Anti-WISP1 Antibody and Method of Use

Abstract

Antibodies and antibody fragments that specifically bind to WNT1-induced signaling pathway protein 1 (WISP1) are provided herein. The anti-WISP1 antibody provided herein binds to domain 2 of the WISP1 protein and inhibits WISP1 protein activity. The anti-WISP1 antibody is considered effective for treating and preventing fibrosis in subjects requiring treatment and prevention of fibrosis. In an embodiment, the fibrosis is hepatic fibrosis or pulmonary fibrosis.

Description

Background Technology

[0001] Cross-reference regarding related applications

[0002] This application claims priority to U.S. Provisional Application No. 63,625,877 filed January 26, 2024, the full text of which is incorporated herein by reference for all purposes.

[0003] Reference to the electronic sequence list

[0004] The contents of the electronic sequence list (062698-505001WO_Sequence_Listing_ST26.xml; size: 28,595 bytes; and creation date: January 23, 2025) are incorporated herein by reference in their entirety.

[0005] Background Technology

[0006] While the formation of fibrous tissue is part of the normal, beneficial process of healing after injury, fibrosis is a pathological condition characterized by the abnormal accumulation of collagen matrix following injury or inflammation, which alters the structure and function of various tissues. Progressive fibrosis in the kidneys, liver, lungs, heart, bones or bone marrow, and skin is a major cause of death. Many diseases associated with the proliferation of fibrous tissue are chronic and often debilitating. Some, including pulmonary fibrosis, are fatal, partly due to the fact that currently available treatments have severe side effects and are generally ineffective in slowing or stopping the progression of fibrosis.

[0007] WNT1-induced signaling pathway protein 1 (WISP1) expression is a characteristic of fibrosis progression. WISP1 contains four domains, and its expression causes abnormal myofibroblast accumulation and excessive collagen deposition in fibrotic disorders.

[0008] In one embodiment, an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody is provided, comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and the light chain variable domain comprises CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0009] In one embodiment, an anti-WISP1 antibody is provided, wherein the anti-WISP1 antibody binds to the same epitope as the anti-WISP1 antibody comprising a heavy chain variable domain including CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain including CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0010] In another embodiment, an antibody competing with an anti-WISP1 antibody for binding to a WISP1 protein is provided, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0011] In another embodiment, an anti-WNT1-induced-signaling pathway protein 1 (WISP1) antibody is provided, wherein the antibody comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDR H1, CDR H2, and CDR H3, the light chain variable domain comprises CDR L1, CDR L2, and CDR L3, CDR H1 is composed of sequence number 1, CDR H2 is composed of sequence number 2, CDR H3 is composed of sequence number 3, CDR L1 is composed of sequence number 4, CDR L2 is composed of sequence AAS, and CDR L3 is composed of sequence number 6.

[0012] In another embodiment, an anti-WNT1-induced-signaling pathway protein 1 (WISP1) antibody is provided, wherein the antibody binds to the same epitope as the anti-WISP1 antibody comprising a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDR H1, CDR H2, and CDR H3, the light chain variable domain comprises CDR L1, CDR L2, and CDR L3, CDR H1 is composed of sequence number 1, CDR H2 is composed of sequence number 2, CDR H3 is composed of sequence number 3, CDR L1 is composed of sequence number 4, CDR L2 is composed of sequence AAS, and CDR L3 is composed of sequence number 6.

[0013] In another embodiment, an antibody competing with an anti-WISP1 antibody for binding to a WISP1 protein is provided, wherein the anti-WISP1 antibody comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDR H1, CDR H2, and CDR H3, the light chain variable domain comprises CDR L1, CDR L2, and CDR L3, CDR H1 is composed of sequence number 1, CDR H2 is composed of sequence number 2, CDR H3 is composed of sequence number 3, CDR L1 is composed of sequence number 4, CDR L2 is composed of sequence AAS, and CDR L3 is composed of sequence number 6.

[0014] In one embodiment, a method for treating fibrosis in a subject requiring treatment for fibrosis is provided, said method comprising the step of administering to said subject a therapeutically effective amount of the antibody provided herein, which includes an embodiment thereof.

[0015] In one embodiment, a method for treating fibrosis in a subject requiring treatment for fibrosis is provided, said method comprising the following steps: a) identifying, in a biological sample obtained from said subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) a step of administering a therapeutically effective amount of the anti-WISP1 antibody provided herein, including an embodiment thereof, to the subject.

[0016] In one embodiment, a method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell is provided, said method comprising the step of contacting a cell with an anti-WISP1 antibody provided herein, which includes an embodiment thereof.

[0017] In one embodiment, a method for inhibiting the migration of WISP-1 receptor-expressing cells is provided, said method comprising the step of contacting said cells with an anti-WISP1 antibody provided herein, which includes an embodiment thereof.

[0018] A method for detecting WISP1-expressing cells comprises: i) contacting WISP1-expressing cells with an antibody provided herein, comprising an embodiment thereof; and ii) detecting the binding of the antibody to a WISP1 protein expressed by the cells. Brief explanation of the drawing

[0019] Fig. 1. Enzyme-linked immunosorbent assay (ELISA) results showing hybridoma-expression antibody binding to recombinant human WISP1 protein. Fig. 2. Analysis of the binding specificity of isolated anti-WISP1 antibody clones against human WISP1 protein. Fig. 3. SPR data showing the binding kinetics of isolated anti-WISP1 antibody clones against human WISP1. Fig. 4. Binding of anti-WISP1 antibody clones to different domains of WISP1. Figs. 5a-5b. In vivo efficacy study of anti-WISP1 antibody in carbon tetrachloride (CCL4) liver fibrosis model. Fig. 5a ) Data illustrating that anti-WISP1 inhibits liver fibrosis, as measured by a decrease in hydroxyproline and signs of reduced collagen content in fibrous tissue. Data are expressed as mean + / - SEM; *p<0.05, one-way ANOVA using Dunnett's multiple comparison versus +PBS. Fig. 5b) The results show that the anti-WISP1 antibody K4 reduces hydroxyproline by up to 76.5% compared to the vehicle control, and the anti-WISP1 antibody M6-QYS reduces hydroxyproline by up to 79.7%. Figs. 6a-6b. In vivo efficacy studies using anti-WISP1 antibodies in a bile duct ligation (BDL) liver fibrosis model exemplify that M6-QYS and K4 alleviated the BDL-induced increase in liver α-SMA. Fig. 6a)Administration of M6-QYS and K4 reduced α-SMA in a BDL liver fibrosis mouse model compared to a vehicle-treated control group. Fig. 6b) Representative image of an α-smooth muscle actin (α-SMA) stained histological section obtained at 100X magnification. Darker counterstaining indicates positive staining for α-SMA. The image indicates that BDL increased hepatic α-smooth muscle actin (α-SMA) compared to sham treatment. Fig. 7. Anti-WISP1 antibody K4 reduces pSMAD2 in normal rat renal fibroblasts treated with full-length (FL) WISP1. The inhibition of pSMAD2 by anti-WISP1 antibodies varied from 0% to 71%. Monoclonal antibody E8 reduced pSMAD2 by 44%, while antibody K4 reduced the signal by 63%, similar to the reduction in pSMAD2 (65%; positive control) after treatment with the polyclonal anti-WISP1 AF1627 antibody. Fig. 8. Anti-WISP1 K4 antibodies reduce primary human hepatic astrocyte (HHSC) chemotaxis / motility. Starting with a concentration of 2 μg / mL, monoclonal anti-WISP1 antibody K4 inhibited chemotaxis and increased inhibition at higher doses (lower columns). Treatment with the polyclonal anti-WISP1 antibody R&D AF1627 (positive control) also reduced WISP1-induced chemotaxis starting at 2 μg / mL (middle column). IgG antibody treatment did not alter the chemotaxis rate compared to wells without antibodies, even at the highest concentration evaluated (top column). Leukocytes are transferred from the top of the transwell plate to the bottom chamber. Fig. 9.Anti-WISP1 K4 antibodies reduce primary human hepatic astrocyte (HHSC) chemotaxis / motility. HHSC migration at 48 h was quantified. The conditions were as described in Figure 8. Chemotaxis was calculated using the "Phase Area Bottom" metric, normalized to the initial peak. Significant HHSC chemotaxis occurred with the combination of WISP1 and 10% FBS, but not in the absence of FBS. Non-specific IgG antibodies did not inhibit chemotaxis / cell migration even at the peak concentration (100 μg / mL). The polyclonal anti-WISP1 antibody AF1627 inhibited chemotaxis starting at 2 μg / mL, and inhibition increased at higher antibody concentrations. The monoclonal anti-WISP1 antibody K4 also significantly inhibited chemotaxis starting at 2 μg / mL, and inhibition increased with antibody concentration. Significance is indicated for IgG at 100 μg / mL using 10% FBS (p < 0.05). Fig. 10. IC50 for anti-WISP1 antibody K4. IC50 and IC80 values ​​were determined using a Python-based nonlinear regression curve fit to dose-response data from the final time point (48h). The Mediar monoclonal anti-WISP1 antibody K4 produced an IC50 of 1.8788 μg / mL and an IC80 of 4.7959 μg / mL (left panel); the polyclonal anti-WISP1 antibody AF1627 had an IC50 of 1.6629 μg / mL and an IC80 of 5.152 μg / mL (right panel). Specific details for implementing the invention

[0020] Various embodiments and aspects of the present invention are set forth and described herein, and it will be obvious to those skilled in the art that such embodiments and aspects are provided merely for illustrative purposes. Various modifications, variations, and substitutions will be conceived by those skilled in the art without departing from the invention. It should be understood that various alternative examples of the embodiments of the invention described herein may be used to practice the invention.

[0021] Section titles used herein are for organization purposes only and should not be construed as limiting the subject matter described. Any literature cited in applications, including patents, patent applications, articles, books, manuals, and papers, without limitation, or any part of such literature, is explicitly incorporated herein by reference in its entirety for any purpose.

[0022] The abbreviations used herein have their conventional meanings in the fields of chemistry and biology. The chemical structures and formulas presented herein are interpreted according to the standard rules of chemical valence known in the field of chemistry.

[0023] Unless otherwise defined, technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art. For example, references are made to [Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY 2nd ed., J. Wiley & Sons (New York, NY 1994)] and [Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989)], the full text of which is incorporated herein for all purposes. Any method, apparatus, or material similar or identical to that described herein may be used in the practice of the invention. The following definitions are provided to aid in understanding specific terms frequently used herein and are not intended to limit the scope of the disclosure.

[0024] As used herein, the term “approx.” means a range of values ​​including a specific value, which a person skilled in the art will consider to be reasonably similar to the specific value. In embodiments, the term “approx.” means being within a standard deviation using a measurement method generally acceptable in the art. In embodiments, “approx.” means a range extending up to + / - 10% of a specific value. In embodiments, “approx.” means a specific value.

[0025] As used herein, the terms “includes” or “comprising” are used to indicate that the following list of components or steps is incomplete. This implies that additional components or steps may be present in the invention even if they are not explicitly mentioned. The terms “includes” or “including” are synonymous with “comprising” and are considered open-ended, allowing for the inclusion of other unmentioned components or steps.

[0026] As used herein, the terms "is accomplished" or "is accomplished" are used as closed terms excluding any components, steps, or elements not specifically mentioned after the term. The use of the terms "is accomplished" or "is accomplished" implies that the mentioned features are limited to these elements alone and do not permit the inclusion of any additional elements.

[0027] "Nucleic acid" refers to a nucleotide (e.g., deoxyribonucleotide or ribonucleotide) in a single-, double-, or multi-stranded form and a polymer thereof; or a nucleoside (e.g., deoxyribonucleotide or ribonucleotide). In the embodiments, "nucleic acid" does not include a nucleoside. The terms "polynucleotide," "oligonucleotide," "oligo," etc., refer to a linear sequence of nucleotides in the ordinary conventional sense. The term "nucleoside" refers to a glycosylamine comprising a nucleobase and a pentose sugar (ribose or deoxyribose) in the ordinary conventional sense. Non-limiting examples of nucleosides include cytidine, uridine, adenosine, guanosine, thymidine, and inosine. The term "nucleotide" refers to a single unit of a polynucleotide, i.e., a monomer, in the ordinary conventional sense. A nucleotide may be a ribonucleotide, a deoxyribonucleotide, or a modified form thereof. Examples of polynucleotides considered herein include single- and double-stranded DNA, single- and double-stranded RNA, and hybrid molecules having mixtures of single- and double-stranded DNA and RNA. Nucleic acids, for example, examples of polynucleotides considered herein include any type of RNA, e.g., mRNA, siRNA, miRNA, and guide RNA, and any type of DNA, genomic DNA, plasmid DNA, and minicircle DNA, and any fragments thereof. In the context of polynucleotides, the term "duplex" refers to a double strand in the ordinary conventional sense. Nucleic acids may be linear or branched. For example, a nucleic acid may be a linear chain of nucleotides, or a nucleic acid may be branched such that, for example, the nucleic acid contains one or more arms or branches of nucleotides. Optionally, branched nucleic acids are repeatedly branched to form higher-order structures, such as dendrimers, etc.

[0028] For example, nucleic acids comprising a nucleic acid having a phosphothioate backbone may comprise one or more reactive moietys. As used herein, the term reactive moiety comprises any group capable of reacting with other molecules, e.g., nucleic acids or polypeptides, through covalent, non-covalent, or other interactions. As an example, the nucleic acid may comprise an amino acid reactive moiety that reacts with amino acids on a protein or polypeptide through covalent, non-covalent, or other interactions.

[0029] The above terms also encompass known nucleotide analogs or non-naturally occurring nucleic acids containing modified backbone residues or linkages, which have binding properties similar to the reference nucleic acid and are metabolized in a manner similar to the reference nucleotide. Examples of such analogs include, without limitation, phosphoramidates, phosphorodiamidates, phosphothioates (also known as phosphothioates in which the oxygen of a phosphate is replaced by a double-bonded sulfur), phosphodithioates, phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, boron phosphonate, or phosphodiester derivatives comprising O-methylphosphoroamidite linkages (see Eckstein, OLIGONUCLEOTIDES AND ANALOGUES: A PRACTICAL APPROACH, Oxford University Press, the full text of which is incorporated herein for all purposes), as well as modifications to nucleotide bases such as 5-methyl cytidine or pseudouridine; and peptide nucleic acid backbones and linkages. Other analog nucleic acids include those having a positively charged backbone; a nonionic backbone, a modified sugar, and a non-ribose backbone (e.g., phosphorodiamidate morpholino oligos or lock nucleic acids (LNAs) as known in the art), including those described in U.S. Patents No. 5,235,033 and No. 5,034,506, the full text of which is incorporated herein for all purposes, and in the literature [Chapters 6 and 7, ASC Symposium Series 580, CARBOHYDRATE MODIFICATIONS IN ANTISENSE RESEARCH, Sanghui & Cook, eds]. Nucleic acids comprising one or more carbocyclic sugars are included within one definition of nucleic acid. Modification of the ribose-phosphate backbone may be performed for various reasons, for example, to increase the stability and half-life of these molecules in a physiological environment or as probes on a biochip.A mixture of naturally occurring nucleic acids and analogs may be prepared; alternatively, a mixture of different nucleic acid analogs and a mixture of naturally occurring nucleic acids and analogs may be prepared. In an embodiment, the internucleotide linkage in DNA is a phosphodiester, a phosphodiester derivative, or a combination of both.

[0030] Nucleic acids may contain non-specific sequences. As used herein, the term "non-specific sequence" refers to a nucleic acid sequence containing a series of residues that are not designed to be complementary or merely partially complementary to any other nucleic acid sequence. For example, a non-specific nucleic acid sequence is a sequence of nucleic acid residues that does not function as an inhibitory nucleic acid when in contact with a cell or organism.

[0031] Polynucleotides typically consist of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (uracil (U) for thymine (T) if the polynucleotide is RNA). Thus, the term "polynucleotide sequence" is an alphabetic representation of a polynucleotide molecule; alternatively, the term may be applied to the polynucleotide molecule itself. Such an alphabetic representation may be entered into a database of a computer with a central processing unit and may be used in bioinformatics applications such as functional genomics and homology search. Polynucleotides may optionally include one or more non-standard nucleotide(s), nucleotide analog(s), and / or modified nucleotides.

[0032] As used herein, the term “complement” refers to a nucleotide (e.g., RNA or DNA) or a sequence of nucleotides capable of forming base pairs with a complementary nucleotide or sequence of nucleotides. As described herein and generally known in the art, the nucleotide complementary (matching) to adenosine is thymidine, and the nucleotide complementary (matching) to guanosine is cytosine. Accordingly, the complement comprises a sequence of nucleotides that form base pairs with the corresponding complementary nucleotide of the second nucleic acid sequence. The nucleotides of the complement may be partially or completely matched with the nucleotides of the second nucleic acid sequence. When the nucleotides of the complement are completely matched with each nucleotide of the second nucleic acid sequence, the complement forms base pairs with each nucleotide of the second nucleic acid sequence. When a nucleotide of complement partially matches a nucleotide of a second nucleic acid sequence, only some of the nucleotides of the complement form base pairs with the nucleotide of the second nucleic acid sequence. Examples of complementary sequences include coding and non-coding sequences, where the non-coding sequence contains a nucleotide complementary to the coding sequence and thus forms the complement of the coding sequence. Additional examples of complementary sequences are sense and antisense sequences, where the sense sequence contains a nucleotide complementary to the antisense sequence and thus forms the complement of the antisense sequence.

[0033] As described herein, sequence complementarity may be partial when only a portion of the nucleic acids match according to base pairing, or total when all nucleic acids match according to base pairing. Thus, two sequences that are complementary to each other may have a specific percentage of identical nucleotides (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity across a specific region).

[0034] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as amino acids subsequently modified, Examples include hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds having the same basic chemical structure as naturally occurring amino acids, namely, an α-carbon bonded to a hydrogen, carboxyl group, amino group, and R group, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. These analogs possess a modified R group (e.g., norleucine) or a modified peptide backbone, but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimics refer to chemical compounds that have a structure different from the general chemical structure of amino acids but function in a manner similar to naturally occurring amino acids. The terms "non-naturally occurring amino acids" and "non-natural amino acids" refer to amino acid analogs, synthetic amino acids, and amino acid mimics that are not found in nature.

