Treatment of lupus using humanized Anti-CXCR5 antibodies

Abstract

To provide methods of preventing or treating lupus in mammals, including humans.SOLUTION: The present invention provides prophylactic, immunotherapeutic, and diagnostic compositions comprising an anti-CXCR5 antibody, and the use of such anti-CXCR5 antibodies in methods of preventing or treating lupus in mammals, including humans.SELECTED DRAWING: None

Description

Technical Field

[0001] The present disclosure relates to anti-CXCR5 antibodies and their use in the amelioration, treatment, or prevention of lupus. The present disclosure also relates to prophylactic, immunotherapeutic, and diagnostic compositions comprising anti-CXCR5 antibodies, and the use of such anti-CXCR5 antibodies in methods for preventing or treating lupus in mammals, including humans.

Background Art

[0002] CXCR5, also known as Burkitt lymphoma receptor 1 (BLR1), CD185, MDR15, and MGC117347, is a G protein-coupled receptor that is a member of the CXC chemokine receptor family. CXCR5 is selectively expressed in recirculating B cells and follicular helper T ( FH ) cells. CXCR5 has a unique ligand, CXC chemokine ligand 13 protein (CXCL13), which is constitutively expressed by stromal cells located in secondary lymphoid organs, B cell follicles of the pleural and peritoneal cavities, and ectopic lymphoid follicles. CXCL13 is one of the most potent B cell chemotactic factors and plays an important role in attracting CXCR5-expressing cells to these regions. The interaction between T cells and B cells in the follicular area leads to B cell proliferation and subsequent B cell maturation, antibody formation, class switching, and the formation of germinal centers (GCs) accompanied by affinity maturation.

[0003] Systemic lupus erythematosus (SLE) is characterized by the pathogenic formation of autoantibodies against nuclear, cytoplasmic, and / or cell surface molecules resulting from B and T cell immunoregulatory deficiencies. The local formation and / or deposition of circulating antigen-antibody immune complexes induces an immune response characterized by remissions and exacerbations, leading to multi-organ system damage and the potential for end-organ failure, which is a factor in a wide range of systemic and organ-specific clinical symptoms.

Summary of the Invention

Means for Solving the Problems

[0004] The present disclosure is a method for treating a patient having lupus, comprising administering to the patient a therapeutically effective amount of an antibody or a fragment thereof that specifically binds to the extracellular domain of human CXCR5, wherein the antibody or the fragment thereof is (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 11 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; (b) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLAS (SEQ ID NO: 59), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (c) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16; (d) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLAS (SEQ ID NO: 64), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (e) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSNLAS (SEQ ID NO: 65), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (f) a variable light chain (V L ) comprising the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, and a variable heavy chain (V H ) comprising the amino acid sequence of SEQ ID NO: 23; (g) a variable light chain comprising the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34; (h) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLA (SEQ ID NO: 66), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (i) The amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLA (SEQ ID NO: 67), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (j) The amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (k) A variable light chain comprising the amino acid sequence of SEQ ID NO: 35 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 37; (l) A variable light chain comprising the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 41, or SEQ ID NO: 43, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 47; (m) A variable light chain comprising the amino acid sequence of SEQ ID NO: 55 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57; or (n) Comprising the amino acid sequences of RSSKSLLHSSGKTYLYW (SEQ ID NO: 69), RMSNLA (SEQ ID NO: 66), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); Provided is a method in which a patient is tested positive for antinuclear antibody at a titer of ≧1:160.

[0005] The present disclosure also provides a method for treating a patient having lupus, the method comprising administering to the patient a therapeutically effective amount of an antibody or a fragment thereof that specifically binds to the extracellular domain of human CXCR5, the antibody or the fragment thereof comprising a variable light chain comprising the amino acid sequence of SEQ ID NO: 32 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33; and the patient is tested positive for antinuclear antibody at a titer of ≧1:160.

[0006] The present disclosure also provides a method of treating a patient having lupus, the method comprising administering to the patient a therapeutically effective amount of an antibody or fragment thereof that specifically binds to the extracellular domain of human CXCR5, the antibody or fragment thereof comprising the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); wherein the patient has been tested positive for antinuclear antibody at a titer of ≧1:160.

[0007] Certain embodiments of the present disclosure will become apparent from the following more detailed description of specific embodiments and the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0008]

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DETAILED DESCRIPTION OF THE INVENTION

[0009] The present disclosure relates to anti-CXCR5 antibodies and their use in the amelioration, treatment, or prevention of lupus. The present disclosure also relates to prophylactic, immunotherapeutic, and diagnostic compositions comprising anti-CXCR5 antibodies, and the use of such anti-CXCR5 antibodies in methods for preventing or treating lupus in mammals, including humans.

[0010] Using standard recombinant DNA methodologies, polynucleotides encoding polypeptides that form the anti-CXCR5 antibodies of the present disclosure are constructed, these polynucleotides are incorporated into recombinant expression vectors, and such vectors are introduced into host cells. See, for example, Green and Sambrook, 2012, MOLECULAR CLONING: A LABORATORY MANUAL (Cold Spring Harbor Laboratory Press, 4th ed.). Enzymatic reactions and purification techniques are performed according to the manufacturer's protocols as commonly accomplished in the art or as described herein. Unless otherwise specified, the nomenclature, as well as the experimental procedures and techniques utilized in connection with analytical chemistry, synthetic organic chemistry, and medicinal chemistry described herein are well known and commonly used in the art. Similarly, conventional techniques are used in chemical synthesis, chemical analysis, pharmaceutical manufacture, formulation, delivery, and patient treatment.

[0011] 1. General Definitions When used in accordance with the present disclosure, the following terms are understood to have the following meanings unless otherwise specified. Unless required by context, the singular terms include the plural and the plural terms include the singular.

[0012] "CXCR5" refers to a naturally occurring, known molecule found on lymphocytes, particularly B cells, and particularly naive B cells; such a molecule isolated from such cells; such a molecule produced recombinantly using known substances and means, as well as nucleic acids encoding CXCR5; and to portions of CXCR5, such as the extracellular (EC) domain that retains features and properties relevant to the practice of the present disclosure, such as CXCL13 binding. Soluble CXCR5 molecules consist essentially of the EC domain of CXCR5 and typically include the first approximately 60 amino acids of the molecule, i.e., the amino-terminal portion of CXCR5.

[0013] CXCR5 is a non-broad host range receptor. CXCL13 is a ligand of CXCR5 and is constitutively expressed on follicular dendritic cells and stromal cells such as lymphoid tissues. CXCL13 specifically attracts a small subset of T cells called B cells and follicular helper T cells (TFH). Furthermore, activated T cells induce or upregulate CXCR5 expression. Thirdly, lymphocyte infiltration into ectopic germinal centers (GCs) has been found to be well correlated with increased disease severity and breakdown of tolerance in certain disorders preset by such abnormal lymph node-like structures. Using in vivo mouse models such as CXCR5− / − and CXCL13− / − mice, the absence of either receptor or ligand results in alterations in GC fine structure by altering T and B cell localization and, if possible, interactions. These mice are also protected against the development of severe collagen-induced arthritis (CIA). When CXCR5 is selectively expressed on mature B cells, this leads to the onset of RA, and blockade of this receptor regulates the arthritis response in affected individuals. Treatment of rheumatoid arthritis with biopharmaceuticals (i.e., anti-TNFα and anti-CD20 antibodies, rituximab) has been shown to be clinically effective; particularly in patients treated with B cell therapy, long-term improvement in clinical signs and symptoms has been shown. Selective targeting of CXCR5, which is expressed only on mature B cells and B helper T cells, does not affect B cell development or the patient's immunodeficiency state. Unlike rituximab, the antibodies disclosed herein are neutralizing antibodies that do not mediate cytotoxicity.

[0014] "CXCR5 disease" is a disease, disorder, illness, condition, abnormality, etc., characterized by, or caused by, overexpression or increased levels of CXCL13 or other CXCR5 ligands, increased levels of B cells, increased levels of B cell activity, increased levels of CXCR5 or inappropriate metabolism and activity of CXCR5.

[0015] The terms "human CXCR5", "hCXCR5", or "hCXCR5 polypeptide" and similar terms refer to the polypeptide disclosed in U.S. Patent No. 8,647,622, which is incorporated herein by reference in its entirety (the terms "polypeptide", "peptide", and "protein" are used interchangeably herein), and are synthesized or isolated from related polypeptides including suitable cell sources and their SNP variants. Related polypeptides include allelic variants (e.g., SNP variants); splice variants; fragments; derivatives; substitution, deletion, and insertion variants; fusion polypeptides; and interspecies homologs, and in some embodiments retain CXCR5 activity and / or are sufficient to generate an anti-CXCR5 immune response. Soluble forms of CXCR5 sufficient to generate an anti-CXCR5 immune response are also included. As will be recognized by those skilled in the art, anti-CXCR5 binding agents such as antibodies can bind to CXCR5 polypeptides, polypeptide fragments, antigens, and / or epitopes when the epitope is part of a larger antigen, part of a larger polypeptide fragment, and further part of a larger polypeptide.

[0016] As used herein, the term "antibody" refers to a protein that can recognize and specifically bind to an antigen. Normal or conventional mammalian antibodies typically consist of two identical pairs of polypeptide chains, each pair consisting of one "light" chain (typically having a molecular weight of about 25 kDa) and one "heavy" chain (typically having a molecular weight of about 50 - 70 kDa). As used herein, the terms "heavy chain" and "light chain" refer to any immunoglobulin polypeptide having a variable domain sequence sufficient to confer specificity to the target antigen. Typically, the amino-terminal portion of each light and heavy chain contains a variable domain of about 100 to 110 or more amino acids typically involved in antigen recognition. The carboxy-terminal portion of each chain typically defines a constant domain involved in effector function. Thus, in a naturally occurring antibody, the full-length heavy chain immunoglobulin polypeptide has a variable domain (V H ) and three constant domains (C H1 , C H2, and C H3 ) and C H1 and C H2 includes a hinge region between, and V H domain is at the amino terminus of the polypeptide, and C H3 domain is at the carboxyl terminus, and the full-length light chain immunoglobulin polypeptide has a variable domain (V L ) and a constant domain (C L ), and V L domain is at the amino terminus of the polypeptide, and C L domain is at the carboxyl terminus.

[0017] Within the full-length light and heavy chains, the variable and constant domains are typically joined by a "D" region of about 10 or more amino acids and a "J" region of about 12 or more amino acids in the heavy chain. The variable regions of each light / heavy chain pair typically form the antigen-binding site. The variable domains of naturally occurring antibodies typically exhibit the same general structure of relatively conserved framework regions (FRs) joined by three hypervariable regions, also called complementarity-determining regions or CDRs. The CDRs from the two chains of each pair are typically aligned by the framework regions to enable binding to a particular epitope. From the amino terminus to the carboxyl terminus, both light and heavy chain variable domains typically include domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0018] As used herein, "anti-CXCR5 antibody" means an antibody or polypeptide (derivative) thereof that specifically binds to human CXCR5 as defined herein, and includes, but is not limited to, a molecule that inhibits or substantially reduces the binding of CXCR5 to its ligand, or inhibits CXCR5 activity.

[0019] One example of an anti-CXCR5 antibody is SAR113244, a potent and specific neutralizing antibody to CXCR5. SAR113244 was generated in an IgG4 framework containing two amino acid substitutions described to reduce half-molecule formation (S241P) and Fc-mediated effector functions (L248E).

[0020] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts.

[0021] The monoclonal antibodies herein specifically include "chimeric" antibodies, where the portions of the heavy and / or light chains are identical or homologous to the corresponding sequences of antibodies derived from a particular species, or belong to a particular antibody class or subclass (type or subtype), and exhibit the desired biological activity of binding to CXCR5, or affect CXCR5 activity or metabolism, and the remaining portions of the chains are identical or homologous to the corresponding sequences of antibodies derived from another species, or belong to another antibody class or subclass, as well as fragments of such antibodies (U.S. Patent No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. U.S.A. 81(21):6851-6855 (1984)). Thus, CDRs from one class of antibodies are transplanted into the FRs of antibodies of a different class or subclass.

[0022] The "humanized" form of a non-human (e.g., murine) antibody is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., F v 、F ab 、F ab’ 、F (ab’)2or other target-binding subsequences of the antibody) and contains sequences derived from non-human immunoglobulins as compared to human antibodies. Generally, humanized antibodies contain substantially all of one, and typically two, variable domains, where all or substantially all of the CDR regions as well as all or substantially all of the FR regions corresponding to those of the non-human immunoglobulin are those of a human immunoglobulin template sequence. Humanized antibodies can also contain at least a portion of the immunoglobulin constant region (F C ), typically those of a selected human immunoglobulin template. Usually, the goal is to have an antibody molecule that is minimally immunogenic in humans. Thus, one or more amino acids of one or more CDRs are also modified to minimize the specific binding function of one or more CDRs to CXCR5 or CXCL13 without substantially minimizing the It is mainly changed to something with low immunogenicity. Instead, the FR can be non-human, but its most immunogenic amino acids are replaced with those having low immunogenicity. Nevertheless, as described above, CDR grafting is not the only way to obtain a humanized antibody. For example, since it is not uncommon for framework residues to play a role in determining the three-dimensional structure of the CDR loops and the overall affinity of the antibody to its ligand, simply modifying the CDR regions is insufficient. Therefore, any means are implemented to modify the non-human parental antibody molecule to make it less immunogenic to humans, and overall sequence identity with a human antibody is not always required. Thus, for example, humanization can be achieved by simply substituting even a few residues, especially those that are exposed on the antibody molecule, not buried within the molecule, and thus not readily accessible to the host immune system. Such methods are taught herein with respect to the substitution of "mobile" or "flexible" residues of the antibody molecule, and the aim is to reduce or attenuate the immunogenicity of the resulting molecule without including the specificity of the antibody to its epitope or determinant. See, for example, Studnicka et al., Protein Eng. 7(6):805-14 (1994); Lazar et al., Mol. Immunol. 44(6):1986-98 (2007); Sims et al., J. Immunol. 151(4):2296-308 (1993); Chothia et al., J. Mol. Biol. 196(4):901-17 (1987); Carter et al., Proc. Natl. Acad. Sci. U.S.A. 89(10):4285-89 (1992); Presta et al., J. Immunol. 151(5):2623-32 (1993), International Application No. 2006 / 042333 and U.S. Patent No. 5,869,619.

[0023] Antibodies are humanized by a variety of techniques including CDR grafting (European Publication No. EP0239400; International Publication No. WO91 / 09967; and U.S. Patents Nos. 5,530,101 and 5,585,089), veneering and resurfacing (European Publication Nos. EP0592106 and EP0519596; Padlan, Mol. Immunol. 28(4-5):489-98 (1991); Studnicka et al., Protein Eng. 7(6):805-14 (1994); and Roguska et al., Proc. Natl. Acad. Sci. U.S.A. 91(3):969-73 (1994)) and chain shuffling (U.S. Patent No. 5,565,332). Human antibodies are made by a variety of methods known in the art including, but not limited to, phage display methods, see U.S. Patents Nos. 4,444,887; 4,716,111; 5,545,806; and 5,814,318; and International Publications WO98 / 46645, WO98 / 50433, WO98 / 24893, WO98 / 16654, WO96 / 34096, WO96 / 33735, and WO91 / 10741, using transgenic animals such as rodents, using chimeric cells, etc.

[0024] The term "antibody fragment" refers to intact or full-length chains or antibodies, usually portions of the target binding or variable regions. Examples of antibody fragments include, but are not limited to, F ab 、F ab’ 、F (ab’)2 and F v fragments. A "functional fragment" or "analog of an anti-CXCR5 antibody" is one that can bind to a ligand or interfere with or substantially reduce the ability of a receptor to initiate signal transduction. As used herein, a functional fragment is usually synonymous with "antibody fragment" and, with respect to an antibody, an F v 、F ab 、F (ab’)2 and the like that can bind to a ligand or interfere with or substantially reduce the ability of a receptor to initiate signal transduction. "Fv The "fragment" consists of a dimer of one heavy chain and one light chain variable domain by non-covalent bond (V H -V L dimer). In addition, the three CDRs of each variable domain interact. When it is an intact antibody, V H -V L dimer defines the target binding site on the surface. In summary, the six CDRs confer target binding specificity to the intact antibody. However, even a single variable domain (or half of the F containing only three C v DRs specific to the target) can have the ability to recognize and bind the target.

[0025] "Single-chain F v ", "sF v " or "scAb" antibody fragments contain the V H and V L domains of the antibody, and these domains exist in a single polypeptide chain. Usually, the F v polypeptide further contains a polypeptide linker, which is often a flexible molecule between the V H and V L domains, and can form the desired structure for target binding in sFv.

[0026] The F ab fragment contains the variable and constant domains of the light chain and the variable and first constant domain (C H1 ) of the heavy chain. The F ab’ fragment is different from the F H1 fragment by the addition of some residues at the carboxyl terminus of the C ab domain so as to include one or more cysteines from the antibody hinge region. The F ab’ fragment is produced by cleavage of the disulfide bond at the hinge cysteine of the F (ab’)2 pepsin digestion product. Further enzymatic and chemical treatments of the antibody can produce other functional fragments of interest.

[0027] As used herein, the term "antigen" or "target antigen" refers to a molecule or portion of a molecule that can be bound by an antibody of the present disclosure and can be used in an animal to produce an antibody that can further bind to an epitope of the antigen. A target antigen can have one or more epitopes. For each target antigen recognized by an antibody, the antibody can compete with a native antibody that recognizes the target antigen.

