Novel function of CSF3 inhibitor and use thereof

Abstract

Disclosed herein is a therapeutic agent for fibrosis comprising a CSF3 inhibitor. A therapeutic agent for fibrosis comprising a CSF3R inhibitor is also disclosed herein. In addition, the specification discloses a novel anti-CSF3 antibody.

Description

Novel functions of CSF3 inhibitors and their applications

[0001] The present disclosure discloses a mechanism of action of a CSF3 inhibitor that causes fibrotic tissue to return to normal tissue; a mechanism of action of a CSF3R inhibitor that causes fibrotic tissue to return to normal tissue; a therapeutic agent and method for treating fibrosis and related diseases; and technology related to a CSF3 antibody.

[0002] Fibrosis is a condition characterized by excessive accumulation of extracellular matrix, most notably collagen, in tissues. When fibrosis occurs, tissues lose their original function and become rigid due to the accumulation of extracellular matrix. Fibrosis can occur in virtually any tissue in the body, and it is known that fibrosis in many tissues can lead to various diseases, disorders, and / or conditions. Fibrosis can be caused by persistent tissue irritation or severe tissue damage, and is also the final and common pathological consequence of many chronic inflammatory diseases.

[0003] Colony-Stimulating Factor 3 (CSF3), also known as Granulocyte colony-stimulating factor (G-CSF), is known to stimulate the proliferation, differentiation, aggregation, and terminal cell functional activation of granulocytes.

[0004] The purpose of this specification is to elucidate the fibrosis treatment mechanism related to CSF3 or CSF3R inhibition, and further to provide a fibrosis treatment method utilizing the same, a fibrosis treatment agent, uses of a substance related thereto, and antibodies related thereto.

[0005] In order to solve the above technical problem, the present specification provides the mechanism of action of the effect of a CSF3 inhibitor that returns fibrotic tissue to normal tissue; the mechanism of action of the effect of a CSF3R inhibitor that returns fibrotic tissue to normal tissue; and provides a method for treating fibrosis using the CSF3 inhibitor or the CSF3R inhibitor. The inventors of the present specification confirmed that the fibrosis treatment effect of a CSF3 inhibitor and a CSF3R inhibitor occurs regardless of the tissue in which fibrosis occurs, and therefore, can be an effective treatment mechanism for all types of fibrosis. The present specification proposes an anti-CSF3 antibody and a siRNA that suppresses the CSF3 gene as representative means for suppressing the CSF3, and their effectiveness was demonstrated through experiments. In addition, the present specification proposes an anti-CSF3R antibody and a siRNA that suppresses the CSF3R gene as representative means for suppressing the CSF3R, and their effectiveness was demonstrated through experiments.

[0006] The method for treating fibrosis using a CSF3 inhibitor or CSF3R inhibitor provided herein can prevent or treat fibrosis regardless of the type of tissue in which fibrosis occurs.

[0007] Figure 1 shows the results of analyzing the mRNA expression of α-SMA, Col1A, and Fibronectin using qPCR 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.1. Here, Cont is a control group treated with PBS, and rh-CSF3 50 ng / ml, rh-CSF3 100 ng / ml, and rh-CSF3 200 ng / ml are groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0008] Figure 2 shows the results of Western blot analysis of α-SMA, Col1A, and Fibronectin protein expression and Stat3 activation (p-Stat3) 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.1. Here, Cont is the control group treated with PBS, and 50, 100, and 200 under rh-CSF3 are groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0009] Figure 3 shows the results of analyzing α-SMA and Col1A proteins by immunofluorescence staining 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.1. Here, Cont is a control group treated with PBS, and 50 ng / ml, 100 ng / ml, and 200 ng / ml under rh-CSF3 are groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0010] Figure 4 shows the results of analyzing the secretion and accumulation of collagen using a hydroxyproline assay 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.1. Here, Cont is a control group treated with PBS, and rh-CSF3 50 ng / ml, rh-CSF3 100 ng / ml, and rh-CSF3 200 ng / ml are groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0011] Figure 5 shows the results of analyzing the mRNA expression of TGF-β using qPCR 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.2. Here, Control refers to the control group treated with PBS, and rh-CSF3 50 ng / ml, rh-CSF3 100 ng / ml, and rh-CSF3 200 ng / ml refer to the groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0012] Figure 6 shows the results of analyzing Smad3 activation (p-Smad3) by Western blot 24 hours after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.2. Here, Cont is the control group treated with PBS, and 50, 100, and 200 under rh-CSF3 are groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0013] Figure 7 shows the results of analyzing the concentration of TGF-β protein 24 hours later using a TGF-β ELISA assay after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein according to Experimental Example 2.2. Here, Control refers to the control group treated with PBS, and rh-CSF3 50 ng / ml, rh-CSF3 100 ng / ml, and rh-CSF3 200 ng / ml refer to the groups treated with CSF3 recombinant protein at concentrations of 50 ng / ml, 100 ng / ml, and 200 ng / ml, respectively.

[0014] Figure 8 shows the results of analyzing the mRNA expression of TGF-β and IL-11 using qPCR after treating lung fibroblasts (DHLF-IPF) with an antibody that does not target CSF3, an anti-CSF3 antibody, si-RNA that does not target the mRNA of CSF3 and CSF3R (hereinafter referred to as 'si-cont'), and si-RNA that targets the mRNA of CSF3 (hereinafter referred to as 'si-CSF3') according to Experimental Example 2.3. Here, IPF+IgG is a group treated with an antibody that does not target CSF3, IPF+CSF3 Ab is a group treated with an anti-CSF3 antibody, IPF+si-Cont is a group treated with si-cont, and IPF+si-CSF3 is a group treated with si-CSF3.

[0015] Figure 9 shows the results of analyzing the mRNA expression of TGF-β and IL-11 using qPCR after treating lung fibroblasts (BLM-MLF) isolated from bleomycin-induced pulmonary fibrosis mouse lung tissues with antibodies that do not target CSF3, anti-CSF3 antibodies, si-cont, and si-CSF3, respectively, according to Experimental Example 2.3. Here, MLF is a group of lung fibroblasts (MLF) isolated from normal mouse lung tissues, BLM-MLF +IgG is a group treated with an antibody that does not target CSF3, BLM-MLF +CSF3 Ab is a group treated with an anti-CSF3 antibody, BLM-MLF +si-Cont is a group treated with si-cont, and BLM-MLF +si-CSF3 is a group treated with si-CSF3.

[0016] Figure 10 shows the results of analyzing SMAD3 activation (p-SMAD3) using Western Blot after treating lung fibroblasts (DHLF-IPF) with antibodies that do not target CSF3, anti-CSF3 antibodies, si-cont, and si-CSF3, respectively, according to Experimental Example 2.3. Here, IgG is the group treated with antibodies that do not target CSF3, CSF3 Ab is the group treated with anti-CSF3 antibodies, si-C is the group treated with si-cont, and si-CSF3 is the group treated with si-CSF3.

[0017] Figure 11 shows the results of analyzing SMAD3 activation (p-SMAD3) through Western Blot after treating lung fibroblasts (BLM-MLF) isolated from bleomycin-induced pulmonary fibrosis mouse lung tissue with antibodies that do not target CSF3, si-cont, and si-CSF3, respectively, according to Experimental Example 2.3. Here, MLF is a group of lung fibroblasts (MLF) isolated from normal mouse lung tissue, IgG is a group treated with an antibody that does not target CSF3, CSF3 Ab is a group treated with an anti-CSF3 antibody, si-C is a group treated with si-cont, and si-CSF3 is a group treated with si-CSF3.

[0018] Figure 12 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, CSF3, CSF3R, TGF-β, and IL11 using qPCR after treating normal lung fibroblasts (MRC5) with si-cont, CSF3 recombinant protein, and si-RNA targeting the mRNA of CSF3R (hereinafter referred to as 'si-CSF3R') according to Experimental Example 2.4. Here, si-cont is a group treated with si-cont, si-cont+rhCSF3 is a group treated with si-cont and CSF3 recombinant protein, and si-CSF3R+rhCSF3 is a group treated with si-CSF3R and CSF3 recombinant protein.

[0019] Figure 13 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, CSF3, CSF3R, TGF-β, and IL11 using qPCR after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein and anti-CSF3R antibody, respectively, according to Experimental Example 2.4. Here, MRC5 is the control group, rh-CSF3 is the group treated with CSF3 recombinant protein, and rh-CSF3+CSF3R Ab is the group treated with CSF3 recombinant protein and anti-CSF3R antibody.

[0020] Figure 14 shows the results of analyzing the expression of α-SMA and Col1A1 proteins using Western Blot after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein and si-CSF3R, respectively, according to Experimental Example 2.4. Here, Cont is the control group, CSF3 is the group treated with CSF3 recombinant protein, and CSF3+si-CSF3R is the group treated with CSF3 recombinant protein and si-CSF3R.

[0021] Figure 15 shows the results of analyzing the expression of α-SMA and Col1A1 proteins by immunofluorescence staining after treating normal lung fibroblasts (MRC5) with CSF3 recombinant protein and si-CSF3R, respectively, according to Experimental Example 2.4. Here, Control is the control group, CSF3 is the group treated with CSF3 recombinant protein, and CSF3+si-CSF3R is the group treated with CSF3 recombinant protein and si-CSF3R.

[0022] Figure 16 shows the results of analyzing the TGF-β protein concentration using a TGF-β ELISA assay after treating normal lung fibroblasts (MRC5) with si-cont, CSF3 recombinant protein, and si-CSF3R, respectively, according to Experimental Example 2.4. Here, si-cont is a group treated with si-cont, si-cont+rhCSF3 is a group treated with si-cont and CSF3 recombinant protein, and si-CSF3R+rhCSF3 is a group treated with CSF3 recombinant protein and si-CSF3R.

[0023] Figure 17 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, CSF3, TGF-β, IL11, CSF3R, and TGF-βRII using qPCR 24 hours after treating normal lung fibroblasts (MRC5) with si-cont, CSF3 recombinant protein, si-CSF3R, and siRNA targeting the mRNA of TGF-β receptor 2 (TGF-βRII) (hereinafter referred to as 'si-TGF-βRII') according to Experimental Example 2.5. Here, si-cont is a group treated with si-cont, rh-CSF3 is a group treated with CSF3 recombinant protein, si-CSF3R+rh-CSF3 is a group treated with si-CSF3R and CSF3 recombinant protein, and si-TGF-B-RII+rhCSF3 is a group treated with si-TGF-βRII and CSF3 recombinant protein.

[0024] Figure 18 shows the results of analyzing the TGF-β protein concentration 24 hours later using a TGF-β ELISA assay after treating normal lung fibroblasts (MRC5) with si-cont, CSF3 recombinant protein, si-CSF3R, and si-TGF-βRII, respectively, according to Experimental Example 2.5. Here, Cont is the control group, rh-CSF3 is the group treated with CSF3 recombinant protein, si-CSF3R+rh-CSF3 is the group treated with si-CSF3R and CSF3 recombinant protein, and si-TGF-B-RII+rhCSF3 is the group treated with si-TGF-βRII and CSF3 recombinant protein.

[0025] Figure 19 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, CSF3, TGF-β, IL11, and CSF3R using qPCR 24 hours after treating normal lung fibroblasts (MRC5) with TGF-β, CSF3 antibody, and CSF3R antibody, respectively, according to Experimental Example 2.6. Here, Cont is the control group, TGFβ is the group treated with TGF-β, TGFβ+CSF3 Ab is the group treated with TGF-β and CSF3 antibody, and TGFβ+CSF3R Ab is the group treated with TGF-β and CSF3R antibody.

[0026] Figure 20 shows the results of analyzing the expression of α-SMA and Col1A1 proteins by immunofluorescence staining 24 hours after treating normal lung fibroblasts (MRC5) with TGF-β and CSF3 antibodies, respectively, according to Experimental Example 2.6. Here, rh-TGF-β1+IgG is a group treated with an antibody that does not bind to TGF-β and CSF3, and rh-TGF-β1+CSF3 Ab is a group treated with TGF-β and CSF3 antibodies.

[0027] Figure 21 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, and CSF3 using qPCR after treating DHLF-IPF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1. Here, IPF+si-Cont is a group treated with si-cont, IPF+si-CSF3 is a group treated with si-CSF3, IPF+IgG is a group treated with a non-CSF3-targeting antibody, and IPF+CSF3 Ab is a group treated with an anti-CSF3 antibody.

[0028] Figure 22 shows the results of analyzing the expression of α-SMA and Col1A1 proteins by immunofluorescence staining after treating DHLF-IPF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1.1. Here, IPF+si-Cont is a group treated with si-cont, IPF+si-CSF3 is a group treated with si-CSF3, IPF+IgG is a group treated with a non-CSF3-targeting antibody, and IPF+CSF3 Ab is a group treated with an anti-CSF3 antibody.

[0029] Figure 23 shows the results of analyzing the changes in the invasion ability of lung fibroblasts using a transwell cell culture chamber after treating DHLF-IPF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1.1. Here, IPF+si-Cont is the group treated with si-cont, IPF+si-CSF3 is the group treated with si-CSF3, IPF+IgG is the group treated with a non-CSF3-targeting antibody, and IPF+CSF3 Ab is the group treated with an anti-CSF3 antibody.

[0030] Figure 24 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, and CSF3 using qPCR after treating BLM-MLF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1.2. Here, BLM+si-Cont is a group treated with si-cont, BLM+si-CSF3 is a group treated with si-CSF3, BLM+IgG is a group treated with a non-CSF3-targeting antibody, and BLM+CSF3 Ab is a group treated with an anti-CSF3 antibody.

[0031] Figure 25 shows the results of analyzing the expression of α-SMA and Col1A1 proteins by immunofluorescence staining after treating BLM-MLF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1.2. Here, BLM+si-Cont is a group treated with si-cont, BLM+si-CSF3 is a group treated with si-CSF3, BLM+IgG is a group treated with a non-CSF3-targeting antibody, and BLM+CSF3 Ab is a group treated with an anti-CSF3 antibody.

[0032] Figure 26 shows the results of analyzing the changes in the invasion ability of lung fibroblasts using a transwell cell culture chamber after treating BLM-MLF cells with si-cont, si-CSF3, a non-CSF3-targeting antibody, and an anti-CSF3 antibody, respectively, according to Experimental Example 3.1.2. Here, BLM+si-Cont is the group treated with si-cont, BLM+si-CSF3 is the group treated with si-CSF3, BLM+IgG is the group treated with a non-CSF3-targeting antibody, and BLM+CSF3 Ab is the group treated with an anti-CSF3 antibody.

[0033] Figure 27 is a schematic diagram of the experimental model used to conduct the animal experiment of Experimental Example 3.2.

[0034] Figure 28 shows the results of analyzing the morphological changes in lung tissue detected from mice treated with PBS, bleomycin (BLM), and anti-CSF3 antibody according to Experimental Example 3.2.2 using H&E staining.

[0035] Figure 29 shows the results of analyzing collagen accumulation in lung tissue detected from mice treated with PBS, bleomycin, and anti-CSF3 antibody according to Experimental Example 3.2.2 using Massion trichrome staining.

[0036] In Figures 28 and 29, #1, #2, and #3 are lung tissues detected from different mouse individuals, PBS is a group treated with PBS, BLM is a group treated with bleomycin, and BLM+CSF3-4E5 is a group treated with bleomycin and anti-CSF3 antibody.

[0037] Figure 30 shows the results of analyzing the amount of collagen in lung tissue detected from mice treated with PBS, bleomycin, and anti-CSF3 antibody according to Experimental Example 3.2.3, using hydroxyproline content measurement.

[0038] Figure 31 shows the results of analyzing the mRNA expression of α-SMA and Col1A1 in lung tissues detected from mice treated with PBS, bleomycin, and anti-CSF3 antibody, respectively, according to Experimental Example 3.2.4, using qPCR.

[0039] Figure 32 shows the results of analyzing the secretion of TGF-β protein in lung tissue detected from mice treated with PBS, bleomycin, and anti-CSF3 antibody according to Experimental Example 3.2.4 using a TGF-β ELISA assay.

[0040] Figure 33 shows the results of analyzing the mRNA expression of TGF-β in lung tissue detected from mice treated with PBS, bleomycin, and anti-CSF3 antibody according to Experimental Example 3.2.4 using qPCR (Figure 33A) and analyzing the phosphorylation of SMAD3 protein (p-SMAD3) using Western Blot (Figure 33B).

[0041] In Figures 30 to 33, PBS is a group treated with PBS, BLM is a group treated with bleomycin, and BLM+CSF3-4E5 or BLM+4E5 is a group treated with bleomycin and anti-CSF3 antibody. Here, G1-1, G1-2, G1-3, G1-4, and G1-5 each belong to the PBS-treated group and are different mouse individuals, G2-1, G2-3, and G2-5 each belong to the bleomycin-treated group and are different mouse individuals, and G3-1, G3-2, and G3-3 each belong to the bleomycin-treated group and are different mouse individuals.

[0042] Figures 34, 35, and 36 show the expression levels of CSF3, α-SMA, Col1A1, TGF-β, CTGF, TNF-α, IL8, and IL11 measured in cells when dermal fibroblasts were treated with PBS, TGF-β, and / or CSF3 antibody according to Experimental Examples 3.3.1 and 3.3.2. The left side of Figure 34 shows the expression levels of α-SMA and Col1A1 proteins confirmed through western blot. The right side of Figure 34 shows the expression levels of α-SMA and Col1A1 proteins confirmed through fluorescent staining. Figure 35 shows the expression levels of mRNA encoding CSF3, α-SMA, and Col1A1 proteins confirmed through q-PCR. Figure 36 shows the expression levels of mRNA of TGF-β, CTGF, TNF-α, IL8, and IL11 confirmed through q-PCR. At this time, in FIGS. 34, 35, and 36, Cont means a group treated only with PBS, TGF-β means a group treated only with TGF-β, and TGF-β+CSF3 Ab means a group treated with TGF-β and CSF3 antibody.

[0043] Figure 37 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, TGF-β, IL11, CSF3, and CSF3R using q-PCR after skin fibroblasts were treated with TGF-β and anti-CSF3R antibody, respectively, according to Experimental Example 3.3.3. Here, TGF-β is a group treated with TGF-β, and TGF-β+CSF3R Ab is a group treated with TGF-β and anti-CSF3R antibody.

[0044] Figure 38 shows the results of analyzing the expression of CSF3 protein through Western Blot after bleomycin, si-cont, and si-CSF3 were each treated on skin fibroblasts according to Experimental Example 3.3.4.

[0045] Figure 39 shows the results of analyzing the mRNA expression of CSF3, α -SMA, Col1A1, and TGF-β through qPCR after skin fibroblasts were treated with bleomycin, si-cont, and si-CSF3, respectively, according to Experimental Example 3.3.4.

[0046] In Figures 38 and 39, si-Cont is a group treated with si-cont, si-cont_BLM is a group treated with bleomycin and si-cont, and si-CSF3-#1+BLM, si-CSF3-#2+BLM, and si-CSF3-#3+BLM are groups treated with si-CSF3 and bleomycin.

[0047] Figure 40 is a schematic diagram of the experimental model used to conduct the animal experiment of Experimental Example 3.4.

[0048] Figures 41, 42, and 43 show skin tissues observed in a control group treated with only PBS, an experimental group treated with only Bleomycin, and an experimental group treated with Bleomycin and CSF3 antibody according to Experimental Example 3.4.2 in C57BL / 6 mice. Figure 41 shows the results of H&E staining of mouse skin tissue. Figure 42 shows the results of massion trichrime staining of collagen in mouse skin tissue. Figure 43 shows the results of α-SMA and Col1A1 protein staining in mouse skin tissue. In this case, in Figures 41 to 43, Cont refers to a group treated with only PBS, BLM refers to a group treated with only Bleomycin, and BLM+CSF3 Ab refers to a group treated with bleomycin and CSF3 antibody.

[0049] Figure 44 is a schematic diagram of the experimental model used to conduct the animal experiment of Experimental Example 3.4.

[0050] Figure 45 shows the hair loss phenomenon and hair loss recovery phenomenon observed in the control group administered only PBS to mice, the experimental group administered only Bleomycin, and the experimental group administered CSF3 antibody in addition to Bleomycin according to Experimental Example 3.4.3. In this case, in Figure 45, PBS refers to the group treated only with PBS, BLM refers to the group treated only with Bleomycin, and BLM+CSF3 Ab refers to the group treated with Bleomycin and CSF3 antibody.

[0051] Figure 46 is a schematic diagram of the experimental model used to conduct the experiment of Experimental Example 3.4.4.

[0052] Figure 47 shows the expression levels of CSF3, α-SMA, Col1A1, and TGF-β observed in a control group treated with only PBS, an experimental group treated with only Bleomycin, and an experimental group treated with CSF3 antibody in addition to Bleomycin according to Experimental Example 3.4.4. Figure 47 shows the expression levels of the genes CSF3, α-SMA, Col1A1, and TGF-β. In this case, in Figure 47, Cont refers to a group treated with only PBS, BLM refers to a group treated with only Bleomycin, and BLM+CSF3 Ab refers to a group treated with both bleomycin and CSF3 antibody.

[0053] Figures 48 to 52 relate to the expression levels of CSF3, α-SMA, Col1A1, and TGF-β in the tissues of two burn patients according to Experimental Example 3.4.5. P1-N refers to the normal tissue (or fibroblasts in the normal tissue) of the first patient. P1-F refers to the hypertrophic scar tissue (or fibroblasts in the hypertrophic scar tissue) of the first patient. P2-N refers to the normal tissue (or fibroblasts in the normal tissue) of the second patient. P2-F refers to the hypertrophic scar tissue (or fibroblasts in the hypertrophic scar tissue) of the second patient.

[0054] Figure 53 shows the results of analyzing the mRNA expression of -SMA, Col1A1, FN, CSF3, CSF3R, TGF-β, and IL11 using qPCR after human hepatic stellate cell line (LX-2 cells) was treated with PBS, TGF-β, anti-CSF3 antibody, and anti-CSF3R antibody, respectively, according to Experimental Example 3.5. Here, LX2 is a group treated with PBS, TGF-β is a group treated with TGF-β, TGF-β+CSF3 Ab is a group treated with TGF-β and anti-CSF3 antibody, and TGF-β+CSF3R Ab is a group treated with TGF-β and anti-CSF3R antibody.

[0055] Figure 54 shows the results of analyzing the mRNA expression of -SMA, Col1A1, FN, CSF3, CSF3R, TGF-β, and IL11 using qPCR after human normal renal tubular epithelial cells (HK-2 cells) were treated with PBS, TGF-β, anti-CSF3 antibody, and anti-CSF3R antibody, respectively, according to Experimental Example 3.6. Here, HK2 is a group treated with PBS, TGF-β is a group treated with TGF-β, TGF-β+CSF3 Ab is a group treated with TGF-β and anti-CSF3 antibody, and TGF-β+CSF3R Ab is a group treated with TGF-β and anti-CSF3R antibody.

[0056] Figures 55 to 57 show the results of analyzing the binding of candidate antibodies to the CSF3 protein through ELISA according to Experimental Example 5.2.1. Here, Figure 57 is a graph representing the results of Figures 55 and 56.

[0057] Figures 58 to 63 show the results of analyzing the neutralization ability of candidate antibodies against CSF3 through ELISA according to Experimental Example 5.2.2. Here, Figures 59 to 63 are graphs representing the results of Figure 58. Figure 63 is a graph extracted only when the concentration of the candidate antibody in Figure 58 is 25 nM.

[0058] Figures 64 to 80 show the results of analyzing the purity of candidate antibodies using SEC-HPLC according to Experimental Example 5.2.3. Here, the mass of the candidate antibodies is approximately 158 (kDa).

[0059] Figure 81 shows the results of analyzing the melting points of candidate antibodies using the Protein Thermal Shift Dye Kit according to Experimental Example 5.2.4.

[0060] Figure 82 shows the results of analyzing the amount of collagen using the Hydroxyproline assay after human lung cells were treated with bleomycin and candidate antibodies, respectively, according to Experimental Example 5.2.5. Here, BLM is the group treated only with bleomycin, and Y33 to Y50 are groups treated with bleomycin and each candidate antibody.

[0061] Figures 83 to 86 show the results of analyzing the mRNA expression of α-SMA, Col1A1, TGF-β, and IL-11 using qPCR after human lung cells were treated with bleomycin and candidate antibodies, respectively, according to Experimental Example 5.2.6. Here, BLM is a group treated only with bleomycin, and Y33 to Y50 are groups treated with bleomycin and each candidate antibody.

[0062] Figure 87 shows the results of analyzing the expression of α-SMA and Col1A1 proteins by Western Blot after human lung cells were treated with bleomycin and candidate antibodies, respectively, according to Experimental Example 5.2.7. Here, BLM is a group treated only with bleomycin, IgG is a group treated with bleomycin and an antibody that does not bind to CSF3, REF is a group treated with bleomycin and an existing commercial antibody, and Y-34, Y-40, Y-41, and Y-47 are groups treated with bleomycin and each of the candidate antibodies.

[0063] Figure 88 shows the results of analyzing the expression of α-SMA and Col1A1 proteins through immunofluorescence staining after human lung cells were treated with bleomycin and candidate antibodies, respectively, according to Experimental Example 5.2.7. Here, BLM is a group treated only with bleomycin, BLM+IgG is a group treated with bleomycin and an antibody that does not bind to CSF3 (control antibody), BLM+REF is a group treated with bleomycin and an existing commercialized antibody, and BLM+Y-34, BLM+Y-40, BLM+Y-41, and BLM+Y-47 are groups treated with bleomycin and each candidate antibody.

[0064] Figure 89 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, CSF3, TGF-β, and IL11 using qPCR 24 hours after lung fibroblasts were treated with CSF3 recombinant protein and si-RNA targeting STAT3 mRNA (hereinafter referred to as 'si-STAT3') according to Experimental Example 2.7. Here, rh-CSF3 is a group treated with CSF3 recombinant protein, and rh-CSF3+STAT3-I is a group treated with CSF3 recombinant protein and si-STAT3.

[0065] Figure 90 shows the results of analyzing the expression of α-SMA, Col1A1, and FN proteins using Western Blot 24 hours after lung fibroblasts were treated with CSF3 recombinant protein and si-STAT3, respectively, according to Experimental Example 2.7. Here, rh-CSF3 is the group treated with CSF3 recombinant protein, and rh-CSF3+STAT3-I is the group treated with CSF3 recombinant protein and si-STAT3.

[0066] Figure 91 shows the results of analyzing the mRNA expression of α-SMA, Col1A1, FN, CSF3, TGF-β, IL11, and CSF3R using qPCR 24 hours after lung fibroblasts were treated with TGF-β, si-cont, and si-CSF3, respectively, according to Experimental Example 2.6. Here, si-cont is a group treated only with si-cont, TGF-β+si-Cont is a group treated with TGF-β and si-cont, TGF-β+si-CSF3#1 is a group treated with TGF-β and si-CSF3, and TGF-β+si-CSF3#2 is a group treated with TGF-β and si-CSF3.

[0067] Hereinafter, the best mode for carrying out the invention is exemplified. This includes some, but not all, implementations of the invention disclosed herein. The embodiments described in this paragraph are merely exemplary, and the implementations described in this paragraph should not be construed as the "best mode for carrying out the invention." Those skilled in the art will likely envision numerous variations and more desirable implementations of the examples described in this paragraph, and such variations should also be considered to be included within the best mode for carrying out the invention.

[0068] In the present specification, a CSF3 inhibitor for use in treating fibrosis is provided.

[0069] In one embodiment, the fibrosis may be selected from the following:

[0070] Hypertrophic scar; systemic sclerosis; multiple cancers; pulmonary arterial hypertension; glial scar; Alzheimer's disease; cardiac fibrosis; hypertrophic cardiomyopathy; cardiac dysfunction; valvular disease; arrhythmia; myelofibrosis; myelodysplastic syndrome; chronic myelogenous leukemia; cirrhosis; portal hypertension; hepatocellular carcinoma; nonalcoholic steatohepatitis (NASH); intestinal fibrosis; enteropathies; inflammatory bowel disease; Arthrofibrosis; subretinal fibrosis; epithelial fibrosis; vision loss; idiopathic pulmonary fibrosis; cystic fibrosis; pulmonary hypertension; thromboembolic disease; emphysema; renal fibrosis; cystic fibrosis; nephrogenic systemic fibrosis; chronic kidney disease;Renal anemia; retroperitoneal fibrosis; mediastinal fibrosis; pancreatic fibrosis; cystic fibrosis; chronic pancreatitis; duct obstruction; autoimmune-interstitial lung disease, including ankylosing spondylitis and rheumatoid arthritis; connective tissue disease-interstitial lung disease, including rheumatoid arthritis-interstitial lung disease; and non-idiopathic pulmonary fibrosis-interstitial lung disease.

[0071] In one embodiment, the CSF3 inhibitor can induce degradation of extracellular matrix accumulated within the tissue where the fibrosis has occurred.

[0072] In one example, the extracellular matrix accumulated within the tissue where the fibrosis has occurred may be collagen.

[0073] In one embodiment, the CSF3 inhibitor can inactivate myofibroblasts within the tissue where the fibrosis has occurred.

[0074] In one embodiment, inactivating myofibroblasts within the tissue in which the fibrosis has occurred may mean at least one selected from the following:

[0075] (a) decomposing the extracellular matrix secreted by the myofibroblasts;

[0076] (b) inhibiting extracellular matrix secretion of the above myofibroblasts; and

[0077] (c) Reducing the number of myofibroblasts within the tissue where the fibrosis has occurred.

[0078] In one embodiment, the extracellular matrix of (a) and (b) may be collagen.

[0079] In one embodiment, the above (b) may induce a decrease in the expression level of the COL1A1 gene in the tissue where the fibrosis has occurred.

[0080] In one embodiment, the above (c) may induce a decrease in the expression level of the α-SMA gene in the tissue where the fibrosis has occurred.

[0081] In one embodiment, (c) may occur when the myofibroblasts revert to epithelial cells.

[0082] The present disclosure provides a method for preventing or treating fibrosis, comprising:

[0083] Administering to a subject a therapeutically effective amount of a CSF3 inhibitor of any one of the above embodiments.

[0084] The present disclosure provides a pharmaceutical composition for treating fibrosis, comprising:

[0085] A therapeutically effective amount of a CSF3 inhibitor of any one of the above embodiments; and

[0086] Pharmaceutically acceptable carrier.