[0035] Amino acids can be represented by their commonly known three-character symbols or by the one-character symbols recommended by the IUPAC-IUB Biochemical Nomenclature Committee. Nucleotides, similarly, can be represented by their generally accepted single-character codes.

[0036] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acid residues, wherein, in embodiments, the polymer may be conjugated to a moiety not composed of amino acids. The terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. “Fusion protein” refers to a chimeric protein encoding two or more distinct protein sequences expressed in a recombination manner as a single moiety.

[0037] The amino acid or nucleotide base "position" is indicated by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence based on its position relative to the N-terminus (or 5'-terminus). Due to deletions, insertions, cleavages, fusions, etc., that must be considered when determining optimal alignment, the number of amino acid residues in the test sequence, which is generally determined by simply counting from the N-terminus, will not necessarily be equal to the number of corresponding positions in the reference sequence. For example, if a variant has a deletion in the aligned reference sequence, there will be no amino acid in the variant corresponding to a position in the reference sequence at the deletion site. If an insertion exists in the aligned reference sequence, that insertion will not correspond to a numbered amino acid position in the reference sequence. In the case of cleavage or fusion, there may be a stretch of amino acids in the reference or aligned sequence that does not correspond to any amino acid in the corresponding sequence.

[0038] When used in the context of numbering a given amino acid or polynucleotide sequence, the terms “numbered by reference” or “corresponding to” relate to the numbering of residues of a specified reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence. An amino acid residue in a protein “corresponds” to a given residue when it occupies the same essential structural position within the protein as the given residue. Those skilled in the art will immediately recognize the type and position of the residue corresponding to a specific position in one protein (e.g., WISP1) in other proteins using different numbering systems. For example, by performing a simple sequence alignment on the protein (e.g., WISP1), the type and position of the residue corresponding to a specific position in the protein are identified in other protein sequences aligned with the protein. For example, a selected residue in a selected protein corresponds to glutamate at position 138 when the selected residue occupies the same essential space or other structural relationship as glutamate at position 138. In some embodiments, when a selected protein is aligned for maximum homology with the protein, the position in the aligned selected protein that aligns with glutamate 138 corresponds to glutamate 138. Instead of primary sequence alignment, three-dimensional structural alignment may be used, for example, when the structure of the selected protein is aligned for maximum match with glutamate at position 138 and the entire structures are compared. In this case, the amino acid occupying the same essential position as glutamate 138 in the structural model corresponds to the glutamate 138 residue.

[0039] The term "conservatively modified variant" applies to both amino acids and nucleic acid sequences. With respect to a specific nucleic acid sequence, a "conservatively modified variant" refers to a nucleic acid that codes for the same or essentially identical amino acid sequence. Due to the degeneration of the genetic code, multiple nucleic acid sequences will code for any given protein. For example, codons GCA, GCC, GCG, and GCU all code for the amino acid alanine. Therefore, at any position where alanine is specified by a codon, the codon may be changed to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silencing variations," which are a type of conservatively modified variant. All nucleic acid sequences in this document that code for polypeptides also describe all possible silencing variations of the nucleic acid. Those skilled in the art will recognize that each codon in the nucleic acid (except AUG, which is generally the only codon for methionine, and TGG, which is generally the only codon for tryptophan) can be modified to produce a functionally identical molecule. Therefore, each silencing variant of the nucleic acid encoding the polypeptide is contained in the respective described sequence.

[0040] With respect to amino acid sequences, those skilled in the art will recognize that individual substitutions, deletions, or additions to nucleic acid, peptide, polypeptide, or protein sequences alter, add, or delete a single amino acid, or that a small percentage of amino acids in the coded sequence are "conservatively modified variants," wherein the modification results in the substitution of an amino acid with a chemically similar amino acid. Tables of conservative substitutions providing functionally similar amino acids are well known in the art. These conservedly modified variants are, but are not excluded from, polymorphic variants of the initiation content, interspecies homologues, and alleles.

[0041] Each of the following eight groups contains an amino acid that is a conservative substitution for each other:

[0042] 1) Alanine (A), Glycine (G);

[0043] 2) Aspartic acid (D), glutamic acid (E);

[0044] 3) Asparagine (N), Glutamine (Q);

[0045] 4) Arginine(R), Lysine(K);

[0046] 5) Isoleucine (I), leucine (L), methionine (M), valine (V);

[0047] 6) Phenylalanine (F), tyrosine (Y), tryptophan (W);

[0048] 7) Serine (S), Threonine (T); and

[0049] 8) Cysteine ​​(C), Methionine (M)

[0050] (For example, literature [Creighton, Proteins See (1984)]).

[0051] In the embodiments, amino acid sequence variants of the antibody provided herein (e.g., antibody variants of the anti-WISP1 antibody comprising one or more amino acid residue modifications and their antigen-binding fragments) are considered. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of the antibody may be prepared by introducing appropriate modifications to the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions of the antibody's amino acid sequence, and / or insertions into it and / or substitutions of residues therein. Any combination of deletions, insertions, and substitutions may be made to obtain a final construct, provided that the final construct has a desired feature, e.g., antigen binding.

[0052] In an embodiment, an antibody variant having one or more amino acid substitutions is provided. The sites of interest for substitution mutagenicity include CDR and FR. Such variants may be "conservatively modified variants," wherein the substitution comprises exchanging one or more amino acids for other amino acids from the same group as presented above. Such variants may be "non-conservatively modified variants," wherein the substitution comprises exchanging one or more amino acids for other amino acids from a different group than presented above.

[0053] Additionally, any natural residue in the polypeptide may also be substituted with alanine, as previously described for alanine scanning mutagenesis (MacLennan et al. (1998) Acta Physiol Sc and Suppl 643: 55-67; ​​Sasaki et al. (1998) Adv Biophys 35: 1-24). Amino acid substitutions for the antibody of the invention may be prepared by a method known, for example, by PCR mutagenesis (U.S. Patent No. 4,683,195). The amino acid substitutions may be injected into the antibody of interest, and the product is screened for desired activities, such as, for example, maintained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0054] In an embodiment, the antibody provided herein is modified to increase or decrease the range in which the antibody is glycosylated. The addition or deletion of glycosylation sites for the antibody can be conveniently achieved by modifying the amino acid sequence so that one or more glycosylation sites are created or removed.

[0055] If the antibody contains an Fc region, the carbohydrate attached thereto may be modified. Natural antibodies produced by mammalian cells typically contain a branched biantennari oligosaccharide generally attached by an N-link to Asn297 of the CH2 domain of the Fc region. For example, see the literature [Wright et al., TIBTECH 15:26-32 (1997)]. The oligosaccharide may include various carbohydrates such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to GlcNAc in the "stem" portion of the biantennari oligosaccharide structure. In some embodiments, modification of the oligosaccharide in the antibody of the invention may be made to produce antibody variants having improved properties.

[0056] In one embodiment, the antibody variant is provided to have a fucose-deficient carbohydrate structure attached (directly or indirectly) to the Fc region. For example, the amount of fucose in such antibodies may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain in Asn297 for the sum of all glycostructures attached to Asn 297 (e.g., complexes, hybrids, and higher-order mannose structures) measured by MALDI-TOF mass spectrometry as described, for example, in WO 2008 / 077546. Asn297 refers to an asparagine residue located at approximately position 297 (Eu numbering of Fc region residues) in the Fc region; However, Asn297 may also be located about ±3 amino acids upstream or downstream of position 297, i.e., at positions 294 through 300, due to a few sequence changes within the antibody. These fucosylated variants may have enhanced ADCC function. For example, refer to U.S. Patent Publication Nos. US 2003 / 0157108; US 2004 / 0093621. Examples of disclosures relating to "defucosylated" or "fucose-deficient" antibody variants include the following: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol. 336:1239-1249 (2004)]; Yamane-Ohnuki et al., Biotech. Bioeng.87: 614 (2004)]. Examples of cell lines capable of producing defucosylated antibodies are Lec13 CHO cells deficient in protein fucosylation (Reference [Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986)]; US Patent Application No. US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., particularly Example 11), and knockout cell lines, e.g., alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (e.g., Reference [Yamane-Ohnuki et al., Biotech. Bioeng. 87:614 (2004)]; Reference [Kanda et al., Biotechnol. Bioeng., 94(4):680-688 (2006)]; and Includes WO2003 / 085107.

[0057] Antibody variants are further provided to have a bisected oligosaccharide, for example, the biantennari oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878; U.S. Patent No. 6,602,684; and U.S. 2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087; WO 1998 / 58964; and WO 1999 / 22764 (Raju, S.).

[0058] In an embodiment, one or more amino acid modifications may be introduced into the Fc region of the antibody provided herein to produce an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., human IgG1, IgG2, IgG3, or IgG4 Fc region) containing amino acid residue modifications (e.g., substitutions) at one or more amino acid positions.

[0059] In an embodiment, the present invention considers antibody variants having some, but not all, effector functions, which make them desirable candidates for applications where the in vivo half-life of the antibody is important but effector functions (e.g., complement and ADCC) are unnecessary or harmful. In vitro and / or in vivo cytotoxicity assays may be performed to confirm a reduction or loss of CDC and / or ADCC activity. For example, an Fc receptor (FcR) binding assay may be performed to ensure that the antibody lacks FcγR binding (which may consequently result in a lack of ADCC activity) but retains FcRn binding ability. NK cells, which are the major cells for mediating ADCC, express only Fc(RIII), whereas monocytes express Fc(RI, Fc(RII), and Fc(RIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of the literature [Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991)].

[0060] Non-limiting examples of in vitro assays for evaluating the ADCC activity of molecules of interest are described below: U.S. Patent No. 5,500,362 (see, for example, [Hellstrom et al., Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)] and [Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985)]; U.S. Patent No. 5,821,337; and [Bruggemann et al., J. Exp. Med. 166:1351-1361 (1987)]). Alternatively, non-radioactive assay methods may be used (see, for example, the ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc., Mountain View, CA); and the CYTOTOX 96® non-radioactive cytotoxicity assay (Promega, Madison, WI)). Useful effector cells for these assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, or additionally, the ADCC activity of the molecule of interest may be evaluated in vivo, for example, in animal models such as those disclosed in the following literature: [Clynes et al., Proc. Nat'l Acad. Sci. USA 95:652-656 (1998)]. A C1q binding assay may also be performed to confirm that the antibody cannot bind to C1q and consequently lacks ADCC activity. For example, in WO 2006 / 029879 and WO 2005 / 100402 Refer to C1q and C3c binding ELISAs. To evaluate complement activation, a CDC assay may be performed (e.g., [Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996)]; [Cragg et al., Blood 101:1045-1052 (2003)]; and [Cragg et al.See [ , Blood 103:2738-2743 (2004)]). Determination of FcRn binding and in vivo clearance / half-life can also be performed using methods known in the art (e.g., see Petkova et al., Int'l. Immunol. 18(12):1759-1769 (2006)]).

[0061] Antibodies having reduced effector function include having one or more substitutions of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent No. 6,737,056). Such Fc mutants include so-called "DANA" Fc mutants having substitutions at two or more amino acid positions 265, 269, 270, 297, and 327, including substitutions at residues 265 and 297 for alanine (U.S. Patent No. 7,332,581).

[0062] Antibody variants having improved or reduced binding to FcR are described below (e.g., U.S. Patent No. 6,737,056; WO 2004 / 056312, and see Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)).

[0063] The term "identical" or "identical" percentage in the context of two or more nucleic acid or polypeptide sequences relates to two or more sequences or subsequences that are identical or have a specified percentage of amino acid residues or nucleotides as measured by using the BLAST or BLAST 2.0 sequence comparison algorithm using the basic parameters listed below, or by manual alignment and visual inspection (i.e., about 60% identity for a specific region, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identity for a specific region when compared and aligned for maximum match over a comparison window or specified region) (see, e.g., NCBI website http: / / www.ncbi.nlm.nih.gov / BLAST / etc.). These sequences are hereinafter referred to as "substantially identical." These definitions may also relate to or apply to the supplementation of the test sequence. The definitions also include sequences having deletions and / or additions as well as sequences having substitutions. As described below, the preferred algorithm may consider gaps, etc. Preferably, the identity exists over a length of at least about 25 amino acids or nucleotides, or more preferably over a length of 50 to 100 amino acids or nucleotides.

[0064] The “percentage of sequence identity” is determined by comparing two optimally aligned sequences for a comparison window, wherein a portion of the polynucleotide or polypeptide sequence in the comparison window may include additions or deletions (i.e., gaps) as compared to a reference sequence for optimal alignment of the two sequences (which does not include additions or deletions). The percentage is calculated by determining the number of positions where the same nucleic acid base or amino acid residue is generated in the two sequences to calculate the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to calculate the percentage of sequence identity.

[0065] As used herein, a “comparison window” comprises, for example, a reference to any one segment of a whole sequence or a number of consecutive positions selected from the group consisting of 20 to 600, about 50 to about 200, or about 100 to about 150 amino acids or nucleotides, wherein the sequence may be compared with the same number of reference sequences of consecutive positions after two sequences have been optimally aligned. Methods for aligning sequences for comparison are well known in the art. Optimal alignment of sequences for comparison is, for example, described in the literature [Smith and Waterman (1970)], the full text of which is incorporated herein for all purposes. Adv. Appl. Math. By the local homology algorithm of 2:482c], the literature [Needleman and Wunsch (1970) whose full text is incorporated herein for all purposes] J. Mol. Biol. By the homology alignment algorithm of 48:443], the full text of the literature [Pearson and Lipman (1988)] included herein for all purposes Proc. Nat'l. Acad. Sci. USASimilarity investigation methods of [85:2444] may be carried out by computer execution of these algorithms (GAP, BESTFIT, FASTA, and TFASTA of the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr., Madison, WI, the full text of which is incorporated herein for all purposes), or by manual alignment and visual inspection (e.g., see Ausubel et al., Current Protocols in Molecular Biology (1995 supplement)), the full text of which is incorporated herein for all purposes).

[0066] Examples of algorithms suitable for determining the percentages of sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in the following literature: Literature [Altschul et al. (1977) Nuc. Acids Res. 25:3389-3402], and literature[Altschul et al. (1990) J. Mol. Biol [ . 215:403-410], each in its entirety incorporated herein for all purposes. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). These algorithms include the step of prioritizing the identification of high-scoring sequence pairs (HSPs) by identifying short words of length W in a query sequence that match or satisfy some positive threshold score T when aligned with words of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al. , The aboveThese initial neighborhood word hits serve as seeds to initiate a search for longer HSPs containing them. Word match results are extended in two directions according to each sequence up to the limit where the cumulative alignment score can increase. For nucleotide sequences, the cumulative score is calculated using parameters M (reward score for matching residue pairs, always > 0) and N (penalty score for mismatched residues, always < 0). For amino acid sequences, the cumulative score is calculated using a score matrix. The extension of word match results in each direction is stopped in the following cases: when the cumulative alignment score decreases by X amount from its maximum reach value; when the cumulative score becomes zero or less due to the accumulation of alignments with one or more negative score residues; or when reaching the end of any of the sequences. The BLAST algorithm parameters W, T, and X determine the speed and sensitivity of the alignment. The BLASTN program (for nucleotide sequences) uses a word length (W) of 11, an expectation value (E) of 10, M=5, N=-4, and a comparison of two strands by default. For amino acid sequences, the BLASTP program uses a word length of 3, an expectation value (E) of 10, and a BLOSUM62 score matrix (reference [Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA [See 89:10910, the full text of which is incorporated herein for all purposes] Use alignment of 50 (B), expected value of 10 (E), M=5, N=-4, and comparison of the two strands.

[0067] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (e.g., literature [Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA[See 90:5873-5780, the full text of which is incorporated herein for all purposes). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indicator of the probability that a match between two nucleotide or amino acid sequences will occur by chance. For example, a nucleic acid is considered similar to a reference sequence if, in a comparison of a test nucleic acid to a reference nucleic acid, the smallest sum probability is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.

[0068] An indicator that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the antibody produced against the polypeptide encoded by the second nucleic acid as described below. Accordingly, the polypeptide is typically substantially identical to the second polypeptide, for example, when the two peptides differ only in conservative substitutions. Another indicator that two nucleic acid sequences are substantially identical is that the two molecules or their complement hybridize with each other under strict conditions as described below. Yet another indicator that two nucleic acid sequences are substantially identical is that the same primer can be used to amplify the sequences.

[0069] Antibodies are large, complex molecules with a complex internal structure (molecular weight of approximately 150,000 or about 1,320 amino acids). Natural antibody molecules contain two identical pairs of polypeptide chains, each pair having one light chain and one heavy chain. Each light chain and heavy chain ultimately consists of the following two regions: a variable ("V") region involved in binding to target antigens, and a constant ("C") region that interacts with other components of the immune system. The light chain and heavy chain variable regions (also referred herein as the light chain variable (VL) domain and heavy chain variable (VH) domain, respectively) assemble in three-dimensional space to form a variable region that binds to antigens (e.g., receptors on the surface of cells). Within each light chain or heavy chain variable region, there are three short segments referred to as complementarity determining regions ("CDR") (average length of about 3 to about 16 amino acids). The six CDRs (three from the light chain and three from the heavy chain) in the antibody variable domain fold together in three-dimensional space to form the actual antibody binding site that binds to the target antigen. The positions and lengths of the CDRs are in the Kabat numbering system; literature [Kabat et al. , Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991).], or the IMGT numbering system; it may be defined according to the literature [Lefranc, MP et al. Nucl. Acids Res. 27:209-212 (1999)], and the literature [Ruiz, M. e al. Nucl. Acids Res. 28:219-221 (2000)]. In an embodiment, if the CDR is numbered according to the Kavat numbering system, V L At approximately residuals 24-34 (L1), 50-56 (L2), and 89-97 (L3), and V HIn this case, it is approximately 31-35 (H1), 50-66 (H2), and 99-111 (H3). In an embodiment, when the CDR is numbered according to the IMGT numbering system, V L At approximately residuals 27-32 (L1), 50-52 (L2), and 89-97 (L3) and V H These are 26-33 (H1), 51-58 (H2), and 97-111 (H3). A portion of the variable region not included in the CDR is referred to as the framework ("FR"), which forms the environment for the CDR.

