to C1q, respectively. The only antibody with significant binding to C1q was the 5c8 antibody. Figure 7B is a graph showing the binding of eight of the antibody clones, 5c8, AT-1501, and abatacept, to C1q, respectively. The only antibody with significant binding to C1q was the 5c8 antibody.

BEST MODE FOR CARRYING OUT THE INVENTION

[0013] Definitions

[0014] For convenience, the meanings of certain terms and phrases used in the specification, examples, and appended claims are set forth below.

[0015] Terms such as "comprises", "comprised", "comprising", "contains", "containing", etc. have the meaning under the United States Patent Law. These terms are inclusive or non-limiting and do not exclude additional recited elements or method steps. Terms such as "consisting essentially of" and "consists essentially of" have the meaning arising from these terms in the United States Patent Law, and these terms allow for the inclusion of additional ingredients or steps that do not substantially affect the basic and novel features of the claimed invention. The terms "consists of" and "consisting of" have the meaning arising from these terms in the United States Patent Law, and these terms are limiting.

[0016] TMB is an abbreviation for 3,3′,5,5′-tetramethylbenzidine.

[0017] As used herein, the "CDR domain" means an antibody complementarity-determining region, regardless of the presence of adjacent sequences.

[0018] The present invention, as described herein by example, may be suitably carried out without any elements(s) or limitations(s) not specifically disclosed herein.

[0019] "Effector function" refers to the biological activity resulting from the Fc region of an antibody, and it varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cell-mediated cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation. One effector function is the ability of the antibody's Fc region or constant region to bind to a protein, such as Fc receptors (FcRs) (e.g., high-affinity IgG Fc region receptor Ia (FCγRIa) (CD64) (SEQ ID NO: 34), low-affinity immunoglobulin gamma Fc region acceptor IIa (FCγRIIa) (CD32) (SEQ ID NO: 35), low-affinity immunoglobulin gamma Fc region receptor IIIa (FCγRIIIA) (CD16a) (SEQ ID NO: 36), low-affinity immunoglobulin gamma Fc region receptor IIIb (FCγRIIIb) (CD16b) (SEQ ID NO: 37)), but not limited to these. In embodiments of the present invention in which the antibody and its antigen-binding fragment have an Fc domain, such Fc domain is manipulated to suppress or eliminate one or more Fc effector functions. In preferred embodiments, the Fc domain is manipulated to suppress or eliminate the risk of platelet activation and / or platelet aggregation and concomitant thromboembolism.

[0020] As used herein, the singular forms "a," "an," and "the" refer to multiple objects unless otherwise specifically indicated by the context. The use of "or" or "and" means "and / or" unless otherwise specified. Furthermore, the use of the term "including" and other forms such as "include," "includes," and "included" is not limited to these.

[0021] The conjunction "or" is used with the same meaning as "at least one." For example, if a composition contains A or B, the method must contain at least one of A and B, but may contain both A and B. Similarly, if a composition contains "A, B, C or D," it must contain at least one of the group A, B, C, and D, but may contain all of A, B, C, and D or any combination thereof.

[0022] The term "approximately" indicates that a certain degree of minimal imperfection is acceptable in the stated numerical value (approximating to some extent to its exact value; roughly or very close to it; almost the exact value). In contrast to this usual meaning, unless the imperfection given by "approximately" is understood to have a different meaning in the art, as used herein, "approximately" encompasses a variation of up to ±10% from the specified value. Unless otherwise specified, any numbers used herein to represent quantities of components, properties such as molecular weight, reaction conditions, etc., should be understood as being modifiable by the term "approximately."

[0023] CD40L is also known as CD154, gp39, T-BAM, 5c8 antigen, or TNF-related activating protein (TRAP).

[0024] The terms "to treat" and "treatment" include both therapeutic and preventive measures. Therapeutic measures refer to treatment of subjects who have signs or symptoms of a disease, condition, or disorder that should be treated. Preventive measures refer to treatment of subjects who are susceptible to such a disease, condition, or disorder but do not show overt signs of the disease, condition, or disorder. Therefore, treatment may result in the cessation, partial or complete reduction, or suppression of signs or symptoms of the disease, including, but not limited to, extension of survival and cure.

[0025] An "effective dose" of a drug, such as a pharmaceutical preparation, refers to the amount necessary for the required duration and quantity that is effective in achieving the desired therapeutic or preventive outcome.

[0026] "Affinity" refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, as used herein, "binding affinity" refers to intrinsic binding affinity, reflecting the 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y may generally be expressed by the equilibrium dissociation constant (KD), which is the koff / kon ratio between the antibody and its antigen. KD and affinity are inversely correlated. Since the KD value is related to the antibody concentration (the amount of antibody required for a particular experiment), a lower KD value (lower concentration) indicates higher antibody affinity. Affinity may be measured by general methods known in the art, including the methods described herein. In certain specific and exemplary embodiments for measuring binding affinity, measurement may be performed by radioimmunoassay (RIA), or by surface plasmon resonance (SPR) on a BIAcore® instrument (GE Healthcare Europe GmbH, Glattbrugg, Switzerland) by capturing the antibody on a protein A-binding CM5 research-grade sensor chip (GE Healthcare Europe GmbH, Glattbrugg, Switzerland; BR-1000-14) using a human CD40 ligand polypeptide as an analyte. Other methods may include radioimmunoassay and binding equilibrium exclusion. Binding equilibrium exclusion is a general-purpose immunoassay platform capable of measuring equilibrium-dissociation constants, as well as the association and dissociation rate constants of antigen / antibody interactions.

[0027] This specification refers to the percentage of identity between polypeptide or amino acid sequences. The percentage of identity between two sequences is a function of the number of identical positions shared by the two sequences, taking into account the number of gaps and the length of each gap, which must be introduced for optimal alignment of the two sequences. Identity can be measured as either “local identity” or “global identity.” Local identity refers to the degree of sequence relevance between polypeptides, determined by the matching of strings of such sequences. Global identity refers to the degree of sequence relevance of a polypeptide compared to the full length of a reference polypeptide. Unless otherwise specified, identity as used herein means global identity. For the purposes of this disclosure and claims, the percentage of global identity is calculated using the Needleman and Wunsch ((1970) J.Mol.Biol.48:444~453) algorithm, which uses a Blossum62 score matrix with a gap penalty of 12, a gap elongation penalty of 4, and a frameshift gap penalty of 5. For example, there are numerous publicly available software programs that incorporate the Needleman and Wunsch algorithm, such as the GAP program in the GCG software package.