[0087] In one embodiment, the pharmaceutically acceptable carrier may be at least one selected from the following:

[0088] Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; sweeteners; flavoring agents; syrups; liquid carriers such as fatty oils; sterile aqueous solutions; injectable esters such as propylene glycol; polyethylene glycol; ethyl oleate; suspending agents; emulsions; lyophilized preparations; external preparations; stabilizers; buffers; animal oils; vegetable oils; waxes; paraffin; starches; tragacanth; cellulose derivatives; polyethylene glycols; silicones; bentonites; silica; talc; zinc oxide; and suitable combinations thereof.

[0089] The present specification provides a use of any one of the above examples for the preparation of a CSF3 inhibitor for the treatment of fibrosis.

[0090] The present specification provides the sequences of novel anti-CSF3 antibodies.

[0091] In the present specification, a CSF3R inhibitor for use in treating fibrosis is provided.

[0092] In one embodiment, the fibrosis may be selected from the following:

[0093] Hypertrophic scar; systemic sclerosis; multiple cancers; pulmonary arterial hypertension; glial scar; Alzheimer's disease; cardiac fibrosis; hypertrophic cardiomyopathy; cardiac dysfunction; valvular disease; arrhythmia; myelofibrosis; myelodysplastic syndrome; chronic myelogenous leukemia; cirrhosis; portal hypertension; hepatocellular carcinoma; nonalcoholic steatohepatitis (NASH); intestinal fibrosis; enteropathies; inflammatory bowel disease; Arthrofibrosis; subretinal fibrosis; epithelial fibrosis; vision loss; idiopathic pulmonary fibrosis; cystic fibrosis; pulmonary hypertension; thromboembolic disease; emphysema; renal fibrosis; cystic fibrosis; nephrogenic systemic fibrosis; chronic kidney disease;Renal anemia; retroperitoneal fibrosis; mediastinal fibrosis; pancreatic fibrosis; cystic fibrosis; chronic pancreatitis; duct obstruction; autoimmune-interstitial lung disease, including ankylosing spondylitis and rheumatoid arthritis; connective tissue disease-interstitial lung disease, including rheumatoid arthritis-interstitial lung disease; and non-idiopathic pulmonary fibrosis-interstitial lung disease.

[0094] In one embodiment, the CSF3R inhibitor can induce degradation of extracellular matrix accumulated within the tissue where fibrosis has occurred.

[0095] In one example, the extracellular matrix accumulated within the tissue where the fibrosis has occurred may be collagen.

[0096] In one embodiment, the CSF3R inhibitor can inactivate myofibroblasts within the tissue where the fibrosis has occurred.

[0097] In one embodiment, inactivating myofibroblasts within the tissue in which the fibrosis has occurred may mean at least one selected from the following:

[0098] (a) decomposing the extracellular matrix secreted by the myofibroblasts;

[0099] (b) inhibiting extracellular matrix secretion of the above myofibroblasts; and

[0100] (c) Reducing the number of myofibroblasts within the tissue where the fibrosis has occurred.

[0101] In one embodiment, the extracellular matrix of (a) and (b) may be collagen.

[0102] In one embodiment, the above (b) may induce a decrease in the expression level of the COL1A1 gene in the tissue where the fibrosis has occurred.

[0103] In one embodiment, the above (c) may induce a decrease in the expression level of the α-SMA gene in the tissue where the fibrosis has occurred.

[0104] In one embodiment, (c) may occur when the myofibroblasts revert to epithelial cells.

[0105] The present disclosure provides a method for preventing or treating fibrosis, comprising:

[0106] Administering to a subject a therapeutically effective amount of a CSF3R inhibitor of any one of the above embodiments.

[0107] The present disclosure provides a pharmaceutical composition for treating fibrosis, comprising:

[0108] A therapeutically effective amount of a CSF3R inhibitor of any one of the above embodiments; and

[0109] Pharmaceutically acceptable carrier.

[0110] In one embodiment, the pharmaceutically acceptable carrier may be at least one selected from the following:

[0111] Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; sweeteners; flavoring agents; syrups; liquid carriers such as fatty oils; sterile aqueous solutions; injectable esters such as propylene glycol; polyethylene glycol; ethyl oleate; suspending agents; emulsions; lyophilized preparations; external preparations; stabilizers; buffers; animal oils; vegetable oils; waxes; paraffin; starches; tragacanth; cellulose derivatives; polyethylene glycols; silicones; bentonites; silica; talc; zinc oxide; and suitable combinations thereof.

[0112] The present specification provides a use of any one of the above examples for the preparation of a fibrosis treatment agent.

[0113] Hereinafter, the present invention will be described in more detail through specific implementations and examples with reference to the attached drawings. It should be noted that the attached drawings include some, but not all, implementations of the invention. The invention disclosed by this specification may be implemented in various ways and is not limited to the specific implementations described herein. These implementations should be considered as provided to satisfy the legal requirements applicable to this specification. Those skilled in the art will be able to think of many modifications and other implementations of the invention disclosed herein. Therefore, the invention disclosed herein is not limited to the specific implementations described herein, and it should be understood that modifications and other implementations thereof are also included within the scope of the claims.

[0114] Definition of terms

[0115] The definitions of terms used in this disclosure are as follows.

[0116] approximately

[0117] The term "about" as used herein means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% relative to a reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

[0118] Amino acid sequence notation

[0119] Unless otherwise stated, when describing amino acid sequences in this disclosure, amino acid single-letter notation or three-letter notation is used, and is written in the N-terminal to C-terminal direction. For example, when written as RNVP, it means a peptide in which arginine, asparagine, valine, and proline are sequentially connected from the N-terminal to the C-terminal. As another example, when written as Thr-Leu-Lys, it means a peptide in which threonine, leucine, and lysine are sequentially connected from the N-terminal to the C-terminal. In the case of amino acids that cannot be expressed in the single-letter notation, other letters are used and additional explanations are provided.

[0120] The notation for each amino acid is as follows: Alanine (Ala, A); Arginine (Arg, R); Asparagine (Asn, N); Aspartic acid (Asp, D); Cysteine ​​(Cys, C); Glutamic acid (Glu, E); Glutamine (Gln, Q); Glycine (Gly, G); Histidine (His, H); Isoleucine (Ile, I); Leucine (Leu, L); Lysine (Lys, K); Methionine (Met, M); Phenylalanine (Phe, F); Proline (Pro, P); Serine (Ser, S); Threonine (Thr, T); Tryptophan (Trp, W); Tyrosine (Ty, Y); and Valine (Val, V).

[0121] Nucleic acid sequence notation

[0122] The symbols A, T, C, G, and U used in this disclosure are to be interpreted as meanings understood by a person skilled in the art. They may be appropriately interpreted as bases, nucleosides, or nucleotides on DNA or RNA, depending on the context and technology. For example, when referring to a base, they may be interpreted as adenine (A), thymine (T), cytosine (C), guanine (G), or uracil (U) themselves, respectively; when referring to a nucleoside, they may be interpreted as adenosine (A), thymidine (T), cytidine (C), guanosine (G), or uridine (U), respectively; and when referring to a nucleotide in a sequence, they should be interpreted as meaning a nucleotide containing each of the above nucleosides.

[0123] In the present disclosure, the symbol N may be appropriately interpreted as a base, a nucleoside, or a nucleotide on DNA or RNA, depending on the context and technology. For example, when referring to a base, it may be interpreted as any one of adenine (A), thymine (T), cytosine (C), guanine (G), and uracil (U), respectively; when referring to a nucleoside, it may be interpreted as any one of adenosine (A), thymidine (T), cytidine (C), guanosine (G), and uridine (U), respectively; and when referring to a nucleotide in a sequence, it should be interpreted as meaning a nucleotide including each of the above nucleosides.

[0124] vector

[0125] As used herein, "vector" refers to any material capable of transporting genetic material into a cell, unless otherwise specified. For example, a vector may contain the target genetic material in the form of a nucleic acid sequence. For example, it may be a DNA molecule containing a nucleic acid encoding a protein (e.g., an antibody), but is not limited thereto. The term encompasses all meanings recognizable to those skilled in the art and may be appropriately interpreted according to the context.

[0126] antibodies

[0127] generalization

[0128] The antibodies disclosed in this disclosure are described.

[0129] The structure of the above antibody is divided into a heavy chain region and a light chain region depending on the chain type. The structure of the above antibody is divided into a fragment antigen-binding region (Fab region) and a fragment crystallizable region (Fc region) depending on the function of binding to the antigen. The structure of the above antibody is divided into a variable region and a constant region depending on the variability of the amino acid sequence. Other structures of the above antibody include the hinge region and the tail region.

[0130] Structure of antibodies 1- Heavy and light chain regions

[0131] The antibody is structurally divided into a heavy chain region and a light chain region. The heavy chain region has a polypeptide chain structure and includes four or five immunoglobulin domains. The light chain region has a polypeptide chain structure and includes two immunoglobulin domains. Each immunoglobulin domain includes about 100 to 120 amino acids.

[0132] Antibody Structure 2- Fragment Antigen-Binding and Crystallizable Region

[0133] The heavy chain region and the light chain region are functionally largely divided into a fragment antigen-binding region (Fab region) and a fragment crystallizable region (Fc region). The Fab region is a region that includes a portion that binds to an antigen (antigen-binding site). The Fc region is a portion that can bind to an Fc receptor. The heavy chain region has both a Fab region and an Fc region. The light chain region has a Fab region.

[0134] Antibody Structure 3- Variable and Constant Regions

[0135] The Fab region of the heavy chain region includes a heavy chain variable region (VH) and a heavy chain constant region 1 (CH1). The Fc region of the heavy chain region includes a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3), or includes CH2, CH3, and a heavy chain constant region 4 (CH4). In this case, the entire heavy chain constant region of the antibody is referred to as CH. For example, the entire region combining CH1, CH2, and CH3 of IgG1 can be expressed as CH.

[0136] The Fab region of the light chain region includes a light chain variable region (VL) and a light chain constant region (CL). The light chain region does not have an Fc region. Each of the VH, CH1, CH2, CH3, CH4, VL, and CL is an immunoglobulin domain, and one immunoglobulin domain includes about 100 to 120 amino acids.

[0137] Antibody Structure 4- Linkage Relationships

[0138] The immunoglobulin domains included in the heavy chain region are positioned in the order of VH, CH1, CH2, and CH3 or VH, CH1, CH2, CH3, and CH4 from the N-terminal to the C-terminal. The immunoglobulin domains included in the light chain region are positioned in the order of VL and CL from the N-terminal to the C-terminal.

[0139] The heavy chain region and the light chain region are connected by a disulfide bond. Specifically, the disulfide bond between CH1 of the heavy chain region and CL of the light chain region is connected. The Fab region and the Fc region are connected by a hinge region. Specifically, the C-terminal portion of CH1 of the heavy chain region and the N-terminal portion of CH2 are connected by the hinge region. The hinge region is composed of about 10 to 70 amino acids.

[0140] Characteristics of variable areas

[0141] The variable regions (VH and VL) above are regions that include an antigen-binding portion. Therefore, unlike the constant regions (CH1, CH2, CH3, CH4, and CL), the variable regions may have different sequences depending on the type of antigen to which they bind and / or the binding site (epitope).

[0142] Even within the above variable region, there is a hypervariable region with the greatest variability, and this region is called the complementarity-determining region (CDR). One CDR contains approximately 3 to 15 amino acids.

[0143] The above VH contains three CDRs, and the three CDRs included in the VH are called CDRH1, CDRH2, or CDRH3, respectively. The CDRs in the VH are located in the order of CDRH1, CDRH2, and CDRH3 from the N-terminal to the C-terminal.

[0144] The above VL contains three CDRs, and the three CDRs included in the VL are referred to as CDRL1, CDRL2, or CDRL3, respectively. The CDRs in the VH are located in the order of CDRL1, CDRL2, and CDRL3 from the N-terminal to the C-terminal.

[0145] The portion of the variable region excluding the CDR is called the framework region (FR). The VH includes four FRs, and the four FRs included in the VH are respectively called FRH1, FRH2, FRH3, or FRH4. The FRs in the VH are located in the order of FRH1, FRH2, FRH3, and FRH4 from the N-terminal to the C-terminal.

[0146] The above VL includes four FRs, and the four FRs included in the VL are referred to as FRL1, FRL2, FRL3, or FRL4, respectively. The FRs in the VL are located in the order of FRL1, FRL2, FRL3, and FRL4 from the N-terminal to the C-terminal.

[0147] The VHs are located in the order of FRH1, CDRH1, FRH2, CDRH2, FRH3, CDRH3, and FRH4 from the N-terminal to the C-terminal. The VLs are located in the order of FRL1, CDRL1, FRL2, CDRL2, FRL3, CDRL3, and FRL4 from the N-terminal to the C-terminal.

[0148] Characteristics of the invariant region

[0149] The constant region of the antibody is a region separated from the antigen-binding portion. The constant region can interact with cells or molecules of the immune system. In one specific example, the constant region can interact with (be bound to or linked to) the cell membrane of an immune cell (e.g., a lymphocyte, neutrophil, dendritic cell, or macrophage). Specifically, the hinge region and / or CH2 portion of the constant region can bind to a receptor (e.g., Fc εRIII) on the cell membrane of the immune cell. In another specific example, the constant region can bind to FcRn.

[0150] The constant region of the heavy chain region (hereinafter referred to as “heavy chain constant region”) is broadly divided into five types (classes or isotypes): alpha (α), gamma (γ), delta (δ), epsilon (ε), and mu (μ). At this time, the type of the heavy chain constant region is not determined individually for CH1, CH2, CH3, and CH4, but is determined by considering all heavy chain constant regions (CH1, CH2, and CH3; or CH1, CH2, CH3, and CH4) included in the antibody.

[0151] There are two types of constant regions of the above light chain region (hereinafter referred to as “light chain constant regions”), namely lambda (λ) and kappa (κ).

[0152] Types of Antibodies - Overview

[0153] Antibodies are broadly divided into five types (classes or isotypes). These five types are determined by the type of heavy chain constant region.

[0154] There are five types of antibodies: Immunoglobulin M (IgM), Immunoglobulin D (IgD), Immunoglobulin G (IgG), Immunoglobulin A (IgA), and Immunoglobulin E (IgE). If the type of heavy chain constant region of the antibody is alpha, the antibody type is IgA. If the type of heavy chain constant region of the antibody is gamma, the antibody type is IgG. If the type of heavy chain constant region of the antibody is delta, the antibody type is IgD. If the type of heavy chain constant region of the antibody is epsilon, the antibody type is IgE. If the type of heavy chain constant region of the antibody is mu, the antibody type is IgM.

[0155] Among the above five types of antibodies, IgG and IgA are further divided into subclasses. If the antibody is a human antibody, if the type of heavy chain constant region of the antibody is gamma 1 (γ1), the type of antibody is IgG1; if it is gamma 2 (γ2), the type of antibody is IgG2; if it is gamma 3 (γ3), the type of antibody is IgG3; and if it is gamma 4 (γ4), the type of antibody is IgG4. If the heavy chain constant region of the human antibody is alpha 1 (α1), the type of antibody is IgA1; and if it is alpha 2 (α2), the type of antibody is IgA2.

[0156] Type 1 of Antibodies - IgG

[0157] Immunoglobulin G (IgG), a type of antibody, is a monomer that includes two heavy chains and two light chains. In one specific example, the two heavy chains included in the IgG may be the same heavy chain. Specifically, the two heavy chains included in the IgG may be composed of the same amino acid sequence. In another specific example, the two heavy chains included in the IgG may be different heavy chains. Specifically, the amino acid sequences of the two heavy chains included in the IgG may be different from each other. Each heavy chain of the IgG includes four immunoglobulin domains (VH, CH1, CH2, and CH3), and does not include CH4.

[0158] In one specific example, the two light chains included in the IgG may be the same light chain. Specifically, the two light chains included in the IgG may be composed of the same amino acid sequence. In another specific example, the two light chains included in the IgG may be different light chains. Specifically, the amino acid sequences of the two light chains included in the IgG may be different from each other.

[0159] For convenience in explaining the structure of IgG, in the table of contents of <<Types of Antibodies 1 - IgG>>, the two heavy chains and two light chains contained in IgG are respectively called the first heavy chain, second heavy chain, first light chain, and second light chain. In the IgG, one heavy chain is connected to one light chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, and the second heavy chain is connected to the second light chain by a disulfide bond. The IgG forms an overall Y-shaped quaternary structure.

[0160] If the subtype of IgG is IgG1, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0161] If the subtype of IgG is IgG2, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0162] When the subtype of IgG is IgG3, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0163] If the subtype of IgG is IgG4, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0164] Antibody Type 2 - IgA

[0165] Immunoglobulin A (IgA), a type of antibody, has two subtypes: IgA1 and IgA2.

[0166] IgA1 is a monomer comprising two heavy chains and two light chains. In one specific example, the two heavy chains included in the Ig A1 may be the same heavy chain. Specifically, the two heavy chains included in the Ig A1 may be composed of the same amino acid sequence. In another specific example, the two heavy chains included in the Ig A1 may be different heavy chains. Specifically, the amino acid sequences of the two heavy chains included in the Ig A1 may be different from each other. Each heavy chain of the Ig A1 includes four immunoglobulin domains (VH, CH1, CH2, and CH3) and does not include CH4.

[0167] In one specific example, the two light chains included in the Ig A1 may be the same light chain. Specifically, the two light chains included in the Ig A1 may be composed of the same amino acid sequence. In another specific example, the two light chains included in the Ig A1 may be different light chains. Specifically, the amino acid sequences of the two light chains included in the Ig A1 may be different from each other.

[0168] For convenience in explaining the structure of IgA1, in the table of contents of <<Types of Antibodies 2 - IgA>>, the two heavy chains and two light chains included in IgA1 are respectively called the first heavy chain, the second heavy chain, the first light chain, and the second light chain. In the Ig A1, one heavy chain is connected to one light chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, and the second heavy chain is connected to the second light chain by a disulfide bond. The first heavy chain and the second heavy chain are connected by a disulfide bond at the hinge portion of each heavy chain. The Ig A1 forms an overall Y-shaped quaternary structure.

[0169] IgA2 is a dimer and includes four heavy chains and four light chains. In one specific example, the four heavy chains included in the Ig A2 may be the same heavy chain. Specifically, the four heavy chains included in the Ig A2 may be composed of the same amino acid sequence. In another specific example, the four heavy chains included in the Ig A2 may be different heavy chains. Specifically, the amino acid sequences of the four heavy chains included in the Ig A2 may be different from each other. Each heavy chain of the Ig A2 includes four immunoglobulin domains (VH, CH1, CH2, and CH3) and does not include CH4.

[0170] In one specific example, the four light chains included in the Ig A2 may be the same light chain. Specifically, the four light chains included in the Ig A2 may be composed of the same amino acid sequence. In another specific example, the four light chains included in the Ig A2 may be different light chains. Specifically, the amino acid sequences of the four light chains included in the Ig A2 may be different from each other.

[0171] For convenience in explaining the structure of IgA2, in the explanation below in the table of contents of <<Types of Antibodies 2 - IgA>>, the four heavy chains and four light chains included in IgA2 are respectively called the first heavy chain, second heavy chain, third heavy chain, fourth heavy chain, first light chain, second light chain, third light chain, and fourth light chain. In the above Ig A2, one light chain is connected to one heavy chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, the second heavy chain is connected to the second light chain by a disulfide bond, the third heavy chain is connected to the third light chain by a disulfide bond, and the fourth heavy chain is connected to the fourth light chain by a disulfide bond. The first and second heavy chains are connected by a disulfide bond at the hinge portion of each heavy chain. The third and fourth heavy chains are connected by a disulfide bond at the hinge portion of each heavy chain. The portion comprising the first heavy chain, second heavy chain, first light chain, and second light chain forms an overall Y-shaped quaternary structure. Similarly, the portion comprising the third heavy chain, fourth heavy chain, third light chain, and fourth light chain forms an overall Y-shaped quaternary structure.

[0172] The C-terminal portions of the first and second heavy chains are connected to the C-terminal portions of the third and fourth heavy chains by the J chain (joining chain, J chain).

[0173] If the subtype of IgA is IgA1, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0174] If the subtype of IgA is IgA2, the sequences of CH1, CH2, CH3, and the hinge region may each be a publicly known amino acid sequence known in the art.

[0175] Type 3 of Antibodies - IgE

[0176] Immunoglobulin E (IgE), a type of antibody, is a monomer that includes two heavy chains and two light chains. In one specific example, the two heavy chains included in the IgE may be the same heavy chain. Specifically, the two heavy chains included in the IgE may be composed of the same amino acid sequence. In another specific example, the two heavy chains included in the IgE may be different heavy chains. Specifically, the amino acid sequences of the two heavy chains included in the IgE may be different from each other. Each heavy chain of the IgE includes five immunoglobulin domains (VH, CH1, CH2, CH3, and CH4).

[0177] In one specific example, the two light chains included in the IgE may be the same light chain. Specifically, the two light chains included in the IgE may be composed of the same amino acid sequence. In another specific example, the two light chains included in the IgE may be different light chains. Specifically, the amino acid sequences of the two light chains included in the IgE may be different from each other.

[0178] For convenience in explaining the structure of IgE, in the table of contents of <<Types of Antibodies 3 - IgE>>, the two heavy chains and two light chains included in IgE are respectively called the first heavy chain, the second heavy chain, the first light chain, and the second light chain. In the IgE, one heavy chain is connected to one light chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, and the second heavy chain is connected to the second light chain by a disulfide bond. The first heavy chain and the second heavy chain are connected by a disulfide bond at the hinge portion of each heavy chain. The IgE forms an overall Y-shaped quaternary structure.

[0179] The sequences of CH1, CH2, CH3, CH4 and hinge portions of IgE may each be amino acid sequences known in the art.

[0180] Type 4 of Antibodies - IgM

[0181] Immunoglobulin M (IgM), a type of antibody, is a pentamer and includes 10 heavy chains and 10 light chains. In one specific example, the 10 heavy chains included in the IgM may be the same heavy chain. Specifically, the 10 heavy chains included in the IgM may be composed of the same amino acid sequence. In another specific example, the 10 heavy chains included in the IgM may be different heavy chains. Specifically, the amino acid sequences of the 10 heavy chains included in the IgM may be different from each other. Each heavy chain of the IgM includes 5 immunoglobulin domains (VH, CH1, CH2, CH3, and CH4).

[0182] In one specific example, the ten light chains included in the IgM may be identical light chains. Specifically, the ten light chains included in the IgM may be composed of the same amino acid sequence. In another specific example, the ten light chains included in the IgM may be different light chains. Specifically, the amino acid sequences of the ten light chains included in the IgM may be different from each other.

[0183] For convenience in explaining the structure of IgM, in the table of contents of <<Types of Antibodies 4 - IgM>>, the 10 heavy chains and 10 light chains included in IgM are respectively called the 1st heavy chain, 2nd heavy chain, 3rd heavy chain, 4th heavy chain, 5th heavy chain, 6th heavy chain, 7th heavy chain, 8th heavy chain, 9th heavy chain, 10th heavy chain, 1st light chain, 2nd light chain, 3rd light chain, 4th light chain, 5th light chain, 6th light chain, 7th light chain, 8th light chain, 9th light chain, and 10th light chain. In the above IgM, one heavy chain is connected to one light chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, the second heavy chain is connected to the second light chain by a disulfide bond, the third heavy chain is connected to the third light chain by a disulfide bond, the fourth heavy chain is connected to the fourth light chain by a disulfide bond, the fifth heavy chain is connected to the fifth light chain by a disulfide bond, the sixth heavy chain is connected to the sixth light chain by a disulfide bond, the seventh heavy chain is connected to the seventh light chain by a disulfide bond, the eighth heavy chain is connected to the eighth light chain by a disulfide bond, the ninth heavy chain is connected to the ninth light chain by a disulfide bond, and the tenth heavy chain is connected to the tenth light chain by a disulfide bond.

[0184] The first and second heavy chains are linked by disulfide bonds at the hinge regions of each heavy chain. The third and fourth heavy chains are linked by disulfide bonds at the hinge regions of each heavy chain. The fifth and sixth heavy chains are linked by disulfide bonds at the hinge regions of each heavy chain. The seventh and eighth heavy chains are linked by disulfide bonds at the hinge regions of each heavy chain. The ninth and tenth heavy chains are linked by disulfide bonds at the hinge regions of each heavy chain. The first and second heavy chains, the third and fourth heavy chains, the fifth and sixth heavy chains, the seventh and eighth heavy chains, and the ninth and tenth heavy chains each form an overall Y-shaped quaternary structure.

[0185] The C-terminal portions of the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, and tenth heavy chains are connected by the J chain (joining chain, J chain).

[0186] The sequences of CH1, CH2, CH3, CH4 and hinge portion of IgM may each be a publicly known amino acid sequence known in the art.

[0187] Antibody Type 5 - IgD

[0188] Immunoglobulin D (IgD), a type of antibody, is a monomer that includes two heavy chains and two light chains. In one specific example, the two heavy chains included in the IgD may be the same heavy chain. Specifically, the two heavy chains included in the IgD may be composed of the same amino acid sequence. In another specific example, the two heavy chains included in the IgD may be different heavy chains. Specifically, the amino acid sequences of the two heavy chains included in the IgD may be different from each other. Each heavy chain of the IgD includes four immunoglobulin domains (VH, CH1, CH2, and CH3), and does not include CH4.

[0189] In one specific example, the two light chains included in the IgD may be the same light chain. Specifically, the two light chains included in the IgD may be composed of the same amino acid sequence. In another specific example, the two light chains included in the IgD may be different light chains. Specifically, the amino acid sequences of the two light chains included in the IgD may be different from each other.

[0190] For convenience in explaining the structure of IgD, in the table of contents of <<Types of Antibodies 5 - IgD>>, the two heavy chains and two light chains included in IgD are respectively called the first heavy chain, the second heavy chain, the first light chain, and the second light chain. In the IgD, one heavy chain is connected to one light chain by a disulfide bond. That is, the first heavy chain is connected to the first light chain by a disulfide bond, and the second heavy chain is connected to the second light chain by a disulfide bond. The first heavy chain and the second heavy chain are connected by a disulfide bond at the hinge portion of each heavy chain. The IgD forms an overall Y-shaped quaternary structure.

[0191] The sequences of CH1, CH2, CH3, and hinge portions of IgD may each be amino acid sequences known in the art.

[0192] fibrosis

[0193] As used herein, "fibrosis" refers collectively to fibrosis-associated diseases, fibrosis-associated conditions, and fibrosis-associated symptoms, unless otherwise specified. For example, the fibrosis-associated diseases, fibrosis-associated conditions, and fibrosis-associated symptoms include hypertrophic scar; systemic sclerosis; multiple cancers; pulmonary arterial hypertension; glial scar; Alzheimer's; cardiac fibrosis; hypertrophic cardiomyopathy; cardiac dysfunction; valvular disease; arrhythmia; myelofibrosis; myelodysplastic syndrome; chronic myelogenous leukemia; cirrhosis; portal hypertension; hepatocellular carcinoma; Nonalcoholic steatohepatitis (NASH); intestinal fibrosis; enteropathies; inflammatory bowel disease; arthrofibrosis; subretinal fibrosis; epithelial fibrosis; vision loss; idiopathic pulmonary fibrosis; cystic fibrosis; pulmonary hypertension; thromboembolic disease; emphysema;Renal fibrosis; cystic fibrosis; nephrogenic systemic fibrosis; chronic kidney disease; renal anemia; retroperitoneal fibrosis; mediastinal fibrosis; pancreatic fibrosis; cystic fibrosis; chronic pancreatitis; biliary duct obstruction; autoimmune-interstitial lung disease, including ankylosing spondylitis and rheumatoid arthritis; connective tissue disease-interstitial lung disease, including rheumatoid arthritis-interstitial lung disease; and non-idiopathic pulmonary fibrosis-interstitial lung disease, but is not limited thereto. In addition, the fibrosis may be fibrosis occurring in tissues such as skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and / or pancreas, but is not limited thereto. The above terms include all meanings that can be recognized by a person skilled in the art and can be appropriately interpreted according to the context.;

[0194]

[0195] Background technology

[0196] Fibrosis occurs in various tissues and has been reported as a pathological consequence of various chronic inflammatory diseases. While treatments have been developed for fibrosis in some tissues, it is difficult to predict whether treatments for fibrosis in other tissues will also be effective in dermal fibrosis. For example, nintedanib and pirfenidone are marketed as treatments for idiopathic pulmonary fibrosis, a type of fibrosis. However, neither nintedanib nor pirfenidone is currently approved for the treatment of fibrosis in tissues other than the lungs.

[0197] The inventors of the present disclosure have discovered the use of CSF3 inhibitors for treating pulmonary fibrosis. Details regarding the use of CSF3 inhibitors for treating pulmonary fibrosis discovered by the inventors of the present disclosure are disclosed in the literature (WO 2020 / 159243). However, as previously described, it is difficult to predict that a fibrosis treatment targeting lung tissue will also have a therapeutic effect on fibrosis in other tissues. Therefore, it is difficult to predict that CSF3 inhibitors can have a therapeutic effect on fibrosis regardless of tissue based solely on the information disclosed in the published literature (WO 2020 / 159243). For example, the published literature (WO 2020 / 159243) only describes that CSF3 inhibitors exhibit a therapeutic effect on pulmonary fibrosis, and does not describe whether CSF3 inhibitors can exhibit a therapeutic effect on fibrosis in other tissues.

[0198] Therefore, there is an unmet need for therapeutics that can treat fibrosis regardless of tissue type, and related research is required.

[0199]

[0200] 1. Mechanism of action of CSF3 inhibitors that restore fibrotic tissue to normal tissue

[0201] 1.1. Background - The effect of CSF3 inhibitors on returning fibrotic tissue to normal tissue.

[0202] As described in this background art, the inventors of the present disclosure have discovered the use of CSF3 inhibitors in the treatment of pulmonary fibrosis. Specifically, the inventors of the present disclosure discovered that when the function of CSF3 in fibrotic lung tissue is inhibited using an antibody, the fibrotic lung tissue is restored to normal lung tissue and fibrosis is cured (WO 2020 / 159243). This therapeutic effect of reverting fibrotic lung tissue to normal lung tissue has not been confirmed in treatment using commercially available pulmonary fibrosis treatments. Therefore, the inventors of the present disclosure studied and elucidated the mechanism of action of CSF3 inhibitors in reverting fibrotic tissue to normal tissue to specifically understand the cause of the effect of reverting fibrotic lung tissue to normal lung tissue and curing fibrosis. The results are disclosed in the present disclosure.