[0070] "Antibody fragment" as provided herein refers to a polypeptide that binds to an antigen and may comprise one or more structural domains of an antibody (e.g., light chain variable domain, heavy chain variable domain). Non-limiting examples of antibody fragments include single-domain antibodies or nanobodies, single-specific Fab2, bispecific Fab2, trispecific Fab3, monovalent IgGs, scFv, bispecific antibodies, bispecific diabodies, trispecific triabodidies, scFv-Fc, minibodies, IgNAR, V-NAR, hcIgG, VhH, or peptibodidies. "Peptibodidy" as provided herein refers to a peptide moiety attached to the Fc domain of an antibody (via a covalent or non-covalent linker). Additional non-limiting examples of antibodies known in the art include antibodies produced by cartilaginous fish or camelid animals. A general description of antibodies from camelid animals, their variable regions, and methods of their production, isolation, and use can be found in references WO97 / 49805 and WO97 / 49805, the full text of which is incorporated herein by reference for all purposes. Likewise, antibodies from cartilaginous fish, their variable regions, and methods of their production, isolation, and use can be found in WO2005 / 118629, the full text of which is incorporated herein by reference for all purposes.

[0071] The terms "CDR L1," "CDR L2," and "CDR L3" as provided herein refer to complementarity determining regions (CDRs) 1, 2, and 3 of the variable light chain (L) of the antibody. In an embodiment, the variable light chain provided herein comprises CDR L1, CDR L2, and CDR L3 in the direction from the N-terminus to the C-terminus. Likewise, the terms "CDR H1," "CDR H2," and "CDR H3" as provided herein refer to complementarity determining regions (CDRs) 1, 2, and 3 of the variable heavy chain (H) of the antibody. In an embodiment, the variable heavy chain provided herein comprises CDR H1, CDR H2, and CDR H3 in the direction from the N-terminus to the C-terminus.

[0072] The terms "FR L1," "FR L2," "FR L3," and "FR L4" as provided herein are used in accordance with their general meaning in the art and refer to framework regions (FR) 1, 2, 3, and 4 of the variable light chain (L) of the antibody. In an embodiment, the variable light chain provided herein comprises FR L1, FR L2, FR L3, and FR L4 in the direction from the N-terminus to the C-terminus. Likewise, the terms "FR H1," "FR H2," "FR H3," and "FR H4" as provided herein are used in accordance with their general meaning in the art and refer to framework regions (FR) 1, 2, 3, and 4 of the variable heavy chain (H) of the antibody. In an embodiment, the variable heavy chain provided herein comprises FR H1, FR H2, FR H3, and FR H4 in the direction from the N-terminus to the C-terminus.

[0073] An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer consists of two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily associated with antigen recognition. The terms variable light chain (VL), variable light chain (VL) domain or light chain variable region and variable heavy chain (VH), variable heavy chain (VH) domain or heavy chain variable region refer to these light chain and heavy chain regions, respectively. The terms variable light chain (VL), variable light chain (VL) domain, and light chain variable region as referred herein may be used interchangeably. The terms variable heavy chain (VH), variable heavy chain (VH) domain, and heavy chain variable region as referred herein may be used interchangeably. The Fc (i.e., fragment crystallizable region) is the "base" or "tail" of the immunoglobulin and typically consists of two heavy chains that provide two or three constant domains, depending on the class of antibody. By binding to a specific protein, the Fc region ensures that each antibody elicits an appropriate immune response against a given antigen. The Fc region also binds to various cell receptors, such as Fc receptors, and other immune molecules, such as complement proteins.

[0074] The term "antibody" is used in accordance with its generally known meaning in the art. Antibodies exist, for example, as intact immunoglobulins or as numerous well-characterized fragments produced by degradation by various peptidases. Accordingly, for example, pepsin degrades the antibody under the disulfide linkage at the hinge region to produce the F(ab)'2, Fab dimer, which itself is V by the disulfide linkage H -C H1It is a light chain connected to. F(ab)'2 can be reduced under mild conditions to cleave the disulfide link at the hinge region, thereby converting the F(ab)'2 dimer into a Fab' monomer. The Fab' monomer is essentially a Fab having part of the hinge region (see [Fundamental Immunology (Paul ed., 3d ed. 1993], the full text of which is incorporated herein for all purposes). Various antibody fragments are defined in terms of the degradation of an intact antibody, and those skilled in the art will understand that such fragments may be newly synthesized chemically or using recombinant DNA methods. Accordingly, the term antibody as used herein also includes antibody fragments produced by modification of a whole antibody, those newly synthesized using recombinant DNA methods (e.g., single-stranded Fv), or those identified using a phage display library (e.g., [McCafferty et al. , Nature [See 348:552-554 (1990)], the full text of which is incorporated herein for all purposes). The term “antibody” as used herein further comprises antibody fragments, such as single-domain antibodies. Accordingly, in the embodiments, the antibody comprises a single monomeric variable antibody domain. Accordingly, in the embodiments, the antibody comprises a variable light chain (VL) domain or a variable heavy chain (VH) domain. In the embodiments, the antibody is a variable light chain (VL) domain or a variable heavy chain (VH) domain.

[0075] For the production of monoclonal or polyclonal antibodies, any technique known in the art may be used (e.g., the literature [Kohler & Milstein, Nature 256:495-497 (1975)]; literature[Kozbor et al. , Immunology Today 4:72 (1983)]; Literature[Cole et al ., pp. 77-96 in Monoclonal Antibodies and Cancer Therapy[See (1985)], the full text of which is incorporated herein for all purposes). "Monoclonal" antibodies (mAbs) refer to antibodies derived from a single clone. Techniques for the production of single-chain antibodies (U.S. Patent No. 4,946,778, the full text of which is incorporated herein for all purposes) may be adopted to produce antibodies against the polypeptide of the present invention. Additionally, transgenic mice, or other organisms such as mammals, may be used to express humanized antibodies. Alternatively, phage display techniques may be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (e.g., the literature [McCafferty et al. , Nature 348:552-554 (1990)]; Literature[Marks et al. , Biotechnology See 10:779-783 (1992)], the full text of which is incorporated herein for all purposes).

[0076] A single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy chain (VH) and light chain (VL) of an immunoglobulin, typically linked to a short linker peptide of 10 to about 25 amino acids. The linker can usually be glycine-rich for flexibility, as well as serine or threonine-rich for solubility. The linker can connect the N-terminus of the VH to the C-terminus of the VL, or vice versa.

[0077] Another example of an antibody fragment according to the initiation is the Fab' fragment. The Fab' fragment differs from the Fab fragment in the presence of one or more additional residues containing cysteine ​​from the antibody hinge region at the C-terminus of the CH1 domain. The F(ab')2 antibody fragment is a pair of Fab' fragments linked by cysteine ​​residues at the hinge region. Other chemical couplings of antibody fragments are also known.

[0078] The "Fv" fragment contains a complete antigen-recognition and binding site consisting of a dimer of one heavy chain and one light chain variable domain that are closely non-covalently associated. In this structure, three CDRs of each variable domain interact to define the antigen-binding site on the surface of the VH-VL dimer. Collectively, six CDRs confer antigen-binding specificity to the antibody.

[0079] The antibody fragment can also form a tandem Fd segment, which forms a pair of antigen-binding regions including a pair of tandem Fd segments (VH-CH1-VH-CH1). These "linear antibodies" can be bispecific or monospecific, for example, as described in the literature [Zapata et al. 1995, Protein Eng. 8(10):1057-1062].

[0080] The epitope of an mAb is the region of the antigen to which the mAb binds. Two antibodies bind to the same or overlapping epitopes when they each competitively inhibit (block) the binding of the other to an antigen. That is, one antibody in excess of 1x, 5x, 10x, 20x, or 100x inhibits the binding of the other by at least 30%, but preferably by 50%, 75%, 90%, or even 99%, as measured in a competitive binding assay (e.g., the literature [Junghans et al [See Cancer Res. 50:1495, 1990, the full text of which is incorporated herein for all purposes). Alternatively, if essentially all amino acid mutations in an antigen that reduce or eradicate the binding of one antibody reduce or eradicate the binding of another, the two antibodies have the same epitope. If some amino acid mutations that reduce or eradicate the binding of one antibody reduce or eradicate the binding of another, the two antibodies have overlapping epitopes.

[0081] For the preparation of a suitable antibody of the invention for use according to the invention, e.g., a recombinant, monoclonal, or polyclonal antibody, numerous techniques known in the art may be used (see, for example, [Kohler & Milstein, Nature 256:495-497 (1975)]; [Kozbor et al., Immunology Today 4: 72 (1983)]; [Cole et al., pp. 77-96 in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc. (1985)]; [Coligan, Current Protocols in Immunology (1991)]; [Harlow & Lane, Antibodies, A Laboratory Manual (1988)]; and [Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986)], the full text of which is incorporated herein for all purposes). Genes encoding the heavy and light chains of interest may be cloned from cells, for example, genes encoding monoclonal antibodies may be cloned from hybridomas and used to produce recombinant monoclonal antibodies. Gene libraries encoding the heavy and light chains of monoclonal antibodies may also be prepared from hybridomas or plasma cells. Random combinations of heavy and light chain gene products produce a large population of antibodies with different antigen specificities (see, for example, Kuby, Immunology (3rd ed. 1997) [the full text of which is incorporated herein for all purposes]). Techniques for producing monochain antibodies or recombinant antibodies (U.S. Patent No. 4,946,778, U.S. Patent No. 4,816,567, the full text of which is incorporated herein for all purposes) may be adopted to produce antibodies against the polypeptide of the present invention.Additionally, transgenic mice, or other organisms such as other mammals, may be used for humanization or to express human antibodies (e.g., U.S. Patents No. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, [Marks et al., Bio / Technology 10:779-783 (1992)]; [Lonberg et al., Nature 368:856-859 (1994)]; [Morrison, Nature 368:812-13 (1994)]; [Fishwild et al., Nature Biotechnology 14:845-51 (1996)]; [Neuberger, Nature Biotechnology 14:826 (1996)]; and see [Lonberg & Huszar, Intern. Rev. Immunol. 13:65-93 (1995)], the full text of which is incorporated herein for all purposes). Alternatively, phage display technology may be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (e.g., see [McCafferty et al., Nature 348:552-554 (1990)]; see [Marks et al., Biotechnology 10:779-783 (1992)], the full text of which is incorporated herein for all purposes). Antibodies can also be produced bispecifically, that is, capable of recognizing two different antigens (see, e.g., WO 93 / 08829, reference [Traunecker et al., EMBO J. 10:3655-3659 (1991)]; and reference [Suresh et al., Methods in Enzymology 121:210 (1986)], the full text of which is incorporated herein for all purposes).The antibody may also be a heterozygous, for example, two covalently bound antibodies, or an immunotoxin (e.g., U.S. Patent No. 4,676,980, WO 91 / 00360; WO 92 / 200373; and EP 03089, the full text of which is incorporated herein for all purposes).

[0082] Methods for humanizing or primating non-human antibodies are well known in the art (see, e.g., U.S. Patents No. 4,816,567; 5,530,101; 5,859,205; 5,585,089; 5,693,761; 5,693,762; 5,777,085; 6,180,370; 6,210,671; and 6,329,511; WO 87 / 02671; EP Application 0173494; [Jones et al. (1986) Nature 321:522]; and [Verhoyen et al. (1988) Science 239:1534], the full text of which is incorporated herein for all purposes). Humanized antibodies are further described, for example, in the literature [Winter and Milstein (1991) Nature 349:293], the full text of which is incorporated herein for all purposes. Generally, humanized antibodies have one or more amino acid residues from which they are injected from a non-human source. These non-human amino acid residues are often referred to as import residues, which are typically obtained from an import variable domain. Humanization can essentially be carried out according to the methods of Winter and his associates (e.g., [Morrison et al., PNAS USA, 81:6851-6855 (1984)], [Jones et al., Nature 321:522-525 (1986)]; [Riechmann et al., Nature 332:323-327 (1988)]; [Morrison and Oi, Adv. Immunol., 44:65-92 (1988)], [Verhoeyen et al., Science 239:1534-1536 (1988)] and [Presta, Curr. Op. Struct. Biol. 2:593-596 (1992)], [Padlan, Molec. Immun., 28:489-498 (1991)]; literature[Padlan, Molec. Immun.[See , 31(3):169-217 (1994)], the full text of which is incorporated herein for all purposes), which is achieved by substituting the corresponding sequence of a human antibody with a rodent CDR or CDR sequence. Thus, such humanized antibodies are chimeric antibodies (U.S. Patent No. 4,816,567, the full text of which is incorporated herein for all purposes), wherein a significantly smaller portion than the intact human variable domain is substituted with a corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted with residues from a similar region of a rodent antibody. For example, a polynucleotide comprising a first sequence encoding a humanized immunoglobulin framework region and a second set of sequences encoding a desired immunoglobulin complementarity determining region can be produced synthetically or by combining appropriate cDNA and genomic DNA segments. The human constant region DNA sequence can be isolated from various human cells according to well-known procedures.

[0083] A “chimeric antibody” is (a) an antibody molecule in which an invariant region or a part thereof is modified, replaced, or exchanged so that the antigen-binding site (variable region) is connected to an invariant region of a different or modified class, effector function, and / or species, or a completely different molecule, e.g., an enzyme, toxin, hormone, growth factor, drug, etc., which confers new characteristics to the chimeric antibody; or (b) an antibody molecule in which an invariant region or a part thereof is modified, replaced, or exchanged to have a variable region having different or modified antigen specificity. An antibody preferred for use in accordance with the invention comprises a humanized and / or chimeric monoclonal antibody.

[0084] When referring to proteins or peptides, the phrases "specifically (or selectively) bind" or "specifically (or selectively) immunoreactive to" often relate to binding reactions that determine the presence of a protein in a heterogeneous population of proteins and other biological agents. Accordingly, under specified immunoassay conditions, a specific antibody binds to a specific protein at least twice the background level, and more typically more than 10 to 100 times the background level. Under these conditions, specific binding to an antibody requires an antibody selected for its specificity to a specific protein. For example, polyclonal antibodies may be selected to obtain only a subgroup of antibodies that are specifically immunoreactive to a selected antigen rather than to other proteins. This selection can be achieved by removing antibodies that cross-react with other molecules. Various immunoassay formats may be used to select antibodies that are specifically immunoreactive to a specific protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies that are specifically immunoreactive to proteins (see, for example, the literature [Harlow & Lane, Using Antibodies, A Laboratory Manual (1998)], the full text of which describes the formats and conditions of immunoassays that may be used to determine specific immunoreactivity for all purposes, incorporated herein).

[0085] "Ligand" refers to a preparation capable of binding to a receptor or antibody, antibody variant, antibody region or fragment thereof, e.g., polypeptide or other molecule.

[0086] Techniques for conjugating therapeutic agents to antibodies are well known (e.g., see the following literature: [Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985)]; [Hellstrom et al., "Antibodies For Drug Delivery" in Controlled Drug Delivery (2 nd [Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review" in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985)]; and [Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev., 62:119-58 (1982)], the full text of which is incorporated herein for all purposes). As used herein, the term "antibody-drug conjugate" or "ADC" refers to a therapeutic agent that is conjugated to or otherwise covalently bonded to an antibody.

[0087] The terms "WISP1 protein" or "WISP1" as used herein refer to Cell Communication Network Factor 4, CCN family member 4, It comprises any one of the Wnt-1-induced secretory protein, CCN4, or WISP1-induced signaling pathway protein 1 (WISP1), which is also known as a variant or homologue thereof that retains WISP1 activity (e.g., within 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% activity compared to WISP1). In some embodiments, the variant or homologue has at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity with respect to the whole sequence or a portion of the sequence (e.g., a portion of 50, 100, 150, or 200 consecutive amino acids) compared to the naturally occurring WISP1 protein. In an embodiment, the WISP1 protein is a mammalian WISP1 protein. In an embodiment, the WISP1 protein is a human WISP1 protein. In an embodiment, the WISP1 protein is substantially identical to the protein identified by SEQ ID NO. 15. In an embodiment, the WISP1 protein is substantially identical to the protein identified by SEQ ID NO. 16. In an embodiment, the WISP1 protein is substantially identical to the protein identified by SEQ ID NO. 17. In an embodiment, the WISP1 protein is substantially identical to the protein identified by SEQ ID NO. 19. In an embodiment, the WISP1 protein is substantially identical to the protein identified by SEQ ID NO. 20. In an embodiment, the WISP1 protein is substantially identical to the protein identified by UniProt reference ID NO. 095388, or is a variant or homologue having substantial identity therewith. In an embodiment, the WISP1 protein comprises any one of domains 1–4 or a combination thereof. Accordingly, in an embodiment, the WISP1 protein comprises domain 1, domain 2, domain 3, or domain 4 of the WISP1 protein. In an embodiment, domain 1 comprises the sequence of SEQ ID NO. 25. In an embodiment, the WISP1 protein comprises the sequence of sequence number 25.In an embodiment, domain 2 comprises the sequence of SEQ ID NO. 26. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 26. In an embodiment, domain 3 comprises the sequence of SEQ ID NO. 27. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 27. In an embodiment, domain 4 comprises the sequence of SEQ ID NO. 28. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 28.

[0088] For the specific proteins described herein, the named protein comprises any one of the naturally occurring form of the protein, a variant, or a homologue that retains protein activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% activity compared to the natural protein). In some embodiments, the variant or homologue has at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity with respect to the whole sequence or a portion of the sequence (e.g., a 50, 100, 150, or 200 consecutive amino acid portion) compared to the naturally occurring form. In other embodiments, the protein is a protein identified by its NCBI sequence criteria. In other embodiments, the protein is a protein, its homologue, or a functional fragment identified by its NCBI sequence criteria.

[0089] The term "gene" refers to a segment of DNA involved in protein synthesis; it includes regions preceding and following the coding region (leaders and trailers), as well as sequences interposed between individual coding segments (exons) (introns). Leaders, trailers, and introns contain regulatory elements necessary for the transcription and translation processes of the gene. Furthermore, a "protein gene product" is a protein expressed from a specific gene.

[0090] The terms "plasmid," "vector," or "expression vector" refer to nucleic acid molecules that code for genes and / or regulatory elements necessary for gene expression. Gene expression from a plasmid can occur in a cis or trans form. When a gene is expressed in a cis form, the gene and regulatory element are encoded by the same plasmid. Expression in a trans form involves cases where the gene and regulatory element are encoded by separate plasmids.

