Methods for treating kidney diseases or disorders

Abstract

Provided herein are methods and kits for treating kidney-related disease disorders and complications, such as Burger's disease and renal transplant rejection. Also provided are methods of predicting or measuring response to treatment by measuring the level of a biomarker in a sample, as well as methods of modulating the level of a biomarker.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 365,448, filed on May 27, 2022, which is incorporated by reference in its entirety.

[0003] References to sequence listings

[0004] This application is submitted with a sequence listing in electronic format. The sequence listing is provided as a file titled "ELDN.011WO.xml" created on May 1, 2023, which is 27,088 bytes in size. The information in the sequence listing in electronic format is incorporated herein by reference in its entirety.

[0005] background

[0006] field

[0007] The present disclosure relates to methods for treating an autoimmune disorder in a subject, including, for example, a kidney-related autoimmune disorder, such as a kidney-related disease, disorder or complication, such as Buerger's disease or a kidney transplant. Methods for predicting and / or measuring a response to treatment are also provided, including measuring the level of one or more biomarkers. Embodiments also relate to methods for regulating the level of one or more biomarkers.

[0008] Related technical notes

[0009] The kidney is an essential organ for filtering a subject's blood and regulating mineral content. Many diseases and disorders are associated with the kidney, including chronic kidney disease (CKD), diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infections, nephrolithiasis, and IgA nephropathy.

[0010] IgA nephropathy (also known as Buerger's disease or IgAN) is a type of chronic kidney disease. Over approximately 10-20 years, IgA accumulates in the subject's glomeruli and causes the filtration system in the kidney to become blocked. This in turn leads to activation of the immune system and general inflammation around the glomeruli of the kidney. Aggressive IgAN may also affect other major organs, such as the liver, skin, and cardiovascular system.

[0011] Severe kidney disease in an individual may result in the need for a kidney organ transplant. However, an individual typically must receive a donated kidney from a donor with ABO blood type and crossmatch (human leukocyte antigen-HLA) compatibility. Even if a suitable kidney is successfully transplanted, the individual must continue to take immunosuppressants for the rest of their lives. Otherwise, the risk of their immune rejection of the new organ is high. Another potential complication after a kidney transplant is graft-versus-host disease, in which the donor's leukocytes that remain in the newly transplanted kidney recognize their new host as foreign and attack the individual's somatic cells. The high risk of complications after a kidney transplant remains a major obstacle in the medical field.

[0012] Overview

[0013] The present application discloses a method for predicting the response of a subject with a kidney-related disease or disorder to treatment with a compound for CD40L or CD40. In some embodiments, the method includes collecting a sample from the subject; and measuring the concentration of at least one biomarker present in the sample, wherein the at least one biomarker is CXCL9, CXCL10, IgM, C3, IgA, IgE, CD40L and CD40; wherein the concentration of the at least one biomarker predicts that the subject may respond to treatment of the kidney-related disease or disorder with a compound for CD40L or CD40. In some embodiments, the kidney-related disease or disorder includes inflammation. In some embodiments, the subject is a mammal and / or a human. In some embodiments, the compound blocks the interaction between CD40 and CD40L. In some embodiments, the compound is an anti-CD40L antibody or an anti-CD40 antibody. In some embodiments, the compound is AT-1501. In some embodiments, the compound is MR1 ​​or 5c8. In some embodiments, the kidney-related disease or disorder is selected from the group consisting of: colitis, drug-induced lupus nephritis, lupus nephritis, nephropathy, chronic kidney disease, graft-versus-host disease, immune graft response, kidney transplantation, transplant rejection, atherosclerosis, diabetic kidney disease, acute renal injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis and IgA nephropathy. In some embodiments, the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia or Graves' disease. In some embodiments, the sample is blood, urine, saliva or mucus. In some embodiments, the sample is urine.

[0014] Also disclosed herein is a method for treating a subject with a kidney-related disease or disorder. In some embodiments, the method includes administering a therapeutically effective dose of a compound for CD40L or CD40. In some embodiments, the kidney-related disease or disorder includes inflammation. In some embodiments, the subject is a mammal and / or a human. In some embodiments, the compound blocks the interaction between CD40 and CD40L. In some embodiments, the compound is an anti-CD40L antibody or an anti-CD40 antibody. In some embodiments, the compound is AT-1501. In some embodiments, the compound is MR1 ​​or 5c8. In some embodiments, the compound is administered at least once every three weeks. In some embodiments, the compound is administered at a dose between 200 mg / kg and 100 mg / kg, preferably at a dose between 1 mg / kg and 20 mg / kg. In some embodiments, the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, nephropathy, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute renal injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis or IgA nephropathy. In some embodiments, the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia or Graves' disease. In some embodiments, the method further comprises administering a second pharmaceutically effective compound. In some embodiments, the second compound blocks the interaction between CD28 and CD86 or between CD28 and CD80. In some embodiments, the second compound targets at least one of the biomarkers selected from the group consisting of: interleukins, cytokines, and pro-inflammatory markers. In some embodiments, the second compound is CTLA4-Ig fusion protein, abatacept, belatacept, or galiximab. In some embodiments, the compound is administered orally, parenterally, or topically. In some embodiments, the compound is administered parenterally. In some embodiments, the compound is administered by injection, most preferably intravenously, intramuscularly, or subcutaneously. In some embodiments, the compound targeting CD40L or CD40 is an antibody comprising: a heavy chain variable region (V H), the heavy chain variable region comprises: (1) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9; (2) a CDRH2 domain comprising the sequence shown in SEQ ID NO:10, 11, 12, 13 or 14; and (3) a CDRH3 domain comprising the sequence shown in SEQ ID NO:15; and further comprises a light chain variable region (V L ), the light chain variable region comprises: (4) a CDRL1 domain comprising the sequence shown in SEQ ID NO: 16 or 17; (5) a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and (6) a CDRL3 domain comprising the sequence shown in SEQ ID NO: 20. In some embodiments, the compound targeting CD40L or CD40 is an antibody comprising: a heavy chain variable region (V H ), the heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 1, 2, 3 or 4; and the light chain variable region (V L ), the light chain variable region having an amino acid sequence as shown in SEQ ID NO:5, 6, 7 or 8.

[0015] Also disclosed herein is a method for regulating the concentration of at least one biomarker in a subject. In some embodiments, the method includes administering to the subject a therapeutically effective amount of a compound for CD40L or CD40, wherein the biomarker is selected from the group consisting of: IgA, IgE, CD40L, CD40, C3, CXCL9, CXCL10, and IgM. In some embodiments, the concentration of at least one biomarker increases after administration of the compound. In some embodiments, the concentration of at least one biomarker decreases after administration of the compound. In some embodiments, after administration of the compound, the concentration of at least one biomarker increases, and the concentration of at least one other biomarker decreases. In some embodiments, the subject suffers from a kidney-related disease or disorder. In some embodiments, the kidney-related disease or disorder includes inflammation. In some embodiments, the subject is a mammal and / or a human. In some embodiments, the compound blocks the interaction between CD40 and CD40L. In some embodiments, the compound is an anti-CD40L antibody or an anti-CD40 antibody. In some embodiments, the compound is AT-1501. In some embodiments, the compound is MR1 ​​or 5c8. In some embodiments, the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, nephropathy, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute renal injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis or IgA nephropathy. In some embodiments, the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia or Graves' disease. In some embodiments, the compound is administered at least once every three weeks. In some embodiments, the compound is administered at a dose between 200 mg / kg and 100 mg / kg, preferably at a dose between 1 mg / kg and 20 mg / kg. In some embodiments, the compound is administered orally, parenterally, or topically. In some embodiments, the compound is administered parenterally. In some embodiments, the compound is administered by injection, most preferably intravenous, intramuscular, or subcutaneous injection. In some embodiments, the compound targeting CD40L or CD40 is an antibody comprising a heavy chain variable region (V H), the heavy chain variable region comprises: (1) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9; (2) a CDRH2 domain comprising the sequence shown in SEQ ID NO:10, 11, 12, 13 or 14; and (3) a CDRH3 domain comprising the sequence shown in SEQ ID NO:15; and comprises a light chain variable region (V L ), the light chain variable region comprises: (4) a CDRL1 domain comprising the sequence shown in SEQ ID NO: 16 or 17; (5) a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and (6) a CDRL3 domain comprising the sequence shown in SEQ ID NO: 20. In some embodiments, the compound targeting CD40L or CD40 is an antibody comprising: a heavy chain variable region (V H ), the heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 1, 2, 3 or 4; and the light chain variable region (V L ), the light chain variable region having an amino acid sequence as shown in SEQ ID NO:5, 6, 7 or 8. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1A MR1 concentration over time is shown using a linear concentration (Y) axis.

[0017] Figure 1B MR1 concentration over time is shown using the log concentration (Y) axis.

[0018] Figure 2 is a graph of the average daily body weight measurements for the control and treatment groups starting on day 40 until peak body weight was reached.

[0019] Figure 3 Mean daily body weight measurements from peak body weight until death are depicted for the control and treatment groups.

[0020] Figure 4 Kaplan-Meier survival plots for the control and treatment groups as calculated by the Cox proportional hazards model are shown.

[0021] Figure 5A is a graph of average daily body weight measurements for the control and MR1 treated groups starting on day 40 until peak body weight was reached.

[0022] Figure 5B is a graph depicting the mean daily body weight measurements from peak body weight until death for the control and MR1 treated groups.

[0023] Figure 5CShown is the time to disease onset based on the time to progression to a neurological function score of 2 in the control and MR1-treated groups.

[0024] Figure 5D Mean daily body weight measurements from peak body weight until death are shown for the control and MR1 treated groups.

[0025] Figure 6 is a graph showing the relative binding of JB5 antibody (circles, dashed line), hu5c8 antibody (squares-solid line) and control CTLA4-IgG1 (triangles) to human CD40L.

[0026] Figure 7 Provided are the amino acid sequences of hu5c8 and JB5 VL region (SEQ ID NO: 1), hu5c8 and JB5 VH region (SEQ ID NO: 2), Fc region (SEQ ID NO: 3), JB5 Fc region (SEQ ID NO: 4), JB5-R28K VL region (SEQ ID NO: 5), JB5-K74R VH region (SEQ ID NO: 6), and the JB5 light chain amino acid sequence (SEQ ID NO: 7).

[0027] Figure 8 Provided is a synthetic nucleotide sequence encoding the light chain of the anti-CD40L antibody JB5 (SEQ ID NO: 8), with capital letters representing exons of the synthetic gene and lowercase letters representing intron sequences of the synthetic gene; and also provided is a heavy chain amino acid sequence of the anti-CD40L antibody JB5 (SEQ ID NO: 9).

[0028] Fig. 9 A synthetic nucleic acid sequence encoding the heavy chain of the anti-CD40L antibody JB5 is provided (SEQ ID NO: 10), with capital letters representing exons of the synthetic gene and lowercase letters representing intronic sequences of the synthetic gene.

[0029] Fig.10 Provided are the amino acid sequence of the anti-CD40L antibody JB5-R28K (SEQ ID NO: 11); a synthetic nucleic acid sequence encoding the light chain of the anti-CD40L antibody JB5-R28K (SEQ ID NO: 12), with capital letters representing exons of the synthetic gene and lowercase letters representing intron sequences of the synthetic gene; and also provided is the heavy chain amino acid sequence of the anti-CD40L antibody JB5-K74R (SEQ ID NO: 13).

[0030] Fig.11A synthetic nucleic acid sequence encoding the heavy chain of the anti-CD40L antibody JB5-K74R is provided (SEQ ID NO: 14), with capital letters representing exons of the synthetic gene and lowercase letters representing intronic sequences of the synthetic gene.

[0031] Fig.12 Provided are the amino acid sequences of the CDRs of the heavy and light chains of the anti-CD40L antibody JB5 (SEQ ID NOs: 15-20, respectively) and the amino acid sequence of the heavy chain of hu5C8 (SEQ ID NO: 21).

[0032] Fig.13A is a bar graph showing the ranked potency of 16 antibody clones compared to the anti-CD40L antibody 5c8. The ranked potency is IC50 clone / IC50 x 100 of 5c8.

[0033] Fig. 13B is a bar graph showing the ranked potency of 16 antibody clones compared to the anti-CD40L antibody AT- 1501. Rank potency is IC50 clone / IC50 x 100 of AT-1501.

[0034] Figure 14A-14G and Figure 14I-Figure 14Q The binding curves of antibodies from each clone are shown along with the binding curve of 5c8. Fig.14H Binding curves of AT-1501 and 5c8 are shown.

[0035] Figure 15A-15G and Figure 15I-Figure 15Q The binding curves of antibodies from each clone are shown along with the binding curve of AT-1501. Fig.15H Binding curves of AT-1501 and 5c8 are shown.

[0036] Fig.16A and Fig. 16B ] are graphs each showing binding of eight antibody clones, 5c8, AT-1501 and Abatacept to FcγRI. The only antibody with significant binding was the 5c8 antibody.

[0037] Fig.17A and Fig. 17B ] are graphs each showing binding of eight antibody clones, 5c8, AT-1501 and Abatacept to FcγRIIa. The only antibody with significant binding was the 5c8 antibody.

[0038] Fig.18A and Fig.18B] are graphs each showing binding of eight antibody clones, 5c8, AT-1501 and Abatacept to FcγRIIIa. No antibody showed significant binding to FcγRIIIa.

[0039] Fig. 18C and Fig.18D ] are graphs each showing binding of eight antibody clones, 5c8, AT-1501 and abatacept to FcγRIIIb. No antibody showed significant binding to FcγRIIIb.

[0040] Fig.19A and Fig.19B are graphs, each showing the binding of eight antibody clones, 5c8, AT-1501 and Abatacept to C1q. The only antibody with significant binding to C1q was the 5c8 antibody.