[0203] 1.2. Overview of the mechanism of action of CSF3 inhibitors

[0204] In one aspect of the present disclosure, a method for restoring fibrotic tissue (or cells) to normal tissue (or cells) is disclosed. In another aspect of the present disclosure, a method for inhibiting extracellular matrix accumulation and epithelial-to-mesenchymal transition (EMT) is disclosed. In this case, the method for restoring fibrotic tissue to normal tissue may include a method for inhibiting extracellular matrix accumulation and EMT. In one specific example, the method for inhibiting extracellular matrix accumulation and EMT may be a method for inhibiting extracellular matrix accumulation and EMT in a target tissue. In this case, the target tissue may be a tissue at risk of developing extracellular matrix accumulation and / or EMT or a tissue in which extracellular matrix accumulation and / or EMT has developed.

[0205] The method for restoring the above fibrotic tissue to normal tissue comprises administering a CSF3 inhibitor to the subject or the subject's tissue. The method for inhibiting the above extracellular matrix accumulation and EMT comprises administering a CSF3 inhibitor to the subject or the subject's tissue.

[0206] In one specific example, the CSF3 inhibitor can inhibit the TGF-β mediated signaling pathway. The TGF-β mediated signaling pathway is a signaling pathway that can induce fibrosis through the interaction between TGF-β and a TGF-β receptor.

[0207] In one specific example, the CSF3 inhibitor can inhibit a non-TGF-β mediated signaling pathway. The non-TGF-β mediated signaling pathway is a signaling pathway that induces fibrosis through the interaction between CSF3 and CSF3R, and is a signaling pathway that induces fibrosis in a separate pathway from the TGF-β mediated signaling pathway. Therefore, the non-TGF-β mediated signaling pathway can also be expressed as a CSF3 associated and non-TGF-β mediated signaling pathway.

[0208] In one specific example, the CSF3 inhibitor can inhibit the positive feedback of CSF3. In one specific example, the CSF3 inhibitor can inhibit the positive feedback of TGF-β. In one specific example, the CSF3 inhibitor can restore fibrotic tissue (or cells) to normal tissue (or cells). In one specific example, the CSF3 inhibitor can inhibit the extracellular matrix accumulation and EMT.

[0209] In one specific example, the subject may be a human or a non-human animal.

[0210] In one specific example, the tissue may be skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and / or pancreas.

[0211] In one specific example, the cell may be a cell of the skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and / or pancreas.

[0212] 1.3. Meaning of CSF3 inhibitors

[0213] Colony-Stimulating Factor 3 (CSF3) of the present disclosure is also referred to as Granulocyte Colony-Stimulating Factor (G-CSF). CSF3 is known to stimulate proliferation, differentiation, aggregation, and terminal cell functional activation of granulocytes in animals, including human subjects. The term "CSF3 inhibitor" of the present disclosure refers to a substance that ultimately inhibits the function of CSF3. The CSF3 inhibitor is not otherwise limited in type, form, or composition, as long as it can inhibit the function of CSF3. Specifically, inhibiting the function of CSF3 means 1) binding to the CSF3 protein to inhibit the function of the CSF3 protein itself, 2) inhibiting the interaction of the CSF3 protein with other molecules, 3) inhibiting the expression of CSF3, thereby blocking the opportunity for CSF3 to function, or 4) inhibiting the function of CSF3 through other direct or indirect pathways.

[0214] In one specific example, the CSF3 inhibitor may be an anti-CSF3 antibody. In another specific example, the CSF3 inhibitor may be a nucleic acid molecule capable of silencing the mRNA of the CSF3 gene or causing RNA interference. Specifically, the nucleic acid molecule may include an antisense oligonucleotide; an interfering RNA (iRNA), such as miRNA, siRNA (small interfering RNA), or shRNA (short hairpin RNA). In another specific example, the CSF3 inhibitor may be a compound that inhibits the function of CSF3.

[0215] 1.4. Mechanism of action of CSF3 inhibitors 1: Inhibition of TGF-β-mediated signaling pathway

[0216] In one specific example, the CSF3 inhibitor can inhibit the TGF-β mediated signaling pathway. In another specific example, the CSF3 inhibitor can inhibit the TGF-β mediated signaling pathway in a tissue (or cell) of the subject. In another specific example, when CSF3 is inhibited, the TGF-β mediated signaling pathway can be inhibited in the tissue (or cell) of the subject. That is, the action or activation of the TGF-β mediated signaling pathway involved in fibrosis can be reduced.

[0217] 1.4.1. Significance of the TGF-β-mediated signaling pathway

[0218] The term “TGF-β mediated signaling pathway” of the present disclosure refers to a signaling pathway by which transforming growth factor beta (TGF-β) induces fibrosis, epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition. Specifically, the TGF-β mediated signaling pathway refers to a signaling pathway by which fibrosis, EMT, FMT, and / or extracellular matrix deposition are induced by the interaction of TGF-β and its receptor (TGF-βR).

[0219] In one specific example, the TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0220] In one specific example, activation of the TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0221] In one specific example, the TGF-β-mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced as the expression level of TGF-β increases.

[0222] In one specific example, the TGF-β-mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced by the interaction (or binding) of TGF-β and its receptor.

[0223] In one specific example, the TGF-β mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced as the level of interaction between TGF-β and its receptor increases.

[0224] In one specific example, the TGF-β may be TGF-β1 (Transforming growth factor beta-1), TGF-β2 (Transforming growth factor beta-2), or TGF-β3 (Transforming growth factor beta-3). In one specific example, the receptor of the TGF-β may be TGF-βRII.

[0225] In one specific example, the TGF-β mediated signaling pathway may be represented as a TGF-β1 mediated signaling pathway, a TGF-β2 mediated signaling pathway, or a TGF-β3 mediated signaling pathway.

[0226] 1.4.2. Relationship between TGF-β-mediated signaling pathway and fibrosis

[0227] The above TGF-β-mediated signaling pathway can cause fibrosis or fibrosis-related diseases, conditions, and / or symptoms (hereinafter referred to as “fibrosis”). Specifically, activation of the above TGF-β-mediated signaling pathway can cause fibrosis.

[0228] At this time, activation of the TGF-β-mediated signaling pathway can induce fibrosis through any of the following actions:

[0229] For example, activation of the TGF-β-mediated signaling pathway can induce TGF-β-related immune and / or inflammatory responses.

[0230] In one specific example, activation of the TGF-β mediated signaling pathway can induce activation of fibroblasts and myofibroblasts.

[0231] In one specific example, activation of the TGF-β mediated signaling pathway can promote extracellular matrix production and / or secretion by myofibroblasts.

[0232] In one specific example, activated myofibroblasts, due to activation of the TGF-β-mediated signaling pathway, can actively secrete extracellular matrix, particularly fibrous connective tissue such as collagen.

[0233] In one specific example, activation of the TGF-β-mediated signaling pathway can maintain the activity of myofibroblasts, thereby allowing extracellular matrix such as collagen to continuously accumulate within the tissue.

[0234] In one specific example, activation of the TGF-β-mediated signaling pathway can induce epithelial to mesenchymal transition (EMT) of tissue epithelial cells.

[0235] Activation of these TGF-β-mediated signaling pathways can lead to excessive formation of fibrous connective tissue in tissues, which can induce fibrosis.

[0236] 1.4.3. Biomarkers Associated with the TGF-β-Mediated Signaling Pathway

[0237] Therefore, activation of the TGF-β-mediated signaling pathway affects the expression levels of factors associated with fibrosis. For example, activation of the TGF-β-mediated signaling pathway can increase the expression levels of factors that influence the induction / exacerbation of fibrosis in the tissue where fibrosis has occurred or in the vicinity of the tissue where fibrosis has occurred. For example, activation of the TGF-β-mediated signaling pathway can decrease the expression levels of factors that influence the alleviation / inhibition of fibrosis.

[0238] In one specific example, activation of the TGF-β mediated signaling pathway may increase the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, and IL-11.

[0239] In one specific example, activation of the TGF-β-mediated signaling pathway can induce expression of the extracellular matrix. Specifically, activation of the TGF-β-mediated signaling pathway can increase the expression level of any one or more of collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN).

[0240] In one specific example, activation of the TGF-β-mediated signaling pathway can increase the expression level of hyaluronan synthase (HAS), wherein the HAS can be HAS1, HAS2, and / or HAS3.

[0241] In one specific example, activation of the TGF-β-mediated signaling pathway can inhibit the expression of enzymes that promote extracellular matrix degradation. Specifically, activation of the TGF-β-mediated signaling pathway can reduce the expression level of matrix metalloproteinase (MMP). In this case, the MMP may be one or more selected from MMP2, MMP9, and MMP13.

[0242] In one specific example, activation of the TGF-β mediated signaling pathway can increase the expression level of tissue inhibitor of metalloproteinase (TIMP).

[0243] In one specific example, activation of the TGF-β mediated signaling pathway can reduce the expression level of E-cadherin.

[0244] In one specific example, activation of the TGF-β mediated signaling pathway may increase the expression level of any one or more of Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0245] In one embodiment, inhibition of the TGF-β mediated signaling pathway may result in inhibition of the expression or a decrease in the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0246] In another embodiment, inhibition of the TGF-β mediated signaling pathway may induce or increase the expression level of one or more of matrix metalloproteinase (MMP) and E-cadherin.

[0247] 1.4.4. Mechanism of CSF3 inhibitors inhibiting the TGF-β-mediated signaling pathway

[0248] The CSF3 inhibitor of the present disclosure inhibits the TGF-β-mediated signaling pathway. That is, the CSF3 inhibitor can reduce the action or activation of the TGF-β-mediated signaling pathway involved in fibrosis.

[0249] For example, the CSF3 inhibitor can inhibit the TGF-β mediated signaling pathway by inhibiting the interaction (or binding) of CSF3 and CSF3R.

[0250] In one specific example, the CSF3 inhibitor can inhibit the interaction (or binding) between TGF-β and a TGF-β receptor. In one specific example, the CSF3 inhibitor can inhibit the interaction between TGF-β and a TGF-β receptor by inhibiting the interaction between CSF3 and CSF3R. Therefore, the CSF3 inhibitor of the present disclosure can inhibit the interaction (or binding) between TGF-β and a TGF-β receptor.

[0251] As another example, the TGF-β-mediated signaling pathway may also be inhibited by reducing the activity of the non-TGF-β-mediated signaling pathway by the CSF3 inhibitor.

[0252] In one specific example, the CSF3 inhibitor may also inhibit non-TGF-β mediated signaling pathways, and inhibition of the non-TGF-β mediated signaling pathway may inhibit the interaction of TGF-β with TGF-β receptors.

[0253] As another example, the CSF3 inhibitor can inhibit epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition by inhibiting the TGF-β mediated signaling pathway. In another specific embodiment, the CSF3 inhibitor can inhibit EMT, FMT, and / or extracellular matrix deposition by inhibiting the interaction of CSF3 and CSF3R. In another specific embodiment, the CSF3 inhibitor also inhibits non-TGF-β mediated signaling pathways, and EMT, FMT, and / or extracellular matrix deposition can be inhibited by inhibition of the non-TGF-β mediated signaling pathways.

[0254] In one embodiment, the CSF3 inhibitor can inhibit the expression or reduce the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin, the expression levels of which are increased due to activation of the TGF-β mediated signaling pathway.

[0255] In another embodiment, the CSF3 inhibitor can induce or increase the expression level of one or more of matrix metalloproteinases (MMPs) and E-cadherin, the expression levels of which are reduced due to activation of the TGF-β-mediated signaling pathway.

[0256] 1.5. Mechanism of action of CSF3 inhibitors 2: Inhibition of non-TGF-β-mediated signaling pathways

[0257] The CSF3 inhibitor of the present disclosure can inhibit a non-TGF-β mediated signaling pathway.

[0258] In one specific example, the CSF3 inhibitor can inhibit the non-TGF-β mediated signaling pathway in the tissue (or cell) of the subject. In another specific example, inhibition of CSF3 can inhibit the non-TGF-β mediated signaling pathway in the tissue (or cell) of the subject. That is, the CSF3 inhibitor can reduce the action or activation of the non-TGF-β mediated signaling pathway involved in fibrosis.

[0259] 1.5.1. Implications of non-TGF-β-mediated signaling pathways

[0260] The term “non-TGF-β mediated signaling pathway” of the present disclosure refers to a signaling pathway by which fibrosis, epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition are induced by CSF3, which is a separate pathway from the TGF-β mediated signaling pathway activated by the interaction of TGF-β and a TGF-β receptor. Specifically, the non-TGF-β mediated signaling pathway refers to a signaling pathway by which fibrosis, EMT, FMT, and / or extracellular matrix deposition are induced by the interaction of CSF3 and CSF3R. Therefore, in the present specification, the “non-TGF-β-mediated signaling pathway” may also be expressed as “CSF3-associated and non-TGF-β-mediated signaling pathways.”

[0261] In one specific example, the CSF3-associated and non-TGF-β mediated signaling pathways may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation within tissues.

[0262] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0263] In one specific example, the CSF3-associated and non-TGF-β-mediated signaling pathway may be a signaling pathway that induces fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation as the expression level of CSF3 increases.

[0264] In one specific example, the CSF3-associated and non-TGF-β mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced by the interaction (or binding) of CSF3 and CSF3R.

[0265] In one specific example, the CSF3-associated and non-TGF-β mediated signaling pathway may be a signaling pathway that induces fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation as the level of interaction between CSF3 and CSF3R increases.

[0266] 1.5.2. Relationship between non-TGF-β-mediated signaling pathways and fibrosis

[0267] The above CSF3-associated and non-TGF-β-mediated signaling pathways can induce fibrosis. Specifically, activation of the above CSF3-associated and non-TGF-β-mediated signaling pathways can induce fibrosis.

[0268] At this time, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can induce fibrosis through any of the following actions:

[0269] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways may induce immune and / or inflammatory responses that do not involve TGF-β.

[0270] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways can induce activation of fibroblasts and myofibroblasts.

[0271] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways may promote extracellular matrix production and / or secretion by myofibroblasts.

[0272] In one specific example, activated myofibroblasts, due to activation of the CSF3-associated and non-TGF-β-mediated signaling pathways, can actively secrete extracellular matrix, particularly fibrous connective tissue such as collagen.

[0273] In one specific example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways may maintain the activity of myofibroblasts, thereby allowing the continued deposition of extracellular matrix, such as collagen, within the tissue.

[0274] In one specific example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can induce epithelial to mesenchymal transition (EMT) of tissue epithelial cells.

[0275] Likewise, activation of CSF3-associated and non-TGF-β-mediated signaling pathways can also lead to excessive formation of fibrous connective tissue in tissues, inducing fibrosis.

[0276] 1.5.3. Biomarkers Associated with Non-TGF-β-Mediated Signaling Pathways

[0277] Therefore, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways affects the expression levels of factors associated with fibrosis. For example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can increase the expression levels of factors that influence the induction / exacerbation of fibrosis. For example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can decrease the expression levels of factors that influence the alleviation / inhibition of fibrosis.

[0278] In one specific example, the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11 may be increased by the CSF3-associated and non-TGF-β-mediated signaling pathway. In one specific example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathway may increase the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11.

[0279] In one specific example, the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11 may be increased by CSF3. In this case, the increase in the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, and IL-11 by CSF3 may occur without the involvement of a TGF-β-mediated signaling pathway activated by the interaction of TGF-β and a TGF-β receptor.

[0280] In one specific example, the expression of the extracellular matrix can be induced. Specifically, the expression level of one or more of collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN) can be increased.

[0281] In one specific example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can increase the expression level of hyaluronan synthase (HAS), wherein the HAS can be HAS1, HAS2, and / or HAS3.

[0282] In one specific example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can inhibit the expression of enzymes that promote extracellular matrix degradation. Specifically, activation of the non-TGF-β-mediated signaling pathway can reduce the expression level of matrix metalloproteinases (MMPs). In this case, the MMPs may be at least one selected from MMP2, MMP9, and MMP13.

[0283] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways can increase the expression level of tissue inhibitor of metalloproteinase (TIMP).

[0284] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways can reduce the expression level of E-cadherin.

[0285] In one specific example, activation of the CSF3-associated and non-TGF-β mediated signaling pathways can increase the expression levels of any one or more of Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0286] In one embodiment, inhibition of the CSF3-associated and non-TGF-β mediated signaling pathways may result in inhibition of expression or a decrease in the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0287] In one embodiment, inhibition of the CSF3-associated and non-TGF-β mediated signaling pathways may induce or increase the expression level of one or more of matrix metalloproteinases (MMPs) and E-cadherin.

[0288] 1.5.4. Mechanism of CSF3 inhibitors to inhibit non-TGF-β-mediated signaling pathways

[0289] The CSF3 inhibitor of the present disclosure inhibits the CSF3-mediated and non-TGF-β-mediated signaling pathways. That is, the CSF3 inhibitor can reduce the operation or activation of the CSF3-mediated but not TGF-β-mediated signaling pathway involved in fibrosis.

[0290] In one specific example, in the CSF3-associated and non-TGF-β-mediated signaling pathway process, the TGF-β-mediated signaling pathway can be triggered (or activated) by signaling accordingly. That is, a CSF3 inhibitor can inhibit the CSF3-associated and non-TGF-β-mediated signaling pathway, thereby inhibiting the TGF-β-mediated signaling pathway.

[0291] In one specific example, the CSF3 inhibitor can inhibit the CSF3-associated and non-TGF-β mediated signaling pathway by inhibiting the interaction (or binding) of CSF3 and CSF3R.

[0292] In one specific example, the CSF3 inhibitor inhibits CSF3-related and non-TGF-β mediated signaling pathways by inhibiting CSF3 expression or function, thereby inhibiting epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition. In another specific example, the CSF3 inhibitor inhibits EMT, FMT, and / or extracellular matrix deposition by inhibiting the interaction of CSF3 and CSF3R.

[0293] In one specific example, the CSF3 inhibitor can inhibit the expression or reduce the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin, the expression levels of which are increased due to activation of the CSF3-associated and non-TGF-β-mediated signaling pathways.

[0294] In one specific example, the CSF3 inhibitor can induce or increase the expression level of one or more of matrix metalloproteinases (MMPs) and E-cadherin, the expression levels of which are reduced due to activation of the CSF3-associated and non-TGF-β mediated signaling pathways.

[0295] 1.6. Mechanism of action of CSF3 inhibitors 3: Inhibition of positive feedback loops

[0296] In one specific example, the CSF3 inhibitor may inhibit one or more of the following positive-feedback loops:

[0297] Positive feedback loop of CSF3,

[0298] The positive feedback loop of TGF-β, and

[0299] Positive-feedback loop between CSF3 and TGF-β.

[0300] In another specific example, the CSF3 inhibitor can inhibit the positive feedback loop of CSF3, the positive feedback loop of TGF-β, and / or the reciprocal positive feedback loop between CSF3 and TGF-β in the tissue (or cell) of the subject, i.e., reduce the action of the positive feedback loops.

[0301] 1.6.1. The significance of the positive feedback loop of CSF3

[0302] The term "positive feedback loop of CSF3" in the present disclosure refers to a signaling pathway in which an increase in CSF3 further induces CSF3 expression. In other words, the positive feedback loop of CSF3 refers to a process in which CSF3 causes more CSF3 to be produced, and the produced CSF3 in turn causes more CSF3 to be produced. In one specific example, activation of the positive feedback loop of CSF3 can further induce CSF3 expression or increase the expression level of CSF3. Therefore, the "positive feedback loop of CSF3" in the present specification may also be referred to as a "self-positive feedback loop of CSF3."

[0303] In one specific example, the positive feedback loop of CSF3 may be activated through one or more of the following processes:

[0304] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by CSF3;

[0305] Increased expression of CSF3 and / or TGF-β by activated TGF-β-mediated signaling pathway and / or activated non-TGF-β-mediated signaling pathway;

[0306] Further activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3;

[0307] TGF-β-mediated signaling pathways are further activated by expressed TGF-β; and

[0308] The expression levels of CSF3 and / or TGF-β are further increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway.

[0309] At this time, the process of “the expression amount of CSF3 and / or TGF-β is increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway is activated by the expressed CSF3; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression amount of CSF3 is increased may be activated due to this repeating process (or loop structure).

[0310] 1.6.2. Implications of the positive feedback loop of TGF-β

[0311] The term “positive feedback loop of TGF-β” in the present disclosure refers to a signaling pathway in which an increase in TGF-β further induces the expression of TGF-β. In other words, the positive feedback loop of TGF-β refers to a process in which more TGF-β is produced due to TGF-β, and the produced TGF-β in turn produces more TGF-β. In one specific example, activation of the positive feedback loop of TGF-β can induce the expression of TGF-β or increase the expression level of TGF-β. Therefore, the “positive feedback loop of TGF-β” in the present specification may also be expressed as a “self-positive feedback loop of TGF-β.”

[0312] In one specific example, the positive-feedback loop of TGF-β can be activated through one or more of the following processes:

[0313] TGF-β-mediated signaling pathway is activated by TGF-β;

[0314] Increased expression of CSF3 and / or TGF-β by activated TGF-β-mediated signaling pathway;

[0315] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3;

[0316] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0317] The expression levels of CSF3 and / or TGF-β are further increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway.

[0318] At this time, the process of “the expression amount of CSF3 and / or TGF-β is increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway is activated by the expressed CSF3; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression amount of TGF-β is increased may be activated due to this repeating process (or loop structure).

[0319] 1.6.3. Implications of the reciprocal positive feedback loop between CSF3 and TGF-β

[0320] The term “mutual positive-feedback loop between CSF3 and TGF-β” in the present disclosure refers to a reciprocal relationship between CSF3 and TGF-β that influences the increase in each other’s expression, and refers to a signaling pathway in which i) the expression of TGF-β is induced by an increase in CSF3, and ii) the expression of CSF3 is induced by an increase in TGF-β. In other words, the mutual positive-feedback loop between CSF3 and TGF-β refers to a process in which more TGF-β is produced due to CSF3, and the produced TGF-β in turn produces more CSF3. Alternatively, the mutual positive-feedback loop between CSF3 and TGF-β refers to a process in which more CSF3 is produced due to TGF-β, and the produced CSF3 in turn produces more TGF-β. In one specific example, activation of the reciprocal positive-feedback loop between CSF3 and TGF-β can induce expression of CSF3 and TGF-β or increase the expression levels of CSF3 and TGF-β.

[0321] In one specific example, the reciprocal positive-feedback loop between CSF3 and TGF-β can be activated through one or more of the following processes:

[0322] Activation of TGF-β-mediated signaling pathways and / or activation of CSF3-associated and non-TGF-β-mediated signaling pathways by CSF3;

[0323] Increased expression of CSF3 and / or TGF-β by activated TGF-β-mediated signaling pathway and / or activated CSF3-associated and non-TGF-β-mediated signaling pathway;

[0324] Activation of TGF-β-mediated signaling pathways and / or CSF3-associated and non-TGF-β-mediated signaling pathways by expressed CSF3;

[0325] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0326] Increased expression of CSF3 and TGF-β by activated TGF-β-mediated signaling pathway and / or activated CSF3-associated and non-TGF-β-mediated signaling pathway.

[0327] At this time, the process of “the expression levels of CSF3 and TGF-β are increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway are activated by the expressed CSF3; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression levels of CSF3 and TGF-β are increased may be activated due to this repeating process (or loop structure).

[0328]

[0329] In another specific example, the reciprocal positive feedback loop between CSF3 and TGF-β can be activated through one or more of the following processes:

[0330] TGF-β-mediated signaling pathway is activated by TGF-β;

[0331] Increased expression of CSF3 and / or TGF-β by activated TGF-β-mediated signaling pathway;

[0332] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3;

[0333] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0334] Increased expression of CSF3 and TGF-β by activated TGF-β-mediated signaling pathway and / or activated non-TGF-β-mediated signaling pathway.

[0335] At this time, the process of “the expression levels of CSF3 and TGF-β are increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway are activated by the expressed CSF3; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression levels of CSF3 and TGF-β are increased may be activated due to this repeating process (or loop structure).

[0336] 1.6.4. Mechanism of inhibition of positive feedback loops by CSF3 inhibitors

[0337] The CSF3 inhibitor of the present disclosure may inhibit one or more of the positive feedback loops described above.

[0338] For example, both the positive feedback loop of CSF3 and the positive feedback loop of TGF-β expression can be inhibited. Similarly, the CSF3 inhibitor can inhibit the reciprocal positive feedback loop between CSF3 and TGF-β.

[0339] In one specific example, the CSF3 inhibitor can inhibit both the expression of CSF3 and the expression of TGF-β. In another specific example, the CSF3 inhibitor can reduce both the expression levels of CSF3 and TGF-β. That is, by inhibiting CSF3, both the expression of CSF3 and the expression of TGF-β can be inhibited.

[0340] In the positive feedback loop of CSF3; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3 and TGF-β, CSF3 and TGF-β are closely related. In this close relationship, (i) when CSF3 is inhibited, the positive feedback loop of CSF3; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3 and TGF-β can be significantly inhibited, whereas (ii) when only TGF-β is inhibited, it is difficult to significantly inhibit the positive feedback loop of CSF3; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3 and TGF-β. That is, compared to when TGF-β is inhibited, when CSF3 is inhibited, the positive feedback loop of CSF3; the positive feedback of TGF-β expression; and the reciprocal positive feedback loop between CSF3 and TGF-β is inhibited to a stronger level.

[0341] For example, when TGF-β is inhibited, the expression levels of CSF3 and TGF-β are reduced, and when CSF3 is inhibited, the expression levels of CSF3 and TGF-β are reduced. In this case, the expression levels of CSF3 and TGF-β are further reduced by CSF3 inhibition compared to TGF-β inhibition.

[0342] This is because inhibition of CSF3 inhibits both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway within the positive-feedback loop of CSF3; the positive-feedback of TGF-β expression; and the mutual positive-feedback loop structure between CSF3 and TGF-β, whereas inhibition of TGF-β inhibits only the TGF-β-mediated signaling pathway.

[0343] 1.7. A Key Regulator in the Signaling Pathway that Leads to Fibrosis: CSF3

[0344] The present inventors have shown for the first time that when CSF3 is suppressed, TGF-β-mediated signaling pathways and non-TGF-β-mediated signaling pathways are suppressed, and when CSF3 is increased, TGF-β-mediated signaling pathways and non-TGF-β-mediated signaling pathways are activated. In other words, it has been revealed for the first time that CSF3 is a key regulator throughout the signaling pathways that induce fibrosis or tissue fibrosis. Based on the above explanation, the present inventors speculate that CSF3 is an upstream regulator of TGF-β based on the experimental data showing that CSF3 is a key regulator in the induction of fibrosis.

[0345] For example, when looking at Experimental Examples 2.3 to 2.7, when comparing i) the CSF3 expression level that is reduced by suppressing only the TGF-β-mediated signaling pathway and ii) the CSF3 expression level that is reduced by suppressing both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway, it can be seen that the reduction in the latter (ii) is greater.

[0346] For example, when looking at Experimental Examples 2.3 to 2.7, when comparing the expression levels of α-SMA, Col1A1, CSF3, IL11, and CSF3R, etc., which are reduced by suppressing only the TGF-β-mediated signaling pathway and ii) the expression levels of α-SMA, Col1A1, CSF3, IL11, and CSF3R, etc., which are reduced by suppressing both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway, it can be seen that the reduction in the latter (ii) is greater.

[0347] For example, looking at Experimental Example 2.5, Fig. 17, and Fig. 18, it can be seen that when the interaction between TGF-β and TGF-β receptor (TGF-βR) was inhibited, the expression levels of fibrosis-related markers were slightly reduced. In contrast, looking at Experimental Example 2.5, Experimental Example 2.6, Fig. 17, Fig. 18, and Fig. 19, it can be seen that when the interaction between CSF3 and CSF3 receptor (CSF3R) was inhibited, the expression levels of fibrosis-related markers were significantly reduced. That is, it can be seen that when the interaction between CSF3 and CSF3 receptor (CSF3R) was inhibited, the expression levels of fibrosis-related markers were closer to the control group (normal group in which fibrosis was not induced) than when the interaction between TGF-β and TGF-β receptor was inhibited.

[0348] 1.8. Effect of CSF3 inhibitors on returning fibrotic tissue to normal tissue

[0349] The CSF3 inhibitor of the present disclosure restores fibrotic tissue (or cells) to normal tissue (or cells). In another specific embodiment, the CSF3 inhibitor causes fibrotic tissue (or cells) to return to normal tissue (or cells).

[0350] Looking at the experimental examples of this specification and FIGS. 15, 20, 26, 28, 29, 41, 42, 43, 45, etc., it can be seen that fibrotic tissue (or cells) reverts to normal tissue (or cells) due to CSF3 inhibition.

[0351] In contrast, commercially available pulmonary fibrosis treatments (e.g., nintedanib or pirfenidone) that inhibit the TGF-β-mediated signaling pathway do not effectively restore fibrotic tissue (or cells) to normal tissue (or cells). In other words, it is widely recognized in the art that these treatments only alleviate fibrotic symptoms and do not restore normal tissue.

[0352] However, the CSF3 inhibitor disclosed herein has a therapeutic effect of restoring fibrotic tissue to normal tissue.

[0353] The reason for the difference in the effectiveness of commercialized pulmonary fibrosis treatments and the CSF3 inhibitors may be that, compared to commercialized pulmonary fibrosis treatments, CSF3 inhibitors not only inhibit the TGF-β-mediated signaling pathway, but also have a greater effect of inhibiting the non-TGF-β-mediated signaling pathway. From another perspective, the reason for the difference in the effectiveness of commercialized pulmonary fibrosis treatments and the CSF3 inhibitors may be that, compared to commercialized pulmonary fibrosis treatments, CSF3 inhibitors have a stronger effect of inhibiting the TGF-β-mediated signaling pathway.

[0354] 1.9. Examples of CSF3 inhibitors

[0355] The CSF3 inhibitor described in the present disclosure is not limited to anything that can inhibit the function of CSF3. The CSF3 inhibitor may 1) interfere with the interaction of the CSF3 protein with other molecules, or 2) inhibit the expression of the CSF3 protein, thereby reducing the absolute amount of the CSF3 protein that interacts with other molecules, thereby interfering with the function of the CSF3 protein. Examples of CSF3 inhibitors capable of such actions include, but are not limited to, a nucleic acid that silences the mRNA of the CSF3 gene, a CRISPR / Cas system that knocks out or knocks down the CSF3 gene, a compound that inhibits the CSF3 protein, or an aptamer that inhibits the CSF3 protein.