[0091] The terms “transfusion,” “transduction,” “transfused,” or “transduced” may be used interchangeably and are defined as the process of introducing nucleic acid molecules or proteins into cells. Nucleic acids are introduced into cells using non-viral or virus-based methods. Nucleic acid molecules may be gene sequences encoding a complete protein or a functional part thereof. Non-viral methods of transfusion include any suitable transfusion method that does not use viral DNA or viral particles as a delivery system for introducing nucleic acid molecules into cells. Exemplary non-viral transfusion methods include calcium phosphate transfusion, liposomal transfusion, nucleofection, sonoporation, transfusion via heat shock, magnetifection, and electroporation. In some embodiments, nucleic acid molecules are introduced into cells using electroporation according to standard procedures well known in the art. For virus-based methods of transduction, any useful viral vector may be used in the methods described herein. Examples of viral vectors include, but are not limited to, retroviruses, adenoviruses, lentiviruses, and adeno-associated virus vectors. In some embodiments, nucleic acid molecules are introduced into cells using retrovirus vectors according to standard procedures well known in the art. The terms "transduction" or "transduction" also refer to the introduction of a protein from the external environment into a cell. Typically, the transduction or transduction of a protein relies on the attachment of a peptide or protein capable of passing through the cell membrane to the protein of interest. For example, see the following: Literature [Ford et al. (2001) Gene Therapy 8:1-4 and Prochiantz (2007) Nat. Methods 4:119-20], the full text of which is incorporated herein for all purposes.

[0092] A “label” or “detectable moiety” is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. For example, useful labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., commonly used in ELISA), biotin, digoxigenin, or hapten, and other substances that can be made detectable by incorporating the radiolabel into proteins or, for example, peptides or antibodies that are specifically reactive to a target peptide. Any suitable method known in the art for conjugating antibodies to labels may be used, for example, by using the method described in the literature in which the full text thereof is incorporated herein for all purposes [Hermanson, Bioconjugate Techniques 1996, Academic Press, Inc., San Diego.].

[0093] "Contact" is used in its literal, general sense and refers to a process in which at least two distinct species (e.g., antibody and antigen) are brought into sufficient proximity to react, interact, or come into physical contact. However, it should be understood that the resulting reaction product may be produced directly from the reaction between the added reagents, or from an intermediate from one or more added reagents that may be produced within the reaction mixture.

[0094] The term “contact” may include causing two species to react, interact, or come into physical contact, wherein the two species may be, for example, a pharmaceutical composition and a cell as provided herein. In an embodiment, contact includes, for example, causing an antibody as described herein to interact with a cell. In an embodiment, contact includes, for example, causing an antibody as described herein to physically interact with a WISP1 protein or a fragment thereof.

[0095] As used herein, “cell” refers to a cell that performs sufficient metabolic or other functions to preserve or replicate its genomic DNA. Cells may be identified by methods well known in the art, including, for example, the presence of an intact membrane, staining by a specific dye, the ability to produce offspring, or, in the case of germ cells, the ability to combine with a second germ cell to produce viable offspring. Cells may include prokaryotic cells and eukaryotic cells. Prokaryotic cells include, but are not limited to, bacteria. Eukaryotic cells include, but are not limited to, enzyme cells and cells derived from plants and animals, e.g., mammals, insects (e.g., spodoptera), and human cells.

[0096] The term "recombinant" indicates, when used to refer to, for example, cells, nucleic acids, proteins, or vectors, that the cells, nucleic acids, proteins, or vectors have been modified by the introduction of heterologous nucleic acids or proteins or by alteration of the original nucleic acids or proteins, or that the cells originated from such modified cells. Accordingly, for example, recombinant cells express genes not found in the original (non-recombinant) form of the cell, or express original genes that are otherwise abnormally expressed, under-expressed, or not expressed at all. Transgenic cells and plants generally express heterologous genes or coding sequences through recombinant methods.

[0097] The term "isolated," when applied to nucleic acids or proteins, means that the nucleic acid or protein is essentially free of other cellular components that bind to it in its natural state. This may be, for example, in a homogeneous state, dried, or in an aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high-performance liquid chromatography. Proteins, which are the species predominantly present in the formulation, are significantly purified.

[0098] When the term "heterogeneous" is used to refer to a portion of nucleic acid, it indicates that the nucleic acid comprises two or more subsequences that are not found in nature in an identical relationship to one another. For example, nucleic acids are typically produced by recombination to have two or more sequences from unrelated genes arranged to create a new functional nucleic acid, such as a promoter from one source and a coding region from another source. Similarly, a heterogeneous protein indicates that a protein comprises two or more subsequences that are not found in nature in an identical relationship to one another (e.g., a fusion protein).

[0099] The term "exogenous" relates to molecules or substances (e.g., compounds, nucleic acids, or proteins) originating from outside a given cell or organism. For example, an "exogenous promoter" as referred to herein is a promoter that does not originate from the cell or organism expressing it. On the other hand, the terms "endogenous" or "endogenous promoter" refer to molecules or substances that are inherent in a given cell or organism or originate therefrom.

[0100] As defined herein, terms such as “inhibition,” “inhibits,” and “inhibiting,” when referring to protein activity, mean having a negative effect on the function of a protein (e.g., reducing it). In some embodiments, inhibition relates to the reduction of a disease or symptoms of a disease (e.g., fibrosis (e.g., hepatic fibrosis, pulmonary fibrosis, etc.)). Accordingly, inhibition includes blocking a stimulus at least partially, partially, or wholly; reducing, preventing, or delaying activation; or inactivating, desensitizing, or downregulating signal transduction or enzymatic activity or the amount of a protein (e.g., WISP1 protein). Similarly, “inhibitor” refers, for example, , It is a compound or protein that inhibits a receptor or other protein by binding to, partially or completely blocking, reducing, preventing, delaying, inactivating, desensitizing, or downregulating the activity (e.g., receptor activity or protein activity).

[0101] As defined herein, terms such as “inhibition,” “inhibits,” and “inhibiting” when referring to protein-inhibitor interactions mean having a negative effect (e.g., reducing) on ​​the activity or function of a protein (e.g., WISP1 protein) compared to the activity or function of the protein in the absence of an inhibitor. In the embodiments, inhibition means having a negative effect (e.g., reducing) on ​​the concentration or level of WISP1 compared to the concentration or level of the protein in the absence of an inhibitor. In the embodiments, inhibition relates to a reduction in a disease or symptoms of a disease (e.g., fibrosis). In the embodiments, inhibition relates to a reduction in the activity of WISP1. Accordingly, inhibition includes blocking a stimulus at least partially, partially or wholly, reducing, preventing, or delaying activation, or inactivating, desensitizing, or downregulating signal transduction or enzymatic activity or the amount of WISP1. In the embodiments, inhibition relates to a reduction in the activity of WISP1 resulting from a direct interaction (e.g., the inhibitor is bound to WISP1). In an embodiment, inhibition is associated with a reduction in the activity of WISP1 from an indirect interaction (e.g., the inhibitor binds to a protein that activates WISP1, thereby preventing the activation of the target protein).

[0102] Accordingly, the terms “inhibitor,” “inhibitory substance,” “antagonist,” or “down-regulator” refer interchangeably to a substance capable of detectably reducing the expression or activity of a specific gene or protein (e.g., WISP1 protein). An antagonist may reduce WISP1 expression or activity by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the control group in the absence of the antagonist. In certain cases, WISP1 expression or activity is 1.5 times, 2 times, 3 times, 4 times, 5 times, or 10 times less than the expression or activity in the absence of the antagonist.

[0103] The term “expression” includes, but is not limited to, any step associated with the production of a polypeptide, including transcription, post-transcriptional modification, translation, post-translational modification, and secretion. Expression may be detected using conventional techniques for detecting proteins (e.g., ELISA, Western blotting, flow cytometry, immunofluorescence, immunohistochemistry, etc.).

[0104] “Biological sample” or “sample” refers to material obtained from or derived from a subject or patient. Biological samples include sections of tissue, such as biopsy and autopsy samples taken for histological purposes, and frozen sections. These samples include body fluids, such as blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, etc.), sputum, tissues, cultured cells (e.g., primary cultures, grafts, and transformed cells), feces, urine, immune cells, hematopoietic cells, fibroblasts, macrophages, T cells, etc. Biological samples are typically obtained from eukaryotic organisms, such as mammals, such as primates, such as chimpanzees or humans; cattle; dogs; cats; rodents, such as guinea pigs, rats, mice; rabbits; or birds; reptiles; or fish. In an embodiment, the biological sample is bronchoalveolar lavage fluid (BALF). In an embodiment, the biological sample is plasma. In an embodiment, the biological sample is tissue derived from the subject’s tissue, for example, lungs, liver, or heart.

[0105] A "control group" or "standard control group" relates to a reference for comparison with a test sample, measurement, or value, and to a sample, measurement, or value that serves as a generally known standard. For example, a test sample may be taken from a patient suspected of having a specific disease (e.g., fibrosis) and compared to a known normal (unaffected) individual (e.g., a standard control subject). In an embodiment, the control group is a biological sample taken from an individual without fibrosis. A standard control group may also represent an average measurement or value obtained from a similar population of individuals without the specific disease (e.g., a standard control subject), for example, healthy individuals with similar medical history, same age, weight, etc. Standard control values ​​may also be obtained from the same individual, for example, from a sample taken early from the patient prior to the onset of the disease. For example, a control group may be designed to compare therapeutic benefits based on pharmacological data (e.g., half-life) or therapeutic measurements (e.g., comparison of side effects). A control group is also useful for determining the significance of data. For example, in the embodiments, the control group may be a test sample that does not contain the anti-WISP1 antibody as provided herein. For example, if the value for a given parameter varies widely in the control group, the change in the test sample will not be considered significant. Those skilled in the art will recognize that the standard control group may be designed for the evaluation of any number of parameters (e.g., RNA levels, protein levels, specific cell types, specific body fluids, specific tissues, etc.).

[0106] Those skilled in the art will understand that a standard control group is most appropriate in a given situation and that data can be analyzed based on comparisons to the standard control group values. Standard control groups are also useful for determining the significance of data (e.g., statistical significance). For example, if the value of a given parameter varies widely in the standard control group, the change in the test sample will not be considered significant.

[0107] “Patient” or “object requiring the same” refers to a living organism suffering from or susceptible to a disease (e.g., fibrosis) or pathological condition that can be treated by the administration of a composition or pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cattle, deer, and other non-mammalians. In some embodiments, the patient is a human.

[0108] The terms “disease” or “pathological condition” relate to the condition or health status of a patient or subject that may be treated by the compound or method provided herein. The disease may be fibrosis. Fibrosis is a pathological condition that may be characterized by the abnormal accumulation of collagen matrix following injury or inflammation that alters the structure and function of various tissues. In embodiments, fibrosis is in the kidney, liver, lungs, heart, bone or bone marrow, or skin.

[0109] In the context of a substance or substance activity or function associated with a disease, the term “associated” or “related thereto” (e.g., protein-associated disease (e.g., fibrosis associated with WISP1 activity)) means that the disease (e.g., fibrosis) is caused (wholly or partially) by the substance or substance activity or function, or that the symptoms of the disease are caused (wholly or partially) by it. As used herein, if what is described as associated with the disease is a causative substance, it may be a target for the treatment of the disease. For example, fibrosis associated with WISP1 activity or function or WISP1-associated disease (e.g., fibrosis (e.g., hepatic fibrosis, pulmonary fibrosis, etc.)) may be treated with a WISP1 modulator or a WISP1 inhibitor if increased WISP1 activity or function (e.g., signaling pathway activity) causes the disease (e.g., fibrosis). For example, fibrotic disorders associated with WISP1 activity can be treated with WISP1 modulators or WISP1 inhibitors when increased WISP1 activity or function (e.g., signaling pathway activity) causes the disease.

[0110] The term "signaling pathway" as used herein relates to a series of interactions between a cell and optionally extracellular components (e.g., proteins, nucleic acids, small molecules, ions, lipids), which transmit a change in one component to one or more other components, which can eventually transmit a change to additional components, which then propagate optionally to other signaling pathway components.

[0111] The term "abnormal" as used herein relates to something different from normal. When used to describe enzyme activity, abnormal relates to activity that is greater or less than the mean of a normal control or a normal, unaffected control sample. Abnormal activity may relate to a disease-causing amount of activity, wherein returning abnormal activity to a normal or non-disease-related amount (by using the methods described herein) results in a reduction of the disease or one or more disease symptoms.

[0112] As referred to herein, a “therapeutic agent” is a composition useful for treating or preventing fibrosis (e.g., hepatic fibrosis, pulmonary fibrosis, etc.). In an embodiment, the therapeutic agent is an anti-fibrotic agent. For example, an anti-fibrotic agent may refer to a composition (e.g., a compound, drug, antagonist, inhibitor, modifier) ​​having the ability to inhibit or slow the rate of fibrosis or scar formation. In an embodiment, the anti-fibrotic agent is a preparation approved by the FDA or a similar regulatory agency in a country other than the United States for treating fibrosis.

[0113] As used herein, “treating” or “treating” a condition, disease, or disorder or symptoms associated with a condition, disease, or disorder relates to a method for obtaining a favorable or desirable outcome, including clinical outcomes. Favorable or desirable clinical outcomes may include, without limitation, alleviation or improvement of one or more symptoms or conditions; reduction in the degree of a condition, disorder, or disease; stabilization of the condition, disorder, or disease state; prevention of the onset of a condition, disorder, or disease; prevention of the spread of a condition, disorder, or disease; delay or slowing the progression of a condition, disorder, or disease; delay or slowing the onset of a condition, disorder, or disease; alleviation or reduction of the condition, disorder, or disease state; and remission, whether partial or total. “Treating” may also mean an extended lifespan of the subject compared to what would be expected without treatment. “Treating” may also mean suppressing the progression of a condition, disorder, or disease, or temporarily delaying the progression of a condition, disorder, or disease, and in some cases, this entails permanently stopping the progression of a condition, disorder, or disease. As used herein, the terms treatment, to treat, or treating relate to a method of reducing the effect of one or more symptoms of a disease or condition characterized by the expression of protease or symptoms of a disease or condition characterized by the expression of protease. Accordingly, in the method disclosed, treatment may relate to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of an established disease, condition, or symptoms of a disease or condition. For example, a method of treating a disease is considered to be treatment if there is a 10% reduction in one or more symptoms of the disease in the subject compared to a control group.Accordingly, reduction may be a percentage reduction of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage between 10% and 100% compared to the original or control level. It should be understood that treatment does not necessarily mean curing or completely eliminating the disease, condition, or symptoms of the disease or condition. Furthermore, as used herein, references to reduction, decrease, or inhibition include a change of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more compared to the control level, and these terms may include, but do not necessarily include, complete elimination.

[0114] The terms “dosage” and “administration amount” are used interchangeably herein. Dosage refers to the amount of active ingredient given to an individual at each administration. Dosage will vary depending on a number of factors, including the normal dose range for a given regimen, frequency of administration; size and tolerance of the individual; severity of the condition; risk of adverse effects; and route of administration. Those skilled in the art will recognize that dosage may vary depending on the aforementioned factors or based on the course of treatment. The term “formulation” refers to a specific form of the drug or pharmaceutical composition and depends on the route of administration. For example, a formulation may be a liquid form for nebulization, e.g., for inhalation; a tablet or liquid for oral delivery, e.g.; or a saline solution for injection, e.g.

[0115] As used herein, “therapeutic effective dose” means any amount that produces the effect of administration (e.g., treatment or prevention of a disease). As used herein, “therapeutic effective dose” means a specific amount of a substance (e.g., antibodies disclosed herein) administered to a subject in a single dose or over a specific period. “Therapeutic effective dose” may comprise one or more “therapeutic effective dose(s)” necessary to achieve the desired therapeutic effect. The precise dose and formulation will depend on the therapeutic purpose and will be determined by a person skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999)]; Remington, The Science and Practice of Pharmacy, 20th Edition, Gennaro, Editor (2003)]; and Pickar, Dosage Calculations (1999)—the full text of which is incorporated herein for all purposes). For example, for a given parameter, the therapeutically effective dose will represent an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy may also be expressed as an increase or decrease of "number of times." For example, the therapeutically effective dose may have an effect of at least 1.2 times, 1.5 times, 2 times, or 5 times or more compared to a standard control group. The therapeutically effective dose or amount may alleviate one or more symptoms of the disease. The therapeutically effective dose or amount may prevent or delay the onset of the disease or one or more symptoms of the disease when treating a person at risk of developing the disease as a result of its administration.

[0116] As used herein, the term “administering” means oral administration to a subject, administration as a suppository, local contact, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal, or subcutaneous administration, or implantation of a sustained-release device, e.g., a mini-osmotic pump. Administration is by any route including parenteral and transmucosal. Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other delivery methods include, but are not limited to, the use of liposomal preparations, intravenous infusion, transdermal patches, etc. . "Co-administered" means that the compositions described herein are administered simultaneously, immediately before, or immediately after the administration of one or more additional therapies, e.g., antifibrotic agents. The compounds of the invention may be administered to a patient alone or co-administered. Co-administration means administering the compounds individually or in combination (with more than one compound) simultaneously or sequentially. Accordingly, the formulations may also be combined with other active substances if desired (e.g., to reduce metabolic degradation). The compositions of the invention are delivered via transdermal or topical routes and may be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.

[0117] As used herein, the term "pharmaceutically acceptable" is used synonymously with "physiologically acceptable" and "pharmacologically acceptable." Pharmaceutical compositions will generally contain buffers and preservatives for storage and may contain carriers and buffers for appropriate delivery depending on the route of administration.

[0118] "Pharmaceuticalally acceptable excipients" and "pharmaceuticalally acceptable carriers" refer to substances that support the administration of the active agent and the absorption of the target, and may be included in the composition of the present invention without causing serious toxicological side effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline, lactate Ringer's solution, general sucrose, general glucose, binders, fillers, disintegrants, lubricants, coating agents, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, and dyes, etc. These preparations are sterile and, if desired, may be mixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, osmotic pressure-regulating salts, buffers, coloring agents and / or flavoring substances, which do not react harmfully with the compounds of the present invention. Those skilled in the art will recognize that other pharmaceutical excipients are useful for the present invention.

[0119] The term “pharmaceutically acceptable salt” refers to salts derived from various organic and inorganic counterions well known in the art, merely examples including sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc.; and, where the molecule contains a basic functional group, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, malate, oxalate, etc.