[0028] Those skilled in the art will understand that starting materials, biological and chemical materials, biological and chemical reagents, synthesis methods, purification methods, analytical methods, assay methods, and biological methods other than those specifically exemplified can be incorporated into the practice of the present invention without excessive experimentation. Any functional equivalents of such materials and methods known in the art are intended to be included in this disclosure.

[0029] By convention, amino acid substitutions are indicated by identifying the original amino acid, its position in the specified sequence, and the substituted amino acid. For example, C11S indicates that the cysteine ​​at position 11 of the polypeptide sequence is replaced with serine.

[0030] Humanized antibodies are antibodies produced by non-human species whose protein sequences have been modified to resemble naturally occurring antibody variants in humans. The "humanization" process is typically applied to monoclonal antibodies intended for administration to humans (for example, antibodies created as anticancer drugs).

[0031] Currently, it is common practice to humanize non-human antibodies by inserting relevant CDRs obtained from antibodies created in non-human animals into a human antibody "scaffold." However, the "direct" creation of humanized antibodies can be achieved by inserting an appropriate CDR coding segment (responsible for the desired binding properties) into a human antibody "scaffold." This can be achieved by recombinant DNA expression in mammalian cells using a suitable vector. That is, after creating an antibody in a mouse (or other non-human) with the desired properties, the DNA encoding the antibody can be isolated, cloned into a vector, and sequenced. Subsequently, the DNA sequence corresponding to the antibody's CDR can be determined. Once the precise sequence of the desired CDR is known, a strategy can be devised for appropriately inserting these sequences into a construct containing DNA for human antibody variants. Prior to insertion into the scaffold, the CDR may be modified, for example, to increase specificity.

[0032] The term "human" antibody refers to an antibody that has an amino acid sequence corresponding to an antibody produced in humans, and / or an antibody produced using any method for producing human antibodies. This definition of human antibody explicitly excludes humanized antibodies that contain non-human antigen-binding residues.

[0033] An "antigen-binding antibody fragment" refers to a molecule other than an intact antibody, and includes a portion of an intact antibody that binds to that antigen, and such portion binds to that antigen. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fd fragments, dAb fragment, Fab'-SH, F(ab')2; diabodies; triabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments and minimal recognition units consisting of amino acid residues that mimic the hypervariable region of an antibody (e.g., isolated complementarity-determining regions (CDRs) such as the CDR3 peptide). The antigen-binding fragments disclosed in this application bind to the antigen CD40L.

[0034] Antibody antigen-binding fragments typically contain at least one variable domain. The variable domain may be of any size and have any amino acid composition, and generally contains at least one CDR adjacent to or within a frame having one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains may be positioned relative to each other in an appropriate configuration. For example, the variable region may be a dimer, containing dimers such as VH-VH, VH-VL, or VL-VL. Alternatively, antibody antigen-binding fragments may contain monomeric VH or VL domains.

[0035] The isolated antibodies include bispecific antibodies, where each arm or antigen-binding fragment of the antibody binds to a different target or epitope.

[0036] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably in this specification and refer to antibodies that have a structure substantially similar to that of a natural antibody or that have a heavy chain containing an Fc region.

[0037] "5c8" refers to a mouse anti-human antibody, which conjugates to CD40L and is produced by a hybridoma described in U.S. Patent No. 5,474,771, available from ATCC with accession number HB10916. "hu5c8" refers to a humanized form of 5c8, the sequence of which is disclosed in Karpusas, et al., Structure vol.9, pp321-329, (2001).

[0038] The terms "subject," "individual," and "patient" are used interchangeably in this specification and refer to a human subject, individual, or patient.

[0039] All compositional proportions herein are weight-based proportions of the total composition unless otherwise specified. As used herein, the term “includes” and its variations are intended to be non-limiting, and references to items in the list do not preclude other similar items, which may be useful in the materials, compositions, devices, and methods of the present art. Similarly, the terms “may” and “may comprise” and their variations are intended to be non-limiting, and where an embodiment may or may comprise certain elements or features, other embodiments of the present art that do not contain those elements or features are not excluded.

[0040] The term “isolated protein” or “isolated polypeptide” (e.g., isolated antibody or isolated antigen-binding fragment) refers to a protein or polypeptide that, based on its origin or derivative source, is not associated with the naturally occurring associated components found in its native state; substantially free from other proteins of the same species; expressed by cells obtained from a different species; or does not occur in nature. Therefore, a polypeptide synthesized chemically, or synthesized in a cell system different from that of cells of natural origin, is “isolated” from its naturally occurring associated components. A protein may be made substantially free from its naturally occurring associated components by isolation using protein purification techniques well known in the art.

[0041] As used herein, the term “mutant” is defined as a modified or altered form of the wild-type sequence, in which case, for example, one or more amino acids may be substituted with other amino acids or non-amino acids that do not substantially affect the function. In some embodiments, the mutant may contain an altered side chain for at least one amino acid residue.

[0042] As used herein, the term “antigen” is defined as an entity that can stimulate the production of antibodies, specifically bind to those antibodies, and / or trigger an immune system response. For example, a cell surface protein or a specific linear or nonlinear portion thereof. The term may be abbreviated as “Ag” as used herein.

[0043] Terms such as "specifically binding" mean that an antibody or its antigen-binding fragment forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding may be characterized by an equilibrium dissociation constant (KD) of approximately 3000 nM or less (i.e., a smaller KD means a tighter binding), approximately 2000 nM or less, approximately 1000 nM or less, approximately 500 nM or less, approximately 300 nM or less, approximately 200 nM or less, approximately 100 nM or less, approximately 50 nM or less, approximately 1 nM or less, or approximately 0.5 nM.

[0044] Specific binding to a particular antigen or epitope can be demonstrated, for example, by an antibody whose KD for the antigen or epitope is at least about 1 × 10⁻⁴ M, at least about 1 × 10⁻⁵ M, at least about 1 × 10⁻⁶ M, at least about 1 × 10⁻⁷ M, at least about 1 × 10⁻⁸ M, at least about 1 × 10⁻⁹ M, and in other cases at least about 1 × 10⁻¹⁰ M, at least about 1 × 10⁻¹¹ M, at least about 1 × 10⁻¹² M, or more, in which case KD refers to the equilibrium dissociation constant of the particular antibody-antigen interaction. Typically, an antibody that specifically binds to a certain antigen will have a KD 20 times, 50 times, 100 times, 500 times, 1000 times, 10000 times, 5,000 times, 10,000 times, or more greater against a control molecule compared to against that antigen or epitope. Furthermore, specific binding to a particular antigen or epitope may be demonstrated, for example, by an antibody whose Ka for that antigen or epitope is at least 20, 50, 100, 500, 1000, 5,000, 10,000 or more times greater than that for the control epitope, in which case Ka refers to the association rate of the specific antibody-antigen interaction.