[0041] Figures 20A-20D is a graph of changes in IgAN-related proinflammatory markers in human subjects 12 weeks after renal transplantation. "*" indicates data with p values ​​between 0.05 and 0.01; "**" indicates data with p values ​​between 0.01 and 0.001; "***" indicates data with p values ​​less than 0.001. The data plotted are for IgAN-related proinflammatory markers IgA ( Fig. 20A )、IgE( Fig. 20B )、CD40L( Fig. 20C ) and C3( Fig.20D ).

[0042] Figure 21A-Figure 21D is a graph of changes in general transplantation pro-inflammatory markers in human subjects 12 weeks after renal transplantation. "*" indicates data with a p-value between 0.05 and 0.01; "**" indicates data with a p-value between 0.01 and 0.001; "***" indicates data with a p-value less than 0.001. The data plotted are for the general transplantation-related pro-inflammatory marker CXCL9 ( Fig.21A )、CXCL10( Fig. 21B )、IgM( Fig. 21C ) and C3( Fig.21D ).

[0043] Fig. 22 A non-limiting example schematic diagram depicts the study design and data output for Phase 1b and Phase 2 clinical trials, as discussed in Example 2.

[0044] Fig.23 Depicted are the demographics of the first four participants used in the Phase 1b clinical trial, as discussed in Example 2.

[0045] Fig.24Depicted is a non-limiting example of changes in renal function over time in patients following kidney transplantation and treatment, as monitored by eGFR levels, as discussed in Example 2.

[0046] Fig.25 Depicted are non-limiting example distributions for a cox proportional hazards model of eGFR levels and graft loss in human participants, as discussed in Example 2. DETAILED DESCRIPTION

[0047] Reference is made to the accompanying drawings forming a part thereof in the following detailed description. Unless the context otherwise dictates, in the accompanying drawings, similar symbols typically identify similar components. The illustrative embodiments described in the detailed description, drawings, and claims are not intended to be restrictive. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that, as generally described herein and shown in the accompanying drawings, aspects of the present disclosure may be arranged, replaced, combined, separated, and designed in a variety of different configurations, all of which are expressly contemplated herein. All references cited herein are expressly incorporated herein by reference in their entirety, and are used for the specific disclosures cited herein.

[0048] The present disclosure describes methods of treating patients with autoimmune disorders, such as autoimmune disorders associated with the kidney, including, for example, kidney disease, disorders or complications, by administering a therapeutically effective amount of a compound that blocks the interaction of CD40 with CD40 ligand (CD40L). The present disclosure also describes methods of treating patients by co-administering a compound that blocks the interaction of CD40 with CD40L with a compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art. Unless otherwise indicated, all patents, applications, published applications and other publications cited herein are incorporated herein by reference in their entirety. Unless otherwise indicated, in the case where there are multiple definitions for a term herein, those in this section shall prevail.

[0050] Those of ordinary skill in the art will appreciate that starting materials, biological and chemical materials, biological and chemical reagents, synthetic methods, purification methods, analytical methods, assay methods, and biological methods other than those specifically exemplified can be used to practice the present disclosure without resorting to undue experimentation. All functional equivalents known in the art for any such materials and methods are intended to be included in the present disclosure.

[0051] As used herein, "a" or "an" may mean one or more than one or more than one.

[0052] As used herein, the terms "about" or "approximately" have their ordinary meaning as understood by those skilled in the art, thereby indicating that a value includes the inherent variation of error in the method being employed to determine the value or the variation that exists among multiple determinations.

[0053] The use of the conjunction "or" is used interchangeably with "at least one of which". For example: in the case where a composition comprises A or B, the composition must comprise at least one of A and B, but may also comprise both A and B. Similarly, a composition comprising "A, B, C or D" must comprise at least one of the group of A, B, C and D, but may also comprise all or any combination of A, B, C and D.

[0054] Amino acid substitutions are indicated by convention wherein the original amino acid, the position of the amino acid in the designated sequence, and the replacement amino acid are identified, for example C11S would indicate that the cysteine ​​at position 11 of the polypeptide sequence is substituted with serine.

[0055] Humanized antibodies are antibodies produced from non-human species whose protein sequences have been modified to increase their similarity to naturally occurring antibody variants in humans. The "humanization" process is often applied to the development of monoclonal antibodies for administration to humans (e.g., antibodies developed as anti-cancer drugs).

[0056] At present, non-human antibodies are usually humanized by inserting the relevant CDR from the antibody produced in non-human animals into the human antibody "scaffold". The "direct" generation of humanized antibodies can be completed by inserting the appropriate CDR coding segment (responsible for the required binding properties) into the human antibody "scaffold". This can be achieved by using the recombinant DNA method of the appropriate carrier and expressing in mammalian cells. That is to say, after the antibody is developed to have the required properties in mice (or other non-humans), the DNA encoding the antibody can be separated, cloned into a vector and sequenced. The DNA sequence corresponding to the antibody CDR can then be determined. Once the precise sequence of the required CDR is known, it is possible to design a strategy for appropriately inserting these sequences into the construct containing the DNA of the human antibody variant. CDR can also be changed before being inserted into the scaffold, for example to increase specificity.

[0057] The term human antibody refers to an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a person and / or having been prepared using any technology for preparing human antibodies. This definition of human antibodies excludes humanized antibodies comprising non-human antigen binding residues in particular. As used herein, the term "variant" is defined as a modified or altered form of a wild-type sequence, such as one or more amino acids which may be replaced by other one or more amino acids or one or more non-amino acids that do not substantially significantly affect function. In certain embodiments, variants may contain an altered side chain of at least one amino acid residue.

[0058] The term "antigen" as used herein is defined as an entity that can stimulate the production of antibodies and specifically bind to them and / or an entity that triggers an immune system response. For example, a cell surface protein or a specific linear or nonlinear portion thereof. The term herein can be abbreviated as "Ag".

[0059] "Antigen-binding antibody fragment" refers to a molecule that is not an intact antibody, which comprises a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fd fragments, dAb fragments, Fab'-SH, F(ab')2; diabodies; triabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments, minimal recognition units composed of amino acid residues that mimic the hypervariable region of antibodies (e.g., isolated complementary determining regions (CDRs), such as CDR3 peptides). Antigen-binding fragments as disclosed in the present application bind to the antigen CD40L.

[0060] The antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain can be of any size or any amino acid composition, and will generally comprise at least one CDR, which is adjacent to or in frame with one or more framework sequences. In an antigen-binding fragment having a VH domain associated with a VL domain, the VH and VL domains can be in any suitable arrangement relative to each other. For example, the variable region can be dimeric and contain VH-VH, VH-VL or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody can contain a monomeric VH or VL domain.

[0061] Isolated antibodies include bispecific antibodies, in which each arm of the antibody or antigen-binding fragment binds to a different target or epitope.

[0062] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to a native antibody structure or having heavy chains that contain an Fc region.

[0063] "5c8" refers to a mouse anti-human antibody that binds CD40L and is produced by a hybridoma that is available from the ATCC under accession number HB 10916 and described in U.S. Pat. No. 5,474,771. "hu5c8" refers to a humanized form of 5c8, the sequence of which is disclosed in Karpusas et al., Structure, Vol. 9, pp. 321-329, (2001).

[0064] The term "specific binding" and the like means that the antibody or its antigen-binding fragment forms a relatively stable complex with the antigen under physiological conditions. Specific binding can be characterized by an equilibrium dissociation constant (KD) of about 3000 nM or less (i.e., a smaller KD indicates tighter binding); about 2000 nM or less; about 1000 nM or less; about 500 nM or less; about 300 nM or less; about 200 nM or less; about 100 nM or less; about 50 nM or less; about 1 nM or less; or about 0.5 nM. Specific binding to a particular antigen or epitope can be exhibited, for example, by an antibody having a KD for the antigen or epitope of at least about 1 x 10 -4 M, at least about 1x 10 -5 M, at least about 1x10 -6 M, at least about 1x 10 -7 M, at least about 1x 10 -8 M, at least about 1x 10 -9 M, alternatively at least about 1 x 10 -10 M, at least about 1x 10 -11 M, at least about 1x 10 -12 5,000, 10,000, or more, wherein KD refers to the equilibrium dissociation constant of a specific antibody-antigen interaction. Typically, an antibody that specifically binds to an antigen will have a KD that is 20 times, 50 times, 100 times, 500 times, 1000 times, 5,000 times, 10,000 times, or more, greater than an antigen or epitope. In addition, specific binding to a specific antigen or epitope can be exhibited, for example, by an antibody having a Ka to an antigen or epitope that is at least 20 times, 50 times, 100 times, 500 times, 1000 times, 5,000 times, 10,000 times, or more, greater than an epitope relative to a control, wherein Ka refers to the association rate of a specific antibody-antigen interaction.

[0065] The term "neutralizing antibody" includes antibodies that are capable of inhibiting and / or neutralizing the biological activity of CD40L, such as anti-CD40L antibodies or antigenic fragments thereof that inhibit or prevent or attenuate the binding of CD40L to CD40 and thereby inhibit or reduce the signaling pathway triggered by CD40L and / or inhibit or reduce the binding of CD40L to CD40.

[0066] The terms "antagonist antibody" or "antagonist antibody" are used equivalently herein and include antibodies that are capable of inhibiting and / or neutralizing the biological signaling activity of CD40L, as described above for neutralizing antibodies.

[0067] The terms and expressions that have been adopted herein are used as illustrative and not limiting terms, and it is not intended that any equivalents of the features shown and described or a portion thereof be excluded by the use of such terms and expressions, but it will be recognized that various modifications are possible within the scope of the methods and compositions disclosed herein. Therefore, it should be understood that although aspects of the present disclosure have been specifically disclosed by various embodiments that may include preferred embodiments, exemplary embodiments, and optional features, those skilled in the art may resort to modifications and variations of the concepts disclosed herein. Such modifications and variations are considered to be within the scope of the embodiments of the present disclosure as described and as may be defined by the appended claims.

[0068] In this specification, the identity percentage between polypeptide or amino acid sequence is mentioned.The identity percentage between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps and the length of each gap, and it is necessary to introduce them for the optimal comparison of two sequences.Identity can be measured as "local identity" or "overall identity".Local identity refers to the degree of sequence correlation between polypeptides, as determined by the matching between such sequence strings.Overall identity refers to the degree of sequence correlation of polypeptides compared with the full length of the reference polypeptide.Unless otherwise indicated, as used herein, identity means overall identity.For the purpose of this disclosure, the percentage of overall identity is calculated using the Blossum 62 scoring matrix (gap penalty is 12, gap extension penalty is 4, and frameshift gap penalty is 5) using Needleman and Wunsch ((1970) J.Mol.Biol. [Molecular Biology] 48:444-453) algorithm. There are a number of publicly available software programs that incorporate the Needleman and Wunsch algorithm, such as the GAP program in the GCG software package.

[0069] CD40L is also known as CD154, gp39, T-BAM, 5c8 antigen, or TNF-related activator protein (TRAP).

[0070] Throughout this specification, unless the context requires otherwise, the words "comprise", "comprises" and "comprising" will be understood to mean including a stated step or element or group of steps or elements, but not excluding any other step or element or group of steps or elements. "Consisting of" means including and limited to whatever follows the phrase "consisting of". Thus, the phrase "consisting of" indicates that the listed elements are required or mandatory, and that other elements may not be present. "Consisting essentially of" means including any element listed after the phrase, and is limited to other elements that do not interfere with or contribute to the activity or action specified in the present disclosure of the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are required or mandatory, but other elements are optional and may or may not be present, depending on whether they substantially affect the activity or action of the listed elements.

[0071] The terms "function" and "functionality" as used herein have their plain and ordinary meaning as understood in light of the present specification, and refer to a biological function, an enzyme function, or a therapeutic function.

[0072] As used herein, the term "yield" of any given substance, compound, or material has its simple and ordinary meaning as understood in accordance with this specification, and refers to the actual total amount of the substance, compound, or material relative to the expected total amount. For example, the yield of a substance, compound, or material is, is about, is at least, is at least about, is not more than, or is not more than about 80%, 81%, 82%, 83%, 84%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, including all decimals therebetween, of the expected total amount. The yield may be affected by the efficiency of the reaction or process, unwanted side reactions, degradation, the amount of input substance, compound, or material, or the loss of the desired substance, compound, or material during any step of the production.

[0073] As used herein, the term "isolated" has its plain and ordinary meaning as understood in light of this specification, and refers to substances and / or entities that have been (1) separated from at least some of the components with which they were originally associated (whether in nature and / or in an experimental setting), and / or (2) produced, prepared and / or manufactured by the hand of man. Isolated substances and / or entities can be separated from other components with which they were originally associated by equal to, about, at least, at least about, not greater than, or not greater than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98%, about 99%, substantially 100%, or 100% (or a range including and / or spanning the foregoing values). In some embodiments, the isolated agent is, is about, is at least, is at least about, is not more than, or is not more than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, substantially 100%, or 100% (or a range including and / or spanning the above values) pure. As used herein, an "isolated" substance can be "pure" (e.g., substantially free of other components). As used herein, the term "isolated cell" can refer to a cell that is not contained in a multicellular organism or tissue.

[0074] The term "isolated protein" or "isolated polypeptide" (e.g., an isolated antibody or isolated antigen-binding fragment) is a protein or polypeptide that, by its origin or source of derivation, is not associated with naturally associated components that accompany it in its native state; is substantially free of other proteins from the same species; is expressed by cells from a different species; or does not occur in nature. Thus, a polypeptide that is chemically synthesized or synthesized in a cell system different from the cell from which it is naturally derived will be "isolated" from its naturally associated components. A protein can be rendered substantially free of naturally associated components by separation using protein purification techniques well known in the art.