[0028] As used herein, the term "epitope" refers to the region of an antigen that is bound by an antibody of the present disclosure. An antibody is said to specifically bind to an antigen if it preferentially recognizes its antigen target in a complex mixture of proteins and / or macromolecules. As used herein, the term "specifically binds" refers to the ability of an antibody to bind to an antigen containing an epitope having a Kd of at least about 1×10 -6 M, 1×10 -7 M, 1×10 -8 M, 1×10 -9 M, 1×10 -10 M, 1×10 -11 M, 1×10 -12 M or higher and / or to an epitope having an affinity at least 2-fold higher than its affinity for a non-specific antigen.

[0029] As used herein, the term "antigen-binding site" refers to the site of an antibody of the present disclosure to which an antigen or epitope binds. The antigen-binding site of an antibody is typically described with reference to the CDRs of the antibody.

[0030] The phrase "substantially identical" with respect to an antibody chain polypeptide sequence is interpreted to mean an antibody chain that exhibits at least 70%, 80%, 90%, 95%, or more sequence identity with a reference polypeptide sequence. The term with respect to a nucleic acid sequence is interpreted to mean a nucleotide sequence that exhibits at least about 85%, 90%, 95%, 97% or more sequence identity with a reference nucleic acid sequence.

[0031] The terms "identity" or "homology" mean the percentage of nucleotide bases or amino acid residues of a candidate sequence that are identical to the residues of the corresponding sequence being compared after aligning the sequences, introducing gaps if necessary to achieve the maximum percent identity of the entire sequences, and without considering any conservative substitutions as part of sequence identity. Either an N-terminal or C-terminal extension, or an insertion, is considered to reduce identity or homology. Methods of alignment and computer programs are available and well known in the art. Sequence identity is measured using sequence analysis software.

[0032] The terms "antibody" or "antigen", and terms such as "functional fragment", "variant", "derivative or analog", and their forms, are compounds or molecules having a qualitative biological activity common to the full-length antibody or antigen of interest. For example, a functional fragment or analog of an anti-CXCR5 antibody is one that can bind to the CXCR5 molecule, or one that can interfere with or substantially reduce the ability of a ligand such as CXCL13, or is an agonist or antagonist antibody that binds to CXCR5. An example is an scFv molecule. With respect to CXCR5, a variant or derivative thereof that is not identical to native CXCR5 but is used for the purposes of the present disclosure is a molecule that, for example, is not identical to wild-type CXCR5 but is used as an immunogen that gives rise to an antibody that selectively binds to wild-type CXCR5.

[0033] A "substitution" variant has at least one amino acid residue in the native sequence removed and replaced by a different amino acid insertion at the same position in that location. The substitution can be single, where only one amino acid of the molecule is replaced, or multiple, where two or more amino acids are replaced within the same molecule. Multiple substitutions are at contiguous sites. Also, one amino acid can be replaced by multiple residues, in which case such a variant includes both substitution and insertion. An "insertion" variant has one or more amino acids inserted immediately adjacent to an amino acid at a specific position in the native sequence. By immediately adjacent to an amino acid is meant being attached to either the α-carboxyl or α-amino functional group of the amino acid. A "deletion" variant has one or more amino acids of the native amino acid sequence removed. Usually, a deletion variant has one or two amino acids deleted in a specific region of the molecule.

[0034] "Antibody homolog" or "homolog" refers to any molecule that specifically binds to CXCR5 as taught herein. Thus, an antibody homolog can be a native or recombinant antibody, either modified or unmodified, and includes portions of an antibody that retain a desired biological property such as binding to CXCR5, e.g., F ab or F v molecules, single-chain antibodies, polypeptides carrying one or more CDR regions, etc. The amino acid sequence of the homolog need not be identical to that of a naturally occurring antibody, but can be altered or modified to carry substituted amino acids, inserted amino acids, deleted amino acids, amino acids other than the 20 normally found in proteins, etc., to obtain a polypeptide with enhanced or other beneficial properties.

[0035] An antibody having a homologous sequence is an antibody having an amino acid sequence with sequence homology to the amino acid sequence of the CXCR5 antibodies of the present disclosure. Preferably, the homology is based on the amino acid sequence of the variable region of the antibodies of the present disclosure. When applied to the amino acid sequences herein, for example, Pearson et al., Proc. Natl. Acad. Sci. U.S.A. 85(8): 2444-2448 (1988), "sequence homology" is defined as a sequence having at least about 90%, 91%, 92%, 93%, 94% or more sequence homology to another amino acid sequence, and more preferably, a sequence having at least about 95%, 96%, 97%, 98%, or 99% sequence homology, as determined by the FASTA search method.

[0036] The term "functional equivalent" includes antibodies, antibody homologs, chimeric antibodies, artificial antibodies and modified antibodies having a homologous sequence. For example, each functional equivalent is defined by the ability to bind to CXCR5, the ability to inhibit CXCR5 signal transduction or function, or the inhibition of the binding of CXCL13 and other ligands to CXCR5. Those skilled in the art will understand that there is an overlap between the group of molecules called "antibody fragments" and the group called "functional equivalents". Methods for producing functional equivalents that retain the CXCR5 binding ability are known to those skilled in the art and are disclosed, for example, in International Publications WO93 / 21319 and WO89 / 09622, and European Publications EP0 239,400, EP0 338,745, and EP0 332,424.

[0037] An "isolated" or "purified" antibody substantially does not contain other contaminating proteins from the cellular material or the cell or tissue source or medium from which the protein is derived, and when chemically synthesized, substantially does not contain chemical precursors or other chemical substances. "Substantially free of cellular material" The term "substantially free" includes the preparation of antibodies in which the polypeptide / protein is separated from the cellular components of the cells from which it is isolated or recombinantly produced. Thus, antibodies substantially free of cellular material include preparations of antibodies in which the contaminating protein is less than about 30%, 20%, 10%, 5%, 2.5% or 1% (by dry weight). When the antibody is recombinantly produced, it is preferably substantially free of culture medium, i.e., the culture medium represents a volume less than about 20%, 10%, 5%, 2.5%, or 1% of the protein preparation. When the antibody is produced by chemical synthesis, it is preferably substantially free of chemical precursors or other chemicals, and reagents, i.e., the antibody of interest is separated from the chemical precursors or other chemicals involved in the synthesis of the protein. Thus, such a preparation of the antibody includes chemical precursors or compounds other than the antibody of interest that are less than about 30%, 20%, 10%, 5%, or 1% (by dry weight). In one embodiment of the disclosure, the antibody is isolated or purified.

[0038] "Antagonist" refers to a molecule that can inhibit one or more biological activities of a target molecule, such as signal transduction by CXCR5. Antagonists can interfere with the binding of a receptor to its ligand by disabling or killing cells activated by the ligand and / or by interfering with signal transduction after receptor or ligand activation (e.g., tyrosine kinase activation) or ligand binding to the receptor, and vice versa. Antagonists can completely block receptor-ligand interactions and substantially reduce such interactions. All such aspects of intervention by antagonists are considered equivalent for the purposes of this disclosure. Accordingly, within the scope of this disclosure are antagonists (e.g., neutralizing antibodies) that bind to CXCR5, CXCL13, or other ligands of CXCR5, or complexes of CXCR5 and its ligands, such as CXCL13; amino acid sequence variants or derivatives of CXCR5 or CXCL13 that antagonize the interaction between CXCR5 and a ligand, such as CXCL13; soluble CXCR5 optionally fused to a heterologous molecule such as an immunoglobulin region (e.g., an immunoadhesin); complexes comprising CXCR5 associated with another receptor or biomolecule; synthetic or natural sequence peptides that bind to CXCR5; etc.

[0039] "Agonist" refers to a compound including proteins, polypeptides, peptides, antibodies, antibody fragments, conjugates, macromolecules, small molecules, and activates one or more biological activities of CXCR5. The agonist binds to and interacts with the ligand of the receptor, and vice versa, by acting as a mitogen for cells activated by the ligand and / or interfering by cell inactivation or signal transduction inhibition after the ligand binds to the receptor. All such aspects of the intervention by the agonist are considered equivalent for the purposes of the present invention. Accordingly, within the scope of the present disclosure are agonists that bind to CXCR5, CXCL13, or other ligands of CXCR5, or complexes of CXCR5 and its ligands, such as CXCL13; amino acid sequence variants or derivatives of CXCR5 or CXCL13 that promote the interaction between CXCR5 and a ligand, such as CXCL13; soluble CXCR5 optionally fused to a heterologous molecule, such as an immunoglobulin region (e.g., immunoadhesin); complexes containing CXCR5 associated with another receptor or biomolecule; synthetic or natural sequence peptides that bind to CXCR5; and the like. An agonist is generally an entity that directly activates CXCR5, for example, to convey a signal.

[0040] The terms "cell", "cell line", and "cell culture" include their progeny. It is also understood that all progeny may not be precisely identical in, for example, DNA content due to deliberate or inadvertent mutations. Variant progeny having the same biological properties of the same function or purpose are included when screening the originating cells. A "host cell" as used in the present disclosure is usually a prokaryotic or eukaryotic host selected as a design choice.

[0041] "Transformation" of a cellular organism, cell, or cell line having a nucleic acid means introducing the nucleic acid into a target cell such that the nucleic acid is replicable as an extrachromosomal element or by chromosomal integration, and optionally by expression. "Transfection" of a cell or organism by a nucleic acid refers to the uptake of the nucleic acid, e.g., an expression vector, by the cell or organism, regardless of whether any of the coding sequences are actually expressed. The terms "transfected host cell" and "transformed" refer to a cell into which a nucleic acid has been introduced. Typical prokaryotic host cells include various strains of Escherichia coli (E. coli). Typical eukaryotic host cells are mammalian cells such as Chinese hamster ovary, or cells of human origin. The introduced nucleic acid sequence may be from the same species as the host cell or a different species from the host cell, or may be a hybrid nucleic acid sequence containing some foreign and some homologous nucleic acids. Transformation can also occur by transduction or infection with virus-derived elements.

[0042] The term "vector" means a nucleic acid construct, carrier, containing a nucleic acid, transgene, foreign gene, or gene of interest operably linked to suitable control sequences for the expression of the introduced gene in a suitable host. Such control sequences include, for example, a promoter that acts in transcription, any optional operator sequences that control such transcription, a sequence encoding a suitable mRNA ribosome binding site, and sequences that control the termination of transcription and translation. The vector may be a plasmid, phage particle, or simply a potential genomic insertion. Once transformed into a suitable host, the vector can replicate and function independently of the host genome and, in some instances, is integrated into the host cell genome. In this specification, when a plasmid is the form of the vector generally used, the terms "plasmid" and "vector" are used interchangeably. However, the present disclosure is intended to include other forms of vectors known in the art, such as, for example, viruses, synthetic molecules carrying nucleic acids, liposomes, etc., and those that provide an equivalent carrier function that is known or will become known.

[0043] For the purposes of treatment, "mammal" refers to any animal classified as a mammal, including humans, farm animals, non-human primates, and zoo, sports or pet animals such as dogs, horses, cats, cows, etc.

[0044] As used herein, the term "patient" includes human and animal subjects.

[0045] "Disorder" is any condition that would benefit from treatment with the antibodies of the present disclosure. "Disorder" and "condition" are used interchangeably herein and include chronic and acute disorders or diseases, including those pathologies that predispose a patient to the disorder in question.

[0046] The term "lupus" as used herein refers to all types and manifestations of lupus. For example, manifestations of lupus include systemic lupus erythematosus; lupus nephritis; skin manifestations (e.g., cutaneous lupus erythematosus, such as manifestations seen in skin lesions or rashes); CNS lupus; cardiovascular, pulmonary, hepatic, hematological, gastrointestinal and musculoskeletal manifestations; neonatal lupus erythematosus; childhood systemic lupus erythematosus, drug-induced lupus erythematosus; antiphospholipid syndrome; and complement deficiency syndromes that result in lupus manifestations.

[0047] As used herein, the term "treatment" or "treating" refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those having a disorder as well as those in which a disorder is likely to occur or in which a disorder is to be prevented. In certain embodiments, the antibodies are used for the treatment of lupus.

[0048] As used herein, the term "pharmaceutical composition" or "therapeutic composition" refers to a compound or composition that, when appropriately administered to a patient, can induce the desired therapeutic effect. One embodiment of the present disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of at least one antibody of the present disclosure.

[0049] As used herein, the term "pharmaceutically acceptable carrier" or "physiologically acceptable carrier" refers to one or more pharmaceutical substances suitable for achieving or enhancing the delivery of one or more antibodies of the present disclosure.

[0050] When used in reference to a pharmaceutical composition comprising one or more antibodies of the present disclosure, the terms "effective amount" and "therapeutically effective amount" refer to an amount or dosage sufficient to produce the desired therapeutic result. More specifically, a therapeutically effective amount is an amount of antibody sufficient to inhibit one or more of the clinically defined pathological processes associated with the condition being treated over a period of time. The effective amount will vary depending on the particular antibody used and will also depend on various factors and conditions regarding the patient being treated, as well as the severity of the disorder. For example, when the antibody is administered in vivo, factors such as age, weight, and the health of the patient, as well as the dose-response curve and toxicity data obtained from preclinical animal experiments, are among the factors considered. Determination of the effective amount or therapeutically effective amount of a given pharmaceutical composition is within the ability of one of ordinary skill in the art.

[0051] 2. CXCR5 Antibody In some embodiments, the antibody is an isolated antibody or a fragment thereof that specifically binds to the extracellular domain of human CXCR5. The antibody or fragment thereof is: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 11 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; (b) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLAS (SEQ ID NO: 59), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (c) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16; (d) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLAS (SEQ ID NO: 64), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (e) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSNLAS (SEQ ID NO: 65), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (f) a variable light chain (VL) comprising the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21 and a variable heavy chain (VH) comprising the amino acid sequence of SEQ ID NO: 23; (g) a variable light chain comprising the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34; (h) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLA (SEQ ID NO: 66), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (i) the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLA (SEQ ID NO: 67), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63);(j) The amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63); (k) A variable light chain containing the amino acid sequence of SEQ ID NO: 35 and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 37; (l) A variable light chain containing the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 41, or SEQ ID NO: 43, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 47; (m) A variable light chain containing the amino acid sequence of SEQ ID NO: 55, and SEQ ID NO: 56 or SEQ ID NO: 57; A variable heavy chain containing the amino acid sequence of; or (n) It can contain the amino acid sequences of RSSKSLLHSSGKTYLYW (SEQ ID NO: 69), RMSNLA (SEQ ID NO: 66), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63).

[0052] In another embodiment, the antibody or fragment thereof comprises a variable light chain containing the amino acid sequence of SEQ ID NO: 32 and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 33. In another embodiment, the antibody or fragment thereof comprises a variable light chain containing the amino acid sequence of SEQ ID NO: 70 and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 33.

[0053] In another embodiment, the antibody or fragment thereof contains the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), MQHLEYPYT (SEQ ID NO: 60), GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63).

[0054] The antibody or fragment thereof may further comprise one or more constant regions. One or more constant region domains are C H1 、C H2 、C H3 、and / or C LOne or more of the constant regions are derived from an IgG antibody, which may be, for example, an IgG4 antibody. The antibody or fragment thereof may be a single chain Fv.

[0055] Antibodies of the present disclosure are also described or specified in terms of cross-reactivity. Also included in the present disclosure are antibodies that bind CXCR5 polypeptides having at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, and at least 50% identity to CXCR5 (as calculated using methods known in the art and described herein).

[0056] The antibodies of the present disclosure are also described or specified in terms of their binding affinity to CXCR5 of interest. -7 Lower than M, about 10 -6 Lower than M or about 10 -5 K is lower than M D It can be combined with about 10 -8 About 10 -15 M, about 10 -8 About 10 -12 M, about 10 -9 About 10 -11 M, or about 10 -8 About 10 -10 Equilibrium dissociation constant of M or K D Higher binding affinity of the antibody of interest is beneficial, such as one having: ##STR00013## The present disclosure also provides antibodies that competitively inhibit the binding of an antibody to an epitope of the present disclosure as determined by any method known in the art for determining competitive binding, such as the immunoassays described herein. In some embodiments, the antibody competitively inhibits binding to the epitope by at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, or at least 50%.

[0057] A variant antibody or mutant, or mutein, is one in which one or more amino acid residues are altered in, for example, one or more of the hypervariable regions of the antibody. Alternatively, or in addition, one or more alterations (e.g., substitutions) of framework residues are introduced into the antibody, resulting in an improvement in the binding affinity of the antibody variant for CXCR5. Examples of framework region residues that are altered include those that bind the antigen directly, non-covalently (Amit et al., Science 233(4765):747-753 (1986)); those that interact with / affect the structure of the CDR (Chothia et al., J. Mol. Biol. 196(4):901-917 (1987)); and / or those that are L -V H related to the interface (European Publication No. EP0239400). In certain embodiments, one or more alterations of such framework region residues result in an enhanced binding affinity of the antibody for cognate antigen. For example, from about one to about five framework residues are altered in the embodiments of the present disclosure. Sometimes, even when hypervariable region residues are not altered this is sufficient to produce antibody variants suitable for use in preclinical trials. However, typically, antibody variants may include alterations in one or more of the hypervariable regions. The constant region may also be altered to obtain desirable or more desirable effector properties.

[0058] The residues of the hypervariable regions that are altered are altered randomly, and especially when the starting binding affinity of the parental antibody is such as that of the randomly generated antibody variants, the altered binding in the assay is readily screened as taught herein.