[0045] The term "neutralizing antibody" includes antibodies that can inhibit and / or neutralize the biological activity of CD40L, such as anti-CD40L antibodies or their antigenic fragments that inhibit, prevent or reduce the binding of CD40L to CD40, thereby inhibiting or reducing the signaling pathway evoked by CD40L, and / or inhibiting or reducing the binding of CD40L to CD40.

[0046] The terms “antagonistic antibody” or “antagonistic antibody” are used interchangeably herein and include antibodies that can inhibit and / or neutralize the biological signaling activity of CD40L, as described in the neutralizing antibody section above.

[0047] The terms and expressions used herein are intended to be descriptive rather than restrictive, and in using such terms and expressions, no equivalent or part thereof of any features shown or described is intended to be excluded, but it is recognized that various modifications are possible within the scope of the claimed invention. Accordingly, while aspects of the invention are specifically disclosed by various embodiments which may include preferred embodiments, exemplary embodiments, and optional features, it should be understood that modifications and variations of the concepts disclosed herein may be made by those skilled in the art. Such modifications and variations are deemed to be within the scope of embodiments of the invention which may be described and defined in the appended claims.

[0048] Pharmaceutical composition and administration method

[0049] Pharmaceutical compositions used in accordance with the methods of this disclosure may be formulated in a conventional manner using one or more physiologically acceptable carriers. The pharmacologically acceptable carriers are determined in part by the specific composition being administered and by the specific method used to administer the composition. Therefore, the formulations of compounds useful in the methods of this disclosure are diverse (see, for example, Remington: The Science and Practice of Pharmacy, 20th ed., Gennaro et al. Eds., Lippincott Williams and Wilkins, 2000).

[0050] Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes to make the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspensions, solubilizers, thickeners, stabilizers, and preservatives. Exemplary injectable or infusion additives may include mannitol, citrate monohydrate, sodium hydrogen phosphate dihydrate, sodium dihydrogen phosphate dihydrate, polysorbate 80, sodium chloride, sodium citrate, and water.

[0051] In accordance with this disclosure, the compounds can be administered by any suitable means, which may vary depending on the type of disorder being treated and the properties of the compound itself. For example, in the case of the antibodies and their antigen-binding fragments of the present invention, the administration routes preferably include parenteral administration routes, such as intramuscular, intravenous, intra-arterial, intraperitoneal, intracerebrospinal, intraspinal, epidural, and subcutaneous routes, or administration by sustained-release systems or implantable tablets. Preferably, parenteral administration is performed by injection, most preferably by intravenous, intramuscular, or subcutaneous injection.

[0052] In some embodiments, the anti-CD40L antibody or its antigen-binding fragment is formulated, but before use, it may be lyophilized for storage and reconstituted in appropriate additives according to lyophilization and reconstitution methods known in the art.

[0053] In one exemplary pharmaceutical composition containing an anti-CD40L antibody or its antigen-binding fragment, the composition is formulated for intravenous or subcutaneous administration as a preservative-free sterile solution of the anti-CD40L antibody or its antigen-binding fragment. The formulation may be supplied as a single-use pre-filled pen, as a single-use, for example, a pre-filled glass syringe containing about 1 mL, or as a single-use vial for medical use. Preferably, the pharmaceutical composition containing the anti-CD40L antibody or its antigen-binding fragment is clear, colorless, and has a pH in the range of about 5.0 to about 6.9, preferably about 5.0 to about 6.5, and more preferably about 5.0 to about 6.0. In various embodiments, a formulation containing a pharmaceutical composition may contain, when reconstituted and administered to a subject, about 500 mg to about 1 mg, or about 400 mg to about 10 mg, or about 300 mg to about 30 mg, or about 200 mg to about 50 mg of anti-CD40L antibody or its antigen-binding fragment per 1 mL of solution.

[0054] The selection of an administration regimen for a particular therapeutic agent depends on several factors, including the metabolic rate of the agent in serum or tissue, the level of symptoms, the immunogenicity of the agent, and the reachability of target cells within the biological matrix, the patient's overall health status, and the patient's medical history. Preferably, in a given administration regimen, the amount of therapeutic agent delivered to the patient should be maximized according to the acceptable level of side effects. Therefore, the amount delivered biologically depends, in part, on the individual agent and the severity of the condition being treated. It should be understood that the determination of the appropriate dosage form, dose, and route of administration is within the scope of those skilled in the art in the pharmaceutical and medical fields.

[0055] dose

[0056] The pharmaceutical formulations of this disclosure may contain about 0.001 to about 200 mg / kg of anti-CD40L antibody or its antigen-binding fragment. For example, the anti-CD40L antibody or its antigen-binding fragment may be administered as a bolus by intravenous injection at a dose of about 0.001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 50 mg / kg, or about 0.001 mg / kg to about 10 mg / kg, with the remaining antibody dose administered by intravenous injection. The prescribed dose of anti-CD40L antibody or its antigen-binding fragment may be administered over a period of time, for example, 1 to 2 hours to 5 hours.

[0057] In further embodiments, a portion of the antibody preparation may be administered by subcutaneous injection and / or bolus injection, with the remainder administered by infusion. In some exemplary doses, the antibody preparation may be administered subcutaneously in doses ranging from about 0.001 to about 200 mg / kg, for example, by intravenous injection of anti-CD40L antibody or its antigen-binding fragment in doses ranging from about 0.001 mg / kg to about 100 mg / kg, or from about 0.001 mg / kg to about 50 mg / kg, or from about 0.001 mg / kg to about 10 mg / kg. In some embodiments, the dose may be given as a bolus, with the remaining antibody dose administered by subcutaneous or intravenous injection. A predetermined dose of anti-CD40L antibody or its antigen-binding fragment may be administered over a period of time, for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, or longer.

[0058] Combination therapy

[0059] The antibodies or antibody fragments described herein can be administered alone (as monotherapy) or in combination, i.e., in combination with other agents. For example, in one embodiment, the combination therapy may include one or more additional therapeutic agents. In another embodiment, the combination therapy includes a standard therapeutic treatment (essentially consisting of an antibody or antibody fragment) which may or may not include additional therapeutic agents.