[0356] 1.9.1. Nucleic acid that silences the mRNA of the CSF3 gene

[0357] The CSF3 inhibitor may silence the mRNA of the CSF3 gene and thereby interfere with the translation process of the CSF3 protein. In one embodiment, the CSF3 inhibitor may be a nucleic acid that can specifically bind to mRNA transcribed from the CSF3 gene, and may be selected from siRNA (small interfering RNA), shRNA (small hairpin RNA), miRNA (microRNA), and antisense nucleic acid.

[0358] 1.9.2. CRISPR / Cas system targeting the CSF3 gene 1 - nuclease

[0359] The CSF3 inhibitor may knock out the CSF3 gene in the cellular genome, thereby preventing the production of the CSF3 protein. For example, the CRISPR / Cas system, which can act as a target-specific nuclease, can cleave a specific gene in the cellular genome, and can induce an error in the cellular gene repair mechanism (e.g., non-homologous end repair (NHEJ) and / or homologous recombination repair (HDR)) to generate an indel in the specific gene, thereby knocking out the specific gene. In one embodiment, the CSF3 inhibitor may be a composition comprising a CRISPR / Cas system, which comprises: a Cas protein or a nucleic acid encoding the same; and a guide RNA or a nucleic acid encoding the same, wherein the guide RNA can target the CSF3 gene in the cellular genome and interact with the Cas protein to form a complex.

[0360] 1.9.3. CRISPR / Cas system targeting the CSF3 gene 2 - regulator

[0361] The CSF3 inhibitor may suppress the production of CSF3 protein by causing a knockdown effect that suppresses the expression of the CSF3 gene in the cellular genome. For example, in one target-specific embodiment, the CSF3 inhibitor may be a composition comprising a CRISPRi system comprising: a fusion protein in which a Cas protein and at least one gene expression regulatory domain are linked, or a nucleic acid encoding the fusion protein; and a guide RNA and a nucleic acid encoding the same, wherein the guide RNA can target the CSF3 gene in the cellular genome and interact with the fusion protein to form a complex. In this case, the gene expression regulatory domain may be a polypeptide or protein that suppresses the expression of the CSF3 gene in the cellular genome.

[0362] 1.9.4. Substance 1 that inhibits CSF3 protein - Compound

[0363] In one embodiment, the CSF3 inhibitor may be a compound and / or small molecule that functions to prevent the CSF3 protein from interacting with other molecules.

[0364] 1.9.5. Substance 2 that inhibits CSF3 protein - Aptamer

[0365] In one embodiment, the CSF3 inhibitor may be a DNA and / or RNA aptamer that functions to prevent the CSF3 protein from interacting with other molecules.

[0366] 1.9.6. Substance 3 that inhibits CSF3 protein - Antibodies

[0367] In one embodiment, the CSF3 inhibitor may be an anti-CSF3 antibody or a fragment of the anti-CSF3 antibody. The anti-CSF3 antibody refers to an antibody (or immunoglobulin) that recognizes the CSF3 protein as an antigen. The anti-CSF3 antibody refers to an antibody having a function of recognizing all or part of the CSF3 protein; and / or a function of binding to all or part of the CSF3 protein. The fragment of the anti-CSF3 antibody refers to a molecule that includes a part of the anti-CSF3 antibody and has the function of an anti-CSF3 antibody (a function of recognizing and binding to all or part of the CSF3 protein). That is, the anti-CSF3 antibody or the fragment of the anti-CSF3 antibody can recognize and / or bind to all or part of the CSF3 protein. Accordingly, the anti-CSF3 antibody or fragment thereof can bind to the CSF3 protein expressed in the tissue where fibrosis has occurred, thereby preventing the CSF3 protein from interacting with other molecules (e.g., CSF3R), and consequently inhibiting the function of the CSF3 protein. The anti-CSF3 antibody or fragment thereof is not otherwise limited as long as it can inhibit CSF3.

[0368] In one specific example, the anti-CSF3 antibody may be immunoglobulin G (IgG). In this case, the IgG may be selected from IgG1, IgG2, IgG3, and IgG4.

[0369] In one specific example, the anti-CSF3 antibody may be any one of a human antibody, a humanized antibody, and a chimeric antibody.

[0370] In one specific example, the anti-CSF3 antibody may be any one selected from a monoclonal antibody, a polyclonal antibody, a hybridoma monoclonal antibody, and a recombinant monoclonal antibody.

[0371] In one specific example, the anti-CSF3 antibody may be human IgG.

[0372] In one specific example, the anti-CSF3 antibody may be a CSF3 neutralizing antibody.

[0373] In one specific example, the fragment of the anti-CSF3 antibody may be a fragment of a CSF3 neutralizing antibody.

[0374] In one specific example, the fragment of the anti-CSF3 antibody may be selected from F(ab), F(ab'), F(ab')2, monospecific F(ab')2, bispecific F(ab')2, scFv, ScFv-Fc, and sdAb.

[0375] In one specific example, the anti-CSF3 antibody may be one described in the table of contents under <5. Anti-CSF3 Antibody>.

[0376]

[0377] 2. Mechanism of action of CSF3R inhibitors that restore fibrotic tissue to normal tissue

[0378] 2.1. Background - Effect of CSF3R inhibitors on returning fibrotic tissue to normal tissue

[0379] As described in this background art, the inventors of the present disclosure have discovered the use of colony stimulating factor 3 receptor (CSF3R) inhibitors for the treatment of pulmonary fibrosis. Specifically, the inventors of the present disclosure discovered that inhibiting the function of CSF3R in fibrotic lung tissue using an antibody resulted in the restoration of fibrotic lung tissue to normal lung tissue, thereby curing fibrosis (WO 2023 / 038416). This therapeutic effect of restoring fibrotic lung tissue to normal lung tissue is an effect not previously observed in the treatment of pulmonary fibrosis using commercially available therapeutic agents.

[0380] Based on the fact that CSF3 interacts with CSF3R, the inventors of the present invention conducted additional research to confirm whether the effect and mechanism of action when CSF3R is inhibited are the same or similar to those when CSF3 is inhibited as described above.

[0381] That is, the inventors of the present invention further studied the mechanism of action of the CSF3R inhibitor's effect of returning fibrotic tissue to normal tissue in order to determine the cause of the effect of restoring fibrotic lung tissue to normal lung tissue and treating fibrosis, and the results are disclosed herein.

[0382] Through this, it can be seen that even when CSF3R is inhibited, the same mechanism of action is present as when CSF3 is inhibited, for the “TGF-β mediated signaling pathway” and “Non-TGF-β-mediated and CSF3-associated signaling pathways.”

[0383] 2.2. Overview of the mechanism of action of CSF3R inhibitors

[0384] In one aspect of the present disclosure, a method for restoring fibrotic tissue (or cells) to normal tissue (or cells) using a CSF3R inhibitor is disclosed. In another aspect of the present disclosure, a method for inhibiting extracellular matrix accumulation and epithelial to mesenchymal transition (EMT) using a CSF3R inhibitor is disclosed. In this case, the method for restoring fibrotic tissue to normal tissue may include a method for inhibiting extracellular matrix accumulation and EMT. In one specific example, the method for inhibiting extracellular matrix accumulation and EMT may be a method for inhibiting extracellular matrix accumulation and EMT in a target tissue. In this case, the target tissue may be a tissue at risk of developing extracellular matrix accumulation and / or EMT or a tissue in which extracellular matrix accumulation and / or EMT has developed.

[0385] The method for restoring the above fibrotic tissue to normal tissue comprises administering a CSF3R inhibitor to the subject or the subject's tissue. The method for inhibiting the above extracellular matrix accumulation and EMT comprises administering a CSF3R inhibitor to the subject or the subject's tissue.

[0386] In one specific example, the CSF3R inhibitor can inhibit the TGF-β mediated signaling pathway. The TGF-β mediated signaling pathway is a signaling pathway that can induce fibrosis through the interaction between TGF-β and a TGF-β receptor.

[0387] In one specific example, the CSF3R inhibitor can inhibit a non-TGF-β mediated signaling pathway. The non-TGF-β mediated signaling pathway is a signaling pathway that induces fibrosis through the interaction of CSF3 and CSF3R, and is a signaling pathway that induces fibrosis in a separate pathway from the TGF-β mediated signaling pathway. Therefore, the non-TGF-β mediated signaling pathway may be expressed as a CSF3 associated and non-TGF-β mediated signaling pathway. Alternatively, the non-TGF-β mediated signaling pathway may be expressed as a CSF3R associated and non-TGF-β mediated signaling pathway.

[0388] In one specific example, the CSF3R inhibitor can inhibit the positive feedback of the CSF3R. In one specific example, the CSF3R inhibitor can inhibit the positive feedback of the TGF-β. In one specific example, the CSF3R inhibitor can restore the fibrotic tissue (or cell) to a normal tissue (or cell). In one specific example, the CSF3R inhibitor can inhibit the extracellular matrix accumulation and EMT.

[0389] In one specific example, the subject may be a human or a non-human animal.

[0390] In one specific example, the tissue may be skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and / or pancreas.

[0391] In one specific example, the cell may be a cell of the skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and / or pancreas.

[0392] 2.3. Significance of CSF3R inhibitors

[0393] Colony-stimulating factor 3 (CSF3R) of the present disclosure is also referred to as granulocyte colony-stimulating factor (G-CSF). CSF3R is known to stimulate proliferation, differentiation, aggregation, and terminal cell functional activation of granulocytes in animals, including human subjects. The term "CSF3R inhibitor" of the present disclosure refers to a substance that ultimately inhibits the function of CSF3R. The type, form, and composition of the CSF3R inhibitor are not otherwise limited, as long as it can inhibit the function of CSF3R. Specifically, inhibiting the function of CSF3R means 1) binding to the CSF3R protein and inhibiting the function of the CSF3R protein itself, 2) inhibiting the interaction of the CSF3R protein with other molecules, 3) inhibiting the expression of CSF3R and consequently blocking the opportunity for CSF3R to function, or 4) inhibiting the function of CSF3R through other direct or indirect pathways.

[0394] In one specific example, the CSF3R inhibitor may be an anti-CSF3R antibody. In another specific example, the CSF3R inhibitor may be a nucleic acid molecule capable of silencing the mRNA of the CSF3R gene or causing RNA interference. Specifically, the nucleic acid molecule may include an antisense oligonucleotide, an interfering RNA (iRNA), such as miRNA, siRNA (small interfering RNA), or shRNA (short hairpin RNA). In another specific example, the CSF3R inhibitor may be a compound that inhibits the function of CSF3R.

[0395] 2.4. Mechanism of action of CSF3R inhibitors 1: Inhibition of TGF-β-mediated signaling pathway

[0396] In one specific example, the CSF3R inhibitor can inhibit the TGF-β mediated signaling pathway. In another specific example, the CSF3R inhibitor can inhibit the TGF-β mediated signaling pathway in a tissue (or cell) of the subject. In another specific example, when CSF3R is inhibited, the TGF-β mediated signaling pathway can be inhibited in the tissue (or cell) of the subject. That is, the action or activation of the TGF-β mediated signaling pathway involved in fibrosis can be reduced.

[0397] 2.4.1. Significance of the TGF-β-mediated signaling pathway

[0398] The term “TGF-β mediated signaling pathway” of the present disclosure refers to a signaling pathway by which transforming growth factor beta (TGF-β) induces fibrosis, epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition. Specifically, the TGF-β mediated signaling pathway refers to a signaling pathway by which fibrosis, EMT, FMT, and / or extracellular matrix deposition are induced by the interaction of TGF-β and its receptor (TGF-βR).

[0399] In one specific example, the TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0400] In one specific example, activation of the TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0401] In one specific example, the TGF-β-mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced as the expression level of TGF-β increases.

[0402] In one specific example, the TGF-β-mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced by the interaction (or binding) of TGF-β and its receptor.

[0403] In one specific example, the TGF-β mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced as the level of interaction between TGF-β and its receptor increases.

[0404] 2.4.2. Relationship between TGF-β-mediated signaling pathway and fibrosis

[0405] The above TGF-β-mediated signaling pathway can induce fibrosis. Specifically, activation of the above TGF-β-mediated signaling pathway can induce fibrosis.

[0406] At this time, activation of the TGF-β-mediated signaling pathway can induce fibrosis through any of the following actions:

[0407] For example, activation of the TGF-β-mediated signaling pathway can induce TGF-β-related immune and / or inflammatory responses.

[0408] In one specific example, activation of the TGF-β mediated signaling pathway can induce activation of fibroblasts and myofibroblasts.

[0409] In one specific example, activation of the TGF-β mediated signaling pathway can promote extracellular matrix production and / or secretion by myofibroblasts.

[0410] In one specific example, activated myofibroblasts, due to activation of the TGF-β-mediated signaling pathway, can actively secrete extracellular matrix, particularly fibrous connective tissue such as collagen.

[0411] In one specific example, activation of the TGF-β-mediated signaling pathway can maintain the activity of myofibroblasts, thereby allowing extracellular matrix such as collagen to continuously accumulate within the tissue.

[0412] In one specific example, activation of the TGF-β-mediated signaling pathway can induce epithelial to mesenchymal transition (EMT) of tissue epithelial cells.

[0413] Activation of these TGF-β-mediated signaling pathways can lead to excessive formation of fibrous connective tissue in tissues, which can induce fibrosis.

[0414] 2.4.3. Biomarkers Associated with the TGF-β-Mediated Signaling Pathway

[0415] Therefore, activation of the TGF-β-mediated signaling pathway affects the expression levels of factors associated with fibrosis. For example, activation of the TGF-β-mediated signaling pathway can increase the expression levels of factors that influence the induction / exacerbation of fibrosis in the tissue where fibrosis has occurred or in the vicinity of the tissue where fibrosis has occurred. For example, activation of the TGF-β-mediated signaling pathway can decrease the expression levels of factors that influence the alleviation / inhibition of fibrosis.

[0416] In one specific example, activation of the TGF-β mediated signaling pathway may increase the expression level of any one or more of CSF3R, TGF-β, α-SMA, COL1A1, and IL-11.

[0417] In one specific example, activation of the TGF-β-mediated signaling pathway can induce expression of the extracellular matrix. Specifically, activation of the TGF-β-mediated signaling pathway can increase the expression level of any one or more of collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN).

[0418] In one specific example, activation of the TGF-β-mediated signaling pathway can increase the expression level of hyaluronan synthase (HAS), wherein the HAS can be HAS1, HAS2, and / or HAS3.

[0419] In one specific example, activation of the TGF-β-mediated signaling pathway can inhibit the expression of enzymes that promote extracellular matrix degradation. Specifically, activation of the TGF-β-mediated signaling pathway can reduce the expression level of matrix metalloproteinase (MMP). In this case, the MMP may be one or more selected from MMP2, MMP9, and MMP13.

[0420] In one specific example, activation of the TGF-β mediated signaling pathway can increase the expression level of tissue inhibitor of metalloproteinase (TIMP).

[0421] In one specific example, activation of the TGF-β mediated signaling pathway can reduce the expression level of E-cadherin.

[0422] In one specific example, activation of the TGF-β mediated signaling pathway may increase the expression level of any one or more of Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0423] In one embodiment, inhibition of the TGF-β mediated signaling pathway may result in inhibition of expression or a decrease in the expression level of any one or more of CSF3R, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0424] In one embodiment, inhibition of the TGF-β mediated signaling pathway may induce or increase the expression level of one or more of matrix metalloproteinase (MMP) and E-cadherin.

[0425] 2.4.4. Mechanism of CSF3R inhibitors inhibiting the TGF-β-mediated signaling pathway

[0426] The CSF3R inhibitor of the present disclosure inhibits the TGF-β mediated signaling pathway.

[0427] That is, the CSF3R inhibitor can reduce the action or activation of the TGF-β-mediated signaling pathway involved in fibrosis.

[0428] For example, the CSF3R inhibitor can inhibit the TGF-β mediated signaling pathway by inhibiting the interaction (or binding) between CSF3R and CSF3.

[0429] In one specific example, the CSF3R inhibitor can inhibit the interaction (or binding) between TGF-β and a TGF-β receptor. In another specific example, the CSF3R inhibitor can inhibit the interaction between TGF-β and a TGF-β receptor by inhibiting the interaction between CSF3R and CSF3. Accordingly, the CSF3R inhibitor of the present disclosure can inhibit the interaction (or binding) between TGF-β and a TGF-β receptor.

[0430] As another example, the TGF-β-mediated signaling pathway may also be inhibited by reducing the activity of the non-TGF-β-mediated signaling pathway by the CSF3R inhibitor.

[0431] In one specific example, the non-TGF-β mediated signaling pathway may also be inhibited by the CSF3R inhibitor, and the interaction between TGF-β and the TGF-β receptor may be inhibited due to the inhibition of the non-TGF-β mediated signaling pathway.

[0432] As another example, the CSF3R inhibitor can inhibit epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition by inhibiting the TGF-β mediated signaling pathway. In another specific embodiment, the CSF3R inhibitor can inhibit EMT, FMT, and / or extracellular matrix deposition by inhibiting the interaction of CSF3R and CSF3. In another specific embodiment, the non-TGF-β mediated signaling pathway is also inhibited by the CSF3R inhibitor, and EMT, FMT, and / or extracellular matrix deposition can be inhibited by inhibition of the non-TGF-β mediated signaling pathway.

[0433] In one embodiment, the CSF3R inhibitor can inhibit the expression or reduce the expression level of any one or more of CSF3R, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin, the expression levels of which are increased due to activation of the TGF-β mediated signaling pathway.

[0434] In another embodiment, the CSF3R inhibitor can induce or increase the expression level of one or more of matrix metalloproteinases (MMPs) and E-cadherin, the expression levels of which are reduced due to activation of the TGF-β-mediated signaling pathway.

[0435] 2.5. Mechanism of action of CSF3R inhibitors 2: Inhibition of non-TGF-β-mediated signaling pathways

[0436] The CSF3R inhibitor of the present disclosure can inhibit a non-TGF-β mediated signaling pathway.

[0437] In one specific example, the CSF3R inhibitor can inhibit the non-TGF-β mediated signaling pathway in the tissue (or cell) of the subject. In another specific example, inhibition of CSF3R can inhibit the non-TGF-β mediated signaling pathway in the tissue (or cell) of the subject. That is, the CSF3R inhibitor can reduce the action or activation of the non-TGF-β mediated signaling pathway involved in fibrosis.

[0438] 2.5.1. Implications of non-TGF-β-mediated signaling pathways

[0439] The term “non-TGF-β mediated signaling pathway” of the present disclosure refers to a signaling pathway that is separate from the TGF-β mediated signaling pathway activated by the interaction of TGF-β and a TGF-β receptor, and induces fibrosis, epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition by CSF3R. Specifically, the non-TGF-β mediated signaling pathway refers to a signaling pathway that induces fibrosis, EMT, FMT, and / or extracellular matrix deposition by the interaction of CSF3 and CSF3R. Therefore, in the present specification, the “non-TGF-β mediated signaling pathway” may also be expressed as “CSF3-associated and non-TGF-β-mediated signaling pathways.” Alternatively, the non-TGF-β mediated signaling pathway may also be expressed as CSF3R associated and non-TGF-β mediated signaling pathway.

[0440] In one specific example, the non-TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation within the tissue.

[0441] In one specific example, activation of the non-TGF-β mediated signaling pathway may induce fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation.

[0442] In one specific example, the non-TGF-β mediated signaling pathway may be a signaling pathway that induces fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation as the expression level of CSF3R increases.

[0443] In one specific example, the non-TGF-β mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced by the interaction (or binding) of CSF3R and CSF3.

[0444] In one specific example, the non-TGF-β mediated signaling pathway may be a signaling pathway in which fibrosis, epithelial-mesenchymal transition, differentiation of fibroblasts into myofibroblasts, and / or extracellular matrix accumulation are induced as the level of interaction between CSF3R and CSF3 increases.

[0445] 2.5.2. Relationship between non-TGF-β-mediated signaling pathways and fibrosis

[0446] The above non-TGF-β mediated signaling pathway can induce fibrosis. Specifically, activation of the above non-TGF-β mediated signaling pathway can induce fibrosis.

[0447] At this time, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can induce fibrosis through any of the following actions:

[0448] In one specific example, activation of the non-TGF-β mediated signaling pathway may induce an immune and / or inflammatory response that does not involve TGF-β.

[0449] In one specific example, activation of the non-TGF-β mediated signaling pathway can induce activation of fibroblasts and myofibroblasts.

[0450] In one specific example, activation of the non-TGF-β mediated signaling pathway may promote extracellular matrix production and / or secretion by myofibroblasts.

[0451] In one specific example, activated myofibroblasts, due to activation of the non-TGF-β mediated signaling pathway, can actively secrete extracellular matrix, particularly fibrous connective tissue such as collagen.

[0452] In one specific example, activation of the non-TGF-β mediated signaling pathway may maintain the activity of myofibroblasts, thereby allowing the continued deposition of extracellular matrix, such as collagen, within the tissue.

[0453] In one specific example, activation of the non-TGF-β mediated signaling pathway can induce epithelial to mesenchymal transition (EMT) of tissue epithelial cells.

[0454] Likewise, activation of CSF3-associated and non-TGF-β-mediated signaling pathways can also lead to excessive formation of fibrous connective tissue in tissues, thereby inducing fibrosis.

[0455] 2.5.3. Biomarkers Associated with Non-TGF-β-Mediated Signaling Pathways

[0456] Therefore, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways affects the expression levels of factors associated with fibrosis. For example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can increase the expression levels of factors that influence the induction / exacerbation of fibrosis. For example, activation of the CSF3-associated and non-TGF-β-mediated signaling pathways can decrease the expression levels of factors that influence the alleviation / inhibition of fibrosis.

[0457] In one specific example, the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11 may be increased by the non-TGF-β mediated signaling pathway. In one specific example, activation of the non-TGF-β mediated signaling pathway may increase the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11.

[0458] In one specific example, the expression level of any one or more of CSF3, CSF3R, TGF-β, α-SMA, COL1A1, and IL-11 may be increased by CSF3R. In this case, the increase in the expression level of any one or more of CSF3, TGF-β, α-SMA, COL1A1, and IL-11 by CSF3R may occur without the involvement of a TGF-β-mediated signaling pathway activated by the interaction of TGF-β and a TGF-β receptor.

[0459] In one specific example, activation of the non-TGF-β mediated signaling pathway can induce expression of the extracellular matrix. Specifically, activation of the non-TGF-β mediated signaling pathway can increase the expression level of one or more of collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN).

[0460] In one specific example, activation of the non-TGF-β mediated signaling pathway can increase the expression level of hyaluronan synthase (HAS), wherein the HAS can be HAS1, HAS2, and / or HAS3.

[0461] In one specific example, activation of the non-TGF-β mediated signaling pathway can inhibit the expression of enzymes that promote extracellular matrix degradation. Specifically, activation of the non-TGF-β mediated signaling pathway can reduce the expression level of matrix metalloproteinase (MMP). In this case, the MMP may be one or more selected from MMP2, MMP9, and MMP13.

[0462] In one specific example, activation of the non-TGF-β mediated signaling pathway can increase the expression level of tissue inhibitor of metalloproteinase (TIMP).

[0463] In one specific example, activation of the non-TGF-β mediated signaling pathway can decrease the expression level of E-cadherin.

[0464] In one specific example, activation of the non-TGF-β mediated signaling pathway may increase the expression level of any one or more of Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0465] In one embodiment, inhibition of the non-TGF-β mediated signaling pathway may result in inhibition of expression or a decrease in the expression level of any one or more of CSF3R, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin.

[0466] In one embodiment, inhibition of the non-TGF-β mediated signaling pathway may induce or increase the expression level of one or more of matrix metalloproteinase (MMP) and E-cadherin.

[0467] 2.5.4. Mechanism of CSF3R inhibitors to inhibit non-TGF-β-mediated signaling pathways

[0468] The CSF3R inhibitor of the present disclosure inhibits the CSF3-associated and non-TGF-β-mediated signaling pathways. That is, the CSF3R inhibitor can reduce the operation or activation of the CSF3R-mediated but not TGF-β-mediated signaling pathway involved in fibrosis.

[0469] In one specific example, during the CSF3-associated and non-TGF-β-mediated signaling pathway process, the TGF-β-mediated signaling pathway can be triggered (or activated). That is, a CSF3 inhibitor can inhibit the CSF3-associated and non-TGF-β-mediated signaling pathway, thereby inhibiting the TGF-β-mediated signaling pathway.

[0470] In one specific example, the CSF3R inhibitor can inhibit the CSF3-associated and non-TGF-β mediated signaling pathway by inhibiting the interaction (or binding) of CSF3R and CSF3.

[0471] In one specific example, the CSF3R inhibitor inhibits the CSF3-related and non-TGF-β mediated signaling pathways by inhibiting CSF3R expression or function, and may inhibit epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and / or extracellular matrix deposition. In another specific example, the CSF3R inhibitor may inhibit the interaction of CSF3R and CSF3, thereby inhibiting EMT, FMT, and / or extracellular matrix deposition.

[0472] In one specific example, the CSF3R inhibitor can inhibit the expression or reduce the expression level of any one or more of CSF3R, TGF-β, α-SMA, COL1A1, IL-11, collagen, fibronectin, hyaluronic acid, Versican, Osteopontin (OPN), hyaluronan synthase (HAS), tissue inhibitor of metalloproteinase (TIMP), Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin, the expression levels of which are increased due to activation of the non-TGF-β mediated signaling pathway.

[0473] In one specific example, the CSF3R inhibitor can induce or increase the expression level of one or more of matrix metalloproteinases (MMPs) and E-cadherin, the expression levels of which are reduced due to activation of the non-TGF-β mediated signaling pathway.

[0474] 2.6. Mechanism of action of CSF3R inhibitors 3: Inhibition of positive feedback loop

[0475] In one specific example, the CSF3R inhibitor may inhibit one or more of the following positive-feedback loops:

[0476] Positive feedback loop of CSF3R,

[0477] The positive feedback loop of TGF-β, and

[0478] A reciprocal positive feedback loop between CSF3R and TGF-β.

[0479] In another specific example, the CSF3R inhibitor can inhibit the positive feedback loop of CSF3R, the positive feedback loop of TGF-β, and / or the reciprocal positive feedback loop between CSF3R and TGF-β in the tissue (or cell) of the subject, i.e., reduce the action of the positive feedback loops.

[0480] 2.6.1. Implications of the positive feedback loop of CSF3R

[0481] The term “positive feedback loop of CSF3R” in the present disclosure refers to a signaling pathway in which an increase in CSF3R further induces the expression of CSF3R. In other words, the positive feedback loop of CSF3R refers to a process in which CSF3R causes more CSF3R to be produced, and the produced CSF3R causes more CSF3R to be produced again. In one specific example, activation of the positive feedback loop of CSF3R can induce the expression of CSF3R or increase the expression level of CSF3R. Therefore, the “positive feedback loop of CSF3R” in the present specification may also be expressed as “an auto-positive feedback loop of CSF3R.”

[0482] In one specific example, the positive feedback loop of CSF3R may be activated through one or more of the following processes:

[0483] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by CSF3R;

[0484] Increased expression of CSF3R and / or TGF-β by activated TGF-β-mediated signaling pathway and / or activated non-TGF-β-mediated signaling pathway;

[0485] Further activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3R;

[0486] TGF-β-mediated signaling pathways are further activated by expressed TGF-β; and

[0487] The expression levels of CSF3R and / or TGF-β are further increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway.

[0488] At this time, the process of “the expression level of CSF3R and / or TGF-β is increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway is activated by the expressed CSF3R; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression level of CSF3R is increased may be activated due to this repeating process (or loop structure).

[0489] 2.6.2. Implications of the positive feedback loop of TGF-β

[0490] The term “positive feedback loop of TGF-β” in the present disclosure refers to a signaling pathway in which an increase in TGF-β further induces the expression of TGF-β. In other words, the positive feedback loop of TGF-β refers to a process in which more TGF-β is produced due to TGF-β, and the produced TGF-β in turn produces more TGF-β. In one specific example, activation of the positive feedback loop of TGF-β can induce the expression of TGF-β or increase the expression level of TGF-β. Therefore, the “positive feedback loop of TGF-β” in the present specification may also be expressed as a “self-positive feedback loop of TGF-β.”

[0491] In one specific example, the positive-feedback loop of TGF-β can be activated through one or more of the following processes:

[0492] TGF-β-mediated signaling pathway is activated by TGF-β;

[0493] Increased expression of CSF3R and / or TGF-β by activated TGF-β-mediated signaling pathway;

[0494] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3R;

[0495] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0496] The expression levels of CSF3R and / or TGF-β are further increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway.

[0497] At this time, the process of “the expression amount of CSF3R and / or TGF-β is increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway is activated by the expressed CSF3R; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and a positive-feedback loop in which the expression amount of TGF-β increases may be activated due to this repeating process (or loop structure).

[0498] 2.6.3. Implications of the reciprocal positive feedback loop between CSF3R and TGF-β

[0499] The term “mutual positive-feedback loop between CSF3R and TGF-β” in the present disclosure refers to a reciprocal relationship between CSF3R and TGF-β that influences the increase in each other’s expression, and refers to a signaling pathway in which i) the expression of TGF-β is induced by an increase in CSF3R, and ii) the expression of CSF3R is induced by an increase in TGF-β. In other words, the mutual positive-feedback loop between CSF3R and TGF-β refers to a process in which more TGF-β is produced due to CSF3R, and the produced TGF-β in turn produces more CSF3R. Alternatively, the mutual positive-feedback loop between CSF3R and TGF-β refers to a process in which more CSF3R is produced due to TGF-β, and the produced CSF3R in turn produces more TGF-β.

[0500] Through experiments, the present inventors have elucidated a reciprocal positive feedback loop mechanism between CSF3R and TGF-β. Specifically, in Experimental Example 2.5, we confirmed that when cells were treated with CSF3 protein, the expression levels of CSF3, CSF3R, and TGF-β increased. Furthermore, we confirmed that the increased expression levels of CSF3, CSF3R, and TGF-β were reduced by a CSF3R inhibitor. These results suggest that the interaction between CSF3 and CSF3R increases the expression of CSF3, CSF3R, and TGF-β.