[0059] One procedure for obtaining antibody variants, such as CDR variants, is "alanine scanning mutagenesis" (Cunningham et al., Science 244(4908):1081-1085 (1989)). One or more residues in the hypervariable region are replaced with alanine or polyalanine residues. Those hypervariable region residues that demonstrate functional sensitivity to the substitution are then improved by introducing additional or other mutations at the position or site of the substitution. Thus, the sites for introducing diverse amino acid sequences are predetermined, but the nature of the variants themselves need not be predetermined. Similar substitutions are attempted with other amino acids depending on the desired properties of the residues being scanned.

[0060] A more systematic method for identifying amino acid residues to modify involves identifying hypervariable region residues involved in the step of binding CXCR5 and hypervariable region residues that are little or not at all involved in CXCR5 binding. For each ala mutant tested for enhanced binding to CXCR5, an alanine scan of the unbound hypervariable region residues is performed. In another embodiment, those residues that are significantly involved in the binding of CXCR5 are selected and modified. The modification includes deletion of the residue or insertion of one or more residues adjacent to the residue of interest. However, usually the modification involves substitution of the residue with another amino acid. Conservative substitutions are the first substitutions. If such substitutions result in a change in biological activity (e.g., binding affinity), then another conservative substitution is made to determine whether a more substantial change is obtained.

[0061] Even more substantial modifications in the presentation of antibody scope and biological properties are achieved by the step of selecting amino acids, and usually the properties from the resident sites are more substantially different. Thus, such substitutions are made while maintaining: (a) the structure of the polypeptide backbone in the region of the substitution, for example as a sheet or helical structure; (b) the change or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.

[0062] For example, naturally occurring amino acids have common side chain properties: (1) Hydrophobic: Methionine (M or met), Alanine (A or ala), Valine (V or val), Leucine (L or leu), and Isoleucine (I or ile); (2) Neutral, hydrophilic: Cysteine (C or cys), Serine (S or ser), Threonine (T or thr), Asparagine (N or asn), and Glutamine (Q or gln); (3) Acidic: Aspartic acid (D or asp) and Glutamic acid (E or glu); (4) Basic: Histidine (H or his), Lysine (K or lys), and Arginine (R or arg); (5) Residues affecting the chain direction: Glycine (G or gly) and Proline (P or pro), and (6) Aromatic: Tryptophan (W or trp), Tyrosine (Y or tyr), and Phenylalanine (F or phe) are grouped based on this.

[0063] Non-conservative substitutions can involve the step of exchanging an amino acid for an amino acid from another group. Conservative substitutions can involve the exchange of an amino acid for another amino acid within one group of amino acids.

[0064] Preferred amino acid substitutions are: (1) those that reduce susceptibility to proteolysis, (2) those that reduce susceptibility to oxidation, (3) those that alter binding affinity, and (4) those that confer or modify other physicochemical or functional properties of such analogs. Analogs can include mutants of various sequences in addition to naturally occurring peptide sequences. For example, one or more amino acid substitutions (preferably conservative amino acid substitutions) are made to naturally occurring sequences (preferably portions of the polypeptide outside the domain) that form intermolecular contacts. Conservative amino acid substitutions should not substantially change the structural characteristics of the parent sequence without a change in the bulk or structure of the R group or side chain (e.g., the substituted amino acid should not tend to disrupt helices that occur in the parent sequence or other types of secondary structures that characterize the parent sequence) "Proteins, Structures and "Molecular Principles" (Creighton, ed., W. H. Freeman and Company, New York (1984)); "Introduction to Protein Structure" (Branden & Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et al., Nature 354(6349):105 - 06 (1991).

[0065] Typically, an antibody variant having improved biological properties has an amino acid sequence that has at least 75% amino acid sequence identity or similarity with the amino acid sequence of either the heavy or light chain variable domain of a parental anti - human CXCR5 antibody, and is often at least 80%, at least 85%, at least 90%, and at least 95% identical. Identity or similarity to the parental antibody sequence is defined herein as the percentage of amino acid residues of a candidate sequence that are identical (i.e., the same residue) or similar (i.e., amino acid residues from the same group based on common side - chain properties as described above) to the parental antibody residues after aligning the sequences and introducing gaps, if necessary, to achieve maximum percent sequence identity.

[0066] Alternatively, the antibody variant is the heavy and light chains of an anti - CXCR5 antibody, or F cGenerated by systematic variation of the FR and CDR regions of the domain. Another procedure for generating antibody variants involves the use of affinity maturation using phage display (Hawkins et al., J. Mol. Biol. 226(3):889-96 (1992), and Lowman et al., Biochemistry 30(45):10832-38 (1991)). Bacteriophage coat protein fusions (Smith, Science 228(4705):1315-17 (1985); Scott et al., Science 249(4967):386-90 (1990); Cwirla et al., Proc. Natl. Acad. Sci. U.S.A. 87(16):6378-82 (1990); Devlin et al., Science 249(4967):404-06 (1990); and U.S. Patent No. 5,223,409) are known to be useful for associating the phenotype of the presented protein or peptide with the genotype of the bacteriophage particles encoding them. The F ab domain of the antibody is also presented on the phage (McCafferty et al., Nature 348(6301):552-54 (1990); Barbas et al., Proc. Natl. Acad. Sci. U.S.A. 88(18):7978-82 (1991); and Garrard et al., Biotechnology (NY) 9(12):1373-77 (1991)).

[0067] Monovalent phage display consists of presenting a set of protein variants as fusions to the bacteriophage coat protein of the phage particle (Bass et al., Proteins 8(4):309-14 (1990)). Affinity maturation of various proteins, or improvement of the equilibrium binding affinity, can be achieved, for example, by focusing on the CDR regions of the antibody (Barbas et al., Proc. Natl. Acad. Sci. U.S.A. 91(9):3809-13 (1994); and Yang et et al., J. Mol. Biol. 254(3): 392-403 (1995)), achieved previously by successful application of mutagenesis, monovalent phage display, and functional analysis (Lowman et al., J. Mol. Biol. 234(3): 564-78 (1993); and U.S. Patent No. 5,534,617).

[0068] Many (e.g., 10 6 or more) libraries of protein variants that differ at defined positions in the sequence are constructed in bacteriophage particles, each of which contains DNA encoding a specific protein variant. After cycles of affinity purification using immobilized antigen, individual bacteriophage clones are isolated and the amino acid sequence of the displayed protein is deduced from the DNA.

[0069] After production of the antibody variant, the biological activity of that molecule against the parental antibody is determined as taught herein. It may include determination of binding affinity and / or other biological activities or physical properties of the antibody. In one embodiment, a panel of antibody variants is prepared and the binding affinities for the antigen are screened. One or more antibody variants selected from the screening are optionally subjected to one or more further biological activity assays to confirm that the antibody variant has new or improved properties. In other embodiments, the antibody variant retains the ability to bind CXCR5 with a binding affinity similar to or better / higher than that of the parental antibody.

[0070] Antibody variants so selected are subjected to further modification, often depending on the intended use of the antibody. Such modifications can include further changes in the amino acid sequence, fusion to heterologous polypeptides, and / or covalent modifications. For example, cysteine residues not involved in maintaining the correct structure of the antibody variant are usually replaced by serine to improve the oxidative stability of the molecule and prevent abnormal cross-linking. Conversely, cysteine is added to the antibody to improve stability (especially when the antibody is an antibody fragment such as an F v fragment).

[0071] Functional equivalents are produced by exchanging different CDRs of different antibody chains within a composite FR derived from a framework or multiple antibodies. Thus, for example, different classes of antibodies can produce different CXCR5 antibody types and isotypes with a given set of CDRs by substitution of different heavy chains, such as IgG 1-4 , IgM, IgA 1-2 or IgD. Similarly, artificial antibodies within the scope of the present disclosure are produced by embedding a given set of CDRs within a completely synthetic framework.

[0072] CDRs are typically important for epitope recognition and antibody binding. However, changes are made to the residues containing the CDRs without interfering with the ability of the antibody to recognize and bind to homologous epitopes. For example, changes are made that increase the binding affinity of the antibody to the epitope without affecting epitope recognition. Some studies have examined the effect of introducing one or more amino acid changes at various positions in the antibody sequence based on knowledge of the primary antibody sequence and its properties, such as levels of binding and expression (Yang et al., J. Mol. Biol. 254(3):392-403 (1995); Rader et al., Proc. Natl. Acad. Sci. U.S.A. 95(15):8910-15 (1998); and Vaughan et al., Nat. Biotechnol. 16(6):535-39 (1998)).

[0073] Accordingly, equivalents of the desired antibody can be generated by altering the sequences of the heavy and light chain genes in CDR1, CDR2 or CDR3, or the framework regions, using methods such as oligonucleotide-mediated site-directed mutagenesis, cassette mutagenesis, error-prone PCR, DNA shuffling, or mutagenic strains of Escherichia coli (Vaughan et al., Nat. Biotechnol. 16(6):535-39 (1998); and Adey et al., 1996, Chap. 16, pp. 277-91, in "Phage Display of Peptides and Proteins", eds. Kay et al., Academic Press). Methods for altering the nucleic acid sequence of a primary antibody result in antibodies with improved affinity (Gram et al., Proc. Natl. Acad. Sci. U.S.A. 89(8):3576-80 (1992); Boder et al., Proc. Natl. Acad. Sci. U.S.A. 97(20):10701-05 (2000); Davies et al., Immunotechnology 2(3):169-79 (1996); Thompson et al., J. Mol. Biol. 256(1):77-88 (1996); Short et al., J. Biol. Chem. 277(19):16365-70 (2002); and Furukawa et al., J. Biol. Chem. 276(29):27622-28 (2001)).

[0074] To determine whether a particular antibody homolog binds to human CXCR5, any conventional binding assay can be used. Useful CXCR5 binding assays include FACS analysis, ELISA assays, radioimmunoassays, etc., and detect the binding of an antibody to human CXCR5 and the resulting function. The full-length and soluble forms of human CXCR5 taught herein are useful in such assays. Binding of an antibody or homolog to CXCR5, or a soluble fragment thereof, is detected conventionally by use of a secondary antibody specific for the immunoglobulin of the species from which the antibody or homolog is derived.

[0075] To determine whether a particular antibody or homolog significantly blocks the binding of CXCL13 or other ligand to human CXCR5, any suitable competitive assay is used. Useful assays include, for example, ELISA assays, FACS assays, radioimmunoassays, etc., and quantify the ability of an antibody or homolog that competes with CXCL13 or other ligand for binding to human CXCR5. Preferably, the ability of a ligand to block the binding of labeled human CXCR5 to an immobilized antibody or homolog is measured.

[0076] The ability of an antibody or homolog to bind to human CXCR5 is evaluated by testing its ability to bind to + human CXCR5 + cells. Suitable CXCR5

[0077] cells for use in determining whether a particular antibody or homolog binds to human CXCR5 are mammalian tissue culture cells transformed with DNA encoding full-length human CXCR5 and expressing CXCR5 on the cell surface, or are of the B cell lineage. + Binding of an antibody or homolog to the cells is detected by staining the cells with a fluorescently labeled secondary antibody specific for the same species of immunoglobulin from which the test antibody homolog is derived. A fluorescence-activated cell sorter (“FACS”) is used to detect and quantify any binding. See generally Shapiro, “Practical Flow Cytometry,” Alan R. Liss, Inc., New York, N.Y. (1985).

[0078] Also, the ability of an antibody homolog to block the binding of a ligand such as CXCL13 to human CXCR5 is determined by pre-incubating excess ligand with + CXCR5 cells and quantifying the extent to which the bound ligand blocks the binding of the antibody or homolog to the cells. +Binding to cells is quantified by FACS analysis using a fluorescently labeled secondary antibody specific for the same species of immunoglobulin from which the antibody homolog being tested is derived. Alternatively, competitive assays are formed using a labeled ligand or antibody known in the art.

[0079] Antibody fragments that recognize specific epitopes are generated by known techniques. Traditionally, these fragments have been derived from proteolytic digestion of intact antibodies (see, for example, Morimoto et al., J. Biochem. Biophys. Methods 24(102):107-17 (1992); and Brennan et al., Science 229(4708):81-83 (1985)). For example, the F ab and F (ab’)2 fragments are produced by proteolytic cleavage of immunoglobulin molecules using enzymes such as papain (which produces the F ab fragment) or pepsin (which produces the F (ab’)2 fragment). The F (ab’)2 fragment contains the variable region, the light chain constant region and the constant region C H1 domain of the heavy chain. However, these fragments are produced directly by recombinant host cells. For example, antibody fragments are isolated from antibody phage libraries. Alternatively, F (ab’)2 -SH fragments are recovered directly from E. coli and chemically linked to form F(ab’) 2 fragments (Carter et al., Biotechnology (NY) 10(2):163-67 (1992)). According to another approach, F (ab’)2 fragments are isolated directly from recombinant host cell cultures. Other techniques for the production of antibody fragments will be apparent to those skilled in the art. In other embodiments, the selected antibody is a single-chain F v fragment (F V ) (International Publication No. WO93 / 16185).

[0080] 3. Antibody Therapeutic Compositions and Their Administration A therapeutic or pharmaceutical composition comprising one or more antibodies of the present disclosure for the treatment of lupus is within the scope of the present disclosure. Such a therapeutic or pharmaceutical composition can comprise a therapeutically effective amount of an antibody-drug conjugate in admixture with a pharmaceutically or physiologically acceptable formulation selected for compatibility with the mode of administration.

[0081] Acceptable pharmaceutical substances are preferably non-toxic to the recipient at the dosages and concentrations employed.

[0082] The pharmaceutical composition can contain formulation substances for modifying, maintaining, or preserving, for example, the pH, osmolality, viscosity, transparency, color, isotonicity, odor, sterility, stability, degradation or release rate, absorption, or penetration of the composition. Suitable formulation substances include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine), antibacterial agents, antioxidants (such as ascorbic acid, sodium sulfite, or sodium bisulfite), buffers (such as borate, bicarbonate, Tris-HCl, citrate, phosphate, or other organic acids), fillers (such as mannitol or glycine), chelating agents (such as ethylenediaminetetraacetic acid (EDTA)), complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin), fillers, monosaccharides, disaccharides, and other carbohydrates (such as glucose, mannose, or dextrin), proteins (such as serum albumin, gelatin, or immunoglobulins), coloring agents, fragrances and diluents, emulsifiers, hydrophilic polymers (such as polyvinylpyrrolidone), low molecular weight polypeptides, salt-forming counterions (such as sodium), preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide), solvents (such as glycerin, propylene glycol, or polyethylene glycol), sugar alcohols (such as mannitol or sorbitol), suspending agents, surfactants or wetting agents (such as pluronic; PEG; sorbitan esters; polysorbates such as polysorbate 20 or polysorbate 80; triton; tromethamine Tamin; Lecithin; Cholesterol or tyloxapal; Stability enhancers (such as sucrose or sorbitol), isotonicity enhancers (such as alkali metal halides - preferably sodium chloride or potassium chloride - or mannitol sorbitol), delivery vehicles, diluents, excipients and / or pharmaceutical adjuvants may be mentioned (for example, REMINGTON’S PHARMACEUTICAL SCIENCES (18th Ed., A.R.Gennaro, ed., Mack Publishing Company 1990), and subsequent editions, which are incorporated herein by reference for any purpose).

[0083] Optimal pharmaceutical compositions will be determined by those skilled in the art, for example, depending on the intended route of administration, delivery format, and desired dosage. Such compositions can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the antibodies of the present disclosure.

[0084] The main vehicle or carrier in the pharmaceutical composition can be virtually either aqueous or non-aqueous. For example, suitable vehicles or carriers for injection can be water, physiological saline solution, or artificial cerebrospinal fluid, and it is also possible that other substances common in parenteral administration compositions are supplemented. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. Other exemplary pharmaceutical compositions include Tris buffer at about pH 7.0 - 8.5, or acetate buffer at about pH 4.0 - 5.5, and can further include sorbitol or suitable substitutes. In one embodiment of the present disclosure, the antibody composition is prepared for storage in the form of a lyophilized cake or aqueous solution by mixing a selected composition having the desired purity with an optional formulation. Further, using suitable excipients such as sucrose, the antibody is formulated as a lyophilized product.

[0085] The pharmaceutical compositions of the present disclosure are selected for parenteral delivery. Alternatively, the compositions are selected for delivery by inhalation or via the gastrointestinal tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the scope of the art.

[0086] The formulation ingredients are present at concentrations acceptable for the site of administration. For example, buffers are used to maintain the composition at physiological pH or slightly lower pH, typically within a pH range of about 5 to about 8.

[0087] When parenteral administration is contemplated, the therapeutic compositions for use in the present disclosure can be in the form of a pyrogen-free, parenterally acceptable aqueous solution containing the desired antibody in a pharmaceutically acceptable vehicle. A particularly suitable vehicle for parenteral injection is sterile distilled water formulated as a sterile isotonic solution in which the antibodies of the present disclosure are properly preserved. Yet another preparation can include formulations of antibodies with agents that provide for the controlled or sustained release of the product delivered via depot injection, such as injectable microspheres, biodegradable particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads or liposomes. Hyaluronic acid can also be used, which can have the effect of promoting the duration of circulation. Other suitable means for introducing the antibody include implantable drug delivery devices.

[0088] In one embodiment, the pharmaceutical composition is formulated for inhalation. For example, the antibody is formulated as a dry powder for inhalation. Inhalation solutions containing the antibody are also formulated with a high-pressure gas for aerosol delivery. In yet another embodiment, the solution is nebulized.

[0089] Certain formulations are also contemplated for oral administration. In one embodiment of the present disclosure, antibodies administered in this manner are formulated with or without carriers customarily used in the synthesis of solid dosage forms such as tablets and capsules. For example, capsules are bioavailability When the "i" is maximized and pre-systemic degradation is minimized, it is designed to release the active portion of the formulation at the location of the gastrointestinal tract. Additional agents are included to facilitate the absorption of the antibody. Diluents, flavoring agents, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrants and binders are also used.