[0075] As used herein, "in vivo" is given its plain and ordinary meaning as understood in light of this specification, and refers to methods performed within living organisms (typically animals, mammals (including humans) and plants) or living cells that make up these living organisms (as opposed to tissue extracts or dead organisms).

[0076] As used herein, "ex vivo" is given its plain and ordinary meaning as understood in light of this specification, and refers to a method performed outside a living organism with little change to natural conditions.

[0077] As used herein, "in vitro" is given its plain and ordinary meaning as understood in light of this specification, and refers to methods performed outside of biological conditions (eg, in a culture dish or test tube).

[0078] As used herein, "nucleic acid", "nucleic acid molecule" or "nucleotide" refers to a polynucleotide or oligonucleotide, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), an oligonucleotide, a fragment produced by the polymerase chain reaction (PCR), and a fragment produced by any of ligation, cleavage, endonuclease action, exonuclease action, and synthesis. Nucleic acid molecules can be composed of monomers that are naturally occurring nucleotides (such as DNA and RNA) or analogs of naturally occurring nucleotides (e.g., enantiomeric forms of naturally occurring nucleotides) or a combination of both. Modified nucleotides can have changes in the sugar moiety and / or the pyrimidine or purine base moiety. Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyls, amines, and azido groups, or sugars can be functionalized as ethers or esters. In addition, the entire sugar moiety can be replaced by a spatially and electronically similar structure (such as azasugars and carbocyclic sugar analogs). Examples of modifications in the base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes. Nucleic acid monomers can be connected by phosphodiester bonds or analogs of such connections. Phosphodiester-connected analogs include phosphorothioates, phosphorodithioates, selenophosphates, diselenphosphates, aniline thiophosphates, phosphoranilidates, phosphoramidates, etc. The term "nucleic acid molecule" also includes so-called "peptide nucleic acids", which include naturally occurring or modified nucleic acid bases attached to a polyamide backbone. Nucleic acids can be single-stranded or double-stranded.

[0079] The terms "peptide", "polypeptide" and "protein" as used herein have their simple and ordinary meanings as understood in light of this specification, and refer to macromolecules composed of amino acids linked by peptide bonds. Many functions of peptides, polypeptides and proteins are known in the art, and include, but are not limited to, enzymes, structures, transport, defense, hormones or signal transduction. Peptides, polypeptides and proteins are usually, but not always, biologically produced by ribosome complexes using nucleic acid templates, although chemical synthesis is also available. By manipulating nucleic acid templates, peptide, polypeptide and protein mutations (such as substitutions, deletions, truncations, additions, duplications or fusions of more than one peptide, polypeptide or protein) can be performed. These fusions of more than one peptide, polypeptide or protein can be contiguously linked in the same molecule, or with additional amino acids (e.g., linkers, repeat sequences, epitopes or tags) therebetween, or any other sequence that is, is about, is at least, is at least about, is no more than, or is no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 bases long, or any length within the range defined by any two of the above lengths. The term "downstream" as used herein with respect to a polypeptide has its simple and common meaning as understood in accordance with this specification, and refers to the sequence after the C-terminus of the preceding sequence. The term "upstream" as used herein with respect to a polypeptide has its plain and ordinary meaning as understood in light of the present specification, and refers to a sequence preceding the N-terminus of a subsequent sequence.

[0080] The term "gene" as used herein has its simple and common meaning as understood according to this specification, and generally refers to a part of a nucleic acid encoding a protein or functional RNA; however, the term may optionally encompass regulatory sequences. It will be understood by those of ordinary skill in the art that the term "gene" may include gene regulatory sequences (e.g., promoters, enhancers, etc.) and / or intron sequences. It will be further understood that the definition of a gene includes reference to nucleic acids that do not encode proteins but encode functional RNA molecules (such as tRNA and miRNA). In some cases, a gene includes regulatory sequences that are involved in transcription or information production or composition. In other embodiments, a gene comprises a transcribed sequence encoding a protein, polypeptide, or peptide. Consistent with the terms described herein, an "isolated gene" may include one or more transcribed nucleic acids, regulatory sequences, coding sequences, etc. that are substantially separated from other such sequences (such as other naturally occurring genes, regulatory sequences, polypeptide or peptide coding sequences, etc.). In this regard, the term "gene" is used to simply refer to a nucleic acid comprising a transcribed nucleotide sequence and its complementary sequence. As will be understood by those skilled in the art, this functional term "gene" includes genomic sequences, RNA or cDNA sequences, or smaller engineered nucleic acid segments, including nucleic acid segments of non-transcribed portions of genes, including but not limited to non-transcribed promoter or enhancer regions of genes. Smaller engineered gene nucleic acid segments can express or can be adapted to express proteins, polypeptides, domains, peptides, fusion proteins, mutants and / or the like using nucleic acid manipulation techniques.

[0081] As used herein, the terms "individual", "subject", "host" or "patient" have their usual meaning as understood by those skilled in the art, and thus include human or non-human mammals. The term "mammal" is used in its usual biological sense. Thus, it specifically includes, but is not limited to, primates (including simians (chimpanzees, apes, monkeys), humans), cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rodents, rats, mice or guinea pigs.

[0082] The dimensions and values ​​disclosed herein should not be understood as being strictly limited to the exact numerical values ​​recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as "20 mm" is intended to mean "about 20 mm".

[0083] The following abbreviations are used herein: superoxide dismutase-1 (SOD1); T cell receptor (TCR); major histocompatibility complex (MHC); antigen presenting cell (APC); phosphate buffered saline (PBS); complementarity determining region (CDR). "IP" means intraperitoneal, and "IV" means intravenous.

[0084] R1 is a hamster monoclonal antibody that binds to the mouse CD40 ligand. "Wild type" as used herein means a non-transgenic mouse. "Small molecule" as used herein means a compound with a molecular weight of less than 2000 Daltons. "Treatment" or "treating" as used herein includes preventive treatment and therapeutic treatment. The phrase "therapeutically effective amount" means an amount of a compound or a pharmaceutically acceptable salt thereof sufficient to inhibit or allow improvement in a disorder or condition to be treated by administration alone or in combination with another agent or treatment in a specific subject or subject population.

[0085] An "effective amount" of an agent (eg, a pharmaceutical formulation) refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

[0086] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the equilibrium dissociation constant (KD) (k off / k on KD and affinity are inversely related. The KD value is related to the concentration of the antibody (the amount of antibody required for a particular experiment), so the lower the KD value (the lower the concentration), the higher the affinity of the antibody. Affinity can be measured by conventional methods known in the art (including those described herein). Specific, illustrative and exemplary embodiments for measuring binding affinity can be by radioimmunoassay (RIA); Surface plasmon resonance (SPR) on a GE Healthcare Europe GmbH, Zurich, Switzerland instrument (GE Healthcare Europe GmbH, Zurich, Switzerland) was measured using human CD40 ligand polypeptide as the analyte by capturing the antibody on a protein A-coupled CM5 research grade sensor chip (GE Healthcare Europe GmbH, Zurich, Switzerland; BR-1000-14). Other methods may include radioimmunoassay and kinetic exclusion assay. Kinetic exclusion assay is a general immunoassay platform that is capable of measuring the equilibrium dissociation constant as well as the association rate constant and dissociation rate constant of antigen / antibody interactions.

[0087] TMB is the abbreviation of 3,3′,5,5′-tetramethylbenzidine.

[0088] "CDR domain" as used herein means the complementarity determining region of an antibody with or without flanking sequences.

[0089] "Effector function" refers to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement dependent 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. An effector function is the ability of the Fc or constant region of an antibody to bind to proteins including but not limited to the following: Fc receptors (FcR) (e.g., high affinity IgG Fc region receptor Ia (FCγRIa) (CD64), low affinity immunoglobulin γ Fc region receptor IIa (FCγRIIa) (CD32), low affinity immunoglobulin γ Fc region receptor IIIa (FCγRIIIA) (CD16a), low affinity immunoglobulin γ Fc region receptor IIIb (FCγRIIIb) (CD16b)). In some embodiments, where the antibody and antigen-binding fragment thereof have an Fc domain, the Fc domain has been engineered to reduce or eliminate one or more Fc effector functions. In a preferred embodiment, the Fc domain has been engineered to reduce or eliminate platelet activation and / or platelet aggregation and the attendant risk of thromboembolism.

[0090] Identification of candidate compounds

[0091] The co-stimulatory pathway, along with other interactions, involves the binding of CD40 on B cells to CD40L (also known as CD154, gp39, T-BAM, 5c8 antigen, CD40CR and TRAP) on T cells. Human CD40 is expressed on mature B cells as well as macrophages, dendritic cells, fibroblasts and activated endothelial cells. Blockade of CD40:CD40L binding is thought to promote the formation of type I T helper cell responses.

[0092] Using compounds that block these interactions and inhibit costimulatory signals, a large amount of work has demonstrated the immunomodulatory effects of blocking one or more of CD40L, CD80 or CD86 in preclinical models of transplantation and autoimmunity. Blocking CD40L function with blocking antibodies or expressing CD40L-Ig with adenovirus increases the allograft transplantation period by 30 to 90 days. Similar studies that block CD80 / CD86 on APCs with CTLA4-Ig or express CTLA4-Ig with adenovirus transiently increase allograft transplant survival. Transplant rejection in these models is transient, and graft rejection ensues over time. Long-term suppression of transplant rejection can be achieved by blocking costimulatory pathways with CTLA4-Ig and blocking CD40L activation of APCs with anti-CD40L antibodies.

[0093] Blocking antibodies against CD40L or genetic deletion of CD40L in mice have demonstrated that CD40L improves disease progression, survival, and surrogate markers of disease in preclinical models of experimental allergic encephalomyelitis (EAE), a model of multiple sclerosis, collagen-induced arthritis, and systemic lupus. Blockade of CD40:CD40L binding appears to reduce the ability of macrophages to produce nitric oxide, which mediates many of the pro-inflammatory activities of macrophages.

[0094] From such studies it appears that blocking the CD40:CD40L interaction and / or blocking the CD28:CD80 or CD28:CD86 interaction can modulate the immune response.

[0095] The immunohistochemical data showed good correlation with the gene expression data and identified macrophages in G93A mice as antigen-presenting cells that infiltrate skeletal muscle during disease progression.

[0096] In order to determine the presence and location of potential antigen presenting cells in the gastrocnemius from G93A and wild-type animals, immunohistochemistry was performed on the gastrocnemius tissue harvested from G93A and wild-type mice on the 110th day. The tissue was embedded in OCT immediately after harvesting. The frozen sections were stained with H&E and hybridized with antibodies against myelin (anti-S100b antibody) or antibodies against hematopoietic cell lineages (including T cells (anti-CD3 antibody), B cells (CD45R pan-B cell antibody) and macrophages (anti-CD11b antibody). At the 110th day, there was CD11b positive macrophage infiltration, and the macrophages appeared to be located in the axons of the nerves that innervate skeletal muscle. The location of macrophages is not scattered throughout the muscle, which indicates that inflammation is not due to muscle atrophy or muscle fiber remodeling.

[0097] In order to confirm the identity of mononuclear cell lineage cells in skeletal muscle at the 110th day and to illustrate the relevance of these cells compared with non-transgenic animals, additional immunohistochemistry was performed with a group of antibodies specific to the macrophage lineage. Anti-S100b antibody was used to mark the myelin-related axons that dominate skeletal muscle. All macrophage-specific antibodies (anti-CD11b antibody, anti-CD86 antibody and anti-MAC1 antibody) localized macrophages to the axons of the nerves that dominate the skeletal muscle of G93A mice, while there were no macrophages on the axons of wild-type animals. The location of macrophages is unique to the nerves that dominate muscle, and there are no macrophages on the muscle fibers of muscle.

[0098] Gene expression data show that the genes related to the costimulatory pathway in both spinal cord and skeletal muscle are temporarily increased during disease progression. In order to characterize the time of macrophage infiltration into skeletal muscle, immunohistochemistry was performed to the gastrocnemius sections from the 60th day, the 80th day and the 100th day in G93A mice. There was no evidence of macrophage infiltration and positioning in axons at the 60th day. Macrophages were obvious at the 80th day, and as previously described for skeletal muscle at the 110th day, they were positioned in the axons of dominance muscle. The number of macrophages increased between the 80th day and the 100th day, and the accumulation of macrophages was unique to the axons of dominance skeletal muscle.

[0099] To quantify the increase in macrophage infiltration, representative sections from 5 G93A and 5 wild-type animals were hybridized with anti-CD86 antibodies and the number of macrophages per 10,000 square microns was counted. The number of macrophages was the same in wild-type animals at the 60, 80, and 110 day time points. Macrophages transiently accumulated in skeletal muscle between days 80 and 100, and very few macrophages were present in wild-type skeletal muscle.

[0100] In summary, the immunohistochemical data correlated very well with the gene expression data and identified macrophages as antigen presenting cells that infiltrate skeletal muscle during disease progression in the G93A mouse model. An unexpected finding was that the macrophage infiltration appeared to specifically target axons innervating skeletal muscle, as localized by labeling with antibodies against myelin and macrophages.

[0101] A matrix-matched non-competitive enzyme-linked immunosorbent assay (ELISA) in a sandwich format was used to determine tissue levels of MR1 against murine CD40L. A seven-point standard curve was included on each plate. Standards were prepared using purified MR1 spiked into a PBS diluent solution. The PBS diluent solution was matrix-matched to normal mouse tissue at equal dilutions of unknown samples to correct for any nonspecific effects caused by tissue lysates.

[0102] In both female and male G93A mice, 84 plasma samples were collected over a two-week period following dosing (10 mg / kg, IP) for pharmacokinetic analysis.

[0103] The elimination half-life was similar in females (23 days) and males (22 days) and is similar to the half-life of a typical mouse IgG2-based antibody in mice. No evidence of anti-hamster antibody responses was observed.