[0501] In one specific example, activation of the reciprocal positive-feedback loop between CSF3R and TGF-β can induce expression of CSF3R and TGF-β or increase the expression levels of CSF3R and TGF-β.

[0502] In one specific example, the reciprocal positive feedback loop between CSF3R and TGF-β can be activated through one or more of the following processes:

[0503] Activation of TGF-β-mediated signaling pathways and / or activation of CSF3-associated and non-TGF-β-mediated signaling pathways by CSF3R;

[0504] Increased expression of CSF3R and / or TGF-β by activated TGF-β-mediated signaling pathway and / or activated CSF3-associated and non-TGF-β-mediated signaling pathway;

[0505] Activation of TGF-β-mediated signaling pathways and / or CSF3-associated and non-TGF-β-mediated signaling pathways by expressed CSF3R;

[0506] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0507] Increased expression of CSF3R and TGF-β by activated TGF-β-mediated signaling pathway and / or activated CSF3-associated and non-TGF-β-mediated signaling pathway.

[0508] At this time, the process of “the expression levels of CSF3R and TGF-β are increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway are activated by the expressed CSF3R; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and due to this repeating process (or loop structure), a positive-feedback loop in which the expression levels of CSF3R and TGF-β are increased may be activated.

[0509] In another specific example, the reciprocal positive feedback loop between CSF3R and TGF-β can be activated through one or more of the following processes:

[0510] TGF-β-mediated signaling pathway is activated by TGF-β;

[0511] Increased expression of CSF3R and / or TGF-β by activated TGF-β-mediated signaling pathway;

[0512] Activation of TGF-β-mediated signaling pathway and / or non-TGF-β-mediated signaling pathway by expressed CSF3R;

[0513] Activation of the TGF-β-mediated signaling pathway by expressed TGF-β; and

[0514] Increased expression of CSF3R and TGF-β by activated TGF-β-mediated signaling pathway and / or activated non-TGF-β-mediated signaling pathway.

[0515] At this time, the process of “the expression levels of CSF3R and TGF-β are increased by the activated TGF-β-mediated signaling pathway and / or the activated non-TGF-β-mediated signaling pathway” and “the TGF-β-mediated signaling pathway and / or the non-TGF-β-mediated signaling pathway are activated by the expressed CSF3R; and the TGF-β-mediated signaling pathway is activated by the expressed TGF-β” may be repeated, and due to this repeating process (or loop structure), a positive-feedback loop in which the expression levels of CSF3R and TGF-β are increased may be activated.

[0516] 2.6.4. Positive-feedback loop inhibition mechanism of CSF3R inhibitors

[0517] The CSF3R inhibitor of the present disclosure may inhibit one or more of the positive feedback loops described above.

[0518] For example, both the positive feedback loop of CSF3R and the positive feedback loop of TGF-β expression can be inhibited. Similarly, the CSF3R inhibitor can inhibit the reciprocal positive feedback loop between CSF3R and TGF-β.

[0519] In one specific example, the CSF3R inhibitor can inhibit both the expression of CSF3R and the expression of TGF-β. In another specific example, the CSF3R inhibitor can reduce both the expression amount of CSF3R and the expression amount of TGF-β. That is, by inhibiting CSF3R, both the expression of CSF3R and the expression of TGF-β can be inhibited.

[0520] In the positive feedback loop of CSF3R; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3R and TGF-β, CSF3R and TGF-β are closely related. In this close relationship, (i) inhibiting CSF3R can significantly inhibit the positive feedback loop of CSF3R; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3R and TGF-β, whereas (ii) inhibiting only TGF-β makes it difficult to significantly inhibit the positive feedback loop of CSF3R; the positive feedback of TGF-β expression; and the mutual positive feedback loop between CSF3R and TGF-β. That is, compared to inhibiting TGF-β, inhibiting CSF3R can inhibit the positive feedback loop of CSF3R; the positive feedback of TGF-β expression; and the reciprocal positive feedback loop between CSF3R and TGF-β is inhibited to a stronger level.

[0521] For example, when TGF-β is inhibited, the expression levels of CSF3R and TGF-β are reduced, and when CSF3R is inhibited, the expression levels of CSF3R and TGF-β are reduced. In this case, the expression levels of CSF3R and TGF-β are further reduced by CSF3R inhibition compared to TGF-β inhibition.

[0522] This is because inhibition of CSF3R inhibits both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway within the positive-feedback loop of CSF3R; the positive-feedback of TGF-β expression; and the mutual positive-feedback loop between CSF3R and TGF-β, whereas inhibition of TGF-β inhibits only the TGF-β-mediated signaling pathway.

[0523] 2.7. A key regulator in the signaling pathway that leads to fibrosis: CSF3R

[0524] The present inventors have shown for the first time that inhibition of CSF3R inhibits TGF-β-mediated signaling pathways and non-TGF-β-mediated signaling pathways, whereas increase of CSF3R activates TGF-β-mediated signaling pathways and non-TGF-β-mediated signaling pathways. In other words, it has been revealed for the first time that CSF3R is a key regulator throughout the signaling pathways that induce fibrosis or tissue fibrosis. Based on the above explanation, the present inventors speculate that CSF3R is an upstream regulator of TGF-β, based on the experimental data showing that CSF3R is a key regulator in the induction of fibrosis.

[0525] For example, when looking at Experimental Examples 2.3 to 2.7, when comparing i) the CSF3R expression level that is reduced by inhibiting only the TGF-β-mediated signaling pathway and ii) the CSF3R expression level that is reduced by inhibiting both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway, it can be seen that the reduction in the latter (ii) is greater.

[0526] For example, when looking at Experimental Examples 2.3 to 2.7, when comparing the expression levels of α-SMA, Col1A1, CSF3, IL11, and CSF3R, etc., which are reduced by suppressing only the TGF-β-mediated signaling pathway and ii) the expression levels of α-SMA, Col1A1, CSF3, IL11, and CSF3R, etc., which are reduced by suppressing both the TGF-β-mediated signaling pathway and the non-TGF-β-mediated signaling pathway, it can be seen that the reduction in the latter (ii) is greater.

[0527] For example, looking at Experimental Example 2.5, Fig. 17, and Fig. 18, it can be seen that when the interaction between TGF-β and TGF-β receptor (TGF-βR) was inhibited, the expression levels of fibrosis-related markers were slightly reduced. In contrast, looking at Experimental Example 2.5, Experimental Example 2.6, Fig. 17, Fig. 18, and Fig. 19, it can be seen that when the interaction between CSF3 and CSF3 receptor (CSF3R) was inhibited, the expression levels of fibrosis-related markers were significantly reduced. That is, it can be seen that when the interaction between CSF3 and CSF3 receptor (CSF3R) was inhibited, the expression levels of fibrosis-related markers were closer to the control group (normal group in which fibrosis was not induced) than when the interaction between TGF-β and TGF-β receptor was inhibited.

[0528] 2.8. Effect of CSF3R inhibitors on returning fibrotic tissue to normal tissue

[0529] The CSF3R inhibitor of the present disclosure restores fibrotic tissue (or cells) to normal tissue (or cells). In another specific embodiment, the CSF3R inhibitor causes fibrotic tissue (or cells) to return to normal tissue (or cells).

[0530] Looking at the experimental examples of this specification and FIGS. 15, 20, 26, 28, 29, 41, 42, 43, and 45, it can be seen that fibrotic tissues (or cells) return to normal tissues (or cells) due to CSF3R inhibition.

[0531] In contrast, commercially available pulmonary fibrosis treatments (e.g., nintedanib or pirfenidone) that inhibit the TGF-β-mediated signaling pathway do not effectively restore fibrotic tissue (or cells) to normal tissue (or cells). In other words, it is widely recognized in the art that these treatments only alleviate fibrotic symptoms and do not restore normal tissue.

[0532] However, the CSF3R inhibitor disclosed herein has a therapeutic effect of restoring fibrotic tissue to normal tissue.

[0533] The reason for the difference in the effectiveness of commercialized pulmonary fibrosis treatments and the CSF3R inhibitors may be that, compared to commercialized pulmonary fibrosis treatments, CSF3R inhibitors not only inhibit the TGF-β-mediated signaling pathway, but also have a greater effect of inhibiting the non-TGF-β-mediated signaling pathway. From another perspective, the reason for the difference in the effectiveness of commercialized pulmonary fibrosis treatments and the CSF3R inhibitors may be that, compared to commercialized pulmonary fibrosis treatments, CSF3R inhibitors have a stronger effect of inhibiting the TGF-β-mediated signaling pathway.

[0534] 2.9. Examples of CSF3R inhibitors

[0535] The CSF3R inhibitor described in the present disclosure is not limited to anything that can inhibit the function of CSF3R. The CSF3R inhibitor may 1) interfere with the interaction of the CSF3R protein with other molecules, or 2) inhibit the expression of the CSF3R protein, thereby reducing the absolute amount of the CSF3R protein that interacts with other molecules, thereby resulting in the interference with the function of the CSF3R protein. Examples of CSF3R inhibitors capable of such actions include, but are not limited to, a nucleic acid that silences the mRNA of the CSF3R gene, a CRISPR / Cas system that knocks out or knocks down the CSF3R gene, a compound that inhibits the CSF3R protein, or an aptamer that inhibits the CSF3R protein.

[0536] 2.9.1. Nucleic acid that silences the mRNA of the CSF3R gene

[0537] The above CSF3R inhibitor may silence the mRNA of the CSF3R gene and thereby interfere with the translation process of the CSF3R protein. In one embodiment, the CSF3R inhibitor may be a nucleic acid that can specifically bind to mRNA transcribed from the CSF3R gene, and may be selected from siRNA (small interfering RNA), shRNA (small hairpin RNA), miRNA (microRNA), and antisense nucleic acid.

[0538] 2.9.2. CRISPR / Cas system targeting the CSF3R gene 1 - nuclease

[0539] The CSF3R inhibitor may knock out the CSF3R gene in the cellular genome, thereby preventing the production of the CSF3R protein. For example, the CRISPR / Cas system, which can act as a target-specific nuclease, can cleave a specific gene in the cellular genome, and can induce an error in the cellular gene repair mechanism (e.g., non-homologous end repair (NHEJ) and / or homologous recombination repair (HDR)) to generate an indel in the specific gene, thereby knocking out the specific gene. In one embodiment, the CSF3R inhibitor may be a composition comprising a CRISPR / Cas system, which comprises: a Cas protein or a nucleic acid encoding the same; and a guide RNA or a nucleic acid encoding the same, wherein the guide RNA can target the CSF3R gene in the cellular genome and interact with the Cas protein to form a complex.

[0540] 2.9.3. CRISPR / Cas system targeting the CSF3R gene 2 - regulator

[0541] The CSF3R inhibitor may suppress the production of the CSF3R protein by causing a knockdown effect that suppresses the expression of the CSF3R gene in the cellular genome. For example, in one target-specific embodiment, the CSF3R inhibitor may be a composition comprising a CRISPRi system comprising: a fusion protein in which a Cas protein and at least one gene expression regulatory domain are linked, or a nucleic acid encoding the fusion protein; and a guide RNA and a nucleic acid encoding the same, wherein the guide RNA can target the CSF3R gene in the cellular genome and interact with the fusion protein to form a complex. In this case, the gene expression regulatory domain may be a polypeptide or protein that suppresses the expression of the CSF3R gene in the cellular genome.

[0542] 2.9.4. Substance 1 that inhibits CSF3R protein - Compound

[0543] In one embodiment, the CSF3R inhibitor may be a compound and / or small molecule that functions to prevent the CSF3R protein from interacting with other molecules.

[0544] 2.9.5. Substance 2 that inhibits CSF3R protein - Aptamer

[0545] In one embodiment, the CSF3R inhibitor may be a DNA and / or RNA aptamer that functions to prevent the CSF3R protein from interacting with other molecules.

[0546] 2.9.6. Substance 3 that inhibits CSF3R protein - Antibodies

[0547] In one embodiment, the CSF3R inhibitor may be an anti-CSF3R antibody or a fragment of the anti-CSF3R antibody. The anti-CSF3R antibody refers to an antibody (or immunoglobulin) that recognizes the CSF3R protein as an antigen. The anti-CSF3R antibody refers to an antibody having a function of recognizing all or part of the CSF3R protein; and / or a function of binding to all or part of the CSF3R protein. The fragment of the anti-CSF3R antibody refers to a molecule that includes a part of the anti-CSF3R antibody and has the function of an anti-CSF3R antibody (a function of recognizing and binding to all or part of the CSF3R protein). That is, the anti-CSF3R antibody or the fragment of the anti-CSF3R antibody can recognize and / or bind to all or part of the CSF3R protein. Accordingly, the anti-CSF3R antibody or fragment thereof can bind to the CSF3R protein expressed in the tissue where fibrosis has occurred, thereby preventing the CSF3R protein from interacting with other molecules (e.g., CSF3), and consequently inhibiting the function of the CSF3R protein. The anti-CSF3R antibody or fragment thereof is not otherwise limited as long as it can inhibit CSF3R.

[0548] In one specific example, the anti-CSF3R antibody may be immunoglobulin G (IgG). In this case, the IgG may be selected from IgG1, IgG2, IgG3, and IgG4.

[0549] In one specific example, the anti-CSF3R antibody may be any one of a human antibody, a humanized antibody, and a chimeric antibody.

[0550] In one specific example, the anti-CSF3R antibody may be any one selected from a monoclonal antibody, a polyclonal antibody, a hybridoma monoclonal antibody, and a recombinant monoclonal antibody.

[0551] In one specific example, the anti-CSF3R antibody may be a human IgG.

[0552] In one specific example, the anti-CSF3R antibody may be a CSF3R neutralizing antibody.

[0553] In one specific example, the fragment of the anti-CSF3R antibody may be a fragment of a CSF3R neutralizing antibody.

[0554] In one specific example, the fragment of the anti-CSF3R antibody may be selected from F(ab), F(ab'), F(ab')2, monospecific F(ab')2, bispecific F(ab')2, scFv, ScFv-Fc, and sdAb.

[0555]

[0556] 3. Use of CSF3 inhibitors in the treatment of fibrosis

[0557] 3.1. Background - Therapeutic effect of CSF3 inhibitors on pulmonary fibrosis

[0558] Fibrosis can occur not only in the lungs but also in various tissues, including the liver and skin. However, the internal environment of the tissue where fibrosis occurs, the environment surrounding the tissue where fibrosis occurs, the intracellular environment of the tissue where fibrosis occurs, and the extracellular environment of the tissue where fibrosis occurs all differ, making it uncertain whether the same treatment strategy will work. Therefore, until confirmed through actual experiments, it is difficult to predict whether a CSF3 inhibitor that is effective in treating pulmonary fibrosis will also be effective in treating fibrosis in other tissues. For example, nintedanib and pirfenidone are used to treat pulmonary fibrosis, but not as treatments for fibrosis in other tissues.

[0559] 3.2. Overview of the use of CSF3 inhibitors in the treatment of fibrosis

[0560] In one aspect of the present disclosure, a CSF3 inhibitor is disclosed for use in the treatment of fibrosis. The CSF3 inhibitor of the present disclosure functions to directly and / or indirectly inhibit or interfere with the function of CSF3 in vivo. Specifically, the CSF3 inhibitor may inhibit the interaction of the CSF3 protein with other molecules or inhibit the expression of the CSF3 protein. The inventors of the present disclosure have discovered that when the function of CSF3 in a fibrotic tissue is inhibited, the fibrotic tissue is restored to normal tissue and fibrosis is treated. Through research, the mechanism of the CSF3 inhibitor for treating fibrosis has been elucidated.

[0561] The above CSF3 inhibitor has a therapeutic mechanism of inhibiting the induction of epithelial to mesenchymal transition (EMT) of tissue epithelial cells, inhibiting differentiation into myofibroblasts by suppressing the activity of fibroblasts, and / or reducing myofibroblasts by directly acting on myofibroblasts. The CSF3 inhibitor can inhibit a TGF-β-mediated signaling pathway; a non-TGF-β-mediated signaling pathway; a positive-feedback loop of CSF3; and / or a positive-feedback loop of TGF-β.

[0562] In one specific example, the CSF3 inhibitor can inhibit epithelial to mesenchymal transition (EMT) of tissue epithelial cells involving the TGF-β mediated signaling pathway, fibroblast-to-myofibroblast transdifferentiation (FMT) involving the TGF-β mediated signaling pathway, and / or extracellular matrix deposition involving the TGF-β mediated signaling pathway.

[0563] In one specific example, the CSF3 inhibitor can inhibit epithelial to mesenchymal transition (EMT) of tissue epithelial cells involving CSF3-related and non-TGF-β mediated signaling pathways, Fibroblast-to-Myofibroblast Transdifferentiation (FMT) of fibroblasts involving non-TGF-β mediated signaling pathways, and / or Extracellular Matrix deposition involving non-TGF-β mediated signaling pathways.

[0564] A detailed description of the inhibition of the TGF-β-mediated signaling pathway, non-TGF-β-mediated signaling pathway, positive-feedback loop of CSF3, and / or positive-feedback loop of TGF-β by the above CSF3 inhibitor is provided in the table of contents and its sub-tables of contents.

[0565] When the above CSF3 inhibitor is administered to tissue where fibrosis has occurred, the extracellular matrix accumulated in the fibrosis tissue is decomposed and myofibroblasts are reduced, resulting in a therapeutic effect where the fibrosis tissue returns to normal tissue.

[0566] In one specific example, when the CSF3 inhibitor is administered to a fibrotic tissue, 1) abnormally accumulated extracellular matrix, particularly accumulated collagen, is removed so that the extracellular matrix of the tissue returns to a normal state, and 2) abnormal myofibroblasts are reduced so that the tissue returns to a normal state. In one specific example, the CSF3 inhibitor treats fibrosis by 1) decomposing extracellular matrix derived from myofibroblasts accumulated in the fibrotic tissue, 2) preventing myofibroblasts in the fibrotic tissue from secreting extracellular matrix, and 3) reducing myofibroblasts in the fibrotic tissue. More specifically, the CSF3 inhibitor may have a therapeutic mechanism of directly acting on myofibroblasts to inactivate the myofibroblasts. The inactivation of the myofibroblasts may include 1) an extracellular matrix remodeling mechanism, 2) a mechanism of inhibiting extracellular matrix production by myofibroblasts, and 3) a mechanism of reducing myofibroblasts in the fibrotic tissue. The above CSF3 inhibitor can inactivate and / or induce inactivation of myofibroblasts. The therapeutic effects and therapeutic mechanisms are described in more detail in the table of contents below.

[0567] 3.3. Significance of CSF3 inhibitors

[0568] The CSF3 inhibitor of the present disclosure refers to a substance that ultimately inhibits the function of CSF3. The type, form, and composition of the CSF3 inhibitor are not otherwise limited as long as it can inhibit the function of CSF3. Specifically, inhibiting the function of CSF3 means 1) binding to the CSF3 protein to inhibit the function of the protein itself, 2) inhibiting the interaction of the CSF3 protein with other molecules, 3) inhibiting the expression of CSF3, thereby blocking the opportunity for CSF3 to function, or 4) inhibiting the function of CSF3 through other direct or indirect pathways.

[0569] In one specific example, the CSF3 inhibitor may be an anti-CSF3 antibody. In another specific example, the CSF3 inhibitor may be a nucleic acid molecule capable of silencing the mRNA of the CSF3 gene or causing RNA interference. Specifically, the nucleic acid molecule may include an antisense oligonucleotide; an interfering RNA (iRNA), such as miRNA, siRNA (small interfering RNA), or shRNA (short hairpin RNA). In another specific example, the CSF3 inhibitor may be a compound that inhibits the function of CSF3.

[0570] 3.4. CSF3 inhibitors' anti-fibrotic mechanism 1 - extracellular matrix degradation

[0571] 3.4.1. Overview of extracellular matrix degradation

[0572] The CSF3 inhibitor described above can restore fibrotic tissue to normal tissue. Specifically, the CSF3 inhibitor can decompose or remove the extracellular matrix secreted and accumulated by myofibroblasts in fibrotic tissue (or fibrotic tissue). At this time, the CSF3 inhibitor decomposes or removes the accumulated extracellular matrix, thereby restoring the extracellular matrix environment of the fibrotic tissue to that of a normal tissue. In the present disclosure, the therapeutic mechanism is referred to as "extracellular matrix degradation."

[0573] The extracellular matrix accumulated within the tissue where the above fibrosis has occurred may include collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN). The accumulation of the extracellular matrix interferes with the normal functioning of the tissue and may cause various pathological symptoms. In one embodiment, the degradation of the extracellular matrix may be the degradation of collagen accumulated within the fibrotic tissue.

[0574] When the CSF3 inhibitor is administered to a tissue where fibrosis has occurred, the extracellular matrix secreted by myofibroblasts is removed through various pathways. In one embodiment, the CSF3 inhibitor can promote the secretion and / or activation of matrix metalloproteinase (MMP) in the tissue. In another embodiment, the CSF3 inhibitor can inhibit the secretion and / or activation of tissue inhibitor of metalloproteinase (TIMP) in the tissue. In another embodiment, the CSF3 inhibitor can inhibit the secretion and / or activation of hyaluronan synthase 3 (HAS3) in the tissue.

[0575] 3.4.2. Example 1 of the extracellular matrix degradation pathway - MMP secretion promotion

[0576] Matrix metalloproteinases (MMPs) are a type of proteinase, also known as matrixin, and are enzymes involved in the degradation of the extracellular matrix. Therefore, when MMPs are secreted and activated within tissues, accumulated extracellular matrix, particularly collagen, is degraded. In one embodiment, the CSF3 inhibitor may promote the secretion and / or activation of MMPs within fibrotic tissues. Specifically, the MMPs may be at least one selected from MMP2, MMP9, and MMP13.

[0577] 3.4.3. Example 2 of the extracellular matrix degradation pathway - TIMP secretion inhibition

[0578] As the name suggests, tissue inhibitor of matrix metalloproteinase (TIMP) is a protein that inhibits the activity of matrix metalloproteinase (MMP). TIMP regulates the activity of the aforementioned MMP, and more specifically, it inhibits the activity of MMP. Therefore, when the secretion and / or activation of TIMP within tissues is inhibited, the activity of MMP increases, thereby allowing the degradation of accumulated extracellular matrix.

[0579] 3.5. CSF3 inhibitors' antifibrotic therapeutic mechanism 2 - Inhibition of extracellular matrix production by myofibroblasts

[0580] 3.5.1. Overview of inhibition of extracellular matrix secretion

[0581] The CSF3 inhibitor can inhibit the production and secretion of extracellular matrix by myofibroblasts in fibrotic tissue (or fibrotic tissue). This prevents the extracellular matrix secreted by the myofibroblasts from accumulating further in the fibrotic tissue. Here, the extracellular matrix may be collagen, fibronectin, hyaluronic acid, versican, and / or osteopontin (OPN). In order to prevent further accumulation of extracellular matrix in the fibrotic tissue, the production and secretion of extracellular matrix by the myofibroblasts must be inhibited. When the CSF3 inhibitor is administered to the fibrotic tissue, the production and secretion of extracellular matrix by the myofibroblasts is inhibited through various routes. In one embodiment, the CSF3 inhibitor can inhibit the expression and secretion of collagen and / or osteopontin (OPN) by the myofibroblasts in the fibrotic tissue. In another embodiment, the CSF3 inhibitor can inhibit the production of hyaluronan synthase (HAS) by myofibroblasts in fibrotic tissue, thereby inhibiting the synthesis and secretion of hyaluronic acid. This works in conjunction with the therapeutic mechanism 1 to remove abnormally accumulated extracellular matrix in fibrotic tissue and restore it to a normal state.

[0582] 3.5.2. Example 1 of the extracellular matrix secretion inhibition pathway - Inhibition of collagen expression

[0583] The above CSF3 inhibitor inhibits collagen expression. Collagen is one of the most actively secreted extracellular matrices from myofibroblasts in fibrotic tissues. Therefore, blocking the expression and secretion of collagen by myofibroblasts is as important as removing the accumulated collagen. The above CSF3 inhibitor can inhibit the expression and secretion of collagen by myofibroblasts in fibrotic tissues. As shown in Experimental Example 3, when a CSF3 inhibitor is administered to fibrotic tissues and / or cells, collagen expression in myofibroblasts is reduced.

[0584] 3.5.3. Example 2 of the extracellular matrix secretion inhibition pathway - HAS expression inhibition

[0585] The above CSF3 inhibitor inhibits the expression of hyaluronic acid synthase. Hyaluronan synthase (HAS) is an enzyme that synthesizes hyaluronic acid and is involved in the synthesis of hyaluronic acid in the cell plasma membrane. Therefore, when the expression of HAS within a cell is suppressed, the synthesis and secretion of hyaluronic acid are suppressed, thereby inhibiting the production of hyaluronic acid by myofibroblasts. The HAS may be HAS1, HAS2, and / or HAS3.

[0586] 3.5.4. Example 3 of the extracellular matrix secretion inhibition pathway - OPN expression inhibition

[0587] The above CSF3 inhibitor suppresses the expression of osteopontin. Osteopontin (OPN) is a major phosphoprotein that constitutes the skeleton and is involved in bone remodeling. Furthermore, OPN is known to be associated with the onset and progression of tissue fibrosis. The above CSF3 inhibitor can suppress the expression and secretion of OPN by myofibroblasts in fibrotic tissues.

[0588] 3.6. Mechanism of CSF3 Inhibitors to Treat Fibrosis 3 - Reduction of Myofibroblasts in Fibrotic Tissue

[0589] 3.6.1. Overview of the Reduction of Myofibroblasts in Fibrotic Tissue

[0590] The above CSF3 inhibitor can reduce myofibroblasts in fibrotic tissue (or fibrotic tissue). The reduction in myofibroblasts may be the result of one or more of: 1) apoptosis of myofibroblasts in fibrotic tissue, 2) inhibition of differentiation of epithelial cells and / or fibroblasts in fibrotic tissue into myofibroblasts, and 3) reversion of myofibroblasts in fibrotic tissue into tissue epithelial cells and / or fibroblasts. This allows fibrotic tissue to be restored to normal tissue. This effect of reducing myofibroblasts in fibrotic tissue is well demonstrated in Experimental Example 3, etc.

[0591] 3.6.2. Reduction of myofibroblasts due to apoptosis

[0592] As described above, when administered into fibrotic tissue, the CSF3 inhibitor inactivates myofibroblasts present within the fibrotic tissue. As a result, the myofibroblasts themselves undergo apoptosis, resulting in a decrease in the number of myofibroblasts.

[0593] 3.6.3. Inhibition of epithelial-mesenchymal transition of epithelial cells within tissues

[0594] The above CSF3 inhibitor can also inhibit epithelial-to-mesenchymal transition (EMT), which is the reverse differentiation of normal epithelial cells in fibrotic tissues into mesenchymal cells. As described above, through the epithelial-to-mesenchymal transition, normal epithelial cells are reverse differentiated into mesenchymal cells and then differentiate into myofibroblasts. Therefore, when the epithelial-to-mesenchymal transition is inhibited, the number of myofibroblasts no longer increases.

[0595] 3.6.4. Inhibition of differentiation of fibroblasts into myofibroblasts within the tissue

[0596] The above CSF3 inhibitor can also inhibit the differentiation of fibroblasts in fibrotic tissue into myofibroblasts. As described above, fibroblasts differentiate into myofibroblasts during the progression of fibrosis. Therefore, inhibiting the differentiation of fibroblasts into myofibroblasts prevents further growth of myofibroblasts.

[0597] 3.6.5. Reversion of myofibroblasts to tissue epithelial cells

[0598] The above CSF3 inhibitor can induce or promote the reversion of myofibroblasts back to epithelial cells. As described above, just as epithelial cells can transform into myofibroblasts through epithelial-mesenchymal transition, the reverse process of epithelial-mesenchymal transition can occur, allowing myofibroblasts to revert to epithelial cells in normal tissues. This can work in conjunction with the reduction of myofibroblasts due to apoptosis and the inhibition of the dedifferentiation of epithelial cells into myofibroblasts. The results of Experimental Example 3 provide indirect evidence that myofibroblasts revert to tissue epithelial cells.

[0599] 3.6.6. Reversion of myofibroblasts to fibroblasts

[0600] The above CSF3 inhibitor can induce or promote the reversion of myofibroblasts back to fibroblasts. Just as fibroblasts can differentiate into myofibroblasts as described above, the reverse process of differentiation can occur, allowing myofibroblasts to revert back to fibroblasts. This can work in conjunction with the reduction of myofibroblasts due to apoptosis and the inhibition of fibroblast differentiation into myofibroblasts.

[0601] 3.6.7. Biomarkers associated with inhibition of epithelial-mesenchymal transition, or the reversion of myofibroblasts to tissue epithelial cells and / or fibroblasts.

[0602] Biomarkers associated with epithelial-mesenchymal transition (EMT), or the reversion of myofibroblasts to tissue epithelial cells, include E-cadherin, Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin. Specifically, when EMT occurs, the expression level of E-cadherin may decrease, and / or the expression level of one or more markers selected from Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin may increase. Conversely, when the expression level of E-cadherin increases, and / or the expression level of one or more markers selected from Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin decreases, it can be interpreted that 1) EMT is inhibited and differentiation into myofibroblasts no longer occurs, or 2) myofibroblasts revert to tissue epithelial cells.

[0603] 3.7. Advantages of CSF3 inhibitors for fibrosis treatment 1 - Fibrosis treatment effect

[0604] When the anti-fibrotic agent containing the CSF3 inhibitor is administered to a tissue with fibrosis, it has the effects of 1) decomposing accumulated myofibroblast-derived extracellular matrix, particularly collagen, and 2) reducing myofibroblasts within the tissue, thereby restoring it to normal tissue. This means that the anti-fibrotic agent containing the CSF3 inhibitor has a direct and complete anti-fibrotic effect.

[0605] The inventors of the present invention have experimentally confirmed that accumulated extracellular matrix within fibrotic tissues is degraded, and the fibrotic tissues return to normal. Compared to conventional anti-fibrotic agents, which have only incomplete therapeutic effects, the anti-fibrotic agent comprising a CSF3 inhibitor exhibits complete therapeutic effects compared to conventional anti-fibrotic agents, making it a superior treatment.