[0090] Another pharmaceutical composition can comprise an effective amount of an antibody in a mixture with a non-toxic excipient suitable for the manufacture of tablets. By dissolving the tablets in sterile water or another suitable vehicle, a solution is prepared in unit dosage form. Suitable excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate or bicarbonate, lactose or calcium phosphate; or binders such as starch, gelatin or acacia; or lubricants such as magnesium stearate, stearic acid or talc.

[0091] Further pharmaceutical compositions of the present disclosure comprising formulations containing an antibody in sustained or controlled release formulations will be apparent to those skilled in the art. Various other sustained or controlled release means, such as liposome carriers, biodegradable microparticles, or techniques for formulating porous beads and depot injections are also known to those skilled in the art. Further examples of sustained release preparations include semipermeable polymer matrices in the form of shaped articles, such as films or microcapsules. Sustained release matrices can include polyesters, hydrogels, polylactide, copolymers of L-glutamic acid and gamma-ethyl-L-glutamic acid, poly(2-hydroxyethyl methacrylate), ethylene vinyl acetate, or poly-D(-)-3-hydroxybutyric acid. Sustained release compositions also include liposomes prepared by any of several methods known in the art.

[0092] The pharmaceutical compositions of the present disclosure for in vivo administration should generally be sterile. This is achieved by filtration through a sterile filtration membrane. If the composition is lyophilized, sterilization using this method is carried out either before or after lyophilization and reconstitution. Compositions for parenteral administration are stored in lyophilized form or in solution. Additionally, parenteral compositions are usually placed in a container having a sterile access port with a stopper penetrable by a hypodermic needle, such as an intravenous solution bag or vial.

[0093] When the pharmaceutical composition is formulated, it is stored in a sterile vial as a solution, suspension, gel, emulsion, solid, or anhydrous or lyophilized powder. Such formulations are stored either in a ready-to-use form or in a form that needs to be reconstituted before administration (e.g., lyophilized).

[0094] The present disclosure also encompasses kits for producing single-dose units. The kits can each contain both a first container having a dry protein and a second container having an aqueous formulation. Single and multi-chamber filled syringes (e.g., kits containing liquid syringes and lyophilized syringes) are also included within the scope of the present disclosure.

[0095] The effective amount of the pharmaceutical composition containing the antibody used for treatment depends, for example, on the treatment situation and purpose. Those skilled in the art thus understand that the appropriate dosage levels for a treatment vary in part depending on the molecule being delivered, the indication for which the antibody is used, the route of administration, as well as the size (body weight, body surface area or organ size) and condition (age and general health status) of the patient. Thus, the clinician can titrate the dosage and modify the route of administration to obtain an optimal therapeutic effect. Typical dosages can range from about 200 mg to 500 mg.

[0096] The frequency of administration depends on the pharmacokinetic parameters of the antibody in the formulation used. Typically, the clinician administers the composition until the dosage achieves the desired effect. Thus, The composition is administered as a single dose, as two or more doses over time (which may or may not contain the same amount of the desired molecule), or as a continuous infusion via an implant device or catheter. Further refinement of suitable dosages is routinely made by those skilled in the art and is thus within the scope of activities routinely carried out. Appropriate dosages are confirmed via the use of appropriate dose-response data.

[0097] The route of administration of the pharmaceutical composition is in accordance with known methods, for example, orally; via injection by subcutaneous, intravenous, intraperitoneal, intracerebral (intraparenchymal), intraventricular, intramuscular, intraocular, intraarterial, intraportal or intralesional routes; via sustained release systems; or by implant devices. If desired, the composition is administered by bolus injection, or by continuous infusion, or by implant device.

[0098] The composition is also administered locally via the implantation of a membrane, sponge or other suitable material in which the desired molecule is absorbed or encapsulated. When an implant device is used, the device is implanted in any suitable tissue or organ and delivery of the desired molecule is effected via diffusion, timed release bolus or continuous administration.

[0099] The following examples are illustrative of specific embodiments of the present disclosure and various uses thereof. They are described for illustrative purposes only and should not be construed as limiting the scope of the present disclosure in any way.

Examples

[0100] Clinical trial protocol Description of all tests and design plans This is a single-site, double-blind, randomized, placebo-controlled trial of sequential escalating doses of SAR113244 in patients with lupus.

[0101] The total period per patient from screening to the end-of-study (EOS) visit is up to 20 weeks, · Screening: within 4 weeks. · Observation phase from the first administration of the investigational drug to the final assessment: 112 days including two treatment days within 4 weeks (i.e., 84 days from the final dose). · Visit on the 113th day (EOS). · Post - study observation: Day 226 for anti - drug antibody (ADA) assessment (only for patients positive for ADA at EOS) including.

[0102] The dose escalation is designed to proceed from the initial dose to the highest dose until the occurrence of related events that support the determination of the maximum tolerated dose. For example, · The number of adverse events (AEs) with a sensitivity / intensity high enough to make a meaningful determination · The proportion of the same type of AEs high enough to make a determination.

[0103] The dose escalation was stopped as soon as an impact on patient protection or an unacceptable risk was identified or predicted. The severity rate was adapted to SLE patients taking into account event characteristics: type, likelihood of harm, occurrence, progression, and monitorability. The occurrence of unique serious AEs was not in itself a criterion for decision - making (assessment was based on the nature of the reported events).

[0104] The decision to progress from dose "n" to the next - higher "n + 1" dosing level group was based on a blinded safety report: in dosing - level cohort "n" containing at least one placebo in cohort 1, at least 6 out of 8 patients (cohort 1) or 12 out of 16 patients for at least 28 days after the second administration.

[0105] Discussion of the study design and selection of the control group This double - blind, randomized, placebo - controlled SC multiple - ascending - dose study design is well - established for phase I dose - escalation trials and was considered appropriate for evaluating safety, tolerability, and preliminary pharmacokinetics (PK) and pharmacodynamics (PD) (Buoen et al., J. Clin. Pharmacol. 45(10):1123 - 1136 (2005)).

[0106] Patients were hospitalized for 2 days at each SAR113244 administration (i.e., from the evening before the administration day through the next day). The administration of the investigational drug to each patient in Cohort 1 was staggered such that only 1 patient per week was administered during the first 2 weeks. The third patient in the group was not administered earlier than the next week (the 3rd week), which was at least 1 day before administering the rest of the group, i.e., 1:1:1:5. Additionally, patients were randomized such that only 1 out of the first 4 patients received a placebo. The administration was not staggered in Cohort 2.

[0107] Selection of the Test Population Patients were included in the study according to the following criteria. Eligibility Criteria Demographics I01. Male or female patients, aged 18 to 75 years inclusive. I02. Body mass index ≥18.0 and <35.0 kg / m 2 . Health Status I03. Meet the American College of Rheumatology or Systemic Lupus Erythematosus International Collaborating Clinics Classification Criteria for SLE for at least 6 months. I04. Patients with mild to moderate active SLE: The Safety of Estrogens in Lupus Erythematosus National Assessment-Systemic Lupus Erythematosus Disease Activity Index (SELENA-SLEDAI) score comprehensively includes the range of 2 - 9. I05. Female patients who use a highly effective contraceptive method for pregnancy regulation approved by the health authorities for at least 3 months after the last dose and have a negative pregnancy test result. Only included if the patient does not wish or is unable to be tested for pregnancy, is postmenopausal for longer than 12 months or sterilized for longer than 3 months. For male patients with a potentially pregnant partner (including lactating women), use a dual contraceptive method according to the following algorithm for up to 2 months after the last dose: (condom) plus (intrauterine device or hormonal contraception). I06. Presence of at least one of the following serum markers of autoantibody production in screening: · Anti-nuclear antibody (ANA) ≥ 1:160 · Anti-double-stranded (ds) DNA antibody ≥ 2 × the lowest positive level. Control I07. Informed consent was documented before any test-related procedures were performed. I08. Applicable and / or subject to the application of the health insurance system in accordance with the recommendations of national laws effective for biomedical research. I09. Not under the administration of any administrative or legal regulations. Exclusion criteria Medical history and clinical status E01. Unstable or severe symptoms of SLE based on the clinical judgment at the time of entry, in the opinion of the treating physician, require the initiation or change to baseline of concomitant medications, especially high-dose glucocorticoids and / or immunosuppressive medications contraindicated (see E22 criteria) during a series of tests or hospitalization within 30 days before screening, except for minor surgery. E02. Active or chronic, severe neuropsychiatric lupus: · Uncontrolled seizure disorder. · Acute confusional state (delirium) or organic mental disorder. · Psychosis. E03. Severe active lupus nephritis or chronic renal insufficiency: · Require current treatment with cyclophosphamide. · Activated sediment in urine test (erythrocyte casts, hematuria due to SLE). · Urine protein:creatinine ratio > 2 mg / mL. · Estimated creatinine clearance ≤ 50 mL / min or serum creatinine ≥ 1.5 mg / dL (132.6 μmol / L) or serum albumin < 3.2 g / dL in two consecutive estimations at least two weeks apart. E04. Patients receiving erythropoietin for the treatment of anemia. E05. Antiphospholipid syndrome: History of antiphospholipid antibody syndrome and use of oral anticoagulants or antiplatelet agents (excluding acetylsalicylic acid ≤ 325 mg / day), or history of arterial or venous thrombosis within 12 months of screening in patients. E06. Any recent signs of sudden bleeding due to thrombocytopenia within 28 days of screening or during the screening period. E07. In the opinion of the principal investigator of the clinical trial, the patient's health or satisfactory living conditions are at risk, or test results (e.g., uncontrolled hypertension, uncontrolled diabetes), age-related comorbidities, especially congestive heart failure, acute or active heart disorders (e.g., acute myocardial infarction, myocarditis), chronic heart conditions, poorly controlled or progressive (e.g., documented left / right ventricular hypertrophy, clinically significant valvular disease, poorly controlled hypertension, cardiomyopathy), chronic obstructive pulmonary disease, frequent asthma exacerbations that complicate the interpretation, SLE-related clinical problems or other coexisting conditions. E08. Previous or current history of malignant tumors within 5 years prior to screening (excluding carcinomas in situ of the cervix, non-metastatic squamous or basal cell carcinomas that have been adequately treated > 1 year prior to screening). E09. History of congenital or primary immunodeficiency. E10. Conditions other than SLE that require oral treatment with corticosteroids. The use of topical (for conditions such as contact dermatitis) or corticosteroid inhalation (for conditions such as asthma) is not excluded. E11. Marked baseline prolongation of the QT / QTc interval, e.g., repeated demonstration of a QTc interval > 450 ms. E12. History of substance abuse, drug dependence or alcoholism with remission lasting < 1 year prior to screening. Biological status E13. Positive results for any of the following tests: hepatitis B surface antigen (HBsAg), anti-hepatitis B core antibody, anti-hepatitis C virus antibody, anti-human immunodeficiency virus (HIV) 1 and 2 antibodies. E14. Infection: · Positive IgM antibody titer in the presence of a negative IgG titer against Epstein-Barr virus, cytomegalovirus (CMV), or hepatitis A virus. · Current or recent (within 4 weeks of screening) signs or symptoms of infection, excluding minor infections, onychomycosis, or vaginal candidiasis fungal infections. · Hospitalization or active infection requiring intravenous (IV) antibodies within the past 60 days or treatment with oral antibiotics for 30 days prior to screening. · Patients with a history of chronic infection or any frequency of recurrent infection that is considered unacceptable at the discretion of the principal investigator of the clinical trial. · Any history or evidence of opportunistic infection within 6 months of screening, including severe CMV. · History of recurrent herpes zoster, active herpes zoster / varicella within 3 months of screening, varicella exposure within 21 days before screening and / or herpes encephalitis, eye · History of severe herpes infections such as ocular herpes, disseminated herpes. · Frequency of recurrence of oral or genital herpes simplex lesions (≧6 / year). · Patients with latent (or latent, history of treatment) or active tuberculosis (TB) regardless of treatment history as defined below · Any signs or symptoms suggesting active TB in the treatment history or clinical examination, · Patients with a positive QuantiFERON TB Gold test. This test is repeated once if it is considered indeterminate or false positive, and before being included, it is confirmed that the patient has a negative result. · Chest X-rays within 3 months before eligible visit are consistent with before TB infection or Mycobacterium TB exposure, including but not limited to apical scars, apical fibrosis, or multiple calcium-containing granulomas. This did not include non-caseating granulomas. The need for chest X-rays was evaluated according to guidelines and local practices based on the patient's risk of TB. · Patients with a history of TB are enrolled in the trial if adequate treatment is described by an expert, and the patient must be negative for the QuantiFERON TB Gold test. · Patients with a close relationship with humans with active TB. · Patients treated with granulocyte colony-stimulating factor due to neutropenia. It is known or considered that the patient is allergic to E15.SAR113244 or the excipient. E16. Receive any vaccine within 3 months before randomization (baseline) visit. History of severe allergic or anaphylactic reactions to any biologic agent. E17. Patients with the following abnormal test findings · Hemoglobin < 8.5 g / dL (< 85 g / L). · White blood cells < 2000 / mm 3 (< 2.0×10 9 / L). · Neutrophils < 1000 / mm 3 (< 1.0×10 9 / L). · Absolute lymphocyte count < 800 / mm 3 (< 0.8×10 9 / L). · B cells < 50 / mm 3 and T cells < 500 / mm 3 . · Platelets < 50,000 / mm 3 (< 50.0×10 9 / L). · Alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST) > 1.5 times the upper limit of normal (ULN). · Alkaline phosphatase > 1.5 ULN. · Immunoglobulins (IgG, IgM, IgA) below the lower limit of normal (LLN). E18. Positive results in urine drug screening (amphetamine / methamphetamine, psychostimulants, benzodiazepines, cannabinoids, cocaine, opioids) unless medically prescribed in combination with other medications. E19. Positive urine alcohol (ethanol) test Interfering drugs Oral treatment with corticosteroids (>0.3 mg / kg / day prednisone or equivalent) within 30 days before the first administration of the investigational medicinal product, excluding periods of ≤3 days from 30 to 15 days before administration. · Intra-articular steroid injections within 30 days before the first administration E21. It is not preferred to continue a stable baseline corticosteroid dose from screening until at least 6 weeks after the final dose administration. Single-dose adjustment of prednisone (or equivalent) up to ±5 mg / day was possible. E22. Immunomodulatory / immunosuppressive treatments (excluding corticosteroids): · Any B-cell depleting agent (rituximab, belimumab or other investigational medicinal product) that does not demonstrate a return of CD19+ B cells to >50 / μL within either 6 months of randomization or 5 half-lives of the biologic, whichever is longer. · Abatacept within 3 months before randomization; other biologic therapies (e.g., anti-tumor necrosis factor α, anti-interleukin 6) at either 30 days before randomization to the trial or 5 half-lives of the biologic, whichever is longer. · Initiation of methotrexate, azathioprine, hydroxychloroquine or other anti-malarial drugs (chloroquine, quinacrine), mycophenolate mofetil or dapsone within 90 days before administration: · The following doses were exclusive: methotrexate >25 mg / week; azathioprine >2.5 mg / kg / day; hydroxychloroquine >400 mg / day; chloroquine, >3.5 mg / kg / day, quinacrine >100 mg / day; dapsone >200 mg / day; mycophenolate mofetil >2.5 g / day and initiated <90 days before randomization or <14 days before trial screening and with dose fluctuations during the screening period. · Administration of cyclosporine, tacrolimus, sirolimus, thalidomide, lenalidomide, IV Ig and / or plasma exchange therapy administered within 3 months before screening. · Administration of leflunomide within 6 months before screening unless a formal drug withdrawal procedure was administered and documentation provided. · Receive cyclophosphamide (IV or oral) or any other alkylating agent within 6 months of screening. E23. Non-steroidal anti-inflammatory drugs (including COX-2) at variable doses within 14 days before screening and during the screening period. E24. Use of warfarin or heparin. E25. Concurrent participation or involvement within 4 months before screening or 5 half-lives (whichever is longer) in any other trial involving the investigational drug or any other experimental procedure. General condition E26. Pregnancy (defined as a positive pregnancy test), lactating. E27. Any patient judged by the principal investigator of the trial to be non-compliant during the trial and who is considered unable to cooperate due to language problems or mental retardation. E28. The patient is an employee of the clinical site, such as the principal investigator of the trial or any sub-investigator, research assistant, pharmacist, trial coordinator, other staff or related person; the patient or related person is an employee of the sponsor.

[0108] Removal of the patient from treatment or assessment Each patient could withdraw from the trial at any time and for any reason if the patient so decided. The patient could also withdraw at the discretion of the principal investigator of the trial. All withdrawals from the trial procedures were recorded in the electronic case report form (eCRF). If possible, the patient was evaluated using the EOS procedure, including PK samples if appropriate.

[0109] Patients who could not continue treatment due to AE were followed according to the trial procedures until at least 84 days after the last injection, up to the scheduled date of trial completion (EOS visit), or until recovery or stabilization of any AE, whichever came last.

[0110] For any patient who could not be returned to the site, the principal investigator of the clinical trial made all efforts to contact the patient (e.g., contacting the patient's family or primary care physician and rechecking available registrations or health management databases) to determine his / her health status, including at least his / her vital status. Attempts to contact the patient were recorded in the patient's record (e.g., the time and date of attempted telephone contact, receipt certificates for registered letters sent).

[0111] Patients who withdrew from the trial were not re-registered for the trial. Their inclusion and treatment numbers were not reused.