[0104] Females showed a slightly smaller volume of distribution for MR1 compared to males, and therefore showed higher plasma levels when given the same 10 mg / kg dose. Males showed faster clearance and a higher volume of distribution. Therefore, males would require a higher dose to achieve similar plasma levels. Figure 1A MR1 concentration over time is shown using a linear concentration (Y) axis. Figure 1B MR1 concentration over time is shown using the log concentration (Y) axis.

[0105] Thirty-six female G93A mice were litter-matched and randomly divided into two study groups. 18 G93A mice were placed in the MRI treatment group, and the other 18 G93A mice were placed in the control group. The study days were based on days from birth.

[0106] A single injection of 56 ug MR1 was administered intraperitoneally (IP) on day 50. After the bolus, weekly maintenance injections of 18 ug MR1 were administered by intraperitoneal injection. Doses were prepared in vehicle (phosphate buffered saline (PBS, pH 7.3)) to a total volume of 200 μl. Control animals were administered 200 μl PBS. Starting on day 54, animals were monitored daily throughout the study and daily body weight measurements were performed.

[0107] Mutant SOD1 transgenic animals exhibit normal body weight (BW) characteristics as neonates and gain weight normally into adulthood compared to non-transgenic animals. Depending on the nature of the genetic mutation in the transgene and the number of copies of the mutant transgene, weight loss becomes evident in adult animals and persists until death. Analysis of weight loss in treatment and control groups can provide insights into the effects of putative treatments on the rate of disease onset and progression. To assess the effects of MR1 treatment on body weight, two summary parameters were examined: (1) the change in BW from the start of the study to peak body weight, which can reflect the effect on disease onset; and (2) the change in BW from peak body weight to death, which can reflect the effect on disease progression.

[0108] The time-to-event curves from day 40 to the time to peak body weight for the comparative MR1-treated and control groups are shown in Figure 2 The median time to peak body weight was 50 days in the control group compared with 51 days in the MR1-treated group. This difference was not significant when analyzed by Kaplan-Meier using log-rank and Wilcoxon statistical models, Cox proportional hazards models, or parametric statistical tests.

[0109] The time-to-event curves from peak body weight to death for the comparative MR1-treated and control groups are shown in Figure 3In animals treated with MRI, the time from peak body weight to death was statistically significantly later by 15 days. The risk of control animals dying sooner after reaching peak body weight was 2.4 to 4.7 times higher than that of animals treated with MR1. The median time from peak body weight to death in the control group was 26 days, while it was 41 days in the MR1-treated group. The significance of each analysis in this example was calculated in several ways to better evaluate the significance. The delay was statistically significant when analyzed using several methods. (Kaplan-Meier, log-rank, p=0.0110 and Wilcoxon, p=0.0069; Cox proportional hazards model, p=0.05151; parametric statistical model, p=0.0122). Based on the body weight data, MR1 appears to have a small effect on the onset of disease in the G93A mouse model, but has a significant effect in slowing the rate of weight loss from peak body weight until death.

[0110] The survival time of treated animals was 13 days later than that of control animals. The time-event curves from peak body weight to death for the comparative MR1 treated and control groups are shown in Figure 4 The median survival time for the control group was 128 days, and the median survival time for the MR1-treated group was 141 days. The risk of faster death in control animals was 2.8 to 3.2 times higher than that of animals treated with MR1. The delay was statistically significant when analyzed using several methods. (Kaplan-Meier, log-rank (p=0.0040) and Wilcoxon tests (p=0.0109); Cox proportional hazards model (p=0.0060); parametric analysis (p=0.0049)). 60 female and 36 male G93A mice were litter-matched and randomly assigned to treatment or control groups. 30 female mice and 18 male mice were treated with MR1 starting on day 50. Study days are based on birth.

[0111] On day 50, a single bolus of 5.22 mg / kg or 6.75 mg / kg MR1 was administered intraperitoneally in females or males, respectively. After the bolus, females received weekly injections of 1 mg / kg MR1 and males received weekly injections of 1.34 mg / kg MR1 via intraperitoneal injection. Doses were prepared in vehicle (phosphate buffered saline (PBS, pH 7.3)) to a total volume of 200 μl. Control animals were administered 200 μl PBS. Animal weight and euthanasia criteria were monitored as previously described.

[0112] The time-to-event curves from day 40 to the time to peak body weight for the comparative MR1-treated and control groups are shown in Figure 5AThe median time to peak body weight was 49 days in the control group compared with 53 days in the MR1-treated group. This difference was not significant when analyzed by Kaplan-Meier using log-rank and Wilcoxon statistical models, Cox proportional hazards models, or parametric statistical tests.

[0113] The time-to-event curves from peak body weight to death for the comparative MR1-treated and control groups are shown in Figure 5B In the MRI-treated animals, the time from peak body weight to death was statistically significantly delayed by 6 days. The median time from peak body weight to death in the control group was 29 days, while it was 35 days in the MR1-treated group. The significance of each analysis in this example was calculated in several ways to better assess the significance. The delay was statistically significant when analyzed using several statistical models: Kaplan-Meier, log-rank (p=0.0413) and Wilcoxon (p=0.0732); and Cox proportional hazards model (p=0.0460). Based on the body weight data, MR1 appears to have a small effect on the onset of disease in the G93A mouse model, but has a significant effect in slowing the rate of weight loss from peak body weight until death.

[0114] The time-to-event graphs of the age at which MR1 and control animals progressed to a neurologic score of 2 and the number of days to a neurologic score of 2 are shown in Figure 5C The time to score 2 was 113 days in the control group and 121 days for the MR1 treated group. Based on the neurological function score data, MR1 delayed the onset of disease in the G93A mouse model by approximately 8 days, and the delay was statistically significant when analyzed using the following analytical models: Kaplan-Meier, log-rank (p=0.0038) and Wilcoxon (p=0.0017); and Cox proportional hazards model (p=0.0010).

[0115] The survival of treated animals was 9 days later than that of control animals. The median survival of the control group was 124 days, and the median survival of the MR1 treated group was 133 days. Figure 5D The delay was statistically significant when analyzed using several statistical models: Kaplan-Meier, log-rank (p=0.0043), and Wilcoxon tests (p=0.0040); and the Cox proportional hazards model (p=0.0030).

[0116] It should be understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to those skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims.

[0117] The nucleic acid sequences encoding the heavy and light chains of the desired antibodies are designed to be suitable for expression in mammalian cells, such as Chinese hamster ovary (CHO) cells. The nucleic acids are then artificially synthesized and connected to the antibody expression vector BPJPuro using standard molecular biology techniques. BPJPuro is a dual-gene mammalian expression vector optimized for selective and stable expression of immunoglobulins in Chinese hamster ovary (CHO) cells. The vector is then transfected into CHO cells, and stable transfectants are selected.

[0118] Similarly, nucleic acid encoding the heavy chain having the amino acid sequence of SEQ ID NO: 9 (SEQ ID NO: 10) and nucleic acid encoding the light chain having the amino acid sequence of SEQ ID NO: 7 (SEQ ID NO: 8) were synthesized and ligated into the antibody expression vector BPJPuro.

[0119] The obtained expression vector encoding heavy chain and light chain is transfected into CHO system (CHO SA, Cellectis SA, Paris, France) using liposome-mediated transfection. Stable transfectants are separated by puromycin selection and subcloned to provide clonal cell lines. Candidate cell lines are adapted to serum-free suspension culture, and are screened for IgG production and robust growth. A cell line is selected and named JB5, which is cultivated in a pilot scale bioreactor, and antibody JB5 is purified from conditioned medium by sequential concentration, protein A / G affinity chromatography and size exclusion chromatography.

[0120] The binding kinetics of the JB5 antibody relative to the parent antibody hu5c8 were determined using a three-part sandwich ELISA assay. All washes were performed using 3 washes of 250 μl PBS. 96-well polystyrene plates were coated with 100 μl / well JB5 or hu5c8 antibodies (2 μg / ml) for 16 hours at 4°C. The plates were washed and then blocked with 2% bovine serum albumin / PBS for 1 hour at room temperature. The plates were washed and recombinant human CD40L protein (Santa Cruz Biotechnology, Santa Cruz, California, USA) was added to the plates and titrated by 2-fold dilution starting at 2000 ng / ml. After binding and washing, 100 μl of biotinylated goat anti-human CD40L polyclonal antibody (200 ng / ml) and 100 μl of streptavidin-horseradish peroxidase conjugate (100 ng / ml) were used to detect the bound CD40L protein. Colorimetric detection was performed using the colorimetric reagent TMB (3,3′,5,5′-tetramethylbenzidine), and the absorbance at 450 nm was analyzed spectrophotometrically. The resulting binding curve ( Figure 6) shows that JB5 (circles) has highly similar CD40L binding relative to the parent antibody hu5c8 (squares). The control protein CTLA4-IgG1 (triangles) with the same Fc domain as JB5 did not show significant binding. The calculated EC50s for hu5c8 and JB5 were 114 and 137 nM, respectively. JB5-R28K and JB5-K74R showed similar binding to that of JB5.

[0121] To compare the CD40L binding of antibodies from all 16 clones compared to 5c8 or AT-1501 (also known as tegoprubart, a humanized monoclonal antibody that inhibits the CD40 ligand), a binding assay was run using 2 clones (5c8 and AT-1501 run on the same 96-well assay plate). A three-part sandwich ELISA assay was used to determine the binding level of the antibodies of the present disclosure compared to reference antibodies 5c8-19 and AT-1501. 96-well polystyrene plates were coated with recombinant human CD40L (BioLegend catalog number 591706) (using 2 μg / ml in PBS) and 50 μl / well was added to a Costar 96-well 1 / 2 area high binding assay plate (Corning 3690) and incubated overnight at 4°C. The plate was blocked with (1X) PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature to prevent background binding. The binding curve of 5C8 or AT1501 (serial 2-fold dilution from 2 μg / ml) (50 μl / well) was added and incubated for 1 hour at room temperature. The plate was washed and incubated for 1 hour at room temperature with 1:10,000 dilution of HRP-(Fab2) donkey anti-human IgG (Fc specific) (Jackson Immuno. 709-036-098) (50 μl / well). The plate was washed and TMB substrate (Surmodics BioFX TMBW-1000-01) (50 μl / well) was added. After 5 minutes at room temperature, the color development was stopped with 25 μl 2NH2SO4. The plate was read on a Molecular Devices SpectraMax M5 plate reader using the SoftMax Pro 6.2.2 program to determine the absorbance at 450 nm.

[0122] The relative CD40L binding potency was calculated as follows: [IC50 (clone) / IC50 (5c8 or AT-1501) x 100%]. The ranked potency of the 16 clones compared to 5c8 is shown in Fig.13A and the ranking efficacy compared to AT-1501 is shown in Fig. 13B The resulting binding curve is shown in Figure 14A-Figure 14Q(Comparison of 16 clones with 5c8, Fig.14H shows a comparison between AT-1501 and 5c8) and Figure 15A-Figure 15Q (Comparison of 16 clones with AT-1501, Fig.15H The IC50, LCL, UCL and relative potency of each clone are shown in Table 1 (comparison with 5c8) and Table 2 (comparison with AT-1501).

[0123] Table 1:

[0124]

[0125]

[0126]

[0127] Table 2:

[0128]

[0129]

[0130] 16 VH / VL antibody clones were constructed using IgG1 Fc (SEQ ID NO: 21) with two mutations P238S and N297G. These antibody clones were assayed for Fc effector function binding to human FcγRI, FcγRIIa and FcγRIIIa.

[0131] Anti-CD40L antibodies (including abatacept as a negative control) were diluted to 2 μg / ml in (1X) PBS and 50 μl / well were added to Costar 96-well 1 / 2 area high binding assay plates (Corning 3690) for overnight incubation at 4°C. The plates were blocked with (1X) PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature to prevent background binding. A binding curve of recombinant human FcγRI, IIa, IIIa and IIIb (serial 2-fold dilutions starting from 5 μg / ml) (50 μl / well) was added and incubated for 1 hour at room temperature. The plate was washed and incubated at room temperature for 1 hour with 2 μg / ml of mouse anti-human CD16 (anti-FcRIII); CD32 (anti-FcRIIa) or CD64 (anti-FcRI) (eBioSciences / Invitrogen 14-0168-82; 16-0329-81; 14-0649-82) (50 μl / well). The plate was washed and incubated at room temperature for 1 hour with 1:10,000 dilution of HRP-(Fab2) goat anti-mouse IgG (Fc specific) (Jackson ImmunoResearch 116-036-071) (50 μl / well). The plate was washed and TMB substrate (Surmodics BioFX TMBW-1000-01) (50 μl / well) was added. After 5 min at room temperature, the color development was stopped with (25 μl / well) 2NH2SO4. Plates were read on a Molecular Devices SpectraMax M5 plate reader using the SoftMax Pro 6.2.2 program to determine absorbance at 450 nm.

[0132] As can be observed in the figures, the clones disclosed in the present application were negative for binding to the three Fc receptors, whereas 5c8 bound to FcγRI, FcγRIIa, but not to FcγRIIIa or FcγRIIIb. Fig.16A and Fig. 16B Binding of eight antibody clones to FcγRIa is shown individually in comparison with the binding curves of 5c8, AT-1501 and abatacept. Fig.17A and Fig. 17B Binding of eight antibody clones to FcγRIIa is shown individually in comparison to the binding curves of 5c8, AT-1501 and abatacept. Fig.18A and Fig.18B Binding of eight antibody clones to FcγRIIIa is shown individually in comparison to the binding curves of 5c8, AT-1501 and abatacept. Fig. 18C and Fig.18DEach shows the binding of eight antibody clones to FcγRIIIb compared with the binding curves of 5c8, AT-1501 and abatacept. Fig.16A , Fig.17A , Fig.18A and Fig.18B Results from antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4 are shown; Fig. 16B , Fig. 17B , Fig. 18C and Fig.18D Results from antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2 are shown).