[0606] The reason why anti-fibrotic agents containing CSF3 inhibitors have a complete therapeutic effect compared to conventional anti-fibrotic agents may be because the degree to which anti-fibrotic agents containing CSF3 inhibitors inhibit the TGF-β-mediated signaling pathway is superior to that of conventional anti-fibrotic agents. The reason why anti-fibrotic agents containing CSF3 inhibitors have a complete therapeutic effect compared to conventional anti-fibrotic agents may be because conventional anti-fibrotic agents only inhibit the TGF-β-mediated signaling pathway, whereas anti-fibrotic agents containing CSF3 inhibitors can inhibit non-TGF-β-mediated signaling pathways; the positive feedback loop of CSF3; and the positive feedback loop of TGF-β in addition to the TGF-β-mediated signaling pathway.

[0607] 3.8. Advantage 2 of fibrosis treatment using CSF3 inhibitors - Therapeutic effect regardless of the type of tissue where fibrosis occurs

[0608] The present inventors have revealed that CSF3 functions as a key regulator in the development of pulmonary fibrosis. Accordingly, pulmonary fibrosis treatments containing CSF3 inhibitors have a more fundamental mechanism of action than conventional pulmonary fibrosis treatments and exhibit true therapeutic effects. Furthermore, the present inventors experimentally demonstrated that CSF3 is also crucially involved in fibrosis in skin, liver, and kidney cells. These findings reasonably suggest that CSF3 is a key regulator (or upstream regulator) in tissue fibrosis, regardless of tissue type. Furthermore, this provides the basis for predicting that CSF3 inhibitors may have therapeutic effects on fibrosis regardless of tissue type.

[0609] Specifically, the inventors of the present disclosure have revealed the pathogenesis and / or treatment mechanism of fibrosis through Experimental Example 2, and have confirmed through Experimental Example 3 that a CSF3 inhibitor has a therapeutic effect on fibrosis occurring in various tissues. For example, Experimental Examples 3.1 and 3.2 confirmed that a CSF3 inhibitor has a therapeutic effect on pulmonary fibrosis. For example, Experimental Examples 3.3 and 3.4 confirmed that a CSF3 inhibitor has a therapeutic effect on skin fibrosis. For example, Experimental Example 3.5 confirmed that a CSF3 inhibitor has a therapeutic effect on liver fibrosis. For example, Experimental Example 3.6 confirmed that a CSF3 inhibitor has a therapeutic effect on renal fibrosis.

[0610] Therefore, the anti-fibrosis treatment agent including the CSF3 inhibitor disclosed herein can act as an effective treatment agent regardless of the type of tissue in which fibrosis occurs.

[0611] In one specific example, the tissue in which the fibrosis occurs may be any one tissue selected from the group consisting of skin; brain; nervous system; heart; bone marrow; liver; gut; joint; eye; lung; kidney; retroperitoneum; mediastinum; and pancreas.

[0612] In one specific example, the CSF3 inhibitor has a therapeutic effect on skin fibrosis.

[0613] In one specific example, the CSF3 inhibitor has a therapeutic effect on cerebral fibrosis.

[0614] In one specific example, the CSF3 inhibitor has a therapeutic effect on neurofibromatosis.

[0615] In one specific example, the CSF3 inhibitor has a therapeutic effect on cardiac fibrosis.

[0616] In one specific example, the CSF3 inhibitor has a therapeutic effect on myelofibrosis.

[0617] In one specific example, the CSF3 inhibitor has a therapeutic effect on liver fibrosis.

[0618] In one specific example, the CSF3 inhibitor has a therapeutic effect on intestinal fibrosis.

[0619] In one specific example, the CSF3 inhibitor has a therapeutic effect on arthrofibrosis.

[0620] In one specific example, the CSF3 inhibitor has an anti-fibrotic effect.

[0621] In one specific example, the CSF3 inhibitor has an effect in treating optic fibrosis.

[0622] In one specific example, the CSF3 inhibitor has a therapeutic effect on pulmonary fibrosis.

[0623] In one specific example, the CSF3 inhibitor has a therapeutic effect on renal fibrosis.

[0624] In one specific example, the CSF3 inhibitor has a therapeutic effect on retroperitoneal fibrosis.

[0625] In one specific example, the CSF3 inhibitor has a therapeutic effect on mediastinal fibrosis.

[0626] In one specific example, the CSF3 inhibitor has a therapeutic effect on pancreatic fibrosis.

[0627] 3.9. Pharmaceutical composition for treating fibrosis comprising a CSF3 inhibitor

[0628] 3.9.1. Overview of pharmaceutical compositions for treating fibrosis comprising CSF3 inhibitors

[0629] The present disclosure discloses a pharmaceutical composition for treating fibrosis comprising a CSF3 inhibitor. The pharmaceutical composition for treating fibrosis comprises a therapeutically effective amount of a CSF3 inhibitor. In one embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3 inhibitor and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3 inhibitor and an adjuvant. In another embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3 inhibitor; a pharmaceutically acceptable carrier; and an adjuvant. Here, the pharmaceutically acceptable carrier may be used for appropriate formulation of the pharmaceutical composition for treating fibrosis.

[0630] 3.9.2. Target disease - Fibrosis

[0631] The pharmaceutical composition for treating fibrosis is intended to treat fibrosis, or a disease, condition, and / or symptom associated with fibrosis. The fibrosis or a disease, condition, and / or symptom associated with fibrosis includes all those recognizable to a person skilled in the art.

[0632] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0633] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0634] 3.9.3. Examples of formulations of pharmaceutical compositions for treating fibrosis

[0635] In one specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor may be formulated as a troche, lozenge, tablet, aqueous suspension, oily suspension, prepared powder, granules, emulsion, hard capsule, soft capsule, syrup, or elixir. In another specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor may be formulated as an injection, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, or cream. In another specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor may be formulated as an injection. Specifically, a therapeutically effective amount of the CSF3 inhibitor may be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which may be formulated for unit dose in ampoules or vials. In another specific example, the pharmaceutical composition for treating fibrosis containing the CSF3 inhibitor can be formulated as an aerosol by mixing a propellant and other additives to prepare a water-dispersed concentrate or wet powder. In another specific example, when the pharmaceutical composition for treating fibrosis containing the CSF3 inhibitor is formulated for transdermal use, an ointment, cream, powder for application, oil, external skin preparation, etc. can be prepared by adding animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as a carrier to a therapeutically effective amount of the CSF3 inhibitor.

[0636] 3.9.4. Examples of pharmaceutically acceptable carriers

[0637] Pharmaceutically acceptable carriers are those commonly used in formulations, and include, but are not limited to, saline solution, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc., and may further include other conventional additives such as antioxidants and buffers as needed. In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, each ingredient can be preferably formulated using the methods disclosed in Remington's Pharmaceutical Sciences (19th edition, 1995). In one embodiment, the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor comprises a pharmaceutically acceptable carrier, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; an excipient such as dicalcium phosphate; a disintegrant such as corn starch or sweet potato starch; a lubricant such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; a sweetener; a fragrance; a syrup; a liquid carrier such as a fatty oil; a sterile aqueous solution; propylene glycol; polyethylene glycol; an injectable ester such as ethyl oleate; a suspension; an emulsion; a freeze-dried preparation; an external preparation; a stabilizer; a buffer; an animal oil; a vegetable oil; a wax; a paraffin; a starch; a tragacanth; a cellulose derivative; a polyethylene glycol; a silicone; a bentonite; a silica; talc; zinc oxide, or a suitable combination thereof.

[0638] 3.10. Treatment of fibrosis using CSF3 inhibitors

[0639] 3.10.1. Overview of Fibrosis Treatment Using CSF3 Inhibitors

[0640] The present disclosure discloses a method for treating fibrosis using the CSF3 inhibitor. The method for treating fibrosis comprises administering a therapeutically effective amount of the CSF3 inhibitor to a subject using an appropriate administration method, an appropriate administration regimen, and an appropriate dosage. In one specific example, the method for treating fibrosis comprises administering a therapeutically effective CSF3 inhibitor in an appropriate formulation, i.e., a pharmaceutical composition for treating fibrosis, to a subject using an appropriate administration method, an appropriate administration regimen, and an appropriate dosage. In one specific example, the subject of the method for treating fibrosis may be a human or a non-human animal. In one specific example, the subject of the method for treating fibrosis may be a subject suffering from fibrosis, or a fibrosis-related disease, condition, and / or symptom.

[0641] 3.10.2. Diseases to be treated - Fibrosis

[0642] The above fibrosis treatment method is intended to treat fibrosis, or a fibrosis-related disease, condition, and / or symptom. The fibrosis or fibrosis-related disease, condition, and / or symptom includes all those recognizable to a person skilled in the art.

[0643] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0644] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0645] 3.10.3. Examples of Dosage Methods

[0646] The above-mentioned method for treating fibrosis may involve administering a therapeutically effective CSF3 inhibitor, appropriately formulated, to a subject via an appropriate administration method. In one embodiment, the above-mentioned method for treating fibrosis may involve administering a therapeutically effective CSF3 inhibitor, appropriately formulated, orally or parenterally. The parenteral administration method may include intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, and subcutaneous administration.

[0647] Various criteria related to administration (dosage, cycle, frequency, etc.) can be adjusted by the physician according to the individual patient's needs based on clinical examination during the administration process.

[0648] 3.10.4. Example of Dosage

[0649] The above-mentioned method for treating fibrosis may involve administering to a subject an appropriate dosage of a therapeutically effective CSF3 inhibitor, appropriately formulated. In one embodiment, the dosage may be about 0.01 mg to 1000 mg per kg of the subject's body weight, based on the CSF3 inhibitor.

[0650] 3.10.5. Example of Dosage Cycle

[0651] The above method for treating fibrosis may be to administer to a subject an appropriately formulated therapeutically effective CSF3 inhibitor at an appropriate dosing cycle. In one embodiment, the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor at the appropriate dosage may be administered once a day. In another example, the dosing cycle may be to administer the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor at the appropriate dosage in divided doses at least twice a day. In another example, the dosing cycle may be to administer the pharmaceutical composition for treating fibrosis comprising the CSF3 inhibitor at the appropriate dosage at intervals of 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 1 week, 2 weeks, 1 month, and / or 3 months.

[0652] 3.11. Therapeutic Uses of CSF3 Inhibitors

[0653] The present disclosure discloses the use of a CSF3 inhibitor for the treatment of fibrosis. In one embodiment, the present disclosure discloses a CSF3 inhibitor for the treatment of fibrosis, or a fibrosis-related disease, condition, and / or symptom.

[0654] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0655] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0656] 3.12. Use for the production of therapeutic agents using CSF3 inhibitors

[0657] The present disclosure discloses the use of a CSF3 inhibitor for the preparation of a fibrosis treatment agent. In one specific example, the present disclosure discloses the use of a CSF3 inhibitor for the preparation of a medicament for the treatment of fibrosis, or a fibrosis-related disease, condition, and / or symptom.

[0658] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0659] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0660] 3.13. Examples of tissues where fibrosis occurs

[0661] Fibrosis can develop in virtually any tissue in the body, and it is known that many tissues within the body can become fibrotic, leading to a variety of diseases, conditions, and / or conditions. For example, fibrosis has been reported to occur in tissues such as the skin, brain, nervous system, heart, bone marrow, liver, gut, joints, eyes, lungs, kidneys, retroperitoneum, mediastinum, and / or pancreas.

[0662] 3.14. Examples of fibrotic diseases

[0663] The fibrosis or fibrosis-associated diseases, conditions, and / or symptoms described herein include those recognizable to a person skilled in the art. In one embodiment, the fibrosis or fibrosis-associated diseases, conditions, and / or symptoms include hypertrophic scar; systemic sclerosis; multiple cancers; pulmonary arterial hypertension; glial scar; Alzheimer's; cardiac fibrosis; hypertrophic cardiomyopathy; cardiac dysfunction; valvular disease; arrhythmia; myelofibrosis; myelodysplastic syndrome; chronic myelogenous leukemia; cirrhosis; portal hypertension; hepatocellular carcinoma; Nonalcoholic steatohepatitis (NASH); intestinal fibrosis; enteropathies; inflammatory bowel disease; arthrofibrosis; subretinal fibrosis; epithelial fibrosis; vision loss; idiopathic pulmonary fibrosis; cystic fibrosis; pulmonary hypertension; thromboembolic disease; emphysema;Renal fibrosis; cystic fibrosis; nephrogenic systemic fibrosis; chronic kidney disease; renal anemia; retroperitoneal fibrosis; mediastinal fibrosis; pancreatic fibrosis; cystic fibrosis; chronic pancreatitis; biliary duct obstruction; autoimmune-interstitial lung disease, including ankylosing spondylitis and rheumatoid arthritis; connective tissue disease-interstitial lung disease, including rheumatoid arthritis-interstitial lung disease; and non-idiopathic pulmonary fibrosis-interstitial lung disease.;

[0664] 3.15. Examples of CSF3 inhibitors

[0665] The CSF3 inhibitor described in the present disclosure is not limited to any other inhibitor that can inhibit the function of CSF3. As described above, the CSF3 inhibitor may 1) interfere with the interaction of the CSF3 protein with other molecules, or 2) inhibit the expression of the CSF3 protein, thereby reducing the absolute amount of the CSF3 protein that interacts with other molecules, thereby resulting in the interference with the function of the CSF3 protein. Examples of CSF3 inhibitors that can have such an effect include, but are not limited to, a nucleic acid that silences the mRNA of the CSF3 gene, a CRISPR / Cas system that knocks out or knocks down the CSF3 gene, a compound that inhibits the CSF3 protein, or an aptamer that inhibits the CSF3 protein.

[0666] As a specific example, the CSF3 inhibitor may be one described in the table of contents <1.9. Examples of CSF3 inhibitors> and its sub-tables <1.9.1. Nucleic acid that silences mRNA of CSF3 gene> to <1.9.6. Substance 3 that inhibits CSF3 protein - Antibody>.

[0667] In one specific example, the anti-CSF3 antibody may be one described in the table of contents under <5. Anti-CSF3 Antibody>.

[0668]

[0669] 4. Use of CSF3R inhibitors in the treatment of fibrosis

[0670] 4.1. Background - Therapeutic effect of CSF3R inhibitors on pulmonary fibrosis

[0671] Fibrosis can occur not only in the lungs but also in various tissues, including the liver and skin. However, the internal environment of the tissue where fibrosis occurs, the environment surrounding the tissue where fibrosis occurs, the intracellular environment of the tissue where fibrosis occurs, and the extracellular environment of the tissue where fibrosis occurs are all different, making it uncertain whether the same treatment strategy will work. Therefore, until confirmed through actual experiments, it is difficult to predict whether a CSF3R inhibitor that is effective in treating pulmonary fibrosis will also be effective in treating fibrosis in other tissues. For example, nintedanib and pirfenidone are used to treat pulmonary fibrosis, but not as treatments for fibrosis in other tissues.

[0672] 4.2. Overview of the Uses of CSF3R Inhibitors in Fibrosis Treatment

[0673] In one aspect of the present disclosure, a CSF3R inhibitor is disclosed for use in the treatment of fibrosis. The CSF3R inhibitor of the present disclosure functions to directly and / or indirectly inhibit or interfere with the function of CSF3R in vivo. Specifically, the CSF3R inhibitor may inhibit the interaction of the CSF3R protein with other molecules or inhibit the expression of the CSF3R protein.

[0674] The inventors of the present disclosure have confirmed that when a CSF3R inhibitor is treated in tissues with pulmonary fibrosis, the tissues return to normal tissue, and this effect is identical / similar to the effect of treating a CSF3R inhibitor in tissues with pulmonary fibrosis. It is a well-known fact that CSF3 and CSF3R have an interactive relationship, and as the inventors of the present disclosure have discovered, the effect observed when CSF3 is inhibited in pulmonary fibrosis is also observed when CSF3R is inhibited, so CSF3R inhibitors have a fibrosis treatment effect in all tissues like CSF3 inhibitors. Accordingly, the inventors of the present disclosure have conducted additional research on CSF3R, and have discovered that when the function of CSF3R is inhibited in tissues with fibrosis, the fibrotic tissue is restored to normal tissue and fibrosis is treated, and the fibrosis treatment mechanism of CSF3R inhibitors has been elucidated.

[0675] The above CSF3R inhibitor has a therapeutic mechanism that inhibits the induction of epithelial to mesenchymal transition (EMT) of tissue epithelial cells, inhibits differentiation into myofibroblasts by suppressing the activity of fibroblasts, and / or acts directly on myofibroblasts to reduce myofibroblasts. The CSF3R inhibitor can inhibit a TGF-β-mediated signaling pathway; a non-TGF-β-mediated signaling pathway; a positive-feedback loop of CSF3; and / or a positive-feedback loop of TGF-β. In one specific example, the CSF3R inhibitor can inhibit epithelial to mesenchymal transition (EMT) of tissue epithelial cells involving the TGF-β mediated signaling pathway, fibroblast-to-myofibroblast transdifferentiation (FMT) involving the TGF-β mediated signaling pathway, and / or extracellular matrix deposition involving the TGF-β mediated signaling pathway. In one specific example, the CSF3R inhibitor can inhibit epithelial to mesenchymal transition (EMT) of tissue epithelial cells involving a non-TGF-β mediated signaling pathway, Fibroblast-to-Myofibroblast Transdifferentiation (FMT) of fibroblasts involving a non-TGF-β mediated signaling pathway, and / or Extracellular Matrix deposition involving a non-TGF-β mediated signaling pathway. A detailed description of the inhibition of the TGF-β mediated signaling pathway; the non-TGF-β mediated signaling pathway; the positive-feedback loop of CSF3R; and / or the positive-feedback loop of TGF-β by the CSF3R inhibitor is <2.The mechanism of action of CSF3R inhibitors that cause fibrotic tissue to return to normal tissue is explained in the table of contents and its sub-tables.

[0676] When the CSF3R inhibitor is administered to a tissue where fibrosis has occurred, the extracellular matrix accumulated in the fibrosis has been decomposed and myofibroblasts have been reduced, thereby exhibiting a therapeutic effect in which the fibrosis has returned to a normal tissue. In one specific example, when the CSF3R inhibitor is administered to a tissue where fibrosis has occurred, 1) the abnormally accumulated extracellular matrix, particularly the accumulated collagen, has been removed, thereby restoring the extracellular matrix of the tissue to a normal state, and 2) abnormal myofibroblasts have been reduced, thereby restoring the tissue to a normal state. In one specific example, the CSF3R inhibitor treats fibrosis by 1) decomposing the extracellular matrix derived from myofibroblasts accumulated in the fibrosis has occurred, 2) preventing the myofibroblasts in the fibrosis has occurred from secreting the extracellular matrix, and 3) reducing the myofibroblasts in the fibrosis has occurred. More specifically, the CSF3R inhibitor may have a therapeutic mechanism in which it directly acts on myofibroblasts to inactivate the myofibroblasts. The above inactivation of myofibroblasts may include 1) an extracellular matrix remodeling mechanism, 2) a mechanism of inhibiting extracellular matrix production by myofibroblasts, and 3) a mechanism of reducing myofibroblasts within fibrotic tissue. The CSF3R inhibitor may inactivate myofibroblasts and / or induce their inactivation. The above therapeutic effects and therapeutic mechanisms are described in more detail in the table of contents below.

[0677] 4.3. Significance of CSF3R inhibitors

[0678] The CSF3R inhibitor of the present disclosure refers to a substance that ultimately inhibits the function of CSF3R. The type, form, and composition of the CSF3R inhibitor are not otherwise limited as long as it can inhibit the function of CSF3R. Specifically, inhibiting the function of CSF3R means 1) binding to the CSF3R protein to inhibit the function of the protein itself, 2) inhibiting the interaction of the CSF3R protein with other molecules, 3) inhibiting the expression of CSF3R and consequently blocking the opportunity for CSF3R to function, or 4) inhibiting the function of CSF3R through other direct or indirect pathways.

[0679] In one specific example, the CSF3R inhibitor may be an anti-CSF3R antibody. In another specific example, the CSF3R inhibitor may be a nucleic acid molecule capable of silencing the mRNA of the CSF3R gene or causing RNA interference. Specifically, the nucleic acid molecule may include an antisense oligonucleotide; an interfering RNA (iRNA), such as miRNA, siRNA (small interfering RNA), or shRNA (short hairpin RNA). In another specific example, the CSF3R inhibitor may be a compound that inhibits the function of CSF3R.

[0680] 4.4. CSF3R inhibitors' anti-fibrotic mechanism 1 - extracellular matrix degradation

[0681] 4.4.1. Overview of extracellular matrix degradation

[0682] The CSF3R inhibitor described above can restore fibrotic tissue to normal tissue. Specifically, the CSF3R inhibitor can decompose or remove the extracellular matrix secreted and accumulated by myofibroblasts in fibrotic tissue (or fibrotic tissue). At this time, the CSF3R inhibitor can decompose or remove the accumulated extracellular matrix, thereby restoring the extracellular matrix environment of the fibrotic tissue to that of a normal tissue. In the present disclosure, the therapeutic mechanism is referred to as "extracellular matrix decomposition."

[0683] The extracellular matrix accumulated within the tissue where the above fibrosis has occurred may include collagen, fibronectin, hyaluronic acid, versican, and osteopontin (OPN). The accumulation of the extracellular matrix interferes with the normal functioning of the tissue and may cause various pathological symptoms. In one embodiment, the degradation of the extracellular matrix may be the degradation of collagen accumulated within the fibrotic tissue.

[0684] When the CSF3R inhibitor is administered to a tissue where fibrosis has occurred, the extracellular matrix secreted by myofibroblasts is removed through various pathways. In one embodiment, the CSF3R inhibitor can promote the secretion and / or activation of matrix metalloproteinase (MMP) in the tissue. In another embodiment, the CSF3R inhibitor can inhibit the secretion and / or activation of tissue inhibitor of metalloproteinase (TIMP) in the tissue. In another embodiment, the CSF3R inhibitor can inhibit the secretion and / or activation of hyaluronan synthase 3 (HAS3) in the tissue.

[0685] 4.4.2. Example 1 of the extracellular matrix degradation pathway - MMP secretion promotion

[0686] Matrix metalloproteinases (MMPs) are a type of proteinase, also known as matrixin, and are enzymes involved in the degradation of the extracellular matrix. Therefore, when MMPs are secreted and activated within tissues, accumulated extracellular matrix, particularly collagen, is degraded. In one embodiment, the CSF3R inhibitor may promote the secretion and / or activation of MMPs within fibrotic tissues. Specifically, the MMPs may be at least one selected from MMP2, MMP9, and MMP13.

[0687] 4.4.3. Example 2 of the extracellular matrix degradation pathway - TIMP secretion inhibition

[0688] As the name suggests, tissue inhibitor of matrix metalloproteinase (TIMP) is a protein that inhibits the activity of matrix metalloproteinase (MMP). TIMP regulates the activity of the aforementioned MMP, and more specifically, it inhibits the activity of MMP. Therefore, when the secretion and / or activation of TIMP within tissues is inhibited, the activity of MMP increases, thereby allowing the degradation of accumulated extracellular matrix.

[0689] 4.5. CSF3 inhibitors' anti-fibrotic mechanism 2 - Inhibition of extracellular matrix production by myofibroblasts

[0690] 4.5.1. Overview of inhibition of extracellular matrix secretion

[0691] The CSF3R inhibitor can inhibit the production and secretion of extracellular matrix by myofibroblasts in fibrotic tissue (or fibrotic tissue). This prevents the extracellular matrix secreted by the myofibroblasts from accumulating further in the fibrotic tissue. Here, the extracellular matrix may be collagen, fibronectin, hyaluronic acid, versican, and / or osteopontin (OPN). In order to prevent further accumulation of extracellular matrix in the fibrotic tissue, the production and secretion of extracellular matrix by the myofibroblasts must be inhibited. When the CSF3R inhibitor is administered to the fibrotic tissue, the production and secretion of extracellular matrix by the myofibroblasts is inhibited through various pathways. In one embodiment, the CSF3R inhibitor can inhibit the expression and secretion of collagen and / or osteopontin (OPN) by the myofibroblasts in the fibrotic tissue. In another embodiment, the CSF3R inhibitor can inhibit the production of hyaluronan synthase (HAS) by myofibroblasts in fibrotic tissues, thereby inhibiting the synthesis and secretion of hyaluronic acid. This works in conjunction with the therapeutic mechanism 1 to remove abnormally accumulated extracellular matrix in fibrotic tissues and restore them to a normal state.

[0692] 4.5.2. Example 1 of the extracellular matrix secretion inhibition pathway - Inhibition of collagen expression

[0693] The above CSF3R inhibitor inhibits collagen expression. Collagen is one of the extracellular matrices most actively secreted by myofibroblasts in fibrotic tissues. Therefore, blocking the expression and secretion of collagen by myofibroblasts is as important as removing the accumulated collagen. The above CSF3R inhibitor can inhibit the expression and secretion of collagen by myofibroblasts in fibrotic tissues. As shown in Experimental Example 3, when a CSF3R inhibitor is administered to fibrotic tissues and / or cells, collagen expression in myofibroblasts is reduced.

[0694] 4.5.3. Example 2 of the extracellular matrix secretion inhibition pathway - HAS expression inhibition

[0695] The above CSF3R inhibitor inhibits the expression of hyaluronic acid synthase. Hyaluronan synthase (HAS) is an enzyme that synthesizes hyaluronic acid and is involved in the synthesis of hyaluronic acid in the cell plasma membrane. Therefore, when the expression of HAS within a cell is suppressed, the synthesis and secretion of hyaluronic acid are suppressed, thereby inhibiting the production of hyaluronic acid by myofibroblasts. The HAS may be HAS1, HAS2, and / or HAS3.

[0696] 4.5.4. Example 3 of the extracellular matrix secretion inhibition pathway - OPN expression inhibition

[0697] The above CSF3R inhibitor suppresses the expression of osteopontin. Osteopontin (OPN) is a major phosphoprotein that constitutes the skeleton and is involved in bone remodeling. Furthermore, OPN is known to be associated with the onset and progression of tissue fibrosis. The above CSF3 inhibitor can suppress the expression and secretion of OPN by myofibroblasts in fibrotic tissues.

[0698] 4.6. CSF3 inhibitors' anti-fibrotic mechanism 3 - Reduction of myofibroblasts in fibrotic tissue

[0699] 4.6.1. Overview of the Reduction of Myofibroblasts in Fibrotic Tissue

[0700] The above CSF3R inhibitor can reduce myofibroblasts in fibrotic tissue (or fibrotic tissue). The reduction in myofibroblasts may be the result of one or more of: 1) apoptosis of myofibroblasts in fibrotic tissue, 2) inhibition of differentiation of epithelial cells and / or fibroblasts in fibrotic tissue into myofibroblasts, and 3) reversion of myofibroblasts in fibrotic tissue into tissue epithelial cells and / or fibroblasts. This allows fibrotic tissue to be restored to normal tissue. This effect of reducing myofibroblasts in fibrotic tissue is well demonstrated in Experimental Example 3, etc.

[0701] 4.6.2. Reduction of myofibroblasts due to apoptosis

[0702] As described above, when administered into fibrotic tissue, the CSF3R inhibitor inactivates myofibroblasts present within the fibrotic tissue. As a result, the myofibroblasts themselves undergo apoptosis, resulting in a decrease in the number of myofibroblasts.

[0703] 4.6.3. Inhibition of epithelial-mesenchymal transition of epithelial cells within tissues

[0704] The above CSF3R inhibitor can also inhibit epithelial to mesenchymal transition (EMT), which is the reverse differentiation of normal epithelial cells in fibrotic tissues into mesenchymal cells. As described above, through the epithelial to mesenchymal transition, normal epithelial cells are reverse differentiated into mesenchymal cells and then differentiate into myofibroblasts. Therefore, when the epithelial to mesenchymal transition is inhibited, the number of myofibroblasts no longer increases.

[0705] 4.6.4. Inhibition of differentiation of fibroblasts into myofibroblasts within the tissue

[0706] The above CSF3R inhibitor can also inhibit the differentiation of fibroblasts in fibrotic tissues into myofibroblasts. As described above, fibroblasts differentiate into myofibroblasts during the progression of fibrosis. Therefore, inhibiting the differentiation of fibroblasts into myofibroblasts prevents further growth of myofibroblasts.

[0707] 4.6.5. Reversion of myofibroblasts to tissue epithelial cells

[0708] The above CSF3R inhibitor can induce or promote the reversion of myofibroblasts back to epithelial cells. As described above, just as epithelial cells can transform into myofibroblasts through epithelial-mesenchymal transition, the reverse process of epithelial-mesenchymal transition can occur, allowing myofibroblasts to revert to epithelial cells in normal tissues. This can work in conjunction with the reduction of myofibroblasts due to apoptosis and the inhibition of the dedifferentiation of epithelial cells into myofibroblasts. The results of Experimental Example 3 provide indirect evidence that myofibroblasts revert to tissue epithelial cells.

[0709] 4.6.6. Reversion of myofibroblasts to fibroblasts

[0710] The above CSF3R inhibitor can induce or promote the reversion of myofibroblasts back to fibroblasts. Just as fibroblasts can differentiate into myofibroblasts as described above, the reverse process of differentiation can occur, allowing myofibroblasts to revert back to fibroblasts. This can work in conjunction with the reduction of myofibroblasts due to apoptosis and the inhibition of fibroblast differentiation into myofibroblasts.

[0711] 4.6.7. Biomarkers associated with inhibition of epithelial-mesenchymal transition, or the reversion of myofibroblasts to tissue epithelial cells and / or fibroblasts.

[0712] Biomarkers associated with epithelial-mesenchymal transition (EMT), or the reversion of myofibroblasts to tissue epithelial cells, include E-cadherin, Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin. Specifically, when EMT occurs, the expression level of E-cadherin may decrease, and / or the expression level of one or more markers selected from Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin may increase. Conversely, when the expression level of E-cadherin increases, and / or the expression level of one or more markers selected from Snail1, Snail2, Zeb1, Zeb2, Vimentin, N-cadherin, and Fibronectin decreases, it can be interpreted that 1) EMT is inhibited and differentiation into myofibroblasts no longer occurs, or 2) myofibroblasts revert to tissue epithelial cells.