[0112] Treatment Treatment administered SAR1133244 The injectable solution of SAR1133244 was sterile, non-pyrogenic, injectable, colorless to slightly yellow colored, used as a 100 mg / mL solution for SC administration, and placed in 2R ISO glass vials with elastomeric closures.

[0113] Each vial contained nominally 150 mg (1.5 mL) of SAR113244 (0.2 mL in excess). The pH of the solution was between 5.5 and 6.5. The solution contained the following excipients: water for injection, sucrose, arginine hydrochloride, sodium citrate, sodium chloride, polysorbate 20, and hydrochloric acid / sodium hydroxide solution for pH adjustment if necessary.

[0114] Placebo · Isotonic saline (0.9%) injection solution in vial · As required by the manufacturer, the need for specific operations. The investigational medicinal product (IMP; SAR113244 or placebo) was administered as an SC injection into the abdomen. Patients fasted for at least 10 hours before administration and for 2 hours after administration.

[0115] For administration that requires multiple injections, the injections were alternated between the upper left and upper right quadrants and the lower left and lower right quadrants of Zone 4, 10 cm from the navel; injections on the same day were administered in different quadrants.

[0116] Identity of the investigational drug The SAR113244 100 mg / mL vials were placed in cartons as an open-label supply. The labeled content conformed to local control specifications and requirements.

[0117] SAR113244 100 mg / mL was stored between 2°C and 8°C (36°F and 48°F), protected from light, and with limited agitation (no rotation). SAR113244 was not frozen. Light exposure was permitted during preparation and administration.

[0118] Isotonic saline (0.9%) was stored according to the manufacturer's requirements.

[0119] The batch numbers of SAR113244 and placebo were C1037128 and 14384011, respectively.

[0120] Method of assigning patients to treatment groups Patients who met all eligibility criteria and signed the informed consent were assigned a patient number and a randomization number. Randomization was performed by a centralized randomization procedure using an interactive response system.

[0121] For the 500 mg cohort, randomization was stratified according to plasmablast / plasma cell levels (< or ≥ 5% of total B cells) using a 1:1 ratio at screening.

[0122] Exchangeable patients were assigned a different identification number (i.e., 500 + the number of the exchanged patient). Each patient received the same treatment and treatment sequence as the withdrawn patient.

[0123] Selection of doses in the trial Based on the blinded safety and preliminary PK results of study TDU11406 in healthy subjects with up to 500 mg of SAR113244 or placebo, the doses to be repeatedly administered to SLE patients in this study were determined to be 250 mg and 500 mg once every 4 weeks (Q4W) as two injections. The higher dose of 800 mg Q4W (two injections) was optimized based on the safety, PK, and PD profiles observed in patients in the previous group.

[0124] The planned SAR113244 treatment regimen was as follows:

[0125]

Table 1

[0126] In some cases, group 3 was not conducted (the highest dose administered was 500 mg), and group 2 was terminated early because the preliminary study results indicated that 500 mg would be an appropriate dose level for the next study.

[0127] Selection and timing of administration for each patient Patients were assigned to treatment according to a randomization list.

[0128] SAR113244 or placebo was initiated in the first patient at approximately 8:00 AM and administered to patients in a fasting state on the morning of day 1 and the morning of day 29. After administration, the patients remained at the trial facility until the morning of the next day (i.e., day 2 and day 30) and were discharged from the facility after the evaluations on the morning of day 2 and day 30.

[0129] Previous and concomitant treatments The use of concomitant medications was prohibited during the trial unless specified in the eligibility criteria or trial procedures or medically necessary. However, if specific treatments were required for any reason, accurate records including the name of the drug (international nonproprietary name), daily dose, and duration of use were entered into the eCRF.

[0130] Pharmacodynamics, Safety, and Pharmacokinetics Assessment The overview of safety, PK, and PD assessments for the test procedures was shown in the test flow chart (Table 2); the detailed schedule for the treatment period was shown in the period flow chart (Table 3).

[0131]

Table 2

[0132]

Table 3

[0133]

Table 4

[0134]

Table 5

[0135]

Table 6

[0136]

Table 7

[0137]

Table 8

[0138]

Table 9

[0139] Pharmacodynamics Assessment The detailed schedule for PD assessment was shown in Table 2.

[0140] CXCR5 Receptor Occupancy CXCR5 receptor occupancy (RO) by SAR113244 on the cell surface of B lymphocytes (total B lymphocyte target population [CD19+]), naive sub-target population (CD19+ / IgD+ / CD27-), and memory sub-target population (CD19+ / CD27+; or CD19+ / IgD- / CD27-) in whole blood samples was evaluated using a valid assay. This assay was established using quantitative flow cytometry and the use of calibration beads (CellQuant Calibrator kit) to convert the fluorescence intensity of the sample to a value corresponding to the number of antibodies bound per cell. The lower limit of quantification (LLOQ) of the assay was 20%.

[0141] CXCL13 CXCL13 was quantified in human serum using a valid enzyme-linked immunosorbent assay (ELISA) method. Considering the minimum required dilution of the assay, the LLOQ and upper limit of quantification were 15.6 and 500 pg / mL, respectively. CXCL13 was also quantified in urine using a diagnostic ELISA method.

[0142] Analysis of Biomarkers of B Cell Subsets and T Cells Analysis of B and T cell subsets was performed using flow cytometry. In addition to the identification of cells by their size (forward scatter) and granulation (side scatter), several cell subpopulations were identified by the expression of their specific cell surface markers.

[0143] Using the characteristics of B and T cell subtypes (e.g., naive cells, memory cells, antibody-secreting cells [i.e., plasmablasts, plasma cells]) based on these markers, the biological effects and potential of the biomarkers under SAR113244 treatment were investigated. Disease-Related Parameters · SELENA-SLEDAI score. · British Isles Lupus Assessment Group (BILAG) score. · Anti-Smith, Anti-Ro, Anti-La, Anti-cardiolipin (IgG, IgM). · Anti-dsDNA antibody and ANA levels. · Plasma complement levels (C3, C4). · Physician Global Assessment (PGA), Lupus-quality of life (QoL) and Functional Assessment of Chronic Illness Therapy (FACIT)-Fatigue. · Blood sedimentation rate and C-reactive protein (CRP).

[0144] Safety variables and timing of assessments Patients were monitored for safety by adverse events (AEs) spontaneously reported by the patients or observed by the study physicians, clinical laboratory evaluations (hematology, biochemistry, and urinalysis), serum Ig, peripheral blood B and T cells, anti-SAR113244 antibody, vital sign measurements, 12-lead electrocardiogram (ECG), ECG morphology, physical examinations, body weight, body temperature, and local tolerability in injection site assessments.

[0145] The QuantiFERON TB Gold test was performed only at screening. Serology (HBsAg, hepatitis B virus core antibody, hepatitis C virus antibody, anti-HIV1 and anti-HIV2 antibodies, Epstein-Barr virus, CMV, positive IgM antibody titer in the presence of negative IgG titer against hepatitis A virus) was performed at screening as well; urine drug screening and urine alcohol tests were performed at screening and on Day 1. For women, a serum pregnancy test was performed at screening, followed by randomization and a urine pregnancy test.

[0146] The schedule of safety assessments is shown in Tables 2 and 3.

[0147] Adverse events From the ICF signs until the patient's visit to the EOS, all AEs were recorded regardless of their severity or relationship to the IMP. The principal investigator of the clinical trial described them in detail in the eCRF, including their relationship to the IMP.

[0148] A serious adverse event (SAE) is any untoward medical occurrence at any dose: · resulting in death, or · threatening life, or · requiring hospitalization or prolonging existing hospitalization, or · causing persistent or significant disability / incapacity, or · congenital anomaly / birth defect, or · medically important event.

[0149] Safety parameters of clinical examinations Standard clinical examination parameters (biochemistry, hematology, and urinalysis) were measured.

[0150] Other safety parameters 1. Vital signs Heart rate, systolic blood pressure (SBP), and diastolic blood pressure (DBP) were measured 10 minutes after lying quietly in the supine position and 3 minutes after standing.

[0151] 2. Electrocardiogram A 12-lead ECG was recorded at least 10 minutes after lying in the supine position (recorded for 10 seconds at 25 mm / s, 10 mm / mV).

[0152] The ECG profile was evaluated by 12-lead ECG and was evaluated several times by testing at the ECG Core Lab (semi-automatic reading) and by reading at the center at all time points except screening. Each time point was a single recording.

[0153] The following parameters were received from the ECG reading center. · RR (ms), the average from all sinus rhythm complexes recorded over 10 seconds. ·From the average RR of all sinus rhythm complexes recorded over 10 seconds of HR (bpm). ·PR (ms), overall interval measurement from the superposition of the individual average heartbeats of 12 leads. ·QRS (ms), overall interval measurement from the superposition of the individual average heartbeats of 12 leads. ·QT (ms), overall interval measurement from the superposition of the individual average heartbeats of 12 leads. ·QT interval corrected for heart rate using Bazett's formula (QTcB) (ms), Bazett's correction (QTcB = QT.RR - 0.50) from the overall QT and average RR. ·QT interval corrected for heart rate using Fridericia's formula (QTcF) (ms), Fridericia's correction (QTcF = QT.RR - 0.33) from the overall QT and average RR.

[0154] Morphological analysis of the ECG waveform was also performed.

[0155] 3. Local tolerance at the injection site The skin around the SC injection area was examined for its potential reaction to SC injection (reaction was observed at a predetermined time or any time). Findings were described separately for each injection and for each injection site (when administration of each dose was carried out by multiple injections at the same time point). The maximum diameters of erythema and swelling (including induration and / or edema) were measured and recorded separately in millimeters. Erythema and swelling were stratified separately. A value of 0 was recorded if there was no change in the parameters expressed by treatment at the time of observation.

[0156] Furthermore, any injection site reaction of "severe" or worse intensity, or all injection site reactions lasting longer than 24 hours regardless of intensity, were reported as particularly notable AEs (AESIs).

[0157] The following symptoms or presence or absence of single observations were recorded without stratification: erosion, dryness, scaling, cracking, crusting, and glazing. The decision as to whether any of each unstratified tolerability observations should be reported as an AE was the responsibility of the principal investigator of the clinical trial.

[0158] 4. Immunoglobulins Blood samples for serum immunoglobulins (IgG, IgM, IgD, IgE, IgA) were taken at the time points shown in Tables 2 and 3.

[0159] 5. Total peripheral blood B and T cells Blood samples for total peripheral blood B and T cells were taken at the time points shown in Tables 2 and 3.

[0160] Pharmacokinetics assessment and timing Pharmacokinetics measurement and timing The pharmacokinetics sampling times are shown in the study flow chart in Table 2 and the period flow chart in Table 3.

[0161] The concentration of SAR113244 in plasma was determined by a validated ELISA technique at the LLOQ of 0.040 μg / mL.

[0162] Furthermore, the sampling times for immunogenicity assessment are shown in the study flow chart (Table 2). The ADA assay was performed using a validated ELISA method. All samples were first evaluated using a screening assay. Samples found positive in the screening assay were then tested in a confirmatory assay. Titers were reported only for samples confirmed to be positive.

[0163] Pharmacokinetic variables The plasma concentration of SAR113244 and relative real-time values were used to calculate the PK parameters listed in Table 4 using a non-compartmental method by validated software (WinNonlin version 5.2.1, PKDMS version 2.2 run by Certara).

[0164]

Table 10

[0165] Validity of Measurement Standard measurements are appropriate for the analysis of the safety and tolerability, PD, and PK of SAR113244 used in this study.

[0166] Statistical Considerations Determination of Sample Size The sample size for this study was based on empirical considerations. The calculation of the sample size was not performed.

[0167] Population for Analysis Population for Pharmacodynamics All patients with sufficient and explainable main PD data and without major or important protocol deviations were included in the PD population. Patients treated with placebo were not included in the analysis of RO data.

[0168] Population for Safety All patients exposed to the study treatment, regardless of the amount of treatment administered, were included in the safety population.

[0169] Population for Pharmacokinetics All patients with sufficient and explainable main PK data and without major or important deviations regarding the uptake of the test drug were included in the PK population. Patients receiving only placebo were not included in the PK population.

[0170] Statistical Analysis Demographics and Baseline Characteristics Demographic variables were summarized by treatment group and overall using descriptive statistics in the safety population. Continuous data were summarized using available data, mean, standard deviation (SD), median, minimum, and maximum numbers. Categorical and ordinal data were summarized using the number and percentage of patients in each treatment group. All data were enumerated.

[0171] The medical history or surgical history was coded using the Medical Dictionary for Regulatory Activities (MedDRA, version 18.1). All reported medical history or surgical history was presented by major organ system classification (SOC) and high-level terms, by treatment group and overall. All medical history data, including any family history of autoimmune disease in any close relative, was listed.

[0172] Baseline disease characteristics were summarized by treatment group and overall using descriptive statistics in the safety population. Continuous data were summarized using available data, mean, SD, median, minimum and maximum, and the number of first and third quartiles; the number of attributable values was also provided for anti-dsDNA antibody levels. Categorical and ordinal data were summarized using the number and percentage of patients in each treatment group.

[0173] Pre- or concomitant medication The following medications were reported on the appropriate eCRF pages: · Pre- and concomitant corticosteroids were taken for lupus within 5 years before study entry and throughout the entire study. · Pre- and concomitant medications other than corticosteroids were taken for lupus within 6 months before study entry and throughout the entire study. · Pre- and concomitant medications were not associated with lupus within 3 months before study entry and throughout the entire study.

[0174] All medications were coded using the World Health Organization - Drug Dictionary, 2015 SEP version.

[0175] Medications were summarized by treatment group as described in the World Health Organization - Drug Dictionary. Pre- and concomitant medications were summarized for the safety population. All medications were listed.

[0176] Degree of drug exposure and medication compliance in the clinical trial Treatment exposure (i.e., number of days of administration) was summarized by treatment group in the safety population.

[0177] Details of drug administration (actual treatment received, dates and times of IMP administration, intended and actual doses received, patients receiving IMP from a specific batch, and randomization scheme) were listed.

[0178] Analysis of pharmacodynamic endpoints All PD analyses were performed using the PD population.

[0179] 1. Disease-related markers The following markers were analyzed as raw data, absolute and percentage changes from baseline, unless otherwise specified. Only data evaluated by Covance at the center was considered. · Anti-dsDNA antibodies. · ANA levels: negative / positive and titer if positive. · Plasma complement levels C3 and C4. · Blood sedimentation rate and CRP. · Anti-Smith, anti-Ro, anti-La, anti-cardiolipin (IgG, IgM): negative / positive. · Anti-Smith, anti-Ro, anti-La, anti-cardiolipin (IgG, IgM): negative / positive. 2. Disease activity and quality of life scales · SELENA-SLEDAI scores analyzed as raw data and absolute changes from baseline. · BILAG scores (A, B, C, D, E) at each visit for each organ-based system. · PGA scores: values on a visual analog scale in mm, analyzed as raw data and absolute changes from baseline. · Lupus-QoL total scores in the range from 0 to 100 (see 16-1-9-sap [Annex F]) were analyzed as raw data and absolute changes from baseline. · FACIT-Fatigue total scores (see 16-1-9-sap [Annex G]) were analyzed as raw data and absolute changes from baseline.

[0180] 3. CXCR5 receptor occupancy on peripheral B cells The following parameters of CXCR5 RO(%) were derived as follows: · CXCR5 normalized RO%. · The period of saturation of CXCR5 by SAR113244 in the days following the first dose, where saturation was defined as a normalized RO > 80% (based on the accuracy of the assay method: ±20%).

[0181] 4. Serum CXCL13 levels Serum CXCL13 levels were analyzed using raw data, absolute and percentage changes from baseline, where the baseline value was the pre-dose value on Day 1.

[0182] 5. Peripheral blood B and T cell subsets Blood B and T cell subsets were analyzed as raw data, absolute and percentage changes from baseline and were performed at the center using flow cytometry.

[0183] 6. Analysis All analyses were descriptive only. No formal statistical tests were performed.

[0184] Descriptive statistics were provided by treatment group using available data, mean, SD, standard error of the mean (SEM), median, minimum and maximum, and the number of the 1st and 3rd quartiles; the number of attributable values was also provided where applicable. Categorical and ordinal data were summarized using the number and percentage of patients in each treatment group.

[0185] Mean time profile plots (±SEM) of raw data, absolute and / or percentage changes from baseline (by parameter) were also generated by treatment group for the selected parameters.

[0186] Receptor occupancy data: · RO and standardized RO data were summarized by treatment group and visit using descriptive statistics (mean, median, minimum, and maximum) and plots; individual plots were generated if needed. · The duration of receptor saturation was described using the median, minimum, and maximum by treatment group.

[0187] Selected individual data were listed.

[0188] Analysis of safety data Safety assessments were based on individual values (clinically significant abnormalities) and summaries of descriptive statistics.

[0189] All safety analyses were conducted using the safety population.

[0190] 1. Adverse events Adverse events were coded according to MedDRA (version 18.1).

[0191] Adverse events were classified into predefined standard categories according to the following criteria: · Pre-treatment AE: AE that occurred during the pre-treatment phase. · AE occurring during treatment (TEAE): AE that occurred during the treatment phase. · Post-treatment AE: AE that occurred during the post-treatment phase.

[0192] All AEs reported during the trial were listed, and if any, the comments recorded in the eCRF related to the AE were listed.

[0193] AEs occurring during treatment were assigned to the treatment received prior to the time of AE onset.

[0194] The following frequency distributions of TEAE (incidence tables) were provided for the safety population: · Summary of TEAE: Number and percentage of patients with TEAE, severe TEAE, serious TEAE, TEAE resulting in death, TEAE resulting in discontinuation of maintenance treatment, and AESI occurring during treatment. · Summary of TEAE by main SOC and preferred terms (PT): · Number and percentage of patients with at least one TEAE · Number and percentage of patients and events.