[0120] The term “preparation” is intended to include a preparation of an active compound having an encapsulating material as a carrier providing a capsule, wherein an active ingredient with or without another carrier is surrounded by the carrier and is thus combined therewith. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges may be used as solid formulations suitable for oral administration.

[0121] Pharmaceutical formulations are optionally present in unit formulations. In this form, the formulation is subdivided into unit doses containing a suitable amount of the active ingredient. The unit formulation may be a packaged formulation, and the package contains individual amounts of formulations, packaged tablets, capsules, and powders in vials or ampoules. Additionally, the unit formulation may be a capsule, tablet, case, or lozenge itself, or it may be any one of these in an appropriate number of packaged forms. The unit formulation may be a frozen dispersion.

[0122] The embodiments and modes of embodiment described herein are for illustrative purposes only, and various modifications or variations related thereto are to be proposed to those skilled in the art and are understood to be included within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

[0123] Anti-WISP1 antibody

[0124] Above all, antibodies (e.g., humanized antibodies, monoclonal antibodies) and antibody fragments (e.g., scFv) that bind to WNT1-induced signaling pathway protein 1 (WISP1) with high efficiency and specificity are provided herein. The antibodies and antibody compositions provided herein contain novel light chain and heavy chain domains CDR and have been confirmed to bind to domain 2 of the WISP1 protein. The antibodies described herein, including embodiments thereof, are useful for binding to WISP1 with high efficiency and affinity and inhibit WISP1 activity in cells expressing WISP1. The antibodies provided herein, including embodiments thereof, may be used for diagnostic and therapeutic purposes in WISP1-related fibrotic disorders. The antibodies provided herein, including embodiments thereof, have the ability to inhibit collagen formation and, accordingly, can inhibit or delay fibrosis (e.g., hepatic fibrosis, pulmonary fibrosis, etc.).

[0125] The exemplary anti-WISP1 antibodies provided herein are referred to by names (e.g., K4, K4 antibody).

[0126] In one embodiment, an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody is provided, comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and the light chain variable domain comprises CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0127] As described above, the “light chain variable (VL) domain” as provided herein refers to a variable region of the light chain of an antibody, its antibody variant, or fragment. Similarly, the “heavy chain variable (VH) domain” as provided herein refers to a variable region of the heavy chain of an antibody, its antibody variant, or fragment. The light chain variable domain and the heavy chain variable domain together form a paratope, which binds to an antigen (epitope). The paratope or antigen-binding site is formed at the N-terminus of the antibody, its antibody variant, or fragment. In an embodiment, the light chain variable (VL) domain comprises CDR L1, CDR L2, CDR L3 and FR L1, FR L2, FR L3, and FR L4 (framework region) of the antibody light chain. In an embodiment, the heavy chain variable (VH) domain comprises CDR H1, CDR H2, CDR H3 and FR H1, FR H2, FR H3, and FR H4 (framework region) of the antibody heavy chain. In an embodiment, the light chain variable (VL) domain and the light chain constant (CL) domain form part of the antibody light chain. In an embodiment, the heavy chain variable (VH) domain and the heavy chain constant (CH1) domain form part of the antibody heavy chain. In an embodiment, the heavy chain variable (VH) domain and one or more heavy chain constant (CH1, CH2, or CH3) domains form part of the antibody heavy chain. Accordingly, in an embodiment, the light chain variable (VL) domain forms part of the antibody. In an embodiment, the heavy chain variable (VH) domain forms part of the antibody. In an embodiment, the light chain variable (VL) domain forms part of the therapeutic antibody. In an embodiment, the heavy chain variable (VH) domain forms part of the therapeutic antibody. In an embodiment, the light chain variable (VL) domain forms part of the human antibody. In an embodiment, the heavy chain variable (VH) domain forms part of the human antibody. In an embodiment, the light chain variable (VL) domain forms part of the humanized antibody. In an embodiment, the heavy chain variable (VH) domain forms part of the humanized antibody.In an embodiment, the light chain variable (VL) domain forms part of a chimeric antibody. In an embodiment, the heavy chain variable (VH) domain forms part of a chimeric antibody. In an embodiment, the light chain variable (VL) domain forms part of an antibody fragment. In an embodiment, the heavy chain variable (VH) domain forms part of an antibody fragment. In an embodiment, the light chain variable (VL) domain forms part of an antibody variant. In an embodiment, the heavy chain variable (VH) domain forms part of an antibody variant. In an embodiment, the light chain variable (VL) domain forms part of Fab. In an embodiment, the heavy chain variable (VH) domain forms part of Fab. In an embodiment, the light chain variable (VL) domain forms part of scFv. In an embodiment, the heavy chain variable (VH) domain forms part of scFv.

[0128] In an embodiment, the heavy chain variable domain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 90% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 91% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 92% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 93% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 94% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 95% identity with respect to sequence number 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 96% identity with respect to SEQ ID NO. 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 97% identity with respect to SEQ ID NO. 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 98% identity with respect to SEQ ID NO. 13. In an embodiment, the heavy chain variable domain comprises a sequence having at least 99% identity with respect to SEQ ID NO. 13. In an embodiment, the heavy chain variable domain comprises SEQ ID NO. 13. In an embodiment, the heavy chain variable domain is SEQ ID NO. 13.

[0129] In an embodiment, the light chain variable domain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 90% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 91% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 92% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 93% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 94% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 95% identity with respect to sequence number 14. In an embodiment, the light chain variable domain comprises a sequence having at least 96% identity with respect to SEQ ID NO. 14. In an embodiment, the light chain variable domain comprises a sequence having at least 97% identity with respect to SEQ ID NO. 14. In an embodiment, the light chain variable domain comprises a sequence having at least 98% identity with respect to SEQ ID NO. 14. In an embodiment, the light chain variable domain comprises a sequence having at least 99% identity with respect to SEQ ID NO. 14. In an embodiment, the light chain variable domain comprises SEQ ID NO. 14. In an embodiment, the light chain variable domain is SEQ ID NO. 14.

[0130] In an embodiment, the heavy chain variable domain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence no. 13, and the light chain variable domain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence no. 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 90% identity with respect to sequence no. 13, and the light chain variable domain comprises a sequence having at least 90% identity with respect to sequence no. 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 91% identity with respect to sequence no. 13, and the light chain variable domain comprises a sequence having at least 91% identity with respect to sequence no. 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 92% identity with respect to sequence number 13, and the light chain variable domain comprises a sequence having at least 92% identity with respect to sequence number 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 93% identity with respect to sequence number 13, and the light chain variable domain comprises a sequence having at least 93% identity with respect to sequence number 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 94% identity with respect to sequence number 13, and the light chain variable domain comprises a sequence having at least 94% identity with respect to sequence number 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 95% identity with respect to sequence number 13, and the light chain variable domain comprises a sequence having at least 95% identity with respect to sequence number 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 96% identity with sequence number 13, and the light chain variable domain comprises a sequence having at least 96% identity with sequence number 14.In an embodiment, the heavy chain variable domain comprises a sequence having at least 97% identity with SEQ ID NO. 13, and the light chain variable domain comprises a sequence having at least 97% identity with SEQ ID NO. 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 98% identity with SEQ ID NO. 13, and the light chain variable domain comprises a sequence having at least 98% identity with SEQ ID NO. 14. In an embodiment, the heavy chain variable domain comprises a sequence having at least 99% identity with SEQ ID NO. 13, and the light chain variable domain comprises a sequence having at least 99% identity with SEQ ID NO. 14. In an embodiment, the heavy chain variable domain comprises SEQ ID NO. 13, and the light chain variable domain comprises SEQ ID NO. 14. In an embodiment, the heavy chain variable domain is SEQ ID NO. 13, and the light chain variable domain is SEQ ID NO. 14.

[0131] In an embodiment, the heavy chain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 90% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 91% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 92% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 93% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 94% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 95% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 96% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 97% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 98% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises a sequence having at least 99% identity with respect to SEQ ID NO. 29. In an embodiment, the heavy chain comprises SEQ ID NO. 29. In an embodiment, the heavy chain is SEQ ID NO. 29.

[0132] In an embodiment, the light chain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 90% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 91% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 92% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 93% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 94% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 95% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 96% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 97% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 98% identity with respect to sequence no. 30. In an embodiment, the light chain comprises a sequence having at least 99% identity with respect to sequence no. 30. In an embodiment, the light chain comprises sequence no. 30. In an embodiment, the light chain is sequence no. 30.

[0133] In an embodiment, the heavy chain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 90% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 90% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 91% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 91% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 92% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 92% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 93% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 93% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 94% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 94% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 95% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 95% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 96% identity with respect to sequence no. 29, and the light chain comprises a sequence having at least 96% identity with respect to sequence no. 30. In an embodiment, the heavy chain comprises a sequence having at least 97% identity with respect to sequence number 29, and the light chain comprises a sequence having at least 97% identity with respect to sequence number 30.In an embodiment, the heavy chain comprises a sequence having at least 98% identity with respect to SEQ ID NO. 29, and the light chain comprises a sequence having at least 98% identity with respect to SEQ ID NO. 30. In an embodiment, the heavy chain comprises a sequence having at least 99% identity with respect to SEQ ID NO. 29, and the light chain comprises a sequence having at least 99% identity with respect to SEQ ID NO. 30. In an embodiment, the heavy chain comprises SEQ ID NO. 29, and the light chain comprises SEQ ID NO. 30. In an embodiment, the heavy chain is SEQ ID NO. 29, and the light chain is SEQ ID NO. 30.

[0134] In an embodiment, the antibody is IgG. In an embodiment, the antibody is IgG1 or IgG2. In an embodiment, the antibody is IgG1. In an embodiment, the antibody is IgG2. In an embodiment, the antibody is humanized IgG1. In an embodiment, the antibody is humanized IgG2.

[0135] In an embodiment, the antibody is a humanized antibody or a chimeric antibody. In an embodiment, the antibody is a humanized antibody. In an embodiment, the antibody is a chimeric antibody.

[0136] As described above, in an embodiment, the antibody may be a fragment of an antibody. In an embodiment, the antibody comprises a Fab fragment. In an embodiment, the antibody is a single-chain antibody (scFv). In an embodiment, the light chain variable domain and the heavy chain variable domain form part of the scFv. In an embodiment, the antibody comprises a fragment crystallizable (Fc) domain. In an embodiment, the antibody is a single-domain antibody. In an embodiment, the single-domain antibody comprises a light chain variable domain. In an embodiment, the single-domain antibody comprises a heavy chain variable domain.

[0137] The antibody provided herein may include one or more glycosylation sites within the IgG heavy chain. For example, the glycosylation site may be located within the variable heavy domain of the antibody. In an embodiment, the Fc domain of the antibody includes one or more glycosylation sites. In an embodiment, the antibody provided herein includes one or more modifications that remove potential glycosylation sites. In an embodiment, the Fc domain may include one or more modifications in which one or more carbohydrate moietys are added, substituted, deleted, or modified. For example, substitution of residues with asparagine-X-serine or asparagine-X-threonine motifs may create a potential site for enzymatic attachment of the carbohydrate moiety and thus may be used to regulate the glycosylation of the antibody. In an embodiment, one or more glycosylation sites may be associated with or enhance the binding of the antibody to Fc receptors (e.g., FcγRI, FcγRII, FcγRII). Accordingly, in the embodiments, the presence of one or more glycosylation sites in the antibody may enhance, for example, antibody-dependent cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC). In the embodiments, the Fc domain comprises one or more modifications to reduce binding to the Fc receptor. In the embodiments, the Fc domain may comprise one or more modifications in which one or more glycosylation sites are modified or deleted. In the embodiments, one or more modifications include the removal of the glycosylation site.

[0138] In an embodiment, the antibody provided herein comprising an embodiment thereof may comprise a modified Fc domain that does not induce antibody-dependent cytotoxicity (ADCC). In an embodiment, the antibody provided herein comprising an embodiment thereof comprises an Fc portion that does not induce ADCC in the presence of effector cells (e.g., cytotoxic T cells). In an embodiment, the antibody provided herein comprising an embodiment thereof may comprise a modified Fc domain that does not induce complement-dependent cytotoxicity (CDC). In an embodiment, the Fc domain comprises effector cells that inhibit substitution. In the presence of effector cells that inhibit substitution, binding of the Fc domain to an effector cell ligand reduces the activation of the effector cells compared to the absence of said substitution. In an embodiment, binding of the Fc domain to an effector cell ligand does not substantially cause activation of the effector cells compared to the absence of said substitution.

[0139] In an embodiment, an antibody may bind to the WISP1 protein. In an embodiment, the WISP1 protein is a mammalian WISP1 protein. In an embodiment, the WISP1 protein is a human WISP1 protein. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 15, 16, or 17. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 19. In an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 20. In an embodiment, the WISP1 protein is the sequence of SEQ ID NO. 19. In an embodiment, the WISP1 protein is the sequence of SEQ ID NO. 20. In an embodiment, the antibody may bind to domain 2 of WISP1. In an embodiment, the WISP1 protein comprises domain 2. In an embodiment, domain 2 comprises the sequence of SEQ ID NO. 26. Accordingly, in an embodiment, the WISP1 protein comprises the sequence of SEQ ID NO. 26. In an embodiment, the WISP1 protein is the sequence of SEQ ID NO. 26.

[0140] In an embodiment, the antibody may bind to domain 2 of the WISP1 protein. In an embodiment, the domain 2 of the WISP1 protein comprises residues corresponding to positions 121 to 203 of SEQ ID NO. 19. In an embodiment, the antibody may bind to residues within the region corresponding to positions 121 to 203 of SEQ ID NO. 19.

[0141] The anti-WISP1 antibody provided herein, including embodiments thereof, exhibits high affinity binding and specificity. In the embodiments, the anti-WISP1 antibody has the advantage of being cross-reactive to human and non-human primate WISP1 (e.g., cynomolgus WISP1), thereby facilitating production and preclinical testing. Accordingly, in the embodiments, the antibody binds to human WISP1 or cynomolgus WISP1.

[0142] The ability of an antibody to bind to a specific epitope (e.g., WISP1 protein, domain 2 of the WISP1 protein) is equal to the equilibrium dissociation constant (K D It can be described by ). Equilibrium dissociation constant (K as defined herein). D ) is the ratio of the dissociation rate (K-off) to the binding rate (K-on) of the antibody against the WISP1 protein. This is expressed by the following formula: K D = K-off / K-on.

[0143] In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 1 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 1.5 nM to about 20 nM. DIt binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 2 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 2.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 3 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 3.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 4 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 4.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 5 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 5.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 6 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 6.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 7 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 7.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 8 nM to about 20 nM. D) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 8.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 9 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 9.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 10 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 10.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 11 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 11.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 12 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 12.5 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 13 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 13.5 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 14 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 14.5 nM to about 20 nM. DIt binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 15 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 15.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 16 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 16.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 17 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 17.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 18 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 18.5 nM to about 20 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 19 nM to about 20 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 19.5 nM to about 20 nM. D It binds to the WISP1 protein.

[0144] In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 19.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 19 nM. DIt binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 18.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 18 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 17.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 17 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 16.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 16 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 15.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 15 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 14.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 14 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 13.5 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 13 nM. DIt binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 12.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 12 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 11.5 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 11 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 10.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 10 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 9.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 9 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 8.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 8 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 7.5 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 7 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 6.5 nM.D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 6 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 5.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 4.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 4 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 3.5 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 3 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 2.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 2 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 1.5 nM. D ) binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 1 nM. D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 0.5 nM. D It binds to the WISP1 protein.

[0145] In the examples, the antibody is about 0.01, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, or 20 nM of K D It binds to the WISP1 protein.

[0146] In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 0.1 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.9 nM to 1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1 nM to about 10 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 1.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 1.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 1.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 2.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 2.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.5 nM to about 10 nM of KD It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 2.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 4.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 4.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 4.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5 nM to about 10 nM of KD It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 5.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 5.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 5.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 6.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 6.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.4 nM to about 10 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 6.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 6.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 7.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 7.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 7.9 nM to about 10 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 8.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 8.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.4 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 8.9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody has an equilibrium dissociation constant (K) of about 9.1 nM to about 10 nM. D It binds to the WISP1 protein via ). In an embodiment, the antibody contains about 9.2 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.3 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.4 nM to about 10 nM of KD It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.5 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.6 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.7 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.8 nM to about 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 9.9 nM to about 10 nM of K D It binds to the WISP1 protein.

[0147] In an example, the antibody is about 0.01 nM to about 9.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9.1 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.7 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.3 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.9 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.5 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2.1 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.6 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 1.1 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.9 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.8 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.7 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.6 nM of K DIt binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.5 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.4 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.3 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.2 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody contains about 0.01 nM to about 0.1 nM of K D It binds to the WISP1 protein.

[0148] In the examples, the antibody is about 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10 nM of K D It binds to the WISP1 protein.

[0149] In an example, the antibody is about 5.55 nM K DIt binds to the WISP1 protein. In an embodiment, the antibody is antibody K4, and K at a concentration of 0.01 nM to about 20 nM. D The antibody K4 binds to the WISP1 protein. In an example, the antibody K4 is about 0.01, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, or 20 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody is antibody K4, and K at a concentration of 0.01 nM to about 10 nM. D It binds to the WISP1 protein. In an example, antibody K4 is 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 nM of K D It binds to the WISP1 protein. In an embodiment, the antibody is antibody K4, and 5.55 nM of K D It binds to the WISP1 protein.

[0150] In one embodiment, the antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ NO. 1, CDR H2 as presented in SEQ NO. 2; and CDR H3 as presented in SEQ NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ NO. 4, CDR L2 as presented in AAS, and CDR L3 as presented in SEQ NO. 6. In one embodiment, the antibody comprises a heavy chain variable domain of SEQ NO. 13 and a light chain variable domain of SEQ NO. 14. In one additional embodiment, the antibody is antibody K4.

[0151] In an embodiment, the antibody is attached to a therapeutic agent. In an embodiment, the antibody is attached to a diagnostic agent. In an embodiment, the diagnostic agent is a detectable moiety.

[0152] In one embodiment, an anti-WISP1 antibody is provided. The anti-WISP1 antibody binds to the same epitope as the anti-WISP1 antibody, comprising a heavy chain variable domain including CDR H1 as presented in SEQ NO. 1, CDR H2 as presented in SEQ NO. 2, and CDR H3 as presented in SEQ NO. 3; and a light chain variable domain including CDR L1 as presented in SEQ NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ NO. 6. In an embodiment, the antibody comprises a heavy chain variable domain including SEQ NO. 13 and a light chain variable domain including SEQ NO. 14.