[0133] Anti-CD40L antibodies (including abatacept as a negative control) were diluted to 2 μg / ml in (1X) PBS and 50 μl / well were added to Costar 96-well 1 / 2 area high binding assay plates (Corning 3690) for overnight incubation at 4°C. The plates were blocked with (1X) PBS / 1.0% BSA (140 μl / well) for 1 hour at room temperature to prevent background binding. Native human C1q protein (Abcam ab96363) (serial 2-fold dilutions starting from 10 μg / ml) (50 μl / well) was added and the binding curve was continued for 1 hour at room temperature. The plates were washed and HRP-sheep anti-human C1q (Abcam ab46191) at a dilution of 1:400 was added (50 μl / well) and incubated for 1 hour at room temperature. The plate was washed and TMB substrate (Surmodics BioFX TMBW-1000-01) (50 μl / well) was added. After 5 minutes at room temperature, the color development was stopped with (25 μl / well) 2NH2SO4. The plate was read on a Molecular Devices SpectraMax M5 plate reader using the SoftMax Pro 6.2.2 program to determine the absorbance at 450 nm.

[0134] As can be observed in the figure, antibodies from all sixteen antibody clones disclosed in the present application were negative for binding to C1q, whereas 5c8 showed significant binding. Fig.19A and Fig.19B Binding of eight antibody clones to C1q is shown individually in comparison to the binding curves of 5c8, AT-1501 and abatacept. Fig.19A Results from antibody clones 4-4, 5-3, 6-6, 7-5, 8-3, 8-4, 10-1, and 10-4 are shown; Fig.19B Results from antibody clones 11-5, 12-4, 13-2, 15-1, 15-4, 16-3, 17-1, and 18-2 are shown.

[0135] Since genetic expression data indicated that costimulatory pathways were involved, the efficacy of MRI was evaluated in the G93A model. MR1 binds to CD40L, thereby blocking its interaction with CD40, which participates in costimulatory pathways involved in immune responses. It has been reported in the literature that MR1 is effective in treating rheumatoid arthritis and graft-versus-host disease, both of which have strong immune components. Rheumatoid arthritis is an autoimmune disease, and graft-versus-host disease occurs when the host's body mounts a strong immune response to the transplanted tissue.

[0136] Methods according to the present disclosure include methods of treating patients with autoimmune disorders (including kidney-related autoimmune disorders, such as kidney-related diseases, disorders or complications) by administering to the patient a compound that blocks the interaction of CD40L with CD40 and / or blocks the interaction of CD28 with CD80 and / or blocks the interaction of CD28 with CD86. In some embodiments, the methods include treating patients with colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, kidney transplantation, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis or IgA nephropathy by administering to the patient a compound that blocks the interaction of CD40L with CD40. In some embodiments, the methods include treating the patient by administering an anti-CD40L antibody. In some embodiments, the methods include treating a patient with an autoimmune disease. Non-limiting examples of autoimmune diseases include systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, and Graves' disease.

[0137] The therapeutic compounds provided herein include any compound that blocks the interaction of CD40 with CD40L. For example, a number of animal studies describe agents that can block CD40:CD40L binding (see, e.g., US2005158314 and US7173046, which are hereby incorporated by reference). Many anti-CD40L antibodies have been generated and characterized (see, e.g., U.S. Pat. No. 5,876,950 to Bristol-Myers Squibb, which is hereby incorporated by reference). Anti-CD40L antibodies that can be used in the methods of the present disclosure include, but are not limited to, MR1, a hamster monoclonal antibody available from Taconic (Hudson, NY) and BD Biosciences (San Jose, CA); 5c8, a humanized antibody described in U.S. Pat. No. 5,474,771 (which is hereby incorporated by reference); hamster-human chimeric antibody IDEC 131 / E6040, a humanized monoclonal antibody comprising a human gamma-1 heavy chain and a human kappa light chain having the CDRs of murine monoclonal antibody clone 24-31, commercially available from Ancell (Catalog No. X353-020, Bayport, MN); ABI 793; Sgn-40; ImxM90 (Imnis); ImxM91 (Imnis); ImxM92 (Imnis); and anti-CD40L mAb commercially available from Genzyme (Cambridge, Mass., catalog number 80-3703-01). Also commercially available is anti-CD40L mAb from PharMingen (San Diego, catalog number 33580D). Embodiments according to the present disclosure include methods of treating a patient suffering from a kidney-related autoimmune disease, disorder, or complication comprising administering a therapeutically effective amount of any compound capable of binding to CD40L, such as an anti-CD40L antibody. In some embodiments, the methods include treating a patient with a kidney-related autoimmune disease, disorder or complication, comprising administering a therapeutically effective amount of an anti-CD40L antibody selected from the group consisting of MR1, 5c8, IDEC 131 / E6040, clone 24-31, ABI 793, ImxM90, ImxM91, ImxM92 or Sgn-40. In some embodiments, the antibody is 5c8. In some embodiments, the antibody is MR1.

[0138] In some embodiments, the kidney-related disease, disorder or complication is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, kidney transplantation, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis or IgA nephropathy, by administering to the patient a compound that blocks the interaction of CD40L with CD40. In some embodiments, the method of treating a patient is by administering an anti-CD40L antibody. In some embodiments, the methods include treating a patient with an autoimmune disease. Non-limiting examples of autoimmune diseases include systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, and Graves' disease. In some embodiments, the anti-CD40L antibody is MR1. In some embodiments, the anti-CD40L antibody is 5c8.

[0139] In some embodiments, the method of treatment comprises administering a therapeutically effective amount of an anti-CD40 antibody. In some embodiments, the anti-CD40L compound is a Fab fragment, F(ab')2, F(ab'), a single-chain antibody, a polypeptide, a fusion construct of a polypeptide, etc. In some embodiments, these compounds are small molecule compounds capable of blocking CD40:CD40L interactions. In some embodiments, these compounds include BIO3417 or any compound disclosed in U.S. Patent No. 7,173,046 that has the ability to block CD40:CD40L interactions.

[0140] Compounds that block the CD40:CD40L interaction can be administered in combination with other compounds. Thus, in some embodiments, the methods of treating a patient as described herein include administering a combination of a compound that blocks the interaction of CD40L with CD40 and a compound that blocks the interaction between CD80 and CD28. In some embodiments, these methods include administering a combination of a compound that blocks the interaction of CD40L with CD40 and a compound that blocks the interaction between CD86 and CD28. In some embodiments, the compound that blocks the interaction of CD80 with CD28 is galiximab, or H1f1 and h3d1, or 16C10, or 7C10. In some embodiments, the compound that blocks the interaction between CD86 and CD28 is a CTLA4-Ig protein conjugate, such as abatacept or belatacept. Embodiments according to the present disclosure also include methods of treating a patient suffering from an autoimmune disorder, such as a kidney-related autoimmune disorder, including kidney-related diseases, disorders or complications, comprising administering a compound that blocks the interaction of CD40L with CD40 in combination with a compound that blocks the interaction between CD80 and CD28, or administering a therapeutically effective amount of a compound that blocks the interaction of CD40L with CD40 in combination with a compound that blocks the interaction between CD86 and CD28, wherein the kidney-related disease, disorder or complication is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, kidney transplantation, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, kidney infection, nephrolithiasis or IgA nephropathy, by administering to the patient a compound that blocks the interaction of CD40L with CD40. In some embodiments, these methods include administering an anti-CD40L antibody. In some embodiments, these methods include treating patients with autoimmune diseases. Non-limiting examples of autoimmune diseases include systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, and Graves' disease. In some embodiments, the compound that blocks the interaction of CD40L with CD40 is MR1, and the compound that blocks the interaction between CD28 and CD86 or between CD28 and CD80 is abatacept, galiximab, or belatacept. In some embodiments, the compound that blocks the interaction of CD40L with CD40 is 5c8. In some embodiments, these methods include administering a therapeutically effective amount of a combination of MR1 and abatacept or belatacept.In some embodiments, the methods comprise administering a therapeutically effective amount of 5c8 in combination with abatacept or belatacept.

[0141] Kidney-Related Autoimmune Diseases, Disorders and Conditions

[0142] This disclosure relates to treating subjects with one or more autoimmune diseases, including kidney-related autoimmune diseases, disorders and / or conditions. In some embodiments, the autoimmune disease is a kidney-related disease, disorder or condition. In some embodiments, the kidney-related disease, disorder or condition is a complication after kidney transplantation. In some embodiments, the kidney-related disease, disorder or condition is kidney transplant rejection. In some embodiments, the kidney-related disease, disorder or condition is graft-versus-host disease. In some embodiments, the kidney-related disease, disorder or condition is chronic kidney disease. In some embodiments, the kidney-related disease, disorder or condition is nephropathy. In some embodiments, the kidney-related disease, disorder or condition is IgA nephropathy. In some embodiments, the kidney-related disease, disorder or condition is an autoimmune disease of the kidney. In some embodiments, the kidney-related disease, disorder or condition is inflammation or involves inflammation.