[0195] All TEAEs, whether related or unrelated to the IMP, were summarized by SOC.

[0196] Any deaths, SAEs, AEs, or AESIs that led to treatment discontinuation were listed.

[0197] 2. Clinical laboratory evaluations All individual data related to planned hematology and biochemistry, including reconfirmed values, were listed by biological function, patient, and visit. If available, data from unscheduled clinical tests were also listed. In these listings, individual data were flagged if they were below or above the lower or higher assay limits, and / or reached the absolute limits of abnormal (PCSA) criteria that should be considered clinically significant.

[0198] The value used as the baseline was the T0H (pre-dose) assessment value on Day 1. If any of the scheduled baseline tests were repeated in any patient, the last reconfirmed value was considered the baseline if it was performed prior to the first pre-dose of IMP administration.

[0199] For parameters by test range and / or abnormal criteria (PCSA), all post-baseline assessments performed during the treatment phase, including reconfirmed values, were used for in-treatment analysis. The number of patients with in-treatment abnormalities (PCSA) was stratified by baseline status (normal, abnormal) and presented by treatment group. This analysis was also performed for abnormal test range values.

[0200] All individual data regarding planned hematology and biochemistry, including reconfirmed values, were enumerated by biological function. If any, data from unscheduled clinical tests were also enumerated. In these enumerations, individual data were flagged if they were below or above the lower or higher assay limits, respectively, and / or reached the absolute limits of the PCSA criteria, as defined.

[0201] An enumeration of individual post-baseline abnormalities by patient was provided.

[0202] An enumeration related to an increase in ALT ≥ 2 × ULN was also provided.

[0203] An enumeration of patients with combined PCSA of liver function was also provided. Reported were patients related to all time points (planned and reconfirmed) of the study.

[0204] All qualitative and quantitative urine test results (dipstick), including reconfirmed values, were enumerated.

[0205] 3. Vital Signs Heart rate, SBP and DBP, and body weight were analyzed as raw parameter values and changes from baseline.

[0206] Oral temperature was analyzed as raw data.

[0207] The values used for baseline were the T0H (pre-dose) values on Day 1, except for body weight where the baseline was the value on Day 1. If any of the planned baseline tests were repeated in any patient and were performed before IMP administration, the reconfirmed values were considered as the baseline.

[0208] For all parameters, on-treatment analysis was performed using all post-baseline assessments during the treatment phase, including reconfirmed values. The number of patients with on-treatment abnormalities (PCSA) was provided regardless of the normal or abnormal status at baseline and presented by treatment.

[0209] For heart rate, SBP, and DBP, changes from raw data and baseline were summarized with descriptive statistics for each type of measurement, parameter, and time point by treatment.

[0210] For weight, percentage changes from raw data and baseline were provided by treatment and scheduled time.

[0211] For oral temperature, summaries of raw data were provided by treatment and scheduled time.

[0212] All individual data, including rechecked values, were listed. During listing, values were flagged if they reached the limits of the PCSA criteria or were defined.

[0213] Separate listings of individual data from patients with PCSA after baseline were provided.

[0214] 4. Electrocardiogram The following parameters: HR, PR, QRS, QT, QTcB, and QTcF were received from the ECG reading center and analyzed.

[0215] The value used for baseline was the T0H (pre-dose) assessment value on Day 1. If any of the scheduled baseline tests were repeated in any patient and were performed before IMP administration, the last rechecked value was considered the baseline.

[0216] All parameters were analyzed as absolute changes from raw data and baseline. Additionally, for PCSA analysis, PR, and QRS, percentage changes from baseline were also analyzed.

[0217] For all parameters, "during treatment" analyses were performed in the safety population using all assessments during the treatment phase, including any unplanned / reconfirmed values. The number of patients with PCSA was provided in the summary table regardless of baseline normal or abnormal status. This table was presented by treatment.

[0218] Descriptive statistics (raw data and absolute changes from baseline) were provided by visit and treatment group.

[0219] Patients by morphological assessment were summarized by High Level comments and comments selected by comment (ECG Core Lab code list).

[0220] The following listings were generated: · Individual raw data and absolute changes from baseline for HR, PR, QRS, QT, QTcB, QTcF, · Individual data for any patients with PCSA after baseline, · Patients with QTcB / QTcF > 480 ms and / or changes from baseline of QTcB / QTcF > 60 ms, · Patients with at least one abnormality in quantitative assessment (i.e., abnormal 12-lead ECG) after the first dose, and · All morphological comments.

[0221] 5. Other related safety parameters a. Anti-SAR113244 antibody The ADA analysis was based on all randomized and treated (positive, negative or indeterminate) patients with at least one evaluable post-baseline ADA sample. The ADA analysis was based on baseline (before dose on Day 1) and all post-baseline ADA assessments during the treatment phase and follow-up period.

[0222] The following descriptive statistics were provided by treatment group and overall population: · ADA status at baseline: · The number and percentage of patients with positive, negative, or indeterminate ADA samples at baseline. · Descriptive statistics of baseline titers for patients with pre-existing ADA. · ADA status during the baseline period: · The number and percentage of patients with evaluable, positive, negative, or indeterminate ADA samples. · For patients with positive ADA: · Descriptive statistics of peak titers for patients with positive ADA, · For patients with negative ADA at baseline: · n(%) of patients with treatment-induced ADA, · Descriptive statistics of peak titers for patients with treatment-induced ADA. · For patients with pre-existing ADA: · Descriptive statistics of peak titers, · n(%) of patients with treatment-boosted ADA, · Descriptive statistics of peak titers for patients with treatment-boosted ADA, · Ratio of titers from baseline to post-baseline, · X-fold increase in titers calculated as the ratio of post-treatment titers to baseline titers for patients with treatment-boosted ADA. · ADA prevalence, · ADA incidence.

[0223] b. Local tolerability at the injection site Descriptive statistics of the highest grade of presence of all or at least one reaction during the entire trial were summarized by treatment group.

[0224] All data were enumerated.

[0225] c. Serum immunoglobulins Serum IgA, IgD, IgM, IgE, and IgG were evaluated.

[0226] Raw data, absolute and percentage changes from baseline were summarized using descriptive statistics by treatment and scheduled time points. The value used as baseline was the Day 1 pre-dose assessment.

[0227] All data were enumerated.

[0228] d. All peripheral blood B and T cells Raw data, absolute and percentage changes from baseline were summarized using descriptive statistics by treatment and scheduled time points. The value used as baseline was the Day 1 pre-dose assessment.

[0229] All data were enumerated.

[0230] Due to operational issues in cohort 1, all peripheral blood B and T cells were not measured during the study. Only screening values were available for cohort 1.

[0231] Pharmacokinetic data analysis The plasma PK parameters and concentrations of SAR113244 were summarized by descriptive statistics (mean, geometric mean, median, SD, SEM, coefficient of variation (CV), minimum, and maximum) for each treatment. ·C max and AUC 0~4w For, accumulation was evaluated by a linear fixed-effect model using the dose as a fixed effect in SAS PROC MIXED, Log(ratio of Day 29 to Day 1) = dose + error as evaluated by.

[0232] The accumulation ratio of Day 29 to Day 1 was evaluated separately at each dose level and pooled across dose levels within the fixed-effect model framework. The accumulation ratio was evaluated by estimating the ratio of Day 29 to Day 1 on the log scale corresponding to the 90% confidence interval (CI), and further transformed to the Day 29 / Day 1 accumulation ratio corresponding to those 90% CIs using back-transformation.

[0233] An enumeration of the individual accumulation ratios was provided along with their descriptive statistics. ·t 1 / 2z For t, the dose effect was evaluated on Day 29 by a linear fixed effect model. Log(t 1 / 2z ) = dose + error

[0234] t 1 / 2z Point estimates and 90% CIs of the geometric mean of t were provided and pooled separately for each dosing group across dose levels.

[0235] t max The distribution of t values was represented by a histogram plot for each dose level and day. ·C max and AUC 0~4w The dose proportionality of C and AUC was evaluated using pairwise comparisons of the log-transformed parameters. Point estimates and 90% CIs were provided. ·Dose, day, and day * Dose interactions were modeled as random effects, fixed effects, and patient (dose) in the following linear mixed effect model framework: Log(parameter) = dose + patient(dose) + day + day * dose + error By equating the observed and predicted mean squares within and overall SD of log(C max ) and log(AUC 0~4w ) were estimated.

[0236] Interleukin analysis Interleukin analysis was not performed.

[0237] Changes to the conduct of the trial or planned analyses Changes to the conduct of the trial There were four amendments to the protocol. The changes explain the conduct of the trial as presented in this report. A summary of the changes included in the amendments is provided in Table 5.

[0238]

Table 11

[0239] Change in planned analysis From protocol to statistical analysis plan The following major modifications to the statistical section of the protocol were as follows: · Detailed PD analysis was included. · Analysis of ECG data was updated to take into account reading by the data center and was updated. · Analysis of the anti - SAR113244 antibody was updated according to the latest Sanofi analysis guidelines. · Enumeration of some individual data was generated as needed.

[0240] After database lock Maximum receptor binding (normalized and non - normalized maximum RO%) and time to first maximum achieved (normalized and non - normalized tmax[RO%]) were not calculated as they were considered not relevant.

[0241] The PK parameter tlast was not calculated.

Example

[0242] Test patients Patient allocation As planned, 8 patients were enrolled in cohort 1. Although 16 patients were planned to be enrolled in cohort 2, enrollment was difficult, so cohort 2 ended early after enrollment of 13 patients. Cohort 3 (administering an 800 mg dose) was not carried out due to prior indications that 500 mg was an appropriate dose level for the advancement of SAR113244. These decisions were not related to any concerns regarding the safety or tolerability of SAR113244.

[0243] Of the 21 lupus patients randomized and treated in this trial, 19 completed the treatment period of the trial. A total of 16 patients received SAR113244 and 5 patients received placebo (Table 6).

[0244]

Table 12

[0245] One patient receiving 250 mg of SAR113244 Q4W and one patient receiving 500 mg of SAR113244 Q4W discontinued their consent and prematurely interrupted the trial.

[0246] Randomization and dosing irregularities A total of two patients had minor protocol deviations related to dosing irregularities.

[0247] Patient number 276001024 and patient number 276001041 had already been dosed with 500 mg of SAR113244 Q4W before the trial responsible physician contacted the IWRS. However, it was confirmed on site by the unblinded CRA that corrective measures had been given as later assigned by the IWRS (i.e., no randomization error occurred).

[0248] Analyzed dataset All 21 patients were included in the safety and PD populations, and all patients receiving SAR113244 were included in the PK population (Table 7).

[0249]

Table 13

[0250] Demographics and other baseline characteristics Demographics Demographic characteristics at baseline are shown in Table 8.

[0251]

Table 14

[0252] At baseline, 7 / 10 (70.0%) of the patients receiving 500 mg Q4W SAR113244 were <45 years of age, compared to 1 / 5 (20.0%) of the patients receiving placebo. Three (50.0%) of the six patients receiving 250 mg Q4W SAR113244 were <45 years of age.

[0253] The majority (19 / 21) of the patients were female. All of the patients receiving SAR113244 were female, and 3 / 5 (60.0%) of the patients receiving placebo were female. The majority (20 / 21) of the patients were Caucasian / White, and 1 / 21 of the patients receiving 500 mg Q4W SAR113244 were Asian / East Asian.

[0254] Medical and Surgical History The most frequently reported medical or surgical history other than SLE was vascular hypertension disorder, reported in eight patients across the treatment groups. Age-related problems were reported in five patients across the treatment groups. Depressive disorders, joint treatment procedures, and hypothyroid disorders were reported in four patients each across the treatment groups, and fat-soluble vitamin deficiencies and disorders were reported in three patients across the treatment groups. All other medical or surgical histories other than SLE were reported in one or two patients across the treatment groups.

[0255] Disease Characteristics at Baseline A summary of the disease characteristics is shown in Table 9. A summary of the BILAG scores at baseline is shown in Table 10.

[0256]

Table 15

[0257]

Table 16

[0258]

Table 17

[0259]

Table 18

[0260] Overall, the mean disease duration was approximately 10 years. The mean disease durations for patients receiving placebo and 500 mg of SAR113244 were approximately 9 and 8 years, respectively, and the mean disease duration for patients receiving 250 mg of SAR113244 was approximately 15 years.

[0261] All patients who were tested positive for ANA with a titer ≥ 1:160 at screening (measured by each laboratory [Synlab]; these values were used appropriately). One patient (in the placebo treatment group) was tested negative for ANA at baseline, 6 / 20 patients were tested positive for ANA with a titer ≤ 1:80, and 14 / 20 patients were tested positive for ANA with a titer ≥ 1:160 at baseline. Baseline values were measured by C ovance.

[0262] Overall, 18 / 21 patients had a positive titer for anti-dsDNA antibodies at screening (measured by Charite CRO) and 6 / 21 patients had a positive titer for anti-dsDNA antibodies at baseline (measured by Covance).

[0263] The overall total SELENA - SLEDAI score ranged from 2 to 8 at screening and from 4 to 10 at baseline. The overall PGA of disease activity ranged from 9 to 52 mm on day 1 (100 mm indicates severe disease activity). Overall, 12 / 21 (57.1%) patients received ongoing treatment with prednisone (total dose ranging from 2.5 to 15 mg) at screening.

[0264] All patients had negative QuantiFERON TB Gold test results at screening.

[0265] Pre - and / or concomitant medications taken for lupus Two out of five (40.0%) patients receiving placebo, four out of six (66.7%) patients receiving 250 mg Q4W SAR113244, and seven out of ten (70.0%) patients receiving 500 mg Q4W SAR113244 were treated for lupus with corticosteroids during the trial.

[0266] Four out of five (80.0%) patients receiving placebo, four out of six (66.7%) patients receiving 250 mg Q4W SAR113244, and nine out of ten (90.0%) patients receiving 500 mg Q4W SAR113244 were treated for lupus with concomitant medications other than corticosteroids during the trial. The most frequent of these concomitant medications were antiprotozoal agents, anti - inflammatory and antirheumatic products, and immunosuppressants.

[0267] Pre - and / or concomitant medications not related to lupus Prior to the first administration of IMP, no patients were vaccinated with medications not related to lupus.

[0268] Five out of five (100.0%) patients receiving placebo, five out of six (83.3%) patients receiving 250 mg Q4W SAR113244, and eight out of ten (80.0%) patients receiving 500 mg Q4W SAR113244 were treated with concomitant medications not related to lupus during the trial. The most frequent of these concomitant medications were vitamins, drugs for acid - related disorders, analgesics, and agents acting on the renin - angiotensin system.

[0269] Measurement of treatment compliance All patients except two who withdrew from the trial before day 29 received two IMP injections as planned.

Example

[0270] Pharmacodynamic evaluation Pharmacodynamic endpoints CXCR5 receptor occupancy on B cells The mean SAR113244 occupancy of CXCR5 on peripheral blood B cells over time is shown in Figure 2.

[0271] Individual RO values and their descriptive statistics are shown in Tables 32 to 33. Patients treated with placebo were not included in the analysis of receptor occupancy data. Data from 6 patients in the 250 mg dose group and 9 patients in the 500 mg dose group (patient number 276001019 was excluded) were included. A summary of these statistics is shown in Table 11.

[0272] CXCR5 occupancy was quantified after a single 250 mg and 500 mg SC dose of SAR113244, and the corresponding values reached steady state from 7 days after dosing in all patients (day 8 is the first sample collection time after dosing). 84 days after dosing (i.e., 56 days after the second dose), in some patients in both groups, CXCR5 decreased to <LLOQ drug levels and a decrease in CXCR5 occupancy was observed (patient numbers 276001006, 276001014, and 276001024), but most patients still had CXCR5 occupied by the drug on day 113 (EOS) as shown below. · RO% in the range between 53.7% and 78.9% for 2 out of 5 patients at 250 mg. · RO% in the range between 29.6% and 83.4% for 7 out of 9 patients at 500 mg.

[0273] In the RO assay, it was found that the maximum RO varied between patients. Therefore, to standardize the maximum RO across patients, individual RO results were standardized based on a normalization factor determined by adding a pre-dose sample saturating the concentration of SAR113244 for each patient (to determine the maximum RO achievable by each patient in the assay). The mean normalized SAR113244 binding to the CXCR5 receptor on peripheral blood B cells over time is shown in Figure 3. The "saturation period" of the normalized RO% was defined as the time interval during which the occupancy of CXCR5 by SAR113244 was maximal. Based on the accuracy of the assay method (i.e., ±20%), maximum saturation of CXCR5 was reached when RO > 80%.

[0274] The individual standardized RO values and their descriptive statistics are shown in Tables 34 to 35. A summary of these statistics is shown in Table 12.

[0275] The individual values of the maximum occupancy period and their descriptive statistics are shown in Table 36. A summary of these descriptive statistics is shown in Table 13.

[0276]

Table 19

[0277]

Table 20

[0278]

Table 21

[0279] Saturation of CXCR5 by SAR113244 occurred in all patients at 250 mg and 500 mg by 7 days after the first dose (first RO sample collection time after dosing). Over the course of the study, the period of saturation associated with the second dose appeared to increase to a median of 42 days at 250 mg and 56 days at 500 mg.