[0153] In one embodiment, an anti-WISP1 antibody is provided. The antibody competes with the anti-WISP1 antibody for binding to the WISP1 protein, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 as presented in SEQ ID NO. 4, and CDR L3 as presented in SEQ ID NO. 6. In an embodiment, the antibody comprises a heavy chain variable domain comprising SEQ ID NO. 13 and a light chain variable domain comprising SEQ ID NO. 14.

[0154] In an embodiment, the antibody can bind to the WISP1 protein. In an embodiment, the WISP1 protein is a mammalian WISP1 protein. In an embodiment, the WISP1 protein is a human WISP1 protein. In an embodiment, the antibody binds to the WISP1 protein. In an embodiment, the WISP1 protein comprises the amino acid sequence of SEQ ID NO. 19. In an embodiment, the WISP1 protein comprises the amino acid sequence of SEQ ID NO. 20. In an embodiment, the WISP1 protein is the amino acid sequence of SEQ ID NO. 19. In an embodiment, the WISP1 protein is the amino acid sequence of SEQ ID NO. 20. In an embodiment, the antibody can bind to domain 2 of the WISP1 protein.

[0155] In an embodiment, the anti-WISP1 antibody provided herein comprising an embodiment thereof inhibits WISP1 signaling or activity compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, WISP1 signaling or activity may be evaluated by the expression of fibrosis-promoting or inflammatory genes (e.g., IL-6, pSMAD2, pSMAD3, Col1a1, Col3a1, Fn1, Tgfb1, and Ccn2, etc.). In an embodiment, WISP1 signaling or activity may be evaluated by collagen content or hydroxyproline production. In an embodiment, WISP1 signaling or activity may be evaluated by fibroblast chemotaxis or migration. In an embodiment, the anti-WISP1 antibody inhibits WISP1 signaling or activity by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody may reduce WISP1 signaling or activity by 10% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody may reduce WISP1 signaling or activity by 20% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody may reduce WISP1 signaling or activity by 30% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 40% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 50% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody.In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 60% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 70% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 80% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 90% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by 95% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody can reduce WISP1 signaling or activity by more than 95% compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody.

[0156] In an embodiment, the anti-WISP1 antibody provided herein comprising an embodiment thereof may reduce WISP1 signaling or activity by 1.5 times, 2 times, 3 times, 4 times, 5 times, 10 times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein comprising an embodiment thereof may reduce WISP1 signaling or activity by 1.5 times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein comprising an embodiment thereof may reduce WISP1 signaling or activity by 2 times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein, comprising an embodiment thereof, can reduce WISP1 signaling or activity by three times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein, comprising an embodiment thereof, can reduce WISP1 signaling or activity by four times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein, comprising an embodiment thereof, can reduce WISP1 signaling or activity by five times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody. In an embodiment, the anti-WISP1 antibody provided herein, comprising an embodiment thereof, can reduce WISP1 signaling or activity by ten times or less compared to WISP1 signaling or activity in the absence of the anti-WISP1 antibody.

[0157] nucleic acid composition

[0158] The compositions provided herein comprise a nucleic acid molecule encoding the anti-WISP1 antibody or a fragment thereof provided herein, comprising embodiments thereof. Antibodies encoded by isolated nucleic acids are described in detail throughout this application (including the detailed description and examples section above). Accordingly, in one embodiment, isolated nucleic acids encoding an antibody such as provided herein, comprising embodiments thereof, are provided.

[0159] In an embodiment, the isolated nucleic acid codes for a variable heavy chain domain or a variable light chain domain provided herein. In an embodiment, the isolated nucleic acid codes for a variable heavy chain domain. In an embodiment, the isolated nucleic acid codes for a variable light chain domain.

[0160] In an embodiment, the isolated nucleic acid may be provided to a vector, e.g., an expression vector. Accordingly, a vector comprising the isolated nucleic acid provided herein, comprising embodiments thereof in other aspects, is provided. In an embodiment, the vector is an expression vector capable of inducing the expression of the nucleic acid to which it is operablely linked. The term “operably linked” means that the nucleotide sequence of interest is linked to a regulatory sequence(s) that allow the expression of the nucleotide sequence. The regulatory sequence may include, for example, a promoter, an enhancer, and other expression regulatory elements (e.g., a polyadenylation signal). Such regulatory sequences are well known in the art and are described, for example, in the literature [Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990)], the full text of which is incorporated herein for all purposes.

[0161] method

[0162] The composition provided herein (e.g., anti-WISP1 antibody) comprising an embodiment thereof is considered to provide an effective treatment for a disease such as fibrosis (e.g., hepatic fibrosis, pulmonary fibrosis, cardiac fibrosis, etc.). Accordingly, in one embodiment, a method for treating fibrosis in a subject requiring treatment for fibrosis is provided, said method comprising the step of administering a therapeutically effective amount of antibody to said subject.

[0163] In an embodiment, fibrosis is renal fibrosis, fibrotic liver disease, pulmonary fibrosis, cardiac fibrosis, bone or bone marrow fibrosis, or cutaneous fibrosis. In an embodiment, fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis. In an embodiment, fibrosis is fibrotic liver disease. In an embodiment, fibrotic liver disease may be referred to as "liver fibrosis." In an embodiment, fibrosis is pulmonary fibrosis. In an embodiment, lung fibrosis may be referred to as "pulmonary fibrosis." In an embodiment, fibrosis is cardiac fibrosis. In an embodiment, cardiac fibrosis may be referred to as "heart fibrosis." In an embodiment, pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF). In an embodiment, fibrosis is cutaneous fibrosis. In an embodiment, fibrosis is renal fibrosis. In an embodiment, fibrosis is bone fibrosis. In an embodiment, fibrosis is bone marrow fibrosis.

[0164] In an embodiment, the subject has interstitial lung disease. In an embodiment, the interstitial lung disease causes pulmonary fibrosis in the subject. For example, if the interstitial lung disease causes scarring of lung tissue, the subject has pulmonary fibrosis. In an embodiment, the interstitial lung disease and / or pulmonary fibrosis may be idiopathic, autoimmune-related, connective tissue disease-related, environmental, direct, radiation-induced, or drug-induced. In an embodiment, the interstitial lung disease and / or pulmonary fibrosis may be associated with a viral infection or lymphangioleiomyomatosis. In an embodiment, the interstitial lung disease and / or pulmonary fibrosis may be hereditary.

[0165] In an embodiment, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis. In an embodiment, the interstitial lung disease is non-specific interstitial pneumonia. In an embodiment, the interstitial lung disease is rheumatoid arthritis interstitial lung disease. In an embodiment, the interstitial lung disease is hypersensitivity pneumonia. In an embodiment, the interstitial lung disease is beryllium. In an embodiment, the interstitial lung disease is respiratory bronchiolitis interstitial lung disease. In an embodiment, the interstitial lung disease is desquamative interstitial pneumonia. In an embodiment, the interstitial lung disease is sarcoidosis. In an embodiment, the interstitial lung disease is acute interstitial pneumonia. In an embodiment, the interstitial lung disease is idiopathic organizing pneumonia. In an embodiment, the interstitial lung disease is lymphocytic interstitial pneumonia. In an embodiment, the interstitial lung disease is pleural parenchymal elastic fibrosis.

[0166] In an embodiment, pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis. In an embodiment, pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0167] In an embodiment, pulmonary fibrosis is idiopathic pulmonary fibrosis. In an embodiment, pulmonary fibrosis is systemic sclerosis-related pulmonary fibrosis. In an embodiment, pulmonary fibrosis is progressive pulmonary fibrosis. In an embodiment, pulmonary fibrosis is Sjögren's syndrome-related pulmonary fibrosis. In an embodiment, pulmonary fibrosis is pneumoconiosis. In an embodiment, pulmonary fibrosis is asbestosis. In an embodiment, pulmonary fibrosis is silicosis. In an embodiment, pulmonary fibrosis is radiation-induced pulmonary fibrosis. In an embodiment, pulmonary fibrosis is drug-induced pulmonary fibrosis. In an embodiment, pulmonary fibrosis is COVID-19-related pulmonary fibrosis. In an embodiment, pulmonary fibrosis is pulmonary fibrosis after acute respiratory distress syndrome. In an embodiment, pulmonary fibrosis is lymphangioleiomyomatosis-related pulmonary fibrosis. In an embodiment, pulmonary fibrosis is familial pulmonary fibrosis. In an embodiment, pulmonary fibrosis is Hermanski-Pudlak syndrome-related pulmonary fibrosis. In an embodiment, pulmonary fibrosis is congenital dyskeratosis-related pulmonary fibrosis.

[0168] In an embodiment, fibrosis is associated with autoimmunity. For example, in an embodiment, the subject has an autoimmune disorder that causes the overproduction of collagen, leading to fibrosis. In an embodiment, the subject has scleroderma.

[0169] In an embodiment, the method further comprises the step of identifying, in a biological sample obtained from a subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof.

[0170] A method for treating fibrosis in a subject requiring treatment for fibrosis in another embodiment is provided, said method comprising the following steps: a) identifying, in a biological sample obtained from said subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) a step of administering a therapeutically effective amount of the anti-WISP1 antibody provided herein, including an embodiment thereof, to the subject.

[0171] In an embodiment, the method includes the step of identifying an increased level of WISP1 compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method includes the step of identifying an increased level of α-smooth muscle actin (α-SMA) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method includes the step of identifying an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method includes the step of identifying an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method includes the step of identifying an increased level of fibronectin (Fn1) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method includes the step of identifying an increased level of interleukin 6 (IL-6) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method comprises the step of identifying an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the method comprises the step of identifying an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control in a biological sample obtained from a subject. In an embodiment, the standard control is a biological sample obtained from a subject without fibrosis. In an embodiment, the standard control comprises cells derived from liver tissue of a subject without fibrosis. In an embodiment, the standard control comprises cells derived from lung tissue of a subject without fibrosis.

[0172] In an embodiment, fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis. In an embodiment, fibrosis is fibrotic liver disease. In an embodiment, fibrosis is pulmonary fibrosis. In an embodiment, fibrosis is cardiac fibrosis.

[0173] In an embodiment, the subject has interstitial lung disease. In an embodiment, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0174] In an embodiment, pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0175] In the method provided herein, in the embodiments, pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis. In the embodiments, pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis. In the embodiments, pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0176] In another embodiment, a method is provided for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, said method comprising the step of contacting a cell with an anti-WISP1 antibody provided herein, comprising embodiments thereof. In an embodiment, said method comprises inhibiting the release of IL-6 from the cell. In an embodiment, said method comprises inhibiting the release of pSMAD2 from the cell. In an embodiment, said method comprises inhibiting the release of pSMAD3 from the cell.

[0177] In an embodiment, the cell is an immune cell. In an embodiment, the cell is a fibroblast.

[0178] A method for inhibiting the migration of WISP-1 receptor-expressing cells is provided in another embodiment, said method comprising the step of contacting said cells with an anti-WISP1 antibody provided herein, comprising an embodiment thereof. As used herein, "WISP-1 receptor" refers to a protein that binds to a WISP-1 protein. In an embodiment, the WISP-1 receptor is an integrin.

[0179] In an embodiment, the cell is a fibroblast or an astrocyte. In an embodiment, the cell is a fibroblast. In an embodiment, the cell is an astrocyte. In an embodiment, the astrocyte is a hepatic astrocyte.

[0180] In one embodiment, a method for detecting WISP1-expressing cells is provided, said method comprising: (i) contacting WISP1-expressing cells with an antibody provided herein comprising an embodiment thereof; and (ii) detecting the binding of the antibody to a WISP1 protein expressed by said cells. In an embodiment, the antibody is attached to a detectable moiety.

[0181] In one embodiment, there exists a method for delivering a therapeutic agent to WISP1-expressing cells, said method comprising the step of contacting the WISP1-expressing cells with an antibody provided herein, which includes an embodiment thereof, wherein the antibody is attached to the therapeutic agent. In an embodiment, the therapeutic agent is an anti-fibrotic agent.

[0182] In the method provided herein, in an embodiment, contact occurs in vitro. In an embodiment, WISP1-expressing cells are in a subject. In an embodiment, the subject is a healthy subject. In an embodiment, the subject is a subject having fibrosis. In an embodiment, the fibrosis is pulmonary fibrosis or hepatic fibrosis. In an embodiment, the fibrosis is pulmonary fibrosis. In an embodiment, the fibrosis is hepatic fibrosis.

[0183] The embodiments and modes of embodiment described herein are for illustrative purposes only, and various modifications or variations related thereto are to be proposed to those skilled in the art and are understood to be included within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

[0184] P Implementation mode

[0185] Embodiment P1. An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and the light chain variable domain comprises CDR L1 as presented in SEQ ID NO. 4, CDR L2 as presented in SEQ ID NO. 5, and CDR L3 as presented in SEQ ID NO. 6.

[0186] Embodiment P2. The antibody of Embodiment P1, wherein the heavy chain variable domain comprises a sequence having at least 90% identity with respect to Sequence No. 13.

[0187] Embodiment P3. An antibody in Embodiment P1 or P2, wherein the heavy chain variable domain comprises a sequence having at least 95% identity with respect to Sequence No. 13.

[0188] Embodiment P4. An antibody in any one of embodiments P1 to P3, wherein the heavy chain variable domain comprises a sequence having at least 98% identity with respect to sequence number 13.

[0189] Embodiment P5. In any one of embodiments P1 to P4, the heavy chain variable domain is an antibody comprising sequence number 13.

[0190] Embodiment P6. An antibody in any one of embodiments P1 to P5, wherein the light chain variable domain comprises a sequence having at least 90% identity with respect to sequence number 14.

[0191] Embodiment P7. An antibody in any one of embodiments P1 to P6, wherein the light chain variable domain comprises a sequence having at least 95% identity with respect to sequence number 14.

[0192] Embodiment P8. An antibody in any one of embodiments P1 to P7, wherein the light chain variable domain comprises a sequence having at least 98% identity with respect to sequence number 14.

[0193] Embodiment P9. In any one of embodiments P1 to P8, the light chain variable domain is an antibody comprising SEQ ID NO. 14.

[0194] Embodiment P10. In any one of Embodiments P1 to P9, the antibody is an IgG antibody.

[0195] Embodiment P11. In any one of embodiments P1 to P10, the antibody is a humanized antibody or a chimeric antibody.

[0196] Embodiment P12. In any one of Embodiments P1 to P11, the antibody is an antibody comprising a Fab fragment.

[0197] Embodiment P13. In any one of Embodiments P1 to P11, the antibody is a single-chain antibody (scFv).

[0198] Embodiment P14. In any one of Embodiments P1 to P13, the antibody is an antibody capable of binding to the WISP1 protein.

[0199] Embodiment P15. In Embodiment P14, the WISP1 protein is an antibody that is a human WISP1 protein.

[0200] Embodiment P16. In Embodiment P14 or P15, the WISP1 protein is an antibody comprising the sequence of SEQ ID NO. 19 or 20.

[0201] Embodiment P17. In any one of embodiments P14 to P16, the antibody is an antibody capable of binding to domain 2 of the WISP1 protein.

[0202] Embodiment P18. In any one of embodiments P14 to P17, the domain 2 of the WISP1 protein comprises residues corresponding to 121 to 203 of sequence number 19.

[0203] Embodiment P19. In any one of embodiments P14 to P18, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 20 nM. D An antibody capable of binding to the above WISP1 protein.

[0204] Embodiment P20. In Embodiment P19, the antibody is an antibody capable of binding to the WISP1 protein with a KD of about 5.55 nM.

[0205] Embodiment P21. An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 as presented in SEQ ID NO. 5, and CDR L3 as presented in SEQ ID NO. 6, and an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody that binds to the same epitope as the anti-WISP1 antibody.

[0206] Embodiment P22. An antibody competing with an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody for binding to a WISP1 protein, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 as presented in SEQ ID NO. 5, and CDR L3 as presented in SEQ ID NO. 6.

[0207] Embodiment P23. In any one of Embodiments P1 to P22, the antibody is an antibody attached to a therapeutic agent.

[0208] Embodiment P24. In any one of embodiments P1 to P22, the antibody is an antibody attached to a diagnostic agent.

[0209] Embodiment P25. A method for treating fibrosis in a subject requiring treatment for fibrosis, comprising the step of administering a therapeutically effective amount of any one of embodiments P1 to P24 to the subject.

[0210] Embodiment P26. The method of Embodiment P25, wherein the fibrosis is a fibrotic liver disease.

[0211] Embodiment P27. The method of Embodiment P25, wherein the fibrosis is pulmonary fibrosis.

[0212] Embodiment P28. A method for treating fibrosis in a subject requiring treatment for fibrosis, the method comprising: a) identifying, in a biological sample obtained from the subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) a step of administering a therapeutically effective amount of any one of embodiments P1 to P24 of an anti-WISP1 antibody to the subject.

[0213] Embodiment P29. The method of Embodiment P28, wherein the fibrosis is a fibrotic liver disease.

[0214] Embodiment P30. The method of Embodiment P28, wherein the fibrosis is pulmonary fibrosis.

[0215] Embodiment P31. A method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, wherein the method comprises the step of contacting the cell with an anti-WISP1 antibody of any one of Embodiments P1 to P24.

[0216] Embodiment P32. The method of Embodiment P31, wherein the cell is an immune cell.

[0217] Embodiment P33. The method of Embodiment P31, wherein the cell is a fibroblast.

[0218] Embodiment P34. A method for inhibiting the migration of WISP-1 receptor-expressing cells, comprising the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments P1 to P24.

[0219] Embodiment P35. The method of Embodiment P34, wherein the WISP-1 receptor is an integrin.

[0220] Embodiment P36. The method in Embodiment P34 or P35, wherein the cell is a fibroblast or an astrocyte.

[0221] Embodiment P37. A method for detecting WISP1-expressing cells, the method comprising the following steps: (i) contacting WISP1-expressing cells with an antibody of any one of Embodiments P1 to P24; and (ii) detecting the binding of said antibody to a WISP1 protein expressed by said cells.

[0222] Embodiment P38. The method of Embodiment P37, wherein the antibody is attached to a detectable moiety.