[0713] 4.7. Advantages of CSF3R inhibitors for fibrosis treatment 1 - Fibrosis treatment effect

[0714] When the anti-fibrosis treatment agent containing the CSF3R inhibitor is administered to a tissue where fibrosis has occurred, it has the effect of 1) decomposing accumulated myofibroblast-derived extracellular matrix, particularly collagen, and 2) reducing myofibroblasts in the tissue to restore it to normal tissue. This means that the anti-fibrosis treatment agent containing the CSF3R inhibitor has a direct and complete anti-fibrosis treatment effect. The inventors of the present disclosure have experimentally confirmed that the accumulated extracellular matrix in the tissue where fibrosis has occurred is decomposed, and the tissue where fibrosis has occurred returns to normal tissue. Compared to the fact that the conventional anti-fibrosis treatment agent only has an incomplete treatment effect on fibrosis, the anti-fibrosis treatment agent containing the CSF3R inhibitor has a complete treatment effect compared to the conventional anti-fibrosis treatment agent, and thus can be a superior treatment agent.

[0715] The reason why anti-fibrotic agents containing CSF3R inhibitors have a complete therapeutic effect compared to conventional anti-fibrotic agents may be because the degree to which the anti-fibrotic agents containing CSF3R inhibitors inhibit the TGF-β-mediated signaling pathway is superior to that of conventional anti-fibrotic agents. The reason why anti-fibrotic agents containing CSF3R inhibitors have a complete therapeutic effect compared to conventional anti-fibrotic agents may be because conventional anti-fibrotic agents only inhibit the TGF-β-mediated signaling pathway, whereas anti-fibrotic agents containing CSF3R inhibitors can inhibit not only the TGF-β-mediated signaling pathway but also non-TGF-β-mediated signaling pathways; the positive feedback loop of CSF3R; and the positive feedback loop of TGF-β.

[0716] 4.8. Advantage 2 of CSF3R inhibitor-based fibrosis treatment - Therapeutic effect regardless of the type of tissue where fibrosis occurs

[0717] The present inventors have revealed that CSF3R functions as a key regulator in the development of pulmonary fibrosis. Accordingly, pulmonary fibrosis treatments containing CSF3R inhibitors have a more fundamental mechanism of action than conventional pulmonary fibrosis treatments and exhibit true therapeutic effects. Furthermore, the present inventors experimentally demonstrated that CSF3R is also crucially involved in fibrosis in skin, liver, and kidney cells. These findings reasonably suggest that CSF3R is a key regulator (or upstream regulator) involved in tissue fibrosis, regardless of tissue type. Furthermore, this provides the basis for predicting that CSF3R inhibitors may have therapeutic effects on fibrosis regardless of tissue type.

[0718] Specifically, the inventors of the present disclosure have revealed the pathogenesis and / or treatment mechanism of fibrosis through Experimental Example 2, and have confirmed through Experimental Example 3 that a CSF3R inhibitor has a therapeutic effect on fibrosis occurring in various tissues. For example, Experimental Examples 3.1 and 3.2 confirmed that a CSF3R inhibitor has a therapeutic effect on pulmonary fibrosis. For example, Experimental Examples 3.3 and 3.4 confirmed that a CSF3R inhibitor has a therapeutic effect on skin fibrosis. For example, Experimental Example 3.5 confirmed that a CSF3R inhibitor has a therapeutic effect on liver fibrosis. For example, Experimental Example 3.6 confirmed that a CSF3R inhibitor has a therapeutic effect on renal fibrosis.

[0719] Therefore, the anti-fibrotic agent comprising the CSF3R inhibitor disclosed herein can act as an effective treatment regardless of the type of tissue in which fibrosis occurs.

[0720] In one specific example, the tissue in which the fibrosis occurs may be any one tissue selected from the group consisting of skin; brain; nervous system; heart; bone marrow; liver; gut; joint; eye; lung; kidney; retroperitoneum; mediastinum; and pancreas.

[0721] In one specific example, the CSF3R inhibitor has a therapeutic effect on skin fibrosis.

[0722] In one specific example, the CSF3R inhibitor has a therapeutic effect on cerebral fibrosis.

[0723] In one specific example, the CSF3R inhibitor has a therapeutic effect on neurofibromatosis.

[0724] In one specific example, the CSF3R inhibitor has a therapeutic effect on cardiac fibrosis.

[0725] In one specific example, the CSF3R inhibitor has a therapeutic effect on myelofibrosis.

[0726] In one specific example, the CSF3R inhibitor has a therapeutic effect on liver fibrosis.

[0727] In one specific example, the CSF3R inhibitor has a therapeutic effect on intestinal fibrosis.

[0728] In one specific example, the CSF3R inhibitor has a therapeutic effect on arthrofibrosis.

[0729] In one specific example, the CSF3R inhibitor has an anti-fibrotic effect.

[0730] In one specific example, the CSF3R inhibitor has a therapeutic effect on optic fibrosis.

[0731] In one specific example, the CSF3R inhibitor has a therapeutic effect on pulmonary fibrosis.

[0732] In one specific example, the CSF3R inhibitor has a therapeutic effect on renal fibrosis.

[0733] In one specific example, the CSF3R inhibitor has a therapeutic effect on retroperitoneal fibrosis.

[0734] In one specific example, the CSF3R inhibitor has a therapeutic effect on mediastinal fibrosis.

[0735] In one specific example, the CSF3R inhibitor has a therapeutic effect on pancreatic fibrosis.

[0736] 4.9. Pharmaceutical composition for treating fibrosis comprising a CSF3R inhibitor

[0737] 4.9.1. Overview of pharmaceutical compositions for treating fibrosis comprising CSF3R inhibitors

[0738] The present disclosure discloses a pharmaceutical composition for treating fibrosis comprising a CSF3R inhibitor. The pharmaceutical composition for treating fibrosis comprises a therapeutically effective amount of a CSF3R inhibitor. In one embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3R inhibitor and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3R inhibitor and an adjuvant. In another embodiment, the pharmaceutical composition for treating fibrosis may comprise a therapeutically effective amount of a CSF3R inhibitor; a pharmaceutically acceptable carrier; and an adjuvant. Here, the pharmaceutically acceptable carrier may be for appropriate formulation of the pharmaceutical composition for treating fibrosis.

[0739] 4.9.2. Target disease - Fibrosis

[0740] The pharmaceutical composition for treating fibrosis is intended to treat fibrosis, or a disease, condition, and / or symptom associated with fibrosis. The fibrosis or a disease, condition, and / or symptom associated with fibrosis includes all those recognizable to a person skilled in the art.

[0741] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0742] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0743] 4.9.3. Examples of formulations of pharmaceutical compositions for treating fibrosis

[0744] In one specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor may be formulated as a troche, lozenge, tablet, aqueous suspension, oily suspension, prepared powder, granules, emulsion, hard capsule, soft capsule, syrup or elixir. In another specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor may be formulated as an injection, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil or cream. In another specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor may be formulated as an injection. Specifically, a therapeutically effective amount of the CSF3R inhibitor may be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which may be formulated for unit dose in ampoules or vials. In another specific example, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor can be formulated as an aerosol after preparing a water-dispersed concentrate or wet powder by mixing a propellant and other additives. In another specific example, when the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor is formulated for transdermal use, an ointment, cream, powder for application, oil, external skin preparation, etc. can be prepared by adding animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as a carrier to a therapeutically effective amount of the CSF3R inhibitor.

[0745] 4.9.4. Examples of pharmaceutically acceptable carriers

[0746] Pharmaceutically acceptable carriers are those commonly used in formulations, and include, but are not limited to, saline solution, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc., and may further include other conventional additives such as antioxidants and buffers as needed. In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, each ingredient can be preferably formulated using the methods disclosed in Remington's Pharmaceutical Sciences (19th edition, 1995). In one embodiment, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor comprises a pharmaceutically acceptable carrier, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; an excipient such as dicalcium phosphate; a disintegrant such as corn starch or sweet potato starch; a lubricant such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; a sweetener; a fragrance; a syrup; a liquid carrier such as a fatty oil; a sterile aqueous solution; propylene glycol; polyethylene glycol; an injectable ester such as ethyl oleate; a suspension; an emulsion; a freeze-dried preparation; an external preparation; a stabilizer; a buffer; an animal oil; a vegetable oil; a wax; a paraffin; a starch; a tragacanth; a cellulose derivative; a polyethylene glycol; a silicone; a bentonite; a silica; a talc; zinc oxide, or a suitable combination thereof.

[0747] 4.10. Treatment of fibrosis using CSF3R inhibitors

[0748] 4.10.1. Overview of Fibrosis Treatment Using CSF3 Inhibitors

[0749] The present disclosure discloses a method for treating fibrosis using the CSF3R inhibitor. The method for treating fibrosis comprises administering a therapeutically effective amount of the CSF3R inhibitor to a subject using an appropriate administration method, an appropriate administration regimen, and an appropriate dosage. In one embodiment, the method for treating fibrosis comprises administering a therapeutically effective CSF3R inhibitor in an appropriate formulation, i.e., a pharmaceutical composition for treating fibrosis, to a subject using an appropriate administration method, an appropriate administration regimen, and an appropriate dosage. In one embodiment, the subject of the method for treating fibrosis may be a human or a non-human animal. In one embodiment, the subject of the method for treating fibrosis may be a subject suffering from fibrosis, or a disease, disorder, and / or symptom associated with fibrosis.

[0750] 4.10.2. Diseases to be treated - Fibrosis

[0751] The above fibrosis treatment method is intended to treat fibrosis, or a fibrosis-related disease, condition, and / or symptom. The fibrosis or fibrosis-related disease, condition, and / or symptom includes all those recognizable to a person skilled in the art.

[0752] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0753] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0754] 4.10.3. Examples of Dosage Methods

[0755] The above-described method for treating fibrosis may involve administering to a subject a therapeutically effective CSF3R inhibitor, appropriately formulated, via an appropriate administration method. In one embodiment, the above-described method for treating fibrosis may involve administering a therapeutically effective CSF3R inhibitor, appropriately formulated, orally or parenterally. The parenteral administration method may include intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, and subcutaneous administration.

[0756] Various criteria related to administration (dosage, cycle, frequency, etc.) can be adjusted by the physician according to the individual patient's needs based on clinical examination during the administration process.

[0757] 4.10.4. Dosage examples

[0758] The above-mentioned method for treating fibrosis may comprise administering to a subject an appropriate dosage of a therapeutically effective CSF3R inhibitor, appropriately formulated. In one embodiment, the dosage may be about 0.01 mg to 1000 mg per kg of the subject's body weight, based on the CSF3R inhibitor.

[0759] 4.10.5. Example of Dosage Cycle

[0760] The above method for treating fibrosis may be to administer to a subject an appropriately formulated therapeutically effective CSF3R inhibitor at an appropriate dosing cycle. In one embodiment, the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor at the appropriate dosage may be administered once a day. In another example, the dosing cycle may be to administer the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor at the appropriate dosage twice or more a day in divided doses. In another example, the dosing cycle may be to administer the pharmaceutical composition for treating fibrosis comprising the CSF3R inhibitor at the appropriate dosage at intervals of 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 1 week, 2 weeks, 1 month, and / or 3 months.

[0761] 4.11. Therapeutic Uses of CSF3R Inhibitors

[0762] The present disclosure discloses the use of a CSF3R inhibitor for treating fibrosis. In one embodiment, the present disclosure discloses a CSF3R inhibitor for treating fibrosis, or a fibrosis-associated disease, condition, and / or symptom.

[0763] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0764] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0765] 4.12. Use of CSF3R inhibitors for therapeutic purposes

[0766] The present disclosure discloses the use of a CSF3R inhibitor for the manufacture of a fibrosis treatment agent. In one specific example, the present disclosure discloses the use of a CSF3R inhibitor for the manufacture of a medicament for the treatment of fibrosis, or a fibrosis-related disease, condition, and / or symptom.

[0767] In one specific example, the fibrosis or fibrosis-associated disease, condition, and / or symptom may be one described in the <3.14. Fibrosis Disease Examples> table of contents.

[0768] As a specific example, the tissue in which the fibrosis occurs may be described in the table of contents of <3.13. Examples of tissues in which fibrosis occurs>.

[0769] 4.13. Examples of tissues where fibrosis occurs

[0770] Fibrosis can develop in virtually any tissue in the body, and it is known that many tissues within the body can become fibrotic, leading to a variety of diseases, conditions, and / or conditions. For example, fibrosis has been reported to occur in tissues such as the skin, brain, nervous system, heart, bone marrow, liver, gut, joints, eyes, lungs, kidneys, retroperitoneum, mediastinum, and / or pancreas.

[0771] 4.14. Examples of Fibrotic Diseases

[0772] The fibrosis or fibrosis-associated diseases, conditions, and / or symptoms described herein encompass all those recognizable to those of ordinary skill in the art. In one specific example, the fibrosis or fibrosis-associated diseases, conditions, and / or symptoms may be those described in the <3.14. Fibrosis Diseases Examples> table of contents.

[0773] 4.15. Examples of CSF3R inhibitors

[0774] The CSF3R inhibitor described in the present disclosure is not limited to anything that can inhibit the function of CSF3R. As described above, the CSF3R inhibitor may 1) interfere with the interaction of the CSF3R protein with other molecules, or 2) inhibit the expression of the CSF3R protein, thereby reducing the absolute amount of the CSF3R protein that interacts with other molecules, thereby resulting in the interference with the function of the CSF3R protein. Examples of CSF3R inhibitors that can have such an effect include, but are not limited to, a nucleic acid that silences the mRNA of the CSF3R gene, a CRISPR / Cas system that knocks out or knocks down the CSF3R gene, a compound that inhibits the CSF3R protein, or an aptamer that inhibits the CSF3R protein.

[0775] As a specific example, the CSF3R inhibitor may be one described in the table of contents <2.9. Examples of CSF3R inhibitors> and its sub-tables <2.9.1. Nucleic acid that silences mRNA of CSF3R gene> to <2.9.6. Substance 3 that inhibits CSF3R protein - Antibody>.

[0776]

[0777] 5. Anti-CSF3 antibodies

[0778] 5.1. Anti-CSF3 Antibodies - Overview

[0779] In one aspect of the present disclosure, an antibody against colony-stimulating factor 3 (CSF3) is disclosed. In the present disclosure, the anti-CSF3 antibody may be referred to as an anti-CSF3 antibody.

[0780] The anti-CSF3 antibody may bind to CSF3 or a fragment of CSF3. In one embodiment, the anti-CSF3 antibody may specifically bind to CSF3 or a fragment of CSF3. In another embodiment, the anti-CSF3 antibody may inhibit or interfere with the binding of CSF3 to CSF3R. In another embodiment, the anti-CSF3 antibody may inhibit the function of CSF3. In another embodiment, the anti-CSF3 antibody may be an anti-CSF3 neutralizing antibody. In another embodiment, the anti-CSF3 antibody may be a monoclonal antibody that specifically binds to human CSF3 protein.

[0781] Table 1 below lists amino acid sequences that may be included in CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, CDRL3, FRH1, FRH2, FRH3, FRH4, FRL1, FRL2, FRL3, or FRL4 of the anti-CSF3 antibody.

[0782] Table 2 below lists amino acid sequences that may be included in the VH and VL of the anti-CSF3 antibody.

[0783] [Table 1]

[0784]

[0785] [Table 2]

[0786]

[0787]

[0788]

[0789] 5.2. Sequence of the variable region of anti-CSF3 antibodies

[0790] Sequence of CDRH1

[0791] In one specific example, the amino acid sequence of CDRH1 of the anti-CSF3 antibody may be SEQ ID NO: 1 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 1.

[0792] In one specific example, the amino acid sequence of CDRH1 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 1.

[0793] As a specific example, the amino acid sequence of CDRH1 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 1.

[0794] In one specific example, the VH of the anti-CSF3 antibody may comprise a CDRH1 comprising an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 1.

[0795] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising an amino acid sequence identical to SEQ ID NO: 1.

[0796] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH1 represented by the amino acid sequence of SEQ ID NO: 1.

[0797] Sequence of CDRH2

[0798] In one specific example, the amino acid sequence of CDRH2 of the anti-CSF3 antibody may be SEQ ID NO: 2 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 2.

[0799] In one specific example, the amino acid sequence of CDRH2 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 2.

[0800] As a specific example, the amino acid sequence of CDRH2 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 2.

[0801] In one specific example, the VH of the anti-CSF3 antibody may comprise a CDRH2 comprising an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 2.

[0802] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH2 comprising an amino acid sequence identical to SEQ ID NO: 2.

[0803] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH2 represented by the amino acid sequence of SEQ ID NO: 2.

[0804] Sequence of CDRH3

[0805] In one specific example, the amino acid sequence of CDRH3 of the anti-CSF3 antibody may be an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of the amino acid sequences of SEQ ID NOs: 3 to 20.

[0806] In one specific example, the amino acid sequence of CDRH3 of the anti-CSF3 antibody may include any one of the amino acid sequences of SEQ ID NOs: 3 to 20.

[0807] In one specific example, the amino acid sequence of CDRH3 of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 3 to 20.

[0808] In one specific example, the VH of the anti-CSF3 antibody may comprise a CDRH3 comprising an amino acid sequence of any one of SEQ ID NOs: 3 to 20 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 3 to 20.

[0809] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH3 comprising an amino acid sequence of any one of SEQ ID NOs: 3 to 20.

[0810] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH3 represented by any one of the amino acid sequences of SEQ ID NOs: 3 to 20.

[0811] Sequence of CDRL1

[0812] In one specific example, the amino acid sequence of CDRL1 of the anti-CSF3 antibody may be an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of the amino acid sequences of SEQ ID NOs: 21 to 24.

[0813] In one specific example, the amino acid sequence of CDRL1 of the anti-CSF3 antibody may include any one of the amino acid sequences of SEQ ID NOs: 21 to 24.

[0814] In one specific example, the amino acid sequence of CDRL1 of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 21 to 24.

[0815] In one specific example, the VL of the anti-CSF3 antibody may comprise a CDRL1 comprising an amino acid sequence of any one of SEQ ID NOs: 21 to 24 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 21 to 24.

[0816] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising an amino acid sequence of any one of SEQ ID NOs: 21 to 24.

[0817] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL1 represented by any one of the amino acid sequences of SEQ ID NOs: 21 to 24.

[0818] Sequence of CDRL2

[0819] In one specific example, the amino acid sequence of CDRL2 of the anti-CSF3 antibody may be YAS (SEQ ID NO: 25) or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to YAS (SEQ ID NO: 25).

[0820] As a specific example, the amino acid sequence of CDRL2 of the anti-CSF3 antibody may include the amino acid sequence of YAS (SEQ ID NO: 25).

[0821] As a specific example, the amino acid sequence of CDRL2 of the anti-CSF3 antibody may be the amino acid sequence of YAS (SEQ ID NO: 25).

[0822] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL2 comprising an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to the sequence of YAS (SEQ ID NO: 25).

[0823] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL2 comprising the amino acid sequence of YAS (SEQ ID NO: 25).

[0824] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL2 represented by the amino acid sequence of YAS (SEQ ID NO: 25).

[0825] Sequence of CDRL3

[0826] In one specific example, the amino acid sequence of CDRL3 of the anti-CSF3 antibody may be SEQ ID NO: 26 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 26.

[0827] As a specific example, the amino acid sequence of CDRL3 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 26.

[0828] As a specific example, the amino acid sequence of CDRL3 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 26.

[0829] In one specific example, the VL of the anti-CSF3 antibody may comprise a CDRL3 comprising an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 26 or a sequence of SEQ ID NO: 26.

[0830] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL3 comprising the amino acid sequence of SEQ ID NO: 26.

[0831] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL3 represented by the amino acid sequence of SEQ ID NO: 26.

[0832] CDRH1, CDRH2, and CDRH3

[0833] In one specific example, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising (or being represented by) the amino acid sequence of SEQ ID NO: 1, CDRH2 comprising (or being represented by) the amino acid sequence of SEQ ID NO: 2, and CDRH3 comprising (or being represented by) any one of the amino acid sequences of SEQ ID NOs: 3 to 20.

[0834] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 3 (or represented by the amino acid sequence of SEQ ID NO: 3).

[0835] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 4 (or represented by the amino acid sequence of SEQ ID NO: 4).

[0836] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 5 (or represented by the amino acid sequence of SEQ ID NO: 5).

[0837] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 6 (or represented by the amino acid sequence of SEQ ID NO: 6).

[0838] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 7 (or represented by the amino acid sequence of SEQ ID NO: 7).

[0839] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 8 (or represented by the amino acid sequence of SEQ ID NO: 8).

[0840] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 9 (or represented by the amino acid sequence of SEQ ID NO: 9).

[0841] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 10 (or represented by the amino acid sequence of SEQ ID NO: 10).

[0842] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 11 (or represented by the amino acid sequence of SEQ ID NO: 11).

[0843] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 12 (or represented by the amino acid sequence of SEQ ID NO: 12).

[0844] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 13 (or represented by the amino acid sequence of SEQ ID NO: 13).

[0845] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 14 (or represented by the amino acid sequence of SEQ ID NO: 14).

[0846] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 15 (or represented by the amino acid sequence of SEQ ID NO: 15).

[0847] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 16 (or represented by the amino acid sequence of SEQ ID NO: 16).

[0848] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 17 (or represented by the amino acid sequence of SEQ ID NO: 17).

[0849] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 18 (or represented by the amino acid sequence of SEQ ID NO: 18).

[0850] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising the amino acid sequence of SEQ ID NO: 1 (or represented by the amino acid sequence of SEQ ID NO: 1), CDRH2 comprising the amino acid sequence of SEQ ID NO: 2 (or represented by the amino acid sequence of SEQ ID NO: 2), and CDRH3 comprising the amino acid sequence of SEQ ID NO: 19 (or represented by the amino acid sequence of SEQ ID NO: 19).

[0851] Preferably, the VH of the anti-CSF3 antibody may comprise CDRH1 comprising (or represented by) the amino acid sequence of SEQ ID NO: 1, CDRH2 comprising (or represented by) the amino acid sequence of SEQ ID NO: 2, and CDRH3 comprising (or represented by) the amino acid sequence of SEQ ID NO: 20.

[0852] CDRL1, CDRL2, and CDRL3

[0853] In one specific example, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising (or being represented by) the amino acid sequence of any one of SEQ ID NOs: 21 to 24, CDRL2 comprising (or being represented by the amino acid sequence of YAS (SEQ ID NO: 25)), and CDRL3 comprising (or being represented by the amino acid sequence of SEQ ID NO: 26).

[0854] Preferably, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising the amino acid sequence of SEQ ID NO: 21 (or represented by the amino acid sequence of SEQ ID NO: 21), CDRL2 comprising the amino acid sequence of YAS (SEQ ID NO: 25) (or represented by the amino acid sequence of YAS (SEQ ID NO: 25)), and CDRL3 comprising the amino acid sequence of SEQ ID NO: 26 (or represented by the amino acid sequence of SEQ ID NO: 26).

[0855] Preferably, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising the amino acid sequence of SEQ ID NO: 22 (or represented by the amino acid sequence of SEQ ID NO: 22), CDRL2 comprising the amino acid sequence of YAS (SEQ ID NO: 25) (or represented by the amino acid sequence of YAS (SEQ ID NO: 25)), and CDRL3 comprising the amino acid sequence of SEQ ID NO: 26 (or represented by the amino acid sequence of SEQ ID NO: 26).

[0856] Preferably, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising the amino acid sequence of SEQ ID NO: 23 (or represented by the amino acid sequence of SEQ ID NO: 23), CDRL2 comprising the amino acid sequence of YAS (SEQ ID NO: 25) (or represented by the amino acid sequence of YAS (SEQ ID NO: 25)), and CDRL3 comprising the amino acid sequence of SEQ ID NO: 26 (or represented by the amino acid sequence of SEQ ID NO: 26).

[0857] Preferably, the VL of the anti-CSF3 antibody may comprise CDRL1 comprising the amino acid sequence of SEQ ID NO: 24 (or represented by the amino acid sequence of SEQ ID NO: 24), CDRL2 comprising the amino acid sequence of YAS (SEQ ID NO: 25) (or represented by the amino acid sequence of YAS (SEQ ID NO: 25)), and CDRL3 comprising the amino acid sequence of SEQ ID NO: 26 (or represented by the amino acid sequence of SEQ ID NO: 26).

[0858] CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3

[0859] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0860] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0861] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0862] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 22, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0863] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0864] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 8, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0865] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 9, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0866] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 10, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0867] As a specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 11, SEQ ID NO: 23, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0868] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 12, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0869] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 13, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0870] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 14, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0871] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 15, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0872] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 16, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0873] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 17, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0874] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 18, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0875] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 24, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0876] In one specific example, the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 20, SEQ ID NO: 21, YAS (SEQ ID NO: 25), and SEQ ID NO: 26, respectively.

[0877] Sequence of FRH1

[0878] In one specific example, the amino acid sequence of FRH1 of the anti-CSF3 antibody may be an amino acid sequence of any one of SEQ ID NOs: 27 to 28 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 27 to 28.

[0879] In one specific example, the amino acid sequence of FRH1 of the anti-CSF3 antibody may include any one of the amino acid sequences of SEQ ID NOs: 27 to 28.

[0880] In one specific example, the amino acid sequence of FRH1 of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 27 to 28.

[0881] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH1 comprising an amino acid sequence of any one of SEQ ID NOs: 27 to 28 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 27 to 28.

[0882] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH1 comprising an amino acid sequence identical to any one of SEQ ID NOs: 27 to 28.

[0883] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH1 represented by any one of the amino acid sequences of SEQ ID NOs: 27 to 28.

[0884] Sequence of FRH2

[0885] In one specific example, the amino acid sequence of FRH2 of the anti-CSF3 antibody may be SEQ ID NO: 29 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 29.

[0886] As a specific example, the amino acid sequence of FRH2 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 29.

[0887] As a specific example, the amino acid sequence of FRH2 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 29.

[0888] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH2 comprising SEQ ID NO: 29 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 29.

[0889] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH2 comprising an amino acid sequence identical to SEQ ID NO: 29.

[0890] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH2 represented by the amino acid sequence of SEQ ID NO: 29.

[0891] Sequence of FRH3

[0892] In one specific example, the amino acid sequence of FRH3 of the anti-CSF3 antibody may be an amino acid sequence of any one of SEQ ID NOs: 30 to 31 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 30 to 31.

[0893] In one specific example, the amino acid sequence of FRH3 of the anti-CSF3 antibody may include any one of the amino acid sequences of SEQ ID NOs: 30 to 31.

[0894] In one specific example, the amino acid sequence of FRH3 of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 30 to 31.

[0895] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH3 comprising an amino acid sequence of any one of SEQ ID NOs: 30 to 31 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 30 to 31.

[0896] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH3 comprising an amino acid sequence identical to any one of SEQ ID NOs: 30 to 31.

[0897] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH3 represented by any one of the amino acid sequences of SEQ ID NOs: 30 to 31.

[0898] Sequence of FRH4

[0899] In one specific example, the amino acid sequence of FRH4 of the anti-CSF3 antibody may be SEQ ID NO: 32 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 32.

[0900] As a specific example, the amino acid sequence of FRH4 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 32.

[0901] As a specific example, the amino acid sequence of FRH4 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 32.

[0902] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH4 comprising SEQ ID NO: 32 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 32.

[0903] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH4 comprising an amino acid sequence identical to SEQ ID NO: 32.

[0904] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH4 represented by the amino acid sequence of SEQ ID NO: 32.

[0905] Sequence of FRL1

[0906] In one specific example, the amino acid sequence of FRL1 of the anti-CSF3 antibody may be SEQ ID NO: 33 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 33.

[0907] As a specific example, the amino acid sequence of FRL1 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 33.

[0908] As a specific example, the amino acid sequence of FRL1 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 33.

[0909] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL1 comprising SEQ ID NO: 33 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 33.

[0910] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL1 comprising an amino acid sequence identical to SEQ ID NO: 33.

[0911] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL1 represented by the amino acid sequence of SEQ ID NO: 33.

[0912] Sequence of FRL2

[0913] In one specific example, the amino acid sequence of FRL2 of the anti-CSF3 antibody may be SEQ ID NO: 34 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 34.

[0914] As a specific example, the amino acid sequence of FRL2 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 34.

[0915] As a specific example, the amino acid sequence of FRL2 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 34.

[0916] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL2 comprising SEQ ID NO: 34 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 34.

[0917] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL2 comprising an amino acid sequence identical to SEQ ID NO: 34.

[0918] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL2 represented by the amino acid sequence of SEQ ID NO: 34.

[0919] Sequence of FRL3

[0920] In one specific example, the amino acid sequence of FRL3 of the anti-CSF3 antibody may be SEQ ID NO: 35 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 35.

[0921] As a specific example, the amino acid sequence of FRL3 of the anti-CSF3 antibody may include the amino acid sequence of SEQ ID NO: 35.

[0922] As a specific example, the amino acid sequence of FRL3 of the anti-CSF3 antibody may be the amino acid sequence of SEQ ID NO: 35.

[0923] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL3 comprising SEQ ID NO: 35 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 35.

[0924] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL3 comprising an amino acid sequence identical to SEQ ID NO: 35.

[0925] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL3 represented by the amino acid sequence of SEQ ID NO: 35.

[0926] Sequence of FRL4

[0927] In one specific example, the amino acid sequence of FRL4 of the anti-CSF3 antibody may be an amino acid sequence of any one of SEQ ID NOs: 36 to 37 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 36 to 37.

[0928] In one specific example, the amino acid sequence of FRL4 of the anti-CSF3 antibody may include any one of the amino acid sequences of SEQ ID NOs: 36 to 37.

[0929] In one specific example, the amino acid sequence of FRL4 of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 36 to 37.

[0930] In one specific example, the VL of the anti-CSF3 antibody may comprise an FRL4 comprising an amino acid sequence of any one of SEQ ID NOs: 36 to 37 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 36 to 37.

[0931] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL4 comprising an amino acid sequence identical to any one of SEQ ID NOs: 36 to 37.

[0932] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL4 represented by any one of the amino acid sequences of SEQ ID NOs: 36 to 37.

[0933] FRH1, FRH2, FRH3, and FRH4

[0934] In one specific example, the VH of the anti-CSF3 antibody may comprise FRH1 comprising (or being represented by) the amino acid sequence of any one of SEQ ID NOs: 27 to 28, FRH2 comprising (or being represented by the amino acid sequence of SEQ ID NO: 29), FRH3 comprising (or being represented by the amino acid sequence of SEQ ID NO: 30 to 31), and FRH4 comprising (or being represented by the amino acid sequence of SEQ ID NO: 32).