[0060] Adjunctive or concomitant administration (simultaneous administration) includes the simultaneous administration of one of the antibodies or antigen-binding fragments described herein with one or more drugs in the same or different dosage forms, or the individual administration (e.g., sequential administration) of the polypeptide and one or more drugs. Preferably, such concomitant or sequential administration ensures that both the polypeptide and one or more drugs are present in the treated patient's body at the same time.

[0061] Kits and manufactured products

[0062] Furthermore, this specification provides kits containing antibodies or antigen-binding fragments thereof and instructions for use. A kit typically includes a package containing a predetermined combination of reagents along with instructions for use, and a label indicating the intended use of the kit contents. The term "label" or "instructions for use" includes any written or recorded material on or supplied with the kit; otherwise, a label or instructions for use is attached to the kit at any time during the manufacture, transport, sale, or use of the kit. This may take the form prescribed by a government agency regulating the manufacture, use, or sale of a pharmaceutical or biological product, and such notification reflects approval by such agency for manufacture, use, or sale for human administration or animal use. "Label" or "instructions for use" may also include advertising brochures and pamphlets, packaging materials, and audio or video instructions for use.

[0063] Embodiment

[0064] All embodiments of the isolated antibody or its antigen-binding fragment bind to CD40L and inhibit or block the binding of CD40L to CD40. As used herein, “blocking the binding of CD40L to CD40” and “blocking the interaction of CD40L to CD40” are used interchangeably. The inhibition or blockage of binding may be direct or indirect. Generally, the antibody or its antigen-binding fragment physically prevents the binding of CD40L to CD40 by directly and specifically competing with CD40 for the same binding site on CD40L, or by steric hindrance caused by the antibody or its antigen-binding fragment binding in close proximity to the CD40 binding site on CD40L. In other cases, the effect is indirect, for example, by causing an allosteric change in the conformation of CD40L, thereby inhibiting or eliminating the binding of CD40L to CD40.

[0065] One embodiment is an isolated antibody bound to CD40L and comprising a light chain and a heavy chain, wherein the light chain comprises a light chain variable region containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, and the heavy chain comprises a heavy chain variable region, wherein the heavy chain variable region contains an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

[0066] In one embodiment, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising a heavy chain variable region (VH) comprising a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, 11, 12, 13, or 14, and a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and a light chain variable region (VL) comprising a CDRL1 domain comprising the sequence described in SEQ ID NO: 16 or 17, a CDRL2 domain comprising the sequence described in SEQ ID NO: 18 or 19, and a CDRL3 domain comprising the sequence described in SEQ ID NO: 20.

[0067] In embodiment A, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising a heavy chain variable region (VH) comprising a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, 11, 12, 13, or 14, and a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and a light chain variable region (VL) comprising a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16 or 17, a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18 or 19, and a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0068] One embodiment of A is an isolated antibody or an antigen-binding fragment thereof, which comprises (a) a heavy chain variable region (VH) comprising i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0069] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a)i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b)i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0070] A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0071] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 10, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0072] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a)i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b)i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0073] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0074] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain containing the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain containing the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain containing the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain containing the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain containing the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain containing the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0075] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 11, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0076] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a)i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b)i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0077] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0078] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a)i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b)i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0079] A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 12, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0080] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0081] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0082] A further embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 13, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0083] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 16, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0084] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 18, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0085] Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) comprising (a) i) a CDRH1 domain comprising the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain comprising the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain comprising the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) comprising i) a CDRL1 domain comprising the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain comprising the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain comprising the sequence described in SEQ ID NO: 20. Another embodiment of Embodiment A is an isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (VH) including (a) i) a CDRH1 domain consisting of the sequence described in SEQ ID NO: 9, ii) a CDRH2 domain consisting of the sequence described in SEQ ID NO: 14, and iii) a CDRH3 domain consisting of the sequence described in SEQ ID NO: 15, and (b) a light chain variable region (VL) including i) a CDRL1 domain consisting of the sequence described in SEQ ID NO: 17, ii) a CDRL2 domain consisting of the sequence described in SEQ ID NO: 19, and iii) a CDRL3 domain consisting of the sequence described in SEQ ID NO: 20.

[0086] In embodiment B, the disclosure provides an isolated antibody or antigen-binding fragment thereof that specifically binds to CD40L, comprising (a) a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1, 2, 3, or 4, and (b) a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5, 6, 7, or 8.

[0087] Embodiment B is an isolated antibody or its antigen-binding fragment, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5.

[0088] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 6.

[0089] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 7.

[0090] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 8.

[0091] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5.

[0092] A further embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 6.

[0093] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 7.

[0094] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 1 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 8.

[0095] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 3 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5.

[0096] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 3 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 6.

[0097] Another embodiment of Embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 3 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 7.

[0098] A further embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 3 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 8.

[0099] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 4 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 5.

[0100] A further embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 4 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 6.

[0101] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 4 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 7.

[0102] Another embodiment of embodiment B is an isolated antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence described in SEQ ID NO: 4 and a light chain variable region (VL) having the amino acid sequence described in SEQ ID NO: 8.

[0103] Embodiment C provides an isolated antibody of any embodiment described in this application, wherein such antibody comprises an Fc region, the Fc region being manipulated to suppress or eliminate one or more Fc effector functions.

[0104] One embodiment of Embodiment C is one in which the Fc region is manipulated to suppress or eliminate binding to an Fc receptor (FcR) or to C1q. In a particular embodiment of Embodiment C, the FcR is FCγRIa (CD64), FCγRIIa (CD32), FCγRIIIa (CD16a), or FCγRIIIb (CD16b). In another embodiment of Embodiment C, the FcR is FcγRIIa.

[0105] Embodiment C is an isolated antibody according to any one embodiment of the present invention, wherein such antibody is an isotype of IgG1, IgG2, IgG3, or IgG4, or any combination thereof or a hybrid. Another embodiment of Embodiment C is an isolated antibody according to any one embodiment of the present invention, wherein such antibody is an IgG1 isotype, and the heavy chain constant region contains the amino acid sequence described in SEQ ID NO: 21 (P238S and N297G). As shown in Experiment 3, none of the antibody clones provided in this disclosure, manipulated using the Fc of SEQ ID NO: 21, bound to FCγRIa, FCγRIIa, FCγRIIIa, or FCγRIIIb. As shown in Experiment 4, none of the antibody clones provided in this disclosure, manipulated using the Fc of SEQ ID NO: 21, bound to C1q.