[0280] CXCR5 occupancy was <LLOQ for the standardized RO% in two patients (patient numbers 276001006 and 276001024) and began to decline at 84 days after the first dose in several patients. On day 112 after the first dose, detectable CXCR5 RO by SAR113244 was observed in the same patient, but the normalized RO% continued to decrease: the normalized RO% ranged between 34.1% and 75.5% mainly in six patients at the 500 mg dose level, and saturation of CXCR5 occupancy was observed at normalized RO% of 89.0% and 85.8% in patient numbers 276001011 and 276001041, respectively. The distribution of patients by different normalized RO% ranges is shown in Table 14.

[0281]

Table 22

[0282] Disease-related markers Anti-dsDNA antibody One patient (patient number 276001003) in the 250 mg SAR113244 treatment group and one patient (patient number 276001024) in the 500 mg SAR113244 treatment group had abnormally high values for anti-dsDNA antibody at baseline. The values remained above the ULN throughout the study for both patients.

[0283] There was little change in the mean percentage change from baseline in anti-dsDNA antibody values across treatment groups at each scheduled visit.

[0284] ANA level In the placebo treatment group, 4 / 5 (80%) of the patients were tested positive for ANA on day 1 (2 / 5 at a titer of 1:640, 1 / 5 at a titer of 1:160, 1 / 5 at a titer of 1:320), and 5 / 5 (100%) of the patients were tested positive on day 113 (1 / 5 at a titer of 1:40, 1 / 5 at a titer of 1:160, 1 / 5 at a titer of 1:320, 1 / 5 at a titer of 1:640, 1 / 5 at a titer of 1:2560).

[0285] All patients in the 250 mg and 500 mg SAR113244 treatment groups with analyzable samples were tested positive for ANA on both Day 1 and Day 113.

[0286] In the 250 mg SAR113244 treatment group, on Day 1, 3 / 6 patients had a titer of 1:80 and 3 / 6 patients had a titer of 1:160. On Day 113, one patient's sample was lost, 1 / 5 patients had a titer of 1:40, 1 / 5 patients had a titer of 1:80 , 1 / 5 patients had a titer of 1:160, and 2 / 5 patients had a titer of 1:320.

[0287] In the 500 mg SAR113244 treatment group, on Day 1, 2 / 10 patients had a titer of 1:40, 1 / 10 patients had a titer of 1:80, 4 / 10 patients had a titer of 1:160, 2 / 10 patients had a titer of 1:320, and 1 / 10 patients had a titer of 1:640. On Day 113, 1 / 10 patients had a titer of 1:40, 5 / 10 patients had a titer of 1:160, 2 / 10 patients had a titer of 1:320, 1 / 10 patients had a titer of 1:640, and 1 / 10 patients had a titer of 1:2560.

[0288] Plasma complement levels C3 and C4 One of the five patients in the placebo treatment group had abnormal baseline plasma complement level C3. Four of the six patients in the 250 mg SAR113244 treatment group had abnormal baseline plasma complement level C3, and 3 / 6 patients had abnormal baseline plasma complement level C4. Five of the nine patients in the 500 mg SAR113244 treatment group had abnormal baseline plasma complement level C3, and 3 / 9 patients had abnormal baseline plasma complement level C4.

[0289] No mean percentage change from baseline in plasma complement level C3 or C4 or any treatment - or administration - related trends in the normalization of C3 or C4 values were seen at each scheduled visit.

[0290] Blood sedimentation rate (SED rate) and C-reactive protein (CRP) Of the five patients in the placebo treatment group, one and one-sixth of the patients receiving 250 mg of SAR113244 had abnormal blood SED rates at baseline. Of the ten patients in the 500 mg SAR113244 treatment group, two had abnormal blood SED rates at baseline and one-tenth had abnormal CRP values at baseline.

[0291] No treatment- or administration-related trends were observed in the mean percentage change from baseline in blood SED rate or CRP from baseline to each scheduled visit.

[0292] Anti-Smith, anti-Ro / SS-A, anti-La / SS-B, anti-cardiolipin antibodies Not all patients had analyzable samples for assessment of anti-Smith, anti-Ro, anti-La, and anti-cardiolipin antibodies.

[0293] All patients with analyzable samples in the placebo treatment group and the 500 mg SAR113244 treatment group were tested negative for anti-Smith antibodies on day 1 and day 113. Four out of five patients (80.0%) in the 250 mg SAR113244 treatment group were tested negative for anti-Smith antibodies on day 1, and all patients in the 250 mg SAR113244 treatment group with analyzable samples were tested negative for Smith antibodies on day 113. On day 1 and day 113, 3 / 5 (60.0%) and 2 / 4 (50.0%) of the patients in the placebo treatment group were tested negative for anti-Ro / SS-A antibodies, respectively. On day 1 and day 113, 4 / 5 (80.0%) of the patients in the 250 mg SAR113244 treatment group were tested negative for anti-Ro / SS-A antibodies. On day 1 and day 113, 7 / 8 (87.5%) and 8 / 10 (80.0%) of the patients in the 500 mg SAR113244 treatment group were tested negative for anti-Ro / SS-A antibodies, respectively.

[0294] On Day 1 and Day 113, 4 / 5 (80.0%) and 3 / 4 (75.0%) of the patients in the placebo treatment group, respectively, were tested negative for anti-La / SS-B antibodies. All patients with analyzable samples in the 250 mg SAR113244 treatment group were tested negative for anti-La / SS-B antibodies on both Day 1 and Day 113. On Day 1 and Day 113, 7 / 8 (87.5%) and 9 / 10 (90.0%) of the patients in the 500 mg SAR113244 treatment group, respectively, were tested negative for anti-La / SS-B antibodies.

[0295] All patients with analyzable samples in the placebo and 250 mg SAR 113244 treatment groups were tested negative for anti-cardiolipin antibodies (IgG) on Day 1 and Day 113. On Day 1, 8 / 9 (88.9%) of the patients in the 500 mg SAR113244 treatment group were tested negative for anti-cardiolipin antibodies (IgG), and 1 / 9 (11.1%) of the patients were tested positive. On Day 113, 9 / 10 (90.0%) of the patients in the 500 mg SAR113244 treatment group were tested negative for anti-cardiolipin antibodies (IgG), and 1 / 10 (10.0%) of the patients were tested positive.

[0296] All patients with analyzable samples across treatment groups were tested negative for anti-cardiolipin antibodies (IgM) on Day 1 and Day 113.

[0297] Disease Activity and Quality of Life Scales The mean total SELENA-SLEDAI score was similar across treatment groups throughout the study and ranged from 0 to 10. The mean change from baseline in the total score was <1.3 throughout the study for all treatment groups.

[0298] The BILAG scores at baseline compared to Day 113 were similar. SAR113244 did not appear to have an effect on the BILAG scores.

[0299] The mean PGA score was similar across the treatment groups up to day 113 from baseline. In the 500 mg SAR113244 treatment group, the mean change from baseline in the PGA score on day 113 was an increase of 8.8, compared with a decrease of 5.6 and an increase of 0.5 in the placebo and 250 mg SAR113244 treatment groups, respectively.

[0300] The Lupus-QoL total score at baseline was similar compared to day 113. SAR113244 did not appear to have an effect on the Lupus-QoL score.

[0301] The FACIT-Fatigue total score at baseline was similar compared to day 113. SAR113244 did not appear to have an effect on the FACIT-Fatigue score.

[0302] Serum CXCL13 levels Treatment- or administration-related trends were not evident in the CXCL13 data from baseline time to day 113.

[0303] Peripheral blood B and T cell subsets B cell subsets The frequencies of different B cell subsets were examined.

[0304] After administration with SAR113244 and placebo, several B cell subsets were examined by flow cytometry. Frequencies were determined from lymphocytes expressing CD20. For naive B cells (CD19+CD27-IgD+), the mean percentage ranged from approximately 59.8% to 64.7% across all post-baseline time points (baseline ranged from 62.0% to 64.5%). Memory B cells before switching ( For CD19+CD27+IgD+, the mean percentage ranged from approximately 6.0% to 8.1% (baseline ranged from 6.5% to 7.9%). For switched memory B cells (CD19+CD27+IgD-), the mean percentage ranged from approximately 19.8% to 23.3% (baseline ranged from 19.8% to 21.5%). For double-negative memory B cells (CD19+CD27-IgD-), the mean percentage ranged from approximately 7.4% to 10.6% (baseline ranged from 8.7% to 9.2%).

[0305] The baseline percentages of CXCR5-expressing cells in each subset were as follows: naive B cells were 98.4% to 99.2%; pre-switch memory B cells were 98.1% to 99.1%; post-switch memory B cells were 9 4.1% to 94.2%; and double-negative memory B cells were 63.5% to 79.2%.

[0306] After administration of SAR113244 on Day 1, a decrease in CXCR5 not occupied by SAR113244 was observed on Day 8 (first visit after dosing) in the 500 mg SAR113244 treatment group and on Day 15 (first visit after dosing) in the 250 mg SAR113244 treatment group. Maximum occupancy continued until Day 85 (last available time point) in the 500 mg SAR113244 treatment group and until Day 43 in the 250 mg SAR113244 treatment group; by Day 85, RO appeared to return to baseline levels.

[0307] After administration of 250 mg Q4W SAR113244 on day 1 to CD19-expressing lymphocytes, the frequency of CD19+CD20+ cells decreased by day 15 (baseline: 94.6%, day 15: 81.0%), then gradually increased from day 15 to day 85 (day 29: 84.3%, day 43: 86.7%), and appeared to return to a level similar to that observed at baseline (day 85: 92.0%). The frequency of CD19+CD20-CD27++ cells (antibody-secreting cells) transiently increased in some patients after administration of 250 mg Q4W SAR113244 on day 1 and appeared to return to a level similar to that observed at baseline by day 85. These B cell subsets did not appear to be consistently affected by SAR113244 in the 500 mg SAR113244 treatment group.

[0308] T cell subsets The frequencies of total T cells and T cell subsets were determined.

[0309] After administration of SAR113244 and placebo, several T cell subsets were examined by flow cytometry. Frequencies were determined from CD3-expressing lymphocytes. For helper T cells (CD4+), the mean percentages ranged from approximately 51.4% to 70.5% over all post-baseline time points (baseline ranged from 53.6% to 69.5%). For cytotoxic T cells (CD8+), the mean percentages ranged from approximately 21.0% to 38.8% over all post-baseline time points (baseline ranged from 22.1% to 38.9%).

[0310] For naive T cells (CD45RA+CCR7+), the mean percentage of CD4+ cells ranged from approximately 41.9% to 50.5% over all post-baseline time points (baseline ranged from 44.8% to 49.0%); the mean percentage of CD8+ cells ranged from approximately 38.8% to 53.3% over all post-baseline time points (baseline ranged from 45.8% to 52.4%).

[0311] For central memory T cells (CD45RA−CCR7+), the mean percentage of CD4+ cells ranged from approximately 25.0% to 30.0% across all post-baseline time points (baseline range: 24.9% to 28.1%); the mean percentage of CD8+ cells ranged from approximately 3.3% to 8.5% across all post-baseline time points (baseline range: 3.6% to 9.0%).

[0312] For effector memory T cells (CD45RA−CCR7−), the mean percentage of CD4+ cells ranged from approximately 19.9% to 26.6% across all post-baseline time points (baseline range: 21.4% to 26.9%); the mean percentage of CD8+ cells ranged from approximately 18.2% to 35.2% across all post-baseline time points (baseline range: 16.1% to 28.7%).

[0313] The mean percentage of CD4+ cells expressing CXCR5 at baseline ranged from approximately 8.7% to 17.2%.

[0314] Pharmacodynamic Conclusions Saturation of CXCR5 by SAR113244 was observed at day 7 after the first dose in all patients at 250 mg and 500 mg SAR113244. For the second dose, the duration of saturation appeared to increase to a median of 42 days at 250 mg and 56 days at 500 mg. For both dosing groups, the normalized RO% decreased from the saturation zone by day 113 in 10 / 12 patients (<LLOQ in some patients), although >80% normalized RO% was still observed in two patients at day 113.

[0315] ​SAR113244 had no consistent effect on disease activity and QoL scales, serum CXCL13, autoantibody levels, complement levels, or B-cell or T-cell subsets. A transient increase in antibody-secreting cells was confirmed in some patients after administration of 250 mg of SAR113244, but a similar increase was not observed after administration of 500 mg of SAR113244.

Example

[0316] Safety evaluation Degree of exposure All patients except patient number 276001003 (receiving 250 mg of SAR113244) and patient number 276001019 (receiving 500 mg of SAR113244) received the intended dose of SAR113244 or placebo. Patient number 276001003 discontinued consent after the first dose for personal reasons and received only a single dose of 250 mg of SAR113244 on day 1. Patient number 276001019 also discontinued consent after the first dose and received only a single dose of placebo on day 1.

[0317] All other patients received two doses of SAR113244 or placebo on days 1 and 29 as intended (Table 15).

[0318]

Table 23

[0319] Adverse events Overview of adverse events Overall, 16 / 21 patients (3 / 5 patients receiving placebo, 4 / 6 patients receiving 250 mg of SAR113244, and 9 / 10 patients receiving 500 mg of SAR113244) experienced at least one TEAE (Table 16). A total of 52 TEAEs were reported throughout the trial.

[0320] There were no deaths or severe TEAEs, and there were no treatment interruptions due to TEAEs. One SAE occurred in a patient receiving placebo during the trial.

[0321] Three patients in the SAR113244 treatment group experienced AESIs of injection site erythema, and one patient in the placebo treatment group experienced an AESI of increasing ALT.

[0322]

Table 24

[0323] Display of Adverse Events Number and percentage of patients with TEAEs by treatment, main SOC, and PT were summarized in Table 17.

[0324]

Table 25

[0325]

Table 26

[0326] Analysis of Adverse Events The most frequently reported TEAEs (reported in >2 patients) were nasopharyngitis and headache reported in all treatment groups, and injection site erythema reported only in patients receiving SAR113244. Orthostatic dizziness, treatment dizziness and nausea were each reported in 2 patients, and all other TEAEs occurred in 1 patient.

[0327] No any dose-related trends were seen in the type or occurrence of reported AEs. Except for injection site erythema, there was no treatment-related trend in the type or occurrence of reported AEs.

[0328] Of the six patients in the 250 mg SAR113244 treatment group, three (50%) reported a Grade 7 AE of injection site erythema, and two out of 10 (20%) patients in the 500 mg SAR113244 treatment group reported a Grade 3 AE of injection site erythema. Patients who received placebo did not experience injection site erythema or swelling.

[0329] In the 250 mg SAR113244 treatment group, for patients numbered 276001003 and 276001011, and in the 500 mg SAR113244 treatment group, for patient numbered 27600104, the TEAE of injection site erythema lasted for 24 hours and was thus reported as an AESI.

[0330] Death, Serious Adverse Events and Other Notable Adverse Events Death During the trial, no deaths were reported.

[0331] Serious Adverse Events During the trial, one SAE occurred.

[0332] Patient numbered 276001048 in the placebo treatment group experienced postmenopausal bleeding. Fractional curettage was performed under hysteroscopy to treat the SAE without complications. This SAE was of moderate intensity and was considered not related to the IMP. The trial treatment was not changed as a result of this SAE.

[0333] Adverse Events Leading to Discontinuation and Other Notable Adverse Events There were no TEAEs that led to discontinuation of the trial.

[0334] Patients numbered 276001003 and 276001011 in the 250 mg SAR113244 treatment group, and patient numbered 276001041 in the 500 mg SAR113244 treatment group, experienced TEAEs of injection site erythema that lasted > 24 hours. All were of mild intensity and were considered related to the IMP.

[0335] Patient number 276001048 in the placebo treatment group experienced an AESI of increasing ALT. The AE was mild in intensity and was considered not related to the IMP.

[0336] Clinical trial evaluation Red blood cells, platelets, and aggregation Overall, three patients experienced a PCSA of hematocrit compared to normal baseline values (one patient in the placebo treatment group and two patients in the 500 mg SAR113244 treatment group).

[0337] White blood cells Overall, one patient reported a post - baseline PCSA of neutrophils compared to normal baseline values in the 500 mg SAR113244 treatment group. Four patients reported a post - baseline PCSA of basophils compared to normal baseline values. (Two patients in the 250 mg SAR113244 treatment group and two patients in the 500 mg SAR113244 treatment group). One PCSA of monocytes was reported in one patient in the 500 mg SAR113244 treatment group compared to normal baseline values.

[0338] Metabolism Overall, one PCSA of creatine phosphokinase was reported in one patient in the 500 mg SAR113244 treatment group compared to normal baseline values. Two PCSAs of glucose were reported in two patients in the 500 mg SAR113244 treatment group; one patient had normal baseline values and the baseline value of one patient was lost.

[0339] Electrolytes No PCSAs of electrolytes were reported in any treatment group (Table 37).

[0340] Renal function Overall, three patients reported a PCSA of creatinine (≥30% increase from baseline) compared to normal baseline values (two patients in the placebo treatment group and one patient in the 500 mg SAR113244 treatment group).

[0341]

Table 27

[0342]

Table 28

[0343]

Table 29

[0344]

Table 30

[0345]

Table 31

[0346] Liver function Regarding liver function, no PCSA was reported in any treatment group.

[0347] Vital signs, physical findings, and other safety observations Individual clinically relevant abnormalities Overall, some PCSAs of vital sign parameters occurred during the TEAE period (including patients receiving placebo).

[0348] One patient (patient number 276001036) in the 500 mg SAR113244 treatment group had a PCSA of weight loss. On day 29, the patient's weight was 11% less than the baseline value. This was not related to any TEAE, and her weight increased at the next time point (change from baseline on day 57 was -3.7%).

[0349] There were several PCSAs regarding blood pressure values. There were none related to TEAE, and the PCSAs occurred across treatment groups. No dose- or treatment-related trends were evident in the vital sign data from baseline through day 113.