[0143] It will be understood that the compounds disclosed herein targeting CD40 or CD40L can be used to combat any disease, disorder or condition in which the kidney is damaged. Non-limiting examples of kidney-related diseases, disorders and conditions include: colitis, drug-induced lupus nephritis, lupus nephritis, nephropathy, chronic kidney disease, graft-versus-host disease, kidney transplantation, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis, IgA nephropathy, systemic Lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, Graves' disease, Abdulhalden-Kaufman-Lignac syndrome (nephrotic cystinosis), abdominal compartment syndrome, acetaminophen-induced nephrotoxicity, acute renal failure / acute kidney injury, acute injury with low back pain after anaerobic exercise (ALPE), renal hypouricemia, acute lobar nephritis (Acute Lobar Nephronia), acute phosphate nephropathy, acute tubular necrosis, adenine phosphoribosyltransferase deficiency, adenoviral nephritis, Alaje syndrome, alkaptonuria, Allport syndrome, Alström syndrome, amyloidosis, ANCA vasculitis associated with endocarditis and other infections, angiomyolipoma, anabolic steroid abuse, analgesic nephropathy, anorexia nervosa and kidney disease, angiotensin antibodies and focal segmental glomerulosclerosis, anticoagulant-associated nephropathy, antiphospholipid syndrome, anti-LRP2 nephropathy, anti-brush border nephropathy, APOL1 mutations in anti-TNF-α therapy-associated glomerulonephritis, manifest mineralocorticoid excess syndrome, aristolochic acid nephropathy, Chinese herbal nephropathy, Balkan endemic nephropathy, urinary arteriovenous malformations and fistulas, atypical hemolytic uremic syndrome, autosomal dominant hypocalcemia, Bard-Biedl syndrome, Bartter syndrome, bath salts and acute kidney injury, beer hyponatremia Potomania), beeturia, beta-thalassemia nephropathy, bile cast nephropathy, Bert-Hoag-Dubbé syndrome, BK polyomavirus nephropathy of native kidney, bladder rupture, bladder sphincter dyssynergia, bladder tamponade, bodybuilder's glomerulopathy, border crosser's nephropathy, Bourbon virus and acute kidney injury, BRASH syndrome, acute renal dysfunction, Byetta and renal failure, C1q nephropathy, C3 glomerulopathy, C4 glomerulopathy, CAKUT (congenital malformation of the kidney and urinary tract), calyceal diverticula, calcineurin inhibitor nephrotoxicity, Callilepsis laurelii poisoning (CallilepsisLaureolaPoisoning), cannabinoid hyperemesis acute renal failure, capillary leak syndrome, carcinoid syndrome and renal failure, cardiorenal syndrome, carfilzomib-induced renal injury, CFHR5 nephropathy, Charcot-Marie-Tooth disease with glomerulopathy, cholesterol emboli, Chargé-Strauss syndrome, chyluria, ciliary disease, cisplatin nephrotoxicity, cold diuresis, colistin nephrotoxicity, collagenous glomerulopathy, collapsing glomerulopathy, collapsing glomerulopathy associated with CMV, combined antiretroviral (cART)-associated nephropathy, congenital adrenal hyperplasia, 17-alpha-hydroxylase / 7 20-lyase deficiency, congenital anomaly of the kidney and urinary tract (CAKUT), congenital nephrotic syndrome, congestive renal failure, conorneal epiphyseal nephrotic syndrome (Conorenalsyndrome) (Mainzer-Saldino syndrome ... Syndrome or Saldino-Mainzer Disease), Contrast Nephropathy, Copper Sulfate Poisoning, Coronavirus (COVID-19)-Associated Renal Failure and Kidney Disease, Cortical Necrosis, Crizotinib-Associated Acute Kidney Injury, Crying Kidneys, Cryocrystalglobulinemia, Cryoglobulinemia, Crystalglobulin-Induced Nephropathy, Crystal Storage Histiocytosis, Cystic Kidney Disease, Acquired Cystic Kidney Disease, Limited Renal Failure, Cystinuria, Cystinosis, Dasatinib-Induced Nephrotic-Range Proteinuria, Deferasirox (Exjade) Nephrotoxicity, Dense Deposition Disease (MPGN) 2), Dent disease (X-linked recessive nephrolithiasis), DHA crystalline nephropathy, dialysis imbalance syndrome, diabetic kidney disease, diffuse mesangial sclerosis, diuresis, mucuna poisoning (Djenkolism), duplicated ureters, EAST syndrome, ectopic kidney, edema, swelling, end-stage renal disease, Edheim-Chester disease, Fabry disease, familial hypocalciuric hypercalcemia, Fanconi syndrome, Fraser syndrome, fibronectin glomerulopathy, fibrillary glomerulonephritis, and immunotactoid glomerulopathy (Immunotactoid glomerulopathy)glomerulopathy), fibrillary glomerulonephritis, congophilic Freiliy syndrome, fluid overload, hypervolemia, focal segmental glomerulosclerosis, focal sclerosis, focal glomerulosclerosis, focal segmental glomerulosclerosis (familial) with complete heart block, Galloway-Mowat syndrome, giant cell (temporal) arteritis with renal involvement, pregnancy-induced hypertension, Gitelman syndrome, glomerular disease, glomerular tubular reflux, glomerulonephritis, diabetes, Goodpasture's syndrome, Green Smoothie Cleanse Nephropathy, HANAC syndrome, HAVANI (ledipasvir and sofosbuvir)-induced kidney injury, hair dye ingestion and acute kidney injury, Hantavirus infection podocytopathy, heat stress nephropathy, hematuria (blood in the urine), hemolytic uremic syndrome (HUS), atypical hemolytic uremic syndrome (aHUS), hemophagocytic syndrome, hemorrhagic cystitis, hemorrhagic fever with renal syndrome (HFRS, Hantavirus nephropathy, Korean hemorrhagic fever, epidemic hemorrhagic fever, epidemic nephropathy (NephropathisEpidemica), hemosiderosis, hemosiderosis associated with paroxysmal nocturnal hemoglobinuria and hemolytic anemia, hepatic glomerulopathy, hepatic veno-occlusive disease, hepatic sinusoidal obstruction syndrome, hepatitis C-associated nephropathy, hepatocyte nuclear factor 1β-associated kidney disease, hepatorenal syndrome, herbal supplements and kidney disease, high altitude renal syndrome, hypertension and kidney disease, HIV-associated immune complex kidney disease (HIVICK), HIV-associated nephropathy (HIVAN), devastating kidney disease, HNF1B-associated autosomal dominant tubulointerstitial kidney disease, horseshoe kidney (renal fusion), Henner's ulcer, hydrophilic polymer emboli, hydroxychloroquine-induced nephrolipidosis, aldosteronism, hypercalcemia, hyperkalemia, hypermagnesemia, hypernatremia, hyperoxaluria, hyperphosphatemia, hypocalcemia, hypocomplementemic urticarial vasculitis syndrome, hypokalemia, hypokalemia-induced renal dysfunction, hypokalemic cycle paralysis, hypomagnesemia, hyponatremia, hypophosphatemia, hypertension, monogenic hypertension, iced tea nephropathy, ifosfamide nephrotoxicity, IgA nephropathy, IgG4 nephropathy, immersion diuresis, immune checkpoint therapy-associated interstitial nephritis, infliximab-associated nephropathy, interstitial cystitis, painful bladder syndrome, interstitial nephritis, megakaryocytic interstitial nephritis, Ivemark's syndrome, JC virus nephropathy, Joubert syndrome, ketamine-associated bladder dysfunction, kidney-based immune disorders, nephrolithiasis, kidney stones, nephrolithiasis due to light chains, Kimura disease, kombucha toxicity, lead nephropathy and lead-related nephrotoxicity, lecithin cholesterol acyltransferase deficiency (LCAT deficiency), leptospirosis nephropathy, light chain deposition disease, monoclonal immunoglobulin deposition disease, light chain proximal tubulopathy, Liddle syndrome, Lightwood-Albright syndromeSyndrome), lipoprotein glomerulopathy, lithium nephrotoxicity, hereditary FSGS due to LMX1B mutations, flank pain and hematuria, lupus, systemic lupus erythematosus, lupus kidney disease, lupus nephritis, lupus nephritis with anti-neutrophil cytoplasmic antibody seropositivity, lupus podocytopathy, lupus-like nephritis, lupus nephritis, Lyme disease-associated glomerulonephritis, lysinuric protein intolerance, lysozyme nephropathy, malarial nephropathy, malignant disease-associated nephropathy, malignant hypertension, soft plaques disease, Marfan syndrome and kidney disease, McKittrick-Wheelock Syndrome, urethral stenosis, medullary cystic kidney disease, uromodulin-associated nephropathy, juvenile hyperuricemia type 1 nephropathy, medullary sponge kidney, megaureter, MELAS syndrome, membranoproliferative glomerulonephritis, membranous nephropathy, membranous nephropathy with microglomeruli, membranous glomerulopathy with occult IgGκ deposition, Mesoameric nephropathy, Metabolic acidosis, metabolic alkalosis, methotrexate-associated renal failure, microscopic polyangiitis, milk-alkali syndrome, minimal change nephropathy, monoclonal gammopathy of renal significance, dysproteinemia, mouthwash toxicity, MUC1 nephropathy, polycystic renal dysplasia, multiple myeloma, myeloproliferative neoplasms and glomerulopathy, nail-patella syndrome, NARP syndrome, nephrocalcinosis, nephrocystin-1 gene deletion and ESRD, nephrogenic systemic fibrosis, nephronophthisis due to nephrocystin-1 gene deletion, renal ptosis (wandering kidney, nephroptosis), nephrotic syndrome, neurogenic bladder, nodular glomerulosclerosis, nongonococcal urethritis, nutcracker syndrome, nephron oligomegaly, orofacial digital syndrome, orotic aciduria, orthostatic hypotension, orthostatic proteinuria, osmotic diuresis, osmotic nephropathy, ovarian hyperstimulation syndrome, oxalate nephropathy, Paget's kidney Kidney), Papillary Necrosis, Papillorenal Syndrome (Renal Coloboma Syndrome, Solitary Kidney Agenesis), Peritoneal-Renal Syndrome, POEMS Syndrome, Posterior Urethral Valve, Podocyte Infolding Glomerulopathy, Postinfectious Glomerulonephritis, Poststreptococcal Glomerulonephritis, Atypical Postinfectious Glomerulonephritis, Postinfectious Glomerulonephritis (IgA Dominant), IgA Nephropathy Mimic, Polyarteritis Nodosa, Posterior Urethral Valve Polycystic Kidney Disease, Post-Obstructive Diuresis, Preeclampsia, Propofol Infusion Syndrome, Proliferative Glomerulonephritis with Monoclonal IgG Deposition (Nasr Disease)Disease), propolis (bee resin)-related renal failure, proteinuria (protein in urine), pseudoaldosteronism, pseudohypobicarbonatemia, pseudohypoparathyroidism, pseudoporphyria, psoriasis and kidney disease, pulmonary-renal syndrome, purple urine bag syndrome, pyelonephritis (kidney infection), suppurative nephropathy, radiation nephropathy, refeeding syndrome, reflux nephropathy, rapidly progressive glomerulonephritis, REN Mutation-induced autosomal dominant tubulointerstitial kidney disease, renal abscess, perinephric abscess, renal agenesis, acute kidney injury associated with renal arcuate vein microthrombosis, renal artery aneurysm, spontaneous renal artery dissection, renal artery stenosis, renal cell carcinoma, renal cyst, renal hypouricemia with exercise-induced acute renal failure, renal infarction, renal osteodystrophy, renal tubular acidosis, renin mutation and autosomal dominant tubulointerstitial kidney disease, renin-secreting tumors (juxtaglomerular cell tumors), osmotic set point reset (Reset Osmostat), retrocaval ureter, retroperitoneal fibrosis, rhabdomyolysis, rhabdomyolysis associated with bariatric surgery, rheumatoid arthritis-associated nephropathy, Rubraca (rucaparib)-associated creatinine increase, sarcoid nephropathy, Saturday night acute kidney injury, schistosomiasis and glomerular disease, Simk immune bone dysplasia, scleroderma renal crisis, serpentine peroneal-polycystic kidney syndrome, Exner syndrome (Exner syndrome) Syndrome), Sezary syndrome, sickle cell nephropathy, Sjögren syndrome and nephropathy, Sri Lankan farmer's kidney disease, carambola nephrotoxicity, renal transplantation, post-hematopoietic cell transplantation renal disease, hepatocyte transplantation-associated renal disease, TAFRO syndrome, tea and toast hyponatremia, tenofovir-induced nephrotoxicity, thin basement membrane disease, benign familial hematuria, thrombotic microangiopathy associated with monoclonal gammopathy, trench nephritis, autosomal dominant thrombotic microangiopathy and CKD, trigoneitis, tuberous sclerosis, tuberous sclerosis complex (TSC), and autosomal dominant nephropathy. autosomal dominant polycystic kidney disease (ADPKD) (TSC2 / PKD1 contiguous gene syndrome), renal tubular dysgenesis, immune complex tubulointerstitial nephritis, tumor lysis syndrome, uremia, uremic optic neuropathy, ureteritis cystica, ureteral prolapse, urethral caruncle, urethral stricture, urinary incontinence, urinary tract infection, urinary tract obstruction, genitourinary fistula, vasomotor nephropathy, vesicoenteric fistula, vesicoureteral reflux, Hippel-Lindau disease, Waldenstrom's macroglobulinemia glomerulonephritis, Wegener's granulomatosis, granulomatosis with polyangiitis, Wunderlich syndrome, Zellweger syndrome, and cerebrohepatorenal syndrome.

[0144] Pharmaceutical compositions and methods of administration

[0145] For treating any of the foregoing disorders, one or more physiologically acceptable carriers can be used to formulate a pharmaceutical composition for use in accordance with the methods of the present disclosure in a conventional manner. Pharmaceutically acceptable carriers are determined in part by the particular composition being administered and by the particular method used to administer the composition. Thus, there are a wide variety of suitable formulations for the compounds useful in the methods of the present disclosure (see, e.g., Remington: The Science and Practice of Pharmacy, 20th Edition, edited by Gennaro et al., Lippincott Williams and Wilkins, 2000).

[0146] Formulations suitable for oral administration include, for example, solid, semi-solid, and liquid systems such as tablets; soft or hard capsules containing multi- or nanoparticles, liquids, or powders; lozenges (including liquid-filled); chewable tablets; gels; rapid-dissolving dosage forms; films; ovules; sprays; and oral / mucoadhesive patches.

[0147] Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic, sterile injection solutions (which may contain antioxidants, buffers, bacteriostatic agents, and solutes that render the formulation isotonic with the blood of the intended recipient), and aqueous and non-aqueous sterile suspensions (which may include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives).

[0148] In accordance with the present disclosure, the compounds can be administered by any suitable means, which can vary depending on the type of disorder being treated and the nature of the compound itself. For example, the compounds can be administered orally, parenterally, or topically. For proteins (such as antibodies), the route of administration preferably includes parenteral, such as intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous. Preferably, parenteral administration is by injection, most preferably intravenous, intramuscular, or subcutaneous injection. The amount to be administered will depend on a variety of factors, such as the clinical symptoms, the weight of the individual, and whether other medications are being administered. It should be understood that the determination of the appropriate dosage form, dose, and route of administration is within the level of ordinary skill in the pharmaceutical and medical arts and is described below.

[0149] In an exemplary pharmaceutical composition containing an anti-CD40L antibody or an antigen-binding fragment thereof, the composition is formulated as a sterile, preservative-free solution of the anti-CD40L antibody or its antigen-binding fragment for intravenous or subcutaneous administration. The formulation can be provided as a disposable prefilled pen, as a disposable (e.g., containing about 1 mL) prefilled glass syringe, or as a disposable mechanism use vial. Preferably, the pharmaceutical composition containing the anti-CD40L antibody or its antigen-binding fragment is transparent and colorless, with a pH of about 5.0 to about 6.9, preferably a pH of about 5.0 to about 6.5, and even more preferably a pH range of about 5.0 to about 6.0. In various embodiments, when reconstituted and administered to a subject, the formulation containing the pharmaceutical composition can contain from about 500 mg to about 1 mg, or from about 400 mg to about 10 mg, or from about 300 mg to about 30 mg, or from about 200 mg to about 50 mg of anti-CD40L antibody or its antigen-binding fragment per mL of solution.

[0150] The administration scheme of selecting therapeutic agent depends on several factors, including the serum or tissue turnover rate of entity, symptom level, immunogenicity of entity and the accessibility of target cells in biological matrix, the general health status of patient, the previous medical history of patient, etc. Preferably, the administration scheme maximizes the amount of therapeutic agent delivered to the patient, consistent with the acceptable side effect level. Therefore, the amount of biological preparation delivered depends in part on the severity of the specific entity and the disease treated. It should be understood that the determination of appropriate dosage form, dosage and administration route is within the level of ordinary technicians in the pharmaceutical field and medical field.

[0151] Dose and exposure level

[0152] The pharmaceutical formulations of the present disclosure may contain from about 0.001 to about 200 mg / kg of an anti-CD40L antibody or antigen-binding fragment thereof, for example, 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 of an anti-CD40L antibody or antigen-binding fragment thereof may be given as a bolus, and the remainder of the antibody dose may be administered by intravenous injection. A predetermined dose of an anti-CD40L antibody or antigen-binding fragment thereof may be administered, for example, over a period of one to two to five hours.

[0153] In other embodiments, a portion of the dose is administered by subcutaneous injection and / or infusion as a bolus, and the remainder of the dose is administered by infusion of the antibody formulation. In some exemplary doses, the antibody formulation may be administered subcutaneously at a dose ranging from about 0.001 to about 200 mg / kg (e.g., 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) of an intravenous injection of an anti-CD40L antibody or antigen-binding fragment thereof. In some embodiments, the dose may be given as a bolus, and the remainder of the antibody dose may be administered by subcutaneous or intravenous injection. A predetermined dose of an anti-CD40L antibody or antigen-binding fragment thereof may be administered, for example, over a period of one hour, or a period of two hours, or a period of three hours, or a period of four hours, or a period of five hours or more.

[0154] Combination therapy

[0155] The antibodies or antibody fragments thereof described herein can be administered alone (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 standard of care treatment, which may or may not include additional therapeutic agents (consisting essentially of the antibody or antibody fragment thereof).

[0156] Adjunctive or combined administration (co-administration) includes simultaneous administration of any antibody or antigen-binding fragment thereof described herein and one or more agents in the same or different dosage forms, or separate administration of the polypeptide and one or more agents (e.g., sequential administration). Such simultaneous or sequential administration preferably results in both the polypeptide and one or more agents being present in the treated patient at the same time.