[0106] A mutation at position 297 of the IgG backbone Fc (N297A) has been shown to inhibit IgG glycosylation and Fc gamma receptor binding. A mutation at position 265 of the IgG backbone Fc (D265A) has been shown to inhibit Fc gamma receptor binding. Accordingly, embodiments of the present invention containing these mutations in the Fc region have been prepared. One embodiment of Embodiment C is an isolated antibody according to any one of the embodiments of the present disclosure (D265A and N297A), comprising a heavy chain constant region containing the amino acid sequence described in SEQ ID NO: 22.

[0107] U.S. Patent Application Publication No. 2014 / 0294834 discloses a mutation in the heavy chain constant region. Therefore, another embodiment is an isolated antibody by any one of the embodiments having the Fc region disclosed in this application, comprising a heavy chain constant region having the amino acid sequence described in SEQ ID NO: 23 (T299K-MM131).

[0108] Another embodiment is an isolated antibody according to any one of the embodiments having an Fc region disclosed in this application, comprising a heavy chain constant region containing the amino acid sequence described in SEQ ID NO: 24 (N299K).

[0109] In certain embodiments, the isolated antibody is a heavy chain variable region connected to a heavy chain constant region. In one embodiment, the heavy chain variable region VH1 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 42. In another embodiment, the heavy chain variable region VH2 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 43. In yet another embodiment, the heavy chain variable region VH3 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 44. In yet another embodiment, the heavy chain variable region VH4 is directly connected to a heavy chain constant region having P238S and N297G, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 45.

[0110] In one embodiment, the heavy chain variable region VH1 is directly connected to the heavy chain constant region having D265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 46. In another embodiment, the heavy chain variable region VH2 is directly connected to the heavy chain constant region having D265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 47. In yet another embodiment, the heavy chain variable region VH3 is directly connected to the heavy chain constant region having 265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 48. In yet another embodiment, the heavy chain variable region V4 is directly connected to the heavy chain constant region having 265A / N297A, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 49.

[0111] In yet another embodiment, the heavy chain variable region VH1 is directly connected to the heavy chain constant region having an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 50. In yet another embodiment, the heavy chain variable region VH2 is directly connected to the heavy chain constant region having an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 51. In yet another embodiment, the heavy chain variable region VH3 is directly connected to the heavy chain constant region having an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 51. In yet another embodiment, the heavy chain variable region VH4 is directly connected to the heavy chain constant region having an IgG1 / IgG4 hybrid, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 53.

[0112] In another embodiment, the heavy chain variable region VH1 is directly connected to the heavy chain constant region having IgG4, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 54. In yet another embodiment, the heavy chain variable region VH2 is directly connected to the heavy chain constant region having IgG4, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 55. In yet another embodiment, the heavy chain variable region VH3 is directly connected to the heavy chain constant region having IgG4, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 56. In yet another embodiment, the heavy chain variable region VH4 is directly connected to the heavy chain constant region having IgG4, and the isolated antibody heavy chain consists of the amino acid sequence described in SEQ ID NO: 57.

[0113] Another embodiment is an isolated antibody from any one embodiment having a variable light chain and a light chain constant region containing the sequence described in SEQ ID NO: 25. Another embodiment is a variable light chain 1 having a light chain constant region, the amino acid sequence of which is described in SEQ ID NO: 38. Another embodiment is a variable light chain 2 having a light chain constant region, the amino acid sequence of which is described in SEQ ID NO: 39. Yet another embodiment is a variable light chain 3 having a light chain constant region, the amino acid sequence of which is described in SEQ ID NO: 40. Yet another embodiment is a variable light chain 4 having a light chain constant region, the amino acid sequence of which is described in SEQ ID NO: 41.

[0114] Another embodiment is a method for treating a subject having a disease or disorder related to CD40L, comprising administering to such subject a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0115] Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 42 and the light chain sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 43 and the light chain sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 44 and the light chain sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 45 and the light chain sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 42 and the light chain sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 43 and the light chain sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 44 and the light chain sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 45 and the light chain sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 42 and the light chain sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 43 and the light chain sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 44 and the light chain sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 45 and the light chain sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 42 and the light chain sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 43 and the light chain sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 44 and the light chain sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 45 and the light chain sequence described in SEQ ID NO: 41.

[0116] Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 46 and the light chain of the amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 47 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 48 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 49 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 46 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 47 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 48 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 49 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 46 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 47 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 48 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 49 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 46 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 47 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 48 and the light chain amino acid sequence described in SEQ ID NO: 41.Another embodiment is an isolated antibody having the heavy chain amino acid sequence described in SEQ ID NO: 49 and the light chain amino acid sequence described in SEQ ID NO: 41.

[0117] Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 50 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 51 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 52 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 53 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 50 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 51 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 52 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 53 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 50 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 51 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 52 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 53 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 50 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 51 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 52 and the light chain amino acid sequence described in SEQ ID NO: 41.Another embodiment is an isolated antibody having the heavy chain amino acid sequence described in SEQ ID NO: 53 and the light chain amino acid sequence described in SEQ ID NO: 41.

[0118] Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 54 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 55 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 56 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 57 and the light chain amino acid sequence described in SEQ ID NO: 38. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 54 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 55 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 56 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 57 and the light chain amino acid sequence described in SEQ ID NO: 39. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 54 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 55 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 56 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 57 and the light chain amino acid sequence described in SEQ ID NO: 40. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 54 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 55 and the light chain amino acid sequence described in SEQ ID NO: 41. Another embodiment is an isolated antibody having the heavy chain of the amino acid sequence described in SEQ ID NO: 56 and the light chain amino acid sequence described in SEQ ID NO: 41.Another embodiment is an isolated antibody having the heavy chain amino acid sequence described in SEQ ID NO: 57 and the light chain amino acid sequence described in SEQ ID NO: 41.

[0119] Another embodiment is a method for treating a subject having a neurodegenerative or neuromuscular disease or disorder, an inflammatory or immune disease or disorder, or an autoimmune disease, comprising administering to such subject a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0120] Another embodiment is a method for treating a subject having an autoimmune disease selected from the group consisting of systemic lupus erythematosus, type 1 diabetes mellitus, myasthenia gravis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, inflammatory bowel disease, immune thrombocytopenic purpura, Graves' disease, and rheumatoid arthritis, comprising administering to such subject a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0121] Another embodiment is a method for treating subjects having a neurodegenerative disease or neuromuscular disorder selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multifocal motor neuropathy, primary lateral sclerosis, spinal muscular atrophy, Kennedy disease, and spinocerebellar ataxia, comprising administering to such subjects a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0122] Another embodiment is a method for treating a subject having amyotrophic lateral sclerosis, comprising administering to such subject a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0123] Another embodiment is a method for inhibiting an immune response in a subject, which includes administering to such subject a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein.