[0935] Preferably, the VH of the anti-CSF3 antibody may comprise FRH1 comprising the amino acid sequence of SEQ ID NO: 27 (or represented by the amino acid sequence of SEQ ID NO: 27), FRH2 comprising the amino acid sequence of SEQ ID NO: 29 (or represented by the amino acid sequence of SEQ ID NO: 29), FRH3 comprising the amino acid sequence of SEQ ID NO: 30 (or represented by the amino acid sequence of SEQ ID NO: 30), and FRH4 comprising the amino acid sequence of SEQ ID NO: 32 (or represented by the amino acid sequence of SEQ ID NO: 32).

[0936] Preferably, the VH of the anti-CSF3 antibody may comprise FRH1 comprising the amino acid sequence of SEQ ID NO: 28 (or represented by the amino acid sequence of SEQ ID NO: 28), FRH2 comprising the amino acid sequence of SEQ ID NO: 29 (or represented by the amino acid sequence of SEQ ID NO: 29), FRH3 comprising the amino acid sequence of SEQ ID NO: 31 (or represented by the amino acid sequence of SEQ ID NO: 31), and FRH4 comprising the amino acid sequence of SEQ ID NO: 32 (or represented by the amino acid sequence of SEQ ID NO: 32).

[0937] FRL1, FRL2, FRL3, and FRL4

[0938] In one specific example, the VL of the anti-CSF3 antibody may comprise FRL1 comprising (or being represented by) the amino acid sequence of SEQ ID NO: 33, FRL2 comprising (or being represented by) the amino acid sequence of SEQ ID NO: 34, FRL3 comprising (or being represented by) the amino acid sequence of SEQ ID NO: 35, and FRL4 comprising (or being represented by) any one of the amino acid sequences of SEQ ID NOs: 36 to 37.

[0939] Preferably, the VL of the anti-CSF3 antibody may comprise FRL1 comprising (or represented by) the amino acid sequence of SEQ ID NO: 33, FRL2 comprising (or represented by) the amino acid sequence of SEQ ID NO: 34, FRL3 comprising (or represented by) the amino acid sequence of SEQ ID NO: 35, and FRL4 comprising (or represented by) the amino acid sequence of SEQ ID NO: 36.

[0940] Preferably, the VL of the anti-CSF3 antibody may comprise FRL1 comprising (or represented by) the amino acid sequence of SEQ ID NO: 33, FRL2 comprising (or represented by) the amino acid sequence of SEQ ID NO: 34, FRL3 comprising (or represented by) the amino acid sequence of SEQ ID NO: 35, and FRL4 comprising (or represented by) the amino acid sequence of SEQ ID NO: 37.

[0941] FRH1, FRH2, FRH3, FRH4, FRL1, FRL2, FRL3, and FRL4

[0942] In one specific example, the amino acid sequences of FRH1, FRH2, FRH3, FRH4, FRL1, FRL2, FRL3, and FRL4 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively.

[0943] In one specific example, the amino acid sequences of FRH1, FRH2, FRH3, FRH4, FRL1, FRL2, FRL3, and FRL4 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 37, respectively.

[0944] In one specific example, the amino acid sequences of FRH1, FRH2, FRH3, FRH4, FRL1, FRL2, FRL3, and FRL4 of the variable region of the anti-CSF3 antibody may be the amino acid sequences of SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively.

[0945] VH's sequence

[0946] In one specific example, the VH of the anti-CSF3 antibody may comprise an amino acid sequence of any one of SEQ ID NOs: 38 to 55 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 38 to 55.

[0947] In one specific example, the VH of the anti-CSF3 antibody may comprise any one of the amino acid sequences of SEQ ID NOs: 38 to 55.

[0948] In one specific example, the amino acid sequence of the VH of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 38 to 55.

[0949] VL's sequence

[0950] In one specific example, the VL of the anti-CSF3 antibody may comprise an amino acid sequence of any one of SEQ ID NOs: 56 to 60, or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 56 to 60.

[0951] In one specific example, the VL of the anti-CSF3 antibody may comprise any one of the amino acid sequences of SEQ ID NOs: 56 to 60.

[0952] In one specific example, the amino acid sequence of the VL of the anti-CSF3 antibody may be any one of the amino acid sequences of SEQ ID NOs: 56 to 60.

[0953] VH and VL

[0954] In one specific example, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) an amino acid sequence of any one of SEQ ID NOs: 38 to 55, and a VL comprising (or being represented by an amino acid sequence of any one of SEQ ID NOs: 56 to 60).

[0955] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 39, and a VL comprising (or being represented by the amino acid sequence of SEQ ID NO: 56).

[0956] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 40, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 57.

[0957] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 41, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 58.

[0958] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 42, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0959] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 43, and a VL comprising (or being represented by the amino acid sequence of SEQ ID NO: 56).

[0960] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 44, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0961] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 45, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0962] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 46, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 59.

[0963] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 47, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0964] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 48, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0965] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 49, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0966] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 50, and a VL comprising (or being represented by the amino acid sequence of SEQ ID NO: 56).

[0967] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 51, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0968] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 52, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0969] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 53, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0970] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 54, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 60.

[0971] Preferably, the variable region of the anti-CSF3 antibody may comprise a VH comprising (or being represented by) the amino acid sequence of SEQ ID NO: 55, and a VL comprising (or being represented by) the amino acid sequence of SEQ ID NO: 56.

[0972]

[0973]

[0974] In the table of contents below <<5.3. Sequence of constant region of anti-CSF3 antibody>> and <<5.4. Sequence of heavy or light chain of anti-CSF3 antibody>>, the description is made assuming that the type of anti-CSF3 antibody of the present disclosure is IgG1.

[0975] Table 3 below lists amino acid sequences that may be included in the CH, CL, heavy chain region, and light chain region of the anti-CSF3 antibody when the type of anti-CSF3 antibody of the present disclosure is IgG1.

[0976] [Table 3]

[0977]

[0978]

[0979]

[0980]

[0981]

[0982]

[0983] 5.3. Sequence of the constant region of anti-CSF3 antibodies

[0984] Sequence of the heavy chain constant region (CH)

[0985] In one specific example, the CH of the anti-CSF3 antibody may comprise SEQ ID NO: 61 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 61.

[0986] In one specific example, the CH of the anti-CSF3 antibody may comprise the amino acid sequence of SEQ ID NO: 61.

[0987] As a specific example, the CH of the anti-CSF3 antibody may be represented by the amino acid sequence of SEQ ID NO: 61.

[0988] Sequence of the light chain constant region (CL)

[0989] In one specific example, the CL of the anti-CSF3 antibody may comprise SEQ ID NO: 62 or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to SEQ ID NO: 62.

[0990] In another specific example, the CL of the anti-CSF3 antibody may comprise the amino acid sequence of SEQ ID NO: 62.

[0991] As another specific example, the CL of the anti-CSF3 antibody may be represented by the amino acid sequence of SEQ ID NO: 62.

[0992]

[0993] 5.4. Heavy or light chain sequence of anti-CSF3 antibody

[0994] Heavy chain region sequence

[0995] In one specific example, the heavy chain region of the anti-CSF3 antibody may comprise an amino acid sequence of any one of SEQ ID NOs: 63 to 80, or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 63 to 80.

[0996] In another specific example, the heavy chain region of the anti-CSF3 antibody may comprise any one of the amino acid sequences of SEQ ID NOs: 63 to 80.

[0997] In another specific example, the heavy chain region of the anti-CSF3 antibody may be represented by any one of the amino acid sequences of SEQ ID NOs: 63 to 80.

[0998] Sequence of the light chain region

[0999] In one specific example, the light chain region of the anti-CSF3 antibody may comprise an amino acid sequence of any one of SEQ ID NOs: 81 to 85, or an amino acid sequence having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence similarity to any one of SEQ ID NOs: 81 to 85.

[1000] In another specific example, the light chain region of the anti-CSF3 antibody may comprise an amino acid sequence of any one of SEQ ID NOs: 81 to 85.

[1001] In another specific example, the light chain region of the anti-CSF3 antibody may be represented by any one of the amino acid sequences of SEQ ID NOs: 81 to 85.

[1002] Sequences of heavy and light chain regions

[1003] In one specific example, the anti-CSF3 antibody may comprise a heavy chain region comprising an amino acid sequence of any one of SEQ ID NOs: 63 to 80 (or represented by an amino acid sequence of any one of SEQ ID NOs: 63 to 80), and a light chain region comprising an amino acid sequence of any one of SEQ ID NOs: 81 to 85 (or represented by an amino acid sequence of any one of SEQ ID NOs: 81 to 85).

[1004] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or being represented by) the amino acid sequence of SEQ ID NO: 63, and a light chain region comprising (or being represented by the amino acid sequence of SEQ ID NO: 81).

[1005] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or being represented by) the amino acid sequence of SEQ ID NO: 64, and a light chain region comprising (or being represented by the amino acid sequence of SEQ ID NO: 81).

[1006] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or being represented by) the amino acid sequence of SEQ ID NO: 65, and a light chain region comprising (or being represented by the amino acid sequence of SEQ ID NO: 82).

[1007] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or being represented by) the amino acid sequence of SEQ ID NO: 66, and a light chain region comprising (or being represented by the amino acid sequence of SEQ ID NO: 83).

[1008] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 67, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1009] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 68, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1010] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 69, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1011] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 70, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1012] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 71, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 84).

[1013] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising the amino acid sequence of SEQ ID NO: 72 (or represented by the amino acid sequence of SEQ ID NO: 72), and a light chain region comprising the amino acid sequence of SEQ ID NO: 81 (or represented by the amino acid sequence of SEQ ID NO: 81).

[1014] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 73, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1015] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 74, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1016] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 75, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1017] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 76, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1018] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 77, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1019] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 78, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 81).

[1020] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or represented by) the amino acid sequence of SEQ ID NO: 79, and a light chain region comprising (or represented by the amino acid sequence of SEQ ID NO: 85).

[1021] Preferably, the anti-CSF3 antibody may comprise a heavy chain region comprising (or being represented by) the amino acid sequence of SEQ ID NO: 80, and a light chain region comprising (or being represented by the amino acid sequence of SEQ ID NO: 81).

[1022] 5.5. Composition according to the type of anti-CSF3 antibody

[1023] The type of the anti-CSF3 antibody of the present disclosure may be any one of IgG, IgA, IgE, IgM, and IgD. Depending on the type of the anti-CSF3 antibody, the structure of the antibody may be any one of a monomer, a dimer, a trimer, and a pentamer. That is, the number of heavy chains and light chains of the anti-CSF3 antibody may be any one of two heavy chains and two light chains, four heavy chains and four light chains, six heavy chains and six light chains, and ten heavy chains and ten light chains. At this time, two or more heavy chains included in the anti-CSF3 antibody may be the same heavy chain, or may be different heavy chains. For example, when the anti-CSF3 antibody includes two heavy chains, 1) both heavy chains may be the same, or 2) the two heavy chains may be different. For another example, if the anti-CSF3 antibody comprises four heavy chains, 1) all four heavy chains may be identical, 2) only three heavy chains may be identical, 4) only two heavy chains may be identical, or 4) all four heavy chains may be different from each other.

[1024] 5.6. Origin of anti-CSF3 antibodies

[1025] If it is of human origin

[1026] In one specific example, the anti-CSF3 antibody may be a human antibody. Specifically, the anti-CSF3 antibody may be a human antibody. In this case, the anti-CSF3 antibody being a human antibody means that the anti-CSF3 antibody is an antibody derived from human immunoglobulin. That is, it means that the entire amino acid sequence constituting the anti-CSF3 antibody (all types of CDR, FR, CH, CL, and hinge region, etc.) is composed of an amino acid sequence derived from human immunoglobulin.

[1027] If it is of non-human animal origin

[1028] In one specific example, the anti-CSF3 antibody may be an antibody derived from a non-human animal. In this case, the anti-CSF3 antibody being an antibody derived from a non-human animal means that the anti-CSF3 antibody is an antibody derived from an immunoglobulin of the non-human animal. That is, it means that all amino acid sequences constituting the anti-CSF3 antibody (all types of CDRs, FRs, CHs, CLs, hinge regions, etc.) are composed of amino acid sequences derived from an immunoglobulin of the non-human animal. In one specific example, the non-human animal from which the anti-CSF3 antibody is derived may be a mammal (e.g., a mouse, a rabbit, a cow, a goat, or a sheep, etc.).

[1029] In the case of non-human animals and human origin

[1030] In one specific example, the anti-CSF3 antibody may be an antibody derived from a human and / or non-human animal.

[1031] In one embodiment, the anti-CSF3 antibody may be a chimeric antibody. Specifically, the variable region of the anti-CSF3 antibody may be derived from an immunoglobulin of a non-human animal, and the constant region may be derived from a human immunoglobulin.

[1032] In one embodiment, the anti-CSF3 antibody may be a humanized antibody. Specifically, the constant and variable region FRs of the anti-CSF3 antibody may be derived from a human immunoglobulin, and the variable region CDRs may be derived from a non-human animal immunoglobulin. In this case, some residues of the variable region FRs may be derived from the non-human animal immunoglobulin.

[1033] 5.7. Fragments of anti-CSF3 antibodies

[1034] 5.7.1. Overview

[1035] In one aspect of the present disclosure, a fragment of an anti-CSF3 antibody is disclosed. The fragment of the anti-CSF3 antibody comprises a portion of the anti-CSF3 antibody of the present disclosure, and the fragment may have any type of form capable of binding to CSF3. Preferably, the fragment of the anti-CSF3 antibody may comprise an antigen-binding portion of the anti-CSF3 antibody. The fragment of the anti-CSF3 antibody comprises a portion of any one of the anti-CSF3 antibodies described in the table of contents of <<5.1. Anti-CSF3 Antibodies - Overview>> to <<5.6. Origin of Anti-CSF3 Antibodies>> above.

[1036] Below, examples of structures that fragments of the anti-CSF3 antibody may have are described, but are not limited thereto.

[1037] 5.7.2. Example of the structure of a fragment of an anti-CSF3 antibody

[1038] In one specific example, the fragment of the anti-CSF3 antibody may be selected from among F(ab), F(ab'), F(ab')2, monospecific F(ab')2, bispecific F(ab')2, single-chain variable fragment (scFv), scFv-Fc, and sdAb.

[1039] The above F(ab) means that it has a structure including the VH, CH1, VL, and CL of an antibody, and has only one antigen binding site.

[1040] The above F(ab') has a structure including the VH, CH1, VL, and CL of an antibody, and means that compared to Fab, it additionally has a hinge including one or more cysteine ​​residues at the C-terminal of CH1.

[1041] The above F(ab')2 means that two F(ab') are connected. At this time, the connection of the two F(ab') is due to a disulfide bond of the cysteine ​​residues in the hinge region, and the two connected F(ab') may be the same or different. The above monospecific F(ab')2 means that the antigen-binding portions of the two connected F(ab') of the F(ab')2 can bind to the same antigen. The above bispecific F(ab')2 means that the antigen-binding portions of the two connected F(ab') of the F(ab')2 can bind to different antigens.

[1042] The single-chain variable fragment (scFv) above means that it includes only VH and VL, and does not include the immunoglobulin domains (CL, CH1, CH2, CH3, and CH4) belonging to the constant region of an antibody. At this time, the VH and VL included in the scFv are generally covalently bonded to each other via a peptide linker. Specifically, the peptide linker connects the C-terminal of VH and the N-terminal of VL, or connects the N-terminal of VH and the C-terminal of VL. When two or three scFvs are connected, there may be types such as tandem di-scFv, diabody, tandem tri-scFv, and tribody. The scFv-Fc above means that it includes scFv, Ch2, and CH3. Specifically, the scFv-Fc means that scFv, Ch2, and CH3 are connected in that order.

[1043] The above single-domain antibody (sdAB) means one that contains only one of the variable regions of the antibody, VH or VL.

[1044] 5.7.3. Artificially engineered proteins containing fragments of anti-CSF3 antibodies

[1045] In one aspect of the present disclosure, an artificially engineered protein comprising a fragment of the anti-CSF3 antibody is disclosed. The artificially engineered protein comprises a portion of the anti-CSF3 antibody of the present disclosure, and the artificially engineered protein may have any type of conformation capable of binding to CSF3. Preferably, the artificially engineered protein may comprise an antigen-binding portion of the anti-CSF3 antibody. Alternatively, the artificially engineered protein may comprise a CDR region of the anti-CSF3 antibody. The artificially engineered protein comprises a portion of any one of the anti-CSF3 antibodies described in the table of contents of <5.1. Anti-CSF3 Antibodies - Overview> to <5.6. Origin of Anti-CSF3 Antibodies> above. In one specific example, the artificially engineered protein may be selected from among F(ab), F(ab'), F(ab')2, monospecific F(ab')2, bispecific F(ab')2, single-chain variable fragment (scFv), scFv-Fc, and sdAb. In one specific example, the artificially engineered protein may include any one selected from among F(ab), F(ab'), F(ab')2, monospecific F(ab')2, bispecific F(ab')2, single-chain variable fragment (scFv), scFv-Fc, and sdAb.

[1046] 5.8. Function of anti-CSF3 antibodies or fragments thereof

[1047] 5.8.1. Function to bind to CSF3

[1048] The anti-CSF3 antibody or fragment thereof of the present disclosure may be an antibody or fragment thereof capable of binding to the CSF3 protein. In one specific example, the anti-CSF3 antibody or fragment thereof may be an antibody or fragment thereof that specifically binds to CSF3. In one specific example, the anti-CSF3 antibody may be a monoclonal antibody capable of binding to CSF3. In one specific example, the anti-CSF3 antibody may be a polyclonal antibody capable of binding to CSF3.

[1049] In one specific example, the anti-CSF3 antibody or fragment thereof may bind to the CSF3 protein at a higher level than when binding to other antigens. Specifically, the anti-CSF3 antibody or fragment thereof may bind to the CSF3 protein at a higher level than when binding to ITGA6, AITR, and / or CD58.

[1050] In another specific example, the anti-CSF3 antibody or fragment thereof may bind to the CSF3 protein more strongly than it binds to other antigens. Specifically, the anti-CSF3 antibody or fragment thereof may bind to the CSF3 protein more strongly than it binds to ITGA6, AITR, and / or CD58.

[1051] 5.8.2. Function that inhibits the function of CSF3

[1052] The anti-CSF3 antibody or fragment thereof of the present disclosure can bind to the CSF3 protein, thereby preventing the CSF3 protein from interacting with other molecules, and consequently inhibiting the function of the CSF3 protein. In one specific example, the anti-CSF3 antibody or fragment thereof can inhibit or interfere with the binding of CSF3 to CSF3R. Specifically, the anti-CSF3 antibody or fragment thereof can inhibit or interfere with the binding of CSF3 to CSF3R by binding to CSF3. In another specific example, the anti-CSF3 antibody or fragment thereof can be a neutralizing antibody or fragment thereof against CSF3.

[1053] 5.8.3. Epitope of CSF3

[1054] The specific part of an antigen that allows an antibody to identify it is called an epitope. An epitope can also be referred to as an antigenic determinant.

[1055] The epitope of the human CSF3 protein for the above anti-CSF3 antibody is at least one of KLCATYKLCH (SEQ ID NO: 87); TLDT (SEQ ID NO: 88); SAFQR (SEQ ID NO: 89); and SAFQRR (SEQ ID NO: 90) included in the amino acid sequence constituting the human CSF3 protein (SEQ ID NO: 86).

[1056] In one specific example, the anti-CSF3 antibody can bind to at least one of KLCATYKLCH (SEQ ID NO: 87); TLDT (SEQ ID NO: 88); SAFQR (SEQ ID NO: 89); and SAFQRR (SEQ ID NO: 90) included in the amino acid sequence (SEQ ID NO: 86) constituting the human CSF3 protein. Here, KLCATYKLCH (SEQ ID NO: 87) refers to an amino acid sequence consisting of residues 64 to 73 in the amino acid sequence (SEQ ID NO: 86) constituting the human CSF3 protein. Here, TLDT (SEQ ID NO: 88) refers to an amino acid sequence consisting of residues 132 to 135 in the amino acid sequence (SEQ ID NO: 86) constituting the human CSF3 protein. At this time, SAFQR (SEQ ID NO: 89) refers to an amino acid sequence consisting of residues 172 to 176 in the amino acid sequence (SEQ ID NO: 86) constituting the human CSF3 protein. At this time, SAFQRR (SEQ ID NO: 90) refers to an amino acid sequence consisting of residues 172 to 177 in the amino acid sequence (SEQ ID NO: 86) constituting the human CSF3 protein.

[1057] In one specific example, the anti-CSF3 antibody can bind to ...

Claims

1. Anti-CSF3 (Colony-Stimulating Factor 3) antibody for the treatment of fibrosis. The tissue in which the above fibrosis occurs is one selected from the following: skin, liver, brain, nervous system, heart, bone marrow, gut, joint, eye, kidney, retroperitoneum, mediastinum, and pancreas.

2. In paragraph 1, The above anti-CSF3 antibody is an anti-CSF3 antibody for use in treating fibrosis, which can bind to a human CSF3 protein consisting of an amino acid sequence of SEQ ID NO:

86.

3. In any one of paragraphs 1 and 2, The above anti-CSF3 antibody is an anti-CSF3 antibody for use in treating fibrosis, which induces the breakdown of extracellular matrix accumulated in the tissue where the fibrosis has occurred.

4. In paragraph 3, Anti-CSF3 antibody for the treatment of fibrosis, wherein the extracellular matrix accumulated within the tissue where the fibrosis has occurred is collagen.

5. In any one of paragraphs 1 to 4, The above anti-CSF3 antibody is an anti-CSF3 antibody for use in treating fibrosis, which inactivates myofibroblasts within the tissue where the fibrosis has occurred.

6. In paragraph 5, An anti-CSF3 antibody for use in treating fibrosis, wherein inactivating myofibroblasts within the tissue where said fibrosis has occurred means at least one selected from the following: (a) decomposing the extracellular matrix secreted by the myofibroblasts; (b) inhibiting extracellular matrix secretion of the above myofibroblasts; and (c) Reducing the number of myofibroblasts within the tissue where the fibrosis has occurred.

7. In paragraph 6, The above (c) is an anti-CSF3 antibody for use in treating fibrosis caused by the reversion of the above myofibroblasts into epithelial cells or fibroblasts.

8. In any one of paragraphs 1 to 7, The above anti-CSF3 antibody is an anti-CSF3 antibody for treating fibrosis, which induces the tissue in which the fibrosis has occurred to return to normal tissue.

9. In any one of paragraphs 1 to 8, The above anti-CSF3 antibody is an anti-CSF3 antibody for use in treating fibrosis, which inhibits the activation of TGF-β mediated signaling pathway and non-TGF-β mediated signaling pathway.

10. In paragraph 9, An anti-CSF3 antibody for use in treating fibrosis, wherein activation of the TGF-β mediated signaling pathway induces at least one of epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and extracellular matrix deposition by transforming growth factor beta (TGF-β).

11. In any one of paragraphs 9 to 10, An anti-CSF3 antibody for use in treating fibrosis, wherein activation of the non-TGF-β mediated signaling pathway induces at least one of epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and extracellular matrix deposition through the interaction of CSF3 and CSF3R.

12. A method for preventing or treating fibrosis, comprising: Administering to a subject a therapeutically effective amount of an anti-CSF3 antibody of any one of claims 1 to 11; At this time, the tissue in which the fibrosis occurred is one selected from among skin, liver, brain, nervous system, heart, bone marrow, gut, joint, eye, kidney, retroperitoneum, mediastinum, and pancreas.

13. A pharmaceutical composition for treating fibrosis, comprising: A therapeutically effective amount of an anti-CSF3 antibody of any one of claims 1 to 11; and Pharmaceutically acceptable carrier.

14. In paragraph 13, The pharmaceutical composition for treating fibrosis wherein the pharmaceutically acceptable carrier comprises at least one selected from the following: Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; sweeteners; flavoring agents; syrups; liquid carriers such as fatty oils; sterile aqueous solutions; injectable esters such as propylene glycol; polyethylene glycol; ethyl oleate; suspending agents; emulsions; lyophilized preparations; external preparations; stabilizers; buffers; animal oils; vegetable oils; waxes; paraffin; starches; tragacanth; cellulose derivatives; polyethylene glycols; silicones; bentonites; silica; talc; zinc oxide; and suitable combinations thereof.

15. Use of any one of the anti-CSF3 antibodies of claims 1 to 11 for the manufacture of a fibrosis treatment agent.

16. A method for inhibiting extracellular matrix accumulation and epithelial to mesenchymal transition (EMT) in a target tissue, the method comprising: Administering anti-CSF3 (Colony-Stimulating Factor 3) antibody to the subject; At this time, the above tissue is at risk of extracellular matrix accumulation and EMT. A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the anti-CSF3 antibody can inhibit i) a TGF-β-mediated signaling pathway that induces extracellular matrix accumulation and EMT and ii) a non-TGF-β-mediated signaling pathway that induces extracellular matrix accumulation and EMT.

17. In paragraph 16, A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the anti-CSF3 antibody inhibits the TGF-β mediated signaling pathway and the non-TGF-β mediated signaling pathway, thereby further reducing the expression levels of CSF3, TGF-β, α-SMA, COL1A1, and IL-11 compared to when only the TGF-β mediated signaling pathway is inhibited.

18. In any one of paragraphs 16 to 17, A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the tissue is any one of skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and pancreas.

19. Anti-CSF3R (Colony-Stimulating Factor 3 Receptor) antibody for the treatment of fibrosis. The tissue in which the above fibrosis occurs is one selected from the following: skin, liver, brain, nervous system, heart, bone marrow, gut, joint, eye, kidney, retroperitoneum, mediastinum, and pancreas.

20. In paragraph 19, The above anti-CSF3R antibody is an anti-CSF3R antibody for use in treating fibrosis, which can bind to a human CSF3R protein consisting of an amino acid sequence of SEQ ID NO:

86.

21. In any one of paragraphs 19 to 20, The above anti-CSF3R antibody is an anti-CSF3R antibody for use in treating fibrosis, which induces the breakdown of extracellular matrix accumulated in the tissue where the fibrosis has occurred.

22. In paragraph 21, Anti-CSF3R antibody for the treatment of fibrosis, wherein the extracellular matrix accumulated within the tissue where the fibrosis has occurred is collagen.

23. In any one of paragraphs 19 to 22, The above anti-CSF3R antibody is an anti-CSF3R antibody for use in treating fibrosis, which inactivates myofibroblasts within the tissue where the fibrosis has occurred.

24. In paragraph 23, An anti-CSF3R antibody for use in treating fibrosis, wherein inactivating myofibroblasts within the tissue where said fibrosis has occurred means at least one selected from the following: (a) decomposing the extracellular matrix secreted by the myofibroblasts; (b) inhibiting extracellular matrix secretion of the above myofibroblasts; and (c) Reducing the number of myofibroblasts within the tissue where the fibrosis has occurred.

25. In paragraph 24, The above (c) is an anti-CSF3R antibody for use in treating fibrosis caused by the reversion of the above myofibroblasts into epithelial cells or fibroblasts.

26. In any one of paragraphs 19 to 25, The above anti-CSF3R antibody is an anti-CSF3R antibody for use in treating fibrosis, which induces the tissue in which the fibrosis has occurred to return to normal tissue.

27. In any one of paragraphs 19 to 26, The above anti-CSF3R antibody is an anti-CSF3R antibody for use in treating fibrosis, which inhibits the activation of TGF-β mediated signaling pathway and non-TGF-β mediated signaling pathway.

28. In paragraph 19, An anti-CSF3R antibody for use in treating fibrosis, wherein activation of the TGF-β-mediated signaling pathway induces at least one of epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and extracellular matrix deposition through the interaction of transforming growth factor beta (TGF-β) and its receptor.

29. In any one of paragraphs 27 to 28, An anti-CSF3R antibody for use in treating fibrosis, wherein activation of the non-TGF-β mediated signaling pathway induces at least one of epithelial to mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT), and extracellular matrix deposition through the interaction of CSF3 and CSF3R.

30. A method for preventing or treating fibrosis, comprising: Administering to a subject a therapeutically effective amount of an anti-CSF3R antibody of any one of claims 19 to 29; At this time, the tissue in which the fibrosis occurred is one selected from among skin, liver, brain, nervous system, heart, bone marrow, gut, joint, eye, kidney, retroperitoneum, mediastinum, and pancreas.

31. A pharmaceutical composition for treating fibrosis, comprising: A therapeutically effective amount of an anti-CSF3R antibody of any one of claims 19 to 29; and Pharmaceutically acceptable carrier.

32. In paragraph 31, The pharmaceutical composition for treating fibrosis wherein the pharmaceutically acceptable carrier comprises at least one selected from the following: Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; sweeteners; flavoring agents; syrups; liquid carriers such as fatty oils; sterile aqueous solutions; injectable esters such as propylene glycol; polyethylene glycol; ethyl oleate; suspending agents; emulsions; lyophilized preparations; external preparations; stabilizers; buffers; animal oils; vegetable oils; waxes; paraffin; starches; tragacanth; cellulose derivatives; polyethylene glycols; silicones; bentonites; silica; talc; zinc oxide; and suitable combinations thereof.

33. Use of any one of the anti-CSF3R antibodies of claims 19 to 29 for the manufacture of a fibrosis treatment agent.

34. A method for inhibiting extracellular matrix accumulation and epithelial to mesenchymal transition (EMT) in a target tissue, the method comprising: Administering anti-CSF3R (Colony-Stimulating Factor 3 Receptor) antibodies to the subject; At this time, the above tissue is at risk of extracellular matrix accumulation and EMT. A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the anti-CSF3R antibody can inhibit i) a TGF-β-mediated signaling pathway that induces extracellular matrix accumulation and EMT and ii) a non-TGF-β-mediated signaling pathway that induces extracellular matrix accumulation and EMT.

35. In paragraph 34, A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the anti-CSF3R antibody inhibits the TGF-β mediated signaling pathway and the non-TGF-β mediated signaling pathway, thereby further reducing the expression levels of CSF3, TGF-β, α-SMA, COL1A1, and IL-11 compared to when only the TGF-β mediated signaling pathway is inhibited.

36. In any one of paragraphs 34 to 35, A method for inhibiting extracellular matrix accumulation and epithelial-mesenchymal transition, wherein the tissue is any one of skin, brain, nervous system, heart, bone marrow, liver, gut, joint, eye, lung, kidney, retroperitoneum, mediastinum, and pancreas.

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