[0157] Kits and Products

[0158] A test kit containing an antibody or its antigen-binding fragment as described herein and instructions for use is further provided. Test kits typically include a packaged combination of a predetermined amount of reagents and instructions and labels indicating the intended use of the contents of the test kit. The term label or instructions includes any written or recorded material provided on or with the test kit or otherwise accompanying the test kit at any time during the manufacture, transportation, sale or use of the test kit. It can be in a form prescribed by a government agency that manages the manufacture, use or sale of a drug or biological product, and the announcement reflects the agency's approval of manufacture, use or sale for use in humans or veterinary use. Labels or instructions can also cover advertising leaflets and brochures, packaging materials, and audio or video instructions.

[0159] Anti-CD40L antigen binding molecules

[0160] Some embodiments provided herein relate to antibodies that are modified forms of the anti-CD40L antibody hu5c8, comprising a human IgG1 consensus framework having variable light and variable heavy chain CDR sequences of hu5c8, wherein the Fc domain is modified to prevent platelet activation.

[0161] Table 3 provides the references to the present application (and shown in Figure 7-Figure 12 Description of SEQ ID NO).

[0162] Table 3:

[0163]

[0164]

[0165] One embodiment (Embodiment A) is an isolated antibody that binds to CD40L and comprises a light chain and a heavy chain, wherein the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity to SEQ ID NO: 1, and the heavy chain comprises a variable heavy chain region and an Fc region, wherein the heavy chain variable region comprises an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity to SEQ ID NO: 2, and the Fc region comprises an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity to SEQ ID NO: 3. ID NO: 3 having an amino acid sequence with at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity, wherein the Fc region comprises a substitution or a combination of substitutions selected from the group consisting of: C11S, C14S and P23S.

[0166] Another embodiment (embodiment B) is the isolated antibody according to embodiment A, wherein the Fc region further comprises the amino acid substitution C5S.

[0167] In a variation of Examples A and B, the antibody comprises a light chain variable region that does not contain any of the substitutions T33W, S26D and Q27E.

[0168] In other variations of Examples A and B, the light chain variable region comprises the substitution R28K.

[0169] In some variations of Examples A and B, the CDRs of the heavy and light chains have the sequences listed in Table 4.

[0170] Table 4:

[0171] CDR1 light chain ISCRASQRVSSSTYSYMH(SEQ ID NO:15) CDR2 light chain YASNLES (SEQ ID NO: 16) CDR3 light chain QHSWEIPPT (SEQ ID NO: 17) CDR1 heavy chain SYYMY (SEQ ID NO: 18) CDR2 heavy chain EINPSNGDTNFNEKFKS(SEQ ID NO:19) CDR3 heavy chain SDGRNDMDS (SEQ ID NO: 20)

[0172] In yet other variations of embodiments A and B, the light chain variable region comprises the amino acid sequence ICRRASQRVSSSTYSYMH (SEQ ID NO: 15). In still other embodiments, the light chain variable region comprises the amino acid sequence ICRRASQRVSSSTYSYMH (SEQ ID NO: 15) and one or two of the amino acid sequences YASNLES (SEQ ID NO: 16) and QHSWEIPPT (SEQ ID NO: 17).

[0173] In some variations of embodiments A and B, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 1. In yet other embodiments, the light chain variable region consists of the amino acids of SEQ ID NO: 1. In some embodiments, the light chain consists essentially of the amino acid sequence of SEQ ID NO: 7. In other embodiments, the light chain consists of the amino acid sequence of SEQ ID NO: 7. In still other embodiments, the light chain comprises the amino acid sequence of SEQ ID NO: 11. In yet other embodiments, the light chain consists essentially of the amino acid sequence of SEQ ID NO: 11. In still other embodiments, the light chain consists of the amino acid sequence of SEQ ID NO: 11.

[0174] In other variations of embodiments A and B, the antibody comprises a heavy chain variable region that does not contain any of the substitutions T30H, Y33W, or S54N. In some embodiments of the antibodies of embodiments A and B, the light chain variable region does not contain any of the substitutions T33W, S26D, and Q27E. In other variations of embodiments A and B, the light chain variable region does not contain any of the substitutions T33W, S26D, and Q27E, and the heavy chain variable region does not contain any of the substitutions T30H, Y33W, or S54N.

[0175] In yet other variations of embodiments A and B, the heavy chain variable region comprises the substitution K74R. In one embodiment, the heavy chain variable region comprises one or any combination of the amino acid sequences SYYMY (SEQ ID NO: 18), EINPSNGDTNFNEKFKS (SEQ ID NO: 19), and SDGRNDMDS (SEQ ID NO: 20).

[0176] In another embodiment, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 2. In yet another embodiment, the heavy chain variable region consists essentially of the amino acid sequence of SEQ ID NO: 2. In still another embodiment, the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 6. In yet other embodiments, the heavy chain variable region consists essentially of the amino acid sequence of SEQ ID NO: 6. In still other embodiments, the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 6.

[0177] Some embodiments provided herein relate to an isolated antibody, wherein the light chain comprises the amino acid sequence of SEQ ID NO:1 and the heavy chain consists of the amino acid sequence of SEQ ID NO:9.

[0178] Some embodiments provided herein relate to an isolated antibody wherein the light chain consists of the amino acid sequence of SEQ ID NO:7 and the heavy chain consists of the amino acid sequence of SEQ ID NO:9.

[0179] Some embodiments provided herein relate to an isolated antibody, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:5, and the heavy chain consists of the amino acid sequence of SEQ ID NO:9.

[0180] Still another embodiment is an isolated antibody, wherein the light chain consists of the amino acid sequence of SEQ ID NO:11, and the heavy chain consists of the amino acid sequence of SEQ ID NO:9.

[0181] Yet another embodiment is an isolated antibody, wherein the light chain consists of the amino acid sequence of SEQ ID NO:7, and the heavy chain consists of the amino acid sequence of SEQ ID NO:13.

[0182] Another embodiment is an isolated antibody, wherein the light chain consists of the amino acid sequence of SEQ ID NO:11, and the heavy chain consists of the amino acid sequence of SEQ ID NO:13.

[0183] Some embodiments of the present application relate to modifying biomarkers in a subject by treatment with a compound targeting CD40 / CD40L. In some embodiments, the biomarker is a protein involved in CD40 / CD40L activity. In some embodiments, the biomarker is CD40. In some embodiments, the biomarker is CD40L. In some embodiments, the biomarker is NFL. In some embodiments, the biomarker is a pro-inflammatory marker or is associated with inflammation. In some embodiments, the biomarker is a cytokine and / or an interleukin. In some embodiments, the biomarker is one or more of IgA, IgE, CD40L, C3, CXCL9, CXCL10, and / or IgM.

[0184] In some embodiments, treatment with a compound targeting CD40 / CD40L increases the concentration of one or more biomarkers. In some embodiments, treatment with a compound targeting CD40 / CD40L decreases the concentration of one or more biomarkers. In some embodiments, treatment with a compound targeting CD40 / CD40L increases the concentration of at least one biomarker and decreases the concentration of at least one other biomarker. In some embodiments, the baseline level of at least one biomarker serves as a predictor of response to treatment with a compound targeting CD40 / CD40L in a subject with a kidney-related disease, disorder or condition.

[0185] Some of the embodiments provided herein are further described in the following enumerated alternatives.

[0186] 1. A method for predicting the response of a subject with a kidney-related disease or disorder to treatment with a compound directed against CD40L or CD40, the method comprising:

[0187] collecting a sample from the subject;

[0188] measuring the concentration of at least one biomarker present in the sample, wherein the at least one biomarker is CXCL9, CXCL10, IgM, C3, IgA, IgE, CD40L, and CD40;

[0189] wherein the concentration of the at least one biomarker predicts that the subject is likely to respond to treatment of the kidney-related disease or disorder with a compound directed against CD40L or CD40.

[0190] 2. The method of alternative 1, wherein the kidney-related disease or disorder comprises inflammation.

[0191] 3. The method of any of the preceding alternatives, wherein the subject is a mammal and / or a human.

[0192] 4. The method of any of the preceding alternatives, wherein the compound blocks the interaction between CD40 and CD40L.

[0193] 5. The method of any of the preceding alternatives, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

[0194] 6. The method of any of the preceding alternatives, wherein the compound is AT-1501 or 5c8.

[0195] 7. The method of any of the preceding alternatives, wherein the compound is MR1.

[0196] 8. A method as described in any of the preceding alternatives, wherein the kidney-related disease or disorder is selected from the group consisting of: colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, kidney transplantation, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, kidney infection, nephrolithiasis and IgA nephropathy.

[0197] 9. The method of any of the foregoing alternatives, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

[0198] 10. The method of any of the preceding alternatives, wherein the sample is blood, urine, saliva or mucus.

[0199] 11. The method of alternative 10 wherein the sample is urine.

[0200] 12. A method of treating a subject suffering from a kidney-related disease or disorder, the method comprising: administering a therapeutically effective dose of a compound directed against CD40L or CD40.

[0201] 13. The method of Alternative 12 wherein the kidney-related disease or disorder comprises inflammation.

[0202] 14. The method of any one of Alternatives 12-13, wherein the subject is a mammal and / or a human.

[0203] 15. The method of any of Alternatives 12-14, wherein the compound blocks the interaction between CD40 and CD40L.

[0204] 16. The method of any of Alternatives 12-15, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

[0205] 17. The method of any of Alternatives 12-16, wherein the compound is AT-1501.

[0206] 18. The method of any of Alternatives 12-17, wherein the compound is MR1 ​​or 5c8.

[0207] 19. The method of any one of Alternatives 12-18, wherein the compound is administered at least once every three weeks.

[0208] 20. The method of any one of Alternatives 12-19, wherein the compound is administered at a dose of between 200 mg / kg and 100 mg / kg, preferably at a dose of between 1 mg / kg and 20 mg / kg.

[0209] 21. A method as described in any of alternatives 12-20, wherein the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, kidney infection, nephrolithiasis or IgA nephropathy.

[0210] 22. The method of any one of alternatives 12-21, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

[0211] 23. The method of any of Alternatives 12-22 further comprising administering a second pharmaceutically effective compound.

[0212] 24. The method of Alternative Embodiment 23, wherein the second compound blocks the interaction between CD28 and CD86 or between CD28 and CD80.

[0213] 25. The method of Alternative 23 wherein the second compound targets at least one of the biomarkers selected from the group consisting of: an interleukin, a cytokine, and a pro-inflammatory marker.

[0214] 26. The method of Alternative 23 wherein the second compound is CTLA4-Ig fusion protein, abatacept, belatacept, or galiximab.

[0215] 27. The method of any one of Alternatives 12-26, wherein the compound is administered orally, parenterally or topically.

[0216] 28. The method of Alternative 27 wherein the compound is administered parenterally.

[0217] 29. The method of Alternative 28, wherein the compound is administered by injection, most preferably intravenous, intramuscular or subcutaneous injection.

[0218] 30. The method of any of the preceding alternatives, wherein the compound targeting CD40L or CD40 is an antibody comprising:

[0219] (a) a heavy chain variable region (VH), the heavy chain variable region comprising:

[0220] i) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9;

[0221] ii) a CDRH2 domain comprising the sequence shown in SEQ ID NO: 10, 11, 12, 13 or 14; and

[0222] iii) a CDRH3 domain comprising the sequence shown in SEQ ID NO: 15; and

[0223] (b) a light chain variable region (VL), the light chain variable region comprising:

[0224] i) a CDRL1 domain comprising the sequence shown in SEQ ID NO: 16 or 17;

[0225] ii) a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and

[0226] iii) a CDRL3 domain comprising the sequence shown in SEQ ID NO:20.

[0227] 31. The method of any of the preceding alternatives, wherein the compound targeting CD40L or CD40 is an antibody comprising:

[0228] a) a heavy chain variable region (VH), the heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 1, 2, 3 or 4; and

[0229] b) a light chain variable region (VL), which has the amino acid sequence shown in SEQ ID NO: 5, 6, 7 or 8.

[0230] 32. A method of modulating the concentration of at least one biomarker in a subject, the method comprising:

[0231] administering to the subject a therapeutically effective amount of a compound directed against CD40L or CD40,

[0232] Wherein the biomarkers are selected from the group consisting of: IgA, IgE, CD40L, CD40, C3, CXCL9, CXCL10 and IgM.

[0233] 33. The method of Alternative Embodiment 32 wherein the concentration of at least one biomarker increases following administration of the compound.

[0234] 34. The method of Alternative Embodiment 32 wherein the concentration of at least one biomarker decreases following administration of the compound.

[0235] 35. The method of Alternative Embodiment 32 wherein following administration of the compound, the concentration of at least one biomarker increases and the concentration of at least one other biomarker decreases.

[0236] 36. The method of any one of Alternatives 32-35, wherein the subject suffers from a kidney-related disease or disorder.

[0237] 37. The method of Alternative Embodiment 36, wherein the kidney-related disease or disorder comprises inflammation.

[0238] 38. The method of any one of Alternatives 32-37, wherein the subject is a mammal and / or a human.

[0239] 39. The method of any of Alternatives 32-38, wherein the compound blocks the interaction between CD40 and CD40L.

[0240] 40. The method of any of Alternatives 32-39, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

[0241] 41. The method of any of Alternatives 32-40, wherein the compound is AT-1501.

[0242] 42. The method of any one of Alternatives 32-41, wherein the compound is MR1 ​​or 5c8.

[0243] 43. A method as described in any of alternatives 36-42, wherein the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, kidney infection, nephrolithiasis or IgA nephropathy.

[0244] 44. The method of any one of alternatives 36-43, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

[0245] 45. The method of any one of Alternatives 32-44, wherein the compound is administered at least once every three weeks.