[0124] Another embodiment is a method for inhibiting an immune response in a subject, comprising administering to such subject a therapeutically effective amount of an antibody or an antigen-binding fragment thereof, wherein the immune response is graft-versus-host disease or organ transplant rejection.

[0125] Another embodiment is a method according to any of the methods of the present disclosure, in which the antibody or antibody fragment is administered in combination with another therapeutic agent. One embodiment is a method according to any of the methods of the present disclosure, in which the antibody or antibody fragment is administered in combination with a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0126] Another embodiment is a method according to any of the methods of the present disclosure, in which the antibody or antibody fragment is administered in combination with a CTLA4-Ig fusion protein, which is a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0127] Another embodiment is a method according to any of the methods of the present disclosure, in which the antibody or antibody fragment is administered in combination with abatacept, beratacept, or galiximab, which is a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Examples]

[0128] Example 1: CD40L binding assay

[0129] To compare the CD40L binding of all 16 antibody clones with that of 5c8 or AT-1501, binding assays were performed using two clones together with 5c8 and AT-1501 on the same 96-well assay plate. The binding levels of the antibodies disclosed herein, compared to reference antibodies 5c8-19 and AT1501, were determined using a three-part sandwich ELISA assay. 96-well polystyrene plates were coated with 2 ug / ml recombinant human CD40L (BioLegend catalog no. 591706) in PBS, and 50 ul / well was added to a Costar high-binding 1 / 2 area 96-well assay plate (Corning 3690). The plates were incubated overnight at 4°C. To prevent background binding, the plates were blocked at room temperature for 1 hour using (1×) PBS / 1.0% BSA (140 ul / well). Add binding curve 5C8 or AT1501 (gradually 2-fold dilution from 2 ug / ml) (50 ul / well) and incubate at room temperature for 1 hour. Wash the plate and incubate at room temperature for 1 hour with 1:10,000 dilution of HRP-(Fab2) donkey anti-human IgG (Fc specific) (Jackson Immuno.709-036-098) (50 ul / well). Wash the plate and add TMB substrate (Surmodics BioFX TMBW-1000-01) (50 ul / well). After 5 minutes at room temperature, stop color development with 25 ul of 2NH2SO4. Read the plate using a Molecular Devices SpectraMax M5 microplate reader with the SoftMax Pro 6.2.2 program and measure the absorbance at 450 nm.

[0130] The relative CD40L binding capacity was calculated as follows: [IC50 (clone) / IC50 (5c8 or AT-1501) × 100%]. The titer ranking of the 16 clones relative to 5c8 is shown in Figure 1A, and the titer ranking relative to AT-1501 is shown in Figure 1B. The resulting binding curves are shown in Figures 2A-2Q (comparison between the 16 clones and 5c8, with Figure 2H showing a comparison between AT-1501 and 5c8), and Figures 3A-3Q (comparison between the 16 clones and AT-1501, with Figure 3H showing a comparison between AT-1501 and 5c8). The IC50, LCL, UCL, and relative titer of each clone are shown in Table 2 (comparison with 5c8) and Table 3 (comparison with AT-1501). [Table 2-1] [Table 2-2] [Table 3-1] [Table 3-2]

[0131] Example 2: Binding activity to human FcγRI, FcγRIIa, FcγRIIIa, and FcγRIIIb

[0132] Sixteen VH / VL antibody clones were constructed using IgG1 Fc (SEQ ID NO: 21) containing two mutations, P238S and N297G. These antibody clones were assayed for their Fc effector function in binding to human FcγRI, FcγRIIa, and FcγRIIIa.

[0133] Anti-CD40L antibody (including abatacept as a negative control) was diluted to 2 ug / ml in (1×) PBS, and 50 ul / well was added to a Costar high-binding 1 / 2 area 96-well assay plate (Corning 3690), and incubated overnight at 4°C. To prevent background binding, the plate was blocked with (1×) PBS / 1.0% BSA (140 ul / well) at room temperature for 1 hour. Recombinant human FcγRIs IIa, IIIa, and IIIb (gradually diluted 2-fold from 5 ug / ml) were added (50 ul / well), and incubated at room temperature for 1 hour. The plates were washed and incubated with mouse anti-human CD16 (anti-FcRIII), CD32 (anti-FcRIIa), or CD64 (anti-FcRI) (eBioSciences / Invitrogen 14-0168-82, 16-0329-81, 14-0649-82) at 2 ug / ml (50 ul / well) at room temperature for 1 hour. The plates were washed and incubated with HRP-(Fab2) goat anti-mouse IgG (Fc-specific) at a dilution of 1:10,000 (Jackson Immuno. 116-036-071) at room temperature for 1 hour (50 ul / well). The plates were washed and TMB substrate (Surmodics BioFX TMBW-1000-01) was added (50 ul / well). After 5 minutes at room temperature, color development was stopped with 2NH2SO4 (25 ul / well). The plates were read using a Molecular Devices SpectraMax M5 microplate reader with the SoftMax Pro 6.2.2 program, and the absorbance at 450 nm was measured.

[0134] As can be seen from the figures, the clones disclosed in this application were negative for binding to three Fc receptors, with 5c8 binding to FcγRI and FcγRIIa, but not to FcγRIIIa or FcγRIIIb. Figures 4A and 4B each show the binding of eight antibody clones to FcγRIa compared to the binding curves of 5c8, AT-1501, and abatacept. Figures 5A and 5B each show the binding of eight antibody clones to FcγRIIa compared to the binding curves of 5c8, AT-1501, and abatacept. Figures 6A and 6B each show the binding of eight antibody clones to FcγRIIIa compared to the binding curves of 5c8, AT-1501, and abatacept. Figures 6C and 6D each show the binding of eight antibody clones to FcγRIIIb compared to the binding curves of 5c8, AT-1501, and abatacept. (Figures 4A, 5A, 6A, and 6C show the results using antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4, while Figures 4B, 5B, 6B, and 6D show the results using antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2).