[0350]

Table 32

[0351] Electrocardiogram Individual clinically relevant abnormalities An enumeration of data for patients with PCSA after baseline regarding ECG parameters is shown in Table as follows.

[0352] Clinically important possible abnormalities in heart rate were reported only in the SAR113244 treatment group. Compared with only 1 / 6 (16.7%) of patients receiving 250 mg Q4W SAR113244 and none of the patients receiving placebo, 3 / 10 (30%) of patients receiving 500 mg Q4W SAR113244 had heart rate values >90 bpm, and a treatment- and dose-related trend was seen in the increased heart rate values. However, only 1 patient receiving 500 mg Q4W SAR113244 had an increase of ≥20 bpm from baseline.

[0353] Compared with 1 / 5 (20%) of patients receiving placebo, all 5 / 6 (83.3%) of patients receiving 250 mg Q4W SAR113244 and 5 / 10 (50.0%) of patients receiving 500 mg Q4W SAR113244 had QTcB values >450 ms. 2 / 6 (33.3%) of all patients receiving 250 mg Q4W SAR113244 and 2 / 10 (20%) of patients receiving 500 mg Q4W SAR113244 had QTcF values >450 ms. Patients receiving placebo did not have QTcF values >450 ms.

[0354] Only one patient experienced a change from baseline of >30 ms in QTcB. Patient number 276001006 in the 250 mg SAR113244 treatment group had QTcB and QTcF values of 491 ms and 460 ms, respectively, after the T3H baseline on Day 1 (Table 39). The QTcB value increased by 39 ms from baseline. This was not related to any TEAE.

[0355] No increase from baseline in QTcF >480 ms or QTc interval >60 ms was observed.

[0356]

Table 33

[0357] Morphological assessment The electrocardiogram morphological assessment is summarized in Table 24.

[0358] All individual abnormalities reported were considered not clinically significant.

[0359]

Table 34

[0360] Local tolerability at the injection site During the study, injection site pain was not reported. One out of six patients (16.7%) in the 250 mg SAR113244 treatment group experienced itching at the injection site that was reported as "difficult to perceive". experienced.

[0361] One out of six patients (16.7%) in the 250 mg SAR113244 treatment group experienced injection site edema considered to be mild. This event was reported as a TEAE.

[0362] Three (50%) of six patients in the 250 mg SAR113244 treatment group and 2 / 10 (20%) of patients in the 500 mg SAR113244 treatment group experienced TEAE of injection site erythema. All were considered mild. The TEAE of injection site erythema in patient numbers 276001003 and 276001011 (250 mg SAR113244), and patient number 276001041 (500 mg SAR113244) persisted for 24 hours each and were thus reported as AESI.

[0363] Serum immunoglobulins The mean increase in IgA levels from baseline to day 113 was 492.0 mg / L (SD 242.9) in the placebo treatment group; 370. 0 mg / L (SD 351.2) in the 250 mg SAR113244 treatment group; and 83.0 mg / L (SD 395.4) in the 500 mg SAR113244 treatment group.

[0364] The mean decrease in IgE levels from baseline to day 113 was -13.66 ku / L (SD 45.58) in the placebo treatment group; -63.87 ku / L (SD 169.93) in the 250 mg SAR113244 treatment group; and -41.81 ku / L (SD 221.27) in the 500 mg SAR113244 treatment group.

[0365] The mean increase in IgM levels from baseline to day 113 was 190.0 mg / L (SD 63.6) in the placebo treatment group; 128.3 mg / L (SD 147.2) in the 250 mg SAR113244 treatment group; and 45.0 mg / L (SD 91.2) in the 500 mg SAR113244 treatment group.

[0366] SAR113244 did not appear to affect IgD or IgG.

[0367] Total peripheral blood B and T cells Since whole peripheral blood B and T cells were not measured due to the difficulty of manipulation, no data were obtained for cohort 1 (250 mg SAR113244) after baseline.

[0368] Changes in B cell (CD19) and T cell (CD3) levels from baseline to day 113 after administration of placebo or treatment with 500 mg SAR113244 were similar. Trends related to treatment were not evident in the frequencies of whole peripheral blood B and T cells.

[0369] Safety conclusions SAR113244 was generally safe and well tolerated when administered to male and female lupus patients at doses of 250 mg and 500 mg Q4W as two injections.

[0370] The most frequently reported TEAEs (reported in > 2 patients) were nasopharyngitis and headache reported in all treatment groups and injection site erythema reported only in patients receiving SAR113244. Orthostatic dizziness, treatment-related dizziness and nausea were reported in 2 patients each, and all other TEAEs occurred in 1 patient. No dose-related trends were seen in the type or occurrence of reported AEs.

[0371] In both groups of patients treated with SAR113244 and placebo, there was only minor PCSA of laboratory assessments and vital signs without dose or treatment-related trends. Minor PCSA regarding heart rate and QTcF was reported only in patients receiving SAR113244. QTcF values > 480 ms or an increase in QTcF from baseline > 30 ms were not reported in any patient.

[0372] Injection site reactions such as erythema (5 patients) and pruritus (1 patient) were reported among 16 patients receiving SAR113244 regardless of dose. All events of injection site erythema were mild.

[0373] SAR113244 showed no effect on the levels of peripheral blood total B and T cells, or immunoglobulins IgA, IgD, IgE, IgG, or IgM.

Example

[0374] Pharmacokinetic evaluation All blood samples were collected within ±15% of the scheduled sampling times in the SAR113244 protocol, and when the actual times were used for PK analysis, there was no effect on the results.

[0375] The plasma concentration of SAR113244 was below the LLOQ in all pre-dose samples on Day 1 for each dosing level.

[0376] Patient number 276001019 (500 mg SAR113244 treatment group) was not included in the PK population as the concentration data was insufficient for interpretation; the patient withdrew consent and permanently discontinued the study treatment on Day 28 and did not attend the visit on Day 15.

[0377] Plasma concentration The mean (SD) SAR113244 plasma concentration-time profiles for each dosing group after the first and second doses are shown in Figures 4 and 5, respectively.

[0378] Pharmacokinetic parameters The individual SAR113244 plasma PK parameters after the first and second doses are summarized in Tables 25 and 26, respectively.

[0379]

Table 35

[0380]

Table 36

[0381] Dose proportionality assessment Individual and mean (SD) SAR113244C max and AUC 0~4w Values are shown graphically for each dosing group in Figures 6 and 7. The results of the dose proportionality analysis are shown in Table 27 .

[0382]

Table 37

[0383] Over a dose range of 2.0-fold from 250 mg to 500 mg, mean SAR113244 C max and AUC 0~4w increased to 2.23- and 2.01-fold, respectively, after the first dose and to 1.77- and 1.88-fold, respectively, after the second dose, indicating that exposure increased with dose proportionality over the range from 250 mg to 500 mg with no major deviations.

[0384] Accumulation ratio C max and AUC 0~4w The accumulation ratios are shown in Table 28.

[0385]

Table 38

[0386] After administration of two doses of SAR113244, the accumulation ratios pooled at 250 mg and 500 mg doses were max C 0~4w was 1.47 and AUC

[0387] t 1 / 2z Dose-effect assessment for Individual and mean (SD) SAR113244t after the second dose 1 / 2z Values are shown graphically for each dosing group in Figure 8. The results of the dose-effect statistical analysis are shown in Tables 29 and 30.

[0388]

Table 39

[0389]

Table 40

[0390] No significant increase in t was observed upon administration (p = 0.098). When pooled in the range of 250 mg and 500 mg, the estimated geometric mean of t was 202 hours, i.e., 8.4 days. 1 / 2z No significant increase in t was observed upon administration (p = 0.098). When pooled in the range of 250 mg and 500 mg, the estimated geometric mean of t was 202 hours, i.e., 8.4 days. 1 / 2z No significant increase in t was observed upon administration (p = 0.098). When pooled in the range of 250 mg and 500 mg, the estimated geometric mean of t was 202 hours, i.e., 8.4 days.

[0391] Variance components Within-patient and SAR113244C max and AUC 0~4w The total SDs of were shown in Table 31.

[0392]

Table 41

[0393] The total variability expressed as CV(%) was 36.6% and 39.5% for SAR113244C max and AUC 0~4w respectively, which was moderate. The within-patient variability expressed as CV(%) was 15.0% and 12.5% for C max and AUC 0~4w respectively, which was low.

[0394] Immunogenicity Drug - resistant antibodies were not detected in any of the patients receiving placebo or any of the patients before administration of SAR113244. The incidence of treatment - induced ADA in patients receiving 250 mg Q4W SAR13244 and patients receiving 500 mg Q4W SAR113244 was 66.7% (4 / 6) and 20.0% (2 / 10) respectively. Overall, treatment - induced ADA was detected in 37.5% of the patients treated with SAR113244.

[0395] Of the six patients with detectable ADA, four patients (66.7%) had persistent treatment-induced antibodies and two patients (33.3%) had transient treatment-induced ADA.

[0396] There were approximately 24 weeks between the first and last positive samples, thus patients were not classified as having transient or persistent treatment-induced antibodies. Overall, the median time to onset of the ADA response was 73.5 days and the median ADA duration was 137.0 days (range: 23 to 170 days). The measured ADA peak titers ranged from 60 to 600 IU / mL.

[0397] Pharmacokinetic Conclusions After administration of 250 to 500 mg of SC SAR113244, all true patients were quantifiably exposed to the drug at the peak plasma concentration (median) of SAR113244 at 7 days after the first dose and after the second dose.

[0398] SAR113244 exposure increased proportionally with dose after repeated dosing from 250 mg to 500 mg. For a 2.0-fold increase in dose, the mean C max and AUC 0~4w increased 2.23- and 2.01-fold, respectively, after the first dose and 1.77- and 1.88-fold, respectively, after the second dose. After administration of two doses of SAR113244, the accumulation ratios pooled at the 250 mg and 500 mg doses were 1.47 for C max and 1.50 for AUC 0~4w .

[0399] Dose had no statistically significant effect on t 1 / 2z . The geometric mean t 1 / 2z estimate pooled across the range of 250 mg to 500 mg was 202 hours, or 8.4 days.

[0400] The total variability of SAR113244 C max and AUC 0~4w was moderate at 36.6% and 39.5%, respectively. C maxand AUC 0~4w Intra-patient variability of 0~4w was low at 15.0% and 12.5% respectively.

[0401] Overall, treatment-induced ADA was detected in 37.5% of patients treated with SAR113244. There was no dose effect on ADA. All patients (4 / 4) with treatment-induced ADA in the 250 mg SAR113244 treatment group showed a persistent ADA response, and all patients (2 / 2) with treatment-induced ADA in the 500 mg SAR113244 treatment group showed a transient ADA response.

[0402] Discussion and overall conclusions SAR113244 was generally safe and well tolerated when administered to male and female lupus patients as two injections at doses of 250 g and 500 mg Q4W.

[0403] The most frequently reported TEAEs (reported in > 2 patients) were nasopharyngitis and headache reported in all treatment groups, and injection site erythema reported only in patients receiving SAR113244. Orthostatic dizziness, treatment-related dizziness and nausea were reported in 2 patients each, and all other TEAEs occurred in 1 patient. No dose-related trends were seen in the type or occurrence of reported AEs.

[0404] There was a slight PCSA of test assessments and vital signs without dose- or treatment-related trends in both groups of patients treated with SAR113244 and placebo. A slight PCSA regarding heart rate and QTcF was reported only in patients receiving SAR113244.

[0405] Injection site reactions such as erythema (5 patients) and pruritus (1 patient) were reported among 16 patients receiving SAR113244 regardless of dose. All injection site erythema was mild.

[0406] SAR113244 showed no effect related to the levels of peripheral blood total B and T cells, or immunoglobulins IgA, IgD, IgE, IgG, or IgM.

[0407] Saturation of CXCR5 by SAR113244 occurred at day 7 after the first dose in all patients at 250 mg and 500 mg. The period of saturation for the second dose appeared to increase to a median of 42 days at 250 mg and 56 days at 500 mg. For both dosing groups, the normalized RO% decreased from the saturation zone by day 113 in 10 / 12 patients (<LLOQ in some patients), although >80% normalized RO% was still observed in two patients at day 113.

[0408] SAR113244 showed no consistent effect on disease activity and QoL scales, serum CXCL13, autoantibody levels, complement levels, or B - or T - cell subsets. A transient increase in antibody - secreting cells was confirmed in some patients after 250 mg dosing, but a similar increase was not observed after 500 mg dosing.

[0409] After administration of 250 - 500 mg SC SAR113244, all true patients were quantifiably exposed to the drug at the peak plasma concentration (median) of SAR113244 at day 7 after the first dose and after the second dose.

[0410] SAR113244 exposure increased proportionally with dose after repeated dosing from 250 mg to 500 mg. After administration of two doses of SAR113244, the accumulation ratio pooled at 250 mg and 500 mg doses was such that C max was 1.47, AUC 0~4w was 1.50. The dose showed no statistically significant effect on t 1 / 2z The pooled geometric mean t 1 / 2z estimation in the range of 250 mg to 500 mg was 202 hours, i.e., 8.4 days.

[0411] SAR113244C​max and AUC 0~4w The total variability of was moderate at 36.6% and 39.5% respectively. C max and AUC 0~4w The within-patient variability of was low at 15.0% and 12.5% respectively.

[0412] Anti-drug antibodies were not detected in any of the patients receiving placebo or in any of the patients before administration of SAR113244. Overall, treatment-induced ADA was detected in 37.5% of the patients treated with SAR113244.

[0413]

Table 42

[0414]

Table 43

[0415]

Table 44

[0416]

Table 45

[0417]

Table 46

[0418]

Table 47

[0419] Table 38 shows the number of patients with PCSA of liver function during the TEAE period.

[0420]

Table 48

[0421]

Table 49

[0422]

Table 50

[0423]

Table 51

[0424]

Table 52

[0425]

Table 53

[0426] The CDR sequences are shown in bold in the sequences of Table 40. The sequences and descriptions shown in Table 40 correspond to those disclosed in U.S. Patent No. 8,647,622 B1, which is incorporated by reference in its entirety.

Claims

1. 1. A pharmaceutical composition for treating a patient with lupus, comprising a therapeutically effective amount of an antibody or fragment thereof that specifically binds to the extracellular domain of human CXCR5, the antibody or fragment thereof comprising: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; (b) light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLAS (SEQ ID NO: 59), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively: (c) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16; (d) light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLAS (SEQ ID NO: 64), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; (e) light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSNLAS (SEQ ID NO: 65), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; (f) a variable light chain (V) comprising the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21 L ), and a variable heavy chain (V H ); (g) a variable amino acid sequence comprising the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO:

32. a light chain and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34; (h) light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RMSNLA (SEQ ID NO: 66), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; (i) light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSNLA (SEQ ID NO: 67), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; (j) light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 consisting of the amino acid sequences RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 consisting of the amino acid sequences GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; (k) a variable light chain comprising the amino acid sequence of SEQ ID NO: 35, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 37; (l) a variable light chain comprising the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 41, or SEQ ID NO: 43, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 47; (m) a variable light chain comprising the amino acid sequence of SEQ ID NO: 55, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57; or (n) light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 consisting of the amino acid sequences of RSSKSLLHSSGKTYLYW (SEQ ID NO: 69), RMSNLA (SEQ ID NO: 66), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 consisting of the amino acid sequences of GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; The pharmaceutical composition, wherein the patient has tested positive for antinuclear antibodies at a titer of >= 1:

160.

2. A pharmaceutical composition for treating a patient with lupus, comprising a therapeutically effective amount of an antibody or fragment thereof that specifically binds to the extracellular domain of human CXCR5, wherein the antibody or fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 32 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33; The pharmaceutical composition, wherein the patient has tested positive for antinuclear antibodies at a titer of >= 1:

160.

3. A pharmaceutical composition for treating a patient with lupus, comprising a therapeutically effective amount of an antibody or fragment thereof that specifically binds to the extracellular domain of human CXCR5, wherein the antibody or fragment thereof comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 consisting of the amino acid sequences of RSSKSLLHSSGKTYLY (SEQ ID NO: 58), RLSSLA (SEQ ID NO: 68), and MQHLEYPYT (SEQ ID NO: 60), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 consisting of the amino acid sequences of GFSLIDYGVN (SEQ ID NO: 61), VIWGDGTTY (SEQ ID NO: 62), and IVY (SEQ ID NO: 63), respectively; The patient has tested positive for antinuclear antibodies at a titer of ≧1:

160. Finished product.

4. The pharmaceutical composition of claim 1 , 2, or 3, wherein the antibody or fragment thereof further comprises one or more constant region domains.

5. The antibody or fragment thereof is H1 , C H2 , C H3 4. The pharmaceutical composition of claim 1, 2, or 3, further comprising:

6. The pharmaceutical composition of claim 4 , wherein one or more constant region domains are derived from an IgG antibody.

7. The pharmaceutical composition of claim 6 , wherein the IgG antibody is an IgG4 antibody.

8. The pharmaceutical composition of claim 1 , 2 or 3 , wherein the antibody or fragment thereof is a single-chain Fv antibody.

9. 4. The pharmaceutical composition of claim 1, 2, or 3, wherein the antibody or fragment thereof is administered subcutaneously to the patient.

10. 4. The pharmaceutical composition of claim 1, 2, or 3, wherein the antibody or fragment thereof is administered at a concentration of 500 mg.

11. 4. The pharmaceutical composition of claim 1, 2, or 3, wherein the patient has an anti-dsDNA antibody titer of at least 11 IU / mL.

12. 4. The pharmaceutical composition of claim 1, 2, or 3, wherein the patient has a systemic lupus erythematosus disease activity index of at least 4.

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