[0223] Mode of implementation

[0224] Embodiment 1. An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and the light chain variable domain comprises CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0225] Embodiment 2. The antibody of Embodiment 1, wherein the heavy chain variable domain comprises a sequence having at least 90% identity with respect to Sequence No. 13.

[0226] Embodiment 3. An antibody according to Embodiment 1 or 2, wherein the heavy chain variable domain comprises a sequence having at least 95% identity with respect to Sequence No. 13.

[0227] Embodiment 4. An antibody in any one of Embodiments 1 to 3, wherein the heavy chain variable domain comprises a sequence having at least 98% identity with respect to Sequence No. 13.

[0228] Embodiment 5. In any one of Embodiments 1 to 4, the heavy chain variable domain is an antibody comprising SEQ ID NO. 13.

[0229] Embodiment 6. An antibody in any one of Embodiments 1 to 5, wherein the light chain variable domain comprises a sequence having at least 90% identity with respect to Sequence No. 14.

[0230] Embodiment 7. An antibody in any one of Embodiments 1 to 6, wherein the light chain variable domain comprises a sequence having at least 95% identity with respect to Sequence No. 14.

[0231] Embodiment 8. An antibody in any one of Embodiments 1 to 7, wherein the light chain variable domain comprises a sequence having at least 98% identity with respect to Sequence No. 14.

[0232] Embodiment 9. In any one of Embodiments 1 to 8, the light chain variable domain is an antibody comprising SEQ ID NO. 14.

[0233] Embodiment 10. In any one of Embodiments 1 to 9, the antibody is an IgG antibody.

[0234] Embodiment 11. In any one of Embodiments 1 to 10, the antibody is a humanized antibody or a chimeric antibody.

[0235] Embodiment 12. In any one of Embodiments 1 to 11, the antibody is an antibody comprising a Fab fragment.

[0236] Embodiment 13. P2 In any one of Embodiments 1 to 11, the antibody is a single-chain antibody (scFv).

[0237] Embodiment 14. In any one of Embodiments 1 to 13, the antibody is an antibody capable of binding to the WISP1 protein.

[0238] Embodiment 15. In Embodiment 14, the WISP1 protein is an antibody that is a human WISP1 protein.

[0239] Embodiment 16. In Embodiment 14 or 15, the WISP1 protein is an antibody comprising the sequence of SEQ ID NO. 19 or 20.

[0240] Embodiment 17. In any one of Embodiments 14 to 16, the antibody is an antibody capable of binding to domain 2 of the WISP1 protein.

[0241] Embodiment 18. In any one of Embodiments 14 to 17, the domain 2 of the WISP1 protein comprises residues corresponding to 121 to 203 of SEQ ID NO. 19.

[0242] Embodiment 19. In any one of Embodiments 14 to 18, the antibody has an equilibrium dissociation constant (K) of about 0.01 nM to about 20 nM. D An antibody capable of binding to the above WISP1 protein.

[0243] Embodiment 20. In Embodiment 19, the antibody is an antibody capable of binding to the WISP1 protein with a KD of about 5.55 nM.

[0244] Embodiment 21. An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 comprising AAS, and CDR L3 as presented in SEQ ID NO. 6, and an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody that binds to the same epitope as the anti-WISP1 antibody.

[0245] Embodiment 22. An antibody competing with an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody for binding to a WISP1 protein, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 comprising AAS, and CDR L3 as presented in SEQ ID NO. 6.

[0246] Embodiment 23. In any one of Embodiments 1 to 22, the antibody is an antibody attached to a therapeutic agent.

[0247] Embodiment 24. In any one of Embodiments 1 to 22, the antibody is an antibody attached to a diagnostic agent.

[0248] Embodiment 25. A method for treating fibrosis in a subject requiring treatment for fibrosis, comprising the step of administering a therapeutically effective amount of any one of Embodiments 1 to 24 to the subject.

[0249] Embodiment 26. The method of Embodiment 25, wherein the fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0250] Embodiment 27. The method of Embodiment 26, wherein the fibrosis is a fibrotic liver disease.

[0251] Embodiment 28. The method of Embodiment 26, wherein the fibrosis is pulmonary fibrosis.

[0252] Embodiment 29. The method of Embodiment 28, wherein the subject has interstitial lung disease.

[0253] Embodiment 30. The method of Embodiment 29, wherein the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0254] Embodiment 31. A method in any one of embodiments 28 to 30, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0255] Embodiment 32. The method of Embodiment 31, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0256] Embodiment 33. The method of Embodiment 32, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0257] Embodiment 34. The method of Embodiment 26, wherein the fibrosis is cardiac fibrosis.

[0258] Embodiment 35. A method in which, in any one of embodiments 25-34, the subject has scleroderma.

[0259] Embodiment 36. A method for treating fibrosis in a subject requiring treatment for fibrosis, the method comprising: a) identifying, in a biological sample obtained from the subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) a step of administering a therapeutically effective amount of any one of embodiments 1 to 24 of an anti-WISP1 antibody to the subject.

[0260] Embodiment 37. The method of Embodiment 36, wherein the fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0261] Embodiment 38. The method of Embodiment 36, wherein the fibrosis is a fibrotic liver disease.

[0262] Embodiment 39. The method of Embodiment 36, wherein the fibrosis is pulmonary fibrosis.

[0263] Embodiment 40. The method of Embodiment 39, wherein the subject has interstitial lung disease.

[0264] Embodiment 41. The method according to Embodiment 40, wherein the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0265] Embodiment 42. A method in any one of embodiments 39 to 41, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0266] Embodiment 43. The method of Embodiment 42, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0267] Embodiment of Implementation 44. In embodiment 43, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0268] Embodiment 45. The method of Embodiment 37, wherein the fibrosis is cardiac fibrosis.

[0269] Embodiment 46. A method in any one of embodiments 36 to 45, wherein the subject has scleroderma.

[0270] Embodiment 47. A method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, the method comprising the step of contacting the cell with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0271] Embodiment 48. The method of Embodiment 47, wherein the cell is an immune cell.

[0272] Embodiment 49. The method of Embodiment 47, wherein the cell is a fibroblast.

[0273] Embodiment 50. A method for inhibiting the migration of WISP-1 receptor-expressing cells, wherein the method comprises the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0274] Embodiment 51. A method in which, in Embodiment 50, the WISP-1 receptor is an integrin.

[0275] Embodiment 52. The method of Embodiment 50 or 51, wherein the cell is a fibroblast or an astrocyte.

[0276] Embodiment 53. A method for detecting WISP1-expressing cells, the method comprising: i) contacting WISP1-expressing cells with an antibody of any one of Embodiments 1 to 24; and ii) detecting the binding of said antibody to a WISP1 protein expressed by said cells.

[0277] Embodiment 54. The method of Embodiment 53 in which the antibody is attached to a detectable moiety.

[0278] Embodiment 55. An antibody according to any one of Embodiments 1 to 24 for use in a method of treating fibrosis in a subject requiring treatment for fibrosis.

[0279] Embodiment 56. In Embodiment 55, the fibrosis is an antibody that is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0280] Embodiment 57. In Embodiment 56, the fibrosis is an antibody that is a fibrotic liver disease.

[0281] Embodiment 58. In Embodiment 56, the fibrosis is an antibody that is pulmonary fibrosis.

[0282] Embodiment 59. In Embodiment 58, the subject is an antibody having interstitial lung disease.

[0283] Embodiment 60. In Embodiment 59, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0284] Embodiment 61. In any one of embodiments 58 to 60, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0285] Embodiment 62. In Embodiment 61, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0286] Embodiment 63. In Embodiment 62, the pulmonary fibrosis is an antibody that is idiopathic pulmonary fibrosis.

[0287] Embodiment 64. In Embodiment 56, the fibrosis is an antibody that is cardiac fibrosis.

[0288] Embodiment 65. In any one of Embodiments 55-64, the subject is an antibody having scleroderma.

[0289] Embodiment 66. An antibody according to any one of Embodiments 1 to 24 for use in a method for treating fibrosis in a subject requiring treatment for fibrosis, wherein the method comprises: a) confirming, in a biological sample obtained from the subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) an antibody comprising the step of administering a therapeutically effective amount of any one of embodiments 1 to 24 of an anti-WISP1 antibody to the subject.

[0290] Embodiment 67. In Embodiment 66, the fibrosis is an antibody that is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0291] Embodiment 68. In Embodiment 66, the fibrosis is an antibody that is a fibrotic liver disease.

[0292] Embodiment 69. In Embodiment 66, the fibrosis is an antibody that is pulmonary fibrosis.

[0293] Embodiment 70. In Embodiment 69, the subject is an antibody having interstitial lung disease.

[0294] Embodiment 71. In Embodiment 70, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0295] Embodiment 72. In any one of embodiments 69 to 71, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0296] Embodiment 73. In Embodiment 72, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0297] Embodiment of Implementation 74. In embodiment 73, the pulmonary fibrosis is an antibody that is idiopathic pulmonary fibrosis.

[0298] Embodiment 75. In Embodiment 67, the fibrosis is an antibody that is cardiac fibrosis.

[0299] Embodiment 76. In any one of Embodiments 66 to 75, the subject is an antibody having scleroderma.

[0300] Embodiment 77. An antibody according to any one of Embodiments 1 to 24 for use in a method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, wherein the method comprises the step of contacting the cell with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0301] Embodiment 78. In Embodiment 77, the cell is an antibody that is an immune cell.

[0302] Embodiment 79. In Embodiment 78, the cell is an antibody that is a fibroblast.

[0303] Embodiment 80. An antibody according to any one of Embodiments 1 to 24 for use in a method for inhibiting the migration of WISP-1 receptor-expressing cells, wherein the method comprises the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0304] Embodiment 81. In Embodiment 80, the WISP-1 receptor is an antibody that is an integrin.

[0305] Embodiment 82. In Embodiment 80 or 81, the cell is an antibody that is a fibroblast or an astrocyte.

[0306] Embodiment 83. An antibody according to any one of Embodiments 1 to 24 for use in a method for detecting WISP1-expressing cells, wherein the method comprises: i) contacting a WISP1-expressing cell with an antibody of any one of Embodiments 1 to 24; and ii) detecting the binding of said antibody to a WISP1 protein expressed by said cell.

[0307] Embodiment 84. In Embodiment 83, the antibody is an antibody attached to a detectable moiety.

[0308] Embodiment 85. An in vitro method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from cells, wherein the method comprises the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0309] Embodiment 86. In the in vitro method of Embodiment 85, wherein the cell is an immune cell.

[0310] Embodiment 87. In the in vitro method of Embodiment 85, wherein the cell is a fibroblast.

[0311] Embodiment 88. An in vitro method for inhibiting the migration of WISP-1 receptor-expressing cells, wherein the method comprises the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0312] Embodiment 89. An in vitro method according to Embodiment 88, wherein the WISP-1 receptor is an integrin.

[0313] Embodiment 90. An in vitro method in which the cell is a fibroblast or an astrocyte in Embodiment 88 or 89.

[0314] Embodiment 91. An in vitro method for detecting WISP1-expressing cells, the method comprising: i) contacting WISP1-expressing cells with an antibody of any one of Embodiments 1 to 24; and ii) detecting the binding of said antibody to a WISP1 protein expressed by said cells.

[0315] Embodiment 92. An in vitro method in which the antibody is attached to a detectable moiety in Embodiment 91.

[0316] Embodiment 93. Use of an antibody according to any one of Embodiments 1 to 24 in the manufacture of a medicine for the treatment of fibrosis in a subject requiring treatment thereof.

[0317] Embodiment 94. In Embodiment 93, the fibrosis is used as fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0318] Embodiment 95. In Embodiment 94, the fibrosis is used as a fibrotic liver disease.

[0319] Embodiment 96. In Embodiment 94, the fibrosis is used as pulmonary fibrosis.

[0320] Embodiment 97. In Embodiment 96, the subject is used for interstitial lung disease.

[0321] Embodiment 98. In Embodiment 97, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0322] Embodiment 99. In any one of Embodiments 96 to 98, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0323] Embodiment 100. In Embodiment 99, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0324] Embodiment 101. In Embodiment 100, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0325] Embodiment 102. In Embodiment 94, the fibrosis is cardiac fibrosis.

[0326] Embodiment 103. In any one of Embodiments 93-102, the subject is used for having scleroderma.

[0327] Embodiment 104. Use of an antibody according to any one of Embodiments 1 to 24 in the manufacture of a medicine for treating fibrosis in a subject requiring treatment for fibrosis, wherein the use comprises: a) an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof Use comprising: b) confirming; and b) administering a therapeutically effective amount of any one of embodiments 1 to 24 of an anti-WISP1 antibody to the subject.

[0328] Embodiment 105. In Embodiment 104, the fibrosis is used as fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis.

[0329] Embodiment 106. In Embodiment 104, the fibrosis is used as a fibrotic liver disease.

[0330] Embodiment 107. In Embodiment 104, the fibrosis is used as pulmonary fibrosis.

[0331] Embodiment 108. In Embodiment 107, the subject is used for interstitial lung disease.

[0332] Embodiment 109. In Embodiment 108, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis.

[0333] Embodiment 110. Any one of Embodiments 107 to 109, wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis.

[0334] Embodiment 111. In Embodiment 110, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, or progressive pulmonary fibrosis.

[0335] Embodiment 112. In embodiment 111, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0336] Embodiment 113. In Embodiment 105, the fibrosis is cardiac fibrosis.

[0337] Embodiment 114. In any one of Embodiments 104 to 113, the object has a skin scleroderma.

[0338] Embodiment 115. Use of an antibody according to any one of Embodiments 1 to 24 in the manufacture of a drug for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, wherein the use comprises the step of contacting a cell with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0339] Embodiment 116. In Embodiment 115, the cell is an immune cell.

[0340] Embodiment 117. In Embodiment 115, the cell is a fibroblast.

[0341] Embodiment 118. Use of an antibody according to any one of Embodiments 1 to 24 in the manufacture of a drug for inhibiting the migration of WISP-1 receptor-expressing cells, wherein the use comprises the step of contacting the cells with an anti-WISP1 antibody of any one of Embodiments 1 to 24.

[0342] Embodiment 119. In Embodiment 118, the WISP-1 receptor is used as an integrin.

[0343] Embodiment 120. In Embodiment 118 or 119, the cell is a fibroblast or an astrocyte.

[0344] Embodiment 121. An use of an antibody according to any one of embodiments 1 to 24 in the manufacture of a drug for detecting WISP1-expressing cells, wherein the use comprises: i) contacting WISP1-expressing cells with an antibody of any one of embodiments 1 to 24; and ii) detecting the binding of said antibody to a WISP1 protein expressed by said cells.

[0345] Embodiment 122. In Embodiment 121, the antibody is used to attach to a detectable moiety.

[0346] Embodiment 123. A method in any one of embodiments 25 to 54 in which the object is a human being.

[0347] Embodiment 124. In any one of Embodiments 55 to 76, the antibody is an antibody intended for use in humans.

[0348] Embodiment 125. In any one of Embodiments 93 to 114, the object is a human being.

[0349] Examples

[0350] Example 1: Generation of anti-WISP1 antibody clones

[0351] Anti-human WISP1 antibodies were generated using the Alloy Therapeutics transgenic mouse platform and engineered to express antibodies with a fully human variable domain. Alloy mice, strains ATX-GK and ATX-GK+ were immunized with recombinant human WISP1 (SEQ No. 15) via a rapid immunization method to achieve potent titers within 3 to 5 weeks. Mouse anti-WISP1 titers were determined by ELISA against recombinant human WISP1 (SEQ No. 15), mouse WISP1 (SEQ No. 17), and human CCN2 / CTGF (R&D cat#1680 - SEQ No. 18) as an off-target control. Mice exhibiting potent anti-WISP1 antibody titers were sacrificed, and harvested spleens and lymph nodes were processed by a standard hybridoma fusion procedure.

[0352] After the screening and recovery steps, anti-WISP1 antibody titers were determined in the supernatant of the hybridoma pool, followed by single-cell plating of individual hybridoma clones by FACS sorting. Monoclonal hybridomas were proliferated, and the supernatant was screened by ELISA for binding to recombinant human, synomolgus, and mouse WISP1 (sequence numbers 15, 16, and 17), and counter-screened for binding to recombinant human CCN2 / CTGF (Fig. 1). Hybridoma expression antibodies showing specific binding to the WISP1 protein rather than to CCN2 / CTGF were selected for sequencing.

[0353] Selected monoclonal anti-WISP1 hybridomas were sequenced by the RT-PCR method using the primer set provided for Alloy mice. Duplicate clones were removed by aligning the heavy and light chain variable regions, and clones with associated variable regions were grouped into sequence bins. Unique antibody variable region sequences, e.g., M6-NYS, K4, A1, and A2 (expressed by hybridomas 1D03, 2F09, 3C09, and 9H06, respectively), were subsequently synthesized by the recombinant expression method to generate full-length anti-WISP1 antibodies for further characterization and testing.

[0354] Example 2: Analysis of anti-WISP1 antibody binding activity

[0355] The binding specificity of the anti-WISP1 antibody against human WISP1 was evaluated using a direct coating ELISA assay (Fig. 2). Full-length human WISP1 protein (human_FL_CCN4) was diluted to a concentration of 5 μg / mL in 100 μl of 1x coating buffer (Biolegend cat#421701) per well and immobilized on Immulon-2HB plates (ThermoFischer Scientific cat#3445) at room temperature for 1 hour. The antigen solution was aspirated, washed three times with approximately 300 μl of 1 x TBST (VWR cat#K873), and then incubated with 200 μl of blocking solution at room temperature for 1 hour. The plates were subsequently washed three times with 1 x TBST, appropriate serial dilutions of the antigen were prepared, added to the plates, and incubated for 1 hour. After incubation with the primary antibody, the plate was washed as previously described, and 100 μl of anti-human IgG Fc gamma-HRP (Jackson Immunoresearch cat#109-035-170) was added at a 5000-fold dilution and incubated at room temperature for 1 hour. The plate was washed, 100 μl of TMB chromogenic reagent was added, and color development was allowed for 1 minute. After color development, 100 μl of TMB stop solution was added, and the absorbance was measured at 450 nm.