[0135] Example 4 Binding activity to C1q

[0136] Anti-CD40L antibody (including abatacept as a negative control) was diluted to 2 ug / ml in (1×) PBS, and 50 ul / well was added to a Costar high-binding 1 / 2 area 96-well assay plate (Corning 3690). The plate was incubated overnight at 4°C. To prevent background binding, the plate was blocked with (1×) PBS / 1.0% BSA (140 ul / well) at room temperature for 1 hour. Natural human C1q protein (Abcam ab96363) with binding curves was added in stepwise 2-fold dilutions starting from 10 ug / ml (50 ul / well) and incubated at room temperature for 1 hour. After washing the plate, HRP-sheep anti-human C1q (Abcam ab46191) at a dilution of 1:400 was added (50 ul / well) and incubated at room temperature for 1 hour. After washing the plates, TMB substrate (Surmodics BioFX TMBW-1000-01) was added (50 μl / well). After 5 minutes at room temperature, color development was stopped with (25 μl / well) 2NH2SO4. The plates were read using a Molecular Devices SpectraMax M5 microplate reader with the SoftMax Pro 6.2.2 program, and the absorbance at 450 nm was measured.

[0137] As can be seen from the figures, all antibodies obtained from the 16 antibody clones disclosed in this application were negative for binding to C1q, except for 5c8, which showed significant binding. Figures 7A and 7B show the binding of eight antibody clones to C1q compared to the binding curves of 5c8, AT-1501, and abatacept, respectively. Figure 7A shows the results for antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4, and Figure 7B shows the results for antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2.

Claims

1. (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain comprising the sequence described in Sequence ID No. 10, 11, 12, 13, or 14, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain comprising the sequence described in Sequence ID No. 16 or 17, ii) A CDRL2 domain comprising the sequence described in Sequence ID No. 18 or 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) An isolated antibody or its antigen-binding fragment that specifically binds to CD40L, including [the specified substance].

2. (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 10, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 10, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 10, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 10, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and comprising a heavy chain variable region (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 11, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 11, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 11, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 11, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 12, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 12, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 12, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 12, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 13, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 13, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 13, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 13, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 13, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 14, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 14, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 18, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 14, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 16, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) Includes, or (a) i) A CDRH1 domain containing the sequence described in Sequence ID No. 9, ii) A CDRH2 domain containing the sequence described in Sequence ID No. 14, iii) CDRH3 domain containing the sequence described in Sequence ID No. 15 and Heavy chain variable region including (V Ｈ ), and (b) i) A CDRL1 domain containing the sequence described in Sequence ID No. 17, ii) A CDRL2 domain containing the sequence described in Sequence ID No. 19, iii) CDRL3 domain containing the sequence described in Sequence ID No. 20 and Light chain variable region including (V Ｌ ) The isolated antibody or its antigen-binding fragment according to claim 1, comprising:

3. (a) Heavy chain variable region having the amino acid sequence described in SEQ ID NOs: 1, 2, 3, or 4 (V Ｈ ) and (b) Light chain variable region having the amino acid sequence described in SEQ ID NO: 5, 6, 7, or 8 (V Ｌ ) An isolated antibody or its antigen-binding fragment that specifically binds to CD40L, including [the specified substance].

4. (a) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 5 (V Ｌ ),or (b) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 6 (V Ｌ ),or (c) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 7 (V Ｌ ),or (d) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 8 (V Ｌ ),or (e) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 5 (V Ｌ ),or (f) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 6 (V Ｌ ),or (g) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 7 (V Ｌ ),or (h) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 1 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 8 (V Ｌ ),or (i) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 3 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 5 (V Ｌ ),or (j) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 3 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 6 (V Ｌ ),or (k) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 3 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 7 (V Ｌ ),or (l) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 3 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 8 (V Ｌ ),or (m) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 4 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 5 (V Ｌ ),or (n) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 4 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 6 (V Ｌ ),or (o) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 4 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 7 (V Ｌ ),or (p) Heavy chain variable region having the amino acid sequence described in Sequence ID No. 4 (V Ｈ ) and a light chain variable region having the amino acid sequence described in Sequence ID No. 8 (V Ｌ ) The isolated antibody or its antigen-binding fragment according to claim 3, comprising:

5. The isolated antibody according to any one of claims 1 to 4, wherein the antibody or antigen-binding fragment comprises an Fc region, and the Fc region is manipulated to suppress or eliminate one or more Fc effector functions.

6. The isolated antibody according to claim 5, wherein one or more Fc effector functions are FcR binding or complement binding.

7. The isolated antibody according to any one of claims 1 to 6, wherein the antibody is of the IgG1 isotype and the heavy chain constant region comprises the amino acid sequence described in SEQ ID NO:

21.

8. An isolated antibody according to any one of claims 1 to 6, further comprising a heavy chain constant region containing the amino acid sequence described in SEQ ID NO:

22.

9. The isolated antibody according to any one of claims 1 to 6, wherein the chain constant region comprises the amino acid sequence described in SEQ ID NO:

23.

10. An isolated antibody according to any one of claims 1 to 6, further comprising a heavy chain constant region containing the amino acid sequence described in SEQ ID NO:

24.

11. The isolated antibody according to any one of claims 1 to 10, further comprising a light chain constant region having the amino acid sequence described in SEQ ID NO:

25.

12. The aforementioned antibody is (a) A heavy chain having the amino acid sequence described in SEQ ID NO: 42 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (b) A heavy chain having the amino acid sequence described in SEQ ID NO: 42 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (c) A heavy chain having the amino acid sequence described in SEQ ID NO: 42 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (d) A heavy chain having the amino acid sequence described in SEQ ID NO: 42 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (e) A heavy chain having the amino acid sequence described in SEQ ID NO: 43 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (f) A heavy chain having the amino acid sequence described in SEQ ID NO: 43 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (g) A heavy chain having the amino acid sequence described in SEQ ID NO: 43 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (h) A heavy chain having the amino acid sequence described in SEQ ID NO: 43 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (i) A heavy chain having the amino acid sequence described in SEQ ID NO: 44 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (j) A heavy chain having the amino acid sequence described in SEQ ID NO: 44 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (k) A heavy chain having the amino acid sequence described in SEQ ID NO: 44 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (l) A heavy chain having the amino acid sequence described in SEQ ID NO: 44 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (m) A heavy chain having the amino acid sequence described in SEQ ID NO: 45 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (n) A heavy chain having the amino acid sequence described in SEQ ID NO: 45 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (o) A heavy chain having the amino acid sequence described in SEQ ID NO: 45 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (p) Heavy chain having the amino acid sequence described in SEQ ID NO: 45 and light chain having the amino acid sequence described in SEQ ID NO: 41 The isolated antibody, including the following.