[0246] 46. ​​The method of any one of Alternatives 32-45, wherein the compound is administered at a dose of between 200 mg / kg and 100 mg / kg, preferably at a dose of between 1 mg / kg and 20 mg / kg.

[0247] 47. The method of any one of Alternatives 32-46, wherein the compound is administered orally, parenterally or topically.

[0248] 48. The method of Alternative Embodiment 47 wherein the compound is administered parenterally.

[0249] 49. The method of Alternative Embodiment 47, wherein the compound is administered by injection, most preferably intravenous, intramuscular or subcutaneous injection.

[0250] 50. The method of any one of Alternatives 33-49, wherein the compound that targets CD40L or CD40 is an antibody comprising:

[0251] a. a heavy chain variable region (VH), the heavy chain variable region comprising:

[0252] i) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9;

[0253] ii) a CDRH2 domain comprising the sequence shown in SEQ ID NO: 10, 11, 12, 13 or 14; and

[0254] iii) a CDRH3 domain comprising the sequence shown in SEQ ID NO: 15; and

[0255] b. a light chain variable region (VL), the light chain variable region comprising:

[0256] i. a CDRL1 domain comprising a sequence as shown in SEQ ID NO: 16 or 17;

[0257] ii. a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and

[0258] iii. A CDRL3 domain comprising the sequence shown in SEQ ID NO:20.

[0259] 51. The method of any one of Alternatives 32-50, wherein the compound that targets CD40L or CD40 is an antibody comprising:

[0260] a. a heavy chain variable region (VH), the heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 1, 2, 3 or 4; and

[0261] b. a light chain variable region (VL), which has an amino acid sequence as shown in SEQ ID NO: 5, 6, 7 or 8.

[0262] Examples

[0263] Embodiments are further defined in the following examples. It should be understood that these examples are given by way of illustration only. Based on the above discussion and these examples, those skilled in the art can determine the essential characteristics, and without violating its spirit and scope, various changes and modifications of the embodiments can be made to adapt them to various uses and conditions. Therefore, in addition to those modifications shown and described herein, various modifications of the embodiments will become clear to those skilled in the art according to the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. The disclosure of each reference shown herein is incorporated herein by reference in its entirety, and is used for the disclosures cited herein.

[0264] Example 1

[0265] Kidney transplant clinical trials

[0266] Four human subjects with IgAN (Buerger's disease) underwent kidney transplantation and were monitored for a period of 12 weeks. At the end of the twelve weeks, urine samples were collected from each subject. These samples were then screened for changes in proinflammatory markers compared to the levels in each subject's urine before kidney transplantation.

[0267] After 12 weeks, IgAN-related pro-inflammatory markers IgA, IgE, CD40L, and C3 were significantly reduced in patients ( Figures 20A-20D In addition, after 12 weeks, the general transplant pro-inflammatory markers CXCL9, CXCL10, IgM, and C3 decreased ( Figure 21A-Figure 21D ). Considering the robust decrease in CD40L and CD40L-related proteins in subjects following transplantation of healthy kidneys, it appears that subsequent treatment with CD40L-targeting compounds, such as those disclosed herein, may help accelerate recovery and reduction of inflammation in subjects.

[0268] Example 2

[0269] Phase 1b human clinical trial

[0270] The clinical trial was expanded to a 52-week, open-label, single-dose level study with up to 12 participants. Details of the study design are as follows Fig. 22 Participants were administered an immunosuppressive regimen using ATG induction therapy plus CNI-free maintenance therapy (including mycophenolate, corticosteroids, and ticagrelor as an alternative to tacrolimus). Medications were administered as follows:

[0271] rATG: 1.5 mg / kg / dose, intravenously, for 4 doses (total 6 mg / kg). The first dose was initiated before revascularization, and the remaining 3 doses (total 4.5 mg / kg) were given within 5-10 days after transplantation, consistent with the label instructions.

[0272] Corticosteroids: According to standard of care, taper to 5 mg prednisone (or equivalent) per day by day 28 post-transplant.

[0273] MMF: 1000 mg BID, or MPS: 720 mg BID.

[0274] Tigrat: 20 mg / kg via intravenous infusion on days 1, 3, 7, 14, 21, 28, and every 21 days thereafter.

[0275] The first four participants selected for the trial were two women and two men aged 60-77 years ( Fig.23Each of these participants received a kidney transplant and was subsequently dosed as described above. Renal function was monitored using estimated glomerular filtration rate (eGFR) for the next 120 ( Figure 24-25 ).like Fig.25 As shown, ticagliptin was administered at four different times. To date, renal function has been significantly improved with a low hazard ratio.

[0276] As used herein, section headings are for organizational purposes only and should not be construed as limiting the subject matter described in any way. All documents and similar materials cited in this application (including but not limited to patents, patent applications, articles, books, papers and internet web pages) are expressly incorporated by reference in their entirety for any purpose, including the disclosures specifically cited herein. When the definition of a term in the incorporated reference appears to be different from the definition provided in this teaching, the definition provided in this teaching should be used as the standard. It will be understood that there is an implicit "about" before the temperature, concentration, time, etc. discussed in this teaching, so that slight and non-substantial deviations are within the scope of the teaching herein.

[0277] Although the embodiments described herein have been disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the disclosure extends beyond the specifically disclosed embodiments to other alternative embodiments and / or uses of the disclosure and obvious modifications and equivalents thereof. In addition, although several variations of the embodiments have been shown and described in detail, other modifications based on the disclosure (which are within the scope of the disclosure) will be easily clear to those skilled in the art. It is also contemplated that various combinations or sub-combinations may be made to the specific features and aspects of the embodiments, and these combinations or sub-combinations still fall within the scope of the disclosure. It should be understood that the various features and aspects of the disclosed embodiments may be combined or replaced with each other to form different modes or embodiments. Therefore, it is intended that the scope of the disclosure should not be limited by the above-mentioned specifically disclosed embodiments.

[0278] It should be understood, however, that this detailed description, while indicating preferred embodiments, is given by way of illustration only, since various changes and modifications within the spirit and scope of the present disclosure will become apparent to those skilled in the art.

[0279] The terms used in the specification presented herein are not intended to be interpreted in any limited or restrictive manner. Instead, the terms are used only in conjunction with the detailed description of the embodiments of the systems, methods, and related components. In addition, the embodiments may include several novel features, no single feature of which is solely responsible for its desired properties or is considered necessary to practice the embodiments described herein.

Claims

1. A method for predicting the response of a subject with a kidney-related disease or disorder to treatment with a compound directed against CD40L or CD40, the method comprising: collecting a sample from the subject; measuring the concentration of at least one biomarker present in the sample, wherein the at least one biomarker is CXCL9, CXCL10, IgM, C3, IgA, IgE, CD40L, and CD40; wherein the concentration of the at least one biomarker predicts that the subject is likely to respond to treatment of the kidney-related disease or disorder with a compound directed against CD40L or CD40.

2. The method of claim 1, wherein the kidney-related disease or disorder comprises inflammation.

3. The method of claim 1, wherein the subject is a mammal and / or a human.

4. The method of claim 1, wherein the compound blocks the interaction between CD40 and CD40L.

5. The method of claim 1, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

6. The method of claim 1, wherein the compound is AT-1501 or 5c8.

7. The method of claim 1, wherein the compound is MR1.

8. The method of claim 1, wherein the kidney-related disease or disorder is selected from the group consisting of colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, kidney transplantation, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis, and IgA nephropathy.

9. The method of claim 1, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

10. The method of claim 1, wherein the sample is blood, urine, saliva or mucus.

11. The method of claim 10, wherein the sample is urine.

12. A method of treating a subject suffering from a kidney-related disease or disorder, the method comprising: A therapeutically effective dose of a compound directed against CD40L or CD40 is administered.

13. The method of claim 12, wherein the kidney-related disease or disorder comprises inflammation.

14. The method of claim 12, wherein the subject is a mammal and / or a human.

15. The method of claim 12, wherein the compound blocks the interaction between CD40 and CD40L.

16. The method of claim 12, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

17. The method of claim 12, wherein the compound is AT-1501.

18. The method of claim 12, wherein the compound is MR1 ​​or 5c8.

19. The method of claim 12, wherein the compound is administered at least once every three weeks.

20. The method of claim 12, wherein the compound is administered at a dose between 200 mg / kg and 100 mg / kg, preferably at a dose between 1 mg / kg and 20 mg / kg.

21. The method of claim 12, wherein the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis, or IgA nephropathy.

22. The method of claim 12, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

23. The method of claim 12, further comprising administering a second pharmaceutically effective compound.

24. The method of claim 23, wherein the second compound blocks the interaction between CD28 and CD86 or between CD28 and CD80.

25. The method of claim 23, wherein the second compound targets at least one of the biomarkers selected from the group consisting of: an interleukin, a cytokine, and a pro-inflammatory marker.

26. The method of claim 23, wherein the second compound is CTLA4-Ig fusion protein, abatacept, belatacept, or galiximab.

27. The method of claim 12, wherein the compound is administered orally, parenterally, or topically.

28. The method of claim 27, wherein the compound is administered parenterally.

29. The method of claim 28, wherein the compound is administered by injection, most preferably intravenous, intramuscular or subcutaneous injection.

30. The method of claim 12, wherein the compound targeting CD40L or CD40 is an antibody comprising: (a) a heavy chain variable region (VH), the heavy chain variable region comprising: i) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence shown in SEQ ID NO: 10, 11, 12, 13 or 14; and iii) a CDRH3 domain comprising the sequence shown in SEQ ID NO: 15; and (b) a light chain variable region (VL), the light chain variable region comprising: i) a CDRL1 domain comprising the sequence shown in SEQ ID NO: 16 or 17; ii) a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and iii) a CDRL3 domain comprising the sequence shown in SEQ ID NO:

20.

31. The method of claim 12, wherein the compound targeting CD40L or CD40 is an antibody comprising: (a) a heavy chain variable region (VH), the heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 1, 2, 3 or 4; and (b) a light chain variable region (VL), wherein the light chain variable region has the amino acid sequence shown in SEQ ID NO: 5, 6, 7 or 8.

32. A method of modulating the concentration of at least one biomarker in a subject, the method comprising: administering to the subject a therapeutically effective amount of a compound directed against CD40L or CD40, wherein the biomarker is selected from the group consisting of: IgA, IgE, CD40L, CD40, C3, CXCL9, CXCL10 and IgM.

33. The method of claim 32, wherein the concentration of at least one biomarker increases after administration of the compound.

34. The method of claim 32, wherein the concentration of at least one biomarker decreases after administration of the compound.

35. The method of claim 32, wherein after administration of the compound, the concentration of at least one biomarker increases and the concentration of at least one other biomarker decreases.

36. The method of claim 32, wherein the subject suffers from a kidney-related disease or disorder.

37. The method of claim 36, wherein the kidney-related disease or disorder comprises inflammation.

38. The method of claim 32, wherein the subject is a mammal and / or a human.

39. The method of claim 32, wherein the compound blocks the interaction between CD40 and CD40L.

40. The method of claim 32, wherein the compound is an anti-CD40L antibody or an anti-CD40 antibody.

41. The method of claim 32, wherein the compound is AT-1501.

42. The method of claim 32, wherein the compound is MR1 ​​or 5c8.

43. The method of claim 36, wherein the kidney-related disease or disorder is colitis, drug-induced lupus nephritis, lupus nephritis, kidney disease, chronic kidney disease, graft-versus-host disease, immune graft response, transplant rejection, atherosclerosis, diabetic kidney disease, acute kidney injury, renal failure, polycystic kidney disease, hematuria, Fabry disease, lupus kidney disease, Alport syndrome, IgM nephropathy, interstitial nephritis, membranoproliferative glomerulonephritis, simple renal cysts, renal infection, nephrolithiasis, or IgA nephropathy.

44. The method of claim 36, wherein the kidney-related disease or disorder is an autoimmune disease, wherein the autoimmune disease is systemic lupus erythematosus, type 1 diabetes, myasthenia gravis, inflammatory bowel disease, immune thrombocytopenic purpura, rheumatoid arthritis, psoriasis, Addison's disease, Crohn's disease, uveitis, multiple sclerosis, hemolytic anemia, or Graves' disease.

45. The method of claim 32, wherein the compound is administered at least once every three weeks.

46. ​​The method of claim 32, wherein the compound is administered at a dose between 200 mg / kg and 100 mg / kg, preferably at a dose between 1 mg / kg and 20 mg / kg.

47. The method of claim 32, wherein the compound is administered orally, parenterally, or topically.

48. The method of claim 47, wherein the compound is administered parenterally.

49. The method of claim 47, wherein the compound is administered by injection, most preferably intravenous, intramuscular or subcutaneous injection.

50. The method of claim 32, wherein the compound that targets CD40L or CD40 is an antibody comprising: (a) a heavy chain variable region (VH), the heavy chain variable region comprising: i) a CDRH1 domain comprising the sequence shown in SEQ ID NO:9; ii) a CDRH2 domain comprising the sequence shown in SEQ ID NO: 10, 11, 12, 13 or 14; and iii) a CDRH3 domain comprising the sequence shown in SEQ ID NO: 15; and (b) a light chain variable region (VL), the light chain variable region comprising: i) a CDRL1 domain comprising the sequence shown in SEQ ID NO: 16 or 17; ii) a CDRL2 domain comprising the sequence shown in SEQ ID NO: 18 or 19; and iii) a CDRL3 domain comprising the sequence shown in SEQ ID NO:

20.

51. The method of claim 32, wherein the compound that targets CD40L or CD40 is an antibody comprising: (a) a heavy chain variable region (VH), the heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 1, 2, 3 or 4; and (b) a light chain variable region (VL), wherein the light chain variable region has the amino acid sequence shown in SEQ ID NO: 5, 6, 7 or 8.

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