[0356] Binding kinetics were established by surface plasmon resonance (SPR) performed by Genscript USA (Fig. 3). Human full-length WISP1 was immobilized on a Series S sensor chip CM5 (GE Healthcare cat# BR-1005-30) by activating the sensor chip using 50 mM N-hydroxysuccinamide (NHS) and 200 mM 1-ethyl-3-93-dimethylaminopropyl carbodiimide hydrochloride (EDC). The WISP1 antigen was diluted to 30 mM in sodium acetate buffer, pH 4.5, and flowed onto the sensor chip to promote amine coupling reactions. After diluting WISP1 to appropriate concentrations, affinity measurements were performed in HBS-EP+ buffer at pH 7.4 at 25°C. Data were processed using Biacore 8K evaluation software version 1.1, and flow cytometry and blank injection buffer were used as dual reference materials.

[0357] Example 3: Analysis of WISP1 Domain Binding

[0358] The binding of anti-WISP1 antibodies to different domains of the WISP1 protein was evaluated by ELISA (Fig. 4). Human full-length WISP1 (Hu_FL_CCN4), domain 1 WISP1 (human CCN4-domain 1), WISP1 domains 1 and 2 (human CCN4-domain 12), WISP1 domains 3 and 4 (human CCN4-domain 34), and WISP1 domain 4 (human CCN4-domain 4) constructs were directly coated onto plates (Immulon-2HB plates; Thermo Scientific cat# 3455) at a concentration of 5 μg / mL for 1 hour at room temperature. The antigen solution was aspirated, washed twice with approximately 300 μl of 1 x TBST (VWR cat#K873), and then incubated with 200 μl of blocking solution at room temperature for 1 hour. The blocking solution was aspirated, and 100 μl of M6-NYS antibody and Ab162, expressed to have a mouse IgG2 constant region, were added to the wells at a concentration of 0.1 μg / mL. After incubation with the primary antibody, the plate was washed as previously described, 100 μl of anti-human IgG Fc gamma-HRP (Jackson Immunoresearch cat#109-035-170) was added at a 5000-fold dilution, and incubated at room temperature for 1 hour. The plate was washed, 100 μl of TMB chromogenic reagent was added, and color development was allowed for 1 minute. After color development, 100 µl of TMB stop solution was added, and the absorbance was measured at 450 nm. The K4 anti-WISP1 antibody was shown to bind to domain 2 of the WISP1 protein (Fig. 4).

[0359] Example 4: Carbon tetrachloride (CCL) in liver fibrosis 4 In vivo study demonstrating the therapeutic effect of anti-WISP1 antibodies in a mouse model

[0360] To induce liver fibrosis, male BALB / c mice were administered carbon tetrachloride (CCl4) in mineral oil at a 1:1 ratio (1 ml / kg) twice a week via the intraperitoneal route for 6 weeks. Therapeutic therapy was initiated on day 21 after the first CCl4 administration and continued until the end of the study on day 41. Animals were treated intraperitoneally with a vehicle or test antibody three times a week.

[0361] At the end of the study, mice were euthanized and liver tissue was collected. Fibrosis was measured by quantifying the collagen content in the liver using hydroxyproline.

[0362] CCL4 induced an increase in collagen content in the liver, as indicated by an increase in hydroxyproline production compared to the control group. Anti-WISP1 antibodies inhibited liver fibrosis. 3 mg / kg of M6-QYS antibody caused a 79.7% reduction in hydroxyproline compared to the vehicle group (+PBS). 10 mg / kg of anti-WISP1 antibody K4 caused a 76.4% reduction in hydroxyproline compared to the vehicle group (+PBS) (Figs. 5a-5b). The anti-WISP1 antibodies used in this study were expressed as having a mouse IgG constant region.

[0363] The results indicate that the anti-WISP1 antibody is effective in inhibiting liver fibrosis.

[0364] Example 5: In vivo study demonstrating the therapeutic effect of anti-WISP1 antibody in a mouse model of bile duct ligation (BDL) in liver fibrosis

[0365] Liver injury and fibrosis were induced in male Sprague-Dawley rats by performing sham surgery or bile duct ligation (BDL) on Day 0. Rats were administered 10 mg / kg of M6-QYS antibody or K4 antibody every 3 days (Q3D) from Day 3 to Day 18. The anti-WISP1 antibody was expressed with a mouse IgG2 constant region. All administrations were performed as ip at a volume equivalent to 2 mL / kg. Rats were humanely euthanized, and tissues were collected on Day 21. Effects on myofibroblasts were measured by alpha-smooth muscle actin (α-SMA) staining in the liver.

[0366] One section (5 μm) of paraffin-embedded liver tissue was obtained and mounted. The section was stained for α-smooth muscle actin (α-SMA) using a standard immunohistochemistry (IHC) method. Images of the liver tissue stained for α-SMA (n = 10 / section) were obtained using a Zeiss AxioImager.A2 microscope at 100x magnification (sufficient to obtain a drawing of ~60–70% of the area) and applied for quantitative image analysis. Quantitative histological image analysis of the anti-α-SMA-stained tissue sections was performed using the color spectral segmentation method. α-SMA (percentage of total tissue area imaged) appeared as an average positive diaminobenzidine (DAB) stain for the sampled images (Fig. 6a).

[0367] A representative optical microscope image of liver tissue stained for α-SMA is exemplified in Fig. 6b.

[0368] BDL increased α-SMA staining in the liver compared to the sham-operated control group (7.96 ± 0.35 vs. 0.01 ± 0.00%). M6-QYS and K4 antibodies attenuated the BDL-induced increase in α-SMA (M6-QYS: 5.33 ± 0.47; and K4: 4.84 ± 0.47 vs. vehicle: 7.96 ± 0.35%).

[0369] The results indicate that the anti-WISP1 antibody is effective in attenuating the BDL-induced increase in liver alpha-SMA.

[0370] Example 6: Anti-WISP1 antibody K4 inhibits pSMAD2

[0371] The conventional TGFβ1 signaling pathway involves the activation of SMAD (Small Mother Against Decapentaplegic) proteins through the phosphorylation of SMAD2 and SMAD3, which are major mediators of downstream TGFβ1-mediated fibrosis (Schmierer and Hall 2007; Frangogiannis 2020). Various reports have shown that WISP1 can act in relation to TGFβ signaling and, accordingly, exhibit a potential mechanism by which WISP1 can promote or exacerbate fibrosis (Wang et al., 2020; Zhang et al. 2020; Inkson et al. 2008).

[0372] Accordingly, the applicant evaluated the ability of anti-WISP1 antibodies to modulate pSMAD2 and pSMAD3 signaling. Normal rat renal fibroblasts (NRKF) were treated with recombinant human WISP1 (10 μg / mL) for 30 minutes in the absence or presence of anti-WISP1 antibodies, and pSMAD2 was evaluated in cell lysates by the Luminex assay. The ability of various antibodies to inhibit pSMAD2 signaling was determined relative to treatment with a homologous control antibody (inhibition %). The percentage of pSMAD2 inhibition ranged from 0% to 71% (Fig. 7). Monoclonal antibody E8 reduced pSMAD2 by 44%, while antibody K4 reduced the signal by 63%, which was similar to the inhibition by the polyclonal anti-WISP1 antibody AF1627 (65%; positive control). Similar results were observed for pSMAD3 (data not shown).

[0373] Example 7: Anti-WISP1 antibody K4 inhibits primary human hepatic astrocyte chemotaxis / motility.

[0374] Wound healing requires the localization of desired cells and factors to the site of injury to initiate repair. If this process is prolonged or disrupted due to sustained injury, fibrosis develops, leading to tissue stiffness and ultimately organ failure (Knoedler et al. 2023). In the case of the liver, differentiation of hepatic stellate cells into myofibroblasts has been shown to be a major factor in hepatic fibrosis (Tsuchida and Friedman, 2017). Biochemically, WISP1 has been shown to be able to linearize collagen I, a key component of the ECM. This activity has been shown to affect cell motility in vitro as well as cancer metastasis in vivo (Jia et al. EMBO 2019; Janajanam et al. 2021). Furthermore, the literature [Xi et al. (2022)] indicates that WISP1 is elevated in the plasma of NASH patients and that WISP1 increases the motility of HHSCs through integrin interactions.

[0375] To evaluate the ability of the monoclonal antibody K4 to modulate this phenotype, the upper chamber of a 96-well transwell plate was coated with recombinant human WISP1, and 10% FBS was provided to the lower chamber as a chemotactic. Primary HHSCs were added to the upper chamber at 1 or 2 K per well. Antibodies were added to each well at the indicated concentrations, and cell migration to the lower chamber was measured (shown at 48h). Without antibodies (0 μg / mL), significant chemotaxis occurred as an increase in cells on the lower side of the transwell (Fig. 8, first column). Treatment with IgG antibodies did not alter the chemotaxis rate compared to wells without antibodies, even at the highest concentration evaluated (Fig. 8, top column). Treatment with the polyclonal anti-WISP1 antibody (R&D AF1627), starting at 2 μg / mL, significantly reduced WISP1-induced chemotaxis (Fig. 8, middle column). Monoclonal anti-WISP1 antibody K4 also began to inhibit chemotaxis at 2 μg / mL, and inhibition increased at higher doses (Fig. 8, bottom column).

[0376] Quantification of HHSC chemotaxis / migration was calculated at the 48-hour mark. The "phase difference bottom area normalized to the initial peak" metric was used to calculate chemotaxis. This method takes the masked cell area on the basal plane (bottom) of the membrane and normalizes it to the masked area on the apical plane (top) of the membrane from the first scan (4 hours), representing the number of cells that migrated from the apical plane of the transwell to the basal plane. It was found that using the 4-hour mark as the starting point resulted in a slightly lower chemotaxis rate, but similarly, it resulted in more uniform values ​​across the plate due to the significant morphological changes that occur immediately after seeding and during the first few hours while cells attach to the plate.

[0377] Significant HHSC chemotaxis occurred with the combination of WISP1 and 10% fetal bovine serum (FBS), but not in the absence of fetal bovine serum (FBS) (Fig. 9). Non-specific IgG antibodies did not inhibit chemotaxis / cell migration even at the peak concentration (100 μg / mL). The polyclonal anti-WISP1 antibody AF1627 inhibited chemotaxis starting at 2 μg / mL, and inhibition increased with higher antibody concentrations. The monoclonal anti-WISP1 antibody K4 significantly inhibited chemotaxis starting at 2 μg / mL, and inhibition increased with antibody concentration. Significance is shown against 100 μg / mL IgG with 10% FBS (* p < 0.05).

[0378] IC 50 and IC 80 The values ​​were determined using a Python-based nonlinear regression curve fit to the dose-response data from the final time point (48 hours; Fig. 10). The monoclonal anti-WISP1 antibody K4 had an IC50 of 1.8788 μg / mL. 50 and an IC50 of 4.7959 μg / mL 80 It generated; the polyclonal anti-WISP1 antibody AF1627 had an IC50 of 1.6629 μg / mL. 50 and an IC5 of 5.152 μg / mL 80 has

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[0385] References

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Claims

Claim 1 An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and the light chain variable domain comprises CDR L1 as presented in SEQ ID NO. 4, CDR L2 including AAS, and CDR L3 as presented in SEQ ID NO.

6. Claim 2 In claim 1, the antibody wherein the heavy chain variable domain comprises a sequence having at least 90% identity with respect to sequence number 13. Claim 3 In claim 1, the heavy chain variable domain is an antibody comprising a sequence having at least 95% identity with respect to sequence number 13. Claim 4 In claim 1, the antibody wherein the heavy chain variable domain comprises a sequence having at least 98% identity with respect to sequence number 13. Claim 5 In claim 1, the heavy chain variable domain is an antibody comprising sequence number 13. Claim 6 In claim 1, the light chain variable domain is an antibody comprising a sequence having at least 90% identity with respect to sequence number 14. Claim 7 In claim 1, the light chain variable domain is an antibody comprising a sequence having at least 95% identity with respect to sequence number 14. Claim 8 In claim 1, the light chain variable domain is an antibody comprising a sequence having at least 98% identity with respect to sequence number 14. Claim 9 In claim 1, the light chain variable domain is an antibody comprising SEQ ID NO.

14. Claim 10 In paragraph 1, an antibody that is IgG. Claim 11 In paragraph 1, an antibody that is a humanized antibody or a chimeric antibody. Claim 12 In claim 1, an antibody comprising a Fab fragment. Claim 13 In paragraph 1, the antibody is a single-chain antibody (scFv). Claim 14 An antibody capable of binding to the WISP1 protein in claim 1. Claim 15 In paragraph 14, the WISP1 protein is an antibody that is a human WISP1 protein. Claim 16 In claim 14, the WISP1 protein is an antibody comprising the sequence of sequence number 19 or 20. Claim 17 In claim 14, an antibody capable of binding to domain 2 of the WISP1 protein. Claim 18 In claim 14, the domain 2 of the WISP1 protein is an antibody comprising residues corresponding to 121 to 203 of sequence number 19. Claim 19 In paragraph 14, an equilibrium dissociation constant (K) of about 0.01 nM to about 20 nM D An antibody capable of binding to the above WISP1 protein. Claim 20 In claim 19, an antibody capable of binding to the WISP1 protein with a KD of about 5.55 nM. Claim 21 An anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody comprising: a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 comprising AAS, and CDR L3 as presented in SEQ ID NO. 6, which binds to the same epitope as the anti-WISP1 antibody. Claim 22 An antibody competing with an anti-WNT1-inducible-signaling pathway protein 1 (WISP1) antibody for binding to a WISP1 protein, wherein the anti-WISP1 antibody comprises a heavy chain variable domain comprising CDR H1 as presented in SEQ ID NO. 1, CDR H2 as presented in SEQ ID NO. 2, and CDR H3 as presented in SEQ ID NO. 3; and a light chain variable domain comprising CDR L1 as presented in SEQ ID NO. 4, CDR L2 comprising AAS, and CDR L3 as presented in SEQ ID NO.

6. Claim 23 In paragraph 1, an antibody attached to a therapeutic agent. Claim 24 In paragraph 1, the antibody attached to the diagnostic agent. Claim 25 A method for treating fibrosis in a subject requiring treatment for fibrosis, comprising the step of administering a therapeutically effective amount of the antibody of claim 1 to the subject. Claim 26 In paragraph 25, the above fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis. Claim 27 In paragraph 26, the above fibrosis is a fibrotic liver disease. Claim 28 In paragraph 26, the above fibrosis is pulmonary fibrosis. Claim 29 In paragraph 28, the subject has interstitial lung disease. Claim 30 In paragraph 29, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis. Claim 31 In paragraph 28, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis. Claim 32 In paragraph 31, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-related pulmonary fibrosis, or progressive pulmonary fibrosis. Claim 33 In paragraph 32, the pulmonary fibrosis is idiopathic pulmonary fibrosis. Claim 34 In paragraph 26, the above fibrosis is cardiac fibrosis. Claim 35 In paragraph 25, the subject has a method of having scleroderma. Claim 36 A method for treating fibrosis in a subject requiring treatment for fibrosis, comprising: a) confirming, in a biological sample obtained from the subject, an increased level of WISP1 compared to a standard control, an increased level of α-smooth muscle actin (α-SMA) compared to a standard control, an increased level of collagen type I alpha 1 chain (Col1a1) compared to a standard control, an increased level of tissue metalloproteinase inhibitor 1 (Timp1) compared to a standard control, an increased level of fibronectin (Fn1) compared to a standard control, an increased level of interleukin 6 (IL-6) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2) compared to a standard control, an increased level of phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3) compared to a standard control, or a combination thereof; and b) administering a therapeutically effective amount of the anti-WISP1 antibody of claim 1 to the subject. Claim 37 In paragraph 36, the above fibrosis is fibrotic liver disease, pulmonary fibrosis, or cardiac fibrosis. Claim 38 In paragraph 36, the above fibrosis is a fibrotic liver disease. Claim 39 In paragraph 36, the above fibrosis is pulmonary fibrosis. Claim 40 In paragraph 39, the subject has interstitial lung disease. Claim 41 In paragraph 40, the interstitial lung disease is non-specific interstitial pneumonia, rheumatoid arthritis interstitial lung disease, hypersensitivity pneumonia, beryllium, respiratory bronchiolitis interstitial lung disease, desquamative interstitial pneumonia, sarcoidosis, acute interstitial pneumonia, idiopathic organizing pneumonia, lymphocytic interstitial pneumonia, or pleural parenchymal elastic fibrosis. Claim 42 In paragraph 39, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-associated pulmonary fibrosis, progressive pulmonary fibrosis, Sjögren's syndrome-associated pulmonary fibrosis, pneumoconiosis, asbestosis, silicosis, radiation-induced pulmonary fibrosis, drug-induced pulmonary fibrosis, COVID-19-associated pulmonary fibrosis, pulmonary fibrosis after acute respiratory distress syndrome, lymphangioleiomyomatosis-associated pulmonary fibrosis, familial pulmonary fibrosis, Hermanski Pudlak syndrome-associated pulmonary fibrosis, or congenital dyskeratosis-associated pulmonary fibrosis. Claim 43 In paragraph 42, the pulmonary fibrosis is idiopathic pulmonary fibrosis, systemic sclerosis-related pulmonary fibrosis, or progressive pulmonary fibrosis. Claim 44 In paragraph 43, the pulmonary fibrosis is idiopathic pulmonary fibrosis. Claim 45 In paragraph 37, the above fibrosis is cardiac fibrosis. Claim 46 In paragraph 36, the subject has a method of having scleroderma. Claim 47 A method for inhibiting the release of interleukin 6 (IL-6), phosphorylated Small Mother Against Decapentaplegic 2 (pSMAD2), phosphorylated Small Mother Against Decapentaplegic 3 (pSMAD3), or a combination thereof from a cell, comprising the step of contacting the cell with the anti-WISP1 antibody of claim 1. Claim 48 In paragraph 47, the cell is an immune cell. Claim 49 In paragraph 47, the cell is a fibroblast. Claim 50 A method for inhibiting the migration of WISP-1 receptor-expressing cells, comprising the step of contacting the cells with the anti-WISP1 antibody of claim 1. Claim 51 In paragraph 50, the method wherein the WISP-1 receptor is an integrin. Claim 52 In paragraph 50, the cell is a fibroblast or an astrocyte. Claim 53 A detection method for WISP1-expressing cells comprising: (i) contacting WISP1-expressing cells with the antibody of claim 1; and (ii) detecting the binding of the antibody to a WISP1 protein expressed by the cells. Claim 54 In paragraph 53, the antibody is attached to a detectable moiety.

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