13. The aforementioned antibody is (a) A heavy chain having the amino acid sequence described in SEQ ID NO: 46 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (b) A heavy chain having the amino acid sequence described in SEQ ID NO: 46 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (c) A heavy chain having the amino acid sequence described in SEQ ID NO: 46 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (d) A heavy chain having the amino acid sequence described in SEQ ID NO: 46 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (e) A heavy chain having the amino acid sequence described in SEQ ID NO: 47 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (f) A heavy chain having the amino acid sequence described in SEQ ID NO: 47 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (g) A heavy chain having the amino acid sequence described in SEQ ID NO: 47 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (h) A heavy chain having the amino acid sequence described in SEQ ID NO: 47 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (i) A heavy chain having the amino acid sequence described in SEQ ID NO: 48 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (j) A heavy chain having the amino acid sequence described in SEQ ID NO: 48 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (k) A heavy chain having the amino acid sequence described in SEQ ID NO: 48 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (l) A heavy chain having the amino acid sequence described in SEQ ID NO: 48 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (m) A heavy chain having the amino acid sequence described in SEQ ID NO: 49 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (n) A heavy chain having the amino acid sequence described in SEQ ID NO: 49 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (o) A heavy chain having the amino acid sequence described in SEQ ID NO: 49 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (p) Heavy chain having the amino acid sequence described in SEQ ID NO: 49 and light chain having the amino acid sequence described in SEQ ID NO: 41 The isolated antibody, including the following.

14. The aforementioned antibody is (a) A heavy chain having the amino acid sequence described in SEQ ID NO: 50 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (b) A heavy chain having the amino acid sequence described in SEQ ID NO: 50 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (c) A heavy chain having the amino acid sequence described in SEQ ID NO: 50 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (d) A heavy chain having the amino acid sequence described in SEQ ID NO: 50 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (e) A heavy chain having the amino acid sequence described in SEQ ID NO: 51 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (f) A heavy chain having the amino acid sequence described in SEQ ID NO: 51 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (g) A heavy chain having the amino acid sequence described in SEQ ID NO: 51 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (h) A heavy chain having the amino acid sequence described in SEQ ID NO: 51 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (i) A heavy chain having the amino acid sequence described in SEQ ID NO: 52 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (j) A heavy chain having the amino acid sequence described in SEQ ID NO: 52 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (k) A heavy chain having the amino acid sequence described in SEQ ID NO: 52 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (l) A heavy chain having the amino acid sequence described in SEQ ID NO: 52 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (m) A heavy chain having the amino acid sequence described in SEQ ID NO: 53 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (n) A heavy chain having the amino acid sequence described in SEQ ID NO: 53 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (o) A heavy chain having the amino acid sequence described in SEQ ID NO: 53 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (p) Heavy chain having the amino acid sequence described in SEQ ID NO: 53 and light chain having the amino acid sequence described in SEQ ID NO: 41 The isolated antibody, including the following.

15. The aforementioned antibody is (a) A heavy chain having the amino acid sequence described in SEQ ID NO: 54 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (b) A heavy chain having the amino acid sequence described in SEQ ID NO: 54 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (c) A heavy chain having the amino acid sequence described in SEQ ID NO: 54 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (d) A heavy chain having the amino acid sequence described in SEQ ID NO: 54 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (e) A heavy chain having the amino acid sequence described in SEQ ID NO: 55 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (f) A heavy chain having the amino acid sequence described in SEQ ID NO: 55 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (g) A heavy chain having the amino acid sequence described in SEQ ID NO: 55 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (h) A heavy chain having the amino acid sequence described in SEQ ID NO: 55 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (i) A heavy chain having the amino acid sequence described in SEQ ID NO: 56 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (j) A heavy chain having the amino acid sequence described in SEQ ID NO: 56 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (k) A heavy chain having the amino acid sequence described in SEQ ID NO: 56 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (l) A heavy chain having the amino acid sequence described in SEQ ID NO: 56 and a light chain having the amino acid sequence described in SEQ ID NO: 41, or (m) A heavy chain having the amino acid sequence described in SEQ ID NO: 57 and a light chain having the amino acid sequence described in SEQ ID NO: 38, or (n) A heavy chain having the amino acid sequence described in SEQ ID NO: 57 and a light chain having the amino acid sequence described in SEQ ID NO: 39, or (o) A heavy chain having the amino acid sequence described in SEQ ID NO: 57 and a light chain having the amino acid sequence described in SEQ ID NO: 40, or (p) Heavy chain having the amino acid sequence described in SEQ ID NO: 57 and light chain having the amino acid sequence described in SEQ ID NO: 41 The isolated antibody, including the following.

16. A method for treating a subject having a disease or disorder related to CD40L, The method according to any one of claims 1 to 15, comprising administering a therapeutically effective amount of antibody or antibody fragment to the subject.

17. A method for treating a subject of neurodegenerative or neuromuscular disease or disorder, inflammatory or immune disease or disorder, or autoimmune disease, The method according to any one of claims 1 to 15, comprising administering a therapeutically effective amount of antibody or antibody fragment to the subject.

18. The method according to claim 17, wherein the disease or disorder is an autoimmune disease selected from the group consisting of systemic lupus erythematosus, type 1 diabetes mellitus, myasthenia gravis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, inflammatory bowel disease, immune thrombocytopenic purpura, Graves' disease, and rheumatoid arthritis.

19. The method according to claim 17, wherein the disease or disorder is a neurodegenerative disease or neuromuscular disorder selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multifocal motor neuropathy, primary lateral sclerosis, spinal muscular atrophy, Kennedy disease, or spinocerebellar ataxia.

20. The method according to claim 17, wherein the disease or disorder is amyotrophic lateral sclerosis.

21. A method for inhibiting the immune response in a target, The method according to any one of claims 1 to 15, comprising administering a therapeutically effective amount of antibody or antibody fragment to the subject.

22. The method according to claim 17, wherein the immune response is graft-versus-host disease or organ transplant rejection.

23. The method according to any one of claims 16 to 22, wherein the antibody or antibody fragment is administered in combination with another therapeutic agent.

24. The method according to claim 23, wherein the antibody is administered in combination with a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

25. The method according to claim 24, wherein the compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80 is a CTLA4-Ig fusion protein, or the compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80 is abatacept, beratacept, or galiximab.

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